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A f i S.&6 - i S 06 90S Sheehy Drive, Bldg. A Horsham, PA 19044 Telephone: (215) 443-3710 Telefax: (215)443-3587 ARGUS RESEARCH Charles River Laboratories Discovery and Development Services Deanna J. Luebker, M.S. 3M Corporate Toxicology 3M Medical Department 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 4 November 2002 S& vJL ^ H A& Y !i/7 Telephone: (651) 737-1374 Telefax: (651)733-1773 RE: Protocol 418-018 - One Generation Reproduction Study of PFOS - Mevalonic Acid/Cholesterol Challenge and NOEL Investigation in Rats Sponsor's Study Number: T-6295.25 Dear Ms. Leubker: Enclosed are the original Good Laboraory Practice Compliance Statement and Certification of Authenticity from Exygen Study 023-069. Please sign these documents and return them to Argus. If you have any questions, please do not hesitate to contact me. RGY :jf Att. Sincerely, Cot Raymond G. York, Ph.D., DABT Associate Director of Research and Study Director ICV=>) mCO KO XDC o m KD C4rD 418-018:PAGE 1-1 TITLE: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS ARGUS RESEARCH.PROTOCOL NUMBER: 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 I. SUMMARY AND CONCLUSION The purposes of this study were to determine whether the co-administration of mevalonic acid or cholesterol could prevent or antagonize the adverse maternal and fetal effects (reduced maternal body weight gain, increased incidence of stillborn, decreased birth weight and decreased pup survival) observed in a previous study with PFOS (Argus Research Protocol 418-008, 3M Study Number T-6295.9) and to better define the noobserved-effect-level (NOEL). A. Methods3 Female Crl:CD(SD) IGS BR VAF/Plus rats were used for the study. The rats were received in two shipments designated as Replicate 1 and 2, respectively. The rats were randomly assigned to 13 dosage groups (Groups 1 through 13), 14 rats per group per replicate for Groups 1, 2, 4 through 7, 12 and 13, 14 rats for Group 3 replicate 1, 15 rats for Group 3, replicate 2, and 10 rats per group per replicate for Groups 8 through 11. The first eight female rats per dosage group in Groups 1 to 7, 12 and 13 of Replicate 1 with a confirmed date of mating were assigned to Caesarean-sectioning on day 21 of gestation (DG 21). The remaining female rats were permitted to naturally deliver litters. The test article was perfluorooctane sulfonic acid potassium salt (PFOS) and the control articles were mevalonic acid and cholesterol. The vehicles were 0.5% Tween 80 and reverse osmosis membrane processed deionized water (RODI H2O). a. Detailed descriptions of all procedures used in the conduct of this study are provided in the appropriate sections of this report and in APPENDIX C (PROTOCOL AND AMENDMENTS). M i The dosage groups and dosing schedule were as follows: 418-018:PAGE 1-2 Number 1 2 3 4 5 6 7 8 9 10 11 12 13 Dosage Groups Identification Dosage Groups and Dosing Schedule Dosing Schedule 2nd dosage 1/2 hour 10 minutes 1st dosage after 1st dosage Vehicle Control RODI H ,0 0.5% Tween 80 Mevalonic Acid Control 1.6 mg/kg PFOS + Mevalonic Acid 2 mg/kg PFOS + Mevalonic Acid Cholesterol Control 1.6 mg/kg PFOS + Cholesterol 2 mg/kg PFOS + Cholesterol 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/kg Cholesterol 500 mg/kg Cholesterol 500 mg/kg Cholesterol 0.5% Tween 80 1.6 mg/kg PFOS 2 mg/kg PFOS 0.5% Tween 80 1.6 mg/kg PFOS 2 mg/kg PFOS 0.4 mg/kg PFOS Not applicable 0.4 mg/kg PFOS 0.8 mg/kg PFOS Not applicable 0.8 mg/kg PFOS 1 mg/kg PFOS Not applicable 1 mg/kg PFOS 1.2 mg/kg PFOS Not applicable 1.2 mg/kg PFOS 1.6 mg/kg PFOS Not applicable 1.6 mg/kg PFOS 2 mg/kg PFOS Not applicable 2 mg/kg PFOS 3rd dosage 5 hr 30 minutes after 2nd dosage RODI HiO 500 mg/kg Mevalonic Acid 500 mg/kg . Mevalonic Acid 500 mg/kg Mevalonic Acid Not applicable Not applicable Not applicable Not applicable Not applicable Not applicable Not applicable Not applicable Not applicable The dosages, concentrations and dosage volumes were as follows: Compound Identification Dosage (mg/kg) Concentration (mg/mL) RODI H20 0 0 0.5% Tween 80 Mevalonic Acid Cholesterol PFOS 0 1000 500 0.4 0 100 100 0.08 PFOS PFOS PFOS 0.8 1 1.2 0.16 0.20 0.24 PFOS 1.6 0.32 PFOS 2 0.40 Mevalonic acid was dosed twice daily. Dosage Volume (mL/kg) 5 5 5* 5 5 5 5 5 5 5 Dosages were adjusted for the most recently recorded body weight and given at approximately the same times each day. 418-018:PAGE 1-3 The rats were given the test article, control article and/or vehicle daily beginning 42 days before cohabitation (maximum of 14 days) and continuing through DG 20 (rats assigned to Caesarean-sectioning), DG 24 (rats assigned to natural delivery that did not deliver a litter) or day 4 of lactation (DL 4, rats that delivered a litter). FI generation pups were not directly given the test article, control article and/or vehicle, but were possibly exposed to the test article or control articles during maternal gestation or via maternal milk during the lactation period. Rats were observed for viability at least twice each day of the study. Observations for clinical signs of effects of the test article and deaths were made daily prior to dosage administration, approximately one hour after the second dosage and at the end of the working day. These observations were also recorded on the day of sacrifice. Body weights were recorded weekly to cohabitation, daily during presumed gestation, on DL 1 (rats assigned to natural delivery) and at sacrifice. Feed consumption values were recorded weekly to cohabitation, on DGs 0, 7, 14, 21 and 25 (if necessary) and DLs 1 and 5 (rats assigned to natural delivery). Mating was evaluated daily during the cohabitation period and confirmed by observation of spermatozoa in a smear of the vaginal contents and/or a copulatory plug observed in situ. The rats assigned to natural delivery were observed for adverse clinical signs during parturition, duration of gestation and pup viability at birth. The rats were also observed for fertility index, gestation index, number of offspring per litter, number of implantation sites, general condition of the dam and litter during the postpartum period and viability index. Maternal behavior was recorded on DLs 1 and 5. Litters were examined after delivery to identify the number and sex of pups, stillboms, live births and gross external alterations. Each litter was evaluated for viability at least twice each day of the 5-day postpartum period. FI generation pups in each litter were counted once daily. Physical signs were recorded each day. Pooled litter weights were recorded on DLs 1 (birth, livebom pups), 2, 3, 4 and 5. Eight rats from dosage groups 1 to 7, 12 and 13 (Replicate 1) were sacrificed on DG 21. Blood and liver samples were collected from these rats. The rats were Caesareansectioned and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number of corpora lutea in each ovary was recorded. The uterus of each rat was excised and examined for pregnancy, number and distribution of implantation sites, live and dead fetuses and early and late resorptions. Placentae were examined for size, color and shape. Blood samples were collected from the rats assigned to Caesarean-sectioning. Following collection of blood samples, the liver of each rat was excised and the liver weight was recorded. A portion of each liver sample (right lateral lobe) was flash frozen and 418-018:PAGE 1-4 maintained frozen. The median liver lobe was frozen. A section from the remaining portion of the liver was fixed in gluteraldehyde. The remaining portion of the liver was frozen. Fetuses were pooled by litter and litter body weights were recorded. Blood samples were collected from each fetus. The liver from each fetus was collected. One fetal liver from each litter was removed and fixed in gluteraldehyde. The remaining fetal livers in each litter were divided into three samples of equal number. Two of the samples were frozen. The remaining samples were flash frozen in liquid nitrogen and maintained frozen. On DL 5 the rats assigned to natural delivery were sacrificed and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Blood, milk, heart and liver samples were collected from maternal rats on DL 5. Following collection of milk and blood samples, the heart and liver of each dam were collected. The liver weight was recorded. A portion of each liver sample (right lateral lobe) was flash frozen and maintained frozen. The median liver lobe was stored frozen. The heart and a section from the remaining portion of the liver were fixed in gluteraldehyde. The remaining portion of the liver was frozen. On DL 5, pups were sacrificed and examined for gross lesions. Necropsy included a single cross-section of the head at the level of the frontal-parietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Blood samples were collected from each pup. For Replicate 1, blood samples were pooled per litter, samples from male and female pups combined. For Replicate 2, blood samples were pooled (two litters per sample within each dosage group). The liver from each pup was collected, pooled (by sex per litter) and the pooled weight recorded. One liver from each litter pool was removed and fixed in gluteraldehyde. The remaining livers in each litter pool were divided into three samples of equal number. Two of the samples were stored frozen. The remaining sample was flash frozen and maintained frozen. The hearts were collected from the first two male and two female pups from each litter (pooled by sex per litter) and were fixed in gluteraldehyde. Thyroids were excised from each pup, pooled (by sex per litter) and retained. Microscopic examination was made of the heart and thyroid from one male and one female FI generation pup from dams of each of the vehicle control (Group 1) and 2 mg/kg/day PFOS groups (Group 13). B. Results No deaths were attributable to the test or control articles. One rat in each of Groups 2 (mevalonic acid control), 3 (1.6 mg/kg PFOS + mevalonic acid) and 5 (cholesterol control) died as the result of intubation accidents or natural causes. All other rats survived to scheduled sacrifice. 000*06 418-018:PAGE 1-5 The numbers of rats with excess salivation, a perioral substance and chromorhinorrhea during gestation were significantly increased in Group 5 (mevalonic acid control) as compared to Group 1 (vehicle control). These increases were attributed to an aversion to the taste of mevalonic acid; the incidence of these observations was comparable among the three mevalonic acid groups (Groups 2, 3 and 4). All other clinical observations during the precohabitation, gestation and lactation periods were considered unrelated to the test or control articles. All necropsy observations were considered unrelated to the test or control articles. Terminal body weights of rats Caesarean-sectioned on DG 21 were significantly reduced in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). The weight of the liver was significantly increased in Group 7. A sa result of the reduced terminal weights in Groups 6 and 7 and the increased liver weight in Group 7, the ratios of the liver weight to the terminal body weight were significantly increased in these two groups. All other terminal body weights, liver weights and relative liver weights of rats Caesareansectioned on DG 21 were unaffected by PFOS or mevalonic acid. Terminal body weights of naturally delivered rats were significantly reduced in Group 13 (2 mg/kg PFOS alone) and in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). The ratios of the liver weights to the terminal body weights were significantly increased in Groups 9, 11 and 13 (0.8, 1.2 and 2 mg/kg PFOS alone, respectively), reflecting slight reductions in terminal body weights and/or slight increases in liver weights in these groups. The liver weight was significantly increased in Group 3 (1.6 mg/kg PFOS + mevalonic acid). The ratios of the liver weight to the terminal body weight were significantly increased in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) and in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). Body weight gains for the entire precohabitation period were significantly reduced in Group 13 (2 mg/kg PFOS alone) and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). During this period, gains were significantly reduced in Groups 6 and 7 on DSs 22 to 29 and 29 to 36; and in Group 13 on DSs 29 to 36. Additionally, body weight gains for Group 12 (1.6 mg/kg PFOS alone) were significantly reduced on DSs 29 to 36 and body weight loss occurred in this group on DSs 36 to 42. As a result of these changes, body weights were significantly reduced in Groups 6 and 7 on DSs 29, 36 and 42. Body weight gains for the entire precohabitation period in Group 5 (cholesterol control) were significantly increased. Premating body weights and body weight gains were unaffected by mevalonic acid and dosages of PFOS as high as 1.2 mg/kg. Body weight gains for the entire gestation period were significantly reduced only in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) and Groups 9, 11, 12 and 13 (0.8, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively) had significantly reduced body weight gains for the first week of the gestation period. There were no significant differences among any of these dosage groups during the second week of gestation. Group 13 had a significant 418-018:PAGE 1-6 increase in body weight gain during the third week of gestation. Body weights were significantly reduced in Group 12 (1.6 mg/kg PFOS alone) on DGs 2 through 16, and in Group 13 (2 mg/kg PFOS alone) on DGs 0 through 19. When PFOS was administered along with mevalonic acid, the effect on body weight was less severe and appeared over a shorter period than PFOS administered alone. Body weights were significantly reduced in Group 3 (1.6 mg/kg PFOS + mevalonic acid) on DGs 4 through 13, and in Group 4 (2 mg/kg PFOS + mevalonic acid) on DGs 7 through 10. The effect on body weight was more severe and appeared over a longer period when PFOS was administered along with cholesterol. Body weights were significantly reduced in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DGs 0 through 21. Gestation body weights were significantly increased in the cholesterol control group (Group 5) on DGs 5, 8 through 17 and 21. Despite this effect of cholesterol alone, body weights in the groups administered PFOS with cholesterol (Groups 6 and 7) had significant reductions in gestation body weights. Body weight gains during the first five days of the lactation period were significantly reduced in Groups 9, 10, 12 and 13 (0.8, 1, 1.6 and 2 mg/kg PFOS alone, respectively). The average body weight on DL 5 was significantly reduced in Group 13. When PFOS was administered along with mevalonic acid, the effect on body weight was less severe than PFOS administered alone. Body weight gains were significantly reduced on DLs 1 to 5 in Group 3, but average body weights on DLs 1 and 5 were comparable among Groups 2, 3 and 4 (mevalonic acid control, 1.6 and 2 mg/kg PFOS + mevalonic acid, respectively). When PFOS was administered along with cholesterol, the effect on body weight was more severe. Body weights on DLs 1 and 5 were significantly reduced in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) and a significant body weight loss occurred on DLs 1 to 5 in Group 7. Absolute and relative feed consumption values were significantly reduced in Group 13 (2 mg/kg PFOS alone) for the entire premating period and within this period on DSs 8 to 15 (relative only), 22 to 29, 29 to 36 and 36 to 42. Absolute and relative feed consumption values were also significantly reduced in Group 12 (1.6 mg/kg PFOS alone) during the last week of the premating period. Absolute feed consumption values were significantly reduced in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) on DSs 22 to 29 and 36 to 42. Relative feed consumption values were significantly reduced in Groups 3 and 4 on DSs 8 to 15, 15 to 22, 22 to 29 and 36 to 42 (Group 4 only). This effect of PFOS and mevalonic acid was reflected in the significantly reduced absolute and relative feed consumption values for the entire premating period in both groups. When PFOS was administered along with cholesterol, the effect on feed consumption values was more severe. Absolute and relative feed consumption values were significantly reduced in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DSs 22 to 29, 29 to 36 and 36 to 42. This effect of PFOS and cholesterol was reflected in the significantly reduced absolute and relative feed consumption values for the entire premating period in both groups. Relative feed consumption values on DSs 8 to 15, 15 to 22, 22 to 29 and 1 to 42 were significantly increased in the mevalonic acid control group (Group 2). Absolute and relative feed consumption values were significantly increased on DSs 29 to 36 and absolute feed M UH 418-018:PAGE 1-7 consumption was significantly increased on DSs 36 to 42 in the cholesterol control group (Group 5). Absolute and relative feed consumption values were significantly reduced during the first week of gestation in Groups 9 through 12 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively). Absolute feed consumption continued to be significantly reduced in Group 13 on DGs 7 to 14, and relative feed consumption was significantly increased in Group 12 on DGs 14 to 21. Absolute feed consumption values for the entire gestation period (DGs 0 to 21) were significantly reduced in Groups 12 and 13 (1.6 and 2 mg/kg PFOS alone, respectively). When PFOS was administered along with mevalonic acid, effects on feed consumption values were comparable to the effects of administration of PFOS alone. Absolute feed consumption values were significantly reduced in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) on DGs 0 to 7, 7 to 14 and 0 to 21. Relative feed consumption values were significantly reduced in Groups 3 and 4 on DGs 0 to 7 and 7 to 14. When PFOS was administered along with cholesterol, effects on feed consumption values were also comparable to the effects of administration of PFOS alone. Absolute feed consumption values were significantly reduced in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DGs 0 to 7, 7 to 14, and 0 to 21. Relative feed consumption values were significantly reduced in Groups 6 and 7 on DGs 0 to 7 and significantly increased in Groups 6 and 7 ( 1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DGs 14 to 21. Absolute feed consumption values during the first five days of the lactation period were significantly reduced in Groups 12 and 13 (1.6 and 2 mg/kg PFOS alone, respectively). Relative feed consumption values for the same period were reduced in Groups 9 through 13, reaching statistical significance in Groups 9, 10, 11 and 12 (0.8, 1, 1.2 and 1.6 mg/kg PFOS alone, respectively) values. When PFOS was administered along with mevalonic acid, effects on feed consumption values were also comparable to the effects of administration of PFOS alone. Absolute and relative feed consumption values were significantly reduced in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) on DLs 1 to 5. When PFOS was administered along with cholesterol, effects on feed consumption values were comparable to the effects of administration of PFOS alone. Absolute and relative feed consumption values were significantly reduced in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DLs 1 to 5. The number of days in cohabitation, the fertility and pregnancy indices and the number of rats with confirmed mating dates during the first and second week of cohabitation were comparable in the thirteen dosage groups. Caesarean-sectioning observations on DG 21 were based on eight pregnant dams in each of Groups 1 through 7, 12 and 13. No Caesarean-sectioning or litter parameters were affected by dosages up to 2 mg/kg PFOS alone or administered with either cholesterol or mevalonic acid. Totals of 17 to 20 rats were pregnant and delivered litters in each of the 13 dosage groups. The duration of gestation was significantly reduced in Groups 9 through 13 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). The number of dams with all pups dying on DLs 1 to 5 was 000009 418-018:PAGE 1-8 increased in Groups 13, 3, 4, 6 and 7; the increase was significant in Groups 13, 4 and 7. The pup viability index was significantly reduced in Groups 12 and 13 (1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). The numbers of pups that died or were presumed cannibalized on DL 1 and DLs 2 to 5 was significantly increased in Groups 12, 13, 4, 6 and 7. The numbers of surviving pups per litter were significantly reduced on DLs 2, 3, 4 and 5 in Groups 12, 13, 3, 4, 6 and 7. A dosage-dependent pattern of reduced pup body weight was evident on DLs 1, 2, 3, 4, 5 and/or 1 to 5 in each group administered the test article. Groups 11, 12 and 13 (1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) had significantly reduced pup body weights on most weighing days. When average pup weight was normalized for litter size, Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) and Groups 8, 9, 10, 11, 12 and 13 (0.4, 0.8, 1, 1.2, 1.6 and 2 mg/kg/day PFOS, respectively) were also significantly reduced from Group 2 (mevalonic acid control, Group 5 (cholesterol control) and Group 1 (Vehicle Control), respectively, over the same days of lactation. Adverse clinical and necropsy observations associated with reduced pup viability and potential reduction in maternal care occurred in Groups 11,12 and 13 (1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol). Increased numbers of litters with pups that were cold to touch occurred in Groups 11, 12, 13, 4, 6 and 7. The number of litters with pups not nursing was increased in Groups 12, 13, 3, 4, 6 and 7; the increase was significant in Group 7. Necropsy observations in pups that were stillborn or found dead included increases in the numbers of pups with no milk in the stomach in Groups 9, 10, 11, 12 and 13 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively). The incidence of this observation was significantly increased in Group 6. All other clinical and necropsy observations in the FI generation pups were considered unrelated to the test article. Clinical chemistry parameters were generally unaffected in the DG 21 dams that were treated with or without cholesterol and 1.6 or 2 mg/kg PFOS. Total and free T3 and T4 in dams administered 1.6 or 2 mg/kg PFOS with or without mevalonic acid or cholesterol were significantly reduced. The LDL was significantly increased in the dams that were administered mevalonic acid and PFOS. Liver cholesterol levels were significantly reduced in DG 21 dams administered 1.6 or 2 mg/kg PFOS and in DG 21 dams coadministered mevalonic acid and 1.6 or 2 mg/kg PFOS. Liver triglyceride levels were significantly reduced in DG 21 dams coadministered mevalonic acid or cholesterol and 1.6 or 2 mg/kg PFOS. Liver LDL and HDL levels were increased or significantly increased in dams coadministered cholesterol or mevalonic acid and 1.6 or 2 mg/kg PFOS. 000010 418-018:PAGE 1-9 Cholesterol and LDL were significantly increased in the fetuses from dams that were treated with 1.6 or 2 mg/kg PFOS. Free T4 was significantly reduced in the fetuses from dams that were treated with or without cholesterol or mevalonic acid and 1.6 or 2 mg/kg PFOS. Cholesterol and LDL were significantly increased and triglycerides were significantly decreased in the fetuses from dams that were treated with mevalonic acid and 1.6 and/or 2 mg/kg PFOS. The clinical chemistry parameters of cholesterol, LDL and HDL were significantly increased in the fetuses from dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS. Liver cholesterol and triglyceride levels were increased or significantly increased in DG 21 fetuses from dams coadministered mevalonic acid or cholesterol and 1.6 or 2 mg/kg PFOS. Liver HDL levels were significantly increased in DG 21 fetuses from dams administered 1.6 or 2 mg/kg PFOS alone or coadministered mevalonic acid and 1.6 or 2 mg/kg PFOS. Serum cholesterol was significantly decreased in the lactating dams that were treated with 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS. The glucose was significantly increased in the DL 5 dams that were treated with 2 mg/kg PFOS alone and triglycerides were significantly decreased in lactating dams treated with 1.6 or 2 mg/kg PFOS alone. Total and free T4 levels in lactating dams administered 0.4 (free T4 only), 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced and total and free T3 levels in lactating dams administered 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced. Serum cholesterol was significantly decreased in the lactating dams that were treated with mevalonic acid and 1.6 or 2 mg/kg PFOS and cholesterol in the milk was significantly increased in the lactating dams that were treated with mevalonic acid and 2 mg/kg PFOS. Total and free T4 were significantly reduced in the lactating dams that were treated with mevalonic acid or cholesterol and 1.6 or 2 mg/kg PFOS. Cholesterol in the blood and triglycerides were significantly decreased in lactating dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS and glucose was significantly increased in lactating dams that were treated with cholesterol and 2 mg/kg PFOS. Liver triglyceride levels were increased or significantly increased in DL 5 dams administered 1.6 or 2 mg/kg PFOS alone and in DL 5 dams coadministered mevalonic acid or cholesterol and 1.6 or 2 mg/kg PFOS. Clinical chemistry parameters were not significantly different in the DL 5 pups from lactating dams that were treated with 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS. Free T4 levels in the FI generation pups from lactating dams administered 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced. Free T3levels in pups from lactating dams administered 0.4, 0.8 or 1 mg/kg PFOS were significantly reduced, however, levels were significantly increased, in pups from lactating dams administered 1.6 mg/kg PFOS. Total T4 levels in pups from lactating dams administered 0.4, 0.8 or 1 mg/kg PFOS were significantly reduced. TSH was significantly increased at 1 mg/kg PFOS. Liver triglyceride levels were reduced or significantly reduced in DL 5 male and female pups from dams administered 1, 1.2, 1.6 and 2 mg/kg PFOS alone and in DL 5 male pups from dams coadministered cholesterol and 1.6 or 2 mg/kg PFOS. Cholesterol, LDL and triglycerides in the blood were significantly increased in pups from lactating dams that were treated with mevalonic acid and 1.6 mg/kg PFOS. Total T4were significantly decreased in pups from lactating dams that were treated with mevalonic acid and 1.6 mg/kg PFOS. LDL and HDL were significantly increased in the pups from MMU 418-018:PAGE MO lactating dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS and triglycerides were significantly decreased in the pups from dams that were treated with cholesterol and 2 mg/kg PFOS. Total T3 and free T4 were significantly decreased in pups from lactating dams that were treated with cholesterol and 1.6 and/or 2 mg/kg PFOS. No microscopic changes were seen in the heart or thyroid of the male and female pups whose dam was exposed to 2 mg/kg/day PFOS (Group 13). C. Conclusion On the basis of these data, it does not appear that coadministration of mevalonic acid or cholesterol can prevent or antagonize the adverse maternal or fetal effects observed in a previous study with PFOS (Argus Research protocol 418-008; 3M study number T-6295.9). The maternal no-observable-effect-level (NOEL) for this study is 0.4 mg/kg PFOS (the 0.8 mg/kg and higher dosages of PFOS caused an increased liver weight to terminal body weight ratio, decreased body weight gain during gestation and lactation and decreased absolute and relative feed consumption during gestation and lactation on days). The reproductive NOEL in the dams is also 0.4 mg/kg (the 0.8 mg/kg dosage decreased the duration of gestation). The NOEL for viability and growth in the offspring is less than 0.4 mg/kg (dosages of 0.4 mg/kg and higher caused decreased pup body weights when normalized for litter size). Georgle&/E. Dearlove, Ph.D., DABT Associate Director of Research Date Associate Director of Research and Study Director 418-018:PAGE E-l II. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study A .l. Sponsor 3M Corporate Toxicology, 3M Center, Building 220-2E-02, St. Paul, Minnesota 55144-1000 A.2. Testing Facility Argus Research, 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 A.3, Study Number 418-018 A.4. Sponsor's Study Number T-6295.25 A.5. Purpose of the Study The purposes of this study were: 1) to determine whether or not co-administration of mevalonic acid or cholesterol can prevent or antagonize the adverse maternal and fetal effects (reduced maternal body weight gain, increased percent stillborn, decreased birth weight and decreased pup survival) observed in a previous study (Argus Research Protocol 418-008, 3M study number T-6295.9); and 2) to better define the no observed effect level (NOEL). A.6. Study Design The requirements of the U.S. Food and Drug Administration (FDA)(1)were used as the basis for the study design. A .l. Regulatory Compliance The requirements of the U.S. Food and Drug Administration (FDA)(1) were used as the basis for the study design. The study was conducted in compliance with Good Laboratory Practice (GLP) regulations of the U.S. Food and Drug Administration (FDA) , the Japanese Ministry of Health and Welfare (MHW)(3) and the European Economic Community (EEC)(4). There were no deviations from the GLP regulations that affected the quality or integrity of the study. Quality Assurance Unit findings derived from the inspections during the conduct of this study are documented and have been provided to the Study Director and the Testing Facility Management. 418-018:PAGE H-2 A.8. Ownership of the Study The Sponsor owns the study. All raw data, analyses, reports and preserved tissues are the property of the Sponsor. A.9. Study Monitor Deanna J. Luebker, M.S. A.10, Alternate Study Monitor John Butenhoff, Ph.D., DABT, CIH A .ll. Study Director Raymond G. York, Ph.D., DABT A.12. Technical Performance John F. Barnett, B.S. (Director of Laboratory Operations) Todd J. Killino, B.S. (Research Associate) Alaine L. Casey, A.S. (Laboratory Technician) Brian K. Kelsch (Necropsy Laboratory Technician) Christopher K. Ruppert, B.S. (Formulation Laboratory Technician) A.13. Report Preparation Raymond G. York, Ph.D., DABT Jo Ann Frazee, M.S. (Study Coordinator) Pamela J. Shufelt, B.S. (Data Management Team Leader) Cristina Petrescu (Report Administrator) A.14. Report Review Valerie A. Sharper, M.S. (Director of Study Management) A.15. Date Protocol Signed 21 August 2000 HW i 418-018:PAGE II-3 A.16. Dates of Technical Performance A.16.a. Replicate 1 Rat Arrival Dosage Period Rats Assigned to Caesarean-Sectioning (42 days before cohabitation and continuing through DGa 20) Rats Assigned to Natural Delivery [42 days before cohabitation and continuing through DG 24 (rats that did not deliver a litter) or DLb4 (rats that delivered a litter)] Cohabitation Period Male 1 (7 days maximum) Male 2 (7 days maximum) Scheduled Sacrifice (DG 21 Caesarean-sectioning) DG 25 Sacrifice No Confirmed Date of Mating Sacrifice Delivery Period (DL 1) Scheduled Sacrifice (DL 5) 22 AUG 00 29 AUG 00 - 02 NOV 00 29 AUG 00- 18 NOV 00 09 OCT 00 PM - 16 OCT 00 AM 16 OCT 00 PM - 23 OCT 00 AM 31 OCT 00-03 NOV 00 05 NOV 00 - 09 NOV 00 17 NOV 00 31 OCT 00- 15 NOV 00 04 NOV 00-19 NOV 00 A.16.b. Replicate 2 Rat Arrival Dosage Period Rats Assigned to Natural Delivery [42 days before cohabitation and continuing through DG 24 (rats that did not deliver a litter) or DL 4 (rats that delivered a litter)] Cohabitation Period Male 1 (7 days maximum) Male 2 (6 days maximum) DG 25 Sacrifice Delivery Period (DL 1) Scheduled Sacrifice (DL 5) 05 SEP 00 14 SEP 00 - 02 DEC 00 25 OCT 00 PM - 01 NOV 00 AM 01 NOV 00 PM - 07 NOV 00 AM 20 NOV 00 - 23 NOV 00 16 NOV 00 - 29 NOV 00 20 NOV 00 - 03 DEC 00 a. DG is an abbreviation for day of (presumed) gestation. b. DL is an abbreviation for day of lactation/postpartum. 418-018:PAGE II-4 A.17. Records Maintained The original report, raw data and reserve samples of each lot of the control article and vehicle components are retained in the archives of Argus Research. Any preserved tissues are retained in the archives of the Testing Facility for one year after mailing the draft final report, after which time the Sponsor will decide their final disposition. All unused prepared formulations were discarded at the Testing Facility. Unused bulk test article was returned to the Sponsor. B. Test Article Information B.l. Description Perfluorooctane Sulfonic Acid Potassium Salt (PFOS) - light-colored powder B.2. Lot Number 217 B.3. Date Received and Storage Conditions The test article was received on 25 August 2000, and stored at room temperature. B.4. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask and safety goggles or safety glasses and a face-shield) were taken when handling the test article. B.5. Analysis of Activity Information regarding the identity, composition, strength, and purity of the test article is on file with the Sponsor. The purity is 98.9%. A Certificate of Analysis is available in APPENDIX E. C. Control Articles Information C.l. Descriptions Mevalonic Acid - white with a yellow cast, semisolid Cholesterol - white powder C.2. Lot Numbers Mevalonic Acid - 070K26603 and 080K2618 Cholesterol - 119H0218 000*10 418-018:PAGE II-5 C.3. Dates Received and Storage Conditions The mevalonic acid was received from Sigma Chemical Company, St. Louis, Missouri, on 1 August 2000 (lot 070K26603) and 21 September 2000 (lot 080K2618), and stored frozen. The cholesterol was received from Sigma Chemical Company, St. Louis, Missouri, on 20 July 2000, and stored at room temperature. C.4. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask and safety goggles or safety glasses and a face-shield) were taken when handling the control articles. C.5. Analysis of Purity Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the control articles that would have interfered with the results of this study. The purity of the mevalonic acid is approximately 97%. The purity of the cholesterol is 95%. The expiration dates for the mevalonic acid are August 2004 (lot 070K26603) and September 2004 (lot 080K2618). The expiration date for the cholesterol is July 2004. D. Vehicle Information D .l. Description 0.5% Tween 80 prepared using Tween 80, a yellow liquid, and reverse osmosis membrane processed deionized water (RODI H2O). Reverse osmosis membrane processed deionized water (RODI H2O). D.2. Lot Number N32477 D.3. Dates Received and Storage Conditions The Tween 80 was received from J.T. Baker, Phillipsburg, New Jersey, on 7 March 2000, and stored at room temperature. Reverse osmosis membrane processed deionized water is available from a continuous source at the Testing Facility and is maintained at room temperature. D.4. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask and safety goggles or safety glasses and a face-shield) were taken when handling the vehicles. U U 1T 418-018:PAGE II-6 D.5. Analysis of Purity Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the vehicles that would have interfered with the results of this study. The expiration date for the Tween 80 is March 2004. E. Test Article Preparation and Storage Conditions PFOS suspensions and the 0.5% Tween 80 were prepared weekly at the Testing Facility. Mevalonic acid was prepared twice daily in reverse osmosis membrane processed deionized water. Cholesterol was prepared as a suspension once daily in 0.5% Tween 80. The prepared vehicle (0.5% Tween 80) was stored at room temperature. The prepared test article formulations were stored frozen (-20C). E.l. Sample Information Sample Type Date Storage/Shipping Size Retained Conditions Shipped To Date Shipped Homogeneity3 Concentration0 Control Articles Reserve Mevalonic Acid Lot 070K26603 Lot 080K2618 Cholesterol Vehicle Component Reserves Tween 80 RODI H20 5 mL 5 mL lg 1g 1g 5 mL 5 mL 28 AUG 00 27 SEP 00b 18 SEP 00 01 NOV 00 15 NOV 00 Frozen (-20C) Frozen (-20C) Sponsor Sponsor 01 SEP 00 30 OCT 00 01 SEP 00 01 SEP 00 01 SEP 00 Frozen Frozen Room temperature Testing Facility Archives Room temperature Room temperature Testing Facility Archives 29 AUG 00 27 SEP 00 18 SEP 00 01 NOV 00 15 NOV 00 13 SEP 00 08 NOV 00 13 SEP 00 13 SEP 00 13 SEP 00 a. A syringe was used to withdraw samples from the top, middle and bottom of the highest PFOS concentration on the first day of preparation. Each sample was divided into two aliquots and placed into glass containers, one of 2 mL and one of 3 mL. One aliquot (2 mL) was shipped for analysis; the other aliquot (3 mL) was retained at the Testing Facility as a backup sample. b. A larger preparation size was required for dosing; therefore, according to the Standard Operating Procedures of the Testing Facility, if preparation is 30% larger or smaller, homogeneity must be reanalyzed. c. A syringe was used to withdraw samples from each PFOS concentration once during each of the following periods of study: premating, gestation and postnatal. Each sample was divided into two aliquots, one of 2 mL and one of 3 mL, and placed into glass containers. One aliquot (2 mL) was shipped for analysis; the other aliquot (3 mL) was retained at the Testing Facility as a backup sample. 00001 418-018:PAGE n -7 E.2. Analytical Results Samples were sent to Exygen Research, State College, Pennsylvania, for analysis. PFOS levels in the Day 0 and 10 stability and concentration samples bracketing the range of concentrations and conditions of this study were determined. Analysis of samples on 14 February 2002 to 15 February 2002 showed an average percent recovery standard deviation for PFOS in the dosing solutions as 93% 7% and ranged from 83% to 105%, when compared to the assigned concentration, which was not considered to be significant. Results of the stability and concentration analyses are available in APPENDIX F. F. Test System F.l. Species Rat F.2. Strain Crl:CD(SD) IGS BR VAF/Plus F.3. Supplier (Source Charles River Laboratories, Inc., Raleigh, North Carolina F.4. Sex Female (Note: Male rats were used only for the purpose of breeding and are not considered part of the Test System). F.5. Rationale for Test System The Crl:CD(SD) IGS BR VAF/Plus rat was selected as the Test System because: 1) this strain of rat has been demonstrated to be sensitive to reproductive and developmental toxins and has been widely used throughout industry for reproductive and developmental toxicity evaluations; 2) historical data and experience exist at the Testing Facility(5'7); 3) the test article is pharmacologically active in the species and strain; and 4) this strain has been used in previous reproduction studies of PFOS. 000019 418-018:PAGE II-8 F.6. Test System Data Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) the Day after Arrival2 Weight (g) at Study Assignment Replicate 1 180 27 JUN 00 57 days 167 - 204 196 - 224 Replicate 2 179 10 JUL 00 58 days 170 - 228 199 - 247 F.7. Breeder Male Rat Data Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) the Day after Arrival Weight (g) at Cohabitation - Replicate 1 Weight (g) at Cohabitation - Replicate 2 Shipment 1 150 29 FEB 00 71 days 245 - 339 473 - 767 452 - 773 Shipment 2 150 20 MAR 00 72 days 286 - 344 537 - 748 593 - 791 F.8. Method of Randomization Female rats were received in two shipments, separated by two weeks. The first shipment was designated as Replicate 1 and the second shipment was designated as Replicate 2. Upon arrival, the male and female rats were assigned to individual housing on the basis of computer-generated random units. After acclimation, female rats were selected for study on the basis of physical appearance and body weights recorded during acclimation. The rats were assigned to 13 dosage groups (Groups 1 through 13). There were 14 rats per group per replicate for Groups 1, 2, 4 through 7, 12 and 13, 14 rats for Group 3, replicate 1, 15 rats for Group 3, replicate 2, and 10 rats per group per replicate for Groups 8 through 11. Computer-generated (weight-ordered) randomization procedures were used to assign rats to groups. The first eight female rats per dosage group in Groups 1 to 7, 12 and 13 of Replicate 1 with a confirmed date of mating were assigned to Caesarean-sectioning on DG 21. The remaining female rats (including those with no confirmed mating date) were permitted to naturally deliver litters. F.9. System of Identification Male rats were given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats were assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study. Each rat was individually identified with a Monel self-piercing ear tag (Gey Band and Tag Co., Inc., No. MSPT 20101) inscribed with the rat's designated a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), item 1. 00*929 418-018-.PAGE II-9 unique permanent number. Cage tags were marked with the study number, permanent rat number, sex, test article identification and dosage level. Pups were not individually identified during lactation, all parameters were evaluated in terms of the litter. G. Husbandry G.l. Research Facility Registration USDA Registration No. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 et seq. G.2. Study Rooms The study rooms were maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum of ten changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters. Room temperature and humidity were monitored constantly throughout the study. Room temperature was targeted at 64F to 79F (18C to 26C); relative humidity was targeted at 30% to 70%a. G.3. Housing Rats were individually housed in stainless steel, wire-bottomed cages, except during cohabitation and postpartum periods. During cohabitation, each pair of rats was housed in the male rat's cage. Beginning no later than DG 20, Fo generation female rats assigned to natural delivery were individually housed in nesting boxes. Each dam and delivered litter was housed in a common nesting box during the postpartum period. All cage sizes and housing conditions were in compliance with the Guide for the Care and Use of Laboratory Animals(8). G.4. Lighting An automatically-controlled fluorescent light cycle was maintained at 12-hours light: 12-hours dark, with each dark period beginning at 1900 hours EST. G.5. Sanitization Cage pan liners were changed approximately three times each week. Cages were changed approximately every other week. Bedding was changed as often as necessary to keep the rats clean and dry. a. See APPENDIX G (TEMPERATURE AND RELATIVE HUMIDITY REPORTS). 0 0 1Ti>'4i 418-018-.PAGE H-10 G.6. Feed Rats were given ad libitum access to Certified Rodent Diet #5002 (PMI Nutrition International, St. Louis, Missouri) in individual feeders. G.7. Feed Analysis Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration for certified feed or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the feed that would have interfered with the results of this study. G.8. Water Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad libitum from an automatic watering system and/or individual water bottles attached to the cages. Chlorine was added to the processed water as a bacteriostat. G.9. Water Analysis The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, Pennsylvania) and monthly for possible bacterial contamination (Analytical Laboratories, Inc., Chalfont, Pennsylvania). Copies of the results of the water analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the water that would have interfered with the results of this study. G.10. Nesting Material Bed-o'cobs bedding (The Andersons Industrial Products Group, Maumee, Ohio) was used as the nesting material. G .ll. Bedding Analysis Analyses for possible contamination are conducted semi-annually. Copies of the results of the bedding analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the bedding that would have interfered with the results of this study. 000*22 H. Methods H .l. Dosage Administration The dosage groups and dosing schedule were as follows2. 418-018:PAGE II-11 Dosage Groups Number 1 Identification Vehicle Control Dosage Groups and Dosing Schedule 1st dosage Dosing Schedule 2nd dosage 1/2 hour 10 minutes after 1st dosage RODI H20 0.5% Tween 80 2 Mevalonic Acid 500 mg/kg Control Mevalonic Acid 3 1.6 mg/kg PFOS + 500 mg/kg Mevalonic Acid Mevalonic Acid 4 2 mg/kg PFOS + 500 mg/kg Mevalonic Acid Mevalonic Acid 5 Cholesterol Control 500 mg/kg Cholesterol 6 1.6 mg/kg PFOS + Cholesterol 500 mg/kg Cholesterol 7 2 mg/kg PFOS + Cholesterol 500 mg/kg Cholesterol 8 0.4 mg/kg PFOS Not Applicable 9 0.8 mg/kg PFOS Not Applicable 0.5% Tween 80 1.6 mg/kg PFOS 2 mg/kg PFOS 0.5% Tween 80 1.6 mg/kg PFOS 2 mg/kg PFOS 0.4 mg/kg PFOS 0.8 mg/kg PFOS 10 l mg/kg PFOS Not Applicable 11 1.2 mg/kg PFOS Not Applicable 12 1.6 mg/kg PFOS Not Applicable 1 mg/kg PFOS 1.2 mg/kg PFOS 1.6 mg/kg PFOS 13 2 mg/kg PFOS Not Applicable 2 mg/kg PFOS 3rd dosage 5 hours 30 minutes after 2nd dosage RODI H20 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid Not Applicable Not Applicable Not Applicable Not Applicable Not Applicable Not Applicable Not Applicable Not Applicable Not Applicable a. See APPENDIX D, items 2 through 4. 418-018:PAGE n-12 The dosages, concentrations and dosage volumes were as follows2: Compound Identification Dosage (mg/kg/day) Concentration (mg/mL) Dosage Volume (mL/kg) RODI H20 0 0 5 0.5% Tween 80 0 0 5 Mevalonic Acid 1000 100 5* Cholesterol 500 100 5 PFOS 0.4 0.08 5 PFOS 0.8 0.16 5 PFOS 1 0.20 5 PFOS 1.2 0.24 5 PFOS 1.6 0.32 5 PFOS 2 0.40 5 The test and control articles were considered 100% pure for the purpose of dosage calculations. * Mevalonic acid was dosed twice daily. The rats were assigned to groups as follows: Dosage Group 1 2 3 4 5 Identification Vehicle Control Mevalonic Acid Control 1.6 mg/kg PFOS + Mevalonic Acid 2 mg/kg PFOS + Mevalonic Acid Cholesterol Control 6 1.6 mg/kg PFOS + Cholesterol 7 2 mg/kg PFOS + Cholesterol 8 0.4 mg/kg PFOS 9 0.8 mg/kg PFOS 10 1 mg/kg PFOS 11 1.2 mg/kg PFOS 12 1.6 mg/kg PFOS 13 2 mg/kg PFOS Number of Rats 28 28 29 28 28 28 28 20 20 20 20 28 28 Assigned Rat Numbers Replicate 1 Replicate 1 Caesarean-sectioning Natural Delivery 10901 - 10905, 10907, 10906,10909,10911, 10908, 10910,10912 10913, 10914 10915- 10920, 10925, 10921 - 10924, 10926, 10928 10927 10930-10935, 10937, 10929, 10936, 10938, 10940 10939, 10941, 10942 10945-10947, 10949- 10943, 10944, 10948, 10951, 10953, 10954 10952, 10955, 10956 10958 - 10965 10957, 10966 - 10970 10972, 10974- 10976, 10978, 10980- 10982 10971, 10973, 10977, 10979, 10983, 10984 10985, 10987 - 10993 10986, 10994- 10998 Not applicable 10999- 11008 Not applicable 11009- 11018 Not applicable 11019- 11028 Not applicable 11029- 11038 11040, 11039, 11044- 11047, 11041 - 11043,11048, 11050- 11052 11049 11053, 11054, 11056, 11058, 11060, 11062, 11063, 11066 11055, 11057, 11059, 11061, 11064,11065 Replicate 2 Natural Delivery 11501 - 11514 11515- 11528 11529- 11543 11544- 11557 11558- 11571 11572- 11585 11586- 11599 11600-11609 11610-11619 11620- 11629 11630-11639 11640- 11653 11654- 11667 a. See APPENDIX D, items 5 through 7. 00024 418-018:PAGE 11-13 H.2. Rationale for Dosage Selection Dosages were selected by the Sponsor on the basis of previous studies conducted with the test article. H.3. Route and Rationale for Route of Administration The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered; 2) it is one of the possible routes of human exposure; and 3) it is the route used in previous reproduction studies on PFOS. H.4. Method and Frequency of Administration Dosages were adjusted for the most recently recorded body weight and given at approximately the same times each day. Dosages were administered according to the previous table. The mevalonic acid dosing needle was wiped clean before administration for each rat. Female rats were given the test article, control article and/or vehicle daily beginning 42 days before cohabitation (maximum of 14 days) and continuing through DG 20 (rats assigned to Caesarean-sectioning), DG 24 (rats assigned to natural delivery that did not deliver a litter) or DL 4 (rats that delivered a litter). Rats in the process of delivering were not given test/control article; such rats may not have received any or all of their daily dosage(s) on the day of parturition. FI generation pups were not directly given the test article, control article and/or vehicle, but were possibly exposed to the test article during maternal gestation (in utero exposure) or via maternal milk during the lactation period. H.5. Method of Study Performance H.5.a. Fo Generation Rats Within each dosage group, consecutive order was used to assign rats to cohabitation, one male rat per female rat. The cohabitation period consisted of a maximum of 14 days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ were considered to be at DG 0 and assigned to individual housing. Female rats not mated within the first seven days of cohabitation were assigned alternate male rats that had mated and remained in cohabitation for a maximum of seven additional days. Rats were observed for viability at least twice each day of the study. Rats were examined for clinical observations and general appearance at least once during the acclimation period3. Observations for clinical signs of effects of the test article and deaths were made a. See APPENDDC D, items 8 and 9. 418-018:PAGE 11-14 daily prior to dosage administration, approximately one hour after the second dosage and at the end of the working day (approximately one hour after the third dosage)2. These observations were also recorded on the day of sacrifice. Body weights were recorded at least once during the acclimation period, weekly to cohabitation, daily during presumed gestation, on DL 1 (rats assigned to natural delivery) and at sacrifice2. Feed consumption values were recorded weekly to cohabitation, on DGs 0, 7, 14, 21 and 25 (if necessary) and DLs 1 and 5 (rats assigned to natural delivery)6. During cohabitation when two rats occupied the same cage with one feed jar, replenishment of feed jars was documented, but individual values were not recorded or tabulated. Mating was evaluated daily during the cohabitation period and confirmed by observation of spermatozoa in a smear of the vaginal contents and/or a copulatory plug observed in situ. The rats assigned to natural delivery were observed for adverse clinical signs during parturition, duration of gestation (DG 0 to the day the first pup was observed) and pup viability at birth. The rats were also observed for fertility index (percentage of matings that result in pregnancy), gestation index (percentage of pregnancies that result in birth of live litters), number of offspring per litter (live and dead pups), number of implantation sites, general condition of the dam and litter during the postpartum period and viability index (percentage of pups bom that survived five days). Maternal behavior was recorded on DLs 1 and 5C. Deviations from expected maternal behavior were recorded, when and if observed, on all other days of the postpartum period. H.5.b. FI Generation Pups Litters were examined after delivery to identify the number and sex of pups, stillboms, live births and gross external alterations. Day 1 of lactation (postpartum) was defined as the day of birth and was also the first day on which all pups in a litter were weighed (pup body weights were recorded after all pups in a litter were delivered and groomed by the dam). Each litter was evaluated for viability at least twice each day of the 5-day postpartum period. Dead pups observed at these times were removed from the nesting box. FI generation pups in each litter were counted once daily. Physical signs (including variations from expected lactation behavior and gross external alterations) were recorded each day. Pooled litter weights were recorded on DLs 1 (birth, livebom pups), 2, 3, 4 and 5. a. See APPENDIX D, items 8 and 9. b. See APPENDIX D, item 10. c. See APPENDIX D, item 11. 90W 26 418-018:PAGE 11-15 H.6. Gross Necropsy3 H.6.a. Fo Generation Rats Assigned to Caesarean-Sectioning Eight rats from Groups 1 to 7, 12 and 13 (Replicate 1) were sacrificed by carbon dioxide asphyxiation on DG 21. Blood and liver samples were collected from these ratsb. The rats were Caesarean-sectioned and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Gross lesions were retained in neutral buffered 10% formalin for possible future evaluation. Representative photographs of gross lesions are available in the raw data. Uteri of apparently nonpregnant rats were stained with 10% ammonium sulfide to confirm the absence of implantation sites(9) and retained in neutral buffered 10% formalin for possible future evaluation. The number of corpora ltea in each ovary was recorded. The uterus of each rat was excised and examined for pregnancy, number and distribution of implantation sites, live and dead fetuses and early and late resorptions. An early resorption was defined as one in which organogenesis was not grossly evident. A late resorption was defined as one in which the occurrence of organogenesis was grossly evident. A live fetus was defined as a term fetus that responded to stimuli. Nonresponding fetuses were considered to be dead. Dead fetuses and late resorptions were differentiated by the degree of autolysis present; marked to extreme autolysis indicated that the fetus was a late resorption. Placentae were examined for size, color and shape. Blood samples (at least 4 mL each) were collected from the rats assigned to Caesareansectioning via the inferior vena cava and separated into two approximately equal aliquots. The first aliquot was placed into EDTA-coated tubes and the second aliquot was transferred into serum separator tubes. The samples were spun in a centrifuge and the resulting serum (divided into two aliquots) and plasma (one aliquot) were transferred into labeled polypropylene tubes. All samples were immediately frozen on dry ice and maintained frozen (<-20 C). Following collection of blood samples, the liver of each rat was excised and the liver weight was recorded0. A portion of each liver sample (right lateral lobe) was flash frozen in liquid nitrogen and maintained frozen (<-70C). The median liver lobe was stored frozen (<-20C). A section (approximately 0.3 to 0.5 cm wide) from the remaining portion of the liver was fixed in gluteraldehyde for possible electron microscopy. The remaining portion of the liver was stored frozen (<-20C). Fetuses were pooled by litter and litter weights were recorded*1. a. A table of random units was used to select one control group rat assigned to Caesarean-sectioning from which all tissues examined at necropsy were retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions. b. See APPENDIX D, item 12. c. See APPENDIX D, item 13 d. See APPENDIX D, item 14. 000927 418-018-.PAGEII-16 Caps and labeled tubes were weighed (combined weight, to the nearest .001 gram) before (empty) and after retention of pooled fetus samples for subsequent use in pharmacokinetic analyses. Sample tubes were labeled with the study number, litter identification, date of collection, study day, sample identification and collection timepoint. Blood samples were collected from each fetus via decapitation. Blood from approximately one-half of the fetuses in each litter was placed into EDTA-coated tubes and the remaining fetal blood was transferred into serum separator tubes. The samples were spun in a centrifuge and the resulting serum (divided into two aliquots) and plasma (one aliquot) were transferred into labeled polypropylene tubes. All samples were immediately frozen on dry ice and maintained frozen (<-20C). The liver from each fetus was collected. One fetal liver from each litter was removed and fixed in gluteraldehyde for possible electron microscopy. The remaining fetal livers in each litter were divided into three samples of equal number (when possible). Two of the samples were stored frozen (<-20C). The remaining samples were flash frozen in liquid nitrogen and maintained frozen (<-70C). Fetal carcasses were discarded without further evaluation. H.6.b. Fo Generation Rats Assigned to Natural Delivery On DL 5 the rats were sacrificed by carbon dioxide asphyxiation and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Blood, milk, heart and liver samples were collected from maternal rats on DL 5a. Gross lesions were retained in neutral buffered 10% formalin for possible future evaluation. Representative photographs of gross lesions are available in the raw data. On DL 5, each dam was removed from the nesting box and individually housed for approximately four hours. The dam was injected intravenously with one unit of oxytocin approximately five minutes before milk samples (at least 100 pL per sample) were collected. The samples were immediately frozen on dry ice and maintained frozen (<-20C). Following milk sample collection, blood samples (at least 4 mL each) were collected from the rats via the inferior vena cava and separated into two approximately equal aliquots. The first aliquot was placed into EDTA-coated tubes and the second aliquot was transferred into serum separator tubes. The samples were spun in a centrifuge and the resulting serum (divided into two aliquots) and plasma (one aliquot) were transferred into labeled polypropylene tubes. All samples were immediately frozen on dry ice and maintained frozen (<-20C). Following collection of milk and blood samples, the heart and liver of each dam were collected. The liver weight was recorded. A portion of each liver sample (right lateral lobe) was flash frozen in liquid nitrogen and maintained frozen (<-70C). The median a. See APPENDDC D, item 12. 000128 418-018:PAGE H-17 liver lobe was stored frozen (<-20C). The heart was excised and two cuts were made to allow proper fixation. The first cut started to the right of the ventral midline surface at the apex and extended anteriorly and ventrally to the pulmonary artery. The second cut was made starting to the left of the ventral midline surface at the apex and extended through the left ventricle to the ascending aorta. The cut heart and a section (approximately 0.3 to 0.5 cm wide) from the remaining portion of the liver were fixed in gluteraldehyde. The remaining portion of the liver was stored frozen (<-20C). Rats that did not deliver a litter were sacrificed on DG 25 and examined for gross lesions. Blood and tissue samples were not collected. Uteri were stained with 10% ammonium sulfide to confirm the absence of implantation sites(9) and retained in neutral buffered 10% formalin for possible future evaluation. Dams with no surviving pups were sacrificed after the last pup was found dead, missing or presumed cannibalized. Blood and tissue samples (serum, plasma, heart and liver) were collected as previously described. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed3. Rats that died or were sacrificed because of moribund condition were examined for the cause of death or moribund condition on the day the observation was made. The rats were examined for gross lesions. Representative photographs of gross lesions are available in the raw data. When possible (not precluded by autolysis), serum, plasma, heart and liver samples were retained as previously described for rats assigned to natural delivery. Pregnancy status and uterine contents of female rats were recorded. H.6.c. FI Generation Pups Pups that died before examination of the litter for pup viability were evaluated for vital status at birth. The lungs were removed and immersed in water. Pups with lungs that sank were identified as stillborn; pups with lungs that floated were identified as livebom, and to have died shortly after birth. Pups with gross lesions were preserved in Bouin's solution for possible future evaluation. When postmortem autolysis precluded these evaluations, it was noted on the litter observation data sheets. Pups found dead or sacrificed due to moribund condition on DLs 2 to 5 were examined for gross lesions and for the cause of the moribund condition or death. When postmortem autolysis precluded these evaluations it was noted on the litter observation data sheets. On DL 5, pups were sacrificed and examined for gross lesions. Gross lesions were preserved in neutral buffered 10% formalin. Necropsy included a single cross-section of the head at the level of the frontal-parietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Caps and labeled tubes were weighed (combined weight, to the nearest .001 gram) before (empty) and after retention of samples for subsequent use in pharmacokinetic analyses. a. See APPENDIX D, item 15. 0004)29 418-018-.PAGE H-18 Sample tubes were labeled with the study number, rat identification, date of collection, study day, sample identification and collection timepoint. Blood samples were collected via cardiac puncture from each pup. For Replicate 1, blood samples were pooled per litter, samples from male and female pups combined. Approximately 1 mL of blood was transferred into serum separator tubes; any remaining blood was transferred into EDTA-coated tubes. The samples were spun in a centrifuge and the resulting serum was divided into two aliquots of approximately 200 mL and approximately 300 mL of serum, and stored frozen (<-70C). Any resulting plasma was transferred into one aliquot and stored frozen (<-70C). For Replicate 2, blood samples were pooled (two litters per sample within each dosage group). Approximately 0.3 mL of blood was transferred into EDTA-coated tubes. The remaining blood was transferred into serum separator tubes. The samples were spun in a centrifuge and the resulting serum was divided into two aliquots and stored frozen (<-70C). Resulting plasma was transferred into one aliquot and stored frozen (<-70C). The liver from each pup was collected, pooled (by sex per litter) and the pooled weight recorded3. One liver from each litter pool was removed and fixed in gluteraldehyde for possible electron microscopy. The remaining livers in each litter pool were divided into three samples of equal number (when possible). Two of the samples were stored frozen (<-20C). The remaining sample was flash frozen in liquid nitrogen and maintained frozen (<-70C). The hearts were collected from the first two male and two female pups from each litter (pooled by sex per litter) and were fixed in gluteraldehyde. Thyroids were excised from each pup, pooled (by sex per litter). Thyroids were retained in neutral buffered 10% formalin. The remaining carcasses were discarded without further evaluation a. See APPENDDC D, item 16. 0001130 418-018:PAGE II-19 H.6.d. Sample Shipment and Analyses Sample Storage/Shipping Recipient Analysis Conditions Dams Assigned to Caesarean-Sectioning Serum - aliquot 1 <-20C Sponsor PFOS Serum - aliquot 2 <-20C AniLytics Total cholesterol, glucose, total & free T3, T4, TSH (Groups 1, 11, 12, 13), LDL, HDL, triglycerides3 Plasma <-20C Primedica Worcester Mevalonic acid (Groups 1,2, 3, 4, 11, 12 and 13) Liver - right lateral lobe <-70C Sponsor Possible biochemical Liver - median lobe <-20C Sponsor PFOS Liver - section Gluteraldehyde Sponsor Possible EM Liver - remainder <-20C AniLytics LDL, HDL, total cholesterol, triglycerides Fetuses Pooled serum - aliquot 1 <-20C Sponsor PFOS Pooled serum - aliquot 2 <-20C AniLytics Total cholesterol, glucose, total & free T3, T4,TSH (Groups 1, 11, 12, 13), LDL, HDL, triglycerides3 Pooled Plasma <-20C Primedica Worcester Mevalonic acid (Groups 1,2, 3, 4, 11, 12 and 13) Liver - 1 liver Gluteraldehyde Sponsor Possible EM Liver - 1/3 pooled livers <-70C Sponsor Possible biochemical Liver - 1/3 pooled livers <-20C Sponsor PFOS Liver - 1/3 pooled livers <-20C AniLytics LDL, HDL, total cholesterol, triglycerides Dams Assigned to Natural Delivery Milk <-20C AniLytics Total cholesterol Serum - aliquot 1 <-20C Sponsor PFOS Serum - aliquot 2 <-20C AniLytics Total cholesterol, glucose, total & free T3, T4, TSH (Groups 1,11, 12, 13), LDL, HDL, triglycerides3 Plasma <-20C Primedica Worcester Mevalonic acid (Groups 1 ,2 ,3 , 4, 11, 12 and 13) Liver - right lateral lobe <-70C Sponsor Possible biochemical Liver - median lobe <-20C Sponsor PFOS Liver - section Gluteraldehyde Sponsor Possible EM Liver - remainder <-20C AniLytics LDL, HDL, total cholesterol, triglycerides Heart Gluteraldehyde Sponsor Possible EM and light microscopy a. The first priority was analysis for total cholesterol and glucose. The second priority was for total and free T3, T4 and TSH levels. The third priority was analysis for LDL, HDL and triglycerides. nani 418-018:PAGE 11-20 Sample Storage/Shipping Recipient Analysis Conditions Pups - Replicate 1 Serum - aliquot 1 <-70C Sponsor PFOSa Serum - aliquot 2 <-70C AniLytics Total cholesterol, glucose, total & free T3, T4, TSH (Groups 1,11, 12, 13), LDL, HDL, triglyceridesb Plasma <-70C Primedica Worcester Mevalonic acid (Groups 1,2, 3,4, 11, 12 and 13) Pups - Replicate 2 Serum - aliquot 1 <-70C Sponsor PFOSa Serum - aliquot 2 <-70C AniLytics Total cholesterol, glucose, total & free T3, T4, TSH (Groups 1,11, 12, 13), LDL, HDL, triglyceridesb Plasma <-70C Primedica Worcester Mevalonic acid (Groups 1,2, 3,4, 11, 12 and 13) All Pups Liver - 1 liver per sex Gluteraldehyde Sponsor Possible EM Liver - 1/3 pooled livers <-70C Sponsor Possible biochemical Liver - 1/3 pooled livers <-20C Sponsor PFOS Liver - 1/3 pooled livers <-20C AniLytics LDL, HDL, total cholesterol, triglycerides Hearts - 2 males and 2 Gluteraldehyde Sponsor Possible EM and light females per litter0 microscopy Pooled Thyroids11 10% neutral buffered Testing Facility Possible histopathology formalin a. Analyses of serum were prioritized as follows: clinical chemistries first and then PFOS samples. b. The first priority was analysis for total cholesterol and glucose. The second priority was for total and free T3, T4 and TSH levels. The third priority was analysis for LDL, HDL and triglycerides. c. Histopathology was performed by Research Pathology Services, Inc. on the heart of one male and one female FI generation pup from each of four litters from the Vehicle Control group (Group 1) and 2 mg/kg PFOS group (Group 13). d. Histopathology was performed by Research Pathology Services, Inc. on the thyroid of one male and one female FI generation pup from each of four litters from the Vehicle Control group (Group 1) and 2 mg/kg PFOS group (Group 13). Results of these analyses are available in APPENDICES H and I. H.7. Data Collection and Statistical Analyses Data generated during the course of this study were recorded either by hand or using the Primedica Argus Automated Data Collection and Management System and the Vivarium Temperature and Relative Humidity Monitoring System. All data were tabulated, summarized and/or statistically analyzed using the Primedica Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microsoft Excel [part of Microsoft Office 97 (version SR-2)] and/or The SAS System (version 6.12). 004* 3* 418-018:PAGE 11-21 The following schematic represents the statistical analyses of the data: Type of Test3 I. Parametric A. Bartlett's Testc II. Nonparametricb A. Kruskal-Wallis Test (<75% ties) Significant at /><0.001 Not Significant Significant at p^O.05 Not Significant Nonparametric Analysis of Variance Dunn's Test Significant at ><0.05 Not Significant B. Fisher's Exact Test (>75% ties) Dunnett's Test III. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilities are reported as either p<0.05 or p<0.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. oobias 418-018'.PAGE 11-22 Group 1 (vehicle control) data were first statistically compared to Group 2 (mevalonic acid control) and Group 5 (cholesterol control) data. Group 1 (vehicle control) data were then compared to Groups 8, 9, 10, 11, 12 and 13 (0.4, 0.8, 1.0, 1.2, 1.6 and 2.0 mg/kg PFOS alone). Group 2 (mevalonic acid control) data were compared to Groups 3 and 4 (1.6 and 2.0 mg/kg PFOS + mevalonic acid) and Group 5 (cholesterol control) data were compared to Groups 6 and 7 (1.6 and 2.0 mg/kg PFOS + cholesterol, respectively). Clinical observations and other proportion data were analyzed using the Variance Test for Homogeneity of the Binomial Distribution(10). Continuous data (e.g., maternal body weights, body weight changes, feed consumption values, litter averages for percent male pups, pup body weights, pup mortality data) were analyzed using Bartlett's Test of Homogeneity of Variances0 0 and the Analysis of Variance02*, when appropriate [i.e., Bartlett's Test was not significant (/0.001)]. If the Analysis of Variance was significant (p<0.05), Dunnett's Test03*was used to identify the statistical significance of the individual groups. If the Analysis of Variance was not appropriate [i.e., Bartlett's Test was significant (p<0.001)], the Kruskal-Wallis Test04*was used, when less than or equal to 75% ties were present. In cases where the Kruskal-Wallis Test was statistically significant (p<0.05), Dunn's Method of Multiple Comparisons05*was used to identify the statistical significance of the individual groups. If there were greater than 75% ties, Fisher's Exact Test06*was used to analyze the data. Count data obtained at Caesarean-sectioning of the dams were evaluated using the procedures described above for the Kruskal-Wallis Test04*. 4>*M 34 418-018:PAGEIII-1 III. RESULTS A. Mortality (Summary - Table 1; Individual Data - Table 23) No deaths were attributable to the test or control articles. One rat in each of Groups 2 (mevalonic acid control), 3 (1.6 mg/kg PFOS + mevalonic acid) and 5 (cholesterol control) died as the result of intubation accidents or natural causes. Observations in these rats are described below. All other rats survived to scheduled sacrifice. Dam 10926 in Group 2 (mevalonic acid control) was moribund sacrificed on gestation day 12 (DG 12). Adverse clinical observations included excess salivation and soft or liquid feces on DG 11, and decreased motor activity, chromorhinorrhea, chromodacryorrhea and a red perioral substance on DG 12. This rat lost weight after DG 11. Feed consumption values were unremarkable. Necropsy revealed a perforation in the esophagus and tan discoloration of the left axillary area; all other tissues appeared normal. The litter consisted of 14 live embryos that appeared normal for their developmental ages. The death was attributed to an intubation accident. Rat 10936 in Group 3 (1.6 mg/kg PFOS + mevalonic acid) was found dead on DS 2. There were no adverse clinical observations before death. Necropsy revealed a dark red clotted material in the thoracic cavity; all other tissues appeared normal. The death was attributed to an intubation accident. Rat 11562 in Group 5 (cholesterol control) was moribund sacrificed on DS 16. Adverse clinical observations included decreased motor activity, pale appearance and coldness to touch on DS 16. Body weight was reduced on DS 15 compared to other rats in this dosage group. Feed consumption values were unremarkable. Necropsy revealed the small intestines were twisted and constricted at the jejunum and the jejunum contained a dark red fluid; all other tissues appeared normal. The death was attributed to natural causes. B. Clinical Observations (Summary - Table 1; Individual Data - Table 23) The numbers of rats with excess salivation, a perioral substance and chromorhinorrhea during gestation were significantly increased (p<0.01) in Group 5 (mevalonic acid control) as compared to Group 1 (vehicle control). These increases were attributed to an aversion to the taste of mevalonic acid; the incidence of these observations was comparable among the three mevalonic acid groups (Groups 2, 3 and 4). All other clinical observations during the precohabitation, gestation and lactation periods were considered unrelated to the test or control articles because; 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in one to four rats in a group. These observations included localized alopecia, dental problems (missing, broken or misaligned incisors), chromodacryorrhea, scabs on paws and limbs, bent tail, missing or swollen digit, swollen snout, red perivaginal substance, lacrimation, scab on mouth, tip of tail missing, ptosis, soft or liquid feces, rales, red substance in cage, labored breathing, scant feces, swollen ear, dehydration, emaciation, decreased motor activity, pale in appearance, cold to touch, and ulceration or scab on chest or limb. The significant increase 00M 35 418-018:PAGE III-2 (p<0.01) in the incidence of localized alopecia on the back during the gestation period in Group 11 (1.2 mg/kg PFOS alone) as compared to Group 1, was considered unrelated to the test article because it was not dosage dependent. C. Necropsy Observations (Summary - Table 2; Individual Data - Table 24) All necropsy observations were considered unrelated to the test or control articles because: 1) the incidences were not dosage-dependent; and 2) the observation occurred in only one rat in a group. These observations included misshapen kidneys, slight dilation of the pelvis of both kidneys, a calculus in the bladder and thickened bladder walls in one Group 2 (mevalonic acid control) rat and pup tissue in the stomach of one Group 7 (2 mg/kg PFOS + cholesterol) rat that was sacrificed because it had no surviving pups on day 2 of lactation (DL 2). Necropsy observations in rats that were found dead were described previously. D. Terminal Body Weights and Organ Weights and Ratios of Organ Weights to Terminal Body Weights (Summary - Table 3; Individual Data - Table 25) D.l. Caesarean-Sectioned Rats Terminal body weights of rats Caesarean-sectioned on DG 21 were significantly reduced (><0.05 or ><0.01) in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) as compared to Group 5 values (cholesterol control). The weight of the liver was significantly increased (><0.05) in Group 7. As a result of the reduced terminal weights in Groups 6 and 7 and the increased liver weight in Group 7, the ratios of the liver weight to the terminal body weight were significantly increased (><0.05 or /?<0.01) in these two groups, as compared to the cholesterol control. All other terminal body weights, liver weights and relative liver weights of rats Caesareansectioned on DG 21 were unaffected by PFOS or mevalonic acid. The ratio of liver weight to terminal body weight was significantly increased (><0.05) in Group 3 (1.6 mg/kg PFOS + mevalonic acid) as compared to the Group 2 value (mevalonic acid control), but was considered unrelated to the test or control articles because the increase was not dosage dependent. D.2. Naturally Delivered Rats Terminal body weights of naturally delivered rats were significantly reduced (><0.05 or ><0.01) in Group 13 (2 mg/kg PFOS alone) as compared to Group 1 (vehicle control), and in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) as compared to Group 5 values (cholesterol control). The ratios of the liver weights to the terminal body weights were significantly increased (p<0.05 or ><0.01) in Groups 9, 11 and 13 (0.8,1.2 and 2 mg/kg PFOS alone, respectively), compared to Group 1 (vehicle control), reflecting slight reductions in terminal body weights and/or slight increases in liver weights in these groups as compared to the Group 1 values. The liver weight was significantly increased (p<0.01) in Group 3 (1.6 mg/kg PFOS O0M3S 418-018:PAGE HI-3 + mevalonic acid) as compared to Group 2 (mevalonic acid control). The ratios of the liver weight to the terminal body weight were significantly increased (p<0.05 or /><0.01), in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) as compared to Group 2 (mevalonic acid control), and in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively), as compared to Group 5 (cholesterol control). E. Body Weights and Body Weight Changes (Figures 1 through 3; Summaries Tables 4 through 9; Individual Data - Tables 26 through 28) E.l. Precohabitation Body weight gains for the entire precohabitation period (DSs 1 to 42) were significantly reduced (p<0.05 or /><0.01) in Group 13 (2 mg/kg PFOS alone) as compared to Group 1 (vehicle control), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively), as compared to Group 5 (cholesterol control). During this period, gains were significantly reduced {p<0.05 or /?<0.01) in Groups 6 and 7 on DSs 22 to 29 and 29 to 36; and in Group 13 on DSs 29 to 36, as compared with the corresponding control group value. Additionally, body weight gains for Group 12 (1.6 mg/kg PFOS alone) were significantly reduced (p<0.05) on DSs 29 to 36 and body weight loss occurred in this group on DSs 36 to 42. As a result of these changes, body weights were significantly reduced (p<0.01) in Groups 6 and 7 on DSs 29, 36 and 42, as compared with Group 5. Body weight gains for the entire precohabitation period (DSs 1 to 42) in Group 5 (cholesterol control) were significantly increased (p<0.05) as compared to Group 1 (vehicle control). Premating body weights and body weight gains were unaffected by mevalonic acid and dosages of PFOS as high as 1.2 mg/kg. Significant increases or reductions (p<0.05 or /?<0.01) in body weight gains in Group 8 (0.4 mg/kg PFOS alone) on DSs 1 to 8 and 1 to 42, Group 12 (1.6 mg/kg PFOS alone) on DSs 1 to 8, Group 3 (1.6 mg/kg PFOS + mevalonic acid) on DSs 22 to 29 and 29 to 36, and Group 4 (2 mg/kg PFOS + mevalonic acid) on DSs 1 to 8 and 29 to 36 were not considered an effect of the test or control articles because they were not dosage-dependent and did not persist. E.2. Gestation Body weight gains for the entire gestation period (DGs 0 to 21) were significantly reduced (p<0.05) only in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol). Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) and Groups 9, 11, 12 and 13 (0.8, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively) had significantly reduced (p<0.05 or /?<0.01) body weight gains, compared to their corresponding control groups, for the first week of the gestation period (DGs 0 to 7). There were no significant differences among any of these dosage groups during the second week of gestation (DGs 7 to 14). Group 13 had a O0M37 418-018:PAGE III-4 significant increase (p<0.05) in body weight gain during the third week of gestation (DGs 14 to 21). Body weights were significantly reduced (p<0.05 or p<0.01) in Group 12 (1.6 mg/kg PFOS alone) on DGs 2 through 16, and in Group 13 (2 mg/kg PFOS alone) on DGs 0 through 19, compared to Group 1 (vehicle control). When PFOS was administered along with mevalonic acid, the effect on body weight was less severe and appeared over a shorter period than PFOS administered alone. Body weights were significantly reduced (p<0.05 or p<0.01) in Group 3 (1.6 mg/kg PFOS + mevalonic acid) on DGs 4 through 13, and in Group 4 (2 mg/kg PFOS + mevalonic acid) on DGs 7 through 10, compared to Group 2 (mevalonic acid control). The effect on body weight was more severe and appeared over a longer period when PFOS was administered along with cholesterol. Body weights were significantly reduced (p<0.01) in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DGs 0 through 21, compared to Group 5 (cholesterol control). Gestation body weights were significantly increased (p<0.05) in the cholesterol control group (Group 5) on DGs 5, 8 through 17 and 21, as compared to the vehicle control group (Group 1). Despite this effect of cholesterol alone, body weights in the groups administered PFOS with cholesterol (Groups 6 and 7) had significant reductions in gestation body weights. E.3. Lactation Body weight gains during the first five days of the lactation period (DLs 1 to 5) were significantly reduced (p<0.05 or/?<0.01) in Groups 9, 10, 12 and 13 (0.8, 1, 1.6 and 2 mg/kg PFOS alone, respectively). The average body weight on DL 5 was significantly reduced (p<0.05) in Group 13, compared to the vehicle control group value. When PFOS was administered along with mevalonic acid, the effect on body weight was less severe than PFOS administered alone. Body weight gains were significantly reduced (p<0.05) on DLs 1 to 5 in Group 3, but average body weights on DLs 1 and 5 were comparable among Groups 2, 3 and 4 (mevalonic acid control, 1.6 and 2 mg/kg PFOS + mevalonic acid, respectively). When PFOS was administered along with cholesterol, the effect on body weight was more severe. Body weights on DLs 1 and 5 were significantly reduced (p<0.Q5 or p<0.01) in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively), compared to Group 5 (cholesterol control), and a significant (p<0.05) body weight loss occurred on DLs 1 to 5 in Group 7. 000038 418-018:PAGE III-5 F. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values (Summaries - Tables 10 through 15; Individual Data - Tables 29 through 31) F.l. Precohabitation Absolute and relative feed consumption values were significantly reduced (p<0.05 or /?<0.01) in Group 13 (2 mg/kg PFOS alone) for the entire premating period (DSs 1 to 42) and within this period on DSs 8 to 15 (relative only), 22 to 29, 29 to 36 and 36 to 42, compared to the vehicle control group. Absolute and relative feed consumption values were also significantly reduced (/?<0.05) in Group 12 (1.6 mg/kg PFOS alone) during the last week of the premating period (DSs 36 to 42). Absolute and relative feed consumption values were unaffected by dosages of PFOS alone as high as 1.2 mg/kg. Absolute feed consumption was significantly increased (p<0.05) in Group 8 (0.4 mg/kg PFOS alone) on DSs 29 to 36; and absolute and relative feed consumption values were significantly increased (p<0.05) in Group 10 (1 mg/kg PFOS alone) on DSs 15 to 22. These increases in feed consumption values were not considered related to treatment because they were not dosage dependent and did not persist. When PFOS was administered along with mevalonic acid, the effect on feed consumption values was comparable to that of PFOS alone. Absolute feed consumption values were significantly reduced (p<0.05 or p<0.01) in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) on DSs 22 to 29 and 36 to 42, compared to Group 2 (mevalonic acid control). Relative feed consumption values were significantly reduced (p<0.05 or /><0.01) in Groups 3 and 4 on DSs 8 to 15, 15 to 22, 22 to 29 and 36 to 42 (Group 4 only). This effect of PFOS and mevalonic acid was reflected in the significantly reduced (><0.05 or p<0.01) absolute and relative feed consumption values for the entire premating period (DSs 1 to 42) in both groups. When PFOS was administered along with cholesterol, the effect on feed consumption values was more severe. Absolute and relative feed consumption values were significantly reduced (p<0.01) in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DSs 22 to 29, 29 to 36 and 36 to 42, as compared to Group 5 (cholesterol control). This effect of PFOS and cholesterol was reflected in the significantly reduced (p<0.05 or p<0.01) absolute and relative feed consumption values for the entire premating period (DSs 1 to 42) in both groups. Relative feed consumption values on DSs 8 to 15, 15 to 22, 22 to 29 and 1 to 42 were significantly increased (/?<0.05 or p<0.01) in the mevalonic acid control group (Group 2), as compared to the vehicle control group (Group 1). Absolute and relative feed consumption values were significantly increased (p<0.01) on DSs 29 to 36 and absolute feed consumption was significantly increased (p<0.05) on DSs 36 to 42 in the cholesterol control group (Group 5) as compared to the vehicle control group (Group 1). 009039 418-018-.PAGE m-6 F.2. Gestation Absolute and relative feed consumption values were significantly reduced (p<0.05 or p<0.01) during the first week of gestation (DGs 0 to 7) in Groups 9 through 12 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively). Absolute feed consumption continued to be significantly decreased (/?<0.05) in Group 13 on DGs 7 to 14, and relative feed consumption was significantly increased (p<0.05) in Group 12 on DGs 14 to 21, compared to the vehicle control group values. Absolute feed consumption values for the entire gestation period (DGs 0 to 21) were significantly reduced (p<0.05 or p<0.01) in Groups 12 and 13 (1.6 and 2 mg/kg PFOS alone, respectively), compared to the vehicle control group. When PFOS was administered along with mevalonic acid, effects on feed consumption values were comparable to the effects of administration of PFOS alone. Absolute feed consumption values were significantly reduced (p<0.01) in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) on DGs 0 to 7, 7 to 14 and 0 to 21, compared to Group 2 (mevalonic acid control). Relative feed consumption values were significantly reduced (p<0.05 or p<0.01) in Groups 3 and 4 on DGs 0 to 7 and 14 to 21. When PFOS was administered along with cholesterol, effects on feed consumption values were also comparable to the effects of administration of PFOS alone. Absolute feed consumption values were significantly reduced (p<0.01) in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DGs 0 to 7, 7 to 14, and 0 to 21, compared to Group 5 (cholesterol control). Relative feed consumption values were significantly reduced (/?<0.01) in Groups 6 and 7 on DGs 0 to 7 and significantly increased (p<0.05 or /?<0.01) in Groups 6 and 7 on DGs 14 to 21. F.3. Lactation Absolute feed consumption values during the first five days of the lactation period (DLs 1 to 5) were significantly reduced (/?<0.01) in Groups 12 and 13 (1.6 and 2 mg/kg PFOS alone, respectively). Relative feed consumption values for the same period were reduced in Groups 9 through 13, reaching statistical significance (p<0.05 or p<0.01) in Groups 9, 10, 11 and 12 (0.8, 1, 1.2 and 1.6 mg/kg PFOS alone, respectively), compared to Group 1 (vehicle control) values. When PFOS was administered along with mevalonic acid, effects on feed consumption values were also comparable to the effects of administration of PFOS alone. Absolute and relative feed consumption values were significantly reduced (p<0.01) in Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) on DLs 1 to 5, compared to the Group 2 (mevalonic acid control) value. When PFOS was administered along with cholesterol, effects on feed consumption values were comparable to the effects of administration of PFOS alone. Absolute and relative feed consumption values were significantly reduced (p<0.01) in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DLs 1 to 5, as compared to Group 5 (cholesterol control). 418-018:PAGE m-7 G. Mating and Fertility (Summary - Table 16; Individual Data - Table 32) The number of days in cohabitation, the fertility and pregnancy indices (number of pregnancies per number of rats that mated and rats in cohabitation, respectively), and the number of rats with confirmed mating dates during the first and second week of cohabitation were comparable in the thirteen dosage groups. The number of days in cohabitation was significantly reduced (p<0.01) for Group 6 (1.6 mg/kg PFOS + cholesterol), as compared to Group 5 (cholesterol control). This finding was not considered treatment-related because it was not dosage-dependent. H. Caesarean-Sectioning and Litter Observations (Summaries - Tables 17 and 18; Individual Data - Tables 33 through 35) Caesarean-sectioning observations on DG 21 were based on eight pregnant dams in each of Groups 1 through 7, 12 and 13. No Caesarean-sectioning or litter parameters were affected by dosages up to 2 mg/kg PFOS alone or administered with either cholesterol or mevalonic acid. There were no biologically important differences in the litter averages for corpora lutea, implantations, live and dead fetuses, early and late resorptions, percent dead or resorbed conceptuses, percent live male fetuses or pooled fetal body weights. No dam had a litter in which there were no live fetuses and all placentae appeared normal. The average number of resorptions (total and early), the number of dams with any resorptions and the percentage of dead or resorbed conceptuses per litter were significantly reduced (p<0.05 or p<0.01) for Group 13 (2 mg/kg PFOS alone), compared to Group 1 (vehicle control) values. These findings were not considered treatment-related because an increase, rather than a decrease, in the incidence of resorption is the expected effect of a toxicant. The percentage of dead or resorbed conceptuses per litter was significantly increased (p<0.05) in Group 7 (2 mg/kg PFOS + cholesterol); although this value is above the range observed historically at this Testing Facility2, the value is comparable to the concurrent vehicle control group (Group 1) and reflects the low value in the cholesterol control group. I. Natural Delivery and Litter Observations (Summaries - Tables 19 and 20; Individual Data - Tables 36 through 38) Totals of 17 to 20 rats were pregnant and delivered litters in each of the 13 dosage groups. The duration of gestation was significantly reduced {p<0.05 or p<0.01) in Groups 9 through 13 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively), as compared to the corresponding control a. See APPENDIX K (HISTORICAL CONTROL DATA). 000041 418-018:PAGE III-8 values. The number of dams with all pups dying on DLs 1 to 5 was increased in Groups 13, 3, 4, 6 and 7; the increase was significant at ><0.01 in Groups 13,4 and 7. The pup viability index (number of live pups on DL 5 per number of livebom pups on DL 1) was significantly reduced (><0.05 or ><0.01) in Groups 12 and 13 (1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7(1.6 and 2 mg/kg PFOS + cholesterol, respectively). The numbers of pups that died or were presumed cannibalized on DL 1 and DLs 2 to 5 was significantly increased (p<0.05 or ><0.01) in Groups 12, 13, 4, 6 and 7. The numbers of surviving pups per litter were significantly reduced (><0.05 or ><0.01) on DLs 2, 3, 4 and 5 in Groups 12, 13, 3, 4, 6 and 7. A dosage-dependent pattern of reduced pup body weight was evident on DLs 1, 2, 3, 4, 5 and/or 1 to 5 in each group administered the test article. Groups 11,12 and 13 (1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively) and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) had significantly reduced (><0.01) pup body weights on most weighing days (DLs 1 through 5). When average pup weight was normalized for litter size, Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) and Groups 8, 9, 10, 11, 12 and 13 (0.4, 0.8, 1, I. 2, 1.6 and 2 mg/kg/day PFOS alone, respectively) were also significantly reduced (><0.05 or p<0.01) from Group 2 (mevalonic acid control), Group 5 (cholesterol control) and Group 1 (Vehicle Control), respectively, over the same days of lactation. All other natural delivery and litter parameters were unaffected by dosages of PFOS as high as 2 mg/kg administered alone or in combination with mevalonic acid or cholesterol. The average number of implantation sites, the gestation index and the numbers of livebom and stillborn pups were comparable across all 13 dosage groups. The number of dams with stillborn pups was significantly increased (><0.01) in Group 8 (0.8 mg/kg PFOS alone) and significantly reduced (><0.05) in Groups 10, 11, 12 and 13 (1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively). These significant differences were not considered treatment-related because they were not dosage-dependent. The significant reduction (><0.01) in the percentage of male pups on DL 3 in Group 4 was related to the increased pup mortality in this dosage group and not a direct effect of PFOS with mevalonic acid. J. Clinical Observations from Birth to Day 5 Postpartum and Necropsy Observations (Summaries - Tables 21 and 22; Individual Data Tables 39 and 40) Adverse clinical and necropsy observations associated with reduced pup viability and potential reduction in maternal care occurred in Groups 11, 12 and 13 (1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol). Increased numbers of litters with pups that were cold to touch occurred in Groups 11, 12, 13,4, 6 and 7. The number of litters with pups not nursing was increased in Groups 12, 13, 3, 4, 6 and 7; the increase was significant at ><0.01 in Group 7. OOOOii 418-018:PAGE III-9 Necropsy observations in pups that were stillborn or found dead included increases in the numbers of pups with no milk in the stomach in Groups 9, 10, 11, 12 and 13 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mg/kg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol). The incidence of this observation was significantly increased (p<0.01) in Group 6, as compared with Group 5 (cholesterol control). All other clinical and necropsy observations in the FI generation pups were considered unrelated to the test article because: 1) the incidences were not dosage-dependent; or 2) the observation occurred in only one litter. Clinical observations included decreased motor activity, gasping, labored breathing, not nesting, exophthalmos, swollen limb, tail problems (missing, portion missing and tip black) and no milk in stomach. The necropsy observation of protruding tongue was significantly reduced (p<0.01) in Groups 8, 9, 10, 11, 12 and 13, reflecting the single occurrence of this observation in the vehicle control group (Group 1). A single pup in the mevalonic acid control group (Group 2) had no oral opening, no eyes and displaced ears; as a result, the incidence of this observation was significantly reduced (p<0.01) in Groups 3 and 4. One pup in Group 8 (0.4 mg/kg PFOS alone) had a tan area on the median lobe of the liver at necropsy on DL 5. All other pups appeared normal at necropsy on DL 5. K. Blood and Tissue Analyses (See APPENDICES H, I, J and K) No significant difference between control groups (Groups 1, 2 and 5) for any of the parameters. K .l. Gestation Day 21 K.l.a. Dams Clinical chemistry parameters were generally unaffected in the DG 21 dams that were treated with 1.6 or 2 mg/kg PFOS alone. Clinical Chemistry Values (mg/d L) of PFOS alone Parameter Cholesterol Glucose LDL-Dir HDL-Dir Group 1 84.3 13.3 98.9 11.7 7.1 5.0 43.3 19.0 (Vehicle) (8 ) (8 ) (8 ) (8 ) Group 12 78.1 10.5 104.9 7.8 10.8 4.4 43.5 11.2 1 .6 mg/kg (8 ) (8 ) (8 ) (8 ) Group 13 72.3 6.3 96.3 18.0 11.3 3.6 41.0 11.7 2 l"gftg (8 ) (8 ) (8 ) (8 ) Triglycerides 313.5 233.6 (8 ) 194.3 65.2 (8 ) 201.0 87.5 (8 ) Mevalonic Acid 47.7 63.8 (8 ) 20.6 5.7 (8 ) 86.2 177.0 (8 ) The total and free thyroid hormones levels for T3 and T4 in dams administered 1.6 or 2 mg/kg PFOS alone were significantly reduced (p<0.01), compared to the vehicle control group. 00004 418-018:PAGE III-10 Thyroid Hormone Values of PFOS a one Total T4 Total T3 Free T3 Parameter ug/dL ng/dL pg/mL Group 1 0.584 0.482 73.979 21.234 0.579 0.209 (Vehicle) (8 ) (8 ) (8 ) Group 12 0 .0 00 0 .0 0 0 ** 47.503 10.268** 0.140 0.117** 1 .6 mg/kg (8) (8 ) (7) Group 13 0 .0 0 0 0 .0 0 0 ** 42.440 8.271** 0.157 0.161** 2 mg/kg (8 ) (8 ) (7) TSH ng/mL 1,815 0.794 (8 ) 2.346 1.263 (7) 1.514 0.973 (7) Free T4 ng/dL 0.299 0.092 (8 ) 0.066 0.038** (8 ) 0.055 0.023** (8 ) Liver cholesterol levels were significantly decreased (><0.05 or ><0.01) in DG 21 dams administered 1.6 or 2 mg/kg PFOS, respectively, compared to the vehicle control group. Other liver clinical chemistry parameters were generally unaffected in the dams that were treated with 1.6 or 2 mg/kg PFOS alone. Liver Clinical Chemistry Values (mg/gm) of PFOS alone Parameter Cholesterol LDL-Dir HDL-Dir Group 1 3.0 0.38 0.5 0.27 0.5 0.12 (Vehicle) (8 ) (8 ) (8 ) Group 12 2 .6 0 .2 2 * 0.7 0.24 0.6 0 .10 1 .6 mg/kg (8 ) (8 ) (8 ) Group 13 2.4 + 0.26** 0 .6 0 .12 0.5 0.06 2 mg/kg (8 ) (8 ) (8 ) Triglycerides 8.0 1.66 (8 ) 7.7 0.71 (8 ) 6 .6 0.82 (8 ) The clinical chemistry parameter of low density lipoprotein (LDL) was significantly increased (><0.05 or p<0.01), albeit not dosage-dependent, in the DG 21 dams that were coadministered mevalonic acid and PFOS. Clinical Chemistry Values (mg/dL) of PFOS plus Mevalonic Acid Parameter Cholesterol Glucose LDL-Dir HDL-Dir Triglycerides Mevalonic Acid Group 2 92.5 14.6 108.5 11.9 2.9 3.6 32.4 20.0 453.9 307.8 1663.8 415.6 (Mevalonic) (8 ) (8 ) (8 ) (8 ) (8 ) (8 ) Group 3 91.0 21.4 108.1 15.4 11.8 9.0** 35.1 12.1 305.9 85.1 1369.8 240.1 1 .6 mg/kg (8 ) (8 ) (8 ) (8 ) (8 ) (8 ) Group 4 88.9 23.1 106.0 14.0 9.6 5.0* 47.8 16.2 243.6 270.8 1482.5 521.9 ___2 m8/kg (8 ) (8 ) (8 ) (8) (8 ) (8 ) The total and free thyroid hormones levels for T3 and T4 in DG 21 dams coadministered mevalonic acid and 1.6 or 2 mg/kg PFOS were significantly reduced (><0.05 or ><0.01), compared to the vehicle control group. Significantly different from control group; ><0.05. Significantly different from control group; ><0.01. 09P 044 418-018:PAGE III-11 Thyroid Hormone Values of PFOS plus Mevalonic Acid Total T4 Total Tj Free T3 TSH Parameter ug/dL ng/dL pg/mL ng/mL Group 2 0.414 + 0.510 73.571 14.314 0.514 0.203 2.627 1.177 (Mevalonic) (8 ) (8) (7) (6 ) Group 3 0.003 0.007* 54.043 6.670** 0.313 0.146* 1.889 0.391 1 .6 mg/kg (8 ) (8 ) (8 ) (7) Group 4 0.000 0.000** 46.408 8.895** 0.184 0.170** 2.375 1.086 2 mg/kg (8 ) (8 ) (8 ) (8 ) Free T4 ng/dL 0.296 0.098 (8 ) 0.069 0.044** (8 ) 0.073 0.035** (8 ) Liver cholesterol and triglyceride levels were significantly reduced (p<0.01) in dams coadministered mevalonic acid and 1.6 or 2 mg/kg PFOS. Liver LDL and HDL levels were significantly increased in dams coadministered mevalonic acid and 1.6 or 2 mg/kg PFOS. Liver Clinical Chemistry Values (mg/gm) of PFOS plus Mevalonic Acid Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 2 3.0 0.34 0.5 0.24 0.5 0.08 9.8 1.45 (Mevalonic) (8 ) (8 ) (8 ) (8 ) Group 3 2.5 0.12** 0 .8 0.08** 0.6 0.03** 7.5 0.55** 1 .6 mg/kg (8 ) (8 ) (8 ) (8 ) Group 4 2.4 0.30** 0 .8 0 .1 2 ** 0.6 0.05 7.0 1.35** 2 mg/kg (8 ) (8 ) (8 ) (8 ) Clinical chemistry parameters were generally unaffected in the DG 21 dams that were treated with cholesterol and PFOS. Clinical C emistry Values (mg/d Parameter Cholesterol Glucose Group 5 89.6 19.6 100.4 20.2 (Cholesterol) (8 ) (8 ) Group 6 79.3 9.4 98.5 18.5 1 .6 mg/kg (8 ) (8 ) Group 7 77.9 11.5 106.4 17.9 ___2 ?g/kg___ (8 ) (8 ) of PFOS plus Cholesterol LDL-Dir HDL-Dir 8.5 5.2 28.0 11.5 (8 ) (8 ) 9.9 3.9 41.0 13.6 (8 ) (8 ) 12 .0 8.6 46.6 18.4 (8 ) (8 ) Triglycerides 348.8 218.6 (8 ) 191.9 79.3 (8 ) 200.9 190.1 (8 ) The total and free thyroid hormones levels for T3 and T4 in DG 21 dams coadministered cholesterol and 1.6 or 2 mg/kg PFOS were significantly reduced (p<0.01), compared to the vehicle control group. Significantly different from control group; p<0.05. Significantly different from control group; p<0.01. 004045 418-018:PAGE III-12 Thyroid Hormone Values of PFOS plus Cholesterol Total T, Total T3 Free T3 Parameter ug/dL ng/dL pg/mL Group 5 0.540 0.529 76.900 21.899 0.694 0.256 (Cholesterol) (8) (8) (7) Group 6 0.013+0.035** 52.031 9.331** 0.235 0.123** 1.6 mg/kg (8) (8) (8) Group 7 2 mg/kg 0.000 0.000** 49.020 8.527** 0.246 0.180** (8) (8) (8) TSH ng/mL 2.544 0.759 (5) 1.452 0.691 (6) 2.013 0.854 (6) Free T4 ng/dL 0.281 0.125 (7) 0.134 0.086** (8) 0.110 0.082** (8) Liver triglyceride levels were significantly reduced (p<0.01) in DG 21 dams coadministered cholesterol and 1.6 or 2 mg/kg PFOS. Liver LDL and HDL levels were increased or significantly increased in dams coadministered cholesterol and 1.6 or 2 mg/kg PFOS. Liver cholesterol parameters were generally unaffected in the dams that were treated with 1.6 or 2 mg/kg PFOS plus Cholesterol. Liver Clinical Chemistry Va ues (mg/gm) of PFOS plus Cholesterol Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 5 3.0 0.30 0.3 0.28 0.5 0.13 9.6 1.00 (Cholesterol) (8) (8) (8) (8) Group 6 2.7 0.29 0.8 0.23** 0.6 + 0.10 7.7 0.72** 1.6 mg/kg (8) (8) (8) (8) Group 7 2.7+0.21 0.8 0.18** 0.6 0.09 7.5 0.70** 2 mg/kg (8) (8) (8) (8) PFOS concentrations were significantly increased (p<0.05) in the sera of DG 21 dams administered 1.6 or 2 mg/kg PFOS and in dams coadministered Cholesterol or Mevalonic acid and 1.6 or 2 mg/kg PFOS. PFOS concentrations were significantly increased (p<0.05) in the livers of dams administered 1.6 or 2 mg/kg PFOS. Significantly different from control group; p < 0.01. 0000146 418-018:PAGE III-13 PFOS Levels in DG 21 Dam Sera and Liver Sera" Liver3 Parameter (ug/mL) (ppm) Group 1 (Vehicle) 0.0 0.00 (6) N.A. Group 2 (Mevalonic) 0.0 0.02 (6) N.A. Group 3 1.6 mg/kg 55.8 8.88* (6) N.A. Group 4 2 mg/kg 82.4 22.55* (6) N.A. Group 5 (Cholesterol) 0.0 0.00* (6) N.A. Group 6 1.6 mg/kg 79.9 15.48* (6) N.A. Group 7 2 mg/kg 370.8 214.36* (6) N.A. Group 8 0.4 mg/kg N.A. N.A. Group 9 0.8 mg/kg N.A. N.A. Group 10 1 mg/kg N.A. N.A. Group 11 1.2 mg/kg N.A. N.A. Group 12 1.6 mg/kg 142.2 24.24* (6) N.A. Group 13 2 mg/kg 124.7 19.71* (6) N.A. N.A. = Not Analyzed a = Sponsor selected the subsets to be analyzed. K.l.b. DG 21 Fetuses The clinical chemistry parameters of cholesterol and low density lipoprotein (LDL) were significantly increased (p<0.05 or p<0.01) in the DG 21 fetuses from dams that were treated with 1.6 or 2 mg/kg PFOS alone. Significantly different from control group; p < 0.05. 000047 418-018:PAGE HI-14 Clinical Chemistry Values (mg/dL) of PFOS alone Parameter Cholesterol Glucose LDL-Dir HDL-Dir Group 1 (Vehicle) Group 12 1.6 mg/kg Group 13 2 mg/kg 50.8 8.0 (8) 61.3 8.5* (8) 61.5 8.0* (8) 63.8 5.9 (8) 76.1 11.8 (8) 73.9 15.5 (8) 27.3 8.0 (8) 44.9 7.5** (8) 45.4 9.8** (8) 13.0 1.9 (8) 14.5 1.5 (8) 14.0 1.2 (8) Triglycerides 34.4 5.3 (8) 27.9 8.2 (8) 27.0 9.8 (8) Mevalonic Acid 54.5 40.5 (7) 67.0 53.0 (8) 104.8 73.9 (8) The level of free T4 was significantly reduced (p<0.01) in the DG 21 fetuses from dams that were treated with 1.6 or 2 mg/kg PFOS alone. Thyroid Hormone Values of PFOS alone Total T4 Total Tj Free T3 Parameter ug/dL ng/dL pg/mL Group 1 0.000 0.000 0.423 0.845 0.000 0.000 (Vehicle) (7) (4) (0) Group 12 0.000 0.000 0.000 0.000 0.000 0.000 1.6 mg/kg (8) (8) (1) Group 13 0.000 0.000 0.925 1.147 0.000 0.000 ___2 mg'kg__ (8) (6) (0) TSH ng/mL 0.000 0.000 (0) 0.000 0.000 (0) 1.160 0.000 (1) Free T4 ng/dL 0.050 0.000 (1) 0.000 0.000** (3) 0.000 0.000** (5) Liver HDL clinical chemistry parameters were significantly increased (p<0.01) in DG 21 fetuses from dams administered 1.6 or 2 mg/kg PFOS alone. Other liver clinical chemistry levels were generally unaffected in the fetuses of dams that were treated with 1.6 or 2 mg/kg PFOS alone. Liver Clinical Chemistry Values (mg/gm) of PFOS alone Parameter Cholesterol LDL-Dir HDL-Dir Group 1 3.4 0.38 0.000 0.0000 0.01 0.009 (Vehicle) (8) (8) (8) Group 12 3.4 0.38 0.000 0.0000 0.02 0.006* 1.6 mg/kg (8) (8) (8) Group 13 3.2 0.34 0.000 0.0000 0.02 0.015* _____2 m ?/kg____ (8) (8) (8) Triglycerides 4.9 0.63 (8) 5.9 0.93 (8) 5.6 0.91 (8) The clinical chemistry parameters of cholesterol and low density lipoprotein (LDL) were significantly increased (p<0.01) and triglycerides were significantly decreased (p<0.01) in the DG 21 fetuses from dams that were treated with mevalonic acid and 1.6 and/or 2 mg/kg PFOS. Significantly different from control group; p<0.05. Significantly different from control group; p<0.01. 00004ft 418-018:PAGE III-15 Clinical C lemistry Va lues (mg/dl of PFOS plus Mevalonic Acid Parameter Cholesterol Glucose LDL-Dir HDL-Dir Triglycerides Group 2 (Mevalonic) 53.1 5.5 (8) 63.4 15.0 29.4 5.3 (8) (8) 13.9 1.2 (8) 36.4 8.7 (8) Group 3 68.3 13.4** 69.6 6.2 48.3 8.1** 16.1 3.0 1.6 mg/kg (8) (8) (8) (8) 31.8 7.1 (8) Group 4 2 mg/kg 63.1 11.7 74.5 12.3 46.9 9.4** 15.5 1.9 25.1 8.1** (8) (8) (8) (8) (8) Mevalonic Acid 18971.4 5681.5 (7) 16562.5 7160.4 (8) 20175.0 2830.8 (8) The level of free T4 was significantly reduced (/?<0.01) in the DG 21 fetuses from dams that were treated with mevalonic acid and 1.6 or 2 mg/kg PFOS. Thyroid Hormone Values of PFOS plus Mevalonic Acid Total T4 Total T3 Free T3 TSH Parameter ug/dL ng/dL pg/mL ng/mL Group 2 0.000 0.000 2.810 4.867 (Mevalonic) (7) (3) (0) (0) Group 3 0.000 0.000 1.096 1.379 0.000 0.000 1.6 mg/kg Group 4 (8) 0.000 0.000 (7) (1) 2.928 2.747 0.000 0.000 (0) 2 mg/kg (8) (6) (2) (0) Free T4 ng/dL 0.070 0.000 (1) 0.000 0.000** (2) 0.000 0.000** (4) Liver cholesterol, HDL and triglyceride levels were significantly increased (p<0.05 or p<0.01) in the DG 21 fetuses of dams that were treated with 1.6 or 2 mg/kg PFOS plus Mevalonic Acid. Liver Clinical Chemistry Va ues (mg/gm) of PFOS plus Mevalonic Acid Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 2 3.6 0.49 0.000 0.0000 0.01 0.009 5.4 1.25 (Mevalonic) (8) (8) (8) (8) Group 3 4.5 0.48** 0.000 0.0000 0.04 0.019** 8.0 1.16** 1.6 mg/kg (8) (8) (8) (8) Group 4 5.1 0.82** 0.000 0.0000 0.03 0.018* 9.6 2.39** _____2 m g/kg . (8) (8) (8) (8) The clinical chemistry parameters of cholesterol, low density lipoprotein (LDL) and high density lipoprotein (HDL) were significantly increased (p<0.05 or p<0.01) in the DG 21 fetuses from dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS. Significantly different from control group; p<0.01. D 00B 4B 418-018:PAGE III-16 Clinical C emistry Values (mg/dL of PFOS p us Cholesterol Parameter Cholesterol Glucose LDL-Dir HDL-Dir Group 5 45.1 4.3 70.4 7.9 24.1 5.1 12.4 0.9 (Cholesterol) (8 ) (8 ) (8 ) (8 ) Group 6 59.1 11.8** 77.4 19.8 43.9 5.4** 14.4 1.6* 1 .6 mg/kg (8 ) (8) (8 ) (8 ) Group 7 64.0 10.3** 70.3 10.3 45.6 8.0** 14.4 1.7* 2 mg/kg (8 ) (8 ) (8 ) (8 ) Triglycerides 31.8 7.6 (8 ) 25.4 13.6 (8 ) 30.0 5.7 (8 ) The level of free T4 was significantly reduced (p<0.01) in the DG 21 fetuses from dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS. Thyroid Hormone Values of PFOS p us Cholesterol Parameter Total T4 ug/dL Total T3 ng/dL Free T3 pg/mL TSH ng/mL Group 5 0 .0 0 0 0 .0 0 0 1.143 1.306 0 .0 0 0 0 .0 0 0 0 .0 0 0 0 .0 0 0 (Cholesterol) (8 ) (6 ) (2 ) (0 ) Group 6 0 .0 0 0 0 .0 0 0 0.760 1.389 0 .0 0 0 0 .0 0 0 0.040 0.000 1 .6 mg/kg Group 7 2 mg/kg (7) 0 .0 0 0 0 .0 0 0 (6 ) (5) 0.653 1.132 (3) (2 ) 0 .0 0 0 0 .0 0 0 (1) (1) 0 .0 0 0 0 .0 0 0 (0 ) Free T4 ng/dL 0.037 0.015 (3) 0 .0 0 0 0 .0 0 0 ** (4) 0 .0 0 0 0 .0 0 0 ** (2 ) Liver cholesterol and triglyceride levels were increased or significantly increased (p<0.01) in the DG 21 fetuses from dams coadministered cholesterol and 1.6 or 2 mg/kg PFOS. Liver LDL and HDL levels were generally unaffected in the fetuses of dams that were treated with 1.6 or 2 mg/kg PFOS plus Cholesterol. Liver Clinical Chemistry Values (mg/gm) of PFOS plus Cholesterol Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 5 3.6 0.29 0 .0 0 0 0 .0 0 0 0 0.02 0.019 5.5 0.73 (Cholesterol) (8 ) (8 ) (8) (8 ) Group 6 1 .6 mg/kg 4.4 0.40** (8 ) 0 .0 0 0 0 .0 0 0 0 (8 ) 0.03 0.010 (8 ) 7.8 1.23** (8 ) Group 7 4.0 0.68 0 .0 0 0 0 .0 0 0 0 0.04 0.023 7.1 1.15** _____2 mg/k____ (8 ) (8 ) (8 ) (8) PFOS concentrations were significantly increased (/?<0.01) in the pooled sera of DG 21 fetuses administered 1.6 or 2 mg/kg PFOS and in fetuses coadministered Cholesterol or Mevalonic acid and 1.6 or 2 mg/kg PFOS. PFOS concentrations were significantly increased (p<0.05) in the livers of fetuses administered 1.6 or 2 mg/kg PFOS. Significantly different from control group; p< 0.05. Significantly different from control group; p < 0 .0 1. 000050 418-018:PAGE III-17 PFOS Levels in Pooled DG 21 Fetal Sera and Livers Seraa Livera Parameter (ug/mL) (ppm) Group 1 (Vehicle) 0 .0 0 .0 0 (6 ) N.A. Group 2 (Mevalonic) 0 .1 0.08 (6 ) N.A. Group 3 1 .6 mg/kg 207.2 25.76** (6 ) N.A. Group 4 2 mg/kg 251.0 55.86** (6 ) N.A. Group 5 (Cholesterol) 0.1 0.03 (6 ) N.A. Group 6 1 . 6 mg/kg 154.5 32.43** (6 ) N.A. Group 7 2 mg/kg 165.7 50.13** (6 ) N.A. Group 8 0.4 mg/kg N.A. N.A. Group 9 0 .8 mg/kg N.A. N.A. Group 10 1 mg/kg N.A. N.A. Group 11 1 .2 mg/kg N.A. N.A. Group 12 1 . 6 mg/kg 141.8 33.10** (6 ) N.A. Group 13 169.8 37.91** ____ 2 N.A. = Not Analyzed (6 ) N.A. a = Sponsor selected subsets to be analyzed. K.2. Lactation Day 5 K.2.a. Dams The clinical chemistry parameter of serum cholesterol was significantly decreased (p<0.01) in the lactating dams that were treated with 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS alone, compared to the vehicle control group. The clinical chemistry parameter of glucose was significantly increased (p<0.01) in lactating dams treated with 2 mg/kg PFOS alone and the clinical chemistry parameter of triglycerides was significantly decreased (p<0.05 or p<0.01) in lactating dams treated with 1.6 or 2 mg/kg PFOS alone, compared to the vehicle control group. Significantly different from control group; p<0.05. Significantly different from control group; /?<0.01. 000051 418-018:PAGE III-18 Clinical Chemistry Values (mg/dL) of PFOS alone Parameter Group 1 (Vehicle) Group 8 0.4 mg/kg Group 9 0 .8 mg/kg Group 10 1 mg/kg Group 11 1 .2 mg/kg Cholesterol [Blood] 77.6 + 15.2 (19) 65.0 13.2** (2 0 ) 59.1 13.3** (17) 58.5+11.8** (19) 59.9 + 11.7** (18) Cholesterol [Milk] 96.7 53.9 (19) 71.1 62.8 (2 0 ) 68.2 52.6 (17) 59.8 51.2 (18) 46.8 41.5 (18) Group 12 1 .6 mg/kg Group 13 2 mg/kg 56.6 14.0** (17) 59.6 10.2** (19) 64.6 30.4 (13) 74.0 41.7 (5) Glucose 166.1 28.4 (19) 169.6 26.7 (2 0 ) 163.0 17.3 (17) 171.9 19.9 (19) 172.5 17.7 (18) 178.7 19.0 (17) 189.2 32.2** (19) LDL-Dir 4.0 3.0 (19) 3.2 2.3 (2 0 ) 3.2 2.1 (17) 3.7 1.9 (19) 2.8 2.2 (18) 3.6 2.6 (17) 4.6 1.8 (19) HDL-Dir 58.2 20.1 (19) 51.2 17.2 (2 0 ) 47.5 14.6 (17) 49.0 11.0 (19) 48.9 16.2 (18) 49.5 10.3 (17) 53.1 9.4 (19) Triglyceride s 152.4 97.6 (19) 130.1 78.3 (2 0 ) 127.7 66.5 (17) 101.2 52.7 (19) 141.1 74.3 (18). 92.2 58.9* (17) 85.5 35.0** (19) Mevalonic Acid 31.0 19.8 (18) Not Evaluated Not Evaluated Not Evaluated 83.1 178.8** (18) 32.2 20.9 (16) 29.6 14.4 (15) The total and free T4 levels in lactating dams administered 0.4 (free T4only), 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced (/?<0.05 or p<0.01) and total and free T3 levels in lactating dams administered 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced (p<0.05 or p<0.01), compared to the vehicle control group. Thyroid Hormone Values of PFOS alone Total T4 Total Tj Free T3 Parameter ug/dL ng/dL pg/mL Group 1 1.454 0.660 74.674 18.977 0.638 0.297 (Vehicle) (19) (19) (19) Group 8 0.808 0.414 72.912 13.470 0.698 0.246 0.4 mg/kg GO) GO) GO) Group 9 0.596 0.443* 0.8 mg/kg (8) 63.780 6.684 (8) 0.588 0.152 (8) Group 10 0.728 0.243** 1mg/kg (9) 62.348 13.180 (9) 0.552 0.230 (9) Group 11 0.283 0.320** 52.869 14.974** 0.411 0.265* 1.2 mg/kg (18) (18) (18) Group 12 0.272 0.172** 47.032 19.971** 0.256 0.230** 1.6 mg/kg (17) (17) (17) Group 13 0.235 0.147** 53.275 17.270** 0.353 0.236** 2 m /kg (19) (19) (18) TSH ng/mL 1.633 0.992 (18) 1.142 0.226 (9) 1.437 0.918 (7) 1.110 0.523 (8) 1.447 1.029 (17) 1.665 0.768 (17) 1.533 0.681 (18) Free T4 ng/dL 0.788 0.219 (19) 0.462 0.114** (10) 0.279 0.060** (8) 0.277 0.095** (9) 0.260 0.122** (18) 0.253 0.067** (17) 0.259 0.078** (18) Liver triglyceride levels were significantly increased {p<0.01) in lactating dams that were treated with 1.6 or 2 mg/kg PFOS. All other liver clinical chemistry parameters were generally unaffected in the DL 5 dams that were treated with 1.6 or 2 mg/kg PFOS alone. Significantly different from control group; p < 0.05. Significantly different from control group; p<0.01. 000052 418-018:PAGE HI-19 Liver Clinical Chemistry Values (mg/gm) of PFOS alone Parameter Cholesterol LDL-Dir HDL-Dir Group 1 2.4 1.29 0.00 0.014 0.13 0.094 (Vehicle) (19) (19) (19) Group 8 0.4 mg/kg 2.6 0.67 (2 0 ) 0.01 0.034 (2 0 ) 0.22 0.115 (2 0 ) Group 9 2.4 0.59 0.05 0.093 0.23 0.130 0 .8 mg/kg (17) (17) (17) Group 10 2.5 0.54 0.00 0.004 0.20 0.093 1 mg/kg (19) (19) (19) Group 11 2.5 0.83 0.05 0.106 0.26 0.135 1 .2 mg/kg (18) (18) (18) Group 12 2.6 0.79 0.01 0.038 0.21 0.104 1 .6 mg/kg (17) (17) (17) Group 13 2.9 1.08 0.03 0.054 0.22 0.117 ____ 2 mg/kg____ (19) (19) (19) Triglycerides 7.7 2.90 (19) 8.4 1.70 (2 0 ) 8 .6 2.47 (17) 11.8 12.02 (19) 9.7 3.44 (18) 13.4 7.20** (17) 16.0 5.38** (19) The clinical chemistry parameters of serum cholesterol was significantly decreased (p<0.01) in the lactating dams that were treated with mevalonic acid and 1.6 or 2 mg/kg PFOS and cholesterol in the milk was significantly increased (p<0.05) in the lactating dams that were treated with mevalonic acid and 2 mg/kg PFOS. Clinical Chemistry Va ues (mg/dL) of PFOS plus Mevalonic Acid Cholesterol Cholesterol Parameter [Blood] [Milk] Glucose LDL-Dir HDL-Dir Group 2 80.5 15.6 81.4 52.4 174.3 39.2 3.7 2.9 63.7 18.1 (Mevalonic) (18) (17) (18) (18) (18) Group 3 1 .6 mg/kg 63.6 14.6** (18) 70.5 46.2 (1 2 ) 174.9 24.1 3.4 2.0 54.2 12.1 (18) (18) (18) Group 4 2 mg/kg 67.3 11.2** 188.0 114.6* 187.6 30.9 3.7 3.1 55.0 10.1 (19) (2 ) (19) (19) (19) Tri glycerides 135.6 58.6 (18) 103.3 43.5 (18) 118.7 50.7 (19) Mevalonic Acid 201.4 135.9 (18) 1123.0 2 0 0 0 .2 * (18) 18662.4 70304.8* (19) The levels of total and free T4 were significantly reduced (p<0.01) in the lactating dams that were treated with mevalonic acid and 1.6 or 2 mg/kg PFOS. Thyroid Hormone Values of PFOS plus Mevalonic Acid Total T4 Total T3 Free T3 Parameter ug/dL ng/dL pg/mL Group 2 2.608 1.083* 88.864 19.532 0.822 0.295 (Mevalonic) (5) (5) (5) Group 3 0.182 0.243** 65.752 6.320 0.695 0.131 1 .6 mg/kg (4) (4) (4) Group 4 0.332 0.245** 76.158 10.814 0.638 0.208 2 mg/kg (6 ) (6 ) (5) TSH ng/mL 1.962 1.655 (5) 1.228 1.076 (4) 1.366 0.751 (5) Free T4 ng/dL 0.894 0.248 (5) 0.270 0.067** (4) 0.222 0.076** (5) Liver triglyceride levels were significantly increased (p<0.01) in the lactating dams that were treated with 2 mg/kg PFOS plus Mevalonic Acid. All other liver clinical chemistry parameters were generally unaffected in the DL 5 dams that were treated with 1.6 or 2 Significantly different from control group; p<0.05. Significantly different from control group; p<0.01. 004053 418-018:PAGE ID-20 mg/kg PFOS plus Mevalonic Acid. Liver Clinical Chemistry Values (mg/gm) of PFOS plus Mevalonic Acid Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 2 2.3 1.00 0.01 0.041 0.16 0 .1 2 2 7.5 2.05 (Mevalonic) (18) (18) (18) (18) Group 3 2.2 0.65 0.02 0.064 0.19 0.134 10.3 6.24 1 .6 mg/kg (18) (18) (18) (18) Group 4 2.7 1.29 0.01 0.031 0.18 0.129 15.2 5.49** 2 mg/kg (19) (19) (19) (19) Levels of cholesterol in the blood and triglycerides were significantly decreased (/?<0.01) in lactating dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS and the blood level of glucose was significantly increased (p<0.05) in lactating dams that were treated with cholesterol and 2 mg/kg PFOS. Clinical Chemistry Values (mg/dL) of PFOS p us Cholesterol Cholesterol Cholesterol Parameter fBloodl [Milk] Glucose LDL-Dir HDL-Dir Group 5 75.9 17.5 80.2 57.9 168.8 29.2 3.8 3.0 55.6 19.1 (Cholesterol) (17) (17) (17) (17) (17) Group 6 56.2 12.2** 81.9 52.5 170.2 2 0 .8 4.4 2.8 48.6 11.1 1 .6 mg/kg (2 0 ) (12 ) (2 0 ) (2 0 ) (2 0 ) Group 7 58.0 15.2** 57.8 3.8 190.6 28.5* 4.9 2.4 50.9 13.3 2 mg/kg (19) (4) (19) (19) (19) Triglycerides 146.1 64.8 (17) 87.2 32.1** (2 0 ) 85.0 36.1** (19) The level of total and free T3 and T4 were significantly reduced (p<0.05 or p<0.01) in the lactating dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS. Thyroid Hormone Values of PFOS plus Cholesterol Total T4 Total T3 Free T3 Parameter ug/dL ng/dL pg/mL Group 5 2.087 0.758 91.763 14.299 0.887 0.151 (Cholesterol) (6 ) (6 ) (6 ) Group 6 0.272 0.304** 65.235 11.143** 0.510 0.150* 1 .6 mg/kg (6 ) (6 ) (5) Group 7 0.134 0.195** 65.714 18.542* 0.602 0.306* __ 2mAg (5) (5) (5) TSH ng/mL 0.810 0.422 (6 ) 1.280 0.665 (5) 1.884 1.147 (5) Free T4 ng/dL 0.927 0.294 (6 ) 0.272 0.039** (6 ) 0.242 0.074* (5) Liver triglyceride levels were significantly increased (p<0.05) in lactating dams coadministered cholesterol and 2 mg/kg PFOS. All other liver clinical chemistry parameters were generally unaffected in the DL 5 dams that were treated with 1.6 or 2 mg/kg PFOS plus Mevalonic Acid. Significantly different from control group; p<0.05. Significantly different from control group; p<0.01. 000054 418-018:PAGE III-21 Liver Clinical Chemistry Values (mg/gm) of PFOS plus Cholesterol Parameter Cholesterol LDL-Dir HDL-Dir Group 5 2.1+0.70 0.00 0.005 0.16 0.096 (Cholesterol) (17) (17) (17) O ko1+> p Group 6 1.6 mg/kg Group 7 2 mg/kg 2.3 0.69 (20) 2.3 0.61 (19) 0.01 0.036 (20) 0.00 0.000 (19) (20) 0.19 0.078 (19) Triglycerides 7.6 1.68 (17) 10.1 4.68 (20) 11.0 4.83* (19) PFOS Levels in DL 5aDam Sera and Livers Sera0 Liverb Parameter (ug/mL) (ppm) Group 1 0 .0 0 .0 1 0.5 0.87 (Vehicle) (6 ) (6 ) Group 2 (Mevalonic) 12.7 27.37 (6 ) N.A. Group 3 1 . 6 mg/kg 127.0 24.41* (6 ) N.A. Group 4 2 mg/kg 189.2 48.20* (6 ) N.A. Group 5 (Cholesterol) 0.7 1.62 (6 ) N.A. Group 6 1 . 6 mg/kg 104.3 14.57* (6 ) N.A. Group 7 2 mg/kg 144.8 7.60* (6 ) N.A. Group 8 27.2 18.72* 47.9 5.15* 0.4 mg/kg (6 ) (4) Group 9 0 .8 mg/kg 42.6 6.93* (6 ) N.A. Group 10 1 mg/kg 52.3 26.02* (6 ) N.A. Group 11 1 .2 mg/kg 8 6 .0 1 0 .2 2 * (6 ) N.A. Group 12 169.0 32.12* 110.2 13.43* 1 . 6 mg/kg (6 ) (5) Group 13 134.0 26.60* 145.8 20.18* 2 mg/kg (6 ) (6 ) N.A. = Not Analyzed a = Some dams were sacrificed due to no surviving pups on various days, b = Sponsor selected subsets to be analyzed. Significantly different from control group; p<0.05. 000055 418-018:PAGE I-22 K.2.b. LD 5 FI Generation Pups The clinical chemistry parameters measured were not significantly different in the FI generation pups from lactating dams that were treated with 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS alone, compared to the vehicle control group. Clinical Chemistry Values (mg/dL) of PFOS alone Parameter Cholesterol Glucose LDL-Dir Group 1 112.5 10.6 255.8 61.8 33.5 8.7 (Vehicle) Group 8 0.4 mg/kg (12 ) 111.9 11.3 (15) (12 ) 244.5 60.1 (15) (12 ) 29.1 7.4 (15) Group 9 114.8 11.1 220.3 56.2 31.5 7.6 0 .8 mg/kg (13) (13) (13) Group 10 113.1 16.8 210.0 47.0 29.4 5.3 1 mg/kg (14) (14) (14) Group 11 124.7 + 23.4 217.2 60.6 34.8 15.0 1 .2 mg/kg (14) (14) (14) Group 12 132.3 33.9 2 2 2 .0 2 0.8 40.3 20.1 1 .6 mg/kg (6 ) (6 ) (6 ) Group 13 128.0 38.3 229.3 17.2 44.7 31.6 2 mg/kg (3) (3) (3) HDL-Dir 29.8 7.9 (12 ) 32.7 6.9 (15) 38.1 11.0 (13) 36.3 6 .6 (14) 31.9 5.8 (14) 36.0 + 7.0 (6 ) 34.3 4.0 (3) Triglycerides 214.2 51.8 (1 2 ) 234.9 95.0 (15) 179.2 66.5 (13) 197.8 64.4 (14) 230.3 78.5 (14) 231.8 52.7 (6 ) 213.0 56.3 (3) Mevalonic Acid 22.6+5.3 (9) Not Evaluated Not Evaluated Not Evaluated 18.7 4.0 (8 ) 20.6 4.0 (4) (0 ) The free and total T4 levels in DL 5 pups from lactating dams administered 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced (/?<0.01), compared to the vehicle control group. The thyroid stimulating hormone (TSH) was significantly increased (p<0.05 or p<0.01) at 1 and 1.6 mg/kg PFOS, but not in a dosage-dependent fashion. Thyroid Hormone Values of PFOS alone Total T4 Total T3 Parameter ug/dL ng/dL Group 1 0.543 0.221 54.383 17.965 (Vehicle) (12) (12 ) Group 8 0 .0 0 0 0 .0 0 0 ** 55.777 18.934 0.4 mg/kg (10) (9) Group 9 0 .0 0 0 0 .0 0 0 ** 48.825 17.720 0 .8 mg/kg (8 ) (8) Group 10 0.017 0.050** 47.710 8.613 1 mg/kg (9) (7) Group 11 0.005 0.014** 44.715 21.746 1 .2 mg/kg Group 12 (13) 0.005 0.012** (1 1 ) 32.708 7.941 1 .6 mg/kg (6 ) (6 ) Group 13 0 .0 0 0 0 .0 0 0 ** 32.777 12.162 2 m/k___ (3) (3) Free T3 pg/rnL 0.039 0.086 (9) 0.016 0.036 (5) 0 .0 0 0 0 .0 0 0 (5) 0 .0 0 0 0 .0 0 0 (4) 0.003 0.009 GO) 0.004 0.009 (5) 0.050 0.087 (3) TSH ng/mL 1.018 0.166 (8 ) (0 ) (0 ) 2.360 0.000** (1 ) 1.010 0.246 (5) 1.450 0.339* (5) 1.500 0.000 (1 ) Free T4 ng/dL 0.090 0.024 (1 1 ) 0.018 0.009** (8 ) 0.014 0.018** (7) 0.016 0 .0 1 0 ** (7) 0.011 0.009** (1 1 ) 0 .0 10 0.006** (6 ) 0 .0 1 0 0 .0 1 0 ** (3) Liver triglyceride and LDL levels were decreased or significantly decreased (p<0.05 or /?<0.01) in DL 5 female pups from lactating dams administered 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS. All other liver clinical chemistry parameters were generally unaffected in the female pups of lactating dams that were treated with 0.4 to 2 mg/kg PFOS alone. Significantly different from control group; p<0.05. Significantly different from control group; p < 0.01. 000056 418-018:PAGE 01-23 Female Pups Liver Clinical Chemistry Values (mg/gm) of PFOS alone Parameter Cholesterol LDL-Dir HDL-Dir Group 1 3.8 0.66 0.012 0.0264 0.10 0.025 (Vehicle) (18) (18) (18) Group 8 3.9 0.48 0 .0 0 0 0 .0 0 2 2 ** 0.09 0.031 0.4 mg/kg (2 0 ) (2 0 ) (2 0 ) Group 9 3.8 0.30 0 .0 0 0 0 .0 0 0 0 ** 0.09 0.038 0 .8 mg/kg (17) (17) (17) Group 10 4.0 0.83 0 .0 0 0 0 .0 0 0 0 ** 0.08 0.035 1 mg/kg (18) (18) (18) Group 11 3.8 0.62 0 .0 0 0 0 .0 0 0 0 ** 0.09 0.037 1 .2 mg/kg (17) (17) (17) Group 12 1 .6 mg/kg 3.5 0.32 (10 ) 0.003 0.0095 GO) 0.11 0.032 (10 ) Group 13 2 mg/kg 3.8 0.62 (4) 0 .0 0 0 0 .0 0 0 0 (4) 0.09 0.017 (4) Triglycerides 18.1 7.94 (18) 14.0 6.18 (2 0 ) 14.0 4.67 (17) 12.8 4.56* (18) 12.0 6.85** (17) 11.4 3.11 * (10 ) 10.8 2.04 (4) Liver triglyceride levels were decreased or significantly decreased (p<0.05 or p<0.01) in DL 5 male pups from lactating dams administered 0.4, 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS. All other liver clinical chemistry parameters were generally unaffected in the male pups whose dams were treated with 0.4 to 2 mg/kg PFOS alone. Male Pups Liver Clinical Chemistry Values (mg/gm) of PFOS alone Parameter Cholesterol LDL-Dir HDL-Dir Group 1 3.5 0.49 0.003 0.0118 0.08 0.019 (Vehicle) (18) (18) (18) Group 8 3.9 0.83 0 .0 0 0 0 .0 0 0 0 0.08 0.043 0.4 mg/kg (2 0 ) (2 0 ) (2 0 ) Group 9 3.6 0.56 0 .0 0 0 0 .0 0 0 0 0.08 0.041 0 .8 mg/kg (17) (17) (17) Group 10 3.7 0.76 0.003 0.0097 0.10 0.030 1 mg/kg (18) (18) (18) Group 11 3.6 0.55 0 .0 0 0 0 .0 0 0 0 0.10 0.042 1 .2 mg/kg (18) (18) (18) Group 12 3.3 0.52 0.003 0.0067 0.09 0.030 1 .6 mg/kg GO) GO) (10 ) Group 13 3.4 0.26 0.005 0.0100 0.09 0.033 2 mg/kg (4) (4) (4) Triglycerides 17.3 8.05 (18) 15.4 + 6.76 (2 0 ) 14.4 6.46 (17) 11.1 3.21** (18) 10.9 4.80** (18) 11.4 7.55* GO) 7.4 1.54* (4) Levels of cholesterol, low density lipoproteins (LDL) and triglycerides in the blood were significantly increased (jp<0.05 or p<0.01) in DL 5 pups from lactating dams that were treated with mevalonic acid and 1.6 mg/kg PFOS. Significantly different from control group; ><0.05. Significantly different from control group; p<0.01. 000057 418-018:PAGE III-24 Clinical Chemistry Values (mg/dL) o PFOS plus Mevalonic Acid Parameter Cholesterol Glucose LDL-Dir HDL-Dir Triglycerides Group 2 115.4 7.6 241.4 88.4 35.3 7.5 35.4 9.0 193.2 69.9 (Mevalonic) (1 2 ) (12 ) (1 2 ) (12 ) (1 2 ) Group 3 157.0 23.7** 273.3 65.1 50.8 20.0* 33.0 4.9 302.3 62.7** 1 .6 mg/kg (7) (7) (7) (7) (7) Group 4 0 .00 0.000 0.0 0 0.000 0 .00 0.000 0.00 0.000 0.00 0.000 ___2 m/kg___ (0 ) (0 ) (0 ) (0 ) (0 ) Mevalonic Acid 335.3 229.2 (ID 270.8 85.8 (7) 195.2 203.4 (2 ) Levels of total T4 were significantly decreased (p<0.01) in DL 5 pups from lactating dams that were treated with mevalonic acid and 1.6 mg/kg PFOS. Thyroid Hormone Values of PFOS plus Mevalonic Acic Total T4 Total T-i Free T3 TSH Parameter ug/dL ng/dL pg/mL ng/mL Group 2 0.486 0.227 83.420 31.7202 0.497 0.490 0.430 0.212 (Mevalonic) (5) (5) (3) (2 ) Group 3 0 .0 0 0 0 .0 0 0 ** 70.633 24.823 0 .00 0.000 0.680 0 .0 00 1 .6 mg/kg (3) (3) (0 ) (1 ) Group 4 2 mg/kg NO VALUES FOR THIS GROUP Free T4 ng/dL 0.090 0.056 (5) 0.005 0.007 (2 ) Liver clinical chemistry parameters were generally unaffected in the DL 5 female pups of lactating dams that were treated with 1.6 PFOS plus Mevalonic Acid. There were no liver samples for female pups of lactating dams that were treated with 2 mg/kg PFOS plus Mevalonic Acid. Female Pups Liver Clinical Chemistry Values (mg/gm) of PFOS plus Mevalonic Acid Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 2 3.8 0.64 0.008 0.0218 0.11 0.039 18.4 8.93 (Mevalonic) (18) (18) (18) (18) Group 3 3.3 0.86 0.002 0.0067 0.08 0.030 14.1 6.00 1 .6 mg/kg (9) (9) (9) (9) Group 4 2 mg/kg NO VALUES FOR THIS GROUP Liver clinical chemistry parameters were generally unaffected in the DL 5 male pups of lactating dams that were treated with 1.6 PFOS plus Mevalonic Acid. There were no liver samples for male pups of lactating dams that were treated with 2 mg/kg PFOS plus Mevalonic Acid. * Significantly different from control group; p<0.05. ** Significantly different from control group; p<0.01. a. Group 2 significantly different from Group 1; p<0.01. 000058 418-018:PAGE HI-25 Male Pups Liver Clinical Chemistry Values (mg/gm) of PFOS plus Mevalonic Acid Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 2 3.6 + 0.69 0.003 0.0096 0.09 + 0.026 13.0 7.38 (Mevalonic) (18) (18) (18) (18) Group 3 3.3 + 0.68 0.008 0.0253 0.09 0.037 13.6 5.38 1 .6 mg/kg (10 ) GO) GO) (1 0 ) Group 4 ___ ? mg/kg___ NO VALUES FOR THIS GROUP Levels of low (LDL) and high (HDL) density lipoproteins were significantly increased (p<0.05 or p<0.01) in the pups from lactating dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS and the blood level of triglycerides was significantly decreased (p<0.05) in the pups from dams that were treated with cholesterol and 2 mg/kg PFOS. Clinical C emistry Values (mg/dL) of PFOS plus Cholesterol Parameter Cholesterol Glucose LDL-Dir HDL-Dir Group 5 117.8 9.1 263.8 56.4 26.8 1 2 .2 28.7 7.7 (Cholesterol) (1 2 ) (1 2 ) (1 2 ) (12 ) Group 6 130.0 + 18.5 247.8 48.9 41.0 11.4* 39.2 3.6** 1 .6 mg/kg (8 ) (8 ) (8 ) (8 ) Group 7 134.8 26.1 223.2 20.2 55.2 22.5** 47.2 12.8** 2 mg/kg (5) (5) (5) (5) Triglycerides 321.3 208.8 (1 2 ) 232.9 74.4 (8 ) 120.4 36.6* (5) Levels of total T3 and free T4 were significantly decreased (p<0.05 or p<0.01) in pups from lactating dams that were treated with cholesterol and 1.6 and/or 2 mg/kg PFOS. Thyroid Hormone Values of PFOS plus <Cholesterol Total T4 Total T3 Free T3 Parameter ug/dL ng/dL pg/mL Group 5 0.323 0.273a 78.405 17.178* 0.270 0.240 (Cholesterol) (6 ) (6 ) (2 ) Group 6 0 .0 0 0 0 .0 0 0 40.840 4.072** 0 .0 0 0 0 .0 0 0 1 .6 mg/kg (3) (3) (1 ) Group 7 0 .0 0 0 0 .0 0 0 25.900 0.000* 0 .0 0 0 0 .0 0 0 2 mg/kg (1 ) (1 ) (0 ) TSH ng/mL 1.380 0.000a (1) 0 .0 0 0 0 .0 0 0 (1 ) 0 .0 0 0 0 .0 0 0 (0 ) Free T4 ng/dL 0.064 0.027 (5) 0 .0 0 0 0 .0 0 0 * (2 ) 0 .0 0 0 0 .0 0 0 (0 ) Liver clinical chemistry parameters were generally unaffected in the DL 5 female pups of lactating dams that were treated with 1.6 or 2 mg/kg PFOS plus Mevalonic Acid. Female Pups Liver Clinical Chemistry Values (mg/gm) of PFOS plus Cholesterol Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 5 3.8 0.56 0.011 0.0271 0.09 0.040 17.9 7.29 (Cholesterol) (17) (17) (17) (17) Group 6 3.4 0.54 0 .0 0 0 0 .0 0 0 0 0.08 0.031 12.9 4.36 1 .6 mg/kg Group 7 (1 1 ) 4.0 0.14 (1 1 ) 0 .0 0 0 0 .0 0 0 0 (1 1 ) 0.10 0.037 (1 1 ) 13.5 5.16 ____ 2 mgflg____ (4) (4) (4) (4) * Significantly different from control group; p<0.05. ** Significantly different from control group; p<0.01. a. Group 5 significantly different from Group 1; p<0.01. 000059 418-018:PAGE m-26 Liver triglyceride levels were significantly reduced (/?<0.05 or p<0.01) in DL 5 male pups of dams coadministered cholesterol and 1.6 or 2 mg/kg PFOS. All other liver clinical chemistry parameters were generally unaffected in the male pups of lactating dams that were treated with 1.6 or 2 mg/kg PFOS plus Mevalonic Acid. Male Pups Liver Clinical Chemistry Values (mg/gm) of PFOS plus Cholesterol Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 5 3.7 + 0.45 0.004 0.0100 0.08 0.029 18.3 6.80 (Cholesterol) (17) (17) (17) (17) Group 6 1.6 mg/kg Group 7 2 mg/kg 3.4 0.45 (8) 4.1 0.73 (3) 0.000 0.0000 (8) 0.000 0.0000 (3) 0.10 0.031 (8) 0.08 0.035 (3) 10.5 3.95** (8) 10.6 4.12* (3) PFOS Levels in Pooled DL 5 Fetal Sera and Livers Seraa Liver3 Parameter (ug/mL) (ppm) Group 1 0 .0 0 .0 2 BQL (Vehicle) (6 ) (6 ) Group 2 (Mevalonic) 2.8 3.12 (6 ) N.A. Group 3 1 .6 mg/kg 123.7 18.58* (3) N.A. Group 4 2 mg/kg N.A. N.A. Group 5 (Cholesterol) 0.2 0.19 (6 ) N.A. Group 6 1 . 6 mg/kg 108.5 7.78 (2 ) N.A. Group 7 2 mg/kg Group 8 155.0 0.00 (1) 36.2 3.84 N.A. 73.4 30.83 0.4 mg/kg (6 ) (6 ) Group 9 0 .8 mg/kg 53.1 27.67 (6 ) N.A. Group 10 1 mg/kg 84.5 17.52 (6 ) N.A. Group 11 1 .2 mg/kg 146.7 162.50** (6 ) N.A. Group 12 1 . 6 mg/kg N.A. 274.0 127.80** (5) Group 13 138.0 0.00 245.2 93.36** 2 mg/kg N.A. = Not Analyzed (1) (4) BQL = Below Quantitation Limit a = Sponsor selected subsets to be analyzed. Significantly different from control group; p<0.05. Significantly different from control group; p<0.01. 000060 418-018:PAGE HI-27 L. Histopathology of Hearts and Thyroids of FI Generation Pups (See APPENDIX J) The type and incidence of the histomorphologic observations in the heart and thyroid of the male and female pups from dams of the vehicle control and 2 mg/kg/day dosage groups are shown below: Dose Group Sex Dam No. No. Pups THYROID No. examined No. Normal HEART No. examined No. Normal 1 M 10909 1 1 1 1 1 13 M 11059 1 1 1 1 1 1 F 10909 1 1 1 1 1 13 F 11059 1 1 1 1 1 No microscopic changes were observed; all sections examined were within normal histologic limits. 000061 418-018:PAGE III-28 REFERENCES 1. Study Design as Modification of: U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Federal Register, September 22, 1994, Vol. 59, No. 183. 2. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. 3. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standardfor Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 4. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. 5. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 6. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 7. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Crl:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research.) 8. Institute of Laboratory Animal Resources (1996). Guidefor the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 9. Salewski, E. (1964). Farbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. 10. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 240-241. 11. Sokal, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. 12. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. Q IM tt FINAL REPORT ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 FINAL REPORT DATE: 16 DECEMBER 2002 TITLE - ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS ARGUS STUDY NUMBER: 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 TABLE OF CONTENTS SUBJECT PAGE I. SUMMARY AND CONCLUSION 1-1 A. Methods 1-1 B. Results 1-4 C. Conclusion I-10 . DESCRIPTION OF TEST PROCEDURES n-i A. Conduct of Study n-i B. Test Article Information n-4 C. Control Articles Information n-4 D. Vehicle Information n-5 E. Test Article Preparation and Storage Conditions n-6 F. Test System n-7 G. Husbandry n-9 H. Methods n-n m. RESULTS m -i A. Mortality m -i B. Clinical Observations m -i C. Necropsy Observations 000#G4 m-2 SUBJECT PAGE D. Terminal Body Weights and Organ Weights and Ratios of Organ Weights to Terminal Body Weights III-2 E. Body Weights and Body Weight Changes IH-3 F. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values IU-5 G. Mating and Fertility ffi-7 H. Caesarean-Sectioning and Litter Observations EU-7 I. Natural Delivery and Litter Observations IU-7 J. Clinical Observations from Birth to Day 5 Postpartum and Necropsy Observations m-8 K. Blood and Tissue Analyses EQ-9 L. Histopathology of Hearts and Thyroids FIGeneration Pups DI-27 REFERENCES HI-28 APPENDIX A - REPORT FIGURES Figure 1. Body Weights - Fo Generation Female Rats- Groups 1, 8 through 13A-l Figure 2. Body Weights - Fo Generation Female Rats- Groups 2, 3 and 4A-2 Figure 3. Body Weights - Fo Generation Female Rats- Groups 5, 6 and 7A-3 APPENDIX B - REPORT TABLES Table 1. Clinical Observations - Summary - Fo Generation Female Rats B-l Table 2. Necropsy Observations - Summary - Fo Generation Female Rats B-9 Table 3. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight - Summary - Fo Generation Female Rats B -l2 Table 4. Body Weights - Premating - Summary - Fo Generation Female Rats B -l5 Table 5. Body Weight Changes - Premating - Summary - Fo Generation Female Rats B -l8 hi 000065 SUBJECT PAGE Table 6. Maternal Body Weights - Gestation - Summary - Fo Generation Female Rats B-21 Table 7. Maternal Body Weight Changes - Gestation - Summary Fo Generation Female Rats B-27 Table 8. Body Weights - Lactation - Summary - Fo Generation Female Rats B-30 Table 9. Body Weight Changes - Lactation - Summary - Fo Generation. Female Rats B-33 Table 10. Absolute Feed Consumption Values (g/day) - Premating - Summary - Fo Generation Female Rats B-36 Table 11. Relative Feed Consumption Values (g/kg/day) - Premating - Summary - Fo Generation Female Rats B-39 Table 12. Maternal Absolute Feed Consumption Values (g/day) - Gestation Summary - Fo Generation Female Rats B-42 Table 13. Maternal Relative Feed Consumption Values (g/kg/day) - Gestation - Summary - Fo Generation Female Rats B-45 Table 14. Absolute Feed Consumption Values (g/day) - Lactation - Summary Fo Generation Female Rats B-48 Table 15. Relative Feed Consumption Values (g/kg/day) - Lactation - Summary - Fo Generation Female Rats B-51 Table 16. Mating and Fertility - Summary - Fo Generation Female Rats B-54 Table 17. Caesarean-Sectioning Observations - Summary - Fo Generation Female Rats B-57 Table 18. Litter Observations (Caesarean-Delivered Fetuses) - Summary Fo Generation Female Rats B-60 Table 19. Natural Delivery Observations - Summary - Fo Generation Female Rats B-63 Table 20. Litter Observations (Naturally Delivered Pups) - Summary FI Generation Litters B-66 Table 21. Clinical Observations from Birth to Day 5 Postpartum - Summary FI Generation Pups IV 000066 B-75 SUBJECT PAGE Table 22. Necropsy Observations - Summary - FI Generation Pups B-78 Table 23. Clinical Observations - Individual Data - Fo Generation Female Rats B-81 Table 24. Necropsy Observations - Individual Data - Fo Generation Female Rats B-102 Table 25. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight - Individual Data - Fo Generation Female Rats B-122 Table 26. Body Weights - Premating - Individual Data - Fo Generation Female Rats B-135 Table 27. Maternal Body Weights - Presumed Gestation - Individual Data Fo Generation Female Rats B-148 Table 28. Maternal Body Weights - Lactation - Individual Data - Fo Generation Female Rats B-174 Table 29. Feed Consumption Values - Premating - Individual Data Fo Generation Female Rats B-187 Table 30. Maternal Feed Consumption Values - Presumed Gestation Individual Data - Fo Generation Female Rats B-200 Table 31. Maternal Feed Consumption Values - Lactation - Individual Data Fo Generation Female Rats B-213 Table 32. Mating and Fertility and Days in Cohabitation - Individual Data Fo Generation Female Rats B-226 Table 33. Caesarean-Sectioning Observations - Individual Data - Fo Generation Female Rats B-239 Table 34. Litter Observations (Caesarean-Delivered Fetuses) - Individual Data - Fo Generation Female Rats B-242 Table 35. Fetal Vital Status - Individual Data - Fo Generation Female Rats B-245 Table 36. Natural Delivery, Implantation Sites, and Pup Viability and Sex Individual Data - Fo Generation Female Rats B-248 Table 37. Pup Body Weights Pooled by Litter from Birth to Day 5 Postpartum - Individual Data - FI Generation Litters B-261 v 000067 SUBJECT PAGE Table 38. Pup Vital Status and Sex from Birth to Day 5 Postpartum - Individual Data - FI Generation Pups B-274 Table 39. Clinical Observations from Birth to Day 5 Postpartum - Individual Data - FI Generation Pups B-287 Table 40. Necropsy Observations - Individual Data - FI Generation Pups B-292 APPENDIX C - PROTOCOL AND AMENDMENTS C-l to C-67 APPENDIX D - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY D -l to D-6 APPENDIX E - CERTIFICATE OF ANALYSIS E-l to E-2 APPENDIX F - HOMOGENEITY AND CONCENTRATION ANALYSIS F-l toF-21 APPENDIX G - TEMPERATURE AND RELATIVE HUMIDITY REPORTS G-l toG-5 APPENDIX H - BIOANALYTICAL REPORTS (ANILYTICS) H-l to H-125 APPENDIX I - BIOANALYTICAL REPORTS (PRIMEDICA WORCESTER, SPONSOR AND SRI) I-1 to 1-444 APPENDIX J - HISTOPATHOLOGY REPORT J-l to J-7 APPENDIX K - HISTORICAL CONTROL DATA K-l to K-8 APPENDIX L - STATEMENT OF THE STUDY DIRECTOR L-l APPENDIX M - QUALITY ASSURANCE STATEMENT M-l toM-2 vi Q0Q 06S 418-018:PAGE III-29 13. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-1121. 14. Sokal, R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 15. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241-252. 16. Siegel, S. (1956). Nonparametric Statistics for the Behavioral Sciences, McGraw- Hill, New York, pp. 96-104. 000069 APPENDIX A REPORT FIGURES MN7 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) BODY WEIGHTS Fo GENERATION FEMALE RATS Figure 1 - Groups 1,8 -1 3 450 V GROUP 1 425 O 400 GROUP 8 375 GROUP 9 350 O GROUP 10 I 325 LU 300 275 250 225 i r u 1j vr I i.j T ..iw/m, I j * * * . Ji l? l * *m ' *i*** ** r m * ii ** ** ** X GROUP 11 >- GROUP 12 ri GROUP 13 *p < 0 .0 5 (from group 1) 200 \ 15 22 2!9 3!6 4 la ^ il I H H H H 10 11 12 1*3 14 15 16 17 18 19 20 21 Ik **p < 0 .0 1 (from group 1) DAY OF STUDY DAY OF GESTATION DAY OF LACTATION a. Last value recorded before cohabltaton. 418-018:PAGE A-l PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) BODY WEIGHTS Fo GENERATION FEMALE RATS Figure 2 - Groups 2,3,4 450 I I I I 425 ' GROUP 2 400 L7J GROUP 3 375 350 CD I- I 325 CD UJ 5 300 GROUP 4 *p<0.05 (from group 2) **p<0.01 (from group 2) 418-018:PAGE A-2 275 250 225 X nm fr0 200 I i 1*5 2 29 3!6 4^a DAY OF STUDY 10 11 12 13 1*4 15 16 17 18 19 20 2^ \ DAY OF GESTATION DAY OF LACTATION a. Last value recorded before cohabitaton. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) BODY WEIGHTS Fo GENERATION FEMALE RATS Figure 3 - Groups 5,6,7 I5 1w ,,s yW** y** **8 r . ft GROUP 5 O GROUP 6 GROUP 7 #p<0.05 (from group 1) *p<0.05 (from group 5) **p< 0.0l (from group 5) -V HOVd:8lO_8lF 000073 4^a DAY OF STUDY I i i i I ii i i 1!0 Vi 1^2 1!3 1^ 15 16 17 1*8 l'g 2*0 h DAY OF GESTATION DAY OF LACTATION a. Last value recorded before cohabitaton. APPENDIX B REPORT TABLES 000074 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 1 (PAGE 1): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS I VEHICLE CONTROL 8 0.4 9 0.8 10 11 12 1 1.2 1.6 PRECOHABITATION (DAY 1 OF STUDY TO THE DAY OF COHABITATION): MAXIMUM POSSIBLE INCIDENCE 1176/ 28 840/ 20 840/ 20 840/ 20 840/ 20 1176/ 28 MORTALITY 000000 LOCALIZED ALOPECIA: TOTAL UNDERSIDE LIMBS INCISORS: TOTAL MISSING/BROKEN MISALIGNED 61/ 4 14/ 2 47/ 2 0/ 0 0/ 0 0/ 0 29/ 2 0/ 0 29/ 2 5/ 1 0/ 0 5/ 1 0/ 0 0/ 0 0/ 0 28/ 2 3/ 1 25/ 1 14/ 2 0/ 0 14/ 2 0/ 0 0/ 0 0/ 0 26/ 1 0/ 0 26/ 1 0/ 0 0/ 0 0/ 0 32/ 1 0/ 0 32/ 1 6/ 1 6/ 1 0/ 0 CHROMODACRYORRHEA 0/ 0 8/ 1 19/ 1 0/ 0 0/ 0 0/ 0 FOREPAWS AND LEFT FORELIMB: SCABS 0/ 0 25/ 1 0/ 0 0/ 0 0/ 0 0/ 0 TAIL BENT 40/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 RIGHT FOREPAW: 2ND DIGIT MISSING 16/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 RIGHT FOREPAW: 2ND DIGIT SWOLLEN 13/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. 13 2 1176/ 28 0 26/ 1 0/ 0 26/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 418-018:PAGE B000075 418-018:PAGE B-: 000076 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 1 (PAGE 2): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 12 1 1.2 1.6 PRESUMED GESTATION: MAXIMUM POSSIBLE INCIDENCE 615/ 28 434/ 20 419/ 19 435/ 20 417/ 19 594/ 27 MORTALITY 00000 0 LOCALIZED ALOPECIA: TOTAL LIMBS NECK UNDERSIDE BACK 57/ 3 22/ 1 4/ 1 31/ 2 0/ 0 42/ 2 42/ 2 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 73/ 50/ 0/ 23/ 0/ 5 3 0 2 0 41/ 3 23/ 2 0/ 0 10/ 1 12/ 2** 80/ 5 43/ 2 16/ 2 27/ 2 0/ 0 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 8/ 3 8/ 3 0/ 0 7/ 1 7/ 1 0/ 0 21/ 1 7/ 1 21/ 1 9/ 1 9/ 1 0/ 0 0/ 0 0/ 0 0/ 0 7/ 2 7/ 2 0/ 0 SWOLLEN SNOUT 0/ 0 0/ 0 0/ 0 1/ 1 0/ 0 0/ 0 CHROMODACRYORRHEA 0/ 0 0/ 0 21/ 1 0/ 0 0/ 0 0/ 0 CHROMORHINORRHEA 0/ 0 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 TAIL BENT 22/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 RIGHT FOREPAW: 2ND DIGIT MISSING 22/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 RED PERIVAGINAL SUBSTANCE 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. ** Significantly different from the Group 1 value (p<0.01). 13 2 610/ 28 0 36/ 36/ 5/ 0/ 0/ 3 3 1 0 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 to PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 1 (PAGE 3): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 12 1 1.2 1.6 LACTATION : MAXIMUM POSSIBLE INCIDENCE 95/ 19 100/ 20 85/ 17 93/ 19 90/ 18 76/ 17 LOCALIZED ALOPECIA : TOTAL LIMBS NECK UNDERSIDE BACK IS/ 3 5/ 1 0/ 0 10/ 2 0/ 0 10/ 2 10/ 2 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 17/ 4 13/ 3 0/ 0 4/ 1 0/ 0 13/ 3 9/ 2 0/ 0 4/ 1 8/ 2 15/ 3 10/ 2 1/ 1 5/ 1 0/ 0 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 4/ 1 4/ 1 1/ 1 1/ 1 1/ 1 0/ 0 5/ 1 4/ 1 2/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 CHROMODACRYORRHEA 0/ 0 0/ 0 5/ 1 0/ 0 0/ 0 0/ 0 LACRIMATION 0/ 0 0/ 0 2/ 1 0/ 0 0/ 0 0/ 0 RIGHT FOREPAW: 2ND DIGIT MISSING 5/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 TAIL BENT 5/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 RED PERIVAGINAL SUBSTANCE 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. n /n = T o t a l n u m b e r o f o b s e r v a t i o n s /n u m b e r o f r a t s w i t h o b s e r v a t i o n . 13 2 61/ 19 9/ 3 6/ 3 5/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 -S3DVj:8lO-8Ifr 000077 UJ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 1 (PAGE 4): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 34 1.6 + MEVALONIC ACID 2 + MEVALONIC ACID PRECOHABITATION (DAY 1 OF STUDY TO THE DAY OF COHABITATION): MAXIMUM POSSIBLE INCIDENCE 1176/ 28 1178/ 29 1176/ 28 FOUND DEAD 0 la 0 EXCESS SALIVATION 3/ 2 4/ 3 12/ 7 CHROMORHINORRHEA 2/ 2 0/ 0 3/ 3 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 7/ 2 7/ 2 0/ 0 15/ 4 15/ 4 0/ 0 17/ 2 12/ 2 17/ 1 MOUTH : SCAB 0/ 0 8/ 2 7/ 1 SWOLLEN SNOUT 0/ 0 3/ 1 6/ i TIP OF TAIL MISSING 0/ 0 0/ 0 39/ i PTOSIS 0/ 0 0/ 0 2/ 1 SOFT OR LIQUID FECES 0/ 0 0/ 0 2/ 1 RALES 1/ 1 8/ 1 0/ 0 RED SUBSTANCE IN CAGE 0/ 0 1/ 1 0/ 0 LABORED BREATHING SCANT FECES 0/ 0 0/ 0 1/ 1 1/ 1 0/ 0 0/ 0 LOCALIZED ALOPECIA: UNDERSIDE 8/ 1 0/ 0 0/ 0 RIGHT EAR SWOLLEN 2/ 1 0/ 0 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION, a. Rat 10936 was found dead on day 2 of study. 0000 418-018:PAGE B-4 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 1 (PAGE 5): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 34 1.6 + MEVALONIC ACID 2 + MEVALONIC ACID PRESUMED GESTATION: MAXIMUM POSSIBLE INCIDENCE 609/ 28 616/ 28 609/ 28 MORIBUND SACRIFICED la 0 0 EXCESS SALIVATION 14/ 8##a 10/ 8 14/ 9 PERIORAL SUBSTANCE b CHROMORHINORRHEA 7/ 5##a 3/ 3##a 2/ 2 2/ 2 11/ 7 5/ 2 INCISORS: MISSING/BROKEN PTOSIS LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE NECK 3/ 3 0/ 0 33/ 3 10/ 1 23/ 2 0/ 0 4/ 2 0/ 0 22/ 3 9/ 1 5/ 2 12/ 1 3/ 2 30/ 2 5/ 1 5/ 1 0/ 0 0/ 0 CHROMODACRYORRHEA TIP OF TAIL MISSING 1/ la 0/ 0 0/ 0 0/ 0 2/ 1 22/ 1 DEHYDRATION RALES 0/ 0 0/ 0 0/ 0 0/ 0 5/ 1 5/ 1 EMACIATION SCANT FECES MOUTH : SCAB 0/ 0 0/ 0 3/ 1 0/ 0 0/ 0 4/ 1 2/ 1 1/ 1 0/ 0 SWOLLEN SNOUT 0/ 0 1/ 1 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Dam 10926 was moribund sacrificed on day 12 of gestation,- death was attributed to an intubation accident. b. Descriptions were (or were any combination of) red, dried, slight and moderate, ft# Significantly different from the Group 1 value (p<0.01). 418-018(PAGE B-5 000079 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-G29S.25) TABLE 1 (PAGE 6) : CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID PRESUMED GESTATION: (CONT.) MAXIMUM POSSIBLE INCIDENCE 609/ 28 616/ 28 609/ 28 MORIBUND SACRIFICED la 0 0 RIGHT EAR SWOLLEN RED PERIVAGINAL SUBSTANCE 3/ 1 1/ i 0/ 0 0/ 0 0/ 0 0/ 0 DECREASED MOTOR ACTIVITY 1/ la 0/ 0 0/ 0 SOFT OR LIQUID FECES LACTATION : 1/ la 0/ 0 0/ 0 MAXIMUM POSSIBLE INCIDENCE 90/ 18 76/ 18 52/ 19 PTOSIS 0/ 0 0/ 0 5/ 1 LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE 5/ 1 5/ 1 0/ 0 5/ 1 5/ 1 5/ 1 0/ 0 0/ 0 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Dam 10926 was moribund sacrificed on day 12 of gestation; death was attributed to an intubation accident. 418-018:PAGE B-6 ooooao PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 1 (PAGE 7): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1 . 6 + CHOLESTEROL 7 2 + CHOLESTEROL PRECOHABITATION (DAY 1 OF STUDY TO THE DAY OF COHABITATION): MAXIMUM POSSIBLE INCIDENCE 1150/ 28 1176/ 28 1176/ 28 MORIBUND SACRIFICED la 0 0 LOCALIZED ALOPECIA: TOTAL LIMBS NECK UNDERSIDE INCISORS: TOTAL MISSING/BROKEN MISALIGNED 47/ 6 44/ 5 11/ 2 3/ 1 17/ i 0/ 0 17/ 1 13/ 2 13/ 2 0/ 0 0/ 0 8/ 1 8/ 1 0/ 0 12/ 1 12/ 1 0/ 0 0/ 0 2/ 1 2/ 1 0/ 0 CHROMORHINORRHEA 0/ 0 1/ 1 0/ 0 DECREASED MOTOR ACTIVITY PALE IN APPEARANCE COLD TO TOUCH 1/ la 1/ la 1/ la 0/ n 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 SWOLLEN SNOUT 1/ i 0/ 0 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION, a. Rat 11562 was moribund sacrificed on day 16 of study. T90000 418-018:PAGE B-7 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 1 (PAGE 8): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL, CONTROL 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL PRESUMED GESTATION: MAXIMUM POSSIBLE INCIDENCE MORTALITY 601/ 27 0 604/ 28 0 610/ 28 0 LOCALIZED ALOPECIA: TOTAL NECK LIMBS UNDERSIDE BACK INCISORS: TOTAL MISSING/BROKEN MISALIGNED 77/ 5 22/ 1 59/ 4 14/ 1 0/ 0 32/ 2 0/ 0 32/ 2 52/ 5 0/ 0 41/ 4 28/ 2 14/ 1 17/ 2 16/ 2 7/ 1 54/ 5 0/ 0 40/ 4 22/ 1 2/ i 9/ 2 9/ 2 0/ 0 CHROMODACRYORRHEA 3/ 1 2/ 1 8/ 2 CHROMORHINORRHEA 0/ 0 0/ 0 2/ 2 CHEST OR LEFT FORELIMB: ULCERATION CHEST: SCAB 0/ 0 0/ 0 11/ 1 4/ 1 0/ 0 0/ 0 LACTATION: MAXIMUM POSSIBLE INCIDENCE 85/ 17 86/ 20 65/ 19 LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE BACK 16/ 4 12/ 3 4/ 1 0/ 0 7/ 2 7/ 2 5/ 1 2/ 1 18/ 5 15/ 4 4/ 1 3/ 1 RED PERIVAGINAL SUBSTANCE INCISORS: MISALIGNED 2/ 2 4/ 1 1/ i 4/ 1 0/ 0 0/ 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. 290000 418-018:PAGE B-8 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 1 1.2 12 1.6 RATS EXAMINED a N 28 20 20 20 20 28 MORTALITY N0 0 0 0 0 0 APPEARED NORMAL N 28 20 20 20 20 28 a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at n e c r o p s y . 13 2 28 0 28 80000 6-8 H9VcT8I0-8It7 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 (PAGE 2): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS EXAMINED a N 28 29 2 7b MORTALITY Ni 1 0 MORIBUND SACRIFICED N lc 0 0 FOUND DEAD N0 id 0 APPEARED NORMAL N 26 28 27 ESOPHAGUS: PERFORATION N lc 0 0 LEFT AXILLARY AREA: TISSUE DISCOLORED TAN N lc 0 0 KIDNEYS: BILATERAL, MISSHAPEN N 1 0 0 BILATERAL, PELVIS, SLIGHT DILATION N 1 0 0 BLADDER: WALLS THICK; CONTAINED ONE CALCULUS N 1 0 0 THORACIC CAVITY: CONTAINED DARK RED CLOTTED MATERIAL N 0 Id 0 a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. b. Excludes rat 11554, which did not have necropsy observations recorded. c. Rat 10926 was moribund sacrificed on day 12 of gestation; death was attributed to an intubation accident. d. Rat 10936 was found dead on day 2 of study. *$<*000 418-018:PAGE B-10 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 (PAGE 3): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL RATS EXAMINED a N 28 28 28 MORIBUND SACRIFICED N lb 0 0 APPEARED NORMAL N 27 28 27 SMALL INTESTINES: TWISTED AND CONSTRICTED AT JEJUNUM N lb 0 0 JEJUNUM: CONTAINED DARK RED FLUID N lb 0 0 STOMACH: CONTAINED PUP TISSUE N 0 0 1 a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. b. Rat 11562 was moribund sacrificed on day 16 of study 418-018 :PAGE B000085 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 (PAGE 1) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0 .8 10 11 12 1 1.2 1.6 13 2 CAESAREAN SECTIONED ON DAY 21 OF GESTATION: RATS TESTED N 8 88 TERMINAL BODY WEIGHT MEAN+S.D. 417.5 + 18.3 399.8 + 25.2 409.8 + 31.5 LIVER M E A N + S .D . 14.3 + 0.7 14.9 + 1.9 15.3 + 1.2 LIVER (%) M E A N + S .D . 3.4 + 0.2 DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION: 3.7 0.5 3.7 0.3 RATS TESTED N 19 20 17 18 18 14 6 TERMINAL BODY WEIGHT M E A N + S .D . 318.7 + 28.5 326.7 + 21.6 308.3 + 42.0 309.5 + 19.8 305.6 + 23.2 298.6 + 19.8 283.8 + 7.5* LIVER M E A N + S .D . 13.2 + 1.8 13.8 + 1.5 13.6 + 1.1 13.7 + 1.4 1 4 .5 1.5 13.2 + 1.2 13.1 + 1.0 LIVER (%) M E A N + S .D . 4 .i + 0.5 4.2 + 0.3 4.5 + 0.5 + 4.4 + 0.5 4.8 + 0.3** 4.4 + 0.3 4.6 + 0.3* AL L WEIGHTS WERE RECORDED IN GRAMS (G) . * Significantly different from the Group 1 value (p<0.05). *+ Significantly different from the Group 1 value (p<0.01). RATIOS (%) = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100 . 9S#000 418-018-.PAGE B-12 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 (PAGE 2) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL CAESAREAN SECTIONED ON DAY 21 OF GESTATION: RATS TESTED N 8 TERMINAL BODY WEIGHT M E A N + S .D . 416.6 + 24.6 LIVER M E A N + S .D . 15.3 + 2.1 LIVER (%) M E A N + S .D . 3.7 0.4 DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION: RATS TESTED N 18 TERMINAL BODY WEIGHT M E A N + S .D . 319.5 + 18.5 LIVER M E A N + S .D . 13.3 + 1.2 LIVER (%) MEAN+S.D. 4.2 + 0.3 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID 7 393.7 + 30.4 16.2 + 2.3 4.1 + 0.4* 8 393.1 + 47.3 15.8 + 1.5 4.0 + 0.2 12 3 315.8 + 23.4 297.0 + 14.1 14.8 + 1.2** 13.8 + 1.6 4.7 + 0.3** 4.6 + 0.4* RATIOS (%) = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. 000 418-018:PAGE B-13 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 (PAGE 3) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL CAESAREAN SECTIONED ON DAY 21 OF GESTATION: RATS TESTED N 8 TERMINAL BODY WEIGHT M E A N + S .D . 441.0 + 30.6 LIVER M E A N + S .D . 14 .1 + 1.3 LIVER (%) M E A N + S .D . 3.2 + 0.2 DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION: RATS TESTED N 17 TERMINAL BODY WEIGHT M E A N + S .D . 328.0 + 31.9 LIVER M E A N + S .D . 13.4 + 1.4 LIVER (%) MEAN+S.D. 4.1 + 0.3 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . * Significantly different from the Group 5 value (p<0.05). ** Significantly different from the Group 5 value (p<0.0l). 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL 8 403.0 + 38.3* 14 .3 + 0.8 3.6 + 0.4* 8 387.5 + 31.3** 15.3 + 0.7* 4.0 + 0.3** 13 7 303.9 + 19.0* 280.8 + 15.6** 14 .3 + 1.7 12.6 + 0.9 4.7 + 0.5** 4.5 + 0.2* RATIOS (%) = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) x 100. 418-018:PAGE B-14 000088 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25! TABLE 4 (PAGE 1): BODY WEIGHTS - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0 .4 RATS TESTED N 28 20 BODY WEIGHT (G) DAY 1 M E A N + S .D . 215.4 + 10.8 215.0 + 12.7 DAY 8 M E A N + S .D . 229.8 + 12.1 236.2 + 11.0 DAY 15 DAY 22 DAY 29 M E A N + S .D . M E A N + S .D . 238.5 14 .8 247.9 + 17.6 M E A N + S .D . 255.3 + 17.5 247.5 + 13.8 257.4 t 12.9 262.4 + 15.0 DAY 36 M E A N + S .D . 262.6 + 18.5 271.4 + 16.3 DAY 42a MEAN+S.D. 265.6 + 21.0 276.0 + 16.8 DAY = DAY OF STUDY a. Last value recorded before cohabitation. 9 0 .8 20 215.2 + 11.2 233.0 + 11.0 244.5 + 12.2 252.3 + 15.1 256.8 + 17.3 264.4 + 17.9 266.0 + 19.5 10 1 20 214 .2 + 10.8 231.9 + 14.3 242.0 + 14.9 252.8 + 15.6 260.0 + 15.9 267.0 + 18.7 +269.2 21.2 11 1.2 20 214 .8 + 11.3 233.1 + 12.9 242.2 + 14 .1 251.2 + 14.1 257.6 + 14 .1 264.4 + 14.4 264.6 + 16.7 12 1 .6 28 215.6 + 10.0 235.1 + 11.7 243.5 + 12.6 250.5 + 15.2 256.1 + 15.6 260.2 + 15.2 260.2 + 15.5 13 2 28 215.9 + 10.0 231.3 + 8.9 239.5 + 12.4 246.6 + 14.8 249.4 + 14.6 253.6 + 14.6 255.1 + 17.5 ci-aHDVd:8lO-8It> 000089 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 4 (PAGE 2): BODY WEIGHTS - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 34 1.6 + MEVALONIC ACID RATS TESTED N 28 29 BODY WEIGHT (G) DAY 1 M E A N + S .D . 216.2 + 11.3 215.6 + 12.1 DAY 8 DAY 15 DAY 22 DAY 29 DAY 36 DAY 42b M E A N + S .D . M E A N + S .D . M E A N + S .D . M E A N + S .D . M E A N + S .D . M E A N + S .D . 229.6 + 11.0 240.2 + 14.1 249.1 + 13.6 257.4 + 14.6 265.4 + 17.1 267.6 + 17.4 231.6 + 1 3 . 3 [ 28] a 240.5 + 17.5 [ 28] a 249.5 + 18.7 [ 28] a 253.5 + 19.7 [ 28] a 258.1 + 19.5 [ 28] a 261.3 + 2 3 . 1 E 28] a DAY = DAY OF STUDY ( ] = NUMBER OF VALUES AVERAGED a. Excludes values for rat 10936, which was found dead on day 2 of study. b. Last value recorded before cohabitation. 2 + MEVALONIC ACID 28 215.6 + 11.6 234.2 + 11.5 242.9 + 11.8 252.6 + 13.4 258.2 + 14.2 263.1 + 15.6 266.4 + 16.9 418-018:PAGE B-16 000090 PROTOCOL 418-018; ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 4 (PAGE 3); BODY WEIGHTS - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL RATS TESTED N 28 28 BODY WEIGHT (G) DAY 1 MEAN+S .D . 214.8 + 10.1 215.7 + 10.5 DAY 8 M E A N + S .D . 232.8 + 11.4 231.5 + 10.8 DAY 15 M E A N + S .D . 242.8 + 14.3 240.8 + 12.4 DAY 22 DAY 29 DAY 36 DAY 42b M E A N + S .D . MEAN+S.D. M E A N + S .D . M E A N + S .D . 254.2 + 1 5 . 4 [ 27] a 264.5 + 17.9 [ 27) a 272.1 + 17.1 [ 27] a 275.5 + 19.7 [ 27] a 248.8 + 14.5 253.1 + 13.8** 257.4 + 15.7** 257.6 + 16.4** DAY = DAY OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Excludes values for rat 11562, which was moribund sacrificed on day 16 of study, b. Last value recorded before cohabitation. ** Significantly different from the Group 5 value (pcO.Ol). 7 2 + CHOLESTEROL 28 215.4 + 10.9 233.0 + 10.8 242.5 + 12.3 250.2 + 12.3 252.5 + 14.3** 255.5 + 15.0** 257.9 + 14.9** 418-018-.PAGE B-17 000091 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 5 (PAGE 1): BODY WEIGHT CHANGES - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0 .4 9 0.8 RATS TESTED N 28 20 20 BODY WEIGHT CHANGE (G) DAYS 1 - 8 DAYS 8 - 15 DAYS IS - 22 M E A N + S .D . + 14 .5 + 8.6 M E A N + S .D . +8.7 + 5.2 MEAN+S.D. +9.4 + 6.9 DAYS 22 - 29 M E A N + S .D . +7.4 + 5.9 DAYS 29 - 36 M E A N + S .D . +7.3 + 4.7 DAYS 36 - 42a M E A N + S .D . +3.0 + 8.4 +DAYS 1 - 42a MEAN+S.D. +50.2 17.3 +21.2 + 6.6** +17.7 + 6.4 +11.2 + 7.5 +11.6 + 5.2 +10.0 + 6.8 +7.8 + 4.9 +4.9 + 5.9 +4.6 + 6.8 + 9.0 + 5.7 +7.6 + 5.2 +4.7 + 7.6 +1.6 + 9.1 ++61.0 15.0* +50.8 + 15.5 DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. * Significantly different from the Group 1 value ** Significantly different from the Group 1 value (P5O . 05). (p<0.01). 10 1 20 +17.7 + 8.7 +10.1 + 6.1 +10.8 + 5.1 +7.2 + 5.4 +7.0 + 4 .4 +2.3 + 8.2 ++55.0 17.9 11 1.2 20 + 18.3 + 9.1 +9.0 + 6.4 +9.0 + 5.0 +6.5 + 6.3 +6.7 + 4.0 +0.2 + 8.5 +49.8 + 14 .1 12 13 1.6 2 28 28 +19.5 + 6.9* + 15.4 + 6.6 +8.4 + 5.5 + 8.2 + 6.6 +7.0 + 4.2 +7.1 + 6.6 +5.6 + 5.0 +2.8 + 4.9 +4.2 + 4.9* -0.1 + 6.0 +44.5 + 10.4 +4.2 + 4 .2* ++ 1.5 6.8 +39.2 + 16.4* 418-018:PAGE B-18 000092 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 5 (PAGE 2): BODY WEIGHT CHANGES - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP1 DOSAGE MG/KG; p f o s 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID RATS TESTED N 28 29 INCLUDED IN ANALYSES N 28 28a BODY WEIGHT CHANGE (G) DAYS 1 - 8 M E A N + S .D . + 13.4 + 8.0 +15.9 + 7.6 DAYS 8 - 15 M E A N + S .D . + 10.6 + 5.3 +8.8 + 6.6 DAYS 15 - 22 M E A N + S .D . +8.9 + 5.5 + 9.0 + 5.4 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 42b DAYS 1 - 42b M E A N + S .D . M E A N + S .D . M E A N + S .D . MEAN+S.D. +8.3 + 6.5 ++8.0 5.9 ++2.1 6.9 ++ 51.4 13.7 +4.1 + 4.7** ++4.5 4.2** ++ 3.2 6.5 ++45.6 16.6 DAYS = DAYS OF STUDY a. Excludes values for rat 10936, which was found dead on day 2 of study. b. Last value recorded before cohabitation. * Significantly different from the Group 2 value (p<0.05), ** Significantly different from the Group 2 value (p<0.01). 4 2 + MEVALONIC ACID 28 28 +18.6 + 5.2" +8.7 + 4.3 +9.7 + 5.2 +5.6 + 4 .8 +4.8 + 3.7* +3.3 + 5.0 +50.8 + 10.6 418-018:PAGE B-19 000093 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 5 (PAGE 3): BODY WEIGHT CHANGES - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS CHOLESTEROL CONTROL 1.6 + CHOLESTEROL RATS TESTED N 28 28 BODY WEIGHT CHANGE (G) DAYS 1 * 8 M E A N + S .D. +18.0 + 7.5 +15.8 + 5.8 DAYS 8 - 15 MEAN+S.D. +10.0 + 6.9 +9.3 + 5.3 DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 42b DAYS 1 - 42b M E A N + S .D . M E A N + S .D . MEAN+S.D. M E A N + S .D . M E A N + S .D . +10,7 + 4.8 l 27] a +10.3 + 7.0 [ 27] a +7.7 + 7.8 t 27] a +3.4 + 7.4 t 27] a +60.7 + 16.8# 1 27] a +8.1 + 6.1 +4 .3 + 4 .4** +4.3 + 5.6* +0.1 + 3.4 +41.9 + 11.0** DAYS = DAYS OF STUDY ( 1 = NUMBER OF VALUES AVERAGED a. Excludes values for rat 11562, which was moribund sacrificed on day 16 of s t u d y . b. Last value recorded before cohabitation, (t Significantly different from the Group 1 value (p<0 .05) * Significantly different from the Group 5 value (p<0 .05) ** Significantly different from the Group 5 value (p<0 .01) 2 + CHOLESTEROL 28 +17.6 + 7.2 +9.5 + 6.2 +7.7 + 5.2 +2.2 + 5.6** +3.0 + 4.6** +2.4 + 6.0 +4 2.5 + 14.0** 418-018:PAGE B-20 000094 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 6 (PAGE 1): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE: CONTROL 8 0 .4 9 0..8 RATS TESTED N 28 20 20 PREGNANT N 27 20 17 MATERNAL BODY WEIGHT (G) DAY 0 M E A N + S .D . 275.0 + 21.6 281.6 + 17.0 274.0 + 22.1 DAY 1 MEAN+S.D. 283.3 + 20.4 291.2 + 18.5 281.1 + 23 .3 DAY 2 M E A N + S .D . 287.9 + 20.9 294.6 + 19.7 284.4 + 23.2 DAY 3 MEAN+S.D. 289.0 + 20.2 296.2 + 18.6 284.7 + 22.6 DAY 4 MEAN+S.D. 291.7 + 19.5 299.6 + 19.7 286.6 + 22.9 DAY 5 M E A N + S .D . 293.6 + 19.9 304.0 + 19.8 290.4 + 23.7 DAY 6 M E A N + S .D . 295.4 + 20.2 306.2 + 20.1 289.8 + 22.0 DAY 7 M E A N + S .D . 298.9 + 20.0 308.4 + 19.2 292.5 + 21.7 DAY 8 M E A N + S .D . 299.6 + 21.8 311.8 + 19.1 295.2 + 21.8 DAY 9 MEAN+S.D. 304.0 + 21.9 314.8 + 19.3 289.9 + 22.3 DAY 10 MEAN+S.D. 308.4 + 21.9 318.7 + 19.5 303.5 + 23.0 DAY = DAY OF GESTATION * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 10 1 20 19 274,2 + 19.9 282.6 + 21.1 286.6 + 22.0 287.6 + 19.8 288.2 + 18.1 291.2 + 18.6 292 .1 + 18.5 294 .3 + 19.9 296.8 + 16.5 300.2 + 16.9 303.9 + 17.0 11 1.2 20 18 268.2 + 18.6 275.4 + 17.5 277.6 + 17.8 277.5 + 19.4 278.6 + 18.6 281.3 + 16.8 283 .6 + 18.0 286.1 + 19.2 289.5 + 19.9 291.4 + 19.3 295.3 + 20.3 12 13 1.6 2 28 28 25 27 267.8 + 15.8 259.7 + 19.0* 272.6 + 16.8 265.7 + 19.9** 275.0 + 16.2* 267.0 + 17.9** 275.4 + 16.1* 268.0 + 17.1** 276.5 + 16.5* 269.4 + 17.0** 277.9 + 17.0** 271.8 + 17.0** 280.2 + 16.9* 272.9 + 17.1** 283.6 + 16.6* 276.5 + 17.6** 286.1 + 15.9* 278.7 + 18.3** 288.4 + 19.0* 281.5 + 17.4** 293.1 + 17.8* 286.2 + 17.8** 418-018:PAGE B-2 000095 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 6 (PAGE 2): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED N 1 VEHICLE CONTROL 28 8 0 .4 20 9 0 .8 20 PREGNANT N 27 20 17 MATERNAL BODY WEIGHT (G) DAY 11 M E A N + S .D . 313.8 + 21.1 324.2 + 19.5 308.6 + 23.2 DAY 12 M E A N + S .D . 318.2 + 22.3 329.8 + 19.2 312.9 + 24.7 DAY 13 M E A N + S .D . 322.8 + 22.4 332.0 + 20.2 317.2 + 25.4 DAY 14 MEAN+S.D. 326.8 + 22.6 337.6 + 21.2 323.4 + 27.3 DAY 15 M E A N + S .D . 333.9 + 23.0 347.1 + 20.9 332.4 + 25.3 DAY 16 M E A N + S .D . 343.1 + 24.1 357.0 + 22.6 343.3 + 26.0 DAY 17 M E A N + S .D . 356.0 + 26.6 370.4 + 23.0 357.0 + 26.4 DAY 18 M E A N + S .D . 370.1 + 28.6 383.6 + 23.9 371.2 + 26.1 DAY 19 MEAN+S.D. 381.9 + 29.9 395.8 + 24.4 384.6 + 28.0 DAY 20 M E A N + S .D . 396.2 + 31.3 410.3 + 26.0 395.6 + 26.2 DAY 21 M E A N + S .D . 410.6 + 31.1 421.2 + 28.9 ( 18]!a 408.5 + 25.3 [ 12 ]a DAY = DAY OF GESTATION I ] = NUMBER OF VALUES AVERAGED a. Excludes values for dams that delivered. * Significantly different from the Group 1 value ** Significantly different from the Group 1 value (p<0.05). (pcO.Ol). 10 1 20 19 310.0 + 17.9 312.4 + 18.5 318.3 + 17.2 326.6 + 17.9 333.1 + 18.3 344.8 + 16.8 357.9 + 18.1 373.4 + 18.7 386.7 + 17.1 400.2 + 20.1 408.6 2 2 '2 ( 12] a ii 12 1.2 1 .6 20 28 18 25 13 2 28 27 302.2 + 19.9 296.4 + 18.4** 291.2 + 19.4** 306.8 + 22.5 299.1 + 19.2*+ 294.4 + 18.8** 309.3 + 19.7 304.3 + 19.4** 299.2 + 18.4** 316.2 + 19.4 310.4 + 18.2* 302.9 + 19.0** 322.4 + 21.2 319.1 + 19.0* 313.3 + 19.4** 333.3 + 21.3 328.7 + 19.6* 323.4 + 19.3+* 347.9 + 22.5 343.4 + 20.6 337.3 + 20.0** 362.7 + 23 .7 358.5 + 20.3 352.1 + 21.0* 373.2 + 25.3 373.0 + 21 .8 365.6 + 21.5* 387.1 + 27.0 385.1 + 25.0 380.0 + 23.3 394.5 + 31.7 [ 15]a 398.3 + 26.1 [ 20) a 397.5 + 30.2 E 21] a 960000 418-018:PAGE B-22 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 6 (PAGE 3): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL RATS TESTED N 28 PREGNANT N 27 MATERNAL BODY WEIGHT (G) DAY 0 M E A N + S .D . 275.3 + 19.0 DAY 1 M E A N + S .D . 283.0 + 18.8 DAY 2 M E A N + S .D . 286.3 + 18.9 DAY 3 M E A N + S .D . 287.0 + 18.9 DAY 4 M E A N + S .D . 289.0 + 18.9 DAY 5 DAY 6 DAY 7 DAY 8 DAY 9 DAY 10 M E A N + S .D . M E A N + S .D . MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 290.8 + 19.3 +292.9 19.0 +296.0 17.9 +299.0 18.2 +302.6 19.4 +305.9 19.0 DAY = DAY OF GESTATION a. Excludes rat 10936, which was found dead prior to gestation. * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). 34 1.6 + MEVALONIC ACID 28a 26 266.6 + 22.9 271.5 + 22.3 273.4 + 22.4 273.8 + 23 .1 275.6 + 22.2* 275.9 + 22.1** +277.9 23.3** +280.0 22.3** +282.3 22.9** +285.6 23.1** +289.5 23.4** 2 + MEVALONIC ACID 28 27 272.0 + 16.9 275.7 + 17.5 278.1 + 18.5 278.6 + 19.0 279.1 + 18.4 281.3 + 18.5 282.4 + 18.9 284.7 + 19.9* 286.6 + 18.9* 290.0 + 20.5 + 294.6 + 19.7* 418-018:PAGE B-23 000097 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-S295.25) TABLE 6 (PAGE 4): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 34 1.6 + MEVALONIC ACID 2 + MEVALONIC ACID RATS TESTED N 28 28a 28 PREGNANT N 27 26 27 MATERNAL BODY WEIGHT (G) DAY 11 M E A N + S .D . 311.7 + 18.5 294.4 + 24.9** 300.7 20.5 DAY 12 M E A N + S .D . 313.5 + 18.3 299.0 + 25.2* 303.3 + 21.0 DAY 13 DAY 14 DAY 15 DAY 16 DAY 17 DAY 18 DAY 19 DAY 20 DAY 21 M E A N + S .D . M E A N + S .D . MEAN+S.D. M E A N + S .D . M E A N + S .D . M E A N + S .D . M E A N + S .D . MEAN+S .D . M E A N + S .D . 318.3 + 18.6 [ 26] b 322.8 + 18.8 1 26] b 330.6 + 18.8 [ 26] b 339.4 + 18.8 t 26] b 351.0 + 20.7 [ 26] b 363.4 + 20.8 26] b 379.4 + 24.9 1 26]b 390.9 + 24.7 [ 26] b 403.5 + 24.8 [ 25]b,c 302.9 + 25.9* 308.4 + 26.9 315.9 + 25.8 325.7 + 27.2 340.5 + 27.6 356.4 + 28.9 368.9 + 29.9 382.7 + 32.6 398.2 + 34.1 ( 25] c 309.1 + 20.5 312.2 + 21.6 319.4 + 24.0 329.9 + 25.6 344.2 + 25.8 359.8 + 27.1 372.4 + 26.8 385.3 + 28.4 403.4 + 35.1 [ 19] c DAY = DAY OF GESTATION ( ) = NUMBER OF VALUES AVERAGED a. Excludes rat 10936, which was found dead prior to gestation. b. Excludes values for dam 10926, which was moribund sacrificed on day 12 of gestation accident). c. Excludes values for dams that delivered. * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). (death was attributed to an intubation 418-018:PAGE B-24 000098 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 6 (PAGE 5): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 67 1.6 + CHOLESTEROL RATS TESTED N 27a 28 PREGNANT N 25 28 MATERNAL BODY WEIGHT (G) DAY 0 M E A N + S .D . 284.8 + 22.3 265.1 + 17.9** DAY 1 MEAN+S.D. 292.6 + 22.7 268.2 + 19.4** DAY 2 MEAN+S.D. 297.6 + 21.7 271.5 + 18.7** DAY 3 M E A N + S .D . 299.4 + 2 2 . 6 272.6 + 18.5** DAY 4 M E A N + S .D . 302.3 + 21.4 274.2 + 18.3** DAY 5 M E A N + S .D . 305.7 + 22.4# 277.2 + 19.5** DAY 6 MEAN+S .D . 306.6 + 25.2 279.6 + 19.5** DAY 7 M E A N + S .D . 309.8 + 2 4 . 0 281.5 + 20.5** DAY 8 MEAN+S .D . 315.1 + 25.0ff 283.8 + 21.0** DAY 9 M E A N + S .D . 317.7 + 24.6tt 287.8 + 19.6** DAY 10 MEAN+S.D. 323.6 + 24.4# 292.8 + 20.5** DAY = DAY OF GESTATION a. Excludes rat 11562, which was moribund sacrificed prior to gestation. ft Significantly different from the Group 1 value (pfO.05). ** Significantly different from the Group 5 value (p<0.01). 2 + CHOLESTEROL 28 27 264.0 + 17.9** 267.1 + 18.3** 270.4 + 17.7** 270.1 + 15.4** 271.4 + 17.3** 273.4 + 16.7** 276.0 + 16.4** 278.4 + 17.7** 282.2 + 17.5** 284.9 + 17.2** 288.8 + 19.2** 418-018:PAGE B-25 000099 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 6 (PAGE 6): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL, CONTROL 6 1 . 6 + CHOLESTEROL RATS TESTED N 27a 28 PREGNANT N 25 28 MATERNAL BODY WEIGHT (G) DAY ii M E A N + S .D . 329.0 + 24 .2# 295.8 + 21.4** DAY 12 M E A N + S .D . 332.9 + 25.1# 300.8 + 22.6** DAY 13 M E A N + S .D . 335.7 + 26.0# 305.6 + 22.1*+ DAY 14 M E A N + S .D . 340.7 + 25.1# 310.1 + 23.8** DAY 15 MEAN+S .D . 347.6 + 24.4# 318.4 + 24.9** DAY 16 DAY 17 DAY 18 DAY 19 DAY 20 DAY 21 M E A N + S .D . MEAN+S.D. M E A N + S .D . M E A N + S .D . M E A N + S .D . MEAN+S.D. 357.3 + 26.8# +370.2 26.2# +383.2 27.0 +397.1 28.6 +410.3 29.6 +428.0 31 .1# [ 24] b 329.0 + 26.4** +342.4 26.8** +357.2 29.2** +370.1 31.7** +383.6 32.6** +392.0 31.6** [ 23]lb DAY = DAY OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rat 11562, which was moribund sacrificed prior to gestation. b. Excludes values for dams that delivered. # Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 28 27 292.8 + 18.7** 296.3 + 20.4** 300.8 + 21.5** 305.4 + 21.0** 314.4 + 21.9** 325.4 + 23 .2** 339.7 + 24.1** 354.8 + 24.9** 369.3 + 26.2** 380.9 + 28.1** 394.5 + 29.0** [ 19] b 418-018:PAGE B-26 000100 PROTOCOL 4IB-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 7 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 il 12 1 1.2 1.6 13 2 RATS TESTED N 28 20 20 20 20 28 28 PREGNANT N 27 20 17 19 18 25 27 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 MEAN+S.D. +23.8 + 7.0 +26.B + 7.5 +18.5 + 8.6* +20.0 + 7.0 +17.9 + 6.2* +15.8 + 5.3* * +16.8 + 6.0** DAYS 7 - 14 MEAN+S.D. +27.9 + 6.1 +29.2 + 8.7 +30.9 + 8.5 +32.3 + 7.5 +30.1 + 6.9 +26.7 + 6.4 +26.4 + 6.5 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. +83.8 + 13.3 MEAN+S.D.+135.5 + 15.6 +84.3 + 13.6 ( 18] a +139.7 + 17.5 I 18] a +86.2 + 7.6 [ 12] a +133.4 + 13.7 ! 12 ]a +78.4 + 25.6 [ 12] a +132.6 + 29.6 [ 12 ]a +79.9 + 16.6 [ 15] a +128.3 + 23 .5 ( 15] a +87.2 + 15.3 [ 20] a +129.6 + 16.7 [ 20] a +96.8 + 20.0* [ 21] a +138.9 + 20.3 [ 21 ]a DAYS = DAYS OF GESTATION [ I = NUMBER OF VALUES AVERAGED a. Excludes values for dams that delivered. * Significantly different from the Group 1 value ** Significantly different from the Group 1 value (p<0.05). (p<0.01). TOTOOO 418-018:PAGE B-27 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 7 (PAGE 2): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS TESTED N 28 28a 28 PREGNANT N 27 26 27 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 M E A N + S .D . +20.7 + 6.1 +13.4 + 7.4** +12.7 + 7.8** DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. +27.3 + 7.2 ( 26) b +81.8 + 14.5 [ 25] b, c +130.1 + 17.8 [ 25]b,c +28.4 + 6.5 +90.4 + 15.5 [ 25] c +132.2 + 1 8 . 3 [ 25] c +27.6 + 8.2 +88.8 + 23.8 [ 19] C +130.1 + 30.1 ( 19] c DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rat 10936, which was found dead prior to gestation. b. Excludes values for dam 10926, which was moribund sacrificed on day 12 of gestation accident). c. Excludes values for dams that delivered. ** Significantly different from the Group 2 value (p<0.01). (death was attributed to an intubation 418-018:PAGE B-28 000102 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 7 (PAGE 3); MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL RATS TESTED N 27a 28 PREGNANT N 25 28 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 M E A N + S .D . +25.0 + 10.4 +16.4 + 8.9** DAYS 7 - 14 M E A N + S .D . +30. 9 + 6.9 +28.6 + 8.2 DAYS 1 4 - 2 1 DAYS 0 - 21 MEAN+S. D. MEAN+S.D. +86.5 + 1 4 . 9 ( 241b +142.8 + 1 9 . 8 l 24] b +86.4 + 18.5 f 23] b +129.7 + 22.8* 1 231b DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rat 11562, which was moribund sacrificed prior to gestation. b. Excludes values for dams that delivered. * Significantly different from the Group 5 value (p<0.05). ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 28 27 + 14 .4 + 6.1** +26.9 + 7.0 +89.6 + 18.7 1 19] b +128.5 + 15.4* [ 19] b COTOPO 418-018:PAGE B-29 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 8 (PAGE 1): BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 1 1.2 12 1.6 RATS TESTED N 19 20 20 20 20 20 PREGNANT N 19 20 17 19 IB 17 DELIVERED A LITTER N 19 20 17 19 18 17 BODY WEIGHT (G) DAY 1 M E A N + S .D . 301.3 + 26.0 311.0 + 19.9 304.0 + 24.6 304.4 + 15.5 298.1 + 20.1 298.2 + 20.0 DAY 5 MEAN+S.D, 318.7 + 28.5 326.7 + 21.6 308.3 + 42.0 309.5 + 19.8 ( 18] a 305.6 + 23.2 298.6 + 19.8 [ 14] a DAY = DAY OF LACTATION ( ] = NUMBER OF VALUES AVERAGED a. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. * Significantly different from the Group 1 value (p<0.05). 13 2 20 19 19 291.0 + 21.2 283.8 + 7.5* [ 6] a 418-018:PAGE B000104 LO.J PROTOCOL 41B-018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 8 (PAGE 2): BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS TESTED N 19a 20a 20 PREGNANT N 18 18 19 DELIVERED A LITTER N 18 18 19 INCLUDED IN ANALYSES N 18 18 18b BODY WEIGHT (G) DAY 1 M E A N + S .D . 299.3 + 2 4 . 8 301.4 + 2 8 . 6 300.8 + 20.0 DAY 5 MEAN+S.D. 319.5 + 18.5 315.8 + 23.4 I 12] b 297.0 + 1 4 . 1 [ 3]b DAY = DAY OF LACTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rats that died or were moribund sacrificed prior to lactation. b. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. 418-018 :PAGE B-: 000105 U> PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 8 (PAGE 3): BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1 .6 + CHOLESTEROL 7 2 + CHOLESTEROL RATS TESTED N 19a 20 20 PREGNANT N 17 20 19 DELIVERED A LITTER N 17 20 19 BODY WEIGHT (G) DAY 1 M E A N + S .D . 312.4 + 2 4 . 3 294.8 + 24.8* 295.4 + 22.6* DAY 5 M E A N + S .D . 328.0 + 3 1 . 9 303.9 + 19.0* [ 13 Jb 280.8 + 15.6** ( 7]b DAY = DAY OF LACTATION ( ] = NUMBER OF VALUES AVERAGED a. Excludes rat 11562, which was moribund sacrificed prior to lactation, b. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. * Significantly different from the Group 5 value (pcO.05). ** Significantly different from the Group S value (p<0.01). 90T000 -e3 D V d :8lO'8It' Lto.J PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 9 (PAGE 1): BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL a 0.4 9 0.8 10 11 12 1 1.2 1.6 13 2 RATS TESTED N 19 20 20 20 20 20 20 PREGNANT N 19 20 17 19 18 17 19 DELIVERED A LITTER N 19 20 17 19 18 17 19 INCLUDED IN ANALYSES N 19 20 17 18a 18 14a 6a BODY WEIGHT CHANGE (G) DAYS 1 - 5 MEAN+S.D . +17.4 + 11.1 +15.7 + 15 .4 + 4 . 3 + 25 .8* +4.0 + 10.5* +7.5 + 11.6 +1.1 + 12.7** -3.5 + 16.8* DAYS = DAYS OF LACTATION a. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. * Significantly different from the Group I value (p<0.05). ** Significantly different from the Group 1 value (pcO.Ol). PROTOCOL 418-018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 9 (PAGE 2): BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS TESTED N 19a 20a 20 PREGNANT N 18 18 19 DELIVERED A LITTER N 18 18 19 INCLUDED IN ANALYSES N 18 12b 3b BODY WEIGHT CHANGE (G) DAYS 1 - 5 MEAN+S.D. +20.2 + 15.3 +7.7 + 15.5* DAYS = DAYS OF LACTATION a. Excludes rats that died or were moribund sacrificed prior to lactation. b. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. * Significantly different from the Group 2 value (p<0.05). +1.7 _+ 11.2 418-018:PAGE B- 000108 LJ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 9 (PAGE 3): BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL RATS TESTED N 19a 20 20 PREGNANT N 17 20 19 DELIVERED A LITTER N 17 20 19 INCLUDED IN ANALYSES N 17 13b 7b BODY WEIGHT CHANGE (G) DAYS 1 - 5 MEAN+S.D. +15.7 + 19.8 +7.6 + 18.4 -5.0 + 9.8* DAYS = DAYS OF LACTATION a. Excludes rat 11562, which was moribund sacrificed prior to lactation. b. Excludes values for dams that were sacrificed due to no surviving pups * Significantly different from the Group 5 value (p<0.05). prior to day 5 of lactation. 418-018:PAGE B-35 000109 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 10 (PAGE 1) : ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG /KG PFOS 1 VEHICLE CONTROL 8 0.4 RATS TESTED N 28 20 FEED CONSUMPTION (G/DAY) DAYS 1 - 8 DAYS 8 - 15 M E A N + S .D . 19.6 + 3.0 ( 26] a M E A N + S .D . 19.0 + 1.8 19.6 + 1.9 [ 19] a 19.0 + 3.6 DAYS 15 - 22 DAYS 22 - 29 M E A N + S .D , 18.9 + 2.3 MEAN+S .D . 18.8 + 2.0 19.4 + 1.8 [ 19] a 19.4 + 2.0 DAYS 29 - 36 M E A N + S .D , 18.8 + 2.1 DAYS 36 - 42b M E A N + S .D 18.3 + 2.1 20.2 + 2.2* [ 17] a 19.4 + 2.1 DAYS 1 - 42b M E A N + S .D ,. 18.9 + 1.8 19.6 + 1.8 9 0.8 20 1 9 . 1 + 1.9 [ 19] a 18.9 + 1.5 ( 19] a 19.2 + 1.6 [ 18] a 19.0 + 2.5 [ 19] a 18.3 + 2.1 [ 18} a 18.2 + 2.6 19.0 + 1.8 DAYS = DAYS OF STUDY ( ] = NUMBER OF VALUES AVERAGED a. Excludes values that were associated with spillage. b. Last value recorded before cohabitation. * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 10 11 1 1.2 20 20 18.9 + 2.2 19.4 + 2.9 [ 19] a 20.4 + 2.2* 19.8 + 1.8 19.7 + 2.2 ( 19] a 18.3 + 2.0 [ 19] a 19.4 + 1.8 [ 19] a 1 9 . 2 + 1.5 18.7 + 1.8 19.1 + 1.4 [ 17] a 1 8 . 8 + 1.7 ( 19] a 18.3 + 1.7 1 19] a 17.4 + 2.2 ( 19] a 18.6 + 1.3 i 19} a 12 13 1.6 2 28 28 19.9 + 2.0 18.9 + 1.9 19.1 + 2.1 [ 27] a 18.5 + 1.6 [ 27] a 18 3 2.2 17.0 + 1.6* 18.7 + 1.6 19.0 + 1.8 [ 27] a 18.0 + 1.5 18.2 + 1.6 [ 27] a 17.6 + 1.7* 17.0 + 1.5** [ 26] a 16.3 + 1.6** [ 27] a 1 7 . 8 + 1.3* [ 27] a 418-018:PAGE B- 000110 LC.bJ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER; T-6295.2S) TABLE 10 (PAGE 2): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED INCLUDED IN ANALYSES N N 2 MEVALONIC ACID CONTROL 28 28 3 1.6 + MEVALONIC ACID 29 28a FEED CONSUMPTION (G/DAY) DAYS 1 - 8 MEAN+S.D. DAYS 8 - 15 MEAN+S.D. DAYS 15 - 22 MEAN+S.D. DAYS 22 - 29 MEAN+S.D. DAYS 29 - 36 MEAN+S.D. DAYS 36 - 42c MEAN+S.D. DAYS 1 - 42c MEAN+S.D. 19.9 + 1.4 ( 26) b 2 0 . 1 + 2.2 20.1 + 2.2 [ 27] b 20.0 + 1.9 19.4 + 2.2 [ 26) b 1 8 . 8 + 2.1 1 9 . 8 + 1.7 19.2 + 1.9 19.0 + 3.4 18.8 + 2.5 ( 27] b 18.0 + 2.4** [ 27] b 18.4 + 2.2 17.5 + 2.3* [ 27]b 18.5 + 2.0** [ 27] b DAYS = DAYS OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Excludes values for rat 10936, which was found dead on day 2 of study. b. Excludes values that were not recorded or were associated with spillage. c. Last value recorded before cohabitation, * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). 4 2 + MEVALONIC ACID 28 28 19.5 + 1.4 [ 27] b 18.8 + 2.1 19.0 + 1.9 [ 27] b 18.4 + 1.7** [ 26] b 18.3 + 2.5 ( 26] b 17.4 + 2.3* 18.7 + 1.5* TTTOOO 418-018:PAGE B- L-1.J PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 10 (PAGE 3): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG: PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL RATS TESTED N 28 28 FEED CONSUMPTION (G/DAY) DAYS 1 - 8 M E A N + S .D . 19.1 + 1.5 18.7 + 1.6 DAYS 8 - 15 DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 42c DAYS 1 - 42c M E A N + S .D . M E A N + S .D . M E A N + S .D . M E A N + S .D . M E A N + S .D . M E A N + S .D . 19.2 + 1.8 [ 27] a 20.0 + 2.0 ( 27] b 19.8 + 2.0 [ 27] b 20.9 + 2.5## [ 25] a ,b 19.7 + 2.1# [ 27] b 1 9 . 7 + 1.7 ( 27] b 18.6 + 1.3 18.8 + 2.1 17.9 + 1.5** 18.3 + 1.4** I 23Ja 17.3 + 1.5** 18.3 + 1.2** DAYS = DAYS OF STUDY ( ] = NUMBER OF VALUES AVERAGED a . Excludes values that were associated with spillage. b. Excludes values for rat 11562, which was moribund sacrificed on day 16 of study. c. Last value recorded before cohabitation. # different from the Group 1 value (p<0.05) ## different from the Group 1 value (pcO.Ol) * * different from the Group 5 value (p<0.01) 7 2 + CHOLESTEROL 28 19.0 + 1.5 18.9 + 1.9 18.9 + 2.1 I 27] a 3.7.7 + 1.9** 18.2 + 2.4** 1 25] a 17.3 + 1.6** 18.4 + 1.6** 418-018:PAGE B- 000112 L .2 OO PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 11 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS i VEHICLE CONTROL 8 0.4 RATS TESTED N 26 20 FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 DAYS 8 - 15 MEAN+S.D. MEAN+S. D. 88.2 + 14.0 ( 26) a 80.9 + 4 .9 87.0 + 8.3 [ 19) a 78.4 + 14.4 DAYS 15 - 22 DAYS 22 - 29 M E A N + S .D . 77.7 + 6.3 M E A N + S .D . 74.9 + 5.3 7 7 . 1 + 6.8 [ 19] a 7 4 . 6 + 6.2 DAYS 29 - 36 M E A N + S .D . 72.6 + 6.4 DAYS 36 - 42b M E A N + S .D . 69.1 + 5.4 7 6 . 6 + 6.9 ( 17] a 70.7 + 5.3 DAYS 1 - 42b M E A N + S .D . 77.2 + 4.6 77.6 + 5.8 9 0.8 20 85.3 + 7.8 ( 19] a 7 9 . 4 + 4.9 [ 19] a 77.4 + 3.8 [ 18] a 7 4 . 5 + 7.4 t 19] a 70.6 + 5.2 l 18] a 68.6 + 7.6 76.7 + 5.3 DAYS = DAYS OF STUDY ( ) = NUMBER OF VALUES AVERAGED a. Excludes values that were associated with spillage. b. Last value recorded before cohabitation. * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 10 11 1 1.2 20 20 84.8 + 8.9 81.6 + 10.5 1 19] a 82.7 + 8.8* 77.4 + 6.9 75.1 + 7.8 ( 19] a 68.8 + 6.7 I 19] a 78.8 + 6.3 [ 19] a 85.8 + 6.6 7 8 . 6 + 4.9 77.9 + 3.4 [ 17] a 7 4 . 1 + 6.5 1 19] a 70.2 + 5.9 ( 19] a 6 5 . 8 + 6.7 [ 19] a 7 5 . 4 + 4.0 ( 19] a 12 13 1.6 2 28 28 88.4 + 7.7 78.9 + 5.6 77.3 + 5.3 [ 27] a 7 3 . 2 + 4.4 [ 27] a 70.8 + 6.8 65.2 + 4.4* 75.8 + 4.2 85.0 + 7.0 [ 27] a 76.6 + 4 .7* 75.0 + 5.2 ( 27] a 71.1 + 6.0* 68.2 + 4.9* ( 26] a 63.9 + 4.3** ( 27] a 73.4 + 3.8* t 271 a 418-018-.PAGE B000113 LvOo PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 11 (PAGE 2): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG /KG: PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID RATS TESTED N 28 29 INCLUDED IN ANALYSES N 28 28a FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 DAYS 8 - 15 MEAN+S.D. M E A N + S .D . 89.5 + 6.9 [ 26] b 85.6 + 6.6## 8 5 . 8 + 7.8 80.4 + 12.4* DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 42C DAYS 1 - 42c MEAN+S.D. MEAN+S. D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 82.4 + 7.0# [ 27]b 78.8 + 6.2# 74,6 + 6.5 [ 26] b 70.4 + 6.8 80.3 + 4.5# 76.9 + 7.9** [ 27] b 71.6 + 6.2** [ 27] b 7 1 . 9 + 6.3 6 7 . 3 + 5.6 [ 27] b 75.7 + 5.3** [ 27] b DAYS = DAYS OF STUDY [ = NUMBER OF VALUES AVERAGED a. Excludes values for rat 10936, which was found dead on day 2 of study, b. Excludes values that were not recorded or were associated with spillage. c. Last value recorded before cohabitation. # Significan! different from the Group 1 value (pcO.05) ## Significan! different from the Group 1 value (p<0.01) different from the Group 2 value (p<0.05) different from the Group 2 value (pcO.Ol) 4 2 + MEVALONIC ACID 28 28 8 6 . 8 + 5.6 [ 27] b 7 8 . 9 + 7.7** 7 6 . 9 + 7.5** f 27] b 71.8 + 5.6** ( 26] b 70.9 + 10.4 [ 26] b 65.6 + 6.5** 7 5 . 4 + 4.7** 418-018:PAGE B-40 000114 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 11 (PAGE 3): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS CHOLESTEROL CONTROL 1 . 6 CHOLESTEROL RATS TESTED N 28 28 FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 M E A N + S .D . 8 5 . 2 + 5.1 83.6 + 6.0 DAYS 8 - 15 DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 42c DAYS 1 - 42c MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 8 0 . 8 + 5.2 1 27) a 80.2 + 6.9 [ 27) b 76.2 + 6.3 [ 27] b 77.9 + 8.0## [ 25] a, b 7 1 . 9 + 6.5 ( 27) b 7 8 . 6 + 4.7 [ 27) b 79.0 + 4.0 76.7 + 6.9 71.4 + 3.9** 71.5 + 3.6** ( 23] a 67.2 + 4 .5** 75.1 + 2.8* DAYS = DAYS OF STUDY ( ) = NUMBER OF VALUES AVERAGED a. Excludes values that were associated with spillage. b. Excludes values for rat 11562, which was moribund sacrificed on day 16 of study. c. Last value recorded before cohabitation. ## Significantly different from the Group 1 value (p<0.01). * Significantly different from the Group 5 value (p<0.05). ** Significantly different from the Group 5 value (p<0.01). 2 + CHOLESTEROL 28 84.6 + 6.3 79.4 + 7.2 76.9 + 8.8 [ 27] a 70.3 + 6.6** 71.4 + 7.7** [ 25] a 67.6 + 6.5** 75.5 + 5.7* 418-018 :PAGE B~ 000115 PROTOCOL 418-018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 12 (PAGE 1) : MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 1 RATS TESTED N 28 20 20 20 PREGNANT N 27 20 17 19 MATERNAL FEED CONSUMPTION (G/DAY) +DAYS 0 - 7 MEAN+S.D. 22.3 2.2 22.7 + 2.3 20.6 + 2.4* 20.7 + 2.0* DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. 22.9 + 2.2 MEAN+S.D. 24.3 + 2.2 MEAN+S.D 23.2 + 1.9 23.2 + 1.6 [ 19] a 24.5 + 2.5 23.6 + 1.8 22.7 + 2.5 25.1 + 2.2 [ 13]a,b 22.8 + 2.2 t 13]a,b 21.7 + 2.7 25.0 + 2.5 22.5 + 1.8 DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that were not recorded as well as those associated with spillage. b. Excludes values for dams that delivered. * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 11 12 1.2 1.6 20 28 18 25 13 2 28 27 19.6 + 1.9** 22.0 + 2.1 19.0 + 21.2 + 2.1** 2.7 18.6 + 2.2** [ 26] a 2 1 . 0 + 4.0* 24.7 + 2.6 22.1 + 1.9 25.2 + 2.3 [ 23] b 21.8 + 2.1* ( 23] b 2 4 . 4 + 2.8 [ 26] a 21.4 + 2.0** [ 26] a siioeo 418-018 :PAGE B-42 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 12 (PAGE 2) : MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1 . 6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS TESTED N 28 28a 28 PREGNANT N 27 26 27 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 7 M E A N + S .D . 2 1 . 6 + 1.8 18.2 + 2.3** 19.1 + 2.7** DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. 2 3 . 2 + 2.1 [ 24]b,c 2 4 . 4 + 2.5 [ 26] b 23.0 + 1.8 [ 26] b 2 0 . 9 + 1.9** 25.0 + 2.4 [ 22] d 21.5 + 2.0** [ 22] d 20.8 + 2.0** [ 25] c 23.8 + 3.6 [ 24] d 21.5 + 2.2** t 24] d DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rat 10936, which was found dead prior to gestation. b. Excludes values for dam 10926, which was moribund sacrificed on day 12 of gestation accident). c. Excludes values that were associated with spillage. d. Excludes values for dams that delivered. ** Significantly different from the Group 2 value (p<0.01). (death was attributed to an intubation 418-018:PAGE B-43 000117 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 12 (PAGE 3): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL RATS TESTED N 27a 28 PREGNANT N 25 28 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 7 DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 2 2 . 9 + 2.9 ( 23] b 24.3 + 2.7 [ 24] b 2 5 . 4 + 2.9 ( 23] c 2 4 . 3 + 2.6 [ 23] c 1 8 . 9 + 2.2** 21.2 + 2.1** [ 27] b 24.6 + 2.1 [ 26] c 2 1 . 5 + 1.8** [ 26] c DAYS = DAYS OF GESTATION ( ] = NUMBER OF VALUES AVERAGED a. Excludes rat 11562, which was moribund sacrificed prior to gestation. b. Excludes values that were associated with spillage. c. Excludes values for dams that delivered. ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 28 27 18.3 + 2.0** 20.8 + 1.8** ( 26] b 25.0 + 2.3 [ 26] c 21.5 + 1.8** [ 26] c 418-018-.PAGE B-44 000118 1'ROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 13 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS i VEHICLE CONTROL 8 0.4 9 0.8 10 1 RATS TESTED N 28 20 20 20 PREGNANT N 27 20 17 19 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 MEAN+S.D. 77.2 + 5.7 7 6 . 3 + 5.0 72.2 + 5.4* 72.3 + 7.2* DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. 73.4 + 4.2 MEAN+S.D. 66.8 + 5.4 MEAN+S.D, 71.9 + 4.0 7 2 . 2 + 4.1 [ 19] a 6 5 . 1 + 6.8 [ 18] b 7 0 . 8 + 4.0 [ 18] b 74.0 + 4.2 68.8 + 4 .3 [ 11]a,b 70.5 + 3.9 ( 11]a,b 70.6 + 8.3 6 8 . 1 + 9.0 t 12] b 7 1 . 2 + 6.9 [ 12] b DAYS = DAYS OF GESTATION 1 ] = NUMBER OF VALUES AVERAGED a. Excludes values that were not recorded as well as those associated with spillage. b. Excludes values for dams that delivered. * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 11 12 1.2 1.6 20 28 18 25 13 2 28 27 70.3+ 73.7+ 5.2** 6.8 68.6+ 71.8+ 5.4** 6.3 69.0 + 6.6** 26] a 72.7 + 11.4 6 8 . 9 + 5.8 [ 15] b 7 0 . 4 + 4.5 [ 15] b 7 2 . 1 + 5.8* [ 20] b 7 0 . 8 + 3.6 [ 20] b 7 0 . 7 + 7.5 [ 20] a ,b 7 1 . 0 + 5.6 [ 20] a ,b PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 13 (PAGE 2): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS TESTED N 28 28a 28 PREGNANT N 27 26 27 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 M E A N + S .D . 75.4 + 6.1 6 6 . 3 + 6.4** 6 8 . 4 + 8.2** DAYS 7 - 14 DAYS 1 4 - 2 1 DAYS 0 - 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. 74.9 + 6.0 [ 24]b,c 67.9 + 4.8 ( 25] b, c 72.5 + 4.0 [ 25]b,c 71.5 + 3.4* 71.8 + 5.9 l 21] d 70.2 + 4.2 [ 21] d 70.1 + 6.4** [ 25] c 6 8 . 6 + 8.4 ( 19] d 70.0 + 5.9 [ 19] d DAYS = DAYS OF GESTATION ( ] = NUMBER OF VALUES AVERAGED a. Excludes rat 10936, which was found dead prior to gestation. b. Excludes values for rat 10926, which was moribund sacrificed on day 12 of gestation accident). c. Excludes values that were associated with spillage. d. Excludes values for dams that delivered. * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). (death was attributed to an intubation 418-018:PAGE B-46 000120 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 13 (PAGE 3) : MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL RATS TESTED N 27a 28 PREGNANT N 25 28 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 7 6 . 7 + 7.8 l 23) b 7 4 . 8 + 6.7 [ 24 ]b 66.9 + 6.4 [ 23] c 72.2 + 5.7 [ 23] c 6 9 . 1 + 6.0** 7 1 . 8 + 5.1 ( 27] b 71.2 + 4.4* [ 23 ]c 70.6 + 3.5 [ 23] c DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rat 11562, which was moribund sacrificed prior to gestation. b. Excludes values that were associated with spillage. c. Excludes values for dams that delivered. * Significantly different from the Group 5 value (p<0.0S). ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 28 27 67.5 + 7.2** 71.8 + 5.3 [ 26] b 7 2 . 6 + 6.0** [ 19] c 70.4 + 5.3 [ 19] c 418-018:PAGE B-47 000121 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 14 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS I VEHICLE CONTROL RATS TESTED N 19 PREGNANT N 19 DELIVERED A LITTER N 19 INCLUDED IN ANALYSES N 19 FEED CONSUMPTION (G/DAY) 8 0.4 20 20 20 20 9 0.8 20 17 17 17 10 ii 1 1.2 20 20 19 18 19 18 18a 18 12 1.6 20 17 17 14a 13 2 20 19 19 63 DAYS 1 - 5 MEAN+S.D. 28.8 + 4.7 ( 18] b 29.3 + 9.7 24.0 + 6.1 25.5 + 8.7 23.7 + 5.2 19.4 + 5.8** 17.2 7.3** DAYS = DAYS OF LACTATION ( ] = NUMBER OF VALUES AVERAGED a. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. b. Excludes values that were associated with spillage. ** Significantly different from the Group 1 value (p<0.01). 418-018:PAGE B-48 000122 f PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 14 (PAGE 2) : ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS TESTED N 19a 20a 20 PREGNANT N 18 18 19 DELIVERED A LITTER N 18 18 19 INCLUDED IN ANALYSES N 18 12b 3b FEED CONSUMPTION (G/DAY) DAYS I - 5 M E A N + S .D . 2 8 . 4 + 5.3 20.2 + 5.6** [ 11] c 1 3 . 9 + 2.2** DAYS = DAYS OF LACTATION ( ] = NUMBER OF VALUES AVERAGED a. Excludes rats that died or were moribund sacrificed prior to lactation. b. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. c. Excludes values that were associated with spillage. ** Significantly different from the Group 2 value (p<0.01). 418-018 (PAGE B-49 000123 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 14 (PAGE 3): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL RATS TESTED N 19a 20 20 PREGNANT N 17 20 19 DELIVERED A LITTER N n 20 19 INCLUDED IN ANALYSES N 17 13b 7b FEED CONSUMPTION (G/DAY) DAYS 1 - 5 MEAN+S.D. 31.6 + 5.1 19.9 + 6.1** 14.6 + 3.5** DAYS = DAYS OF LACTATION a. Excludes rat 11562, which was moribund sacrificed prior to lactation. b. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. ** Significantly different from the Group 5 value (p<0.01). 418-018:PAGE B-50 000124 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE IS (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 1 1.2 12 1.6 13 2 RATS TESTED N 19 20 20 20 20 20 20 PREGNANT N 19 20 17 19 18 17 19 DELIVERED A LITTER N 19 20 17 19 18 17 19 INCLUDED IN ANALYSES N 19 20 17 18a 18 14a 6a FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 5 M E A N + S .D . 92 .8 + 13.7 [ 18) b 92.3 + 32.1 77.5 + 15.0* 83.0 + 29.6* 78.5 + 15.3* 65.1 + 18.9** 60.7 + 26.6 DAYS = DAYS OF LACTATION ( ] = NUMBER OF VALUES AVERAGED a. Excludes values for dams that were sacrificed due to no surviving pups prior to day S of lactation. b. Excludes values that were associated with spillage. * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 418-018:PAGE B-5 000125 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 15 (PAGE 2) : RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID RATS TESTED N 19a 20a 20 PREGNANT N 18 18 19 DELIVERED A LITTER N 18 18 19 INCLUDED IN ANALYSES N 18 12b 3b FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 5 MEAN+S.D. 92.6 + 19.5 65.3 + 19.1** [ ll)c 4 6 . 9 + 6.2** DAYS = DAYS OF LACTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rats that died or were moribund sacrificed prior to lactation. b. Excludes values for dams that were sacrificed due to no surviving pups prior to day 5 of lactation. c. Excludes values that were associated with spillage. ** Significantly different from the Group 2 value (p<0.01). 9ZT000 418-018:PAGE B-52 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 15 (PAGE 3) : RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL RATS TESTED N 19a 20 20 PREGNANT N 17 20 19 DELIVERED A LITTER N 17 20 19 INCLUDED IN ANALYSES N 17 13b 7b FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 5 MEAN+S.D. 98.4 + 11.5 66.5 + 20.1** 52.0 + 14.7** DAYS = DAYS OF LACTATION a. Excludes rat 11562, which was moribund sacrificed prior to lactation. b. Excludes values for dams that were sacrificed due to no surviving pups ** Significantly different from the Group 5 value (p<0.01). prior to day 5 of lactation. 2T000 418-018:PAGE B-53 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 16 (PAGE 1): MATING AND FERTILITY - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 18 VEHICLE CONTROL. 0.4 9 0.8 10 11 12 1 1.2 1,6 13 2 RATS IN COHABITATION N 28 20 20 20 20 28 28 DAYS IN COHABITATION a M E A N + S .D 2.6 + 2.4 2.7 + 1.0 3 . 2 + 2.9 3 . 0 + 3.0 2 . 5 + 2.9 3 . 1 + 3.2 2 . 6 + 2.1 RATS THAT MATED N (% ) 28(100.0) 20(100.0) 19 ( 95.0) 20(100.0) 19( 95.0) 27( 96.4) 28(100.0) FERTILITY INDEX b N/N (%) 27/ 28 ( 96.4) 20/ 20 (100.0) 17/ 19 ( 89.5) 19/ 20 ( 95.0) 18/ 19 ( 94.7) 25/ 27 ( 92.6) 27/ 28 ( 96.4) RATS WITH CONFIRMED MATING DATES N 28 20 19 20 19 27 28 MATED BY FIRST MALE c DAYS 1-7 N (% ) 27( 96.4) 20(100.0) 19(100.0) 19( 95.0) 19(100.0) 26( 96.3) 27( 96.4) MATED BY SECOND MALE c DAYS 8-14 N (% M 3.6) 0 ( 00.0) 0( 00.0) M 5.0) 0( 00.0) M 3.7) M 3.6) RATS PREGNANT/RATS IN COHABITATION N/N 27/ 28 20/ 20 17/ 20 19/ 20 18/ 20 25/ 28 27/ 28 <%) ( 96.4) (100.0) ( 85.0) ( 95.0) ( 90.0) ( 89.3) ( 96.4) a. Restricted to rata with a confirmed mating date and rats that did not mate. b. Number of pregnancies/number of rats that mated. c. Restricted to rats with a confirmed mating date. 418-018:PAGE B-54 000128 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 16 (PAGE 2): MATING AND FERTILITY - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID RATS IN COHABITATION N 28 2 8a DAYS IN COHABITATION b M E A N + S .D . 2 . 8 + 2.2 2 . 0 + 1.1 RATS THAT MATED N (% ) 28(100.0) 28(100.0) FERTILITY INDEX c N/N (%) 27/ 28 ( 96.4) 26/ 28 ( 92.8) RATS WITH CONFIRMED MATING DATES N 28 28 MATED BY FIRST MALE d DAYS 1-7 N (% ) 27( 96.4) 28(100.0) MATED BY SECOND MALE d DAYS 8-14 N(%) 1( 3.6) 0( 00.0) RATS PREGNANT/RATS IN COHABITATION N/N <%> 27/ 28 ( 96.4) 26/ 28 ( 92.8) a. Excludes rat 10936, which was found dead on day 2 of study. b. Restricted to rats with a confirmed mating date and rats that did not mate. c. Number of pregnancies/number of rats that mated. d. Restricted to rats with a confirmed mating date. 4 2 + MEVALONIC ACID 28 2 . 8 + 2.2 28(100.0) 27/ 28 ( 96.4) 28 27( 96.4) M 3.6) 27/ 28 ( 96.4) 418-018:PAGE B-55 000129 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 16 (PAGE 3): MATING AND FERTILITY - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL RATS IN COHABITATION N 27a 28 DAYS IN COHABITATION b M E A N + S .D . 3 . 2 + 2.3 2 . 0 + 1.2** RATS THAT MATED N (% ) 27(100.0) 28(100.0) FERTILITY INDEX c N/N (%) 25/ 27 ( 92.6) 28/ 28 (100.0) RATS WITH CONFIRMED MATING DATES N 27 28 MATED BY FIRST MALE d DAYS 1-7 N (%) 26( 96.3) 28(100.0) MATED BY SECOND MALE d DAYS 8-14 Nil) 1! 3.7) 0( 00.0) RATS PREGNANT/RATS IN COHABITATION N/N (%) 25/ 27 ( 92.6) 28/ 28 (100.0) a. Excludes rat 11562, which was moribund sacrificed on day 16 of study. b. Restricted to rats with a confirmed mating date and rats that did not mate. c. Number of pregnancies/number of rats that mated. d. Restricted to rats with a confirmed mating date. ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 28 2 . 7 + 2.3 28 (100.0) 27/ 28 ( 96.4) 28 27{ 96.4) M 3.6) 27/ 28 ( 96.4) 418-018:PAGE B-56 000130 PROTOCOL 418*018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 17 (PAGE 1) : CAESAREAN-SECTIONING OBSERVATIONS * SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED N PREGNANT N (% ) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N CORPORA LUTEA M E A N + S .D . IMPLANTATIONS M E A N + S .D . LITTER SIZES M E A N + S .D . 1 VEHICLE CONTROL 9 8( 88.9) 8 16.1 + 14.9+ 13.5+ 1.6 1.2 1.3 12 1.6 B 8(100.0) 8 15.4 + 2.7 1 4 . 0 + 2.9 13.0 + 2.8 13 2 8 8(100.0) 8 18.9+ 15.2+ 14.9+ 5.1 1.8 1.7 418-018:PAGE B-57 000131 LIVE FETUSES N M E A N + S .D . 108 13.5 + 1.3 DEAD FETUSES N MEAN+S.D. 0 0.0 + 0.0 RESORPTIONS M E A N + S .D . 1.4 + 1.1 EARLY RESORPTIONS N M E A N + S .D . 11 1 .4 + 1.1 LATE RESORPTIONS N M E A N + S .D . 0 0.0+ 0.0 DAMS WITH ANY RESORPTIONS N(%) 8(100.0) DAMS WITH ALL CONCEPTUSES DEAD OR RESORBED N (% ) 0( 0.0) DAMS WITH VIABLE FETUSES N(%) 8(100.0) PLACENTAE APPEARED NORMAL N (% ) 8(100.0) * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 103 12.9+ 2.7 1 0.1+ 0.4 1.0 + 0.9 7 0.9+ 1.0 1 0.1 + 0.4 6( 75.0) 0( 0.0) 8(100.0) 8(100.0) 119 14.9+ 1.7 0 0.0 + 0.0 0 . 4 + 0.5* 3 0.4+ 0.5* 0 0.0+ 0.0 3( 37.5)** 0( 0.0) 8(100.0) 8(100.0) PROTOCOL 418018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 17 (PAGE 2): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED N PREGNANT N (%) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N CORPORA LUTEA MEAN+S.D. IMPLANTATIONS M E A N + S .D . LITTER SIZES M E A N + S .D . 2 MEVALONIC ACID CONTROL B 8(100.0) 3 1.6 + MEVALONIC ACID 8 8(100.0) 8 19.1 + 3.3 15.4 + 1.1 1 3 . 6 + 2.7 8 17.5 + 4 .1 15.5 + 2.4 13.5 + 3.1 4 2 + MEVALONIC ACID a 8(100.0) 8 18.2 + 2.6 16.1 + 1.0 1 3 . 6 + 2.4 LIVE FETUSES N MEAN+S.D. DEAD FETUSES N RESORPTIONS M E A N + S .D . EARLY RESORPTIONS N M E A N + S .D . LATE RESORPTIONS N M E A N + S .D . DAMS WITH ANY RESORPTIONS N (%) DAMS WITH ALL CONCEPTUSES DEAD OR RESORBED N (%) DAMS WITH VIABLE FETUSES N (% ) PLACENTAE APPEARED NORMAL N (% ) 109 13.6 + 2.7 0 1.8 + 2.2 14 1.8 + 2.2 0 0.0 + 0.0 5( 62.5) 0( 0.0) 8(100.0) 8(100.0) 108 13.5 + 3.1 0 2.0 + 2.6 15 1.9 + 2.6 i 0.1 + 0.4 6( 75.0) 0( 0.0) 8(100.0) 8(100.0) 109 13.6 + 2.4 0 2.5 + 3.0 19 2.4+ 3.0 1 0.1 + 0.4 6( 75.0) 0( 0.0) 8(100.0) 8 (100.0) 418-018:PAGE B-58 000132 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-S295.25) TABLE 17 (PAGE 3): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED N PREGNANT N (% ) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N CORPORA LUTEA M E A N + S .D . IMPLANTATIONS MEAN+S.D. LITTER SIZES M E A N + S .D . 5 CHOLESTEROL CONTROL 8 8(100.0) 8 1 7 . 8 + 2.7 15.9 + 2.2 15.2 + 2.3 6 1.6 + CHOLESTEROL 8 8(100.0) 8 17.6 + 3.7 1 4 . 6 + 4.2 1 4 , 0 + 4.2 7 2 + CHOLESTEROL 8 8(100.0) 6 1 8 . 1 + 6.8 1 3 . 2 + 4.4 1 1 . 5 + 3.9 LIVE FETUSES N M E A N + S .D . DEAD FETUSES N RESORPTIONS MEAN+S.D. EARLY RESORPTIONS N M E A N + S .D . LATE RESORPTIONS N DAMS WITH ANY RESORPTIONS N (% ) DAMS WITH ALL CONCEPTUSES DEAD OR RESORBED N{%) DAMS WITH VIABLE FETUSES N (% ) PLACENTAE APPEARED NORMAL N (% ) 122 15.2 + 2.3 0 0.6 + 0.5 5 0.6+ 0.5 0 5( 62.5) 0( 0.0) 8(100.0) 8(100.0) 112 14.0+ 4.2 0 0 . 6 + 1.1 5 0.6 + 1.1 0 3( 37.5) 0( 0.0) 8(100.0) 8(100.0) 92 11.5+ 0 1.8+ 14 1.8 + 0 3.9 1.4 1.4 6( 75.0) 0( 0.0) 8(100.0) 8(100.0) 418-018:PAGE B-59 000133 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 18 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL LITTERS WITH ONE OR MORE LIVE FETUSES N 8 IMPLANTATIONS M E A N + S .D . 14.9 + 1.2 LIVE FETUSES N M E A N + S .D . 108 13.5 + 1.3 POOLED FETAL BODY WEIGHTS (GRAMS)/LITTER MEAN+S.D. 70.3 + 4.7 % DEAD OR RESORBED CONCEPTUSES/LITTER M E A N + S .D . 9.1 + 6.4 * Significantly different from the Group 1 value (p<0.05). 12 1.6 8 14.0 + 2.9 103 12.9 + 2.7 65.9 + 15.5 8 . 0 + 5.0 13 2 8 15.2 + 119 14.9 + 1.8 1.7 74.8 + 8.8 2.4 + 3.4* 418-018 :PAGE B-60 000134 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 18 (PAGE 2): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY - Fo- GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS LITTERS WITH ONE OR MORE LIVE FETUSES IMPLANTATIONS N MEAN+S.D. 2 MEVALONIC ACID CONTROL 8 15.4 + 1.1 3 1.6 + MEVALONIC ACID 8 15.5 + 2.4 4 2 + MEVALONIC ACID 8 16.1 + 1.0 LIVE FETUSES N MEAN+S.D-. POOLED FETAL BODY WEIGHTS (GRAMS)/LITTER M E A N + S .D . % DEAD OR RESORBED CONCEPTUSES/LITTER MEAN+S.D. 109 13.6 + 2.7 70.9 + 15.0 11.6 + 14.9 108 13.5 + 3.1 66.4 + 14.7 12.5 + 15.0 109 13.6 + 2.4 69.3 + 16.7 14.9 + 17.3 '-9HDVd:8lO-81fr 000135 0\ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 18 (PAGE 3) : LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS LITTERS WITH ONE OR MORE LIVE FETUSES IMPLANTATIONS N M E A N + S .D . 5 CHOLESTEROL CONTROL 8 15.9 + 2.2 6 1.6 + CHOLESTEROL 8 14.6 + 4 .2 7 2 + CHOLESTEROL 8 13.2 + 4.4 LIVE FETUSES N M E A N + S .D . 122 15.2 + 2.3 POOLED FETAL BODY WEIGHTS (GRAMS)/LITTER MEAN+S.D. 79.4 + 11.8 % DEAD OR RESORBED CONCEPTUSES/LITTER M E A N + S .D . 4.0 + 3.4 * Significantly different from the Group 5 value (p<0. 05) . 112 14.0 + 4.2 70.6 + 22.9 4.1 + 7.4 92 11.5 + 3 .9 56.2 + 18.3 12.0 + 10.0* 418-018:PAGE B-62 000136 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 19 (PAGE 1): NATURAL DELIVERY OBSERVATIONS - SUMMARY Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1B VEHICLE CONTROL. 0.4 RATS ASSIGNED TO NATURAL DELIVERY N 19 20 PREGNANT N (% ) 19(100.0) 20(100.0) DELIVERED LITTERS N (% ) 19(100.0) 20(100.0) DURATION OF GESTATION a M E A N + S .D . 22.9 + 0.3 22.6 + 0.5 9 0.8 10 11 12 1 1.2 1.6 13 2 20 20 20 20 20 17( 85.0) 19( 95.0) 18( 90.0) 17( 85.0) 19( 95.0) 17(100.0) 19(100.0) 18(100.0) 17(100.0) 19(100.0) 22.5 + 0.5* 22.4 + 0.6** 22.3 + 0.5** 22.0 + 0.0** 22.2 + 0.4** IMPLANTATION SITES PER DELIVERED LITTER N MEAN+S.D. 279 232 257 303 275 243 274 1 4 . 7 + 2.3 16.2 + 1.8 1 5 . 1 + 2.2 1 5 . 9 + 2.0 15.3 + 2.5 1 4 . 3 + 2.1 1 4 . 4 + 1.9 DAMS WITH STILLBORN PUPS N (% ) 4( 21.0) 8 ( 40.0)** 2 ( 11.8) M 5.3)* K 5.6)* K 5.9)* 1( 5.3)* DAMS WITH NO LIVEBORN PUPS N (%) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) GESTATION INDEX b % N/N 100.0 19/ 19 100.0 20/ 20 100.0 17/ 17 100.0 19/ 19 100.0 18/ 18 100.0 17/ 17 100.0 19/ 19 DAMS WITH ALL PUPS DYING DAYS 1-5 POSTPARTUM N (%) 0( 0.0) 0( 0.0) 0( 0.0) 1( 5.3) 0( 0.0) 4 ( 23.5) 14 ( 73.7)** a. Calculated as the time (in days) elapsed between confirmed mating (arbitrarily defined as 0 hour) and the time (in days) the first pup was delivered. b. Number of rats with live offspring/number of pregnant rats. * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 418-018:PAGE B-63 000137 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 19 (PAGE 2): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS ASSIGNED TO NATURAL DELIVERY N PREGNANT N (% ) DELIVERED LITTERS N (% ) DURATION OF GESTATION b M E A N + S .D . 2 MEVALONIC ACID CONTROL 19a 18 ( 94.7) 18(100.0) 22.7 + 0.5 3 1.6 + MEVALONIC ACID 20a 18( 90.0) 18(100.0) 2 2 . 2 + 0.4* 4 2 + MEVALONIC ACID 20 19( 95.0) 19(100.0) 2 2 . 0 + 0.4** IMPLANTATION SITES PER DELIVERED LITTER N M E A N + S .D . 263 14.6 + 2.0 251 1 3 . 9 + 2.2 289 1 5 . 2 + 1.4 DAMS WITH STILLBORN PUPS N(%) 2 ( 11.1) 4 ( 22.2) 2( 10.5) DAMS WITH NO LIVEBORN PUPS N (%) 0( 0.0) 0( 0.0) 0( 0.0) GESTATION INDEX C % N/N 100.0 18/ 18 100.0 18/ 18 100.0 19/ 19 DAMS WITH ALL PUPS DYING DAYS 1-5 POSTPARTUM N (% ) 0( 0.0) 6( 33.3) 17 ( 89.5)** a. Excludes values lor rats that died or were moribund sacrificed prior to lactation. b. Calculated as the time (in days) elapsed between confirmed mating (arbitrarily defined as 0 hour) and the time (in days) the first pup was delivered. c. Number of rats with live offspring/number of pregnant rats. * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). 418-018(PAGE B-64 000138 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 19 (PAGE 3): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS ASSIGNED TO NATURAL DELIVERY N PREGNANT N (% ) DELIVERED LITTERS N (% ) DURATION OF GESTATION b M E A N + S .D . 5 CHOLESTEROL CONTROL 19a 17 ( 89.5) 17(100.0) 22.8 + 0.4 6 1.6 + CHOLESTEROL 20 20(100.0) 20(100.0) 22.0 + 0.4** 7 2 + CHOLESTEROL 20 19( 95.0) 19(100.0) 22.3 + 0.4** IMPLANTATION SITES PER DELIVERED LITTER N M E A N + S .D . 241c 1 5 . 1 + 1.4 ( 16] c 288 1 4 . 4 + 1.7 291 1 5 . 3 + 2.0 DAMS WITH STILLBORN PUPS N (% ) 2 ( 11.8) 2( 10.0) 2( 10.5) DAMS WITH NO LIVEBORN PUPS N(%) 0( 0.0) 0( 0.0) M 5.3) GESTATION INDEX d % N/N 100.0 17/ 17 100.0 20/ 20 94.7 18/ 19 DAMS WITH ALL PUPS DYING DAYS 1-5 POSTPARTUM ' N (% ) 0( 0.0) 8 ( 40.0) 14 ( 73.7)** a. Excludes rat 11562, which was moribund sacrificed prior to lactation. b. Calculated as the time (in days) elapsed between confirmed mating (arbitrarily defined as 0 hour) and the time (in days) the first pup was delivered. c. Excludes a value for dam 10969, which appeared incorrectly recorded. d. Number of rats with live offspring/number of pregnant rats. ** Significantly different from the Group 5 value (p<0.01). 418-018:PAGE B-65 000139 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 1): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 1 1.2 12 1.6 13 2 DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 19 20 17 19 18 17 19 PUPS DELIVERED (TOTAL) N M E A N + S .D . 265 300 247 287 253 231 252 13.9 + 2.6 15.0 2.3 14.5 + 2.3 15.1 + 2.3 14.0 + 2.9 13.6 + 2.8 13.3 + 2.5 LIVEBORN STILLBORN UNKNOWN VITAL STATUS MEAN+S.D. 13.7 2,7 14.6 + 2.4 14.4 + 2.4 15.0 + 2.3 14.0 + 2.9 13.4 + 2.9 13.2 + 2 .5 N (% ) 260 ( 98. 1) 291 ( 97.,0) 245 ( 99. 2) 285 ( 99. 3) 252 ( 99. 6) 227 ( 98 .3) 251 ( 99. 6) MEAN+S.D. N(%) 0.3 + 0.6 5 ( 1.9) 0.4 + 0.6 9 1 3.0) 0.1 + 0.3 2( 0.8) 0.1 + 0.4 2( 0.7) 0.0 + 0.2 K 0.4) 0.2 + 1.0 4( 1.7) 0.0 + 0.2 K 0.4) N000000 0 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2- 5 VIABILITY INDEX a N / N (%) N / N (%) % N/N 1/260( 0.4) 3/291( 1.0) 1/245 ( 0.4) 4/285 ( 1.4) 6/252( 2.4) 13/227 ( 5.7) 56/251(22.3)** 6/259( 2.3) 4/288( 1.4) 16/244( 6.6) 28/281(10.0) 40/246(16.3) 102/214(47.7)** 152/195(77.9)** 97.3 253/260 97.6 284/291 93.1 228/245 CD CD CD 253/285 81.7 206/252 49.3 112/227** 17.1 43/251** D A Y (S ) = D A Y (S ) POSTPARTUM a. Number of live pups on day 5 postpartum/number of liveborn pups on day 1 postpartum. ** Significantly different from the Group 1 value (p<0.01). 418-018:PAGE B-66 000140 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 20 (PAGE 2): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 1 1.2 12 1.6 13 2 DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 19 20 17 19 18 17 19 SURVIVING PUPS/LITTER a DAY lb M E A N + S .D . 13.7 + 2.7 14 .6 + 2.4 14.4 + 2.4 15.0 + 2.3 14.0 + 2.9 13.4 + 2.9 13.2 + 2.5 DAY 2 DAY 3 M E A N + S .D . 13 .5 + 2.5 14 .4 + 2.4 13.9 + 2.2 1 3 . 8 + 3.0 12.9 + 3 .2 MEAN+S.D. 13.4 + 2.5 14.3 + 2.4 13.6 + 1.8 1 3 . 5 + 4.0 12.1 + 3.3 8.0 + 5.7* 3 .4 + 4.4** 7.1 + 5.5** 2.5 + 4.5** DAY 4 MEAN+S.D. 13.4 + 2.4 14.2 + 2.4 13.5 + 2.0 13.5 + 4.0 11.6 + 3.4 6.8 + 5.5** 2.4 + 4.4** DAY 5 M E A N + S .D . 13.3 + 2.4 14.2 + 2.4 13.4 + 2.1 1 3 . 3 + 4.0 11.4 + 3 .5 6.6 + 5.6** 2.3 + 4.4** 418-018:PAGE B-67 000141 PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY lb M E A N + S .D . 50.0 + 17.4 46.4 + 14.1 53.9 + 11.3 46.9 + 15.9 50.0 + 14.1 45.8 + 9.6 50.1 + 14.0 DAY 2 DAY 3 DAY 4 DAY 5 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 50.3 + 17.1 50.4 + 17.0 50.6 + 17.0 50.7 + 16.9 46.0 + 13.8 46.2 + 13 .7 46.1 + 13 .8 46.1 + 13.8 54.0 + 11.6 54 .5 + 11.0 54.7 + 11.1 54.9 + 10.7 48.5 + 17.0 49.6 + 14 .6 46.6 + 15.2 l 18] c 46.3 + 15.1 [ 18] c 46.5 + IS.3 [ 18] c 48.5 + 15.0 48.5 + 14.6 47.7 + 15.9 46.0 + 21.3 t 14]1c 46.1 + 27.4 l 14] C 46.1 + 26.7 [ 14] c 39.1 + 25.1 l 13] c 59.4 + 25.1 [ 11] c 51.1 + 33.1 [ 7] C 43.5 + 27.3 t 6} c 42.2 + 30.3 t 5] C DAY = DAY POSTPARTUM [ ] = NUMBER OF VALUES AVERAGED a. Average number of live pups per litter, including litters with no surviving pups. b. Includes pups born alive, found dead day 1 postpartum. c. Excludes values for litters with no surviving pups. * Significantly different from the Group 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 3) : LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0..8 10 11 1 1.2 12 1.6 13 2 DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 19 20 17 19 IB 17 19 POOLED PUP WEIGHTS/LITTER (GRAMS) DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAYS 1 - 5 (BODY WEIGHT CHANGE) M E A N + S .D . MEAN+S.D. MEAN+S.D. M E A N + S .D . MEAN+S.D. MEAN+S.D. 86.9 + 15.4 85.2 + 15.1 84.2 + 10.4 85.8 + 14.3 77.2 + 16.2 67.0 + 15.9** 58.5 + 25.6** [ 18] a 94.4 + 15.1 92.0 + 14.7 87.8 + 12.1 84.2 + 20.2 74.5 + 20.2** 52.8 + 26.0** 33.5 + 28.7** ( 14] a (H U 103.8 + 16.1 99.5 + 17.4 94.0 + 13.8 95.1 + 15.7 76.0 + 25.4** 51.5 + 30.5** 44.4 + 36.3** [ 18] a [ 14] a i 7] a 116.6 + 18.2 109.5 + 19.7 104.2 + 17.9 105.1 + 17.6 79.2 + 28.2** 55.0 + 34.8** 54.1 + 38.7** [ 18] b [ 18] a [ 14] a [ 6]a 129.4 + 19.8 121.5 + 21.2 112.6 + 20.8 113.0 + 19.1 85.2 + 32.2** 63.5 + 37.4** 68.3 + 38.8** l 18] a ( 13] a [ 5] a +42.5 + 12.4 +36.3 + 12.7 +28.5 + 17.5 +25.9 + 11.8* *+7.9 + 26.5** -3.2 + 33.0** -9.1 + 35.7* [ 18] a [ 13] a [ 5] a AVERAGE PUP WEIGHT (GRAMS) DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN+S.D. 6.4 + 0.4 M E A N + S .D . 7.1 + 0.6 M E A N + S .D . 7.8 + 0.7 M E A N + S .D . 8,8 + 0.9 M E A N + S .D . 9.8 + 1.1 M E A N + S .D . +3.4 + 0.9 5.9 + 0.6* 5.9 + 0.6* 5.8 + 0.8** 5.7 + 0.6** 5 . 4 + 0 . 5 * * 6.4 + 0.5* 6.4 7.0 + 0.7* * 6.9 7.8 + 0.9* 7.8 E 19] b 8.6 + 1.0* * 8.5 +2.7 + 0.7* +2.5 + 0.8** + 1.0** + 1.2* + 1.4** + 0.9* 6.1 + 0.8** 6.7 + 0.9** [ 18] a 7.5 + 1.1** [ 18] a 8.1 + 1.3** [ 18] a +2.2 + 0.8** ( 18] a 5.8 6.2 6.8 7.4 + 1.7 + 0.7** 5.4 + 0.5** [ 14] a + 1.0** 5.8 + 0.8** ( 14] a + 1.1** 6 . 3 + 1 . 0 * * [ 14] a + 1.4** 7.1 + 1.1** t 13] a + 1.2** 1 +1.7 + 1.0** E 13 ]a 5.3 + 0.5** [ 18] a 5.2 + 0.8** [ Hla 5.9 + 1.1** [ 7] a 6.5 + 1.4** [ 6]a 7.4 + 1.5** [ 5] a +1.5 + 1.2** [ 5]a DAY = DAY POSTPARTUM [ ] = NUMBER OF VALUES AVERAGED a. Excludes values for litters with no surviving pups. b. Excludes values that were not recorded. * Significantly different from the Group 1 value (p<0,05). * * Significantly different from the Group 1 value (p<0.01). 000142 89-9 3 0 Vd: 8 10-81^ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 4): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1 . 6 + MEVALONIC ACID DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 18 18 PUPS DELIVERED (TOTAL) N M E A N + S .D . 237 1 3 . 2 + 2.2 233 1 2 . 9 + 2.3 LIVEBORN MEAN+S.D. N (% ) 1 3 . 0 + 2.2 234( 98.7) 1 2 . 6 + 2.1 227( 97.4) STILLBORN MEAN+S.D. N (%) 0.2 + 0.5 3 ( 1.3) 0.3 + 0.7 6( 2.6) UNKNOWN VITAL STATUS N 0 0 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2- 5 N / N {%) N / N (%) 1/234 ( 0.4) 2/233( 0.8) 22/227( 9.7) 111/205( 54.1) VIABILITY INDEX a % N/N 98.7 231/234 41.4 94/227* D A Y (S ) = DAY(S) POSTPARTUM a. Number of live pups on day 5 postpartum/number of liveborn pups on day 1 postpartum, * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). 4 2 + MEVALONIC ACID 19 270 14.2 + 1.4 1 4 . 1 + 1.4 268( 99.2) 0 . 1 + 0.3 2( 0.7) 0 93/268( 34.7)** 172/17 5 ( 98.3)** 1.1 3/268** 418-018:PAGEB-69 000143 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 5): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N SURVIVING PUPS/LITTER a DAY lb M E A N + S .D . DAY 2 M E A N + S .D . DAY 3 M E A N + S .D . DAY 4 M E A N + S .D . DAY 5 M E A N + S .D . 2 MEVALONIC ACID CONTROL 18 3 1.6 + MEVALONIC ACID 18 13.0 + 2.2 12.9 + 2.2 12.8 + 2.3 12.8 + 2.3 12.8 + 2.3 12.6 + 2.1 6.2 + 4.6** 5 . 6 + 4.8** 5.2 + 4.8** 5 . 2 + 4.8** 4 2 + MEVALONIC ACID 19 14 .1 + 1.4 1.4 + 2.0** 0.5 + 1.1** 0.3 + 0.8** 0.2 + 0.5** PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY lb DAY 2 DAY 3 DAY 4 DAY 5 M E A N + S .D . MEAN+S .D . MEAN+S.D. MEAN+S.D. MEAN+S.D. 56.2 + 12.4 56.4 + 12.2 55.8 + 12.3 55.8 + 12.3 55.8 + 12.3 49.9 + 10.8 55.0 + 18.0 [ 15] C 49.8 + 12.4 ( 13 ]C 52.2 + 10.4 [ 12] c 52.2 + 10.4 [ 12 ]c DAY = DAY POSTPARTUM [ ) = NUMBER OF VALUES AVERAGED a. Average number of live pups per litter, including litters with no surviving pups. b. Includes pups born alive, found dead day 1 postpartum. c. Excludes values for litters with no surviving pups. ** Significantly different from the Group 2 value (p<0.01). 48.0 + 15.6 54.6 [ 23.3 [ 55.6 t 50.0 1 + 41.1 9] C + 32.5** 5] c + 50.9 3] c + 70.7 2]c 418-018:PAGE B-70 000144 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 6) : LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 18 POOLED PUP WEIGHTS/LITTER (GRAMS) DAY 1 MEAN+S.D. 83.0 + 11.5 DAY 2 M E A N + S .D . 90.6 + 11.6 DAY 3 MEAN+S.D. 99.7 + 14.4 DAY 4 M E A N + S .D . 112.1 + 15.5 DAY 5 MEAN+S.D. 123.9 + 17,1 DAYS 1 - 5 (BODY WEIGHT CHANGE) M E A N + S .D . +40.9 + 8.2 AVERAGE PUP WEIGHT (GRAMS) DAY 1 M E A N + S .D . 6.4 + 0.5 DAY 2 MEAN+S.D. 7.1 + 0.6 DAY 3 MEAN+S.D. 7.8 + 0.7 DAY 4 MEAN+S.D. 8.8 + 0.8 DAY 5 M E A N + S .D . 9.8 + 1.0 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN+S .D . +3.3 + 0.8 DAY = DAY POSTPARTUM [ ] = NUMBER OF VALUES AVERAGED a. Excludes values for litters with no surviving pups. b. Excludes a value that was not recorded. * Significantly different from the Group 2 value (p<0.05). ** Significantly different from the Group 2 value (p<0.01). 3 1.6 + MEVALONIC ACID 18 63.1 + 18.7** 43.6 + 25.9** [ 15] a 50.1 + 28.6** [ 13) a 60.6 + 35.9** [ 12] a 6 3 . 8 + 33.4** [ 12] a -5.6 + 32.5** ( 12] a 5.6 + 0.5** 5.6 + 0.8** [ 15] a 6.1 + 1.2** [ 13]a 7.2 + 1.7** ( 12] a 7.7 + 1.8** ( 12] a + 2 . 0 + 1.4** l 12] a 4 2 + MEVALONIC ACID 19 57.1 + 22.2** t 16] a 17.5 + 9.8** [ 8] a, b 11.0 + 9.1** [ 5] a 12.1 + 7.1** [ 3]a 11.0 + 8.9** [ 2 ]a -39.7 + 33.5 [ 2]a 5.2 + 0.3** [ 16] a 6.8 + 5.4** t 8] a,b 5.1 + 1.1** [ 5] a 5.8 + 2.2** [ 3 3a 6.7 + 2.8** t 2] a +1.3 + 2.6* [ 2] a 418-018:PAGE B-71 000145 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 7): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL e 1.6 + CHOLESTEROL DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 17 20 PUPS DELIVERED (TOTAL) N M E A N + S .D . 247 14.5 + 2.2 271 1 3 . 6 + 2.1 LIVEBORN MEAN+S.D. N (% ) 1 4 . 4 + 2.2 24 5 ( 99.2) 13.4 + 2.3 269( 99.3) STILLBORN MEAN+S.D. N(%) 0.1 + 0.3 2( 0.8) 0.1 + 0.3 2 ( 0.7) UNKNOWN VITAL STATUS N 0 0 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2- 5 N/N{%) N / N (%) 0/245( 0.0) 5/245( 2.0) 18/2691 6.7)* 138/251( 55.0)* VIABILITY INDEX a % N/N 98.0 240/245 42.0 113/269* DAY(S) = D A Y (S ) POSTPARTUM a. Number of live pups on day 5 postpartum/number of liveborn pups on day 1 postpartum. * Significantly different from the Group 5 value (p<0.05). ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 18 261 1 4 . 5 + 1.7 1 4 . 4 + 1.8 259( 99.2) 0 . 1 + 0.3 2 ( 0.8) 1 27/258( 10.5)** 194/231( 84.0)** 14.3 37/258** 418-018:PAGE B-72 9*1000 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 8): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N SURVIVING PUPS/LITTER a DAY lb M E A N + S .D . DAY 2 MEAN+S.D. DAY 3 M E A N + S .D . DAY 4 M E A N + S .D . DAY 5 M E A N + S .D . 5 CHOLESTEROL CONTROL 17 14.4 + 2.2 14.3 + 2.2 14 .2 + 2.1 14.2 + 2.1 14.1 + 2.0 6 1.6 + CHOLESTEROL 20 13.4 + 2.3 8.4 + 4.6** 6.0 + 5.5** 5.8 + 5.4** 5.6 + 5.5** 7 2 + CHOLESTEROL 18 14.4 + 1.8 4.0 + 4.4** 2.2 + 4.0** 2.2 + 3.9** 2.0 + 4.0** PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY lb DAY 2 DAY 3 DAY 4 DAY 5 MEAN+S .D . MEAN+S.D. MEAN+S.D. MEAN+S.D. M E A N + S .D . 48.3 + 14 .5 48.2 + 14.9 48.2 + 14.8 48.2 + 14 .8 48.4 + 14 .9 47.3 + 16.8 50.0 + 24.0 [ 191 C 41.1 + 26.8 [ 14] c 44.4 + 25.0 t 13] C 43.9 + 26.0 [ 12] c DAY = DAY POSTPARTUM [ ] = NUMBER OF VALUES AVERAGED a. Average number of live pups per litter, including litters with no surviving pups. b. Includes pups born alive, found dead day 1 postpartum. c. Excludes values for litters with no surviving pups. ** Significantly different from the Group 5 value (p<0.01). 56.1 + 11.9 59.8 + 32.2 ( 14] C 49.6 + 33.9 [ 7]c 48.6 + 33.8 [ 7] c 48.0 + 21 .4 l 5] c 418-018:PAGE B-73 000147 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 20 (PAGE 9) : LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 17 POOLED PUP WEIGHTS/LITTER (GRAMS) DAY 1 M E A N + S .D . 90.9 + 12.5 DAY 2 MEAN+S.D. 100.7 + 18.9 DAY 3 MEAN+S.D. 108.7 + 1 6 . 8 DAY 4 MEAN+S.D. 123.4 + 17.7 DAY 5 DAYS: i - 5 (BODY WEIGHT CHANGE) MEAN+S.D. MEAN+S.D. 134.4 + 20.3 [ 16] b +44.4 + 11.8 [ 161b AVERAGE PUP WEIGHT (GRAMS) DAY 1 M E A N + S .D . 6.3 + 0.5 DAY 2 MEAN+S.D. 7.1 + 0.8 DAY 3 M E A N + S .D . 7.7 + 0.8 DAY 4 MEAN+S.D. 8.8 + 0.9 DAY 5 DAYS 1 - 5 (BODY WEIGHT CHANGE) M E A N + S .D . M E A N + S .D . 9.6 + 1.1 [ 16] b + 3 . 3 + 0.7 ( 16] b DAY = DAY POSTPARTUM [ ] = NUMBER OF VALUES AVERAGED a. Excludes values for litters with no surviving pups. b. Excludes values that were not recorded. ** Significantly different from the Group 5 value (p<0.01). 6 1.6 + CHOLESTEROL 20 67.7 + 14.1** 47.8 + 25.5** [ 19] a 53.2 + 30.6** l 14] a 62.5 + 32.4** ( 13] a 7 3 . 1 + 32.0** [ 12] a +3.3 + 31.1** [ 12] a 5 . 4 + 0-4** 5.2 + 0.6** ( 19] a 5.8 + 0.9** [ 14] a 6.7 + 1.1** ( 13 ]a 7.5 + 1.1** [ 12 ]a +2.0 + 0.9** [ 12] a 7 2 + CHOLESTEROL 18 68.6 + 20.2** 26.7 + 22.8** [ 14] a 32.6 + 27.1** l 7] a 33.9 + 29.3** [ 7] a 4 7 . 9 + 26.2** [ 5] a -24.3 + 24.1** [ 5]a 5.3 + 0.4** 4.9 + 0.5** [ 14] a 5 . 6 + 0.6** [ 7] a 5 . 8 + 0.8** [ 7] a 6.6 + 0.6** t 5] a + 1 . 0 + 0.2** [ 5] a 418-018:PAGE B-74 000148 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 21 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - SUMMARY - FI GENERATION PUPS MATERNAL DOSAGE GROUP DOSAGE MG/KG PFOS LITTERS EXAMINED (N) 1 VEHICLE CONTROL 19 TRANSIENT CLINICAL OBSERVATIONS: a COLD TO TOUCH N/N i/i NOT NURSING N/N 0/0 DECREASED MOTOR ACTIVITY N/N 0/0 GASPING N/N 0/0 LABORED BREATHING N/N 0/0 NOT NESTING N/N 0/0 8 0.4 20 1/1 1/1 9 0.8 17 10 11 12 1 1.2 1.6 19 18 17 TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS 16/1 14/1 48/3 3/3 1/1 0/0 1/1 2/2 2/2 0/0 1/1 0/0 1/1 1/1 0/0 0/0 0/0 0/0 1/1 0/0 0/0 0/0 0/0 1/1 0/0 0/0 0/0 0/0 13 2 19 4/3 5/3 0/0 0/0 0/0 0/0 PERSISTENT CLINICAL OBSERVATIONS: a TAIL MISSING FROM BASE N/N 0/0 0/0 0/0 0/0 5/1 0/0 0/0 TIP OF TAIL MISSING N/N 0/0 0/0 0/0 4/1 0/0 0/0 0/0 TIP OF TAIL BLACK N/N 2/1 0/0 2/1 0/0 0/0 0/0 0/0 STATISTICAL ANALYSES WERE RESTRICTED TO THE NUMBER OF LITTERS WITH OBSERVATIONS. N/N = TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS a. Tabulation restricted to adverse observations; all other pups appeared normal. 418-018:PAGE B-75 000149 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 21 (PAGE 2): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - SUMMARY - FI GENERATION PUPS MATERNAL DOSAGE GROUP DOSAGE MG/KG PFOS LITTERS EXAMINED (N) TRANSIENT CLINICAL OBSERVATIONS: a NOT NURSING N/N COLD TO TOUCH N/N DECREASED MOTOR ACTIVITY N/N NO MILK IN STOMACH N/N 2 MEVALONIC ACID CONTROL 18 34 1.6 + MEVALONIC ACID 18 2 + MEVALONIC ACID 19 TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS 0/0 19/3 35/5 0/0 3/1 12/4 0/0 1/1 0/0 0/0 1/1 0/0 PERSISTENT CLINICAL OBSERVATIONS: a DISTAL PORTION OF TAIL MISSING N/N 0/0 0/0 STATISTICAL ANALYSES WERE RESTRICTED TO THE NUMBER OF LITTERS WITH OBSERVATIONS. N/N = TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS a. Tabulation restricted to adverse observations; all other pups appeared normal. 1/1 418-018:PAGE B-76 000150 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 21 (PAGE 3) : CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - SUMMARY - FI GENERATION PUPS MATERNAL DOSAGE GROUP DOSAGE MG/KG PFOS LITTERS EXAMINED (N) TRANSIENT CLINICAL OBSERVATIONS: a NOT NURSING N/N COLD TO TOUCH N/N NOT NESTING N/N DECREASED MOTOR ACTIVITY N/N RIGHT EYE, EXOPHTHALMOS N/N SWOLLEN b N/N 5 CHOLESTEROL CONTROL 17 6 1.6 + CHOLESTEROL 20 7 2 + CHOLESTEROL 19 TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS 0/0 6/3 49/8** 1/1 49/5 27/6 0/0 0/0 4/2 0/0 2/1 13/2 4/2 0/0 0/0 1/1 0/0 0/0 PERSISTENT CLINICAL OBSERVATIONS: a TAIL MISSING N/N 0/0 0/0 DISTAL PORTION OF TAIL MISSING N/N 0/0 5/1 TIP OF TAIL BLACK N/N 3/1 0/0 TIP OF TAIL MISSING N/N 2/1 0/0 STATISTICAL ANALYSES WERE RESTRICTED TO THE NUMBER OF LITTERS WITH OBSERVATIONS. N/N = TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS a. Tabulation restricted to adverse observations; all other pups appeared normal. b. Located on the right forelimb and right forepaw. ** Significantly different from the Group 5 value (p<0.01). 4/1 0/0 0/0 0/0 418-018:PAGE B-77 000151 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 22 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - FI GENERATION PUPS MATERNAL DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 11 12 1 1.2 1.6 13 2 LITTERS EXAMINED N 19 20 17 19 18 17 19 TOTAL UNSCHEDULED NECROPSIES a STILLBORN FOUND DEAD N N N 5 4 i 12 8 4 8 13 17 43 22 i4 6 11 16 39 91 3 88 NO MILK IN STOMACH b N<%) 1(100.0) 1( 25.0) 3( 50.0) 7( 63.6) 5( 31.2) 26( 66.7) 37 ( 42.0) TONGUE PROTRUDED N<%) 2( 40.0) 0( 0.0)`* 0( 0.0)** 0( 0.0)** 0 ( 0 .0)** 0 ( 0 . 0 ) ** 0 ( 0.0)** SCHEDULED PUP NECROPSIES ON DAY 5 POSTPARTUM LITTERS EVALUATED N 19 20 17 18 18 13 PUPS EVALUATED N 253 274 228 253 206 112 5 43 APPEARED NORMAL LITTER INCIDENCE PUP INCIDENCE N (%} 19(100.0) N (% ) 253(100.0) 19( 95.0) 273( 99.6) 17(100.0) 228(100.0) 18(100.0) 253(100.0) 18(100.0) 206(100.0) 13(100.0) 112(100.0) 5(100.0) 43(100.0) LIVER, MEDIAN LOBE, TAN AREAS LITTER INCIDENCE N (% ) 0( PUP INCIDENCE N (% ) 0( 0.0) 0.0) M 5.0) K 0.4) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) a. Restricted to pups in which complete necropsies were performed. autolysis or cannibalization precluded evaluation. b. Analysis restricted to pups found dead and necropsied. ** Significantly different from the Group 1 value (p<0.01) . Complete necropsies were not performed on pups in which 418-018:PAGE B-78 000152 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 22 (PAGE 2): NECROPSY OBSERVATIONS SUMMARY - FI GENERATION PUPS DOSAGE GROUP DOSAGE MG/KG PFOS 2 MEVALONIC ACID CONTROL 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC ACID LITTERS EXAMINED N 18 18 19 TOTAL UNSCHEDULED NECROPSIES a STILLBORN FOUND DEAD N N N 4 2 2 60 157 62 54 155 NO MILK IN STOMACH b N(%) 1( 50.0) 35 ( 64.8) 86( 55.5) NO ORAL OPENING, NO EYES EARS DISPLACED N (% ) 1( 25.0) 0( 0.0)** 0( 0.0)** SCHEDULED PUP NECROPSIES ON DAY 5 POSTPARTUM LITTERS EVALUATED PUPS EVALUATED N N 18 231 12 94 2 3 APPEARED NORMAL LITTER INCIDENCE PUP INCIDENCE N (%) N (%) 18(100.0) 231(100.0) 12(100.0) 94(100.0) 2(100.0) 3(100.0) a. Restricted to pups in which complete necropsies were performed. autolysis or cannibalization precluded evaluation. b. Analysis restricted to pups found dead and necropsied. ** Significantly different from the Group 2 value (p<0.01). Complete necropsies were not performed on pups in which 41-8-018:PAGE B-79 000153 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 22 (PAGE 3): NECROPSY OBSERVATIONS - SUMMARY - FI GENERATION PUPS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL LITTERS EXAMINED N 17 20 19 TOTAL UNSCHEDULED NECROPSIES a N 6 48 83 STILLBORN N 2 2 2 FOUND DEAD N 4 46 81 NO MILK IN STOMACH b N (%) 1( 25.0) 33 ( 71.7)** 3 2 ( 39.5) SCHEDULED PUP NECROPSIES ON DAY 5 POSTPARTUM LITTERS EVALUATED PUPS EVALUATED N N 17 240 12 113 5 36 APPEARED NORMAL LITTER INCIDENCE PUP INCIDENCE N(%) N<%) n ( i o o .0 ) 240(100.0) 12(100.0) 113(100.0) 5(100.0) 36(100.0) a. Restricted to pups in which complete necropsies were performed. autolysis or cannibalization precluded evaluation. b. Analysis restricted to pups found dead and necropsied. ** Significantly different from the Group 5 value (pcO.Ol). Complete necropsies were not performed on pups in which 418-018:PAGE B-80 000154 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 1) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 VEHICLE CONTROL RAT tf DESCRIPTION 10901 10902 10903 10904 10905 10906 10907 10908 10909 10910 10911 10912 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 DG ( 21 ) DG ( 0) DG ( 0- 5) DG ( 12 ) DL ( 2- 5) DL ( 3 ) DS( 32- 43) DG ( 0- 2) DG ( 5- 8) DG ( 16- 21) DL ( 1- 5) DS ( 9- 18) DS ( 28- 34) DS ( 27- 39) DS ( 27- 42) DG ( 0- 21) DL ( 1- 5) DL ( 2 ) DS ( 5- 14) DS ( 23- 42) DG ( 0- 21) DL ( 1- 5) INCISORS: MISSING/BROKEN a RED PERIVAGINAL SUBSTANCE NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN a INCISORS: MISALIGNED LOCALIZED ALOPECIA UNDERSIDE LOCALIZED ALOPECIA UNDERSIDE LOCALIZED ALOPECIA NECK LOCALIZED ALOPECIA UNDERSIDE LOCALIZED ALOPECIA UNDERSIDE a LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS RIGHT FOREPAW: 2ND DIGIT SWOLLEN RIGHT FOREPAW: 2ND DIGIT MISSING RIGHT FOREPAW: 2ND DIGIT MISSING RIGHT FOREPAW: 2ND DIGIT MISSING a RED PERIVAGINAL SUBSTANCE LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS a DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION SSTOOO 418-018:PAGE B-81 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 23 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 VEHICLE CONTROL RAT ft DESCRIPTION 11511 11512 11513 11514 DS( 3- 42) DS ( 40- 42) DG ( 0- 21) DG ( 0- 21) DL( 1- 5) DL ( 1- 5) NO ADVERSE FINDINGS NO ADVERSE FINDINGS TAIL BENT LOCALIZED ALOPECIA: TAIL BENT LOCALIZED ALOPECIA: TAIL BENT a LOCALIZED ALOPECIA: NO ADVERSE FINDINGS UNDERSIDE UNDERSIDE UNDERSIDE a DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018 :PAGE B-82 00015 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 3 (PAGE 3) CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 2 MEVALONIC ACID CONTROL RAT # DESCRIPTION 10915 10916 10917 10918 10919 10920 10921 10922 10923 10924 10925 10926 10927 10928 DG ( 1) DG ( 4- 9) DG ( 13- 21) DG ( 10- 12) DG ( 17 ) DS ( 10 ) DS( 34 ) DS ( 41- 45) DG ( 0- 2) DG ( 4- 5) DG ( 10 ) DG ( 13 ) DG ( 15 ) DG ( 17 ) DG ( 17 ) DG ( 19 ) DG ( 17 ) DG ( 19 ) DG ( 15 ) DG ( 18 ) DS ( 37- 40) DS ( 43- 44) DG ( 0 ) DS ( 45 ) DG ( 5 ) DG ( 18 ) DS ( DG( DG ( DG ( DG ( DG ( DG ( DG ( 30 ) 11 ) 11 ) 12 ) 12 ) 12 ) 12 ) 12 ) DG ( 1) NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE a MOUTH: SCAB (0.1 CM IN DIAMETER) CHROMORHINORRHEA EXCESS SALIVATION EXCESS SALIVATION RIGHT EAR SWOLLEN RIGHT EAR SWOLLEN EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION RED, DRIED PERIORAL SUBSTANCE EXCESS SALIVATION MODERATE PERIORAL SUBSTANCE RED, MODERATE PERIORAL SUBSTANCE EXCESS SALIVATION CHROMORHINORRHEA RED PERIVAGINAL SUBSTANCE RED, SLIGHT PERIORAL SUBSTANCE INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA EXCESS SALIVATION SOFT OR LIQUID FECES DECREASED MOTOR ACTIVITY CHROMORHINORRHEA a CHROMODACRYORRHEA a RED PERIORAL SUBSTANCE a MORIBUND SACRIFICED; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018(PAGE B-83 000157 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 23 (PAGE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 2 MEVALONIC ACID CONTROL RAT # DESCRIPTION 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 DG ( 16 ) DS ( 33- 35) DG ( 17 ) DS ( 6) DS ( 13 ) DS ( 28- 32) DS ( 40- 44) DG ( 0- 2) DG ( 14- 17) DG ( 15 ) DG ( DG ( DG ( DG ( 17 ) 11 ) 18 ) 18 ) DG ( 12- 21) DL( 1- 5) DG ( 20 ) DS ( 6) EXCESS SALIVATION NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN EXCESS SALIVATION RALES CHROMORHINORRHEA LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS RED PERIORAL SUBSTANCE EXCESS SALIVATION EXCESS SALIVATION RED, SLIGHT, DRIED PERIORAL SUBSTANCE NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a EXCESS SALIVATION NO ADVERSE FINDINGS EXCESS SALIVATION DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018:PAGE B-84 000158 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 5) CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID RAT # DESCRIPTION 192 9 10930 10931 10932 10933 10934 10935 10936 10937 10938 10939 10940 10941 10942 11529 11530 11531 11532 11533 DS ( 40- 42) DS ( 40- 44) DG ( 0- 3) DG ( 7 ) DG ( 7 ) DG ( 9 ) DG 1 13 ) DG ( 15 ) DG ( DG ( DS ( DG ( DS ( 9- 20) 17 ) 1) 17 ) 2) DG ( 13 ) DG ( 19 ) DG ( 19- 21) DL ( 4 ) DG ( 13- 21) DG ( 18- 21) DL ( 1- 5) DL ( 1- 5) DS ( 6 ) DS ( 33- 35) DG ( DL ( DG ( DG ( DL ( 21 ) 2) 5) 16 ) 3) NO ADVERSE FINDINGS SWOLLEN SNOUT MOUTH: SCAB (0.1 CM IN DIAMETER) MOUTH: SCAB (0.1 CM IN DIAMETER) CHROMORHINORRHEA SWOLLEN SNOUT EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: NECK EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION FOUND DEAD NO ADVERSE FINDINGS EXCESS SALIVATION NO ADVERSE FINDINGS RED, SLIGHT PERIORAL SUBSTANCE INCISORS: MISSING/BROKEN a SACRIFICED DUE TO NO SURVIVING PUPS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: LIMBS a LOCALIZED ALOPECIA: UNDERSIDE a EXCESS SALIVATION INCISORS : MISSING/BROKEN NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION CHROMORHINORRHEA SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018:PAGE B-85 000159 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 6): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID RAT tt DESCRIPTION 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 DL ( 2 ) DS ( 28- 32) DS ( 14- 18) DS ( 19- 23) DG ( 12 ) DS ( 4 ) DS ( 4 ) DS ( 4- 6) DS ( 8- 12) DS ( 12 ) DS ( 12 ) DS ( 12 ) DL ( 3 ) DL ( 2 ) DG ( 13 ) DG ( 16 ) DS ( 41- 44) DG ( 0 ) DG ( 15 ) NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN MOUTH: SCAB (0.1 CM IN DIAMETER) EXCESS SALIVATION EXCESS SALIVATION LABORED BREATHING RALES RALES RED SUBSTANCE IN CAGE EXCESS SALIVATION SCANT FECES SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION RED, SLIGHT, DRIED PERIORAL SUBSTANCE INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN EXCESS SALIVATION DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION 418-018:PAGE B-86 0001*0 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PROS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 7): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID RAT # DESCRIPTION 10943 10944 10945 10946 10947 10948 10949 10950 10951 10952 10953 10954 DL ( 3) DL ( 5 ) DS ( 32 ) DS ( 34- 35) DS ( 38- 39) DS ( 41 ) DG ( 0- 1) DG( 17- 21) DS ( 34 ) DL ( 4 ) DS ( 4- 44) DS ( 37 ) DG ( 0- 21) DG ( 14- 15) DG ( 14- 16) DG ( 14- 18) DG ( 14- 21) DG ( 15- 18) DG ( 15- 19) DG ( 17 ) DG ( 17- 18) DS ( 38 ) DG ( 14 ) DG ( 8) DG ( 16 ) DL ( 2) DS( 34 ) DG ( 20- 21) DS ( 12 ) DS( 14 ) DS ( 15- 17) DS ( 16 ) DG ( 21 ) SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS a EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS TIP OF TAIL MISSING CHROMORHINORRHEA TIP OF TAIL MISSING a CHROMODACRYORRHEA CHROMORHINORRHEA RALES PTOSIS RED, SLIGHT OR MODERATE PERIORAL SUBSTANCE DEHYDRATION SCANT FECES EMACIATION NO ADVERSE FINDINGS CHROMORHINORRHEA EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION INCISORS: MISSING/BROKEN a SOFT OR LIQUID FECES CHROMORHINORRHEA INCISORS: MISSING/BROKEN SOFT OR LIQUID FECES INCISORS: MISSING/BROKEN a DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION T9T000 418-018:PAGE B-87 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 8): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID RAT # DESCRIPTION 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 DG ( 15 ) DL ( 2) DL( 2 ) DL ( 2) DS ( 11 ) DS ( 22- 27) DS ( 22- 28) DS ( 22- 38) DS ( 28- 36) DL ( 2 ) DL ( 1) DS ( 18 ) DG ( 1 ) DG ( 2 ) DG ( 10 ) DG ( 17 ) DL ( 2 ) DG ( 10 ) DL ( 4 ) DG ( 1) DG ( 12 ) DG ( 16 ) DL ( 3) DS ( 18 ) DS ( 41- 42) DG ( 0- 21) DG ( 13 ) DG ( 16 ) DL ( 1- 5) DL ( 2 ) DS ( 18 ) DG ( 16- 17) DL ( 3) DG ( 4 ) DG ( 13 ) DG ( 18 ) DL ( 3 ) EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION SWOLLEN SNOUT MOUTH: SCAB (0.2 CM IN DIAMETER) INCISORS : MISALIGNED INCISORS : MISSING/BROKEN SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION RED, SLIGHT PERIORAL SUBSTANCE EXCESS SALIVATION EXCESS SALIVATION NO ADVERSE FINDINGS RED, SLIGHT, DRIED PERIORAL SUBSTANCE SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS RED, SLIGHT PERIORAL SUBSTANCE CHROMORHINORRHEA CHROMORHINORRHEA SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION PTOSIS PTOSIS EXCESS SALIVATION RED, MODERATE PERIORAL SUBSTANCE PTOSIS SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION RED, SLIGHT PERIORAL SUBSTANCE SACRIFICED DUE TO NO SURVIVING PUPS RED, SLIGHT PERIORAL SUBSTANCE EXCESS SALIVATION EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION 418-018:PAGE B-88 000162 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 9): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID RAT # DESCRIPTION 11556 11557 DL ( DG ( DG ( DL( i) 10 ) 20 ) 1) SACRIFICED DUE TO NO SURVIVING PUPS EXCESS SALIVATION EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION 68-e3DVd-8lO"8Ii7 000163 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 23 (PAGE 10): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 5 CHOLESTEROL CONTROL RAT # DESCRIPTION 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 DG ( 6- 15) DG ( 19- 21) DS( 35- 44) DS ( 37- 44) DG ( 0- 6) DG ( 0- 21) DG ( 11- 21) DS ( DG ( DL ( DL( 39 ) 9- 20) 1- 5) 4- 5) DS ( 22- 32) DS ( 16 ) DS ( 16 ) DS ( 16 ) DS ( 16 ) DL ( 2 ) DS ( 23- 32) DG ( 19- 21) DL ( 1- 5) DS ( 27- 45) DG ( 0- 25) INCISORS: MISALIGNED NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMODACRYORRHEA a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: NECK LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: NECK a LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a LOCALIZED ALOPECIA: LIMBS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS NO ADVERSE FINDINGS DECREASED MOTOR ACTIVITY PALE IN APPEARANCE COLD TO TOUCH MORIBUND SACRIFICED RED PERIVAGINAL SUBSTANCE LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018:PAGE B-90 000164 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 11): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 5 CHOLESTEROL CONTROL RAT # DESCRIPTION 11568 11569 11570 11571 DS ( 25 ) DS( 25- 40) DS ( 42- 44) DG ( 0- 21) DL ( 1- 4) DL( ) DS ( 40- 43) DS ( 40- 45) DG ( 0- 1) DG ( 10- 21) DL( 1- 4) SWOLLEN SNOUT INCISORS : MISALIGNED INCISORS : MISALIGNED INCISORS: MISALIGNED INCISORS : MISALIGNED RED PERIVAGINAL SUBSTANCE LOCALIZED ALOPECIA: NECK LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE NO ADVERSE FINDINGS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION SSTO00 418-018:PAGE B-91 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 12) CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL RAT tt DESCRIPTION 1D971 10972 10973 10974 10975 10976 10977 10978 10979 10980 10981 10982 10983 10984 DL ( 3) DS ( 35- 42) DG ( 0- 2) DG ( 14 - 19) DG ( 14- 20) DL ( 1- 4) DL ( 2) DL ( 4 ) DS ( 9 ) DS ( 44 ) DG ( 11- 17) DG ( 18- 21) DG ( 16- 21) DS ( 40- 44) DG ( 0- 7) DG ( 1- 5) DG ( 7- 8) DS ( 33- 45) DG ( 0 - 20) DG ( 4- 20) DG ( 7- 20) DL ( 1- 2) DL ( l- 5) DL ( 1- 5) DG ( 8- 18) DL ( 3) SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN INCISORS : MISALIGNED INCISORS: MISALIGNED RED PERIVAGINAL SUBSTANCE SACRIFICED DUE TO NO SURVIVING PUPS CHROMORHINORRHEA NO ADVERSE FINDINGS CHROMORHINORRHEA CHEST : ULCERATION (DID NOT EXCEED 2.0 CM X 1.0 CM) CHEST: SCAB (2.0 CM X 0.5 CM)a LEFT FORELIMB: ULCERATION (0.5 CM X 0.2 CM)a NO ADVERSE FINDINGS LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS : MISSING/BROKEN INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA UNDERSIDE LOCALIZED ALOPECIA BACK LOCALIZED ALOPECIA BACK LOCALIZED ALOPECIA LIMBS a LOCALIZED ALOPECIA UNDERSIDE a LOCALIZED ALOPECIA UNDERSIDE SACRIFICED DUE TO NO SURVIVING PUPS DS - DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 99T000 418-018:PAGE B-92 PROTOCOL 418 -018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 13) CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL RAT # DESCRIPTION 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 DL ( DL ( 3) 3) DG ( 1- 2) DL ( 3 ) DL ( 2) DG ( 18- 21) DL ( 1- 2) DG ( 12- 19) DL ( 5) SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS CHROMODACRYORRHEA NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 23 (PAGE 14): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL RAT # DESCRIPTION 10985 10986 10987 10988 10989 10990 10991 10992 10993 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 DG ( 2- 3) DG ( 18- 20) DL ( 1- 3) DL( 3 ) DG ( 16 ) DG ( 1- 4) DG ( 20- 21) DL ( 3) DL ( 2) DG ( 18 ) DL ( 3 ) DL ( 3> DL ( 2 ) DG ( 10- 16) DG ( 12- 15) DL ( 5) DG ( 15- 20) DL ( 1- 3) DL ( 3) DL ( 2) DS( 23- 34) DG ( 2- 20) DL ( 1- 5) DL ( 5) NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA CHROMODACRYORRHEA INCISORS: MISSING/BROKEN a SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS CHROMORHINORRHEA NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS INCISORS: MISSING/BROKEN CHROMODACRYORRHEA SACRIFICED DUE TO NO SURVIVING PUPS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018 :PAGE B-94 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 15): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL RAT # DESCRIPTION 11595 11596 11597 11598 11599 DL ( i) DG ( 19- 20) DL ( 1- 3) DL( 3 ) DG ( 1- 22) DG ( 13- 22) DL ( 1- 4) DL( 1- 4) DS ( 37- 38) DL ( 3) DL ( 2 ) SACRIFICED DUE TO NO SURVIVING PUPS LOCALIZED ALOPECIA: BACK LOCALIZED ALOPECIA: BACK a SACRIFICED DUE TO NO SURVIVING PUPS LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: UNDERSIDE INCISORS: MISSING/BROKEN SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018:PAGE B-95 000169 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 16): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 8 0.4 MG/KG PFOS RAT # DESCRIPTION 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 DS ( 14- 38) DS ( 39- 44) DG ( 0- 20) DL ( 1- 5) DG ( 15 ) DG ( 15- 21) DL ( 1) DS ( 20- 27) DS ( 23- 27) DS ( 18- 42) DG ( 0- 20) DL ( 1- 5) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS FOREPAWS AND LEFT FORELIMB: SCABS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMODACRYORRHEA INCISORS: MISALIGNED NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a (EACH 0.1 CM IN DIAMETER) DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018`.PAGE B-96 00017 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 17): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 9 0.8 MG/KG PFOS RAT # DESCRIPTION 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 DS( 32- 34) DS( 18- 20) DS ( 18- 42) DS ( 27- 42) DG ( 0- 20) DG ( 0- 20) DG ( 13- 19) DL( 1- 2) DL ( 1- 5) DL( 2- 3) DL ( 2- 5) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMODACRYORRHEA INCISORS: MISALIGNED CHROMODACRYORRHEA CHROMODACRYORRHEA INCISORS: MISALIGNED INCISORS: MISSING/BROKEN INCISORS : MISALIGNED CHROMODACRYORRHEA a LACRIMATION INCISORS : MISSING/BROKEN a DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at n e c r o p s y . DL = DAY OF LACTATION 418-018:PAGE B-97 000171 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 18): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 10 1 MG/KG PFOS RAT tt DESCRIPTION 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623b 11624 11625 11626 11627 11628 11629 DG ( 14- 20) DL ( 1- 4) DG ( 13- 18) DS ( 38- 42) DG( 0- 21) DL( 1- 4) DS ( 34- 44) DG ( 0- 20) DL ( 1- 5) DG ( DL ( DL ( DG ( DG ( 5- 21) 1- 4) 3) 2) 2- 10) LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE SACRIFICED DUE TO NO SURVIVING SWOLLEN SNOUT INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. Observation confirmed at necropsy. b. Rat 11623 was missing and found on day 19 of study. 418-018:PAGE B-98 000172 PROTOCOL 418-0:8; ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 19): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 11 1.2 MG/KG PFOS RAT # DESCRIPTION 11029 11030 11031 11032 11033 11034 1103 5 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 DS ( 17- 43) DG ( 0- 20) DL( 1- 5) DG ( 11- 20) DL ( 1- 4) DG ( 11- 20) DG ( 19- 20) DG ( 19- 20) DL( 1- 4) DL ( 1- 4) DL ( 1- 4) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA LOCALIZED ALOPECIA LOCALIZED ALOPECIA NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA LOCALIZED ALOPECIA LOCALIZED ALOPECIA LOCALIZED ALOPECIA LOCALIZED ALOPECIA LOCALIZED ALOPECIA LIMBS LIMBS LIMBS a BACK BACK UNDERSIDE LIMBS BACK LIMBS UNDERSIDE BACK DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 20): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 12 1.6 MG/KG PFOS RAT # DESCRIPTION 11039 11040 11041 11042 11043 11044 11045 11046 11047 11048 11049 11050 11051 11052 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 DL ( 2 ) DG ( 5- 21) DS ( 37- 44) DG ( 0- 3) DL ( 2 ) DG ( 10- 19) DS ( 11- 44) DG ( 0- 21) DL ( 1- 5) DG ( 11- 20) DL ( 1- 5) DG ( 0- 20) DG ( 12- 17) DL ( 1- 5) DL ( 5 ) DL ( 2 ) DL ( 5 ) DG ( 11- 13) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS LOCALIZED ALOPECIA UNDERSIDE a NO ADVERSE FINDINGS INCISORS : MISSING/BROKEN INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA NECK NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA LIMBS a NO ADVERSE FINDINGS LOCALIZED ALOPECIA UNDERSIDE LOCALIZED ALOPECIA UNDERSIDE a LOCALIZED ALOPECIA LIMBS LOCALIZED ALOPECIA NECK LOCALIZED ALOPECIA LIMBS a LOCALIZED ALOPECIA NECK a SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018:PAGE B-100 000174 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 23 (PAGE 21) CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 13 2 MG/KG PFOS RAT # DESCRIPTION 11053 11054 11055 11056 11057 11058 11059 11060 11061 11062 11063 11064 11065 11066 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 DL ( DL ( 4) 2) DL ( 3 ) DL ( DL ( 3) 2) DG ( 17- 21) DG ( 17- 21) DL ( 1- 2) DL ( 1- 5) DS ( 17- 43) DG ( 0- 20) DL ( 1- 3) DL ( 3) DL ( 2 ) DG ( 12- 21) DL ( 1) DL ( 1) DL { 3) DL ( 2) DL ( DL ( DL ( 2) 2) 5) DL ( 2 ) NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: NECK LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: NECK a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS SACRIFICED DUE TO NO SURVIVING PUPS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 418-018:PAGE B-101 000175 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 1 VEHICLE CONTROL 10901 10902 10903 10904 10905 10906 10907 10908 10909 10910 10911 10912 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 DG 21 DG 21 DG 21 DG 21 DG 21 DL 5 DG 21 DG 21 DL 5 DG 21 DL 5 DG 21 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 P P P P P P P P P P P NP P P P P P P P P P P P P P P P P ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-I02 0OO17C PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 2 MEVALONIC ACID CONTROL 10915 10916 10917 10918 10919 10920 10921 10922 10923 10924 10925 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DL 5 DL 5 DL 5 DL 5 DG 21 P p p P P p P p p P p ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10926 DG 12 p MORIBUND SACRIFICED ON DAY 12 OF GESTATION (DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT). ESOPHAGUS: PERFORATION (0.2 CM X 0.3 CM). LEFT AXILLARY AREA: TISSUE DISCOLORED TAN. UTERINE CONTENTS: 14 IMPLANTATION SITES (14 LIVE EMBRYOS).b ALL OTHER TISSUES APPEARED NORMAL. 10927 10928 11515 11516 11517 11518 11519 DL 5 DG 21 DL 5 DL 5 DL 5 DL 5 DL 5 P p P p P p P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11520 DL 5 P KIDNEYS: BILATERAL, MISSHAPEN; BILATERAL, PELVIS, SLIGHT DILATION. BLADDER: WALLS THICK; CONTAINED ONE CALCULUS (0.8 CM X 0.5 CM X 0.2 C M ) . ALL OTHER TISSUES APPEARED NORMAL. 11521 11522 DL 5 DG 25 p NP ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. ' *-*'" * ^ovplnnmental ages. 418-018:PAGE B-103 000177 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 2 (CONT.) MEVALONIC ACID CONTROL 11523 11524 11525 11526 11527 11528 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 p p p P p p ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-104 000178 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 3 1.6 + MEVALONIC ACID 10929 10930 10931 10932 10933 10934 10935 DL 5 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 P P P P P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10936 DS 2 FOUND DEAD ON DAY 2 OF STUDY (DEATH OCCURRED 58 MINUTES AFTER THE 2ND DOSAGE ADMINISTRATION). THORACIC CAVITY: CONTAINED DARK RED CLOTTED MATERIAL. ALL OTHER TISSUES APPEARED NORMAL. 10937 10938 10939 10940 DG 21 DL 5 DG 25 DG 21 P P NP P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10941 DL 4 P SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10942 11529 11530 11531 DL 5 DL 5 DL 5 DG 25 P P P NP ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11532 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11533 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11534 DL 5 P ALL TISSUES APPEARED NORMAL. 11535 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-105 000179 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 5): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 3 (CONT.) 1.6 + MEVALONIC ACID 11536 11537 11538 DL 5 DL 5 DL 5 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11539 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11540 DL 5 P ALL TISSUES APPEARED NORMAL. 11541 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11542 11543 DL 5 DL 5 P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 901`9 HDVd:8l0`8Il7 Oaf PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 6): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 4 2 + MEVALONIC ACID 10943 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10944 DL 5 P SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10945 10946 10947 DG 21 DG 21 DG 21 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10948 DL 4 P SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10949 10950 10951 DG 21 DG 21 DG 21 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10952 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10953 10954 DG 21 DG 21 P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10955 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10956 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11544 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11545 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11546 DL 1 P SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-107 000181 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 7): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 4 (CONT.) 2 + !MEVALONIC ACID 11547 11548 DL 5 DG 25 P NP ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11549 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11550 DL 4 P SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11551 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11552 DL 5 P ALL TISSUES APPEARED NORMAL. 11553 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11554 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. NECROPSY OBSERVATIONS WERE NOT RECORDED 11555 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11556 DL 1 P SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11557 DL 1 P SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-108 000182 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PROS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 24 (PAGE 8): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 5 CHOLESTEROL CONTROL 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 DL 5 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DG 25 DL 5 P p P P p P p p P p p P P P P p NP P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11562 DS 16 MORIBUND SACRIFICED ON DAY 16 OF STUDY. SMALL INTESTINES: TWISTED AND CONSTRICTED AT JEJUNUM. JEJUNUM: CONTAINED DARK RED FLUID. ALL OTHER TISSUES APPEARED NORMAL. 11563 11564 11565 11566 11567 11568 11569 11570 11571 DL 5 DL 5 DL 5 DG 25 DL 5 DL 5 DL 5 DL 5 DL 5 p p P NP p P P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018-.PAGE B-109 000183 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 9); NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 6 1.6 + CHOLESTEROL 10971 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 10972 DG 21 P ALL TISSUES APPEARED NORMAL. 10973 DL 4 P SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 10974 10975 10976 10977 10978 10979 10980 10981 10982 10983 DG 21 DG 21 DG 21 DL 5 DG 21 DL 5 DG 21 DG 21 DG 21 DL 5 P P P P P P P P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10984 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11572 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11573 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11574 11575 11576 DL 5 DL 5 DL 5 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11577 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS . ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. +2,1000 418-018-.PAGE B-110 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 10): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 6 (CONT.) 1.6 + CHOLESTEROL 11578 11579 11580 DL 5 DL 5 DL 5 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11581 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 115B2 11583 11584 DL 5 DL 5 DL 5 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11585 DL 5 P SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS . ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B- l l l 000185 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 11): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 7 2 + CHOLESTEROL 10985 DG 21 p ALL TISSUES APPEARED NORMAL. 10986 DL 3 p SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10987 10988 10989 10990 10991 10992 10993 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 p p p p p p p ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10994 DL 3 p SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10995 DL 5 p ALL TISSUES APPEARED NORMAL. 10996 DL 2 p SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS STOMACH: CONTAINED PUP TISSUE. ALL OTHER TISSUES APPEARED NORMAL. 10997 10998 DL 5 DG 25 p NP ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11586 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11587 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11588 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11589 DL 5 P SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL * DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-112 000186 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 12): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 7 (CONT.) 2 + CHOLESTEROL 11590 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11591 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11592 11593 DL 5 DL 5 P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11594 DL 5 P SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS . ALL TISSUES APPEARED NORMAL. 11595 DL 1 P SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11596 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11597 DL 5 P ALL TISSUES APPEARED NORMAL. 11598 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11599 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED' GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018.'PAGE B-113 000187 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 13): NECROPSY OBSERVATIONS - INDIVIDUAL DATA Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 P P P P P P P P P P P P P P P P P P P P ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-114 000188 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 14): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 DL 5 DL 5 DL 5 DL 5 DL 5 DS 81 DL 5 DL 5 DG 25 DL 5 DG 25 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 P P P P P NP P P NP P NP P P P P P P P P P ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-115 000189 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 15): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 DL 5 DL 5 DL 5 DG 25 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 P P P NP P P P P P P P P P P P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11627 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11628 11629 DL 5 DL 5 P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-116 060190 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 16): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 11 1.2 11029 DL 5 P 11030 DS fil NP 11031 DL 5 P 11032 DL 5 P 11033 DL 5 P 11034 DL 5 P 11035 DL 5 P 11036 DL 5 p 11037 DL 5 P 11038 DL 5 P 11630 DL 5 p 11631 DG 25 NP 11632 DL 5 P 11633 DL 5 p 11634 DL 5 P 11635 DL 5 P 11636 DL 5 P 11637 DL 5 P 11638 DL 5 P 11639 DL 5 p ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-117 000191 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 17): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 12 11039 DS 81 NP 11040 DG 21 P 11041 DL 5 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11042 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11043 11044 11045 11046 11047 DG 25 DG 21 DG 21 DG 21 DG 21 NP P P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11048 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11049 11050 11051 11052 11640 11641 11642 11643 11644 DL 5 DG 21 DG 21 DG 21 DL 5 DL 5 DL 5 DL 5 DL 5 P P P P P P P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11645 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11646 11647 11648 DL 5 DL 5 DL 5 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11649 DL 5 P SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018-.PAGE B-118 000192 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 18): NECROPSY OBSERVATIONS - INDIVIDUAL DATA Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 12 (CONT.) 11650 11651 11652 11653 DL 5 DL 5 DG 25 DL 5 P P NP P ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL ALL TISSUES APPEARED NORMAL DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-119 000193 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 24 (PAGE 19): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 13 11053 DG 21 P 11054 DG 21 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11055 DL 4 P SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11056 DG 21 P ALL TISSUES APPEARED NORMAL. 11057 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11058 11059 11060 DG 21 DL 5 DG 21 P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11061 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11062 11063 DG 21 DG 21 P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11064 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11065 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11066 11654 11655 11656 DG 21 DL 5 DL 5 DL 5 P P P P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11657 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11658 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-120 000194 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 20): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 13 (CONT.) 2 11659 DL 1 P SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11660 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11661 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11662 DL 5 P ALL TISSUES APPEARED NORMAL. 11663 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11664 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11665 DL 5 p SACRIFICED ON DAY S OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11666 DG 25 NP ALL TISSUES APPEARED NORMAL. 11667 DL 2 p SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 418-018:PAGE B-12 000195 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 5 (PAGE 1) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT T O TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER ABS. WT. REL. % TBW DOSAGE GROUP 1 VEHICLE CONTROL 10901 P 10902 P 10903 P 10904 P 10905 P 10906 P 10907 P 10908 P 10909 P 10910 P 10911 P 10912 NP 10913 p 10914 P 11501 p 11502 P 11503 p 11504 P 11505 P 11506 p 11507 P 11508 P 11509 p 11510 p 11511 p 11512 P 11513 p 11514 p 405 . 389 . 436 . 420 . 401. 344 . 430 . 417 . 294 . 442 . 300. 338 . 299 . 280 . 261. 328 . 310. 347 . 319 . 347 . 336 . 328. 342 . 359. 299. 357. 279 . 327 . C c c c C D C C D c D C D D D D D D D D D D D D D D D D 14.15 12.81 14.28 15.04 14.84 15.66 14.33 14.86 11.82 14.30 15.67 12.49 11.40 11.76 12.19 13.38 12.63 14.39 11.40 17.55 14.81 12.54 13.93 14.09 12.10 11.77 11.45 12.20 3 .49 3.29 3.28 3.58 3.70 4.55 3.33 3.56 4.02 3.24 5.22 3.70 3.81 4.20 4.67 4.08 4.07 4.15 3.57 5.06 4.41 3.82 4.07 3.92 4.05 3.30 4.10 3.73 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION 418-018:PAGE B-122 000196 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 2) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER A B S . REL. W T . % TBW DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10915 P 10916 P 10917 P 10918 P 10919 P 10920 P 10921 P 10922 P 10923 P 10924 P 10925 P 10926a P 10927 P 10928 P 11515 P 11516 P 11517 P 11518 P 11519 P 11520 P 11521 P 11522 NP 11523 P 11524 P 11525 P 11526 P 11527 P 11528 P 402 . 452 . 383 . 443 . 391 . 426 . 317. 321. 296 . 301. 428 . -- 341. 408 . 282 . 341. 328 . 337. 306. 303 . 341. c c c c c c D D D D C - D C D D D D D D D 313 . 305 . 333 . 314 . 348 . 324 . D D D D D D 12.81 14.86 12.74 16.58 14.34 19.20 13.24 13.04 12.43 12.51 16.03 10.04 13.36 16.03 11.82 13.94 14.98 13.14 11.82 13.49 13.05 14.51 12.10 13.46 12.89 16.72 12.29 3.19 3.29 3 .33 3.74 3.67 4.51 4.18 4.06 4.20 4.16 3.74 -- 3.92 3.93 4.19 4.09 4.57 3.90 3.86 4.45 3.83 4.64 3.97 4.04 4.10 4.80 3.79 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION a. Dam 10926 was moribund sacrificed on day 12 of gestation; values excluded from group averages and statistical analyses. 1000 418-018:PAGE B-123 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 5 (PAGE 3) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER ABS. WT. REL. % TBW DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 p 10930 p 10931 P 10932 p 10933 P 10934 P 10935 P 10936b 10937 p 1093B P 10939 NP 10940 P 10941c P 10942 P 11529 P 11530 P 11531 NP 11532c p 11533c P 11534 P 11535c P 11536 P 11537 P 11538 P 11539c p 11540 P 11541c P 11542 p 11543 P 284 . 386 . 382 . -- 392 . 391. 363 . --383 . 316. D C c C c c c c D 459 . -- 311. 280 . 355. c - D D D -- --336 . -- 320 . 308 . 304 . -- 330 . -- 298 . 348 . D D D D D D D 13.55 15.78 14.11 a 16.00 19.10 12.81 10.59 17.21 15.82 18.76 12.38 16.18 12.71 15.07 17.59 12.63 15.30 15.27 15.63 15.01 13.25 10.86 16.51 13 .76 13.73 15.20 4.77 4.09 3.69 -- 4.08 4 .88 3.53 -- 4.49 5.01 4.09 -- 5.20 4 .54 4.24 -- -- 4.55 -- 4.88 4.87 4.36 -- 5.00 -- 4.61 4.37 A LL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION a. Value was not recorded. b. Rat 10936 was found dead on day 2 of study; values excluded from group averages and statistical analyses. c. Dam was sacrificed due to no surviving pups prior to day 5 of lactation; values excluded from group averages and statistical analyses. 418-018:PAGE B-124 000198 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 4): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER A B S . REL. W T . % TBW DOSAGE GROUP 4 10943a P -- - 14.51 10944 P 282 . D 11.92 10945 P 4 IB . C 16.51 10946 P 421. C 17.22 10947 P 10948a P 385. -- C 14.26 - 18.07 10949 P 280. c 12.70 10950 P 421 . c 16.49 10951 P 417. c 16.95 10952a P -- - 14.33 10953 P 399. c 16.16 10954 P 407 . c 16.04 10955a P 10956a P ---- - 16.30 - 16.88 11544a P -- - 13.74 11545a P --- - 18.81 11546a P --- - 14.56 11547 P V299. D 14.89 11548 NP 11549a P 11550a P 11551a P -- -- -- - 17.01 - 13.74 - 15.29 11552 P 11553a P 310. -- D 14.55 - 16.80 11554a P --- - 13.22 11555a P -- - 13.15 11556a P -- - 13.73 11557a P -- - 16.09 -- 4.23 3.98 4.09 3.70 -- 4.54 3.92 4.06 -- 4.05 3.94 -- -- -- -- -- 4 .98 -- -- -- 4.69 -- -- -- -- -- 2 MG/KG PFOS + MEVALONIC ACID ALL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . W T . = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION a. Dam was sacrificed due to no surviving pups prior to day 5 of lactation; values excluded from group averages and statistical analyses. 418-018-.PAGE B-125 000199 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 5): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER A B S . REL. W T . % TBW DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 P 10958 P 10959 P 10960 P 10961 P 10962 P 10963 P 10964 P 10965 P 10966 P 10967 P 10968 P 10969 P 10970 P 11558 P 11559 P 11560 NP 11561 p 11562a 11563 P 11564 p 11565 P 11566 NP 11567 P 11568 P 11569 P 11570 P 11571 P 316. 448 . 448. 442 . 409. 444 . 475 . 476. 386. 379. 326 . 319 . 320. 323 . 308 . 340 . D c c c c c c c c D D D D D D D 359. -- 325 . 306. 321. D D D D 347 . 312. 355 . 242 . 379 . D D D D D 12.06 14.99 14.08 15.88 12.03 12.78 14.61 14.81 13.47 15.15 12.17 12.62 11.57 15.17 13.95 15.26 14.21 b 13.87 11.71 13.18 14.07 14.46 13.75 10.88 14.48 3.82 3.34 3.14 3.59 2.94 2.88 3.08 3.11 3.49 4.00 3.73 3.96 3.62 4.70 4.53 4 .49 3.96 -- 4.27 3.83 4.10 4.05 4.63 3.87 4.50 3.82 A LL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . WT. = ORGAN WEIGHT. REL. V TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED O N DAY 5 OF LACTATION a. Rat 11562 was moribund sacrificed on day 16 of study; values excluded from group averages and statistical analyses. b. Value was not recorded. 418-018:PAGE B-126 000200 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 5 (PAGE 6) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (*) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER ABS. REL. W T . % TBW DOSAGE GROUP 6 10971a P 10972 P 10973a P 10974 P 10975 P 10976 P 10977 P 10978 P 10979 P 10980 P 10981 P 10982 P 10983 P 10984a P 11572a P 11573a P 11574 P 11575 P 11576 P 11577a P 11578 P 11579 P 11580 P 11581a P 11582 P 11583 P 11584 P 11585 P __ 455 . -- 435 . 339. 376 . 269. 415. 316 . 402 . 373 . 429. 299 . -- -- -- 284 . 335. 307 . -- 291. 325 . 319 . -- 298 . 317 . 309. 282 . _ C - c c C D C D c C c D - - D D D - D D D - D D D D 12.35 13.58 11.43 13.67 14 .34 13.80 13.75 14.48 15.66 15.36 13.37 15.56 17.08 14.82 10.33 13.88 12.14 14.93 12.96 15.41 12.51 13.83 17.42 17.70 12.69 15.15 14.66 12.93 ___ 2.98 -- 3.14 4.23 3.67 5.11 3.49 4.96 3.82 3.58 3.63 5.71 -- -- -- 4.27 4.46 4.22 -- 4 .30 4 .26 5.46 -- 4.26 4.78 4.74 4.58 1.6 MG/KG PFOS + CHOLESTEROL ALL WEIGHTS WERE RECORDED IN GRAMS (G) . ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION a. Dam was sacrificed due to no surviving pups prior to day 5 of lactation; values excluded from group averages and statistical analyses. 418-018:PAGE B-127 000201 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 7): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER ABS. REL. W T . % TBW DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10985 P 10986a P 10987 P 10988 P 10989 P 10990 P 10991 P 10992 P 10993 P 10994a P 10995 P 10996a P 10997 P 10998 NP 11586a P 11587a P 11588a P 11589 P 11590a P 11591a P 11592 P 11593 P 11594 P 11595a P 11596a P 11597 P 11598a P 11599a P 402 . -- 394 . 397. 323 . 403 . 379. 372 . 430 . -- 282. -- 261. C - c c c c c c c - D - D -- ---- 290. -- -- 297 . 262 . 299. -- -- 275 . -- -- D D D D D - 15.39 15.35 15.90 14.96 13.91 15.27 15.47 16.30 15.14 15.64 13.99 16.60 11.28 12.74 14.88 12.53 12.24 14.76 17.35 13.14 12.43 13.24 16.06 13.75 12.10 13.25 16.71 3.83 -- 4.04 3.77 4.31 3.79 4.08 4.38 3.52 -- 4.96 -- 4 .32 -- -- -- 4.22 -- -- 4.42 4.74 4.43 -- -- 4.40 -- -- ALL WEIGHTS WERE RECORDED IN GRAMS (G) . ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION a. Dam was sacrificed due to no surviving pups prior to day 5 of lactation; values excluded from group averages and statistical analyses. 418-018:PAGE B-128 000202 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 8): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER ABS. REL. W T . % TBW DOSAGE GROUP 8 0.4 MG/KG PFOS 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 P P P P P P P P P P P P P P P P P P P P 306 . 333 . 304 . 325. 333 . 322 . 325. 334 . 340 . 325 . 344 . 383 . 281. 333 . 297. 337 . 312 . 312 . 345 . 343 . D D D D D D D D D D D D D D D D D D D D 12.95 11.72 13.06 14.78 13.93 14.31 14.76 14.91 16.47 14.96 15.30 16.13 10.83 13.32 11.89 12.79 12.46 13.39 14 .17 14.65 4.23 3.52 4.30 4.55 4 .18 4 .44 4.54 4.46 4.84 4.60 4.45 4.21 3.85 4.00 4.00 3.80 3.99 4.29 4 .11 4.27 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION 418-018:PAGE B-129 000203 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 9) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER A B S . REL. W T . % TBW DOSAGE GROUP 9 0.8 MG/KG PFOS 11009 P 11010 P 11011 P 11012 P 11013 P 11014 NP 11015 P 11016 p 11017 NP 11018 P 11610 NP 11611 P 11612 p 11613 P 11614 P 11615 P 11616 P 11617 P 11618 p 11619 p 208 . 346 . 335 . 283 . 290 . D D D D D 298 . D 279 . D 324 . D 356 . 296 . 364 . 326 . 355 . 278 . 326 . 332 . 245 . D D D D D D D D D 12.16 15.03 14.14 13.00 13.78 13.64 11.15 14.81 14.16 13.88 14.51 14.95 14.09 13.27 13.45 13.79 12.04 5.85 4.34 4.22 4.59 4.75 4.58 4.00 4.57 3.98 4.69 3.99 4 .58 3.97 4.77 4 .12 4.15 4.91 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION wzooo 418-018:PAGE B-130 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 10): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (!) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER ABS. WT. REL. % TBW DOSAGE GROUP 10 1 MG/KG PFOS 11019 P 11020 P 11021 P 11022 NP 11023 P 11024 p 11025 P 11026 p 11027 p 11028 p 11620 p 11621 p 11622 p 11623 P 11624 p 11625 p 11626 P 11627a P 11628 P 11629 p 306 . 336 . 282 . D D D 281. 337 . 314 . 308 . 277 . 300 . 275 . 317. 322 . 305. 323 . 323 . 310 . -- 324 . 331. D D D D D D D D D D D D D - D D 14.75 15.26 15.71 14.01 15.93 13.15 13.34 11.86 15.70 12.79 12.48 12.46 12.68 14.63 14.49 11.80 13.73 12.33 13.14 4.82 4.54 5.57 4.98 4.73 4.19 4 .33 4.28 5.23 4.65 3.94 3.87 4.16 4.53 4.49 3.81 -- 3.80 3.97 A LL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100, P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION a. Dam was sacrificed due to no surviving pups prior to day 5 of lactation; values excluded from group averages and statistical analyses. 418-018:PAGE B-13 000205 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 11): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER A B S . REL. W T . % TBW DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 P 11030 NP 11031 P 11032 P 11033 P 11034 P 11035 P 11036 P 11037 P 11038 P 11630 P 11631 NP 11632 P 11633 P 11634 P 11635 P 11636 P 11637 P 11638 P 11639 P 304 . D 302 . 321. 273 . 296 . 274 . 338. 280. 306. 286 . D D D D D D D D D 287 . 347 . 329. 333 . 314 . 310 . 276 . 325. D D D D D D D D 16.91 13 .38 14.72 13.08 14.24 13.19 17.69 14.23 15.44 13.22 12.77 16.47 13.80 16.42 14.47 14.62 12.29 14.49 5.56 4.43 4.58 4 .79 4.81 4.81 5.23 5.08 5.04 4.62 4.45 4.75 4.19 4.93 4.61 4.72 4.45 4.46 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . W T . = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION 418-018:PAGE B-132 000206 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 12): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER A B S . REL. WT. % TBW DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 NP 11040 P 11041 P 11042a P 11043 NP 11044 p 11045 p 11046 p 11047 p 11048a p 11049 P 11050 p 11051 p 11052 p 11640 p 11641 P 11642 p 11643 P 11644 p 11645a P 11646 P 11647 p 11648 P 11649 P 11650 p 11651 P 11652 NP 11653 P 374 . 292 . -- c D - 397 . 391. 431. 384 . -- 304 . 400 . 377. 444 . 293 . 316 . 290 . 281. 288 . -- 316 . 300 . 279. 305 . 289. 352 . C C c c - D c c c D D D D D - D D D D D D 276 . D 17.98 12.95 16.60 14.06 13.30 15.09 16.38 15.53 14.72 14.87 11.94 15.61 13.66 14.02 13.57 12.41 13.88 16.08 13.89 12.62 12.08 13.96 11.67 14 .44 10.57 4.81 4.43 -- 3.54 3.40 3.50 4.26 -- 4.84 3.72 3.17 3.52 4.66 4.44 4.68 4.42 4.82 -- 4.40 4.21 4.33 4.58 4.04 4.10 3.83 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION a. Dam was sacrificed due to no surviving pups prior to day 5 of lactation; values excluded from group averages and statistical 418-018:PAGE B-13 000207 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 25 (PAGE 13): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS (%) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT LIVER ABS. WT. REL. % TBW DOSAGE GROUP 13 2 MG/KG PFOS 11053 P 11054 P 11055a P 11056 P 11057a P 11058 P 11059 P 11060 P 11061a P 11062 P 11063 P 11064a P 11065a P 11066 P 11654 P 11655 P 11656 P 11657a P 11658a P 11659a P 11660a P 11661a P 11662 P 11663a P 11664a P 11665 P 11666 NP 11667a P 386. 447 . -- 352 . -- 421. 286. 425 . -- 390 . 418 . -- 439. 272. 293 . 279. -- --- -- -- -- 284 . -- -- 289 . C c - c - c D c - c c - - c D D D - - - - - D - - D --- 14 .13 14 .45 13 .29 13.63 15.41 16.58 14.27 16.11 14.26 16.15 14.55 14.37 12.88 16.63 11.74 14.24 12.88 14.86 12.91 14.87 14.02 13.80 12.13 15.31 12.32 13.16 17.32 3.66 3.23 -- 3 .87 -- 3.94 4.99 3.79 -- 4.14 3.48 -- -- 3.79 4 .32 4 .86 4 .62 -- -- -- -- -- 4.27 -- -- 4.55 -- ALL WEIGHTS WERE RECORDED IN GRAMS (G) . A B S . WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED O N DAY 5 OF LACTATION a. Dam was sacrificed due to no surviving pups prior to day 5 of lactation,- values excluded from group averages and statistical analyses. 418-018:PAGE B-134 000208 418-018:PAGE B-13 000209 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 1): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT # DOSAGE GROUP 1 VEHICLE CONTROL 10901 10902 10903 10904 10905 10906 10907 10908 10909 10910 10911 10912 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 1150911510 11511 11512 11513 11514 206 . 227 . 205 . 219 . 198 . 214 . 211. 217 . 197. 212 . 201. 217 . 204 . 213 . 203 . 229. 219. 228 . 208 . 234 . 222 . 221 . 208 . 232 . 216 . 234 . 210 . 225 . 214 . 224 . 237. 230 . 211. 238 . 233 . 232 . 219 . 241 . 227. 242 . 219 . 224 . 203 . 244 . 221 . 236. 222 . 249 . 234 . 242 . 219 . 249 . 218 . 246 . 222 . 239. 231. 232 . 248 . 230. 217 . 250. 244 . 234 . 232 . 252 . 234 . 250 . 224 . 231. 207 . 263 . 237. 243 . 224 . 265 . 236. 260. 226 . 260 . 221 . 256. 228 . 244 . 242 . 240 . 257. 243 . 228 . 262 . 247 . 247 . 243 . 263 . 244 . 257. 227 . 227. 207 . 280 . 234 . 253 . 240 . 278 . 244 . 261. 248 . 267 . 233 . 281. 228 . 260. 248 . 242 . 264 . 260. 251. 266 . 262 . 259. 251. 266. 244 . 269. 242 . 231. 213 . 284 . 244 . 262 . 243 . 278 . 250. 260 . 252 . 278 . 234 . 288 . 231. 276. 252 . 236 . 269. 267 . 258 . 277 . 277 . 261 . 260 . 278. 252 . 283 . 251. 240. 216 . 294 . 253 . 268. 251. 282 . 262 . 270. 259. 279. 237 . 298 . 246 . 277 . 268 . 248 . 279 . 265 . 240. 285 . 266 . 263 . 258. 288 . 263 . 284 . 239. 233 . 226. 297 . 251. 269. 261. 292 . 262 . 277 . 266. 294 . 237 . 306 . 241 . 279 . 2 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. L/l PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 2): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10915 10916 10917 10918 10919 10920 10921 10922 10923 10924 10925 10926 10927 10928 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 205 . 224 . 209 . 214 . 204 . 220 . 205 . 230. 191. 215 . 209. 220 . 211. 199 . 206. 240. 215. 229. 206 . 227 . 224 . 231 . 210 . 216 . 226 . 224 . 214 . 230. 218. 250. 228 . 234 . 223 . 240 . 223 . 224 . 209. 241. 229. 233 . 229. 223 . 205 . 254 . 229 . 238 . 214 . 227. 240 . 234 . 223 . 224 . 238 . 232 . 230. 236. 218 . 266 . 239. 241. 229 . 253 . 237 . 244 . 219. 247 . 240 . 241. 242 . 218. 214 . 273 . 239. 258 . 226 . 237 . 254 . 252 . 235 . 235 . 253 . 237 . 236 . 242 . 234 . 270 . 245 . 247 . 231. 257 . 255 . 255 . 232 . 255 . 249 . 260. 247. 233 . 220 . 280. 246 . 271. 247 . 237 . 264 . 256 . 249 . 236 . 261. 241. 244 . 252 . 249 . 287 . 248 . 262 . 243 . 266 . 257. 261. 248 . 263 . 261. 271. 266 . 237 . 222 . 274 . 247. 281. 263 . 238 . 275 . 267. 249 . 239. 262 . 256 . 248. 267 . 259. 295 . 257 . 268 . 252 . 284 . 268 . 261. 256 . 274 . 258 . 277. 282 . 237 . 223 . 283 . 262 . 296. 277 . 249 . 275 . 279 . 250 . 239 . 278. 264 . 259 . 270 . 263 290 260 254 244 287 273 271 254 261 263 280 277 242 228 294 264 304 282 258 291 280 252 250 276 262 261 271 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 418-018:PAGE B-136 000210 418-018 .`PAGE B-13 000211 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 3): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT # DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 10930 10931 10932 10933 10934 10935 10936 10937 10938 10939 10940 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 202 . 217 . 202 . 207 . 193 . 217 . 212 . 213 . 204 . 212 . 202 . 214 . 204 . 211. 202 . 247. 231. 222 . 209. 232 . 217 . 221. 214 . 236. 217 . 227 . 214 . 229. 225 . 214 . 216 . 220 . 228 . 236 . 236 . 247 . 259 . 266 . 270 . 222 . 226. 233 . 235 . 241. 222 . 234 . 245. 254 . 257 . 208 . 218 . 230 . 238 . 235. 243 . 255 . 259 . 261. 261. 221. 214 . 226 . 232 . 235 . FOUND DEAD ON DAY 2 OF STUDY 227 . 237. 244 . 249. 259 . 246 . 249 . 268 . 273 . 285 . 220. 227. 231. 227. 236. 234 . 244 . 253 . 263 . 264 . 219. 223 . 231. 232 . 240 . 235. 244 . 258 . 259. 268 . 217 . 229 . 230 . 236 . 243 . 259. 275. 290. 298. 297. 237. 254 . 256 . 254 . 251 . 238. 246. 260 . 263 . 264 . 223 . 241. 244 . 246. 251. 255. 275 . 276 . 283 . 286. 230. 232. 248 . 259. 255. 238. 258 . 261. 262 . 267 . 228 . 234 . 244 . 250. 257 . 245. 262. 268. 258 . 266. 210. 210 . 212 . 211. 216. 247 . 258 . 269 . 280 . 288 . 226. 229. 244 . 248 . 249 . 238. 238. 248 . 250 . 258 . 248 . 259 . 278 . 284 . 291. 235. 276 . 238 . 243 . 234 . 267 . 233 . 263 . 281. 233 . 280 . 239. 266. 242 . 312. 259. 268 . 257. 294 . 265. 273 . 254 . 267. 221. 297 . 254 . 264 . 302 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. -J PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 26 (PAGE 4): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 10944 10945 10946 10947 1094B 10949 10950 10951 10952 10953 10954 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 11556 11557 195 . 222 . 214 . 207 . 204 . 217 . 210 . 212 . 200 . 219 . 205 . 212 . 200. 214 . 204 . 243 . 218 . 218 . 207 . 231. 212 . 231 . 214 . 232 . 221. 234 . 213 . 229. 214 . 242 . 238 . 233 . 221. 240 . 226 . 236. 228 . 232 . 224 . 228. 220. 238. 220 . 257. 234 . 239. 218 . 244 . 228 . 250. 238 . 244 . 229 . 252. 228 . 257 . 220 . 250. 242 . 242 . 231. 247 . 241. 246 . 238 . 242 . 228 . 237. 231. 235. 237 . 265 . 237 . 249. 230. 252 . 231. 254 . 251. 257 . 243 . 262 . 234 . 270 . 234 . 250 . 263 . 251 . 236 . 255 . 248 . 254 . 251. 250 . 236 . 249. 240. 254 . 243 . 284 . 254 . 262. 236. 260. 232 . 266 . 263 . 261. 254 . 273 . 237 . 278 . 246. 258 . 270 . 263 . 239. 267 . 256 . 256. 257 . 256 . 241. 263 . 237 . 270. 248 . 292. 260 . 262 , 240 . 266. 234 . 269 . 269. 262 . 248 . 279 . 242 . 281. 249 . 258 . 274 . 266 . 238 . 274 . 255 . 263 . 261. 262 . 241. 266 . 246 . 275 . 256 . 304 . 257 . 273 . 245 . 271. 241 . 275 . 275 . 263 . 254 . 284 . 250 . 291. 247 256 278 262 243 273 259 265 267 264 251 266 252 268 259 320 264 277 246 280 244 284 272 267 261 293 252 290 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. ZTZOOO 418-018-.PAGEB-138 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 5): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT tt DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 198 . 226 . 212 . 208 . 199. 220. 202. 208 . 208 . 229 . 205 . 212 . 208 . 207. 206 . 230 . 226 . 220 . 215 . 231 . 212 . 218 . 210. 230. 214 . 230 . 207 . 224 . 220. 235 . 234 . 222 . 212 . 237 . 233 . 238. 218. 247. 226 . 222 . 232 . 247. 220. 254 . 238 . 245 . 227 . 246 . 232. 230 . 224 . 246. 221. 249. 220. 244 . 233 . 257 . 236 . 219. 218 . 251. 243 . 256. 224 . 253 . 236. 235. 240. 260. 238 . 252 . 258 . 257. 225 . 264 . 247 . 230. 232 . 256 . 225 . 266 . 232 . 255 . 243 . 250 . 262 . 273 . 285 . 294 . 250. 263 . 263 . 237. 258 . 257. 226 . 236. 254 . 266 . 290. 291. 256 . 263 . 283 . 268 . 270 . 288. 241. 252 . 255. 270 . 284 . 296 . 249. 263 . 280. 238. 249 . 263 . 245 . 260. 267. 269. 296 . 290 . 244 . 247 . 251. 265. 272 . 284 . 267 . 278 . 290 . 277 . 292 . 285 . MORIBUND SACRIFICED 274 . 286 . 284 . 247 . 258 . 257. 240 . 244 . 244 . 241 . 248 . 260 . 262 . 254 . 271. 234 . 236 . 244 . 280. 290. 296 . 238 . 247 . 253 . 263 . 270 . 286 . 266 . 306 . 271. 265. 250 . 308 . 289 . 296 . 255 . 286 . 276 . 256. 256. 295 . 259. 280. 292 . 294 . ON DAY 292 . 270 . 252 . 253 . 273 . 247 . 310. 253 . 289. 308 . 16 OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 418-018:PAGE B-139 000213 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 6): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 B 15 22 29 36 42a 49 56b 63 70 77 81 RAT ft DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 10972 10973 10974 10975 10976 10977 10978 10979 10980 10981 10982 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 195 . 219 . 208. 214 . 206 . 215 . 206 . 216. 200 . 211. 209 . 212 . 207. 218. 196. 234 . 222 . 224 . 215 . 233 . 222 . 233 . 214 . 232 . 215 . 222 . 215 . 226. 211. 244 . 223 . 228 . 223 . 241. 232 . 233 . 221. 228 . 228 . 230 . 224 . 236 . 195. 244 . 234 . 236. 237 . 246. 232 . 241 . 229. 244 . 236 . 238 . 230 . 237 . 218 . 256 . 226 . 234 . 238 . 252 . 236 . 247. 238 . 228 . 234 . 228 . 237. 248 . 202 . 249. 244 . 250 . 246 . 254 . 242 . 263 . 243 . 250 . 242 . 244 . 246. 246 . 225 . 259 . 238 . 248 . 258 . 258 . 242 . 255 . 250 . 240. 249 . 244 . 235. 250. 196 . 254 . 246. 265. 260. 261. 244 . 274 . 247 . 262 . 248 . 256 . 258 . 246 . 227 . 264 . 241. 258 . 258 . 261. 242 . 263 . 254 . 241. 252 . 249. 242 . 256 . 208 . 264 . 249 . 267 . 255 . 266 . 245. 271. 260. 274 . 249. 260 . 261. 251. 232 . 274 . 242 . 264 . 255 . 266 . 243 . 260. 264 . 251. 248 . 248 . 243 . 268 . 210. 273 . 253 . 272 . 265 . 276. 246 . 288 . 258 . 272. 258 . 257 . 269 . 253 . 231. 270. 238 . 261. 263 . 263 . 244 . 266. 262 . 255 . 245. 246. 239 . 262. 208 . 276 . 255. 276 . 263 . 274 . 250 . 291. 258 . 272. 260 . 259 . 271. 254 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 418-018:PAGE B-140 000214 418-018 :PAGE B000215 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 7): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10985 10986 10987 10988 10989 10990 10991 10992 10993 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 198 . 221. 205 . 209 . 200 . 222 . 210 . 211. 210 . 212 . 207 . 214 . 197. 206. 206. 241. 217 . 222. 215. 240. 214 . 224 . 214 . 228. 222 . 226. 215 . 225. 219. 249 . 230 . 240 . 220. 235 . 222 . 232 . 224 . 242 . 222 . 226 . 218. 230 . 219. 259. 242 . 239. 222 . 248 . 236 . 224 . 235 . 241. 231. 246 . 231. 242 . 232 . 264 . 249 . 253 . 226 . 251. 234 . 236 . 242 . 255. 233 . 239. 225 . 249 . 221. 270. 253 . 251. 234. 246. 250. 222 . 242 . 253 . 230. 247. 237 . 246 . 240 . 272 . 254 . 261. 230 . 261 . 246 . 239 . 257 . 266 . 247 . 245. 227. 258 . 240 . 275 . 256 . 256. 243. 265 . 247. 227 . 254 . 252 . 238 . 252 . 247 . 251. 246 . 270 . 250 . 263 . 231. 278 . 243 . 244 . 264 . 260 . 246 . 242 . 232 . 262 . 246 . 279. 260 . 259. 242 . 276 . 256. 224 . 265 . 252 . 234 . 254 . 243 . 248 . 250. 275 . 260 . 261. 245 . 280 . 246 . 253 . 261. 265 . 246 . 251. 229. 268 . 248 . 287 . 260. 263 . 241. 281. 255 . 225 . 266. 256 . 231. 248 . 249. 254 . 256 277 268 264 235 274 254 239 275 263 248 252 234 268 249 291 260 263 254 281 258 228 270 255 239 257 256 254 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 4* PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 8): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT # DOSAGE GROUP 8 0.4 MG/KG PFOS 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 192 . 229 . 207. 209. 197. 212 . 207 . 211. 208 . 210. 213 . 247. 216 . 229 . 206 . 228 . 219 . 221. 211. 229. 210 . 251. 225. 226 . 223 . 240 . 236. 238 . 237 . 232 . 237 . 262 . 238 . 238 . 238 . 252 . 235 . 232. 235 . 240 . 223 . 264 . 238. 239 . 238. 254 . 253 . 246. 240 . 241. 261. 280 . 226 . 253 . 242 . 268 . 240. 240. 251. 253 . 237 . 270 . 249. 243 . 249. 260 . 262 . 257 . 268 . 260. 262 . 284 , 233 . 257 . 244 . 275 . 258 . 249. 260. 272 . 235 . 274 . 255. 256. 249 . 270. 263 . 270 . 275 . 275 . 264 . 290 . 234 . 258 . 257 . 284 . 254 . 24 9. 256 . 279. 242 , 282 , 256. 263 . 264. 279. 283 . 282 . 286 . 292 , 279 , 288 . 237 , 265. 268 , 294 . 258 253 . 272 , 284 . 246 . 281. 263. 252 . 269 . 287 . 286 . 277 . 288. 280. 288 . 305 . 240. 275 . 277. 299 . 269 . 264 . 287 . 288 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 418-018:PAGE B-142 000216 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.2S) TABLE 26 (PAGE 9): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 418-018:PAGE B-143 GO O' 000217 DAY 1 8 15 22 29 36 42a 49 RAT # DOSAGE GROUP 9 MG/KG PFOS 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 195 . 216 . 217 . 203 . 199 . 222 . 220 . 212 . 199 . 217 . 205 . 232 . 219. 228 . 210 . 233 . 218 . 226 . 208 . 226. 213 . 240 . 237. 222 . 218 . 235 . 239. 223 . 225 . 243 . 234 . 256. 229 . 242 . 219. 244 . 226. 238 . 230 . 246 . 222 . 250 . 253 . 233 . 225 . 254 . 252 . 229 . 248 . 256 . 248 . 265 . 230 . 260 . 234 . 253 . 240 . 244 . 240 . 254 . 225 . 253 . 267 . 236 . 228 . 265 . 259 . 236 . 256 . 258 . 254 . 288 . 239. 269. 242 . 260 . 246 . 254 . 253 . 258 . 228 . 268 . 275 . 234 . 232 . 267 . 269. 240 . 260 . 276. 243 . 290 . 244 . 276. 248 . 270. 242 . 262 . 261. 252 . 236 . 280. 277. 242 . 240. 281. 274 . 247 , 275 . 278 254 . 296 . 251. 279 . 254 , 280. 238. 266 280 . 261 235 . 258 . 288 . 242 . 240 . 287. 277 . 234 . 278 . 264 . 255 . 298 . 255 . 289. 266 . 282 . 252 . 276 . 285 . 259. 319. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 56b 63 70 77 81 322 . 326 . 316 . 317 . 322 . PROTOCOL 418 -018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 26 (PAGE 10): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 70 77 81 RAT # DOSAGE GROUP 10 1 MG/KG PFOS 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 196 . 213 . 208 . 214 . 208 . 210. 211. 211. 198 . 217 . 206 . 244 . 223 . 221. 211. 231. 216 . 220. 206 . 220 . 219 . 256 . 224 . 231. 222 . 232 . 230 . 215 . 209. 244 . 212 . 258 . 235 . 239. 224 . 249. 225 . 248 . 223 . 243 . 228 . 267 . 230 . 241 . 231. 240 . 233 . 240 . 216 . 259. 221. 264 . 232 . 247 . 232. 258 . 243 . 266 . 240. 252 . 235 . 278 . 247. 251. 240 . 264 . 244 . 249. 226 . 270 . 230. 269. 239. 246 . 247 . 270. 255 . 277 . 249. 269. 243 . 281 . 258 . 270 . 242 . 275 . 249 . 267 . 234 . 272 . 237 . 273 . 244 . 253 . 249 . 272 . 2 56 . 292 . 260 . 272 . 252 . 292 . 260 . 278. 250. 285 . 251. 281. 238 . 281. 237. 273 . 250. 258 . 251. 288 . 262 . 303 . 268. 281. 239 296 258 276 251 288 249 265 229 288 250 274 256 264 265 293 267 311 269 297 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 418-018:PAGE B-144 0021$ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 11): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 RAT # DOSAGE GROUP ii 1.2 MG/KG PFOS 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 203 . 214 . 206 . 216 . 206 . 212 . 209. 213 . 195 . 212. 202 . 237 . 221. 223 . 210. 239. 219. 223 . 208 . 228 . 228 . 251. 233 . 231. 223 . 231. 223. 249. 214 . 238 . 210. 245 . 222. 243 . 228. 254 . 231. 236 . 219. 253 . 227. 264 . 247. 250. 235 . 241. 233 . 255 . 219. 247. 224 . 263 . 239. 255 . 243 . 257. 231. 239 . 217 . 257. 230. 275 . 257 . 260. 246 . 261. 237. 267 . 234 . 254. 232 . 267. 248 . 272. 251. 257. 241. 248 . 228 . 258 . 248. 272 . 273 . 265. 254 . 268 . 249. 267 . 236 . 258 . 231. 271. 252 . 275. 269 . 273 . 249. 250 . 231. 262 . 262 . 282 . 284 . 265. 260. 278 . 253 . 276 , 248 . 263 , 241. 275 . 258 281. 269 278 255 256 231 272 238. 286. 272 . 280. 263 . 274 . 254. 281. 237. 260 . 248 . 276 . 255. 290. 275. 279. 262 . 256 . 233 . 272 . 316. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 56b 63 70 77 81 322 . 318 . 306 . 305 . 308 . 6T2000 418-018:PAGE B-145 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 1 2 ) : BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11040 11041 11042 11043 11044 11045 11046 11047 11048 11049 11050 11051 11052 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 201 . 230 . 211. 222 . 201. 215. 212 . 210. 207. 212 . 207 . 209. 208 . 219 . 206 . 232 . 221. 218. 209. 233 . 209 . 225 . 205 . 236 . 213 . 229. 213 . 225 . 229. 241. 239 . 241. 224 . 232 . 230. 241. 223 . 230. 231. 245 . 230 . 231. 212 . 262 . 241. 233 . 220. 252 . 230. 243 . 218. 259 . 228 . 250. 232 . 236. 239 . 250 . 249. 252 . 232 . 242 . 237 . 254 . 238 . 232 . 240 . 249 . 233 . 251. 232 . 273 . 243 . 238 . 225. 260. 230. 249. 219. 263 . 232 . 265 . 249. 241. 239. 258. 256. 266 . 234 . 252 . 245 . 261. 242 . 238 . 248 . 256 . 241. 261. 234 . 280. 253 . 244 . 229. 276 . 236 . 256 . 225 . 279. 229. 272 . 254 . 250 . 252 . 271. 255 . 273 . 243 . 252 . 256. 263 . 253 . 248 . 259 . 268 . 241. 269 . 240. 280 . 255 . 247 . 230 . 278 . 238 . 259. 222 . 287 . 239. 273 . 266 . 253 . 260 269 264 270 250 259 259 272 255 260 260 274 243 268 245 285 254 244 232 288 246 262 237 284 237 282 273 255 250. 265 . 265 . 273 . 244 . 255 . 252 . 261. 259. 247 . 254 . 270. 246 . 267. 252 . 291. 262 . 245. 237 . 290. 256 . 265 . 238 . 292 . 239. 280. 274 . 256. 283. 261. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 56b 288. 63 281. 70 277. 77 275. 81 272. 418-018:PAGE B-146 000220 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 13): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 IS 22 29 36 42a 49 56b 63 70 77 81 RAT ft DOSAGE GROUP 13 2 MG/KG PFOS 11053 11054 11055 11056 11057 11058 11059 11060 11061 11062 11063 11064 11065 11066 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 199. 224 . 210 . 213 . 198 . 216. 207 . 213 . 200 . 220 . 208 . 212 . 211 . 221. 207 . 241 . 221. 226 . 211. 233 . 217 . 218 . 213 . 228 . 217 . 227. 211. 224 . 213 . 246 . 228 . 225 . 215 . 241 . 228 . 232 . 228 . 230 . 232 . 235 . 221. 237. 220 . 243 . 232 . 244 . 218 . 235 . 227. 227. 235 . 243 . 228 . 239. 232 . 242 . 218 . 262 . 234 . 217. 213 . 245 . 233 . 231. 241. 230 . 245 . 243 . 235 . 249. 226 . 247 . 244 . 262 . 237. 246 . 240. 232 . 243 . 251. 237 . 243 . 239. 262 . 226 . 280 . 238 . 221. 220 . 262 . 231. 253 . 254 . 240 . 256 . 250. 249. 254 . 224 . 243 . 241. 269. 239. 256 . 242 . 240 . 257 . 260. 242 . 252. 237 . 268 . 229 . 286 . 242 . 224 . 230. 256 . 240 . 259. 257 . 246 . 260 . 257. 254 . 250. 233 . 233 . 238 . 276. 245 . 256 . 239. 243 . 261 . 266 . 241 . 251. 245 . 267 . 233 . 284 . 245 . 232 . 231. 259 . 253 . 262 . 266 . 247. 261. 264 . 256 . 260 . 237. 237 . 245 . 276 . 245 . 256 . 246. 246 . 265. 272 . 239. 254 . 248 . 282 . 230 , 298 . 247 . 214 . 228 . 263 . 243 . 252 . 266 . 250 . 261. 263 . 258 . 253 . 242 . 247 . 250. 282 . 250 . 259. 252 . 256. 273 . 272 . 242 . 253 . 253 . 287. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 418-018:PAGE B-147 000221 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 1): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT # DOSAGE GROUP 1 VEHICLE CONTROL 10901 P 10902 P 10903 P 10904 P 10905 P 10906 P 10907 P 10908 P 10909 P 10910 P 10911 P 10912 NP 10913 P 10914 P 11501 P 11502 P 11503 P 11504 P 11505 P 11506 P 11507 P 11508 P 11509 P 11510 P 11511 P 11512 P 11513 P 11514 P 263 . 250 . 284 . 274 . 244 . 299. 291. 264 . 262 . 300 . 279. 289. 279. 250 . 232 . 306. 274 . 286 . 274 . 302 . 266 . 287 . 272 . 310 . 248. 306 . 243 . 281. 264 . 258 . 293 . 283 . 264 . 305. 298 . 276 . 273 . 312 . 279. 295 . 285. 255. 248. 320. 288 . 287 . 276. 308. 284 . 294 . 284 . 302 . 255. 317. 251. 291. 275 . 265 . 302 . 290. 266 . 308 . 286 . 288 . 281. 310. 289 . 302 . 290 . 261. 242 . 330. 289. 293 . 279. 316. 290. 294 . 294 . 306 . 256. 319. 256 . 298 . 277 . 262 . 298 . 296 . 269. 303 . 306 . 283 . 285 . 317 . 293 . 310. 277. 264 . 246 . 328. 296 . 295 . 28 9. 315. 290. 293 . 293 . 299 . 261. 317. 254 . 296 . 282 . 267 . 299. 300 . 273 . 312 . 306 . 282 . 288 . 317 . 294 . 310 . 281. 266 . 252 . 324 . 295 . 295 . 287. 322 . 292 . 297 . 294 . 307 . 267 . 320 . 257 . 301. 284 . 268 . 300 . 295 . 279 . 313 . 310 . 290. 291. 318 . 296 . 318 . 280 . 267 . 248 . 326. 304 . 294 . 290. 320 . 297 . 296. 297 . 309 . 272 . 324 . 256 . 303 . 288 . 270 . 306 . 290. 279 . 318 . 311. 287 . 292 . 322 . 297. 321. 292 . 268 . 250. 325 . 305. 300 . 289. 322 . 296 . 299. 298 . 315 . 268 . 328 . 261. 300. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 7 289 . 277 . 308 . 299 . 288 . 320 . 314 . 290 . 294 . 323 . 302 . 327 . 290 . 270 . 255. 329. 305 . 302 . 296 . 328 . 301 . 304 . 295 . 317 . 272 . 333 . 263 . 306. 8 289 . 276 . 315. 294 . 291. 327 . 314 . 296 . 290 . 327. 297 . 338 . 297. 267 . 251. 331 . 305 . 307 . 292 . 329 . 305 . 307 . 299. 316 . 269. 335 . 261. 303 . 9 294 . 277 . 316 . 304 . 293 . 324 . 326 . 304 . 293 . 331. 301 . 337 . 292 . 271. 256 . 333 . 309 . 316. 297 . 330. 312 . 313 . 300 . 322 . 278 . 343 . 263 . 311. 10 300 . 283 . 321. 303 . 299 . 338 . 332 . 300. 295 . 336 . 303 . 344 . 302 . 278 . 260. 337. 315. 318 . 305. 338 . 311. 319 . 303 . 320 . 285 . 344 . 267 . 315 . 11 304 . 294 . 323 . 316 . 303 . 332 . 339. 311. 302 . 341. 307. 340 . 307. 283 . 269 . 343 . 323 . 325 . 303 . 343 . 314 . 323 . 312 . 320 . 289 . 352 . 273 . 321. 12 310 . 290 . 325 . 320 . 305 . 344 . 346 . 313 . 308 . 341. 312 . 330 . 308 . 285 . 271. 348 . 327 . 328 . 316. 353 . 323 . 325 . 319 . 328 . 292 . 353 . 275 . 326 . Z Z Z 000 418-018:PAGE B-148 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 2) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 RAT # DOSAGE GROUP 1 VEHICLE CONTROL 10901 P 10902 P 10903 P 10904 P 10905 P 10906 P 10907 P 10908 P 10909 P 10910 P 10911 P 10912 NP 10913 P 10914 P 11501 P 11502 P 11503 P 11504 P 11505 P 11506 P 11507 P 11508 P 11509 P 11510 P 11511 P 11512 P 11513 P 11514 P 312. 299. 330 . 323 . 312 . 342 . 342 . 314 . 309. 346 . 312 . 335. 320. 287. 276 . 348 . 331. 339. 313 . 356 . 333 . 331. 326 . 340 . 291. 364 . 283 . 336 . 319. 302 . 340 . 323 . 321. 351. 342 . 317. 308 . 348 . 320 . 342 . 320 . 290. 281. 355. 341. 338. 319 . 366 . 333 . 340. 332 . 340. 298 . 362 . 283 . 334 . 324 . 312 . 347 . 338 . 326 . 351. 356. 329 . 319. 354 . 330. 341. 328. 291. 282 . 367. 337. 345. 324 . 372 . 339. 348 . 331. 350 . 307. 369. 294 . 346 . 334 . 319. 359. 343 . 337. 369. 361. 336 . 321. 358 . 341. 332 . 336 . 302 . 295 . 381. 344 . 358 . 332 . 382 . 348 . 358 . 337 . 365. 312 . 377. 299. 359. 340 . 320. 367 . 357. 350. 380. 377 . 348. 331. 373 . 358 . 339 . 354 . 308 . 302 . 388. 365. 371. 345. 400 . 369. 371. 352 . 375. 321. 396 . 312 . 381. 352 . 337. 388 . 385 . 362 . 394 . 379. 363 . 349 . 388 . 374 . 340. 370. 312. 320 . 411 . 366. 385 . 355 . 421. 374 . 391. 361. 400. 331. 408 . 323 . 393 . 370 . 352 . 399. 390 . 373 . 402 . 401. 377 . 351. 399. 383 . 334 . 377. 317. 330. 425 . 376. 396 . 369. 442 . 393 . 398. 368 . 408. 350 . 426. 335 . 404 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 20 378 . 358 . 410. 401. 383 . 423 . 415 . 400 . 369. 412 . 398 . 332 . 396. 329 . 336 . 445 . 390. 414 . 385 . 452 . 408 . 412 . 377 . 426. 366 . 442 . 353 . 420 . 21 405 . 389. 436 . 420 . 401. 441. 430. 417 . 386 . 442 . 411. 338. 418 . 339. 348 . 439 . 406 . 432 . 403 . 471 . 407 . 424 . 389. 436 . 376. 447 . 356 . 416 . 418-018:PAGE B-149 000223 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10915 P 10916 P 10917 P 10918 P 10919 P 10920 P 10921 p 10922 P 10923 p 10924 P 10925 p 10926 p 10927 P 10928 p 11515 P 11516 P 11517 P 11518 p 11519 P 11520 P 11521 p 11522 NP 11523 P 11524 P 11525 p 11526 p 11527 P 11528 P 267 . 297 . 269. 281. 259. 288 . 283 . 288 . 274 . 274 . 271. 280. 319. 246 . 228 . 296 . 271. 308 . 283 . 268 . 292 . 293 . 260 . 248 . 274 . 275 . 262 . 272 . 273 . 299 . 275 . 293 . 268 . 293 . 288 . 290. 272 . 278 . 273 . 295 . 326 . 257 . 241. 311. 277. 315 . 288 . 276 . 306 . 290 . 268 . 256 . 287 . 276 . 274 . 287 . 275 . 303 . 279. 291. 276 . 291. 287 . 298. 272 . 284 . 278 . 301. 328 . 261 . 241. 315. 281. 323 . 290. 275 . 306 . 298 . 268 . 264 . 291 . 284 . 282 . 287 . 276 . 303 . 276 . 291. 273 . 300 . 284 . 294 . 274 . 289 . 282 . 301. 328 . 264 . 242 . 311. 282 . 322 . 290 . 274 . 308 . 305 . 266 . 263 . 297. 287 . 278 . 294 . 278 . 304 . 278 . 290 . 279 . 306 . 291. 295. 275 . 287 . 285 . 305. 332 . 268 . 243 . 312 . 286 . 323 . 292 . 275 . 306 . 297 . 270 . 264 . 300 . 285 . 277 . 296 . 281 . 309. 279. 291. 280. 303 . 296 . 299 . 276 . 292 . 289. 304 . 334 . 274 . 241. 308 . 287 . 324 . 294 . 272 . 314 . 295 . 274 . 264 . 301. 288 . 277 . 300 . 286 . 314 . 280 . 295 . 286 . 304 . 300 . 301 . 282 . 296 . 290 . 308 . 334 . 272 . 244 . 310 . 289 . 322 . 297 . 276 . 313 . 297 . 272 . 266 . 310 . 284 . 282 . 296 . 286 . 313 . 285 . 300 . 289. 307. 309 . 302 . 286 . 296 . 296. 308 . 337. 279 . 243 . 310 . 292 . 324 . 296 . 289 . 310 . 302 . 277 . 270 . 307. 291. 292 . 297 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 8 295 . 313 . 289. 305 . 294 . 315. 313 . 308 . 288 . 303 . 295 . 313 . 336 . 281. 250 . 321. 296 . 324 . 300 . 283 . 321. 304 . 278 . 273 . 302 . 289 . 286 . 303 . 9 295 . 318 . 290 . 312 . 301. 318 . 314 . 313 . 292 . 302 . 296 . 315 . 339 . 285 . 250 . 321 . 297 . 333 . 308 . 283 . 323 . 304 . 276 . 273 . 316 . 291. 298 . 310. 10 302 . 323 . 295 . 320. 307 . 320 . 318 . 310 . 298 . 314 . 303 . 320 . 338 . 286 . 252 . 328 . 302 . 330 . 306 . 288 . 323 . 299. 282 . 275 . 321 . 299 . 290 . 309 . 11 305 . 330 . 299 . 321. 309 . 328 . 316 . 320 . 303 . 319 . 309. 321. 340 . 293 . 260 . 332 . 309. 339 . 313 . 292 . 333 . 305 . 293 . 283 . 330 . 306 . 296 . 318 . 12 309 . 333 . 304 . 328 . 314 . 330. 319 . 310 . 306 . 321. 317. 300 . 341. 298 . 260 . 331. 312 . 339. 315. 295 . 336 . 305 . 294 . 288 . 333 . 308 . 301. 323 . 418-018:PAGE B-150 000224 418-018:PAGE B000225 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 27 (PAGE 4) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 22 23 RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10915 P 10916 P 10917 P 10918 P 10919 P 10920 P 10921 P 10922 P 10923 P 10924 P 10925 P 10926 P 10927 P 10928 P 11515 P 11516 P 11517 P 11518 P 11519 P 11520 P 11521 P 11522 NP 11523 P 11524 P 11525 P 11526 P 11527 P 11528 P 314 . 316. 328 . 335 . 335 . 340 . 344 . 352 . 365 . 376. 305 . 308. 311. 322 . 331. 334 . 343 . 350. 362 . 378 . 316 . 320 . 327 . 335 . 348 . 338 . 342 . 347 . 364 . 365 . 318. 318. 336. 344 . 358. 321. 330. 328 . 335 . 344 . 306. 308. 316. 325 . 334 . 317 . 325. 332 . 341 . 351. 314 . 323 . 334 . 341. 360. MORIBUND SACRIFICED ON DAY 12 OF GESTATION; 348. 353 . 363 . 364 . 385 . 301. 300. 309 . 318 . 331. 265 . 273 . 279. 286 . 293 . 334 . 350 . 349. 358 . 375 . 319 . 321. 331. 339. 352 . 343 . 342 . 354 . 354 . 373 . 319. 324 . 329 . 343 . 354 . 309 . 309. 322 . 330 . 348 . 336. 340 . 341. 354 . 357. 309 . 306 . 303 . 305 . 304 . 303. 306. 311 . 324 . 331. 286 . 294 . 304 . 311. 320. 341. 341. 354 . 360 . 373 . 312 . 318 . 327 . 334 . 345 . 312. 316 . 322 . 332. 347 . 326 . 330. 339. 349 . 362 . 355 . 370 . 377 . 388 . 411. 426 . 346 . 358 . 366 . 386 . 393 . 420. 345 . 362 . 375. 379 . 394 . 412 . 372 . 388. 398 . 356. 370 . 385 . 340. 349 . 362 . 359 . 375. 381. 376. 390 . 405 . DEATH WAS ATTRIBUTED TO AN 398. 412 . 433. 351. 367. 386 . 304 . 317. 332 . 390 . 407 . 427 . 368 . 374 . 386 . 380. 404 . 413 . 371. 387 . 405 . 358 . 366 . 372 . 361. 375 . 378 . 306 . 298 . 295 . 340 . 355. 376. 339 . 346. 355. 388 . 402 . 418 . 358 . 433 . 384 . 363 . 370 . 384 . 377 . 390. 408 . 402 . 452 . 383 . 443 . 391. 426 . 422. 393 . 385 . 400 . 428 . INTUBATION ACCIDENT 408 . 340 . 436 . 393 . 422 . 406 . 386 . 387 . 302 . 382 . 369. 421. 403 . 393 . 416 . 304. 304. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION LA PROTOCOL 41B-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 27 (PAGE 5) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 RAT ft DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 P 10930 P 10931 P 10932 P 10933 P 10934 P 10935 P 10936 10937 P 10938 P 10939 NP 10940 P 10941 P 10942 P 11529 P 11530 P 11531 NP 11532 P 11533 P 11534 P 11535 P 11536 P 11537 P 11538 P 11539 P 11540 P 11541 P 11542 P 11543 P 237 . 241. 249 . 250. 275 . 281. 282 . 284 . 246 . 253 . 253 . 252. 251. 254 . 261. 262 . 234 . 241. 241 . 242 . 265 . 272 . 276. 279. 239. 240. 246. 246 . FOUND DEAD ON DAY 2 OF STUDY 266 . 271. 275 . 272 . 294 . 290. 287. 285. 250. 252. 254 . 258 . 284 . 291. 294 . 297. 244 . 250. 247. 250. 282 . 280 . 279. 280. 252. 254 . 249. 249. 313 . 316. 319. 324 . 266 . 251. 260. 260. 282. 283 . 281. 284 . 255 . 264 . 264 . 259. 305. 306. 306 . 303 . 268 . 269 . 274 . 274 . 268 . 275 . 283 . 289. 262 . 265. 270. 270. 267. 274 . 271. 271. 223 . 232 . 232 . 235 . 298 . 301. 306. 308. 255 . 265 . 271 . 261 . 282 . 284 . 283 . 280 . 286 . 307 . 310 . 313 . 254 . 285 . 254 . 260. 247. 282 . 245 . 274 . 289 . 257. 297. 253 . 280. 250 . 320. 255 . 287. 271. 307 . 275 . 286. 272 . 271. 237 . 307 . 263 . 286 . 313 . 255. 280. 257 . 266 . 246 . 281. 245 . 274 . 287 . 258 . 296 . 252 . 277 . 254 . 320 . 252 . 293 . 269. 308 . 278 . 285 . 270. 271. 235. 308 . 263 . 292 . 312 . 256. 280 . 255. 270. 250 . 286. 251. 274 . 295 . 260 . 302 . 252 . 281. 250 . 327 . 263 . 289 . 276 . 309 . 278 . 280. 267 . 274 . 236. 315 . 267. 288 . 318 . 264 . 285. 260 . 269 . 249 . 287 . 247. 278 . 290 . 257 . 301. 256 . 287 . 258 . 322 . 260 . 292 . 271. 311. 280 . 288. 272 . 281. 236. 313 . 273 . 291. 320 . 263 . 293. 262 . 276 . 251. 290. 249. 283 . 299 . 253 . 308 . 261. 286. 260 . 324 . 257. 297. 268. 310. 280. 293 . 271. 279 . 239. 313 . 269. 291 . 325 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 9 10 266 . 289 . 267 . 282 . 252 . 296. 255 . 274 . 295. 274 . 283 . 250 . 293 . 258. 284 . 307 . 249 . 308 . 260 . 291. 265 . 328 . 256 . 294 . 272 . 314 . 290 . 290 . 275 . 281. 241 . 320 . 272. 299 . 328. . 289. 306. 254 . 313 . 266 . 288 . 273 . 332 . 261. 303 . 279. 328 . 289. 293 . 281. 283 . 244 . 323 . 277 . 304 . 330. 11 275 . 298 . 274 . 287. 261. 296 . 260 . 292 . 312. 258 . 325 . 268 . 296 . 273 . 341. 261 . 309 . 278 . 331. 294 . 300 . 290 . 285 . 248 . 329 . 282 . 311 . 340 . 12 282 . 304 . 281. 290. 262. 301. 267 . 300 . 320 . 252. 319 . 269. 299 . 275. 344 . 260 . 312. 284 . 336 . 302 . 310 . 290 . 297 . 247 . 340 . 289 . 313 . 341. 418-018:PAGE B-152 000226 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 6): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 RAT tf DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 P 10930 P 10931 P 10932 P 10933 P 10934 P 10935 P 10936 10937 P 10938 P 10939 NP 10940 P 10941 P 10942 P 11529 P 11530 P 11531 NP 11532 P 11533 P 11534 P 11535 P 11536 P 11537 P 11538 P 11539 P 11540 P 11541 P 11542 P 11543 P 282 . 289. 300 . 310. 308 . 307 . 321 . 320. 280 . 285. 288 . 302 . 292 . 290 . 308 . 308 . 264 . 273 . 289. 300 . 304 . 310 . 316 . 322 . 273 . 271. 287. 291. FOUND DEAD ON DAY 2 OF STUDY 294 . 304 . 309. 319. 320. 326 . 343 . 357 . 249. 253 . 254 . 258 . 317 . 331. 341. 351. 277. 278 . 280. 298 . 299 . 309. 310. 329. 280. 291. 296 . 302 . 355. 359. 367. 375. 262 . 267. 267 . 268 . 323 . 324 . 331. 344 . 290 . 298 . 301. 315. 346 . 357. 353 . 366 . 307 . 310. 316. 327 . 310. 320. 316 . 328. 296. 297 . 310. 320 . 301. 318. 319. 321. 254 . 258 . 268 . 269. 345. 347 . 354 . 368 . 294 . 292 . 300. 309. 321. 322 . 326 . 344 . 343 . 353 . 364 . 373 . 308 . 338 . 315 . 324 . 318. 332 . 301. 330 . 367. 250 . 364 . 314 . 346 . 323 . 391. 268 . 358 . 328. 384 . 350 . 343 . 339. 336 . 288 . 389. 328 . 355 . 385 . 327 . 356 . 332 . 338 . 337 . 352 . 310 . 348 . 387 . 255. 387 . 333 . 359 , 335 , 416. 274 . 377 . 345. 400. 361 356 354 347 301 400 338 368 402 342 . 365 . 347 . 356. 350. 360 . 332 . 350 . 401. 254 . 407 . 334 . 370 . 347 . 422 . 270 . 390 . 351. 416. 385 . 360 . 366. 362 . 311 . 418 . 350 . 385 . 414 . 359. 370 . 357 . 369. 362 . 370 . 344 . 364 . 416 . 258. 441. 343 . 382 . 354 . 443 . 270. 414 . 370. 427. 397 . 374 . 379 . 370 . 325 . 433 . 363 . 399. 426 . 381. 386 . 382 . 401. 392 . 391. 363 . 383 . 436 . 253 . 459 . 356 . 399 . 375 . 453 . 263 . 378 . 440 . 400 . 393 . 391. 389. 320 . 451. 369 . 415 . 452 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 22 254 . 266 . 23 259. 268 . 24 256 . 267 . 25 256 . 267 . 418-018:PAGE B-153 000227 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 7) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 P 10944 P 10945 P 10946 P 10947 P 10948 P 10949 P 10950 P 10951 P 10952 P 10953 P 10954 P 10955 P 10956 P 11544 P 11545 P 11546 P 11547 P 11548 NP 11549 P 11550 P 11551 P 11552 P 11553 P 11554 P 11555 P 11556 P 11557 P 246 . 275 . 283 . 276 . 251. 287 . 262 . 268 . 284 . 285 . 249. 270. 251. 276. 261. 312 . 260 . 284 . 254 . 277 . 250 . 280. 276 . 278 . 260 . 293 . 250 . 300 . 245. 273 . 294 . 279. 251. 285 . 255 . 269. 280. 287 . 258 . 272 . 254 . 291. 265. 321. 267 . 288 . 257. 281. 258 . 287. 280 . 280. 264 . 292 . 262. 305 . 255 . 271. 295 . 273 . 253 . 291. 254 . 275 . 282 . 288 . 254 . 276 . 257 . 282 . 267 . 331. 274 . 290 . 255 . 288. 260 . 291. 283 . 282 . 263 . 302 . 266 . 307 . 256. 267 . 294 . 283 . 254 . 296 . 256 . 278 . 275 . 287 . 246 . 278 . 259. 284 . 271. 337 . 270 . 291. 261. 288 . 262 . 292 . 282 . 281. 264 . 301. 269. 300 . 256 . 265. 292 . 278 . 255. 297 . 262 . 282 . 269 . 290 . 252 . 279. 254 . 281. 268 . 336 . 272 . 291. 261. 287 . 263 . 292 . 282 . 282 . 274 . 299 . 273 . 304 . 259 . 264 . 294 . 275 . 258 . 297 . 266. 288 . 280 . 288 . 252 . 283 . 258 . 285 . 269. 341. 273 . 290 . 269. 293 . 265 . 291. 285 . 287 . 277 . 299. 274 . 305 . 262 . 267 . 294 . 282 . 258 . 302 . 266 . 288 . 279 . 292 . 253 . 280 . 260 . 288 . 269. 342 . 277 . 292 . 265. 291. 267 . 294 . 287 . 285 . 266 . 300 . 275 . 310 . 264 . 276. 296 . 280 . 259 . 302 . 267 . 293 . 284 . 294 . 254 . 284 . 258 . 290. 268 . 344 . 270 . 299. 270 . 297 . 265 . 304 . 287 . 281. 277 . 308 . 274 . 311 . 262 . 272 . 298 . 283 . 267. 313 . 265 . 297 . 283 . 295. 255 . 285 . 266 . 298 . 269. 339. 276 . 297 . 269 . 296 . 269 . 301 . 292 . 287 . 277 . 307 . 279. 310 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 9 266 . 275 . 301. 293 . 264 . 314 . 274 . 298 . 286 . 293 . 262 . 288 . 264 . 301 . 269 . 348 . 280 . 309 . 261. 301 . 268 . 304 . 291 . 291. 275 . 316 . 281 . 317 . 10 274 . 279. 306 . 290 . 270. 322 . 276 . 306 . 295. 289. 268 . 290 . 273 . 306. 274 . 349. 288 . 308 . 260 . 308 . 277 . 312 . 298 . 293 . 277. 316 . 286 . 324 . 11 281 . 283 . 313 . 300 . 278 . 328 . 288 . 310 . 296 . 299 . 271. 299 . 272 . 310 . 279 . 357 . 288 . 319. 260. 313 . 281. 318 . 300 . 299. 287 . 322 . 294 . 333 . 12 284 . 284 . 316 . 304 . 273 . 325 . 290. 312 . 298 . 300. 275 . 293 . 274 . 309 . 287 . 362 . 292 . 326 . 259 . 313 . 285 . 321. 307 . 304 . 292 . 332 . 298 . 334 . 23000 418-018:PAGE B-154 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 8) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 p 10944 P 10945 P 10946 P 10947 P 10948 P 10949 p 10950 P 10951 p 10952 p 10953 p 10954 p 10955 p 10956 p 11544 p 11545 p 11546 p 11547 p 11548 NP 11549 P 11550 P 11551 P 11552 P 11553 P 11554 P 11555 P 11556 P 11557 P 292 . 294 . 323 . 303 . 280 . 331. 295 . 313 . 303 . 304 . 277 . 308 . 286 . 319. 296 . 370. 302 . 328. 261. 316 . 290. 329. 312 . 308 . 293 . 336 . 301. 336. 296. 297 . 325 . 303 . 284 . 338 . 269 . 319. 308 . 304 . 284 . 303 . 292 . 324 . 304. 372 . 305. 340 . 261. 322 . 298. 331. 314 . 316 . 301. 336 . 308 . 338. 300 . 301. 332 . 313 . 292 . 341 . 253 . 324 . 322 . 316. 296 . 315. 298 . 329. 310. 378. 309. 339. 260. 332 . 304 . 344 . 318 . 323 . 311. 344 . 320 . 359. 312 . 314 . 344 . 325 . 302 . 358 . 249. 334 . 335 . 326. 307 . 319. 308 . 339. 322. 386. 320. 351. 256 . 344 . 310 . 359 . 331. 339. 324 . 351. 337 . 362 . 330. 327 . 363 . 341. 312 . 367 . 260. 349. 344 . 339 . 326 . 333 . 327. 355 . 335 . 400. 337. 367. 258 . 358 . 324 . 377 . 346. 355 . 343 . 360 . 343 . 376. 346 . 347. 376 . 360 . 332 . 391. 267. 368 . 364 . 356 . 340 . 350. 341. 368 . 356 . 414 . 347. 380 . 254 . 373 . 330. 388 . 359. 374 . 353 . 385 . 361. 388 . 360. 354 . 396. 385 . 349. 405. 276. 380 . 376 . 363 . 347. 369. 358 . 379 . 372 . 423 . 356. 387. 258 . 386 . 349 . 403 . 371. 386. 371 . 397. 378 . 378 . 371. 370 . 401. 401. 360 . 423 . 278 . 398 . 385 . 374 . 364 . 384 . 376 . 394 . 391. 438 . 373 . 400 . 262 . 401. 369. 412 . 392 . 392. 372 . 415 . 383 . 386 . 382 . 415 . 421. 385 . 437. 280. 421. 417. 399. 407 . 399. 449 . 380. 418 . 259. 423 . 403 . 431. 401. 396. ALL WEIGHTS WERE RECORDED IN GRAMS (G) . P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 22 258 . 23 262 . 24 263 . 25 272 . 23000 418-018:PAGE B-155 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 27 (PAGE 9): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 RAT # DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 P 10958 P 10959 P 10960 P 10961 P 10962 P 10963 P 10964 P 10965 P 10966 P 10967 P 10968 P 10969 P 10970 P 11558 P 11559 P 11560 NP 11561 P 11562 11563 P 11564 P 11565 p 11566 NP 11567 p 11568 P 11569 P 11570 p 11571 p 266 . 265 . 274 . 276 . 283 . 317 . 323 . 330. 324 . 322 . 280 . 294 . 296 . 298 . 304 . 276 . 285 . 2 92 . 299 . 298 . 248. 257. 267. 265. 274 . 323 . 332 . 333 . 332 . 328 . 298 . 307 . 306 . 312 . 319. 311. 317 . 316. 321. 319 . 264 . 265 . 271. 274 . 278 . 303 . 308 . 316 . 318 . 320 . 307. 310. 314 . 314 . 320 . 274 . 280. 284 . 285 . 285 . 276 . 282 . 288 . 289. 291. 304 . 314 . 318. 308 . 319. 257 . 263 . 272 . 263 . 264 . 292 . 301. 309 . 319 . 318 . 310 . 304 . 306. 308 . 303 . 304. 313. 314. 317 . 321 . MORIBUND SACRIFICED ON DAY 16 OF STUDY 295. 311 . 314 . 320 . 320 . 276 . 282 . 296 . 296 . 299. 260 . 271. 272 . 275 . 280 . 284 . 289 . 286 . 292 . 296 . 276 . 289. 295 . 299 . 301. 252 . 261. 262 . 269 . 270 . 313 . 315 . 317 . 324 . 331 . 254 . 262 . 266 . 263 . 270. 294 . 309 . 318 . 326 . 323 . 281. 327 . 302 . 304 . 276 . 336 . 326 . 322 . 280 . 328 . 325 . 289 . 297 . 314 . 270 . 319. 303 . 326 . 328 . 298 . 285 . 302 . 303 . 272 . 332 . 271. 331. 250. 330 . 307 , 306 . 276 . 337 , 328 . 325 . 282 . 330 328 292 297 323 . 268 . 325 303 329. 327 300 . 287 306 . 310 . 277 . 333 , 269 . 329 263 . 334 . 311. 311. 280. 333 . 332 . 324 . 282 . 331. 326 . 294 . 298 . 323 . 275 . 327 . 307 . 338 . 329 . 300 . 292 . 305 . 316 . 277 . 336. 276 . 338 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 8 267 . 340 . 316 . 314 . 286. 337 . 342 . 330 . 288 . 336 . 330 . 299. 301. 327. 278 . 330. 306 . 343 . 339 . 308 . 294 . 308 . 320 . 284 . 344 . 277. 348 . 9 272 . 338. 315 . 320 . 290 . 341. 345. 336 . 289. 341. 332 . 303 . 309 . 334 . 277 . 336. 304 . 344 . 339. 306 . 300 . 311. 324 . 285 . 345 . 279. 342 . 10 282 . 337. 326 . 324 . 299. 342 . 345 . 341. 290 . 344 . 341. 305 . 315 . 338. 282 . 344 . 305. 352 . 341. 315 . 305 . 306 . 326. 290 . 353 . 291. 362 . 11 286 . 344 . 334 . 329 . 300 . 352 . 354 . 348 . 291. 353 . 343 . 313 . 314 . 336 . 290 . 349 . 306 . 354 . 351. 318 . 313 . 299 . 337 . 301. 357 . 298 . 359 . 12 296 . 347 . 329 . 334 . 301. 353 . 355 . 354 . 296 . 358 . 345 . 314 . 318 . 341 . 295 . 360 . 306. 352 . 354. 326 . 313 . 297. 345. 301. 370 . 298 . 368 . 418-018:PAGE B-156 000230 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 7 (PAGE 10): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 RAT # DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 P 10958 P 10959 P 10960 P 10961 P 10962 P 10963 P 10964 P 10965 P 10966 P 10967 P 10968 P 10969 P 10970 P 11558 P 11559 P 11560 NP 11561 p 11562 11563 p 11564 p 11565 p 11566 NP 11567 P 11568 P 11569 P 11570 P 11571 P 302 . 303 . 312 . 314 . 328 . 352 . 351. 361. 368 . 381. 335 . 337 . 332 . 358 . 370. 338 . 345 . 356 . 368 . 380. 300. 298 . 312. 318 . 341. 360. 358. 369. 383 . 393 . 358. 366 . 375. 3 94 . 406 . 357. 356. 366. 382 . 401. 294 . 303 . 316. 325. 337 . 359. 364 . 372 . 376 . 387. 347 . 353 . 357. 364 . 377 . 316 . 323 . 334 . 340. 353 . 319. 324. 328 . 338 . 348 . 349. 348 . 358 . 368 . 378 . 295 . 308 . 315. 322 . 343 . 359. 367. 376. 388 . 405 . 304 . 311. 309. 308. 307 . 353 . 363 . 366. 370. 386 . MORIBUND SACRIFICED ON DAY 16 OF STUDY 365. 368. 365. 375 . 379. 325 . 339. 342 . 346. 355 . 321. 325 . 336. 349 . 360 . 299. 299. 294 . 290 . 297 . 343 . 351. 355. 369 . 378 . 304 . 315 . 321. 322 . 339. 368 . 372 . 377. 393 . 409. 298 . 303 . 305. 312 . 320 , 375 . 378 . 383 . 391. 400. 340 . 398 . 394 . 390 . 359. 403 . 420 . 410 . 344 , 392 . 3 94 . 366 . 364 . 388 . 351, 419 . 309. 405 . 397 . 371. 370 . 292 . 389. 352 . 425 . 329 . 410 . 349. 415 . 412 . 407 . 373 . 419. 436 . 435 . 353 . 407 . 405 . 381. 384 . 398 . 366. 433 . 306 . 402 . 413 . 381. 381. 287 . 408 . 366 . 441. 339. 423 . 361. 426 . 427 . 417 . 385 . 422 . 456. 452 . 369. 424 . 419. 399 . 397. 413 . 371. 456 . 312 . 415. 425 . 395 . 397. 290 . 415 . 373 . 445 . 355 . 444 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 21 22 378 . 448 . 448 . 442 . 409 . 444 . 475. 476 . 386 . 434 . 430 . 413 . 429 . 383 . 462 . 312 . 434 . 448 . 405 . 402 . 291. 438 . 393 . 466 . 372 . 457 . 436 . 314 . 405 . 296 . 23 314 . 294 . 24 312 . 293 . 25 318 . 302 . 418-018:PAGE B-157 000231 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 11): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 P 10972 P 10973 P 10974 P 10975 P 10976 P 10977 P 10978 P 10979 P 10980 P 10981 P 10982 P 10983 P 10984 P 11572 P 11573 P 11574 P 11575 P 11576 P 11577 P 11578 P 11579 P 11580 P 11581 P 11582 P 11583 P 11584 P 11585 P 231. 287 . 239. 267 . 267 . 272 . 254 . 274 . 270 . 260 . 250 . 266 . 263 . 278 . 210 . 278 . 263 . 274 . 276 . 286 . 248 . 298 . 275 . 278 . 262 . 266. 274 . 257 . 232 . 294 . 246 . 278 . 256 . 277. 256 . 287 . 274 . 264 . 248 . 251. 272 . 281. 219. 281. 268. 268. 273. 291. 256. 314 . 275 . 280 . 269 . 264 . 282 . 254 . 236 . 293 . 249 . 280. 269. 280 . 256 . 284 . 282. 265 . 251. 259. 272 . 287 . 223 . 287. 266. 269 . 272 . 297. 262 . 314 . 282 . 284 . 275 . 268 . 282 . 259. 242 . 299. 250. 279 . 273 . 276 . 260. 279. 284 . 263 . 247. 260. 266 . 284 . 223. 287. 266. 285. 277 . 294 . 262 . 311. 283 . 291. 273 . 269 . 286. 264 . 245 . 300. 255 . 282 . 269 . 274 . 257 . 283 . 291. 265. 253 . 264 . 272 . 289 . 226. 289 . 268 . 291. 273 . 295. 264 . 314 . 283 . 288 . 276 . 268 . 284 . 260 . 248 . 306 . 258 . 286 . 274 . 274 . 260. 278 . 290. 270. 253 . 266 . 273 . 293 . 221. 294 . 273 . 292 . 277. 297. 268 . 319. 289. 291. 280 . 281. 289 . 262 . 248 . 306. 258 . 291. 279 . 274 . 263 . 290 . 294 . 272 . 253 . 268 . 278 . 294 . 228. 293 . 272 . 294 . 282 . 306. 270 . 320 . 292 . 292 . 286. 278 . 287 . 262 . 251. 308. 261. 293 . 280. 277 . 263 . 282 . 297. 274 . 255 . 268 . 278 . 294 . 232 . 300. 279 . 302 . 274 . 314 . 271. 323 . 299. 296 . 289 . 274 . 291. 258 . 248 . 319. 263 . 301. 282 . 279. 267 . 287 . 297. 266 . 257 . 272 . 282 . 297 . 231. 297. 279. 304 . 289. 309. 272 . 323 . 304 . 297 . 286 . 279. 296 . 265 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 9 256. 318 . 267 . 300. 283 . 279. 265 . 292 . 305 . 280 . 263 . 274 . 285 . 305 . 239 . 296 . 280 . 308 . 291. 312 . 281. 327 . 302 . 301. 292 . 292 . 292. 274 . 10 261. 320 . 273 . 311. 285 . 289. 271. 296 . 310. 282 . 267 . 281. 296 . 306 . 242 . 306. 285 . 313 . 293 . 321. 285 . 336. 309. 302 . 293 . 295 . 300. 270 . 11 268 . 330. 272 . 310 . 281. 283 . 274 . 300 . 314 . 286 . 261. 281. 297 . 315 . 246. 308 . 288 . 316 . 295 . 326 . 288 . 338 . 312 . 310 . 305 . 296 . 300 . 284 . 12 271 . 333 . 278 . 315 . 282. 283 . 284 . 301. 320. 290. 266 . 286 . 303 . 316 . 246 . 318 . 293 . 327 . 298 . 329. 294 . 348 . 327 . 306. 306 . 301. 311. 290 . z t z 000 418-018:PAGE B-158 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 12): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 22 23 24 25 RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 P 10972 P 10973 P 10974 P 10975 P 10976 P 10977 P 10978 P 10979 P 10980 P 10981 P 10982 P 10983 P 10984 P 11572 P 11573 P 11574 P 11575 P 11576 P 11577 P 11578 P 11579 P 11580 P 11581 P 11582 P 11583 P 11584 P 11585 P 270. 337 . 282 . 325 . 291 . 288 . 293 . 303 . 321. 292 . 275. 289. 302 . 320 . 252 . 322 . 296. 331. 303 . 335. 297 . 342 . 334 . 315 . 318 . 315 . 314 . 295 . 274 . 342 . 287 . 330 . 291 . 290. 292 . 306 . 327 . 296. 274 . 298. 304 . 325 . 254 . 325 . 304. 336. 312. 347 . 305 . 357. 335 . 319. 322 . 322 . 312 . 298 . 274 . 352 . 287 . 342 . 297 . 299. 296 . 323 . 338 . 313 . 288 . 310. 313. 334 . 262 . 339. 312. 349. 319. 356 . 306 . 359. 346 . 321. 329. 323 . 328 . 301. 280 . 364 . 296 . 357 . 302 . 306 . 304 . 327. 344 . 330 . 292 . 324 . 331. 346. 270. 342 . 327 . 353 . 324 . 370. 322 . 376 . 360 . 334 . 341. 332 . 341. 318 . 297 . 375 . 311. 374 . 304 . 322 . 319 . 336 . 350 . 348 . 307. 343 . 338 . 358 . 282 . 357 . 344 . 368. 335. 387 . 337 . 392 . 373 . 350 . 344 . 354 . 347. 336 . 304 . 400 . 322 . 388 . 319 . 333 . 335. 360. 370, 345 , 316. 362 , 358, 366. 294 . 375. 352 . 387 . 352 . 414 . 361 , 401. 387 . 364 . 365. 363 . 362 . 346 . 309. 417 . 332 . 407 . 326 . 348 . 352 . 367 . 386 . 355 . 326. 376 . 376 . 383 . 298 . 385. 370 . 400 . 356. 424 . 375 . 413 . 409. 377 . 372 . 381. 377. 367 . 321. 433 . 343 . 422 . 333 . 364 . 360. 392 . 400 . 374 . 344 . 392 . 391. 400. 312 . 399 . 374 . 411. 373 . 434 . 386 . 430 . 423 . 395 . 376 . 404 . 383 . 373 . 341. 455 . 372 . 435 . 339 . 376 . 379. 415 . 399. 402 . 373 . 429. 322. 396 . 389 . 412. 384 . 396. 436 . 398 . 397 . 389. 382 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 418-018:PAGE B-159 000233 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 13) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 RAT ft DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10985 P 10986 P 10987 P 10988 P 10989 P 10990 P 10991 P 10992 P 10993 P 10994 P 10995 P 10996 P 10997 P 10998 NP 11586 P 11587 p 11588 p 11589 p 11590 p 11591 p 11592 P 11593 P 11594 p 11595 P 11596 p 11597 P 11598 p 11599 P 257. 280 . 272 . 271 . 226 . 288 . 262 . 249. 282 ,, 280 . 260 . 258 . 242 . 280 . 246. 298 . 259. 269. 250. 293 . 288 . 235 . 271 . 256 . 252 . 253 . 256 . 276 . 269 . 277 . 283 . 280 . 230 . 289. 265 . 246 . 266 . 274 . 269 . 268 . 244 . 275 . 249 . 296. 259. 274 . 258 . 301. 296. 228 . 277 . 262 . 258 . 257 . 261. 276 . 269. 283 . 278 . 278 . 240 . 292 . 264 . 248 . 267 . 278 . 269. 267. 249. 272 . 255 . 310. 266. 284 . 264 . 301. 292 . 234 . 286. 268 . 257 . 258 . 270 . 275 . 271 . 278 . 278 . 273 . 248 . 290 . 264 . 255 . 283 . 276 . 268 . 267 . 247 . 278 . 252 . 304 . 267 . 275 . 263 . 296 . 293 . 242 . 284 . 269 . 255 . 258 . 270. 268 . 272 . 276 . 283 . 275 . 243 . 291. 266 . 253 . 280. 284 . 273 . 266 . 246 . 277. 253 . 304 . 267 . 279 . 261. 308 . 298 . 240 . 285 . 271. 256 . 260 . 269. 269. 272 . 282 . 279. 279. 252 . 298 . 271. 251. 285 . 281. 271. 268 . 252 . 279. 254 . 312. 275 . 277 . 264 . 307 . 296 . 244 . 275 . 275 . 259. 261. 272 . 270 . 273 . 291. 286 . 279 . 255 . 302 . 269 . 253 . 288 . 284 . 273 . 266 . 251. 269 . 260 . 310 . 277. 283 . 271. 309. 290 . 248 . 282 . 276 . 263 . 261. 278 . 274 . 270 . 292 . 286. 277 . 253 . 303 . 271. 261. 293 . 283 . 280 . 273 . 252 . 279 . 270 . 317 . 271 . 288 . 275 . 316 . 299. 245 . 287 . 273 . 263 . 263 . 276 . 281 . 278 . 294 . 286 . 281. 263 . 306 . 269 . 264 . 301. 284 . 282 . 278 . 258 . 279 . 268 . 316 . 275 . 303 . 273 . 322 . 302 . 249 . 286 . 280 . 271. 267 . 280 . 284 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 9 279 . 299. 289. 280 . 264 . 309 . 270 . 260. 300 . 290 . 286 . 275 . 262 . 270 . 276 . 326 . 279 . 300 . 280 . 324 . 297 . 260 . 290 . 284 . 271. 276 . 282 . 284 . 10 284 . 301. 293 . 291. 258 . 313 . 275. 266. 309. 293 . 291. 279 . 260. 265 . 279. 330 . 284 . 310 . 280 . 331. 297 . 261. 302 . 289. 277 . 266 . 288 . 290 . 11 288 . 305 . 296 . 288 . 262 . 320 . 278 . 274 . 316 . 295 . 289 . 282 . 272 . 272 . 283 . 331 . 296 . 308 . 294 . 334 . 305 . 260 . 303 . 292 . 274 . 274 . 292 . 296 . 12 288 . 308 . 304 . 290 . 266 . 322 . 289. 270. 324 . 297 . 292 . 284 . 273 . 271. 284 . 340 . 300 . 312 . 293 . 346 . 305 . 262 . 302 . 296 . 277 . 281 . 295 . 300 . 418-018:PAGE B-160 000234 418-018:PAGE B000235 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 14): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10985 P 10986 P 10987 P 10988 P 10989 P 10990 P 10991 P 10992 P 10993 P 10994 P 10995 P 10996 P 10997 P 10998 NP 11586 P 11587 P 11588 p 11589 P 11590 p 11591 P 11592 p 11593 P 11594 P 11595 p 11596 p 11597 P 11598 p 11599 p 290 . 311. 302 . 293. 270 . 323 . 284 . 270. 328 . 303 . 301. 284 . 273 . 271. 293 . 348 . 301. 315 . 306 . 356 . 307 . 274 . 321 . 305 . 286 . 284 . 295 . 300 . 298 . 316. 308 . 307. 274 . 328 . 289. 286 . 328 . 298 . 309. 290 . 274 . 262 . 296. 353 . 305 . 323 . 312 . 360 . 314 . 275. 317 . 298 . 293 . 288 . 299. 307 . 304 . 324 . 309. 318. 272 . 340 . 295 . 292 . 337 . 317. 314 . 307 . 290. 264 . 311. 366 . 314 . 329. 318 . 368 . 324 . 284 . 328 . 303 . 306 . 290. 313 . 317 . 314 . 348. 324 . 325 . 278 . 345 . 302 . 302 . 345 . 330 . 323 . 314 . 297 . 269. 320 . 376. 319. 346 . 336 . 377 . 336 . 288 . 341. 314 . 320 . 303 . 328 . 334 . 331. 364 . 331. 332. 282 . 359. 321. 316 . 362 . 344 . 340. 324 . 314 . 269. 333 . 394 . 337. 364 . 345 . 389 . 348 . 311 . 360. 323 . 338 . 316. 343 . 350 . 338 . 371. 351 . 355 . 294 . 366 . 334 . 325 . 372 . 364 . 366 . 347 . 330 . 269. 349. 404 . 352 . 380 . 363 . 411. 366. 322 . 378 . 332 . 355 . 331 . 359. 364 . 359. 396 . 366 . 368 . 308 . 378. 347 . 347 . 3 94 . 382 . 379 . 354 . 346 . 266. 361. 423 . 359. 397. 377 . 425 . 381. 337 . 396 . 342 . 364 . 341. 365 . 379 . 366 . 416 . 373 . 374 . 312 . 387 . 361. 360 . 404 . 394 . 386 . 365 . 348. 265 . 368 . 432 . 375. 410 . 389 . 446 . 401. 354 . 408 . 357 . 375 . 355 . 376 . 392 . 402 . 431. 394 . 397. 323 . 403 . 379. 372 . 430 . 424 . 396 . 364 . 272 . 375 . 423 . 418 . 421 . 360. 368 . 416 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 22 277 . 366. 23 275 . 24 274 . 25 283 . CT\ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 15): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT (t DOSAGE GROUP 8 0.4 MG/KG PFOS 10999 P 11000 P 11001 P 11002 P 11003 P 11004 P 11005 P 11006 P 11007 P 11008 P 11600 P 11601 P 11602 P 11603 P 11604 P 11605 P 11606 P 11607 P 11608 P 11609 P 250 . 298 . 273 . 281 . 284 . 281. 291 . 273 . 298 . 301. 284 . 300 . 242 . 282 . 278 . 305. 274 . 256 . 291. 289. 259 . 300 . 279 . 281. 294 . 292. 298 . 290 . 305 . 317 . 297 . 325 . 254 . 291. 278 . 314 . 279 . 267 . 305. 300 . 265 . 302 . 280 . 283 . 296 . 299. 301. 290 . 319. 315. 306. 324 . 252 . 298 . 276. 319. 280 . 270. 314 . 303 . 267 . 306 . 280 . 288 . 300 . 300. 307. 291. 320 . 315. 303 . 327 . 259. 299 . 283 . 317 . 279. 270 . 306 . 308 . 267. 308 . 281. 287 . 301. 307. 307 . 300. 330. 318. 304 . 328 . 262 . 301. 285 . 323 . 285 . 272 . 315. 311. 276 , 316 . 282 . 296 , 306. 314 . 313 . 306 , 330. 316 , 310 336 , 264 . 305 . 288 . 330 . 288 . 275 . 316 . 314 . 280. 317 . 283 . 295 . 304 . 315. 318 . 302 . 334 . 323 . 319. 336. 267. 309 . 290 . 329 . 291. 275 . 322 . 316 . ALL WEIGHTS WERE RECORDED IN GRAMS (G), P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 7 282 . 323 . 281. 304 . 304 . 316. 318 . 308 . 336. 322 . 315. 331. 268 . 314 . 294 . 330 . 293 . 283 . 324 . 322 . 8 288 . 328 . 292 . 303 . 310. 315 . 325. 312 . 339 . 316 . 319. 336. 273 . 314 . 299. 337. 295 . 280 . 327 . 327 . 9 291. 334 . 296 . 304 . 312 . 313 . 328 . 313 . 343 . 317 . 320 . 344 . 279 . 318 . 295 . 344 . 300 . 289 . 326 . 331. 10 297 . 331. 298 . 314 . 307 . 324 . 327 . 319. 348 . 328 . 328 . 350. 281. 322 . 301. 341 . 298 . 292 . 330 . 338 . 11 303 . 338 . 303 . 307 . 312 . 337 . 330. 321 . 352 . 332. 334 . 355 . 285 . 329 . 306 . 346. 309 . 303 . 342 . 339 . 12 307 . 344 . 304 . 314 . 323 . 340. 337 . 325 . 357 . 333 . 341. 363 . 292 . 332 . 320 . 352 . 312 . 310 . 344 . 345 . 418-018:PAGE B-162 000236 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 16) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 22 23 24 25 RAT # DOSAGE GROUP 8 0.4 MG/KG PFOS 10999 P 11000 P 11001 P 11002 P 11003 P 11004 P 11005 P 11006 P 11007 P 11008 P 11600 P 11601 P 11602 P 11603 P 11604 P 11605 P 11606 P 11607 P 11608 P 11609 P 310 . 351. 306 . 316 . 322. 345 . 338 . 330 . 355 . 331. 343 . 366 . 294 . 338 . 313 . 357 . 321. 306. 348 . 350. 316. 351. 312 . 327 . 320. 347. 338. 343 . 360. 329. 349. 376 . 296. 339 . 321. 364 . 321. 318 . 363 . 361. 327 . 361. 323 . 337. 348. 358. 343 . 348 . 377. 341. 357. 385. 303 . 351. 326. 370. 325. 330. 363 . 369. 331. 367. 336 . 349. 372 . 365. 349. 354 . 392 . 356 . 367. 400. 307 . 362 . 333 . 379. 332 . 346 . 364 . 379 . 339 . 380. 349. 361. 380. 375. 364 . 368. 405. 373 . 381. 412 . 325 . 374 . 345 . 393 . 345 . 354 . 387 . 398. 359 . 397. 368 . 369. 398 . 390 376 379, 427 , 378 . 397 . 424 . 332, 388 . 363 . 405 , 352 . 370 . 390. 411. 368 . 399 . 384 . 386. 402 . 406. 385 . 395 . 428 . 401. 403 . 435 . 336 . 409 . 374 . 426 . 366 . 380. 405 . 428 . 379. 422 . 392 . 403 . 416 . 422 . 397 . 417. 444 . 403 . 423 . 449. 352 . 430 . 386 . 442 . 380 . 386 . 419 . 444 . 389 428 401 414 434 412 430 459 423 438 469 350 399 457 395 409 429 446 ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 418-018:PAGE B-163 000237 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 17): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT # DOSAGE GROUP 9 0.8 MG/KG PFOS 11009 P 11010 P 11011 P 11012 P 11013 P 11014 NP 11015 P 11016 P 11017 NP 11018 P 11610 NP 11611 P 11612 P 11613 P 11614 P 11615 P 11616 P 11617 P 11618 P 11619 P 238 . 252 . 258 . 288 . 296 . 299 . 293 . 296 . 310 . 252 , 258 . 265 . 251. 251. 256 . MATING NOT CONFIRMED 296 . 296 . 298 . 236 . 252 . 253 . 284 . 290 . 298 . 296 . 308 . 312 . 248 . 248 . 251. 304 . 325. 318 . 258 . 264 . 269. 283 . 295 . 303 . 277 . 278. 277 . 293 . 300. 301. 254 . 258 . 256 . 275 . 280. 281. 296. 305. 312. 268 . 265 . 267. 255 . 307. 308. 266 . 259 . 301. 253 . 296. 307. 256. 316. 276. 303 . 285 . 300 . 252 . 279 . 306 . 267 . 254 . 305 . 312 . 262 . 264 . 293 . 256 . 307 . 309 . 260 . 323 . 270 . 306 . 286 . 304 . 261. 286 . 310. 271. 253 . 303 . 313 . 262 . 266 . 326 . 261. 308 . 308 . 283 . 326 . 274 . 309. 293 . 296 . 271. 285 . 314 . 277. 258 . 306. 310. 265 . 269. 290. 268 . 303 . 313 . 272 . 324 . 278 . 315 . 285 . 304 . 263 . 285 . 319 . 275 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 7 259. 311. 313 . 269. 271 . 294 . 267 . 307 . 310 . 279 . 329 . 283 . 315 . 283 . 304 . 268 . 293 . 321. 282 . 8 267. 309. 322 . 271. 270 . 301. 266 . 316 . 311. 279. 327 . 283 . 327. 296 . 306 . 274 . 297 . 316 . 275. 9 268 . 321. 319. 279. 277 . 304 . 268 . 319. 319. 280 . 332 . 286 . 324 . 294 . 312 . 273 . 306 . 322 . 277 . 10 268 . 333 . 307. 279. 283 . 306 . 271. 319 . 318 . 284 . 344 . 289. 333 . 304 . 316 . 286 . 310 . 326 . 286 . 11 272 . 336. 332 . 288 . 282 . 314 . 282 . 320 . 322 . 290 . 347 . 295 . 334 . 305 . 322 . 285 . 310 . 331. 290 . 12 272 . 345 . 338 . 291. 289. 312 . 282 . 321. 326 . 289. 349. 301. 341. 309 . 329 . 287 . 314 . 340 . 295 . 418-018:PAGE B-164 000238 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 18) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 418-018:PAGE B-165 CO o 000239 PREGNANCY STATUS DAY 13 14 15 16 17 18 19 RAT It DOSAGE GROUP 9 MG/KG PFOS 11009 P 11010 P 11011 P 11012 P 11013 P 11014 NP 11015 P 11016 p 11017 NP 11018 P 11610 NP 11611 p 11612 P 11613 P 11614 p 11615 P 11616 p 11617 P 11618 P 11619 P 274 . 273 . 284 . 346 . 353 . 361 . 342 . 349 . 356. 293 . 295 . 312 . 289 . 291. 302 . MATING NOT CONFIRMED 316 . 326 . 333 . 286. 288 . 296. 310. 309. 307. 332. 338 . 347 . 278 . 280. 283 . 362. 364 . 371. 304 . 310 . 316. 344 . 357 . 364 . 315. 330. 338. 333 . 340. 339. 301. 301. 324 . 321. 325 . 334 . 340 . 348 . 356. 295 . 310. 318 . 301. 377 . 370. 320 . 313 . 342 , 302 . 303 . 356 . 293 . 378 . 328 . 377. 350. 357. 330. 346 . 365. 324 . 309 . 393 . 384 . 335. 325 . 356 . 322 . 297 . 358 . 298 . 399 . 335. 388 . 366 . 365 . 348 . 365 . 378 . 344 . 328 403 . 398 344 , 337 371. 339. 302 . 368 300 . 416 . 354 . 403 . 388 . 383 , 359. 379 . 385. 356 , 330 . 417 . 426 . 366 . 349. 388 . 349 . 298 . 377 . 299 . 424 . 367. 416 . 403 . 395 . 370 . 396 . 392 . 374 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 20 339 . 422 . 412 . 378 . 363 . 406 . 364 . 288 . 394 . 299 . 435 . 376. 430 . 409. 408 . 380 . 418 . 405 . 387. 21 363 437 391 378 418 288 411 306 397 449 414 433 420 391 22 23 24 25 289. 339 . 306. 296 . 312 . 294 . 308 . 298 297 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 19): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT # DOSAGE GROUP 10 1 MG/KG PFOS 11019 P 11020 P 11021 P 11022 NP 11023 P 11024 P 11025 P 11026 P 11027 P 11028 P 11620 P 11621 P 11622 P 11623 P 11624 P 11625 P 11626 P 11627 P 11628 P 11629 P 277 . 296 . 254 . 304 . 259. 285 . 260. 289. 236 . 289. 252 . 280 . 254 . 275 . 262 . 289. 275 . 320. 266 . 292 . 278 . 309. 265 . 304 . 268 . 300. 265 . 295 . 245 . 302. 259. 290 . 261. 276 . 272 . 306 . 275 . 326 . 275 . 303 . 279. 315 . 269. 304 . 270. 317. 269 . 296 . 249. 305 . 259 . 298 . 267 . 280 . 277 . 306 . 276 . 333 . 282 . 299 . 282 . 311. 273 . 308 . 268 . 314 . 273 . 301. 253 . 303 . 263 . 300 . 269. 278 . 280. 307. 279 . 324 . 282 . 304 . 283 . 308. 278 . 312 . 274 . 297. 276 . 300 . 257 . 304 . 267 . 300 . 274 . 271 . 282 . 317 . 280 . 323 . 280. 305 . 284 . 309. 280. 316 . 273 . 323 . 277 . 302 . 261. 304 . 266 . 299. 275 . 284 . 285 . 311 . 285 . 327 . 282 . 305 . 287 . 313 . 280. 312 . 278 . 317. 282 . 302 . 260 . 297. 264 . 301. 278 . 290 . 278 . 314 . 284 . 329 . 287. 309 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 7 291. 314 . 288. 318 . 277 . 322 . 281. 301. 253 . 307 . 266. 305 . 282 . 292 . 279. 314 . 287 . 334 . 289 . 309 . 8 285 . 319 . 289 . 318 . 282 . 308 . 285 . 303 . 272 . 306 . 269 . 305 . 284 . 291. 287. 322 . 293 . 326 . 298 . 315 . 9 292 . 319 . 292 . 316 . 284 . 320. 289. 310. 2 72 . 307. 269. 307 . 290. 295 . 288 . 322 . 298 . 331 . 304 . 314 . 10 297 . 320 . 293 . 318 . 290 . 321. 295 . 314 . 274 . 311. 271 . 317 . 291. 305 . 297. 322 . 300 . 339 . 302 . 316 . 11 298 . 328 . 302 . 317 . 294 . 328 . 298 . 319 . 282 . 319. 275 . 323 . 299 . 302 . 313 . 328 . 310 . 346 . 301. 326 . 12 301. 333 . 301 . 310 . 296 . 329. 295 . 320 . 286 . 323 . 280 . 321. 297 . 306 . 316 . 343 . 311. 347 . 305. 325 . 418-018 (PAGE B-166 000240 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 20): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 RAT # DOSAGE GROUP 10 1 MG/KG PFOS 11019 P 11020 P 11021 P 11022 NP 11023 P 11024 P 11025 P 11026 P 11027 P 11028 P 11620 P 11621 P 11622 P 11623 P 11624 P 11625 P 11626 P 11627 P 11628 P 11629 P 310. 334 . 308 . 307 . 302 . 338. 306 . 324 . 290 . 332 . 286 . 328 . 308. 316 . 315. 343 . 310 . 350. 319. 329 . 313 . 333 . 316 . 309. 310. 344 . 310. 332 . 300. 339. 293 . 334 . 317 . 324 . 325. 352 . 326 . 366. 330. 341. 320 . 347 . 322 . 306 . 324 . 354 . 310 . 332 . 306 . 351. 304 . 341. 327 . 323 . 330. 360 . 328 . 372 . 333 . 345 . 331 . 357. 338 . 302 . 336 . 358 . 330. 345 . 316. 361. 317 . 350 . 334 . 344 . 341. 374 . 339. 380 . 346. 354 . 346. 366 . 358 . 299. 352. 373 . 333 . 358 . 331. 378. 319. 369. 349. 359 . 354 . 388 . 353 . 389. 366. 360 . 365 . 384 . 372 . 299. 370. 379 . 349. 374 . 343 . 400 . 342 . 385. 364 . 364 . 369. 408 . 364 . 408 . 376 . 378 . 380 . 396 . 387 . 300 . 378 . 388 . 364 . 386 . 360 . 414 . 355 . 405 . 382. 385 . 390 . 423 . 372 . 391. 397 . 394 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 20 398 . 419 . 402 . 303 . 386. 404 . 381. 402 . 377 . 436 . 369. 421. 396 . 393 . 404 . 444 . 377 . 381. 414 . 400 . 21 433 . 412 . 304 . 420 . 399. 411 . 381. 402 . 412 . 454 . 395 . 370 . 414 . 22 310 . 403 . 23 307. 24 25 311 . 309 . 418-018:PAGE B-167 000241 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 7 (PAGE 21) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT # DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 P 11030 NP 11031 P 11032 P 11033 P 11034 p 11035 P 11036 P 11037 p 11038 P 11630 P 11631 NP 11632 P 11633 P 11634 P 11635 P 11636 P 11637 P 11638 P 11639 P 262 . 274 . 270 . MATING NOT CONFIRMED 291. 295 . 296 . 277 . 283 . 288 . 272 . 272 . 280 . 281. 286 . 287 . 259. 268 . 262 . 296 . 296 . 298 . 233 . 243 . 249 . 268 . 277 . 276 . 253 . 260 . 258 . 287 . 279. 273 . 254 . 259. 260. 301. 303 . 305 . 276 . 292 . 293 . 282. 295 . 300 . 256 . 267. 278 . 258 . 268 . 270. 237 . 243 . 245 . 272 . 277. 282 . 253 . 302 . 288. 274 . 286 . 266 . 297 . 247. 280 . 261. 262 . 256 . 310 . 294 . 299. 277 . 273 . 247 . 285 . 259. 298 . 290 . 278 . 283 . 270 . 295 . 248 . 284 . 258 . 273 . 267. 311. 294 . 304 . 276 . 271. 246 . 283 . 277 , 304 . 291, 275 . 284 . 278 . 303 . 250 . 282 . 261, 278 , 267 . 307 . 292 . 300. 280 . 276 . 252 . 285 . 275 . 310 . 293 . 279. 288 . 278 . 305. 256. 288 . 263 . 284 . 267 . 315. 299 . 302 . 278 . 275. 253 . 280 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 7 276 . 311. 294 . 280. 292 . 276 . 314 . 255 . 295 . 262 . 287 . 268 . 315 . 299 . 308 . 287 . 275 . 255 . 288 . 8 276 . 314 . 297 . 286 . 293 . 280 . 315 . 260 . 297 . 268 . 294 . 265 . 328 . 304 . 313 . 285 . 280 . 261. 289 . 9 282 . 314 . 297. 285 . 296. 272 . 319 . 266 . 292 . 266 . 307 . 276 . 322 . 312 . 313 . 292 . 285 . 259. 297 . 10 283 . 316. 305 . 286 . 301. 275 . 323 . 269. 297 . 273 . 305 . 276 . 327 . 318 . 321 . 300. 284 . 262 . 300 . 11 301 . 326 . 309. 292 . 302 . 286 . 328 . 276 . 310 . 274 . 301 . 284 . 336 . 326 . 321. 306. 291. 270 . 302 . 12 293 . 328 . 313 . 299 . 309 . 291. 335 . 278 . 305 . 279 . 314 . 291. 337 . 325 . 356 . 307 . 294 . 273 . 309 . 418-018:PAGE B-168 000242 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 22) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 RAT # DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 P 11030 NP 11031 P 11032 P 11033 P 11034 P 11035 P 11036 P 11037 P 11038 P 11630 P 11631 NP 11632 P 11633 P 11634 P 11635 P 11636 P 11637 P 11638 P 11639 P 301. 300. 307. MATING NOT CONFIRMED 324 . 331. 339. 315 . 322 . 329. 304 . 308 . 314 . 312 . 319. 327 . 298 . 294 . 311. 342 . 345 . 362 . 282 . 291. 302 . 313 . 315. 329. 276 . 286. 287 . 302 . 305 . 297 . 296. 303 . 304 . 341. 346. 356 . 328 . 337. 340 . 328 . 339. 344 . 319. 328 . 317 . 298 . 312 . 312 . 277 . 290 . 290 . 314 . 326 . 334 . 324 . 344 . 343 . 329. 333 . 311. 368 . 308 . 338. 297 . 303 . 316 . 373 . 353 . 353 . 340 . 324 . 304 . 342 . 348. 357 . 367. 335 . 349. 324 . 385 . 329. 352. 304 . 290 . 324 . 384 . 361. 368 . 366 . 340 . 318 . 352 . 361. 366 . 371. 354 . 366 . 344 . 404 . 348 , 372 . 313 . 292 , 332 , 406 . 375 : 378 . 382 358 . 332 366 380 . 362 . 379. 370 . 371. 354 . 410 . 352 . 378 . 324 . 292 . 344 . 420 . 389 . 396 . 407 . 361. 343 . 377 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 20 392 . 380 . 389. 383 . 389. 368 . 432 . 367 . 402 . 328 . 286 . 353 . 428 . 401 . 412 . 415 . 381 . 352 . 396 . 21 410 . 409. 3 94 . 401. 366 . 446 . 374 . 408 . 327 . 287 . 361. 401 . 431 . 433 . 394 . 363 . 22 290. 23 288. 24 291. 25 300. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 23) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 NP 11040 p 11041 P 11042 P 11043 NP 11044 p 11045 P 11046 p 11047 P 11048 p 11049 P 11050 p 11051 P 11052 P 11640 p 11641 p 11642 p 11643 p 11644 p 11645 p 11646 p 11647 p 11648 P 11649 P 11650 P 11651 P 11652 NP 11653 P MATING NOT CONFIRMED 271 . 280 . 276. 275 . 277 . 282 . 280 . 282 . 287 . 251. 246 . 244 . 269 . 273 . 276 . 260. 266 . 271 . 274 . 279 . 279. 261. 269 . 269. 260. 269 . 271. 267 . 270. 283 . 268 . 268 . 275. 245. 248. 246 . 277. 288 . 287 . 259. 267. 266. 294 . 303 . 303 . 265 . 269. 271. 257 . 250 . 257. 236. 243 . 248 . 297 . 300 . 301. 252 . 261. 264 . 266 . 278 . 274 . 252 . 249 . 252 . 294 . 298 . 302 . 261. 263 . 267 . 297 . 303 . 303 . 290 . 300 . 300. 258 . 262 . 265 . 273 . 282 . 283 . 252 . 273 . 270 . 283 . 268 . 270 . 281. 278 . 250. 288 . 264 . 303 . 268. 257. 253 . 303 . 262 . 275 . 255 . 298 . 268 . 312 . 305 . 267. 266 . 284 . 281. 252 . 269 . 271. 282 . 273 . 270. 285. 276. 256. 294 . 264 . 293 . 275 . 261. 249 . 311. 265 . 284 . 250. 302 . 268 . 310. 309. 274 . 264 , 279 . 286 . 245 , 273 , 273 . 282 . 275 276 281. 278 255. 300 . 260 . 298 276 , 261. 251. 316 . 267, 284 . 257 . 304 . 268 . 310 . 308 . 274 . 278 . 286 . 285 . 240 . 274 . 274 . 290 . 280. 278 . 286. 282 . 256 . 295. 266 . 299. 273 . 271. 252 . 318 . 267. 273 . 258 . 308 . 271. 315 . 317 . 271 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 7 282 . 295 . 291 . 243 . 274 . 276. 291. 279 . 278 . 291. 282 . 256. 294 . 272 . 306 . 279 . 270. 263 . 319. 275 . 281. 263 . 306 . 273 . 322 . 312 . 273 . 8 287 . 293 . 293 . 243 . 275 . 278 . 296 . 276. 281. 291. 286 . 264 . 303 . 266. 310. 280 . 278 . 264 . 320. 279. 287 . 265 . 308 . 275 . 317 . 323 . 281. 9 285 . 294 . 292 . 238 . 277. 281. 300 . 280 . 287 . 293 . 287. 263 . 308 . 268 . 313 . 284 . 284 . 263 . 327. 275 . 293 . 269. 313 . 263 . 333 . 308 . 278 . 10 290 . 298 . 299 . 240 . 285 . 282 . 313 . 287. 294 . 296. 290. 270. 311. 273 . 319. 287. 282 . 273 . 334 . 283 . 295. 272 . 315 . 274 . 327. 303 . 278 . 11 281. 300 . 303 . 238. 294 . 281. 308 . 292 . 296 . 304 . 297. 271. 316 . 279 . 323 . 284 . 285 . 276 . 337 . 287. 293 . 272 . 319 . 284 . 336. 306 . 292 . 12 285 . 303 . 303 . 245 . 297 . 286 . 310. 293 . 299. 306 . 293 . 273 . 320 . 281. 323 . 288 . 292 . 278. 343 . 287. 300 . 270 . 328. 290 . 340 . 303 . 289 . 418-018:PAGE B-170 000244 000245 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 24) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 NP 11040 P 11041 P 11042 P 11043 NP 11044 P 11045 P 11046 P 11047 P 11048 P 11049 P 11050 P 11051 P 11052 P 11640 P 11641 P 11642 P 11643 P 11644 P 11645 P 11646 P 11647 P 11648 P 11649 P 11650 P 11651 P 11652 NP 11653 P MATING NOT CONFIRMED 288 . 297 . 296. 313 . 313 . 331. 313 . 310 . 319. 251. 245 . 240 . 300 . 298 . 308 . 290 . 296 . 312 . 323 . 324 . 336 . 293 . 292 . 310. 303 . 314 . 321. 304 . 312 . 319. 302 . 305 . 317. 276 . 285 . 292 . 320. 325. 340. 294 . 304 . 302 . 332 . 334 . 345 . 296. 305. 314 . 295 . 300. 305 . 282 . 290. 297. 347. 354 . 364 . 290 . 306 . 311. 310. 318. 321. 278 . 294 . 303 . 333 . 334 . 343 . 291. 297 . 305 . 342 . 352 . 357 . 302 . 303 . 307. 293 . 300 . 310 . 312 . 340. 330. 245 . 317 . 324 . 346 . 318 . 327. 330. 323 . 302. 346 . 308 . 349 . 326 . 313 . 307 . 386. 323 . 332 . 317 . 353 . 310 . 362 . 304 . 316 . 326. 358 . 346 . 250. 335 . 340 . 364 . 326. 333 . 330 . 337. 308 . 364 . 328 . 369. 342 . 328 . 318. 398 . 341. 347 . 339. 375. 329 . 371 . 306 . 333 . 340 370 , 359. 249 . 357 , 356 364 344 . 340 . 344 . 355 , 325 386 , 346 . 384 . 366 . 348 , 340. 410 . 352 . 363 . 342 . 391. 342 . 390 . 302 . 348 . 350. 391. 375 . 244 . 368 . 370 . 388 . 356. 354 . 353 . 375. 345 . 400 . 360. 393 . 382. 353 . 355. 428 . 369. 368. 353 . 407. 356 . 410 . 300 . 367 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 20 361. 416 . 394 . 233 . 380 . 376 . 404 . 365 . 368. 353 . 382 . 350. 421. 368 . 409. 386 . 373 . 366 . 441. 380 . 383 . 357 . 423 . 3 74 . 425 . 304 . 373 . 21 374 . 244 . 397 . 391. 431. 384 . 379 . 358 . 400 . 377 . 444 . 396. 421 . 383 . 450 . 384 . 375 . 428 . 385 . 429 . 304 . 380 . 22 242 . 308 . 23 244 . 310. 24 243 . 308 . 25 249 . 313 . -aHOVd:8lO-8t^ -J PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 25): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 i 2 3 4 5 6 RAT # DOSAGE GROUP 13 2 MG/KG PFOS 11053 P 11054 P 11055 P 11056 P 11057 P 11058 P 11059 P 11060 P 11061 P 11062 P 11063 P 11064 P 11065 P 11066 P 11654 P 11655 P 11656 P 11657 P 11658 P 11659 P 11660 P 11661 P 11662 P 11663 P 11664 P 11665 P 11666 NP 11667 P 237 . 303 . 256 . 222 . 230 . 265 . 251 . 266 . 267 . 249 . 268 . 285 . 261. 261. 244 , 246 . 247. 284 . 251. 259 . 263 . 256 . 272 . 268 . 238 . 262 . 265. 301. 249. 298 . 262 . 219. 235 , 267 . 251. 269 . 276 . 248 . 278. 286. 271. 262 . 248 . 260 . 256 . 294 . 256 . 308. 259. 258 . 280 . 275 . 248 . 266 . 264 . 294 . 238 . 308 . 266 . 232 . 240 . 268 . 257. 272 . 276 . 257 . 277. 291 . 270. 268 . 248 . 264 . 259. 294 . 261. 267. 262 . 262 . 278 . 279 . 248 . 269 . 264 . 299. 239. 305 . 266 . 234 . 246. 271. 256 . 270 . 273 . 262 . 274 . 295 . 267. 274 . 253 . 264 . 262 . 295 . 264 . 263 . 267 . 263 . 278 . 282 . 249 . 265 . 268 . 299 . 240 . 308 . 267 . 236 . 245 . 270 . 258. 270 . 282 . 260 . 273 . 296 . 273 . 275 . 250 . 264 . 265 . 290 . 264 . 265 . 263 . 270 . 282 . 286 . 255 . 270 . 272 . 296 . 248 . 306 . 269 . 237 . 256 . 272 . 255. 269. 282 . 266 . 277 . 302 . 274 . 271. 253 . 268 . 265. 299. 268 . 269 . 271. 268 . 285 . 280 . 250 . 276 . 274 . 303 . 247 . 315 . 271. 245 . 250. 275 . 263 . 271. 272 . 261. 276 . 299. 273 . 278 . 254 . 270 . 270 . 296 . 271. 271. 261. 271 . 286 . 289. 254 . 274 . 278 . 305 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 7 248 . 316. 277 . 246 . 256 . 278 . 261 . 273 . 282 . 268 . 281. 302 . 275 . 278 . 263 . 277 . 269. 302 . 269 . 273 . 269. 271 . 294 . 295 . 254 . 278 . 284 . 310 . 8 249 . 322 . 277 . 244 . 258 . 274 . 259. 284 . 288 . 268 . 282 . 304 . 280 . 280 . 262 . 278 . 276 . 308 . 276 . 273 . 271. 282 . 284 . 299 . 257 . 283 . 275 . 308 . 9 255 . 323 . 275 . 246 . 261. 286 . 268 . 284 . 290 . 273 . 282 . 308 . 280 . 285 . 269 . 277 . 276 . 313 . 272 . 275 . 275 . 279 . 290 . 298 . 266. 287 . 271 . 308 . 10 256 . 331. 283 . 249 . 264 . 282 . 272 . 288 . 294 . 275 . 285. 309 . 285 . 289 . 281. 282 . 282 . 314 . 281. 279. 282 . 287 . 295 . 306 . 270 . 293 . 271 . 315 . 11 265 . 338 . 290 . 255 . 264 . 291. 277 . 291 . 299 . 277 . 292 . 315 . 286 . 302 . 284 . 291. 283 . 330 . 282 . 282 . 287 . 290 . 297 . 308 . 267 . 298 . 268 . 321. 12 263 . 337 . 290 . 259 . 270 . 291. 282 . 292 . 307 . 282 . 296 . 315 . 288 . 303 . 288. 292 . 290 . 331 . 285 . 291. 289 . 293 . 303 . 316 . 272 . 300 . 266 . 325 . 418-018:PAGE B-172 00024 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 27 (PAGE 26) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 13 14 15 16 17 18 19 RAT # DOSAGE GROUP 13 2 MG/KG PFOS 11053 P 11054 P 11055 P 11056 P 11057 P 11058 P 11059 P 11060 P 11061 P 11062 P 11063 P 11064 P 11065 P 11066 P 11654 P 11655 P 11656 P 11657 P 11658 P 11659 P 11660 P 11661 P 11662 P 11663 P 11664 P 11665 P 11666 NP 11667 P 271 . 343 . 299. 265 . 278 . 300 . 285 . 301. 305 . 286. 301. 321. 295 . 308 . 288 . 304 . 294 . 332 . 292 . 286 . 296 . 294 . 308 . 318 . 274 . 308 , 263 . 328 . 276 . 335 . 300. 259. 276 . 299. 290. 305 . 315 . 291. 301. 322 . 301. 318 . 285 . 304 . 300. 333 . 292 . 294 . 306 . 305 . 315. 325 . 280 . 314 . 267. 337 . 285 . 355 . 314. 275 . 285 . 313 . 301. 317 . 324 . 301. 314 . 332. 311. 333 . 299. 315. 312. 349 . 296. 299 . 314 . 314 . 321. 329. 287 . .325 . 270 . 340. 299 . 356 . 325 . 280. 298 . 328 . 310 . 327 . 336. 313 . 324 . 337 . 321. 342. 307 . 328. 326 . 360. 303 . 302 . 324 . 327. 332 . 343 . 299. 336 . 265 . 350. 316. 370 . 342 . 295 . 312 . 346 . 325 . 352 . 358 . 317 . 341. 346 . 327. 354 . 323 . 349 . 328. 368 . 312 . 316 . 342 . 336 . 345 . 356 . 312 . 352 . 263 . 367. 329 . 384 . 365 . 306 . 328 . 369 . 342 . 363 . 365 . 333 . 358 . 361. 324 . 374 . 335 . 360 . 343 . 384 . 328 . 340 . 366 . 350. 358 . 373 . 323 . 364 . 267 . 381 . 344 . 404 . 376 . 327 . 334 . 371. 349. 374 . 383 . 349 . 378 . 373 . 346. 383 . 354 . 346. 366 . 406 . 344 . 355 . 365 . 368 . 366 . 395. 337. 380. 266 . 398 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 20 366. 425 . 388 . 336. 345 . 389 . 370 . 392 . 394 . 366 . 389. 385 . 359. 408. 375 . 371. 370 . 418 . 352 . 358 . 379 . 377 . 382 . 409 . 344 . 397. 261. 416 . 21 386 . 447 . 425 . 352 . 357 . 421. 384 . 425 . 431. 390. 418 . 408 . 382 . 439. 377 . 394 . 353 . 362 . 362 . 405 . 266 . 430 . 22 266 . 23 264 . 24 266 . 25 274 . 418-018:PAGE B-173 000247 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 1): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 1 10906 10909 10911 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 332. 277. 286 . 288 . 261. 250 . 319. 308 . 307 . 298 . 339 . 316. 319. 312 . 318. 270. 342. 277 . 306 . 344 . 294 . 300 . 299 . 280 . 261. 328 . 310. 347 . 319. 347 . 336. 328 . 342 . 359. 299. 357 . 279 . 327. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION VEHICLE CONTROL 418-018:PAGE B-174 000248 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 2): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10921 10922 10923 10924 10926 10927 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 306 . 317. 295 . 321. 277 . 296. 310 . 301. MORIBUND SACRIFICED ON DAY 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 327 . 341. 247 . 282 . 311. 341 . 282 . 328 . 337. 337. 306. 306 . 265. 303 . 333 . 341. NOT PREGNANT 274 . 313 . 275 . 305 . 322 . 333 . 302 . 314 . 317. 348. 301. 324 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-175 000249 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 3): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 3 3 .6 MG/KG PFOS + MEVALONIC ACID 10929 10936 10938 10939 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 275 . 284 . FOUND DEAD ON DAY 2 OF STUDY 314 . 316 . NOT PREGNANT 261. SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 290. 311 . 265 . 280 . 330 . 355 . NOT PREGNANT 320 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 292. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 343 . 336. 310 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 307 . 320. 284 . 308. 288 . 304 . 255 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 354 . 330. 290 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 312 . 298 . 336 . 348 . ALL WEIGHTS WERE RECORDED IN GRAMS (G), DAY = DAY OF LACTATION 418-018:PAGE B-176 000250 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 4): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 10944 10948 10952 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 11556 11557 279, SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 290 . 282. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING 322 . SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 299 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 272 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 302 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 305 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 348. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 281 . SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 300. 299. NOT PREGNANT 310. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 290. SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 326 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 296. 310 . 291. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 272 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 322 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 310. SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018 :PAGE B-177 000251 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 5) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 5 10957 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 288 . 316. 330. 379. 312 . 326 . 297 . 319. 297 . 320 . 318 . 323 . 276. 308 . 325 . 340. NOT PREGNANT 327 . 359. MORIBUND SACRIFICED ON DAY 16 OF STUDY 333. 325. 309. 306 . 291. 321. NOT PREGNANT 330 . 347. 289 . 312 . 340. 355 . 282 . 242 . 366. 379. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION CHOLESTEROL CONTROL 418-018(PAGE B-178 000252 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 6) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 10973 10977 10979 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 273 . 271. 262 . 285. 289. 309. 246 . 316 . 280 . 308 . 287. 322 . 280. 352 . 324 . 306 . 312 . 304 . 275 . 294 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 269 . 316 . 299. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 284 . 335. 307. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 291. 325 . 319. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 298. 317. 309. 282. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-179 000253 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 7): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10986 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 309. SACRIFICED ON DAY 3 OF 317 . SACRIFICED ON DAY 3 OF 277 . 282 . 291 . SACRIFICED ON DAY 2 OF 250 . 261. NOT PREGNANT 279. SACRIFICED ON DAY 3 OF 340. SACRIFICED ON DAY 3 OF 312. SACRIFICED ON DAY 2 OF 303 . 290. SACRIFICED ON 290 . SACRIFICED ON DAY 3 OF 336. SACRIFICED ON DAY 2 OF 312 . 297 . 272 . 262 . 309 . 299. SACRIFICED ON 291. SACRIFICED ON DAY 1 OF 282 . SACRIFICED ON DAY 3 OF 278 . 275 . 287. SACRIFICED ON DAY 3 OF 278 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS DAY 5 OF LACTATION DUE TO NO SURVIVING LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS DAY 5 OF LACTATION DUE TO NO SURVIVING LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS PUPS PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-180 000254 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 8): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 8 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 266. 334 . 295 . 300 . 306 . 325. 312 . 297. 337. 318. 316. 343 . 282 . 328. 304 . 333 . 306 . 286. 311. 321. 306. 333 . 304 . 325. 333 . 322 . 325. 334 . 340. 325. 344 . 383 . 281. 333 . 297 . 337. 312. 312. 345 . 343 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 0.4 MG/KG PFOS 418-018:PAGE B-18 000255 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 28 (PAGE 9): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 9 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 259 . 208 . 316 . 346. 325. 335 . 269 . 283 . 277 . 290. NOT PREGNANT 298 . 298 . 283 . 279 . NOT PREGNANT 325 . 324 . NOT PREGNANT 341. 356. 288 . 296 . 340. 364 . 287. 326 . 327. 355 . 296 . 278. 313 . 326 . 322 . 332 . 302 . 245. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 0.8 MG/KG PFOS 9SZ000 418-018 :PAGE B-182 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 8 (PAGE 10): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 10 1 MG/KG PFOS 110X9 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 307 . 306 . 322 . 336. 287 . 282 . NOT PREGNANT 294 . 281. 327 . 337 . 285 . 314 . 302 . 308 . 276. 277 . 310. 300 . 285 . 275 . 315 . 317. 309. 322 . 308 . 305. 307. 323 . 322 . 323 . 301. 310. 284 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 317. 324 . 325. 331. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-183 000257 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 11) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP l: 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 295 . 304 . NOT PREGNANT 306 . 302 . 325 . 321. 282. 273 . 299. 296. 282. 274 . 307 . 338. 270 . 280. 307. 306 . 270 . 286. NOT PREGNANT 282 . 287 . 320. 347. 308 . 329. 331. 333 . 308 . 314 . 295 , 310 . 261. 276 . 318 . 325. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 1.2 MG/KG PFOS 418-018:PAGE B-184 000258 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 12) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11041 11042 11043 11048 11049 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 NOT PREGNANT 297 . 292 . 295 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS NOT PREGNANT 277 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 296 . 304 . 278 . 293 . 304 . 316. 297. 290 . 294 . 281. 263 . 288 . 332 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 303 , 316. 310. 300 . 274 . 279. 314 . 305. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 302 . 289. 343 , 352 . NOT PREGNANT 290. 276. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-185 000259 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 13) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # 11055 11057 11059 11061 11064 11065 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 DAY 1 5 DOSAGE GROUP 13 ! MG/KG PFOS 301. SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 267 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 275 . 286. 313 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 334 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 285 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 271. 272 . 282 . 293 . 278 . 279 . 313 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 259 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 275, SACRIFICED ON DAY i OF LACTATION DUE TO NO SURVIVING PUPS 298 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 274 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 317. 284 . 313 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 268 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 301. 289. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS NOT PREGNANT 306 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 418-018:PAGE B-186 000260 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 1): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT It DOSAGE GROUP 1 VEHICLE CONTROL 10901 10902 10903 10904 10905 10906 10907 10908 10909 10910 10911 10912 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 141. 122 . 137 . c 116 . 141. 137. 117 . 14 8 . 163 . 144 . c 211. 130. 100 . 142 . 137. 157. 137 . 136. 126. 146. 123 . 143 . 108 . 156 . 116 . 132 . 128 . 120. 130. 142 . 117 . 138 . 129. 118 . 136. 146 . 134 . 138. 117 . 123. 105 . 153 . 127. 149 . 135. 142. 128 . 146. 125 . 147. 123 . 161. 121. 143 . 123 . 125 . 131. 144 . 130 . 136. 127 . 117. 133 . 150. 128 . 128 . 127 . 115 . 95 . 155 . 130 . 150. 129. 140 . 125. 145 . 133 . 156 . 123 . 170. 101. 148 . 117 . 123 . 135 . 137. 136. 135 . 130. 122 . 131. 149. 116 . 141. 121. 119. 117 . 153 . 120 . 168. 139. 134 . 126 . 130 . 124 . 143. 115 . 158 . 117 . 141. 123 . 108. 129 . 143 . 129 . 140. 127. 107. 131. 147 . 123 . 155. 119. 122 . 115 . 141. 136 . 176 . 135 . 135. 129 . 140. 130. 137 . 105 . 147 . 124 . 134 . 102 . 100. 107. 109. 96 . 109. 102 . 104 . 100. 127. 105 . 121. 94 . 93 . 103 . 126 . 97 . 129 . 124 . 117 . 103 . Ill. 119. 129. 97. 139. 92. Ill. ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-187 000261 PROTOCOL 418-018; ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 2): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8 - 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL 1091S 10916 10917 10918 10919 10920 10921 10922 10923 10924 10925 10926 10927 10928 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 151. c 147 . 141. 140. c 152 . 149. 139. 146. 142 . 146. 154 . 127. 116 . 155 . 130 . 140 . 137 . 141. 146. 121. 126. 135. 138 . 127. 130. 141. 135. 179. 131 . 136. 125 . 138 . 139. 137 . 124 . 131. 138 . 159. 137 . 124 . 115 . 184 . 124 . 162 . 145 . 152 . 148. 136. 134 . 141. 153 . 135 . 142 . 146 . 123 . c 149. 130 . 120 . 158 . 144 . 136 . 145. 130. 129 . 147 . 134 . 124 . 119. 184 . 119. 157. 160. 134 . 146 . 137 . 144 . 142. 168 . 153 . 128 . 141. 123 . 157 . 127 . 145 . 136. 135. 141. 135. 159. 139. 131. 145 . 141. 128 . 123 . 158. 118 . 157. 166 . 132 . 143 . 133 . 123 . 135. 151. 143 . 128 . 163 . 120. 154 . 129. 130 . 128 . 143 . 143 . 131. 148. 132. 117. 141. 137 . 124 . 115 . c 123 . 166 . 168 . 134 . 138 . 140. 118 . 133 . c 135 . 123 . 174 . 104 . 98. 99. 106 . 97 . 112 . 107. 119. 119. 94 . 104 . 122 . Ill. 105. 99. 129. 102 . 136 . 148 . 120 . 124 . 110. 103 . 113 . 124 . 110. 108 . 127. ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-188 000262 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 2 9 (PAGE 3) : FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE: GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 10930 10931 10932 10933 10934 10935 10936 10937 10938 10939 10940 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 122 . 113 , 119 . 121. 131. 132 . 137 . 134 . 130 . 115 . 110 . Ill . 138 . 121. 122 . 124 . 119. 127 . 121. 118. 149 . 130. 124 . 122 . 120. 99. 105 . 106 . FOUND DEAD ON DAY 2 OF STUDY 139. 130 . 116. 121. 143. 129. 130 . 124 . 148 . 125. 118. Ill . 132 . 135 . 138. c 133 . 121. 123 . 111. 134 . 128 . 131. 120 . 120 . 121. 120. 119 . 144 . 150. 153 . 154 . 154 . 139. c 116 . 130. 132 . 143. 135 . 149. 147. 136. 142. 146. 151. 135 . 155 . 143 . 115. 130 . 134 . 141. 151. 154 . 154 . 126 . 141. 138. 125. 136 . 138 . 135. 107. 94 . 90. 119. 92 . 129 . 142 . 135 . 138 . 129 . 122 . 115 . 134 . 123 . 223 . 180 . 125 . 162 . 165. 170. 160 . 117 . 129 . 104 . 128 . 112 . 117. 100. 122 . 135 . 113 . 148 . 115. 123. 112 . 134 . 149 . 146 . 145 . 155 . 129 . 147 . 129. 119. 115. 140. 133 . 136 . 156 . 102 . 103 . 89. 91. 103 . 97. 82 . 99. 102 . 87. c 92 . 98. 95 . 126. 102 . 115. 108 . 121. 110 . 123 . 105 . 108 . 84 . 117. 109. 121. 140 . ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-189 000263 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 9 (PAGE 4): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 10944 10945 10946 10947 10948 10949 10950 10951 10952 10953 10954 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 11556 11557 129. 154 . 144 . 153 . 130 . 130 . 152 . 135 . 141. 128 . 126 . 123 . 126 . 132 . 133 . 149. 136. 127 . 126. 131. 128 . 141. 150 . d 133 . 145 . 136 . 144 . 124 . 123 . 136 . 139. 126 . 128 . 138 . 131. 161. 128 . 123 . 124 . 122 . 121. 133 . 154 . 90. 131. 131. 125. 116 . 137. 149 . 134 . 137 . 163 . 121 . 146 . 129. 110. 136 . 129 . 118 . 124 . 142 . 123 . c 132 . 117 . 124 . 114 . 130. 140 . 155 . 136 . 125 . 165. 142 . 148. 139. 142 . 136 . 132 . 155. 118 . 142 . 120. 114 . 130. 136 . 113 . 128 . 131. 123 . 127 . 125. 115 . 123 . 110 . 134 . 129 . 156 . 131. 126 . 124 . 133 . c 130 . 146 . 115 . 127 . c 155 . 143 . 118 . 109 . 124 . 174 . 107 . 120 . 126 . 119. 119 . 125 . 106 . 124 . 112 . 118. 127. C 120 . 141 . 120. 123 . 177 . 136 . 144 . 135 . 123 . c 153 . 134 . 95 . 89 . 99. 104 . 93 . 96. 94 . 95 . 100 . 99 . 96 . 102 . 93 . 94 . 103 . 155 . 107 . 102 . 102 . 106. 112 . 115 . 101 . 108 . 106 . 142 . 108 . 106. AL L WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a . Last value recorded before cohabitation, b. First value recorded after cohabitation. c . Spilled feed precluded the calculation of this value. d. Value was not recorded. *92000 418-018 :PAGE B-190 418-018:PAGE B000265 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 5): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 122. 134 . 131. 135 . 114 . 133 . 135 . 143 . 124. 147. 125 . 130. 129. 150 . 125 . 143 . 134 . 137. 152 . 149. 128 . 129. 130. 153 . 110. 133 . 127 . 143 . 120 . 140 . 125 . 125 . 131. 138 . 128 . 139 . 126 . 142 . 125. 125 . 127 . 148 . 129. c 146. 145 . 144 . 169. 131. 117 . 133 . 154 . 109. 146 . 129. 140 . 122 . 121. 135 . 94 . 142 . 144 . 157 . 129. 129. 124 . 127 . 113 . 150 . 146 . 150. 142 . 140 . 149. 163 . 116 . 143 . 158 . 143 . 127 . 133 . 129 . 151. 125. 141. 136 . 160 . 127 . 128 . 122 . 122 . 109 . 174 . 152. 173 . 137. 129. 136 . 136 . 114 . 135 . 136. 134 . 113 . 122 . 126 . 127. 104 . 159. 155 . 163 . 124 . 126 . 131. 133 . 105. 151. 144 . c 121. 136 . 149 . 152 . 121. 163 . 159. 147 . 115 . MORIBUND SACRIFICED ON DAY 16 OF STUDY 166 . 165. 173 . 138 . 130 . 126 . 134 . 115 . 117 . 126. 123 . 106 . 146 . 151. c 133 . 141 . 128 . 176 . 124 . 141 . 105 . 113 . 94 . 137 . 142 . 155. 120 . 126 . 133 . 158. 101. 147 . 143 . 161. 119. ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. vo PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 6): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 10972 10973 10974 10975 10976 10977 10978 10979 10980 10981 10982 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 120 . 144 . 124 . 124 . 130 . 155 . 132 . 126. 125 . 133 . 126 . 122. 153 . 117 . 103 . 131. 127 . 129. 141 . 142 . 140 . 136 . 119 . 145 . 134 . 131. 133 . 126 . 117 . 150 . 129 . 141. 136 . 137 . 123 . 122 . 130. 123 . 121. 123 . 134 . 127. 104 . 129 . 136. 137. 132 . 129. 138. 147 . 132 . 133 . 128 . 126 . 133 . 142 . 113 . 135 . 127 . 127 . 134 . 136. 117. 123 . 165 . 127. 121. 131. 114 . 126 . 105. 131. 148 . 157. 164 . 128. 124 . 139. 126 . 150 . 135. 120 . 133 . 127. 105. 135 . 122 . 120. 127. 121. 116 . 122 . 140 . 113 . 116. 115. 120 . 127. 106 . 132 . 121. 133 . 133 . 131. 130 . 136 . 130 . 154 . 127. 127 . 136 . 122 . 106 . 142 . 127 . 134 . 126 . 126 . 118 . 123 . C 124 . 112 . 119 . 125 . 130 . c 144 . 129. 137 . 130 . 128 . 134 . 149. 128 . c 131. c 132 . c 88 . 114 . 103 . 103 . 107 . 93 . 89 . 103 . Ill . 118 . 90 . 98 . 96. 107 . 90 . 115. 97 . 112 . 115. 109. 113 . 115. 102 . 97 . 107 . 104 . 108 . 100 . ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-192 000266 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 29 (PAGE 7): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8 - 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT tt DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10985 10986 10987 10988 10989 10990 10991 10992 10993 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 135 . 154 . 140. 149 . 120 . 118 . 123 , 123 . 137. 142 . 129 . 131. 109 . 139. 125 . 142 . 138. 146. 138. 124 . 133 . 119 . 133 . 150 . 123 . 138 . 130. 132 . 130. 145. 139. 146 . 118 . 118. 121. 118. 137 . 132 . 129. 133 . 113 . 149. 125 . 143 . 147. 156. 162 . 128 . 133 . 110 . 133 . 148 . 123 . 134 . 117 . 118 . C 134 . 137 . 136 . 117. 122 . 123 . 118 . 146. 126 . 119 . 118 . 109 . 142 . 147 . 129. 131. 161. 164 . 131. 130. 115. 140 . 141. 162 . 140. 128 . 118 . 132. 127 . 126 . 125 . Ill. 129. 112 . 112 . 131. 116. 109 . 113 . 106 . 138 . 131. 140 . 135 . 134 . 156 . 124 . 143 . 98 . 130. 129 . 117 . 126 . 108 . 110. 133 . 130 . 130 . c 116 . 118. 122 . 116 . 134 . 129. 112 . 118 . 106 . 143 . 136 . 159 . 125. 178 . c 133 . 130. 101. 131 . 129. c 126. 137 . 102 . 112 . 99 . 103 . 99. 89. 94 . 102 . 85 . 116. 96 . 100 . 95 . 94 . 121. 108 . Ill . 96. 113 . 108 . 114 . 107 . 96. 106. Ill. 114 . 106 . 125 . 92 . ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 9Z000 418-018:PAGE B-193 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 8) : FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 8 0.4 MG/KG PFOS 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 121. 138 . 122 . 127 . 132 . 143 . 165 . 156. 158 . c 147 . 148 . 119. 138. 132 . 150. 138. 125 . 126. 129. 121. 135. 127 . 131. 134 . 148. 169 . 141 . 137. 153 . 159. 149. 108 . 43. 140 . 146. 127. 122 . 132 . 135. 117. 131. 124 . 125 . 142 . 140 . 135 . c 159. 157. 13 9. 122 . 122. 147. 143 . 147. 133 . 130 . 125. 150. 122 . 129 . 116. 124 . 150 . 142 . 135. 142 . 165. 152 . 150. 134 . 116. 122. 138. 153 . 128 . 123 . 127. 150. 124 . 130 . 116 . 131 . 151. 155 . c 167. 156. 153 . 171. 148 . 140 . 136. 146 . c 130 . 123. 135 . c 94 . 109 . 107 . 98 . 117 . 128 . 136 . 131. 142 . 116. 123 . 123 . 100. 109. 121. 120. 115. 101. 116 . 120. ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-194 000268 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 2 9 (PAGE 9) : FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT tt DOSAGE GROUP 9 0.8 MG/KG PFOS 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 132. c 139 . 120 . 121. 137. 141. 117. 157 . 140. 142 . 149. 113 . 151. 124 . 124 . 122 . 131 . 128 . 153 . 112 . 151. 135 . 123 . 122 . 133 . 137. 112 . 150 . 139. 135. 142 . 122 . c 138 . 129 . 132. 138. 131. 136. 113 . 135. 139 . 117 . 127 . 138 . 141. 122 . c 141 . 143 . 157 . 128 . c 138 . 144 . 121. 145 . 135. 131. 116 . 158. 141. 116. 124 . 133 . 140 . 118 . c 153 . 125 . 146. 115 . 160 . 136 . 144 . 101. 148 . 148 . 107. 113 . c 131. 120. 117 . 133. 131. 112. c 140. 125. 146. 115. 154 . 129 . 124 . 106 . 131. 161. 125 . 94 . 96 . 112 . 92. 104 . 114. 119 . 87 . 159. 103 . 106. 116. 99. 118. 112 . 105 . 101. 117. 124 . 107 . 146. 97. 103. 69. AL L WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c Spilled feed precluded the calculation of this value. 418-018:PAGE B-195 000269 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 10): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT 0 DOSAGE GROUP 10 1 MG/KG PFOS 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 122 . 147 . 172 . 120. 126. 148 . 144 . 113 . 110. 136. 121. 140. 127 . 127 . 125 . 145 . 117 . 149 . 127. 133 . 115 . 148 . c 120 . 130 . 148 . 121. 128 . 115 . 134 . 125 . 140 . 128 . 206 . 122 . 136 . 134 . 150 . 138. 144 . 125 . 146 . 171 . 135 . 139 . 186. 136 . 137. 121 . 140. 130. 129 . 141. 165 . 137. 150 . 137. 153 . 137. 146 . 124 . 143 . 161. 139. 137 . 160 . 139 . 138 . 120 . 140 . 149. 124 . 126 . 145. 130. 133 . 128 . 165 . 142 . 130. 118 . 151 . 168 . 132 . 126. c 135 . 142 . 119. 143 . 121. 127 . 170. 130 . 133 . 145 . 126 . 160 . 136. 139 . 90 . c 117 . 100 . 102 . 135 . 114 . 91. 95 . Ill . 104 . 101. 121. 102 . 121. 120 . 111. 122 . 112 . 121. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018-.PAGE B-196 000270 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 11): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 140 . 147 . 133 . 137 . 133 . 123 . 130. 155. 121. 134 . 147 . 140 . 125 . 136 . 140. 142 . 128 . 125 . 109 . 142 . 128 . 137 . 138 . 136. 136 . 114 . 120 . 142 . 116. 132 . 118. 160. 132 . 146. 133 . 144 . 125 . 126 . 106. 132 . 125. 143 . 143 . 143 . c 125. 120. c 128 . 131. 128 . c 132 . 155 . 142 . 145 . 128 . 132 . 119. 136 . 148 . 131. 147. 141. 140 . 124 . 117 . 124 . 136 . 126 . 118. c 125. 141. 145 . 152 . 133 . 116 . 111. 125 . 133 . 126 . 144 . 131. 139. 117 . 124 . 115 . 141. 123 . 118 . c 119. 148. 132 . 145 . 130 . 108 . 108 . 130 . 83 . 108. 101. 121. 118. 93 . 90. 104 . 101. 103 . 101. c 91. 118 . 121. 127 . 115 . 94 . 87 . 103 . 132. 101. 133. 64. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-197 000271 PROTOCOL 418-0X8: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS {SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 29 (PAGE 12): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11040 11041 11042 11043 11044 11045 11046 11047 11048 11049 11050 11051 11052 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 149 . 130. 144 . 143 . 126 . 138. 147 . 136 . 122 . 116 . 125 . 143 . 133 . 137. 155. 156 . 142 . 148 . 128 . 173 . 137 . 144 . 103 . 149 . 129. 149. 148 . 158 . 143 . 131 . 134 . 137 . 126 . 136. 131. 130 . 116 . 115 . 127 . 130 . 122 . 131. 127. 160 . 138 . 136. 126 . 154 . 131 . 141 . 98 . 154 . 129 . 150 . 146 . 112 . 120 . 137 . 140. 141. 127 . 139. 139. c 116 . 120 . 138 . 128 . 122 . 138. 124 . 163 . 153 . 140 . 117 . 160 . 128 . 135 . 104 . 162 . 119. 141. 141 . 121. 135 . 137 . 132 . 130 . 131. 125 . 143 . c 129 . 125. 132 . 134 . 120. 135 . 133 . 146 . 128 . 123 . 114 . 14 6 . 116 . 130 . 95 . 147 . 126 . 133 . 142 . 115 . 132 . 120 . 132 . 127 . 135 . 128 . 119. 131. 116 . 117 . 126 . 116 . 109. 121. 127 . 148 . 121. 132 . 133 . 179 . 118 . 140 . 108 . 133 . 105 . 149 . 151. 116 . 101 101 101 100 94 105 100 93 97 91 101 95 99 93 102 116 100 97 91 128 113 115 90 123 91 113 108 93 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-198 000272 000273 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 29 (PAGE 13): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 13 2 MG/KG PFOS 11053 11054 11055 11056 11057 11058 11059 11060 11061 11062 11063 11064 11065 11066 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 122 . 142. 122 . 126 . 120. 139. 138 . 120. 132 . 139. 127. 136 . 103 . 155 . 127 . 150 . c 143 . 129. 128 . 135 . 166 . 133 . 148 . 122 . 124 . 136 . 136. 114 . 134 . 115 . 107 . 113 . 131. in. 119. 128 . 122 . 126. 137 . 109. 132 . 121. 134 . 143 . 139. 136. 116. 130 . 133 . 135 . 143 . 126 . 124 . 124 . 135. 126. 142. 112 . 101. 115. 139 . 127 . 124 . 135 . 128. 125 . c 115 . 127 . 121. 116 . 149. 136. 119 . 132 . 125. 129 . 140. 152 . 119 . 130 . 130. 134 . 121. 143 . 112 . 103 . 116. 115 . 151. 133 . 131. 124 . 118 . 140 . 116 . 121. 121. 107 . 126 . 130 . 122 . 125. 114 . 118 . 131. 141. 106 . 113 . 126 . 132 . 112 . c 111. 105 . 109. 110. 122. 113 . 127 . 124 . 114 . c 106 . 121. in. 119. 135 . 137 . 117 . 123 . Ill . 126 . 127. 144 . 113 . 108. 129. 131. 90 . 117 . 92 . 73 . 93 . 98 . 94 . 82 . 101. C 92 . 99 . 100 . 98 . 100 . 95 . 95. 112 . 101. 94 . 96 . 102 . 113 . 117 . 88 . 91. 100. 105 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. ~S HOVd:8IO'8li7 iO PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 30 (PAGE 1) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 1 VEHICLE CONTROL 10901 P 10902 P 10903 P 10904 P 10905 P 10906 P 10907 P 10908 P 10909 P 10910 P 10911 P 10912 NP 10913 P 10914 P 11501 P 11502 P 11503 P 11504 P 11505 P 11506 P 11507 P 11508 P 11509 P 11510 P 11511 P 11512 P 11513 P 11514 P 144 . 140 . 157 . 164 . 170 . 159. 162. 147 . 169 . 186 . 167. 173 . 131. 141. 143 . 170. 176 . 162 . 162 . 172 . 153 . 145 . 165 . 149 . 132 . 169. 123 . 164 . 157. 147. 162 . 178 . 171. 159 . 169. 161. 147 . 186 . 153 . 196 . 137. 139. 135 . 169. 169. 176. 166. 165 . 169. 162 . 174 . 164 . 137 . 185 . 131. 163 . 156 . 174 . 189 . 183 . 201. 175 . 179. 171 . 150 . 193. 164 . 150. 156 . 154 . 154 . 170 . 144 . 167 . 153 . 186 . 192 . 172 . 185 . 179. 149 . 183 . 155 . 163 . a P = PREGNANT NP * NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value was not recorded. 418-018:PAGE B-200 000274 9 Z * * 0 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 2) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7-14 14 - 21 21 - 25 RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10915 P 10916 P 10917 P 10918 P 10919 P 10920 P 10921 P 10922 P 10923 P 10924 P 10925 P 10926 P 10927 P 10928 P 11515 P 11516 P 11517 P 11518 P 11519 P 11520 P 11521 P 11522 NP 11523 p 11524 P 11525 P 11526 p 11527 P 11528 P 131. 147 . 136 . 167 . 161. 144 . 152 . 157 . 152 . 163 . 148 . 158. 146 . 151. 130. 166. 148 . 156 . 167 . 150 . 172 . 139. 135 . 156. 172 . 142 . 123 . 160. 155 . 154 . 168 . 196 . 155 . 166. 187 . 195 . 183 . 163 . 186. 206. 162 . 189 . 164 . 152 . 159. 154 . 173. 175 . 165 . 188 . MORIBUND SACRIFICED ON DAY 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 154 . 163 . 160. 185. 132 . 138 . 157 . 173 . 150. 152 . 14 8 . 184 . 184 . 180 . 155 . 155. 159. 166 . 134 . 106. 57. 138 . 154 . a 162 . 188. 191. a 153 . 155 . 176 . 166 . 169. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. 418-018:PAGE B-2 O PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 30 (PAGE 3) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT tt DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 P 10930 P 10931 P 10932 P 10933 P 10934 P 10935 P 10936 10937 P 10938 P 10939 NP 10940 P 10941 P 10942 P 11529 P 11530 P 11531 NP 11532 P 11533 P 11534 P 11535 P 11536 P 11537 P 11538 P 11539 P 11540 P 11541 P 11542 P 11543 P 127. 135. 172 . 126 . 142 . 173 . 119. 131. 168 . 121. 158 . 191. 116 . 133 . 190. 139 . 151. 179. 105 . 124 . 152. FOUND DEAD ON DAY 2 OF STUDY 113 . 138. 162 . 101. 156. 130. 118 . a a 167. 160. 230 . 115. 139. 114 . 155. 114 . 136. 161. 139. 159. 173 . 109 . 120. b 187. 139. 153 . 183 . 125 . 137. 165. 133 . 171. 174 . 124 . 146 . 174 . 141 . 169. 183 . 123 . 146 . 179 . 119. 140 . 165. 118 . 122 . 144 . 141. 152 . 182 . 141. 143 . 167 . 127. 148. 166 . 168 . 187 . P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. value was not recorded. b. Spilled feed precluded the calculation of this value. $AZ*O 418-018:PAGE B-202 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC AC ID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 4): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT a DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 P 10944 P 10945 P 10946 P 10947 P 10948 P 10949 P 10950 P 10951 P 10952 P 10953 P 10954 P 10955 P 10956 P 11544 P 11545 P 11546 P 11547 P 11548 NP 11549 p 11550 p 11551 P 11552 P 11553 P 11554 P 11555 p 11556 P 11557 P 117 . 101 . 127 . 194 . 123 . 131. 142 . 135. 118 . 147. 104 . 127 . 117 . 142 . 119 . 175 . 130. 147 . 156 . 134 . 119 . 140 . 136 . 133 . 131. 145 . 144 . 136 . 139. 131 . 165 . a 139. 117 . 154 . 174 . 155 . 119 . 138 . 140 . 153 . 162 . 142 . 171. 140 . 156. 116. 140 . 132 . 149. 146 . 146 . 137 . a 143 . 151. 165. 185 . 192 . 168 . 176. 84 . 184 . 188 . 138 . 178 . 181. 166 . 189. 161. 164 . 97. 164 . 156 . 183 . 167. 149 . 142 . 215 . 168 . 138 . 61. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. 418-018:PAGE B-203 000277 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 5): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 P 10958 P 10959 P 10960 P 10961 P 10962 P 10963 P 10964 P 10965 P 10966 P 10967 P 10968 P 10969 P 10970 P 11558 P 11559 P 11560 NP 11561 p 11562 11563 P 11564 p 11565 P 11566 NP 11567 P 11568 p 11569 p 11570 P 11571 p 119 . 152 . 160 . 185 . 174 . 153 . 183 . 161. 144 . a 170 . 160 . 152 . 164 . 126 . 191. 140 . 191. MORIBUND a 159 . 155 . 178 . 176. 135 , 161 . 132 . 186 . 153 . 168 . 159. 184 . 165 . 198 . 210 . 215 . 171. 191. 153 . 172 . 178 . 207. 183 . 203 . 142 . 167 . a 174 . 165. 153 . 166. 184 . 193 . 178 . 142 . 161. 196 . 176 . 127 . 149. 177 . 162 . SACRIFICED ON DAY 203 . 192 . 156 . 156 . 156 . 168 . 153 . 114. 182 . 207 . 148 . 152 . 167 . 167 . 146. 146 . 183 . 203 . 69. 16 OF 74. STUDY P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. 418-018:PAGE B-204 00027 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 6) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 P 10972 P 10973 P 10974 P 10975 P 10976 P 10977 P 10978 P 10979 P 10980 P 10981 P 10982 P 10983 P 10984 P 11572 P 11573 P 11574 P 11575 P 11576 P 11577 P 11578 P 11579 P 11580 P 11581 P 11582 P 11583 P 11584 P 11585 P 127 . 151. 122 . 137. 132 . 120 . 117 . 125 . 155. 118 . 94 . 120 . 133 . 147. 116. 146 . 126. 141. 127 . 146. 147. 149 . 137 . 163 . 146 . 125. 135. 113 . 122 . 170. 147. 157 . 133 . 128 . 128 . 134 . 170. 130 . 139. 138 . 146. 168 . 139. 156. 137. 168 . 166 . 158. 150 . 153 . 171. 144 . 151. 156. a 141 . 149 198 160 186 174 158 139 181 179 164 166 199 151 175 176 181 175 180 178 160 191 188 172 173 160 164 P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. 418-018:PAGE B-205 000279 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 7) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7-14 1 4 - 2 1 21 - 25 RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10985 P 10986 P 10987 P 10988 P 10989 P 10990 P 10991 P 10992 P 10993 P 10994 P 10995 P 10996 P 10997 P 10998 NP 11586 p 11587 P 11588 P 11589 p 11590 P 11591 P 11592 P 11593 P 11594 p 11595 p 11596 P 11597 P 11598 P 11599 P 134 . 114 . 131. 118 . 116 . 124 . 117. 110 . 116 . 120 . 127 . 122 . 112 . 135. 131. 146. 118 . 156 . 142 . 143 . 137 . 118 . 125 . 148 . 154 . 138 . 133 . 108 . 156 . 136 . 140 . 138. 129. 146 . 136 . 135 . 161. 135 . 152 . 131. 129 . 123 . 152 . 169. 141. a 158 . 172 . 138 . 139 . 150 . 172 . 150. 155 . 138 . 139. 182 . 188 . 197 . 154 . 161. 159. 154 . 168 . 188 . 202 . 181. 167. 155. 127. 158 . 195 . 202 . 158 . 194 . 179 . 154 . 185 . 184 . 177 . 177 . 165 . 164 . 86. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. 418-018:PAGE B-206 000280 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 8): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 8 0.4 MG/KG PFOS 10999 P 11000 P 11001 P 11002 P 11003 P 11004 P 11005 P 11006 P 11007 P 11008 P 11600 P 11601 P 11602 P 11603 P 11604 P 11605 P 11606 P 11607 P 11608 P 11609 P 142 . 160. 132 . 165 . 160. 159. 167. 158 . 205. 169 . 176. 169. 148. 159. 144 . 164. 14 9. 133 . 166 . 161. 170 . 168 . 147 . 163 . 155 . 180. 165 . 177. a 153 . 170. 169. 148 . 161. 151. 169. 153 . 147. 155. 180. 141 174 174 188 212 166 140 171 199 179 169 173 164 183 172 150 168 162 161 193 P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . a. Spilled feed precluded the calculation of this value. TffZOOO 418-018:PAGE B-207 PROTOCOL 418-018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 9) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 9 0.8 MG/KG PFOS 11009 P 11010 P 11011 p 11012 P 11013 p 11014 NP 11015 P 11016 P 11017 NP 11018 p 11610 NP 11611 p 11612 p 11613 P 11614 P 11615 p 11616 p 11617 P 11618 P 11619 P 130 . 132 . 148 . 162 . 191 . 133 . 168 . 188 . 132 . 137 . 165 . 141. 155 . MATING NOT CONFIRMED 138 . 164 . 187 . 128. 152. 155 . 166 . 98 . 158 . 165 . 182 . 129 . 148 . 139 . 171. 183 . 179. 137 . 141 . 168 . 181. 190 . 210 . 146 . 156 . 181 . 135 . 155 . 157 . 122 . 148 . 173 . 142 . 163 . a 164 . 164 . 172 . 137 . 142 . 177 . 66 . 69. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. 418-018:PAGE B-208 000282 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER : T-6295.25) TABLE 30 (PAGE 10): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT tf DOSAGE GROUP 10 1 MG/KG PFOS 11019 P 11020 P 11021 P 11022 NP 11023 P 11024 P 11025 P 11026 P 11027 P 11028 P 11620 P 11621 P 11622 P 11623 P 11624 P 11625 P 11626 P 11627 P 11628 P 11629 P 136 . 147. 162 . 155 . 140 , 169 . 151 . 140 . 124 . 133 . 140 , 170. 159 . 135 . 150. 164 . 129. 14 5 . 127 . 136 . 122 . 152 . 163 . 132 . 153 . 183 . 143 . 161. 162 . 144 . 95. 147. 169. 152 . 168. 167 . 147 . 156. 159. 150 . 171. 173. 195 . 107. 179 . 207 . 170. 166. 177. 190. 169. 170. 186 . 167. 188 . 185 . 160. 119. 182 . 173 . 72. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 418-018:PAGE B-209 000283 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 30 (PAGE 11): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7-14 14-21 21-25 RAT # DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 P 11030 NP 11031 P 11032 P 11033 P 11034 P 11035 P 11036 P 11037 p 11038 P 11630 P 11631 NP 11632 P 11633 p 11634 P 11635 p 11636 p 11637 P 11638 P 11639 p 135 . 185 . 204 . MATING NOT CONFIRMED 144 . 157 . 169. 137 . 164 . 195 . 134 . 174 . 197 . 124 . 134 . 162 . 128 . 149 . 166 . 140 . 151. 177 . 117 . 146 . 166 . 139. 144 . 177. 137 . 142 . 147. 153 . a 8 .b 114 . 141. 150. 161. 165 . 195 . 153 . 164 . 153 . 161. 167. 187. 151. 169. 193 . 128 . 134 . 147. 136. 140. 164 . 128 . 154 . 164 . c P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . a. Spilled feed precluded the calculation of this value. b. Value appeared incorrectly recorded and was excluded from group averages and statistical analyses. c. Value appeared incorrectly recorded and could not be calculated. 418-018:PAGE B-210 000284 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 12): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7-14 1 4 - 2 1 21 -- 25 RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 NP 11040 P 11041 P 11042 P 11043 NP 11044 P 11045 P 11046 P 11047 P 11048 P 11049 P 11050 P 11051 P 11052 P 11640 P 11641 P 11642 P 11643 P 11644 P 11645 P 11646 P 11647 P 11648 P 11649 P 11650 P 11651 P 11652 NP 11653 P MATING NOT CONFIRMED 103 . 126. 188 137 . 152 . 122 . 149 . 95 . 106 . 103 119. 147. 195 139. 152 . 180 129. 152 . 173 124 . 129. 181 133 . 160 . 152 141. 150. 186 121. 134 . 192 117. 142 . 154 141. 153 . 202 128 . 143 . 169 153 . 168 . 166 122 . 131. 167 131. 145 . 173 124 . 136. 165 171. 208 . 211 141. 151. 186 141 . 176 . 182 113 . 126 . 150 152 . 158 . 182 133 . 120. 171 152 . 175 . 187 181. 132 . 116 133 . 137 . 157 P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 418-018:PAGE B-2 000285 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 13): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 13 2 MG/KG PFOS 11053 P 11054 P 11055 P 11056 P 11057 P 11058 P 11059 P 11060 P 11061 P 11062 P 11063 P 11064 P 11065 P 11066 P 11654 P 11655 P 11656 P 11657 P 11658 P 11659 P 11660 P 11661 P 11662 P 11663 P 11664 P 11665 P 11666 NP 11667 P 106 . 127 . 130 . 114 . 127 . 123 . 115 . 115. 133 . 125 . 122 . 171. 114 . 134 . 128 . 158 . b 140. 131. 120 . 119. 133 . 148 . 153 . 126. 123 . 149. 149. 128 . 154 . 152 . 125 . 139 . 130 . 135. 135 . 162 . 149 . 125 . 271. 145. 170 . 144 . 14 5 . 153 . 156 . 137 . 123 . 133 . 146 . 140 . 163 . 124 . 150. 106. 146 . 172 191 195 154 186 189 155 195 198 a 177 134 159 206 166 171 159 180 139 141 175 161 164 184 147 171 99 166 P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value was not recorded. b. Spilled feed precluded the calculation of this value. 418-018:PAGE B-212 000286 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 1 VEHICLE CONTROL 10906 10909 10911 10913 10914 11501 11502 11503 11504 USOS 11506 11507 11508 11509 11510 11511 11512 11513 11514 121. 113 . 116 . a 119. 86 . 104 . 98 . 150 . 14 8. 142 . 114 . 94 . 118 . 137. 107 . 99. 93 . 116. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION a. Spilled feed precluded the calculation of this value. 418-018 :PAGE B-213 000287 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 2) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10921 10922 10923 10924 10926 10927 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 128 . 138 . 143 . 106 . MORIBUND SACRIFICED ON DAY 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 122 . 108 . 130 . 145 . 86. 74 . 118 . 87. NOT PREGNANT 130 . 112 . 86 . 96 . 121. 117 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-214 000288 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 3) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 3 3 .6 MG/KG PFOS! + MEVALONIC ACID 10929 10936 10938 10939 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 79. . FOUND DEAD ON DAY 2 OF STUDY 58 . NOT PREGNANT SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 107 . 88 . 90 . NOT PREGNANT SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 65 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS a 104 . 76. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 49 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 56 . 115 . ALL WEIGHTS WERE RECORDED IN GRAMS (G) DAYS = DAYS OF LACTATION a. Spilled feed precluded the calculation of this v a l u e . 418-018:PAGE B-215 000289 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 31 (PAGE 4) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 4 ! MG/KG PFOS i- MEVALONIC ACID 10943 10944 10948 10952 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 11556 11557 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 47. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY i OF LACTATION DUE TO NO SURVIVING PUPS 55 . NOT PREGNANT SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 65. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY i OF LACTATION DUE TO NO SURVIVING PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-216 000290 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 5): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 121. 190 . 127. Ill. 123 . 122 . 112 . 139. NOT PREGNANT 140. MORIBUND SACRIFICED ON DAY 16 OF STUDY 139. 99. 129. NOT PREGNANT 124 . 118. 124 . 97 . 131. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-217 0002SJL PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 6) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 10973 10977 10979 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 66 . 105 . 99. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 57 . 119 . 87. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 89. 51. 70 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 58 . 83 . 109. 42. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-218 000292 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 7): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT tf DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10986 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 SACRIFICED ON DAY 3 OF SACRIFICED ON DAY 3 OF 56 . SACRIFICED ON DAY 2 OF 83 . NOT PREGNANT SACRIFICED ON DAY 3 OF SACRIFICED ON DAY 3 OF SACRIFICED ON DAY 2 OF 53. SACRIFICED ON SACRIFICED ON DAY 3 OF SACRIFICED ON DAY 2 OF 67 . 39. 50. SACRIFICED ON SACRIFICED ON DAY 1 OF SACRIFICED ON DAY 3 OF 61. SACRIFICED ON DAY 3 OF SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS DAY 5 OF LACTATION DUE TO NO SURVIVING LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS DAY 5 OF LACTATION DUE TO NO SURVIVING LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS LACTATION DUE TO NO SURVIVING PUPS PUPS PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-219 000293 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 8): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 141. 82 . 105 . 227 . 134 . 104 . 97 . 14 5 . 103 . 116 . 114 . 148. 69 . 95. 79. 84 . 81. 190. 117 . 115. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 0.4 MG/KG PFOS 418-018:PAGE B-220 000294 PROTOCOL 418-018; ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER; T-6295.25) TABLE 31 (PAGE 9) ; MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 9 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 56 . 103 . 112 . 91. 95 . NOT PREGNANT 78 . 85 . NOT PREGNANT 97 . NOT PREGNANT 98 . 85 . 141. 127 . 126 . 88 . 87 . 115 . 45 . ALL WEIGHTS WERE RECORDED IN GRAMS (G),. DAYS = DAYS OF LACTATION 0.8 MG/KG PFOS 418-018:PAGE B-22 000295 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 10): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT tf DOSAGE GROUP 10 1 MG/KG PFOS 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 108 . 126 . 147 . NOT PREGNANT 77 . 108 . 217. 93 . 92 . 80 . 60 . 83 . 84 . 78 . 109. 87. 91. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 87. 107 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-222 000296 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 11) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 11 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 103 . NOT PREGNANT 98 . 85. 86 . 74 . 69 . 128 . 84 . 135 . 85 . NOT PREGNANT 72 . 131. 102 . 65 . 108 . 87 . 96. 101. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 1.2 MG/KG PFOS 418-018:PAGE B-22 000297 OJ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABI.E 31 (PAGE 12) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT tf DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11041 11042 11043 11048 11049 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 NOT PREGNANT 75 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS NOT PREGNANT SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 75 . 112 . 97. 85 . 45 . 91. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 100 . 46 . 68 . 43. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 87. 105 . NOT PREGNANT 57. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 13): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAGE GROUP 13 2 MG/KG PFOS 11055 11057 11059 11061 11064 11065 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 95. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 55 . 94 . 95 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 34 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 39. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS NOT PREGNANT SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-225 000299 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 1): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 V EH IC LE CONTROL RAT # DAYS IN CO H AB ITATION M ATING STATUS M ATING DATE 10901 2 Mc 10902 10903 10904 2 Mc 3 Mc 1 Mc 10905 10906 1 Mc 3Mc 10907 4 M c 10908 10909 1 Mc 3Mc 10910 3Mc 10 9 11 3Mc 10912 i MC 10913 13 Mc 10914 4Mc 115 0 1 2Mc 11502 1 Mc 11503 4 Mc 11504 5 Mc 11505 5 Mc 11506 2 Mc 115 0 7 1 Mc 11508 3Mc 11509 1 Mc 115 10 115 11 115 12 115 13 1 Mc 1 Mc 1 Mc 1 Mc 115 14 1 Mc M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P P P P P P P P P P P NP P P P P p P p P P P P p P P p P 418-018`.PAGE B-226 000300 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 2): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 2 MEVALONIC ACID CONTROL RAT # DAYS IN COHABITATION MATING STATUS MATING DATE 10915 iM C 10916 2M C 10917 3M c 10918 4 M c 10919 4 M c 10920 2M c 10921 3M c 10922 6M c 10923 4M c 10924 4M c 10925 2M c 10926 1M c 10927 12 M c 10928 2M c 11515 2M c 11516 3M c 11517 3M c 11518 1M c 11519 iM c 11520 3M c 11521 2M c 11522 3M c 11523 1M c 11524 2M c 11525 1M c 11526 4M c 11527 1M c 11528 1M c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P P P P P p P P p P p p P p P P P p P P P NP P p P p P P Toeioo 418-018:PAGE B-227 PROTOCOL 4IB -018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 3): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID RAT ft DAYS IN COHABITATION MATING STATUS MATING DATE 10929 10930 10931 10932 10933 10934 10935 10936 10937 10938 10939 10940 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 1M 3M 3M 1M 2M 3M iM FOUND DEAD ON DAY 2 OF STUDY 3M 3M 3M 2M 3M 3M 4M 1M 1M 1M 1M 4M iM iM 1M 2M 1M 1M 1M 4M 2M c C C c c C c C C c c c C c c C C C c C c c C C C c c c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P p P P P P P P P NP P P p P p NP p P P p P P p P P p P p 418-018:PAGE B-228 000302 PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 4): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 4 2 MG/KG PFOS; + MEVALONIC ACID RAT it DAYS IN COHABITATION MATING STATUS MATING DATE 10943 1M c 10944 4MC 10945 3MC 10946 3M c 10947 2M c 10948 5M c 10949 3Mc 10950 3M C 10951 3Mc 10952 12 M c 10953 3M c 10954 1M c 10955 3M c 10956 4M c 11544 1M c 11545 1M c 11546 1M c 11547 3M c 11548 4M c 11549 2M c 11550 4 M c 11551 1M c 11552 1M c 11553 2 M c 11554 2Mc 11555 1M c 11556 2 M c 11557 4M c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS p p P p P P p p P P P P P p P P p P NP P P P P P P P P P 418-018:PAGE B-229 ( 000 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 5) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 5 CHOLESTEROL CONTROL RAT # DAYS IN COHABITATION MATING STATUS MATING DATE 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 3M 2M 3M 2M 3M 3M 3M 3M 1M 4M 14 M 4M 4M 3M 2M 4M 3M 3M MORIBUND SACRIFICED ON DAY 16 OF STUDY 2M 2M 1M 4M 2M 3M 2M 4M 4M C c C C C C c c c c c c c c c c c c C C c c c c c c c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P P P p P p p P P p P p P P P P NP P P P P NP p P P P p 418-018:PAGE B-230 000304 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 6) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL RAT It D A Y S IN COHABITATION MATING STATUS MATING DATE 10971 10972 10973 10974 10975 10976 10977 10978 10979 10980 10981 10982 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 3M 3M 1M 1M 1M 3M 3M 3M 3M 4M 1M iM 4M 4M 1M iM 2M 2M 1M 1M 2M 4M 1M iM 1M 1M 2M iM C C c C c c C C c C c C c c C C c c c C C c C c c C c c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P P P P P P P P P P P P P P P P P P P P P P P P P P P P 418-018:PAGE B-23 000305 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 7) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL RAT it DAYS IN COHABITATION MATING STATUS MATING DATE 10985 3M C 10986 3M c 10987 3M c 10988 3M C 10989 1M c 10990 4M C 10991 3M c 10992 4M c 10993 2M c 10994 3M C 10995 2 M c 10996 3M C 10997 2M c 10998 2Mc 11586 1M c 11587 1M C 11588 5M c 11589 2M C 11590 2 M C 11591 1M c 11592 13 M c 11593 1M C 11594 1M c 11595 iM c 11596 1M c 11597 1M c 11598 3M c 11599 4M c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P p P P p P p p P P p P P NP P P P P P P P P P P P P P P a te o o o 418-018:PAGE B-232 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 8) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 8 0.4 MG/KG PFOS RAT tt DAYS IN COHABITATION MATING STATUS MATING DATE 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 3M 3M 2M 4M 3M 3M 3M 4M 3M 4M 3M 2M iM 2M 2M 4M iM 3M 3M 1M C C c c C c c c c c c c c c c c c c c c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS p P P p P P p P P p P P p P p P p P p P 418-018:PAGE B-23 0003? UJ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 9): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 9 0.8 MG/KG PFOS RAT # 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 DAYS IN COHABITATION 3 4 2 3 4 14 4 1 3 5 1 1 1 2 2 4 3 1 4 i MATING STATUS M M M M M DID NOT MATE M M M M M M M M M M M M M M MATING DATE c C C c c - c c c c c c c c c c c c c c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P P P P P P P NP P NP p P P p P P P p P 418-018:PAGE B-234 0003M PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 10) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 10 RAT # 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 DAYS IN COHABITATION 14 3 1 6 3 i 5 4 i 3 2 i 1 4 2 i 2 3 i 2 M = MATED C = CONFIRMED P = PREGNANT 1 MG/KG PFOS MATING STATUS M M M M M M M M M M M M M M M M M M M M NP = NOT PREGNANT MATING DATE C c c c c c c c c c c c c c c c c c c c PREGNANCY STATUS P P P NP P P P P p p P p p P p P P P P P eeeooo 418-018:PAGE B-235 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 11): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 11 1.2 MG/KG PFOS RAT # 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 DAYS IN COHABITATION 4 14 4 2 2 1 2 3 1 3 2 2 1 2 1 1 1 1 2 1 MATING STATUS M DID NOT MATE M M M M M M M M M M M M M M M M M M MATING DATE c - C c C c C c c c c c C C c c c c c c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS P - P P P p P p p P P NP P P P P p P p P TCO 0O 418-018:PAGE B-236 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 12) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 12 1.6 MG/KG PFOS RAT # DAYS IN COHABITATION MATING STATUS MATING DATE 11039 11040 11041 11042 11043 11044 11045 11046 11047 11048 11049 11050 11051 11052 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 14 1 4 4 2 1 1 3 3 4 5 1 3 1 3 3 2 1 2 1 1 1 4 i 13 4 3 1 DID NOT MATE M M M M M M M M M M M M M M M M M M M M M M M M M M M - c c c c c c c c c c c c c c c c c c c c c c c c c c c M = MATED C = CONFIRMED P = PREGNANT NP = NOT PREGNANT PREGNANCY STATUS - P P P NP p p p P p P P P P P p P p P P p P P P p P NP P 418-018:PAGE B-237 000311 PROTOCOL 4X8-018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 32 (PAGE 13): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 13 RAT # DAYS IN COHABITATION 11053 11054 11055 11056 11057 11058 11059 11060 11061 11062 11063 11064 11065 11066 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 M = MATED 4 4 3 4 i 3 4 1 3 1 2 12 1 2 1 3 2 2 3 2 3 2 1 1 2 1 3 3 c = CONFIRMED P = PREGNANT 2 MG/KG PFOS MATING STATUS M M M M M M M M M M M M M M M M M M M M M M M M M M M M NP = NOT PREGNANT MATING DATE C C c c c c c c c c c c c c c c c c c c c c c c c c c c PREGNANCY STATUS P P p P P p P P P P P p p P P P P P P p P P P P P P NP P 418-018:PAGE B-238 000312 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 33 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA RAT # RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP 1 VEHICLE CONTROL 10901 10902 10903 10904 10905 10907 10908 10910 10912 6 7 13 0 0 0 1 0 1 0 0 0 7 7 14 8 7 15 5 7 12 0 0 0 0 1 1 0 0 0 5 8 13 6 8 14 10 4 14 0 0 0 1 0 1 0 0 0 11 4 15 12 4 16 9 5 14 0 0 0 0 1 1 0 0 0 9 6 15 9 7 16 5 8 13 0 0 0 1 0 1 0 0 0 6 8 14 8 a 16 9 7 16 0 0 0 0 1 1 0 0 0 9 8 17 9 10 19 8 6 14 0 0 0 1 0 1 0 0 0 9 6 15 9 6 15 7 5 12 0 0 0 2 2 4 0 0 0 9 7 16 10 8 18 NOT PREGNANT DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10915 10916 10917 10918 10919 10920 10925 10928 9 5 14 0 0 0 i 1 2 0 0 0 10 6 16 12 7 19 7 9 16 0 0 0 0 1 1 0 0 0 7 10 17 7 11 18 7 3 10 0 0 040 40 0 0 11 3 14 ii 5 16 5 9 14 0 0 0 0 0 0 0 0 0 5 9 14 8 ii 19 6 9 15 0 0 0 0 0 0 0 0 0 6 9 15 ii 14 25 6 3 9 0 0 0 5 1 6 0 0 0 11 4 15 14 9 23 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 8 9 17 7 8 15 0 0 0 1 0 1 0 0 0 8 8 16 8 8 16 DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10930 10931 10932 10933 10934 10935 10937 10940 8 5 13 0 0 0 0 1 1 0 1 1 8 7 15 11 8 19 8 5 13 000 10 100 0 9 5 14 9 5 14 7 8 15 0 0 0 0 1 1 0 0 0 7 9 16 11 11 22 7 9 16 0 0 0 2 0 2 0 0 0 9 9 18 9 9 18 5 4 9 0 0 0 5 3 8 0 0 0 10 7 17 13 10 23 6 6 12 0 0 0 0 0 0 0 0 0 6 6 12 6 6 12 7 4 ii 0 0 0 2 0 2 0 0 0 9 4 13 9 4 13 7 12 19 00 000 00 0 0 7 12 19 7 12 19 PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-018:PAGE B-239 000313 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 33 (PAGE 2): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA RAT (t RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10945 10946 10947 10949 10950 10951 10953 10954 10 6 16 0 0 0 0 0 0 0 0 0 10 6 16 11 6 17 11 4 15 0 0 0 1 0 1 0 0 0 12 4 16 12 6 18 10 5 15 0 0 0 0 0 0 0 0 0 10 5 15 11 5 16 7 1 8 0 0 0 2 7 9 0 0 0 9 8 17 11 10 21 9 5 14 0 0 0 i 1 2 0 0 0 10 6 16 10 7 17 5 9 14 0 0 0 3 1 4 0 0 0 8 10 18 10 13 23 9 5 14 0 0 0 1 0 i Q 0 0 10 5 15 10 5 15 7 6 13 0 0 0 1 1 2 1 0 1 9 7 16 9 10 19 DOSAGE GROUP 5 CHOLESTEROL CONTROL 10958 10959 10960 10961 10962 10963 10964 10965 7 7 14 0 0 0 0 0 0 0 0 0 7 7 14 8 7 15 9 9 18 0 0 0 0 1 1 0 0 0 9 10 19 10 10 20 7 7 14 0 0 0 0 1 I 0 0 0 7 8 15 11 8 19 ii 5 16 0 0 0 0 0 0 0 0 0 11 5 16 11 5 16 5 8 13 0 0 0 1 0 I 0 0 0 6 8 14 8 12 20 11 7 18 0 0 0 0 0 0 0 0 0 11 7 18 11 8 19 7 10 17 0 0 0 1 0 1 0 0 0 8 10 18 10 10 20 9 3 12 0 00 10 10 0 0 10 3 13 10 3 13 DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10972 10974 10975 10976 10978 10980 10981 10982 7 10 17 0 0 0 1 0 1 0 0 0 8 10 18 11 12 23 7 11 18 0 0 0 0 0 0 0 0 0 7 ii 18 7 ii 18 5 1 6 0 0 0 0 0 0 0 0 0 5 1 6 6 6 12 6 5 11 0 0 0 2 1 3 0 0 0 8 6 14 8 8 16 10 7 17 0 0 0 0 0 0 0 0 0 10 7 *7 11 8 19 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 11 11 22 6 5 11 0 0 0 0 0 0 0 0 0 6 5 11 7 7 14 7 9 16 0 0 0 1 0 1 0 0 0 8 9 17 8 9 17 PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-018:PAGE B-240 000314 STCOOO PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 33 (PAGE 3): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA RAT # RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10985 10987 10988 10989 10990 10991 10992 10993 9 5 14 0 0 0 0 0 0 0 0 0 9 5 14 14 6 20 3 8 11 0 0 0 1 3 4 0 0 0 4 11 15 5 il 16 10 4 14 0 0 0 1 1 2 0 0 0 il 5 16 il 5 16 i2300000000012 3134 6 7 13 0 0 0 1 0 1 0 0 0 7 7 14 14 12 26 5 9 14 0 0 0 1 1 2 0 0 0 6 10 16 8 il 19 6 3 9 0 0 0 2 i 3 0 0 0 8 4 12 12 13 25 5 9 14 0 0 0 2 0 2 0 0 0 7 9 16 8 11 19 DOSAGE GROUP 12 1.6 MG/KG PFOS 11040 11044 11045 11046 11047 11050 11051 11052 3 5 8 0 0 0 0 1 1 0 0 0 3 6 9 7 6 13 9 5 14 0 0 0 0 0 0 0 0 0 9 5 14 9 10 19 5 9 14 0 0 0 1 0 1 0 0 0 6 9 15 6 10 16 6 8 14 0 0 0 2 1 3 0 0 0 8 9 17 8 9 17 0 10 10 0 0 0 0 1 1 0 0 0 0 11 11 0 11 il 6 7 13 0 0 0 0 1 1 0 0 0 6 8 14 7 8 15 9 4 13 0 1 1 0 0 0 0 0 0 9 5 14 9 5 14 6 11 17 0 0 0 0 0 0 0 1 1 6 12 18 6 12 18 DOSAGE GROUP 13 2 MG/KG PFOS 11053 11054 11056 11058 11060 11062 11063 11066 4 il 15 0 0 0 0 0 0 0 0 0 4 il 15 5 14 19 9 7 16 0 0 0 0 0 0 0 0 0 9 7 16 13 17 30 7 5 12 0 0 0 0 1 1 0 0 0 7 6 13 8 9 17 8 10 18 0 0 0 1 0 1 0 0 0 9 10 19 9 13 22 6 8 14 0 0 0 0 0 0 0 0 0 6 a 1.4 6 8 14 10 5 15 0 0 0 0 0 0 0 0 0 10 5 15 10 7 17 7 7 14 0 0 0 0 0 0 0 0 0 7 7 14 9 7 16 6 9 15 0 0 0 0 1 1 0 0 0 6 10 16 6 10 16 PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-018:PAGE B-2 4^ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-S295.25) TABLE 34 (PAGE 1) : LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS NUMBER OF LIVE FETUSES POOLED FETAL BODY WEIGHT (G) RAT ( TOTAL TOTAL DOSAGE GROUP i 10901 10902 10903 10904 10905 10907 10908 10910 10912 13 12 14 14 13 16 14 12 NOT PREGNANT 67.36 62.03 74.73 71.12 68.98 77.14 72.29 68.52 DOSAGE GROUP 2 10915 10916 10917 10918 10919 10920 10925 10928 14 16 10 14 15 9 16 15 76.48 89.44 53 .12 84.41 68.96 45.39 77.64 71.56 DOSAGE GROUP 3 10930 10931 10932 10933 10934 10935 10937 10940 13 13 15 16 9 12 11 19 62.00 65.26 74.42 82.94 40.86 64.44 55.49 86.00 --- CONCEPTUSES -----DEAD OR RESORBED N N% VEHICLE CONTROL 14 1 7.1 13 1 7.7 15 1 6.7 15 1 6.7 14 i 7.1 17 1 5.9 15 1 6.7 16 4 25.0 MEVALONIC ACID CONTROL 16 2 12.5 17 1 5.9 14 4 28.6 14 0 0.0 15 0 0.0 15 6 40.0 16 0 0.0 16 1 6.2 1.6 MG/KG PFOS + MEVALONIC ACID 15 2 13.3 14 1 7.1 16 i 6.2 18 2 11.1 17 8 47.0 12 0 0.0 13 2 15.4 19 0 0.0 sveoeo 418-018:PAGE B-242 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 34 (PAGE 2): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS NUMBER OF LIVE FETUSES RAT ft TOTAL DOSAGE GROUP 4 10945 10946 10947 10949 10950 10951 10953 10954 16 15 15 a 14 14 14 13 DOSAGE GROUP 5 10958 10959 10960 10961 10962 10963 10964 10965 14 18 14 16 13 18 17 12 DOSAGE GROUP 6 10972 10974 10975 10976 10978 10980 10981 10982 17 18 6 11 17 16 11 16 POOLED FETAL BODY WEIGHT (G) TOTAL 72.48 82.52 76.37 29.06 74.80 73.68 75.62 69.92 74.35 87.28 66.68 82.33 73.21 94.52 93.22 63.36 89.27 89.02 23.23 57.91 80.99 74.99 60.24 89.48 -- CONCEPTUSES DEAD OR :RESORBED N N% 2 MG/KG PFOS + MEVALONIC ACID 16 0 0.0 16 1 6.2 15 0 0.0 17 9 52.9 16 2 12.5 18 4 22.2 15 i 6.7 16 3 18.8 CHOLESTEROL CONTROL 14 0 0.0 19 1 5.3 15 1 6.7 16 0 0.0 14 1 7.1 18 0 0.0 18 1 5.6 13 1 7.7 1.6 MG/KG PFOS + CHOLESTEROL 18 1 5.6 18 0 0.0 6 0 0.0 14 3 21.4 17 0 0.0 16 0 0.0 11 0 0.0 17 1 5.9 TEOOtt 418-018 :PAGE B-243 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 34 (PAGE 3): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS NUMBER OF LIVE FETUSES RAT # TOTAL DOSAGE GROUP 7 10985 10987 10988 10989 10990 10991 10992 10993 14 11 14 3 13 14 9 14 DOSAGE GROUP 12 11040 11044 11045 11046 11047 11050 11051 11052 8 14 14 14 10 13 13 17 DOSAGE GROUP 13 11053 11054 11056 11058 11060 11062 11063 11066 15 16 12 18 14 15 14 15 POOLED FETAL BODY WEIGHT (G) TOTAL 70.04 53,47 67.85 17.94 68.09 64.65 41.10 66.19 40.71 69.56 76.36 72.10 47.25 71.08 61.99 88.06 78.30 71.53 55.25 82.23 76.60 75.77 74.99 83.80 -- CONCEPTUSES DEAD OR RESORBED N N% 2 MG/KG PFOS + CHOLESTEROL 14 0 0.0 15 4 26.7 16 2 12.5 3 0 0.0 14 i 7.1 16 2 12.5 12 3 25.0 16 2 12.5 1.6 MG/KG PFOS 9 i 11.1 14 0 0.0 15 i 6.7 17 3 17.6 ii 1 9.1 14 1 7.1 14 1 7.1 18 1 5.6 2 MG/KG PFOS 15 0 0.0 16 0 0.0 13 1 7.7 19 1 5.3 14 0 0.0 15 0 0.0 14 0 0.0 16 1 6.2 sisoeo 418-018:PAGE B-244 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 35 (PAGE 1): FETAL VITAL STATUS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP 1 VEHICLE CONTROL RAT ft CLs 10901 8/ 7 A A A E A A A / A A A A A A A 10902 6/ 8 A A A A A / A A A A E A A A 10903 12/ 4 A A A A A A A A A A E / A A A A 10904 9/ 7 A A A A A A A A A/ A A A A A E 10905 8/ 8 E A A A A A / A A A A A A A A 10907 9/10 A A A A A A A A A / E A A A A A A A 10908 9/ 6 E A A A A A A A A / A A A A A A 10910 10/ 8 A E A A A A A E A / A E A E A A A 10912 NOT PREGNANT DOSAGE: GROUP 2 MEVALONIC ACID CONTROL RAT ff CLs 10915 12/ 7 E A A A A A A A A A / A A A A A E 10916 7/11 A A A A A A A / A A A E A A A A A A 10917 11/ 5 E A E A E A A A A E A / A A A 10918 8/11 A A A A A / A A A A A A A A A 10919 11/14 A A A A A A / A A A A A A A A A 10920 14/ 9 A E A A E E A E A E A/ A E A A 10925 8/ 9 A A A A A A A A / A A A A A A A A 10928 8/ 8 A A E A A A A A / A A A A A A A A DOSAGE: GROUP 3 1. 6 MG/KG PFOS + MEVALONIC ACID RAT # CLs 10930 11/ 8 10931 9/ 5 10932 11/11 10933 9/ 9 10934 13/10 10935 6/ 6 10937 9/ 4 10940 7/12 A A A A E A A A A = ALIVE D = DEAD A A A A A A A/ L E A A A A A A A E A A A A A/ A A A A A A A A A A A/ E A A A A A A A A A A E A A E A A/ A A A A A A A A A E E E A E A A A A/ E E A E A A A A A A A A/ A A A A A A A A A A E A E A/ A A A A A A A A A A/ A A A A A A A A A A A A E = EARLY RESORPTION L = LATE RESORPTION /" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY 418-018:PAGE B-245 000319 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-295.25) TABLE 35 (PAGE 2): FETAL VITAL STATUS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS FETUS # 1 2 3 4 5 6 7 8 9 10 il 12 13 14 15 16 17 18 DOSAGE: GROUP 4 2 MG/KG :PFOS; + MEVALONIC ACID RAT # CLs 10945 11/ 6 A A A A A A A A A A / A A A A A A 10946 12/ 6 A A E A A A A A A A A A / A A A A 10947 11/ 5 A A A A A A A A A A / A A A A A 10949 11/10 E A A A A A A A / E E E E E E A E 10950 10/ 7 A A A A E A A A A A/ A A A A A E 10951 10/13 A E A E A A A E / A A A A E A A A A A 10953 10/ 5 E A A A A A A A A A / A A A A A 10954 9/10 L A A E A A A A A / A E A A A A A DOSAGE GROUP 5 CHOLESTEROL CONTROL RAT # CLs 10958 8/ 7 A A A A A A A / A A A A A A A 10959 10/10 A A A A A A A A A / E A A A A A A A A A 10960 11/ 8 A A A A A A A / A A A A A A A E 10961 11/ 5 A A A A A A A A A A A / A A A A A 10962 8/12 A E A A A A / A A A A A A A A 10963 11/ 8 A A A A A A A A A A A / A A A A A A A 10964 10/10 A A A E A A A A / A A A A A A A A A A 10965 10/ 3 A A A E A A A A A A / A A A DOSAGE GROUP 6 1 .6 MG/KG PFOS + CHOLESTEROL RAT # CLs 10972 11/12 10974 7/11 10975 6/ 6 10976 8/ 8 10978 11/ 8 10980 11/11 10981 7/ 7 10982 8/ 9 A A A A A A A A A = ALIVE D = DEAD E A A A A A A/ A A A A A A A A A A A A A A A A/ A A A A A A A A A A A A A A A/ A A A E E A A A/ A E A A A A A A A A A A A A A/ A A A A A A A A A A A A A A/ A A A A A A A A A A A A A/ A A A A A A A A E A A A/ A A A A A A A A A E =: EARLY RESORPTION L = LATE RESORPTION 7 " DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY 02T000 418-018'.PAGE B-246 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 35 (PAGE 3): FETAL VITAL STATUS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL RAT # CLs 10985 14/ 6 A A A A A A A A A / A A A A A 10987 5/11 A A E A / A A A A A A A A E E E 10988 11/ 5 A A A A E A A A A A A / A A A A E 10989 1/ 3 A/ A A 10990 14/12 A A A E A A A / A A A A A A A 10991 8/11 A A A E A A / A A A A A A A A A E 10992 12/13 A A A A E A A E / E A A A 10993 8/11 E A A E A A A / A A A A A A A A A DOSAGE GROUP 12 1. 6 MG/KG PFOS RAT # CLs 11040 7/ 6 A A A/ E A A A A A 11044 9/10 A A A A A A A A A / A A A A A 11045 6/10 A A A A E A / A A A A A A A A A 11046 8/ 9 E A A A A A E A / A A A A A A A A E 11047 0/11 / A A A A A A A A A E A 11050 7/ 8 A A A A A A / A A A A A A A E 11051 9/ 5 A A A A A A A A A / D A A A A 11052 6/12 A A A A A A / A A A L A A A A A A A A DOSAGE; GROUP 13 2 MG/KG PFOS RAT ft CLs 11053 5/14 A A A A / A A A A A A A A A A A 11054 13/17 A A A A A A A A A / A A A A A A A 11056 8/ 9 A A A A A A A / A E A A A A 11058 9/13 E A A A A A A A A / A A A A A A A A A A 11060 6/ 8 A A A A A A / A A A A A A A A 11062 10/ 7 A A A A A A A A A A / A A A A A 11063 9/ 7 A A A A A A A / A A A A A A A 11066 6/10 A A A A A A / A A E A A A A A A A A = ALIVE: D = DEAD E = EARLY RESORPTION L = LATE RESORPTION " / " DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY 418-018:PAGE B-247 000321 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 1) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 VEHICLE CONTROL RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 2 34 MF MF MF 5 MF 10906 10909 10911 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 22 23 23 23 23 23 23 23 23 22 23 23 23 23 23 23 23 23 23 16 14 17 17 10 11 15 15 15 17 15 10 11 8 15 14 14(1) 11 15 0 1 0 0 0 1 1 0 0 0 0 0 2 0 0 0 0 0 0 16 15 17 17 10 12 16 15 15 17 15 10 13 8 15 14 14 il 15 10 e 68 89 6 11 37 74 96 69 4 il 98 96 91 83 26 96 77 49 74 69 96 68 89 6 10 37 74 96 69 4 11 97 96 91 83 26 96 77 49 74 69 96 68 89 6 10 37 74 96 69 4 10 97 96 91 83 26 96 77 49 74 69 96 68 88 6 10 37 74 96 69 4 10 97 96 91 83 26 96 77 49 74 69 96 68 88 69 37 74 96 69 4 10 97 96 91 83 26 96 77 49 74 69 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 17 15 19 17 12 12 16 15 15 17 16 10 14 12 15 16 14 12 15 418-018:PAGE B-248 000322 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 2) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 2 MEVALONIC ACID CONTROL RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN i MF NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 10921 10922 10923 10924 10926 10927 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 22 14 0 23 13 0 23 14 0 23 13 0 MORIBUND SACRIFICED ON[ DAY 22 17 0 22 12 0 23 14 2 22 15 0 23 12 0 22 14 (1) 0 23 13 0 23 7 0 NOT PREGNANT 22 16 0 23 11 0 23 13 0 23 12 1 23 11 0 23 13 0 14 9 13 8 14 11 13 8 12 OF GESTATION; 17 6 12 7 16 6 15 6 12 9 14 6 13 6 74 16 8 11 5 13 9 13 8 11 7 13 7 5 5 3 5 DEATH il 5 8 9 3 7. 7 3 8 6 4 4 4 6 95 95 95 95 85 75 75 75 11 3 il 3 11 3 il 3 85 85 85 85 WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 6 11 6 11 6 il 6 11 75 75 75 75 6e 68 68 68 69 69 69 69 .9 3 93 93 93 67 67 67 67 67 67 67 67 43 33 33 33 88 56 94 84 74 76 88 56 94 84 74 76 88 56 94 84 74 76 88 56 94 84 74 76 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 16 16 14 14 18 12 18 16 15 14 15 13 17 11 15 13 12 14 418-018-.PAGE B-249 000323 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 3) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 M1 F NUMBER OF LIVE PUPS AT COMPLETION OF' DAY POSTPARTUM 2 34 MF MF MF 5 MF 10929 10936 10938 10939 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 22 13 0 FOUND DEAD ON DAY 2 OF STUDY 22 15 (2) 0 NOT PREGNANT 22 11(2) 0 23 13 0 22 16(1) 0 23 13 2 NOT PREGNANT 22 12 (1) 2 22 10(8) 0 22 15(1) 1 22 14 0 23 9 0 23 13 0 22 12(6) 0 22 9 0 22 14 1 22 12 0 22 15 0 22 11(1) 0 13 15 11 13 16 15 14 10 16 14 9 13 12 9 15 12 15 11 76 76 54 67 87 67 65 259 68 45 94 42 45 95 75 87 37 65 42 13 67 75 55 -- 153 -- 34 94 32 21 11 -- 34 37 65 42 12 67 65 55 --- 43 -- 34 94 32 -- 11 -- 12 37 55 42 -- 66 55 55 --- 43 -- 34 94 32 -- 11 -- 11 37 55 42 -- 66 55 55 --- 43 -- 34 94 32 -- 11 -- 11 37 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 13 15 14 15 17 15 14 10 17 14 11 13 14 9 15 13 16 16 Y3000 418-018:PAGE B-250 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR 1S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 4) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N 10943 10944 10948 10952 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 11556 11557 22 22 22 22 22 22 22 23 23 22 NOT PREGNANT 22 21 22 22 22 22 22 22 22 LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 13(1) 13 13(10) 14 14 (7) 14 14 12 (3) 14 (14) 15 (2) 0 0 0 0 1 1 0 0 0 0 13 13 13 14 15 15 14 12 14 15 14(5) 14 (2) 12 (9) 13 (7) 17 17(2) 14 16(16) 15(15) 0 0 0 0 0 0 0 0 0 14 14 12 13 17 17 14 16 15 1 MF 84 10 3 12 77 25 77 86 45 -58 63 48 21 42 6 11 5 10 12 2 --- NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 2 34 MF MF MF 5 MF 1. 12 -2 ------24 _- -1 -1 ------22 -_ -1 -------21 - _ 1 -33 131 --1 2--- --1 -21 ------ ---2------ - _ 2 - M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 15 14 17 15 15 15 14 13 15 15 14 16 13 14 17 17 17 16 17 418-018:PAGE B-25 000325 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE S) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 5 CHOLESTEROL CONTROL RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 2 34 MF MF MF 5 MF 10957 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 23 11 0 ii 29 22 15 0 15 78 23 15 0 15 87 22 15 0 15 7 22 16 1 17 88 23 17 0 17 89 23 14 0 14 10 4 23 18 0 18 7 11 NOT PREGNANT 23 16 0 16 12 4 MORIBUND SACRIFICED ON DAY 16 OF STUDY 23 16 0 16 97 23 15 0 15 5 10 23 14 0 14 59 NOT PREGNANT 23 10 1 ii 55 23 14 0 14 86 23 16 0 16 88 23 12 0 12 75 23 ii 0 11 38 29 78 87 B7 87 89 10 4 7 11 12 4 97 4 10 59 55 86 88 75 38 29 78 87 87 87 88 10 4 7 ii 12 4 a7 4 10 59 55 86 88 75 38 29 78 87 87 87 88 10 4 7 il 12 4 87 4 10 59 55 86 88 75 38 29 78 87 87 87 88 10 4 7 11 12 4 87 4 10 59 55 86 87 75 38 M = MALE F = FEMALE a. Value appeared incorrectly recorded and was excluded from group averages and statistical analyses. TOTAL IMPLANTATIONS N 12 15 16 15 16a 17 14 18 16 16 15 14 14 14 16 14 15 418-018:PAGE B-252 000321 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 6) ; NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N 10971 10973 10977 10979 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 22 22 22 22 22 22 22 22 22 22 23 22 22 21 23 22 22 22 22 22 LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 7 13 15 (3) 14 (2) 15 16 10 13 13 (5) 13 (1) 12 16 13(1) 17(1) 16(1) 13(1) 12 14 15(2) 12 (1) 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 8 13 15 14 15 16 10 13 13 13 12 16 13 17 16 13 12 14 15 13 1 N1 F 34 76 75 84 12 3 79 64 49 26 84 48 88 66 97 96 66 39 3 11 2 11 65 NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 23 54 41 84 12 3 67 12 31 26 84 4B 166 13 76 -28 3 10 2 11 32 -- -1 41 84 93 ---24 84 48 -66 -2 64 -27 3 10 2 11 11 _- -41 74 93 ---24 84 48 -66 -2 64 -24 3 10 1 11 11 -- -41 74 93 ---24 84 48 -66 -2 64 -24 3 10 1 11 -- M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 15 14 15 16 15 16 10 13 14 13 14 16 13 17 17 15 12 15 15 13 msooo 418-018:PAGE B-253 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 7) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF S MF 10986 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 22 22 22 22 23 NOT PREGNANT 23 22 22 22 22 22 23 22 22 22 22 23 22 23 15 0 15 0 15 0 13 0 16 0 13 13 13 13(10) 16(1) 17(1) 15 13 (1) 16 0 14 10(2) 15 17(12) 1 0 1 0 0 0 0 0 0 0 0 0 0 0 15 15 15 13 16 14 13 14 13 16 17 15 13 16 HD 14 10 15 17 87 78 78 58 12 4 76 76 76 310 5 88 11 4 48 97 -' 10 4 44 ii 4 41 74 i44 -- ii 4 14 1- -- 223 -- 53 25 -1 -- 133 i1 -- -- -- 43 -- ii 4 ---- 1- --- 23 15 -1 --- 32 --- __ -- 33 -- ii 4 ---- 1- --- 23 15 -1 --- 32 --- --- 33 -- ii 4 ------- 23 1S ---- 32 --- M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. ( 1 = NUMBER OF PUPS IN WHICH CANNIBALIZATION AND/OR AUTOLYSIS PRECLUDED THE DETERMINATION OF VIABILITY. TOTAL IMPLANTATIONS N 17 15 15 15 17 14 16 15 13 17 17 17 14 16 9 15 16 15 18 418-018:PAGE B-254 000328 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 8) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 8 0.4 MG/KG PFOS RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 23 22 23 22 23 22 23 23 23 23 23 22 22 22 22 23 23 22 23 22 14 14 15 14 14 (2) 16 13 17 17 13 14 16 10 15 13 16 9 14 18 19(1) i 0 0 0 2 0 1 1 0 1 1 0 0 1 0 0 1 0 0 0 15 14 15 14 16 16 14 18 17 14 15 16 10 16 13 16 10 14 18 19 10 4 4 10 10 5 77 75 6 10 58 5 12 6 11 3 10 77 6 10 64 69 85 79 45 95 8 10 6 12 10 4 4 10 10 5 77 75 6 10 58 5 12 6 11 3 10 77 6 10 64 68 85 79 45 95 8 10 6 12 10 4 4 10 10 5 77 75 69 58 5 12 6 11 3 10 77 6 10 64 68 85 79 45 95 8 10 6 12 10 4 4 10 10 5 77 75 69 58 5 12 6 11 3 10 77 6 10 64 57 85 79 45 95 8 10 6 12 10 4 4 10 10 5 77 75 69 58 5 12 6 11 3 10 77 6 10 64 57 85 79 45 95 8 10 6 12 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM, TOTAL IMPLANTATIONS N 17 14 15 16 18 19 15 18 17 15 16 16 13 17 15 16 13 16 18 19 418-018:PAGE B-255 000329 PROTOCOL 418-018 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 9) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 9 0.8 MG/KG PFOS RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN N NN 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF1 DAY POSTPARTUM 234 MF MF MF 5 MF 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 23 22 23 23 22 NOT PREGNANT 22 22 NOT PREGNANT 23 NOT PREGNANT 22 23 22 23 23 22 22 23 22 13 19 16 16 16(1) 16 il 10 15 13 14 17 13 15 17 il 13 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 13 19 16 16 16 16 12 10 15 13 14 17 13 15 17 12 13 94 9 10 88 97 87 6 10 65 64 96 3 10 86 11 6 94 87 10 7 56 76 94 9 10 88 86 86 59 65 64 96 3 10 86 11 6 94 87 96 56 76 94 97 88 86 86 58 65 64 96 3 10 86 10 6 94 87 96 56 76 94 97 88 86 86 47 65 64 96 3 10 86 10 6 94 87 96 55 76 94 97 88 86 86 46 65 64 96 3 10 86 10 6 94 87 96 55 76 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 14 20 16 16 16 18 13 12 15 13 15 18 13 15 17 13 13 418-018:PAGE B-256 000330 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEI, INVESTIGATION IN RATS (SPONSOR1S STUDY NUMBER: T-629S.25) TABLE 36 (PAGE 10) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 10 1 MG/KG PFOS RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 22 23 23 NOT PREGNANT 22 23 24 22 22 22 22 22 22 22 23 22 23 22 22 22 14 15 17 16 9 17 13 16 20(1) 15 15 14 15(1) 14 17 14 15(1) 17 12(1) 0 14 0 15 0 17 0 0 0 0 0 0 0 0 0 0 2 0 0 0 o 0 16 9 17 13 16 20 15 15 14 15 16 17 14 15 17 12 77 B7 5 12 6 10 72 5 12 76 10 6 9 10 78 5 10 95 95 3 11 6 11 4 10 10 4 6 11 65 77 87 5 12 67 62 5 12 76 10 6 89 77 5 10 94 85 3 11 6 11 4 10 51 6 10 65 77 87 5 12 67 62 5 12 76 10 6 89 77 5 10 94 85 3 ii 6 11 4 10 -- 6 10 65 77 87 5 12 67 62 5 12 66 10 6 89 77 5 10 94 85 3 11 6 11 4 10 -- 6 10 65 67 87 5 12 67 62 5 12 66 10 6 89 77 5 10 94 84 3 ii 6 10 4 10 -- 6 10 65 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 15 15 17 16 11 18 16 17 20 15 16 14 17 17 18 15 16 17 13 Tetovo 418-018:PAGE B-257 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR1S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 11) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 11 1.2 MG/KG PFOS RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 MF NUMBER OF LIVE COMPLETION OF` DAY 23 MF MF PUPS AT POSTPARTUM 4 MF 5 MF 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 22 NOT PREGNANT 23 22 23 23 22 23 22 22 22 NOT PREGNANT 23 22 22 22 22 22 23 22 16 (1) 13 13 14 13 11(1) 17 16(3) 19 7 9 16 13 14 16 15(1) 14 16 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 16 13 13 15 13 11 17 16 19 7 9 16 13 14 16 15 14 16 69 94 94 4 10 49 64 12 5 3 10 9 10 34 54 97 67 10 4 97 77 59 88 69 94 94 49 39 64 12 5 3 10 89 34 32 97 67 64 97 77 59 77 69 94 63 49 28 s4 12 5 3 10 66 34 32 97 67 54 97 77 49 77 69 84 52 48 27 44 10 5 3 10 56 34 32 97 67 53 97 77 49 77 69 84 52 48 i7 44 10 5 3 10 46 34 32 97 67 53 97 77 49 77 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 16 17 16 16 15 14 18 17 19 7 13 16 14 15 16 16 14 16 000 418-018:PAGE B-258 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR1S STUDY NUMBER: T-6295.25) TABLE 36 (PAGE 12) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 12 1.6 MG/KG PFOS RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF1 DAY POSTPARTUM 2 34 MF MF MF 5 MF 11039 11041 11042 11043 11048 11049 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 NOT PREGNANT 22 22 NOT PREGNANT 22 22 22 22 22 22 22 22 22 22 22 22 22 22 NOT PREGNANT 22 17 16(2) 15(1) 6(1) 16 15(2) 16 12 14 (1) 10 11 11 13 (2) 17(1) 13 11(3) 14 0 0 0 0 0 0 0 0 0 4 0 0 0 0 0 0 0 17 16 15 6 16 15 16 12 14 14 11 11 13 17 13 11 14 6 11 77 77 32 88 58 88 48 85 64 38 47 56 88 67 53 95 6 10 -- -- 2i 88 46 88 15 74 -- 37 -2 46 31 57 52 85 -7 -- -21 88 46 88 13 74 -- 37 -1 46 257 52 84 -5 -- -21 78 46 88 i2 74 -- 37 -i 46 257 42 84 -5 -- -21 78 46 88 -2 64 -- 37 -1 46 -- 57 42 84 M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 17 16 15 10 15 15 17 12 14 15 12 13 15 18 13 12 14 418-018:PAGE B-259 000333 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 36 (PAGE 13) : NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 13 2 MG/KG PFOS RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) N 11055 11057 11059 I1Q61 11064 11065 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 22 22 23 22 22 22 22 23 23 22 22 22 22 22 22 22 22 22 NOT PREGNANT 22 LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 14(1) 10(1) 15 16 10 14 14 14 12(2) 13 (8) 14(12) 6(6) 14 15(12) 13 16(2) 11(2) 16 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 14 10 15 16 10 14 14 14 12 13 14 6 14 15 13 16 11 16 14 (10) 1 15 1 MF 58 45 11 4 79 73 68 59 4 10 46 32 -2 -- 95 12 94 86 63 6 10 22 NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 22 2_ _ __ -- 11 2 22 1- -- 54 48 46 1- -- ii 2 -_- -- 44 48 46 -- -- 11 2 --- -- 44 47 46 -- -- 11 2 -- -- 44 47 46 -- -- -- -- -- -- 12 -- 21 ---- -1 ---- -1 ---- -1 -- -- -- -- -- 23 -- i1 -- 11 --- -- -- -- -- M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 15 ii 18 16 11 14 14 15 12 16 15 14 15 15 13 16 12 17 15 418-018 :PAGE B-260 000334 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 1) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 1 VEHICLE CONTROL RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10906 10909 10911 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 94 .8 87 ,1 108 .0 112 .3 61..4 73 .1 96 .6 91..4 97..7 94 .3 103 .1 73 .2 79 .7 53 .9 92 ,0 92 ,.0 77 ,.6 71,.0 92 .7 93 .,2 94 .9 I l l .2 107 ,.9 65 .4 77 .7 107..6 102 .1 106 .8 105 .2 113 .4 86 .6 86 .8 61 .8 97 .3 102 .2 87 .1 79 .9 106 .8 106 .5 103 ,9 117 .8 113 .,6 73 .,2 83 .4 121..3 113 .0 115 ,.0 119.,2 127 .8 91 .5 99,.5 71,.9 107 .1 104 .1 96 .4 89 .2 118 .9 121 .1 121. 6 123 .1 123 .5 80. 5 93 .3 133.,5 122 .4 135 .,6 135 .4 148 .0 103 ,0 112 ,4 84 .5 119 .7 114 ,.0 108 .5 99 .0 .135 ,5 136..0 132 ,.5 131,.7 124 .0 92 .7 106 .0 148 ,.9 136 .5 155 .9 151..8 166 .6 116 .2 127 .1 96 .7 134 .0 127 .2 117 .5 I l l .1 146 .5 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-2 000335 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 2) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 2 MEVALONIC ACID CONTROL RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10921 10922 10923 10924 10926 10927 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 84.2 92.0 80.3 88.9 89.9 96.5 87.0 91.0 MORIBUND SACRIFICED ON DAY 95.7 103.3 74.0 82.2 93.0 102.8 90.3 96.7 79.0 90.1 73.8 80.5 90.1 93.2 47.0 54 .4 NOT PREGNANT 94.4 103.5 79.2 88.2 85.8 91.9 80.7 87.3 75.9 84.3 93.6 104.9 99.6 92.6 107.3 95.4 12 OF GESTATION; 116.1 93 .2 116.6 108.7 97.7 89.5 101.5 53.6 117.1 97.8 103.0 94.6 97.5 112.2 DEATH WAS 115.9 107.3 119.5 103.1 ATTRIBUTED TO AN 131.5 107.1 130.0 121.8 106.7 101.4 115.1 63.0 131.5 110.6 112 .3 107.0 109.8 123 .9 122.0 119.9 132.7 108.0 INTUBATION ACCIDENT 144.9 120.6 146.7 134.7 117.3 110.8 129.9 71.8 145.3 119.7 126.4 123 .6 124 .1 132.5 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-262 000336 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 3) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10929 10936 10938 10939 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 72.1 62.6 FOUND DEAD ON DAY 2 OF STUDY 68.0 34 .5 NOT PREGNANT 46.7 20.5 83.7 84.6 74.0 59.9 79.3 63.9 NOT PREGNANT 60.8 NO SURVIVING PUPS 11.0 5.4 71.8 40.1 72.4 NO SURVIVING PUPS 61.1 49.2 84.5 86.2 32.1 28.1 49.4 14.8 70.7 9.9 62.9 NO SURVIVING PUPS 77.2 31.2 57.5 63.7 65.0 38.5 14.2 89.0 62.0 70.0 67.8 75.5 44.8 47.9 NO SURVIVING 95.1 64.5 77.9 PUPS 104.3 71.6 85.4 NO SURVIVING PUPS 37.1 42.4 55.5 93.4 31.2 NO SURVIVING 9.6 PUPS 63 .0 135.8 35.2 10.5 12.0 74.3 7.8 82.1 47.8 71.5 109.0 40.5 11.2 7.1 93 .2 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-263 000337 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 7 (PAGE 4) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10943 10944 10948 10952 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 11556 11557 64.3 67.4 14.2 68.4 36.8 69.9 77.6 46.4 NO SURVIVING PUPS 68.1 NOT PREGNANT 49.0 63.9 16.1 33.3 91.0 69.1 77.6 NO SURVIVING PUPS NO SURVIVING PUPS 20.2 13.7 8.3 NO SURVIVING PUPS NO SURVIVING PUPS NO SURVIVING PUPS NO SURVIVING PUPS NO SURVIVING PUPS 31.1 NO SURVIVING PUPS 29.1 5.2 23.1 NO SURVIVING PUPS a 9.6 NO SURVIVING PUPS 4.6 4.0 21.3 4.6 NO SURVIVING PUPS 20.7 NO SURVIVING PUPS NO SURVIVING PUPS 3.9 NO SURVIVING PUPS 15.8 NO SURVIVING PUPS 16.6 NO SURVIVING PUPS 4 .7 17.3 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS, a. Value was not recorded. 418-018:PAGE B-264 000338 PROTOCOL 418-018: OWE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 5) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 5 CHOLESTEROL CONTROL RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10957 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 71.9 75.5 82.9 97.0 106.9 127.2 100.0 105.8 115.8 83.5 88.5 93.7 90.2 88.6 95.1 97.8 105.3 106.4 90.6 101.6 111 .9 110.9 120.6 135.8 NOT PREGNANT 95.1 105.0 123.6 MORIBUND SACRIFICED ON DAY 16 OF STUDY 106.4 117.5 122.9 89.2 96.5 105.0 97.2 107.6 119.0 NOT PREGNANT 64.9 73.4 83.5 85.1 91.0 100.0 106.6 153.0 134.6 79.6 87.8 92.7 79.9 87.0 98.3 97.9 143.1 136.7 109.7 110.2 121.1 124.6 152.3 140.6 140.4 113 .4 132.9 98.3 1 1 1 .9 147.7 104.3 113 .7 106.1 163.6 142.1 120.3 117.5 130.3 137.4 173.5 157.2 152.3 127.0 148.1 107.6 125.0 a 110.9 131.0 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. a. Value was not r e c orded. 418-018:PAGE B-265 060339 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 6) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10971 10973 10977 10979 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 35.2 69.1 59.9 60.7 82.9 77.1 55.5 70.5 40.1 72.7 72.6 89.7 69.1 79.9 89.1 61.4 64.9 76.7 69.0 58.6 22.3 44.4 26.8 64.6 75.9 55.5 14.6 19.8 39.9 79.3 74.6 5.3 72.8 17.7 76.1 NO SURVIVING PUPS 49.4 71.7 72.4 25.2 NO SURVIVING 4.2 29.7 71.8 68.6 NO SURVIVING NO SURVIVING NO SURVIVING 33 .1 91.8 81.6 NO SURVIVING 81.8 9.2 60.2 PUPS PUPS PUPS PUPS PUPS NO SURVIVING 36.4 77.0 78.0 36.5 105.1 94.0 92.4 10.1 65.6 PUPS 39.8 83.3 85.2 40.0 112.9 102.2 99.6 10.0 72.2 46.0 76.6 79.8 10.3 38.4 86.1 83 .2 9.5 44 .5 95.6 91.5 NO SURVIVING PUPS DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-266 000340 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 7) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10986 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 79.9 75.1 74.8 70.7 86.0 NOT PREGNANT 76.3 69.7 61.0 16.6 79.6 84.8 87.6 65.5 77.5 NO SURVIVING PUPS 76.3 47.0 83.8 23.6 52.1 4.4 40.4 NO SURVIVING 81.2 PUPS 25.4 4.3 NO SURVIVING PUPS 11.0 22.7 NO SURVIVING PUPS 41.9 35.0 4.8 4.5 36.8 9.2 NO SURVIVING PUPS NO SURVIVING PUPS NO SURVIVING PUPS 3B.6 85.9 NO SURVIVING PUPS NO SURVIVING PUPS 5.8 NO SURVIVING PUPS 27.4 31.9 4 .7 NO SURVIVING PUPS 33.8 NO SURVIVING PUPS 33.6 92.5 5.2 30.3 36.1 4 .5 35.1 34.1 94.6 NO SURVIVING PUPS 34.9 40.1 NO SURVIVING PUPS 35.7 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. twroeo 418-018:PAGE B-267 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 8) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 8 0.4 MG/KG PFOS RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 B8 .0 80. 9 59 .3 81. 6 77 ,4 86..1 84 .1 105 .6 103 .0 80 .7 92 .5 107 .6 57..7 85 ,i 71..9 92 .4 59..4 87 .7 107 .3 96 .6 93 .2 85 .,5 96 .5 87 .7 84 .0 82 .1 89 .2 110..8 109 .5 87 .5 93 .9 115 .8 61 .3 84 .4 76 .6 100 .7 66 .0 98 .3 113 .6 102 .4 100 .9 92 .8 96..7 101..i 91..8 77,.5 96 .3 119 .5 121 .0 97 .1 100 .7 131 .1 66 .0 85 .6 85 .9 109 .5 72 .1 109 .9 125 .8 109 .0 119 .5 106..8 112 .3 113 .1 107 .5 85 .0 109 .3 129 .2 135 .3 a 116 .1 147 .0 74 .1 82 .2 94 .6 110 .3 80 .7 122 .5 a 117 .5 129,.8 118 ..1 119 .,0 124 ,6 115 .,5 92 .,0 118 ,6 146 .4 144 .9 120 .6 132 .0 161 .4 81 .6 94 .2 101 .5 126 .5 90 .8 136 .2 147 .4 129 .7 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. a. Value was not recorded. ZfrCOOO 418-018-.PAGE B-268 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 9): PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 9 0.8 MG/KG PFOS RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 85.4 94.1 97.5 98.0 76.9 NOT PREGNANT 79.8 64.7 NOT PREGNANT 70.7 NOT PREGNANT 87.9 85.4 83.0 102.1 82.2 80.3 93.3 74.2 75.2 88 .0 95 .0 107 .1 90 .0 73 .,7 72 .4 68 .,5 78 ,3 94 .9 94 .0 96 .2 I l l ,5 93 .,9 84 .6 89 ,7 73 .5 80,,7 93 ,.8 84 ..7 Ill .3 99 ,5 78..6 69 .5 76 ,0 87 ,8 106,.2 102 ,.2 108 .3 119 .5 105 .4 94 .1 93 .7 81,.1 86 .6 101,,1 94 ,4 131,.0 115 .0 89 .3 64 .4 86 .2 100 .1 117 .2 111 .6 123 .0 132 .2 121 .2 103 .5 100 .8 86 .3 95 .0 101,.8 101,,6 139 ,.2 121..8 95 ,2 62 ,5 96 ,5 113 ,.4 125 ,.7 120 ,.9 135 ,.2 147 ,.9 133 .5 112 .3 109 .9 99 .6 98 .1 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-269 000343 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 10): PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 10 1 MG/KG PFOS RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 79.8 93.7 119.4 NOT PREGNANT 87.8 70.8 107.0 76.6 88.4 101.1 74.2 86.2 73.9 75.9 85.6 101.6 88.7 62.4 88.5 68.9 82.5 101.1 110.7 74.1 61.3 113.4 81.8 94.0 91.2 69.6 92.0 71.4 72.2 91.0 107.0 95.0 25.6 85.7 80.4 91.9 109.1 117.2 104.1 124 .4 131.2 81.9 68.8 124.9 91.9 99.1 96.1 73.9 103.0 77.7 73.9 100.2 113.0 104.9 NO SURVIVING 90.7 93.1 PUPS 92.7 80.5 137.9 90.1 109.3 108.4 80.0 114.2 87.7 78.5 1 1 1 .0 121.8 113.1 102.4 105.0 107.8 133.0 140.5 100.5 90.1 153.8 96.2 115.9 116.3 87.8 122.4 94.4 81.0 123.9 125.4 117.5 1 1 1 .0 116.3 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-270 000344 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 7 (PAGE 11) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 11 1..2 MG/KG PFOS RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 77. 7 NOT PREGNANT 79. 4 64. 0 89. 6 79 .8 50 .0 111. 6 74 .7 87 .5 40 .,2 NOT PREGNANT 59. 4 90. 7 74 ,9 80. 4 91 .9 79 .8 79 .,1 79 .,4 78 .9 84 .6 63 .0 86 .5 70. 8 49. 3 114 .2 73 .5 75 .3 48 .4 32. 4 97..2 83 .,7 52 ,.3 99..1 76 ,.6 84 .7 70 .2 84 .,3 93 .2 44 ,.9 83 .9 60 .,6 45 .1 119 ,i 80..8 55 ,1 56 .0 34 .1 111 .0 94 .6 45 .6 111 .5 81 .6 88 .4 78 .0 91..3 93 .7 43 .7 87 .7 55 .,2 39 .7 113 .2 88 .,i 50 .6 63 .6 37..7 119 .2 105 .9 43 .9 120 .8 87 .0 93 .0 90 .6 99 ,.8 100 .8 47 ,.3 90..1 49 ,0 40,.6 123 .7 97 .0 49..5 72 .6 41 .4 131 .i 116 .9 46 .8 133 .2 90 .5 105 .0 97 .6 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. o A IT 2 418-018-.PAGE B-2 -J PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 37 (PAGE 12) : PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 12 1..6 MG/KG PFOS RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 11039 11041 11042 11043 11048 11049 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 NOT PREGNANT 89.8 67.5 NOT PREGNANT 68.2 27.6 84.0 67.0 84 .3 68.2 66.5 52.9 77.5 57.5 55.1 82.6 70.4 43.9 NOT PREGNANT 75.6 77.9 33.2 NO SURVIVING PUPS NO SURVIVING 17.3 90.2 54.1 86.8 30.2 60.9 NO SURVIVING 67.0 8.6 51.6 20.3 66.8 40.1 PUPS PUPS 18.7 97.1 60.1 96.5 19.5 67.9 75.7 4.7 56.8 10.1 72.0 42.5 68.0 66.5 24.5 21.1 99.7 64.3 108.7 15.5 73.5 85.0 5.2 65.1 9.7 82.6 42.6 72.6 24.2 23.4 108.2 69.7 121.7 12.9 75.7 93.0 5.5 73.8 NO SURVIVING 90.1 48.7 PUPS 78.7 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS, 9 +1 $ O Q Q 418-018:PAGE B-272 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 37 (PAGE 13): PUP BODY WEIGHTS POOLED BY LITTER FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL D ATA - FI GENERATION LITTERS MATERNAL DOSAGE GROUP 13 2 MG/KG PFOS RAT/ LITTER NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 11055 11057 11059 11061 11064 11065 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 65.7 49.5 88.4 79.6 54.9 71.4 78.2 83.8 65.2 23.8 8.6 NO SURVIVING PUPS 75.3 14.4 71.2 72.5 49.1 82.1 NOT PREGNANT 19.4 19.6 NO SURVIVING PUPS 80.4 17.5 5.0 NO SURVIVING PUPS 46.6 72.9 70.2 4.4 NO SURVIVING PUPS 14.2 NO SURVIVING PUPS 14.9 NO SURVIVING PUPS NO SURVIVING PUPS 22.4 NO SURVIVING PUPS 9.6 86.0 NO SURVIVING PUPS NO SURVIVING PUPS 45.5 78.5 76.9 NO SURVIVING PUPS NO SURVIVING PUPS 5.1 9.5 NO SURVIVING PUPS 91.9 53.0 82.3 83.1 5.1 9.1 99.0 57.4 91.7 88.3 5.0 NO SURVIVING PUPS DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018 :PAGE B-273 *0 PROTOCOL 418-018; ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 1): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 1 2 3 4 5 6 7 8 9 10 li 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER # MATERNAL DOSAGE GROUP 1 VEHICLE CONTROL 10906 10909 10911 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA FA MA MA MA MA MA MA MA MA MA FA MA MA MA MA MA MA MA MA FA MA MA MA MA MA MA MA MA MA FA MA MA MA FA MA MA MA MA FA MA MA FA MA MA MA MA MA MA FA MA MA MA FA MA MA MA MA FA MA MA FA MA MA MA MS MA FA FA MA MA FA FA MA MA MA MA FA MA MA FA MA FA MA FA MA FA FA FA MA FA FA MA MA MA MA FA MA FA FA FA FA M A MM 2 FA FA FA FA FA FA FA FA FA FA MA FA FA FA FA FA MA FA MA FA MA FA FA FA MA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FS FA FA FA FA FA FS FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA 3 F A FM 5 ] FA FA FA FA FA FA FA FA FA F A FD 3 FA FA FA FA FS FA 3 FS FA FA FA FA F A FD i FA FA FA FA FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A - A L I V E , S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. frCQOO 418-018:PAGE B-274 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OP PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 2): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 12 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER # MATERNAL DOSAGE GROUP 2 MEVALONIC ACID CONTROL 10921 10922 10923 10924 10926 10927 11515 11516 11517 11518 11519 11520 11521 11522 11523 11524 11525 11526 11527 11528 MA MA MA MA MA MA MA MA MA FA FA FA FA FA M A M A M A M A M A M A M A MM 3 F A F A F A F A F A M A M A M A M A MA M A M A MA MA MA MA FA FA FA MA MA MA MA MA MA MA MA FA FA FA FA FA MORIBUND SACRIFICED ON DAY 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT MA M A MA MA MA M A FA FA FA FA FA FA FA FA FA FA FA MA MA MA MA MA M A MA FA FA FA FA FA M A M A M A M A M A MA M s FA FA FA FA FA FA FA FA FS MA M A M A MA M A M A FA FA FA FA FA FA FA FA FA MA M A MA MA MA MA MA MA MA FA FA FA M A M A M A M A M A M A F A F A F A F A F A F A F A FD 1 M A MA MA M A M A M A FA FA FA FA FA FA FA M A M A M A MM 3 F A F A F A NOT PREGNANT MA MA MA MA MA MA MA MA FA FA FA FA FA FA FA FA M A M A M A MA M A FA FA FA FA FA FA M A M A M A MA MA MA M A M A MA FA FA FA FA MA M A MA MA MA MA M A MA FA FA FA FA FS MA M A M A MA MA M A MA FA FA FA FA MA MA MA MA MA MA MA FA FA FA FA FA FA FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A - A L I V E , S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. &YEOOO 418-018:PAGE B-275 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 3): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 1 2 3 4 5 6 7 8 9 10 ii 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER ft MATERNAL DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 10936 10938 10939 10941 10942 11529 11530 11531 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 M A M A M A M A M A MD 4 MD 2 F A F A F A F A F A FD 2 FOUND DEAD ON DAY ;l OF STUDY M A M A M A M A MM 2 MM 2 MD 2 MD 1 F A F A FM 2 FM 2 FM 2 FM 2 FD 1 NOT PREGNANT MM 4 MM 2 MM 2 MM 2 MM 2 MD 1 MD 1 FM 4 FM 4 FM 3 FM 2 M A M A M A M A M A M A F A F A F A F A F A F A FM 4 M A M A M A M A M A MM 4 MM 3 MD 2 F A F A F A F A F A FD 2 FM 2 FD 1 M A M A M A M A M A MM 2 F A F A F A F A F A FD 2 FM 2 F S F S NOT PREGNANT MM 2 MM 2 MM 2 MM 2 MD 2 MD 2 M S M S FD 2 FD 2 FD 2 FD 2 FM 2 FD 1 MM 3 MM 2 MD 1 MD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 M A M A M A M A MM 3 M S F A F A F A FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 1 MD 2 MD 2 MD 2 MD 2 MD 2 MM 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 M A M A M A MM 2 F A F A F A F A FM 2 MA MA M A MA MA MA MA MA MA FA FA FA FA M A M A M A MD 2 MD 1 MD 1 MD 1 F A F A FD 1 FD 1 FD 1 MD 3 MD 3 MD 2 MD 2 FM 3 FD 2 FD 2 FD 2 FD 2 M A MD 2 MD 2 MD 2 MM 2 MM 2 MM 2 MM 2 MM 2 F A FD 2 FD 2 FM 2 FM 2 F S MD 2 MD 2 MD 2 MD 2 MD 2 MM 2 MM 2 FD 2 FD 2 FD 2 FD 2 FD 2 M A MM 3 MM 3 MD 2 MD 2 MD 2 MD 2 MD 2 F A FM 4 FD 3 FM 3 FD 2 FD 2 FD 2 M A M A M A F A F A F A F A F A F A F A FD I FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018.-PAGE B-276 000350 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 8 (PAGE 4) : PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER # MATERNAL DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 10944 10948 10952 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 11552 11553 11554 11555 11556 11557 MD 3 MD 2 MD 2 MD MD 2 MD 2 MD 2 MM MM 2 MD i MD 1 MD MD 2 MD 2 MD 2 MD MD 2 MD 2 MD 1 M MD 2 MD 2 MD 2 MD MD 2 MD 2 MD 2 MD MM 2 MM 2 MM 2 MM MD 1 MD i MD 1 MD MM 5 MM 5 MM 2 MM NOT PREGNANT MM 2 MM 2 MM 2 MM MD 3 MD 3 MM 3 MD MM 3 MM 2 MD 1 MD M A M A MD 3 MM MD 2 MD 2 MD 2 MD MD 2 MD 2 MD 2 MD MD 3 MD 3 MD 2 MD MD 1 MD 1 MD 1 MD MD 1 MD 1 MD 1 MD 2 MD 2 MM 1 MD 2 MD S FD 2 MD 2 MD 2 MD 1 MD 2 MM 2 MD 2 MD 1 MD 2 MD 2 MD 2 MM 2 MD 1 MD 1 MD 2 MD 2 MM 1 MD 2 MD 2 FD 2 MD 2 MD 1 FM 1 FD 2 MD 2 MD 1 FM 1 MD 1 MD 2 MD 2 FD 2 MD 1 MD 1 MD 2 MD 2 MD 2 FD 2 MM 2 MM 2 MM 1 FM 4 FM 3 FD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FM 2 MD 2 M S FM 2 MD 2 MD 2 FD 2 FM 2 FM 2 FD 1 FD 1 FD 1 FD 1 F A FM 4 FD 2 FM 2 FD 2 FD 4 FM 3 FM 3 FD i FM 2 FD 1 FD i MD i MD 1 FD 2 FD 2 FD 2 FD 3 FD 2 FD 2 FD 2 MD 2 MD 2 MD 1 MD 1 FD 1 FD 1 MD 1 FD 1 FD 2 FD 2 MM 1 FD 2 FD 2 FD 2 FD 2 FD 2 FD 1 FD 3 FM 2 FD 2 FD 1 FD 4 FM 2 FD 2 FD 2 MD i FD 1 FD 2 FD 2 FD 3 FM 2 FM 1 FD 1 FD 2 FD 2 FD 1 FD 1 FD 2 FD 2 FD 2 FD 2 FD 2 FD 1 FD 1 FD 1 FD 3 FD 2 FD 1 FD 1 FD 2 FD 2 FD 1 FD 1 FD 2 FD 1 FD 2 FD 2 FD 2 FD 2 FD 2 MD 2 MD 1 FD 1 FD 1 FD 1 FD 2 FD 5 FM i FD 2 FD 1 FD 2 FD 2 FD 1 1 FD 2 FD i FD 2 FD 1 1 FD 2 FD 2 FD 2 FD 1 FD 1 FD 1 3 1 2 FD 1 FD 2 FD 2 FD 1 FD 2 FM 1 FD 2 FD 1 2 FD 2 FD 2 FD i FD 1 FD 2 1 FD 2 FD 2 1 2 FD 1 1 2 FD 2 FD 2 1 FD 1 FD 1 2 1 2 FD 2 FD 1 FD 1 2 fd 1 fd 1 : : FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-277 000351 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE S) : PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP tt 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER # MATERNAL DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 M A M A FA FA FA FA FA FA FA FA FA MA M A M A M A M A M A MA FA FA FA FA FA FA FA FA MA M A MA M A M A M A M A MA FA FA FA FA FA FA FA MA MA M A MA MA MA MA MA FA FA FA FA FA FA FA M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A FD 2 F s M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A F A FM 3 MA MA MA MA MA MA MA MA MA MA FA FA FA FA MA M A M A M A MA MA MA FA FA FA FA FA FA FA FA FA FA NOT PREGNANT MA MA MA MA MA MA MA MA MA MA MA MA FA FA FA FA MORIBUND SACRIFICED ON DAY 16 OF STUDY M A M A M A M A M A M A M A M A MD 3 F A F A F A F A F A F A F A M A M A M A M A MD 2 F A F A F A F A F A F A F A F A F A F A M A M A M A MA M A FA FA FA FA FA FA FA FA FA NOT PREGNANT MA M A M A M A M A FA FA FA FA FA FS MA MA MA MA MA MA MA MA FA FA FA FA FA FA M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A FD 5 M A M A M A M A MA MA MA FA FA FA FA FA MA M A M A FA FA FA FA FA FA FA FA FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. aseooo 418-018:PAGE B-278 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-G295.25) TABLE 38 (PAGE 6): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP tf 1 2 3 4 5 6 7 S 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER # MATERNAL DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 10973 10977 10979 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 MM 3 MM 3 MD 2 M s FM 3 FM 3 FM 3 FD 2 MD 3 MM 3 MM 3 MM 3 MM 3 MD 2 MD 2 FM 4 FM 3 FM 3 FM 3 FD 2 FD 2 M A M A M A M A MM 2 MM 2 MD 2 MD 1 MD 1 F A FM 2 FD 2 FD 2 FD 2 FD 1 M A M A M A M A M A M A M A MD 4 MD 1 F A F A F A F A FD i M A M A M A M A M A M A M A M A M A MD 3 MM 3 MM 3 F A F A F A MD 3 MD 3 MD 3 MD 3 MD 3 MD 3 MD 2 FD 3 FD 3 FD 3 FD 3 FD 3 FD 3 FD 3 FD 2 FD 2 MM 3 MD 2 MD 2 MD 2 MD 2 MD 2 FM 3 FM 3 FD 2 FD 2 MM 3 MM 3 MM 3 MD 2 FD 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 M A M A MD 1 MD 1 F A F A F A F A FM 3 FM 3 FD 1 FD 1 FD 1 M A M A M A M A M A M A M A M A F A F A F A F A FD 1 M A MA M A M A FA FA FA FA FA FA FA FA MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MM 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FM 2 M A M A M A M A M A M A MD i F A F A F A F A F A F A MM 3 MD 2 MD 2 MM 2 MM 2 MM 2 MM 2 MM 2 MM 2 F A F A FM 3 FD 2 FD 2 FM 2 FM 2 FD 1 M A M A M A M A M A M A MM 3 MD 2 MM 2 F A F A F A F A FM 3 FM 3 FD 1 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD i FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 M A M A MD 2 F A F A F A F A FD 4 FD 4 FD 4 FD 3 FD 2 M A M A M A F A F A F A F A F A F A F A F A F A F A FD 2 M A MD 4 F A F A F A F A F A F A F A F A F A F A F A FD 1 FD 1 MD 5 MM 3 MM 3 MD 2 MM 2 MM 2 MD i M s FD 5 FM 3 FD 2 FD 2 FD 2 FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A - A L I V E , S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-279 000353 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 38 (PAGE 7) : PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP tt 1 2 3 4s e 7e 9 10 11 12 13 14 15 1G 17 18 19 20 21 22 23 RAT/ LITTER 0 MATERNAL DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10986 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 MD 3 MD 3 MD 3 MD 3 MD 3 MM 3 MM 3 MD 2 FD 3 FD 3 FD 3 FM 3 FD 2 FD 2 FD 2 MM 3 MD 2 MD 2 MM 2 MM 2 MM 2 MM 2 FD 2 FD 2 FM 2 FM 2 FM 2 FM 2 FM 2 FM 2 M A M A M A MD 4 MD 2 MM 2 MM 2 F A F A F A FM 3 FD 2 FM 2 FM 2 FM 2 MM 2 MD 2 MD 2 MD 2 MD 2 FM 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 M A M A M A M A M A M A M A M A M A M A M A MM 2 F A F A F A F A NOT PREGNANT MM 3 MD 2 MD 2 MD 2 MD 2 MM 2 MM 2 M s FM 3 FM 3 FM 3 FD 3 FD 2 FD 2 MM 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 F s MD 5 MD 3 MD 2 MD 1 MD i MD 1 MD 1 MD 1 MD 1 FD 1 FD 1 FD i FD i MM 3 MM 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 3 FM 3 FM 3 FD 2 FD 2 FD 1 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MM 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FM 2 M A M A MD 3 MM 3 MM 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 F A F A F A FD 2 M A MD 3 MM 2 MM 2 MD 1 F A F A F A F A F A FM 2 FM 2 FM 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 5 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 MU MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MM 2 FD 2 FD 2 FD 2 FM 2 M A M A M A MM 2 MD i F A F A FM 3 FD 2 FD 1 MM 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 3 FD 2 FD 2 FD 2 MD 2 MD 2 MM 2 MM 2 MD i MD i MD i MD i FM 2 FD 1 FD 1 FD 1 FD 1 FD 1 FD i FD 1 FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A - A L I V E , S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-280 000354 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 8): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP tt 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER # MATERNAL DOSAGE GROUP B 0.4 MG/KG PFOS 10999 11000 11001 11002 11003 11004 11005 11006 1100-7 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 M A M A MA M A MA M A MA M A M A M A FA FA FA FA FS M A M A M A M A FA FA FA FA FA FA FA FA FA FA MA M A MA MA MA MA MA MA MA MA FA FA FA FA FA MA M A M A M A MA M A M A FA FA FA FA FA FA FA M A M A M A M A M A M A M A MD 1 MD 1 M S F A F A F A F A F A F S M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A FD 3 MA M A M A M A M A M s FA FA FA FA FA FA FA FA M A MA M A M A MA FA FA FA FA FA FA FA FA FA FA FA FA FS M A M A MA M A M A M A FA FA FA FA FA FA FA FA FA FA FA MA MA MA FA FA FA FA FA FA FA FA FA FA Fs MA MA MA MA MA MA MA FA FA FA FA FA FA FA Fs MA MA M A M A MA MA FA FA FA FA FA FA FA FA FA FA MA MA MA MA MA M A FA FA FA FA M A M A M A M A M A MD 4 F A F A F A F A F A F A F A FD 4 FD 2 F S MA M A M A MA M A M A MA MA FA FA FA FA FA M A M A M A MA M A MA M A FA FA FA FA FA FA FA FA FA MA MA MA M A FA FA FA FA FA Fs MA M A MA M A MA M A MA MA MA FA FA FA FA FA MA MA M A M A M A MA MA MA FA FA FA FA FA FA FA FA FA M A M A M A M A M A M A MD 1 F A F A F A F A F A F A F A F A F A F A FA FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-28 000355 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 9): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP ft 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ I,ITTER tt MATERNAL DOSAGE GROUP 9 0.8 MG/KG PFOS 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA NOT PREGNANT MA MA MA MA MA MA NOT PREGNANT MA MA MA NOT PREGNANT MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA M A MD 4 MM 2 MA MA MA MA MA MA MA FA MA MA MA MA MA MA MA MA FA MA MA MA MA MA MA MA MA FA MA MA MA MA MA FA MA MA MA MA MA MA MA MA MA FA M A MM 2 M A M A M A MD 1 FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA MA FA MA MA MA MA MA FA MA MA FA MA MA MA MA MA FA FA MA FA FA FA FA FA FA FA FA M A M A MM 3 MA FA FA FA FA FA M A MD 2 F A F A F A FD 4 FA FA FA FA FA F A F A F A F A F A FM 3 FA FA FA FA FA F A F A F A F A FD 2 F A F A F A F A FD 2 F A FD 5 FD 4 FD 3 FD 2 FS FA FA FA FA FA FA FA FS FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA FA F A FD 2 FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A - A L I V E , S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-282 000356 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 10) : PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 1 2 3 4 5 6 7 8 9 10 ii 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER # MATERNAL DOSAGE GROUP 10 1 MG/KG PFOS 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 11628 11629 M A M A M A M A M A M A MD 5 F A F A F A F A F A F A F A M A MA MA MA M A MA MA MA FA FA FA FA FA FA FA M A M A MA MA M A FA FA FA FA FA FA FA FA FA FA FA FA NOT PREGNANT M A M A M A M A M A M A F A F A F A F A F A F A F A FM 2 FM 2 FM 2 M A M A M A M A M A M A MD 2 F A F A MA MA MA MA MA FA FA FA FA FA FA FA FA FA FA FA FA M A M A M A M A M A M A MD 4 F A F A F A F A F A F A M A MA M A M A M A MA M A MA M A MA FA FA FA FA FA FA M A M A M A M A M A M A M A M A MM 2 MD 1 F A F A F A F A F A F A F A M A M A M A M A M A M A M A F A F A F A F A F A F A F A FM 2 M A M A M A M A M A FA FA FA FA FA FA FA FA FA FA M A M A M A M A M A M A M A M A M A F A F A F A F A FD 2 M A M A M A M A M A M A M A M A MM 2 MD 1 F A F A F A F A FM 5 M A M A MA M S FA FA FA FA FA FA FA FA FA FA FA F S M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A FM 5 M A M A MA M A FA FA FA FA FA FA FA FA FA FA MD 3 MD 3 MM 3 MM 3 MM 3 MD 2 MD 2 MD 2 MM 2 MM 2 FD 3 FD 2 FD 2 FM 2 FD 1 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A F A FM 2 M A M A M A M A M A M A F A F A F A F A F A FD 1 FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A - A L I V E , S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-283 000357 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 11): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 1 18 19 20 21 22 23 RAT/ LITTER 1* MATERNAL DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 M A M A M A M A M A M A F A F A F A F A F A F A F A F A F A FD i NOT PREGNANT M A M A M A M A M A M A M A M A MM 4 F A F A F A F A M A M A M A M A M A MM 4 MD 3 MM 3 MM 3 F A F A FM 4 FM 3 M A M A M A M A M s F A F A F A F A F A F A F A F A FM 4 FM 2 M A MD 5 MD 3 MD 2 F A F A F A F A F A F A F A FD 4 FD 3 M A M A M A M A MM 4 MM 3 F A F A F A F A FD 1 M A M A M A M A M A M A M A M A M A M A MD 4 MD 4 F A F A F A F A F A M A M A M A MD 1 MD i F A F A F A F A F A F A F A F A F A F A FD i M A M A M A M A MD 5 MD 4 MM 3 MD 3 MD 2 F A F A F A F A F A F A FM 3 FD 3 FD 3 FD 2 MA MA MA FA FA FA FA NOT PREGNANT M A M A M A MD 2 MD 2 F A F A FD 2 FD 2 MA M A M A M A MA MA M A M A MA FA FA FA FA FA FA FA M A M A MA M A M A M A FA FA FA FA FA FA FA M A M A M A M A M A MD 3 MD 2 MD 2 MD 2 MD 2 F A F A F A FM 4 MA MA MA MA M A MA MA MA MA FA FA FA FA FA FA FA M A M A M A M A M A M A M A F A F A F A F A F A F A F A FD 1 M A M A M A M A MD 3 F A F A F A F A F A F A F A F A F A M A M A M A M A M A M A M A MM 2 F A F A F A F A F A F A F A FD 2 FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A - A L I V E , S-STILI.BORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-284 000358 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 12): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER It MATERNAL DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11041 11042 11043 11048 11049 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 NOT PREGNANT MD 3 MD 3 MD 3 MM 3 MM 3 MM 3 F A F A F A F A F A FD 4 FD 4 FD 3 FD 3 FD 3 FD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MM 2 FD 2 FD 2 FD 2 FD 2 FD 2 FM 2 FM 2 FD 1 FD 1 NOT PREGNANT MD 2 MD 2 MD 2 MM 2 MM 2 MM 2 MM 2 FD 2 FD 2 FM 2 FM 2 FM 2 FM 2 FM 2 FD 1 M A M A MM 2 F A FM 2 FD 1 M A M A M A M A M A M A M A MM 4 F A F A F A F A F A F A F A F A M A M A M A M A MD 2 MD 1 F A F A F A F A F A F A FM 2 FM 2 FD 1 M A M A M A MA MA MA MA MA FA FA FA FA FA FA FA FA MD 5 MD 2 MD 2 MD 2 F A F A FD 4 FM 3 FD 3 FM 2 FD 2 FD 2 M A M A M A M A M A M A MD 5 MM 2 F A F A F A F A FM 2 FD 1 MM 2 MM 2 MM 2 MD 2 MD 2 MD 2 M S M S M S FM 2 FD 2 FD 2 FD 2 F S M A M A M A F A F A F A F A F A F A F A FM 2 MM 2 MM 2 MM 2 MM 2 F A FM 3 FD 2 FD 2 FM 2 FM 2 FM 2 M A M A M A M A MD 2 MD 1 F A F A F A F A F A F A FD 1 MM 5 MM 5 MM 3 MD 2 MD 2 MD 2 MM 2 MM 2 FM 3 FD 2 FD 2 FD 2 FD 2 FM 2 FM 2 FM 2 FD X M A M A M A M A M A MD 2 F A F A F A F A F A F A F A M A M A M A M A MD 4 F A F A FD 2 FD 1 FD 1 FD 1 NOT PREGNANT M A M A M A M A M A M A M A MD 2 M A F A F A F A F A FD 3 FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-285 000359 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 38 (PAGE 13) : PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER ft MATERNAL DOSAGE GROUP 13 2 MG/KG PFOS 11055 11057 11059 11061 11064 11065 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 11664 11665 11666 11667 MM 4 MM 4 MM 2 MD 2 MD 2 FM 3 FM 3 FM 2 FM 2 FM 2 FD 2 FD 2 FD 2 FD 1 MM 2 MM 2 MD 2 MD 2 FM 2 FM 2 FM 2 FD 2 FD 2 FD 1 M A M A M A M A M A M A M A M A M A M A M A F A F A FM 2 FM 2 MM 3 MM 3 MD 2 MD 2 MD 2 MD 2 MD 2 FM 3 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MM 3 FD 2 FD 2 FD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FM 2 M A M A M A M A MD 3 F A F A F A F A FD 2 FM 2 FM 2 FM 2 FM 2 M A M A M A M A F A F A F A F A F A F A F A FD 4 FM 2 FM 2 M A M A M A M A MD i MD 1 F A F A F A F A F A F A MD 3 MM 2 MD 2 MD i MD 1 MD 1 MD 1 MD 1 FM 2 FD 2 FD 1 FD 1 FD 1 MD 1 MD 1 MD i MD 1 MD 1 MD 1 MD 1 MD 1 FM 2 FM 2 FD i FD 1 FD 1 FD 1 MD 1 MD i FD i FD 1 FD i FD 1 MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 3 FM 3 FD 2 FD 2 FD 2 MD 2 MD 1 MD i MD 1 MD i MD i FD 2 FD 2 FD i FD 1 FD i FD 1 FD 1 FD 1 FD 1 MD 3 MD 3 MD 2 MD 2 MD 2 MM 2 MM 2 MM 2 MM 2 F A FM 2 FM 2 FM 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 1 FD 2 FD 2 FD 2 FD 2 FD 2 FM 2 FD 1 MD 2 MD 2 MD 2 MD 2 MD 2 MM 2 MD 1 FD 2 FD 2 FD 2 FD 1 MM 5 MM 3 MD 2 MM 2 MM 2 MM 2 FM 5 FD 3 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FM 2 FM 2 NOT PREGNANT MM 2 MM 2 MD 1 MD 1 MD 1 MD 1 MD 1 M S FM 2 FM 2 FD 1 FD 1 FD 1 FD i FD 1 FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER'1 INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. senno 418-018-.PAGE B-286 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 39 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY (S) POSTPARTUM OBSERVATIONS a 1 VEHICLE CONTROL 10906 4- 5 1/15 PUPS: TIP OF TAIL BLACK.b 11504 2 1/15 PUPS: COLD T O TOUCH. 2 MEVALONIC ACID CONTROL - - - 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 3 1/11 PUPS: NOT NURSING. 10941 3 3/ 4 PUPS: COLD TO TOUCH. NOT NURSING. 11530 3 1/10 PUPS : DECREASED MOTOR ACTIVITY. 11532 2 1/11 PUPS : NO MILK IN STOMACH. 11542 1 15/15 PUPS : NOT NURSING. 4 2 MG/KG PFOS + MEVALONIC ACID 10944 4 1/ 1 PUP: NOT NURSING. 11547 5 1/ 3 PUPS: NOT NURSING. 11549 1 1/14 PUPS: DISTAL PORTION OF TAIL MISSING. 11550 2 6/12 PUPS: COLD TO TOUCH. 11552 5 2/ 2 PUPS: COLD TO TOUCH, 11553 1 17/17 PUPS: NOT NURSING. 2 2/17 PUPS : COLD TO TOUCH. 11554 1 15/17 PUPS: NOT NURSING. 11555 1 1/14 PUPS: NOT NURSING. 2 2/14 PUPS: COLD TO TOUCH. a. Tabulation restricted to adverse observations; all other pups appeared normal. b. Observation confirmed at necropsy. laeaeo 418-018:PAGE B-287 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 9 (PAGE 2) : CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY (S) POSTPARTUM OBSERVATIONS a 5 CHOLESTEROL CONTROL 10957 3- 5 i/ii PUPS: TIP OF TAIL BLACK.b 10967 4- 5 1/15 PUPS: RIGHT EYE, E X O P H T H A L M O S .b 11559 4- 5 1/18 PUPS: RIGHT EYE, EXOPHTHALMOS. 11563 4 1/15 PUPS: SWOLLEN 56,57. COLD TO TOUCH. 11567 4- 5 1/10 PUPS: TIP OF TAIL MISSING.b 6 1.6 MG/KG PFOS + CHOLESTEROL 10984 1 1/16 PUPS: COLD TO TOUCH. NOT NURSING. 2 13/16 PUPS: COLD TO TOUCH. 11572 2 3/10 PUPS: COLD TO TOUCH. 11573 2 4/13 PUPS: NOT NURSING. 11574 2 8/ 8 PUPS: COLD TO TOUCH. 11577 1 2/16 PUPS: DECREASED MOTOR ACTIVITY. 2 1/16 PUPS: COLD TO TOUCH. NOT NURSING. 11578 1 2- 5 1/13 PUPS: DISTAL PORTION OF TAIL MISSING. 1/12 PUPS: DISTAL PORTION OF TAIL MISSING.b 11580 2 13/15 PUPS: COLD TO TOUCH. 3 10/13 PUPS: COLD TO TOUCH. a. Tabulation restricted to adverse observations; all other pups appeared normal. b. Observation confirmed at necropsy. 418-018:PAGE B-288 000362 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER; T-6295.25) TABLE 3 9 (PAGE 3): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY (S) POSTPARTUM OBSERVATIONS a 7 2 MG/KG PFOS + CHOLESTEROL 10986 2 11/15 PUPS : COLD TO TOUCH. DECREASED MOTOR ACTIVITY. 10994 1 2/15 PUPS: NOT NESTING. NOT NURSING. DECREASED MOTOR ACTIVITY. 10995 5 6/ 6 PUPS: COLD TO TOUCH. 10997 1 1/16 PUPS: COLD TO TOUCH. NOT NESTING. 5 1/15 PUPS: COLD TO TOUCH. NOT NESTING. NOT NURSING. 11587 2 1/13 PUPS: NOT NURSING. 11588 1 13/14 PUPS: NOT NURSING. 11589 4 1/ 1 PUP: NOT NURSING. 11590 i 15/16 PUPS: NOT NURSING. 2 5/15 PUPS: COLD TO TOUCH. 11592 1 15/15 PUPS: NOT NURSING. 11593 2 3 4- 5 1/12 PUPS: TAIL MISSING. 1/ 7 P U P S : TAIL MISSING. 1/ 6 PUPS: TAIL MISSING.b 11596 2 1/14 PUPS: COLD TO TOUCH. NOT NURSING. 11598 2 2/15 P U P S : COLD TO TOUCH. 8 0.4 MG/KG PFOS 11000 1 1/14 PUPS: DECREASED MOTOR ACTIVITY. GASPING. NOT NURSING. NOT NESTING. 11603 3 1/14 PUPS: DECREASED MOTOR ACTIVITY. LABORED BREATHING. COLD TO TOUCH. a. Tabulation restricted to adverse observations; all other pups appeared normal. b. Observation confirmed at necropsy. 418-018:PAGE B-289 000363 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 3 9 (PAGE 4): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY (S) POSTPARTUM OBSERVATIONS a 9 0.8 MG/KG PFOS 11009 4- 5 1/13 PUPS: TIP OF TAIL BLACK.b 11015 1 16/16 PUPS: COLD TO TOUCH. 11617 i 1/17 PUPS: NOT NURSING. 10 1 MG/KG PFOS 11620 2 3- 5 1/15 PUPS: TIP OF TAIL MISSING. 1/14 PUPS: TIP OF TAIL MISSING.b 11624 1 14/16 PUPS: COLD TO TOUCH. 11628 1 1/17 PUPS: DECREASED MOTOR ACTIVITY. 11 1.2 MG/KG PFOS 11035 1 10/11 PUPS: COLD TO TOUCH. 2 10/10 PUPS: COLD TO TOUCH. 11038 3 12/17 PUPS: COLD TO TOUCH. 11630 1- 5 1/ 7 PUPS: TAIL MISSING FROM B A S E .b 11633 2 16/16 PUPS: COLD TO TOUCH. 11635 3 1/10 PUPS: NOT NURSING. 12 1.6 MG/KG PFOS 11041 5 1/ 5 PUPS: DECREASED MOTOR ACTIVITY. COLD TO TOUCH. 11643 4 1/ 4 PUPS: COLD TO TOUCH. NOT.NURSING. 11647 5 1/ 1 PUP: COLD TO TOUCH. NOT NURSING. a. Tabulation restricted to adverse observations; all other pups appeared normal. b. Observation confirmed at necropsy. 418-018:PAGE B-290 tseooo PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 39 (PAGE 5): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY (S) POSTPARTUM OBSERVATIONS a 13 2 MG/KG PFOS 11055 3 1/ 4 PUPS: COLD TO TOUCH. NOT N U R S I N G . 1/ 4 PUPS: NOT NURSING. 11065 2 2/14 PUPS: COLD TO TOUCH. 11662 5 1/ 1 PUP: COLD TO TOUCH. NOT NURSING. 11665 4 2/ 2 PUPS: NOT NURSING. a. Tabulation restricted to adverse observations; all other pups appeared normal. 418-018:PAGE B-2 000365 vo PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 1 VEHICLE CONTROL 10906 5 ALL PUPS APPEARED NORMAL. 10909 i 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 10911 4 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 10913 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 10914 5 ALL PUPS APPEARED NORMAL. 11501 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11502 1 1 PUP: STILLBORN. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11503 5 ALL PUPS APPEARED NORMAL. 11504 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION . 5 ALL PUPS APPEARED NORMAL. 11505 11506 11507 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11508 1 2 PUPS: STILLBORN. TONGUE PROTRUDED. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11509 11510 11511 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 99E000 418-018:PAGE B-292 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 1 VEHICLE CONTROL (CONT.) 11512 1 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11513 11514 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 2 MEVALONIC ACID CONTROL 10921 10922 10923 10924 10927 11515 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11516 I 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 A LL PUPS APPEARED NORMAL. 11517 11518 5 A L L PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11519 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11520 11521 11523 11524 11525 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED N O R M A L .. 11526 1 1 PUP: STILLBORN, NO ORAL OPENING, NO EYES, EARS DISPLACED. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11527 11528 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 000 418-018:PAGE B-293 PROTOCOL 418-016: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 2 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 10938 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 10941 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 10942 5 ALL PUPS APPEARED NORMAL. 11529 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11530 1 2 PUPS: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11532 1 2 PUPS: STILLBORN. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 6 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018-.PAGE B-294 000368 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 1.6 MG/KG PFOS + MEVALONIC ACID (CONT.) 11533 1 8 PUPS: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11534 1 1 PUP: STILLBORN. A LL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 6 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11535 2 13 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 11536 11537 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11538 1 4 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11539 2 6 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11540 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 5 PUPS: FOUND DEAD. AUTOLYSIS. PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11541 2 10 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-295 000369 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 5): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 3 1.6 MG/KG PFOS + MEVALONIC ACID (CONT.) . 11542 2 7 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. A LL TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11543 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 4 2 MG/KG PFOS + MEVALONIC ACID 10943 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 8 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 i PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 10944 2 3 PUPS : FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 10948 1 10 PUPS: FOUND DEAD. NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 10952 2 14 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 10955 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 7 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 6 PUPS: FOUND DEAD. NO MILK IN STOMACH. A UTO LYSIS PRECLUDED FURTHER EVALUATION. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-296 000370 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 6) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 4 2 MG/KG PFOS + MEVALONIC ACID (CONT.) 10956 1 I PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 13 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11544 2 14 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11545 1 3 PUPS: FOUND DEAD. NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11546 1 ii PUPS: FOUND DEAD. N O MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11547 1 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 2 3 PUPS : FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 PUP APPEARED NORMAL, 11549 1 5 PUPS: FOUND DEAD. NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 4 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11550 i 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 4 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-297 000371 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 7) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 4 2 MG/KG PFOS + MEVALONIC ACID (CONT.) 11551 1 2 PUPS: FOUND DEAD. AL L TISSUES APPEARED NORMAL. 7 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11552 1 7 PUPS : FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 i PUP: FOUND DEAD. NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11553 2 15 PUPS: FOUND DEAD. AL L TISSUES APPEARED NORMAL. 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11554 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 2 3 PUPS: FOUND DEAD. AUTO LYSIS PRECLUDED FURTHER EVALUATION. 10 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11555 2 12 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11556 1 16 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 11557 1 10 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018(PAGE B-298 000372 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY-NUMBER: T-6295.2S) TABLE 40 (PAGE 8): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 5 CHOLESTEROL CONTROL 10957 10966 10967 10968 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 10969 1 1 PUP: STILLBORN, ALL TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 10970 11558 11559 11561 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11563 3 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11564 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11565 5 ALL PUPS APPEARED NORMAL. 11567 1 1 PUP: STILLBORN, ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11568 5 ALL PUPS APPEARED NORMAL. 11569 5 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. ALL OTHER PUPS APPEARED NORMAL. 11570 11571 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018(PAGE B-299 000373 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 9): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 i 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 10973 2 4 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. CANNIBALIZATION PRECLUDED FURTHER EVALUATION. 10977 1 3 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 4 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 10979 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 10983 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 10984 2 3 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 12 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 11572 2 6 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-300 000374 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 10): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a E 1.6 MG/KG PFOS + CHOLESTEROL (CONT.) 11573 2 9 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11574 1 5 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL 5 ALL PUPS APPEARED NORMAL. 11575 i 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11576 5 ALL PUPS APPEARED NORMAL. 11577 2 13 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11578 1 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11579 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 4 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11580 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-301 000375 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 11) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 6 1.6 MG/KG PFOS + CHOLESTEROL (CONT.) 11581 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 12 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11582 2 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 3 PUPS: FOUND DEAD. A LL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11583 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11584 1 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11585 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 4 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-302 000376 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 12): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 7 2 MG/KG PFOS + CHOLESTEROL 10986 2 4 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 e PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 10994 2 4 PUPS: FOUND DEAD. NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 10995 2 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 4 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 10996 2 11 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 10997 5 ALL PUPS APPEARED NORMAL. 11586 1 1 PUP: STIL L B O R N . ALL TISSUES APPEARED NORMAL. 2 6 PUPS: FOUND DEAD. ALL TISSUES A PPEARED NORMAL. 3 1 PUP: FOUND DEAD. ALL TISSUES A PPEARED NORMAL. 11587 2 12 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11588 1 i PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 13 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11589 1 9 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION, 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION, 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-303 000377 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 13): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 7 2 MG/KG PFOS + CHOLESTEROL (CONT.) 11590 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 8 PUPS: FOUND DEAD. AL L TISSUES APPEARED NORMAL. 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11591 1 i PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 14 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11592 2 7 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11593 i 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. N O MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11594 2 15 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11596 2 11 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION, 3 1 PUP: FOUND DEAD, AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11597 1 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION, 2 1 PUP: FOUND DEAD, AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-304 000378 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 14): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 2 MG/KG PFOS + CHOLESTEROL (C O N T .) 11598 2 13 PUPS : FOUND DEAD. ALL TISSUES APPEARED NORMAL 8 0.4 MG/KG PFOS 11599 10999 1 11 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 2 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11000 11001 11002 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11003 1 2 PUPS: STILLBORN. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11004 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11005 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11006 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11007 5 ALL PUPS APPEARED NORMAL. 11008 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. a . Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-305 000379 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 15): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 8 0.4 MG/KG PFOS (CONT.) 11600 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11601 11602 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11603 1 1 PUP: STILLBORN. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 2 1 PUP: FOUND DEAD. A LL TISSUES APPEARED NORMAL. 4 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11604 5 ALL PUPS APPEARED NORMAL. 11605 5 1 PUP: LIVER, MEDIAN LOBE, TAN AREA (4.0 CM IN DIAMETER). ALL OTHER PUPS APPEARED NORMAL.b 11606 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11607 11608 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11609 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 9 0.8 MG/KG PFOS 11009 11010 11011 11012 5 ALL PUPS APPEARED NORMAL 5 ALL PUPS APPEARED NORMAL 5 ALL PUPS APPEARED NORMAL 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. b. Necropsy observations were not recorded for ten of the sixteen pups. 000 418-018(PAGE B-306 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 16): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 9 0.8 MG/KG PFOS (CONT.) 11013 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11015 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. ALL OTHER PUPS APPEARED NORMAL. 11016 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11018 11611 11612 11613 11614 11615 11616 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11617 2 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11618 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11619 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-307 000381 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 17): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 10 1 MG/KG PFOS 11019 5 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. ALL OTHER PUPS APPEARED NORMAL. 11020 11021 11023 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11024 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11025 5 ALL PUPS APPEARED NORMAL. 11026 4 1 PUP: FOUND DEAD. AL L TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11027 5 ALL PUPS APPEARED NORMAL. 11028 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11620 11621 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11622 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11623 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11624 1 2 PUPS: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-308 000382 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-629S.25) TABLE 40 (PAGE 18): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 10 1 MG/KG PFOS (CONT.) 11625 11626 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11627 1 1 PUP: FOUND DEAD. NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 4 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 3 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11628 5 ALL PUPS APPEARED NORMAL. 11629 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11 1.2 MG/KG PFOS 11029 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11031 5 ALL PUPS APPEARED NORMAL. 11032 3 1 PUP: FOUND DEAD. NO M ILK IN S T O M A C H . AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11033 1 1 PUP: STILLBORN. ALL TISSUES APPEARED N O R M A L . 5 ALL PUPS APPEARED NORMAL. 11034 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 4 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. ALL OTHER PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-309 000383 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 19): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 11 1.2 MG/KG PFOS (CONT.) 11035 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11036 4 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11037 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11038 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. AL L TISSUES APPEARED NORMAL. 5 1 PUP: FOUND DEAD. NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. ALL OTHER PUPS APPEARED NORMAL. 11630 5 ALL PUPS APPEARED NORMAL. 11632 2 4 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11633 11634 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11635 2 4 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-310 000384 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T - 6 2 9 5 .25) TABLE 40 (PAGE 20): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 11 1.2 MG/KG PFOS (CONT.) 11636 5 ALL PUPS APPEARED NORMAL. 11637 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11638 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11639 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 12 1.6 MG/KG PFOS 11041 2 1 PUP: FOUND DEAD, NO M ILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 3 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11042 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 11 PUPS; FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11048 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 3 PUPS: FOUND DEAD. CANNIBALIZATION PRECLUDED FURTHER EVALUATION. 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11049 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL, 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-311 000385 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 21): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 12 1.6 MG/KG PFOS (CONT.) 11640 5 ALL PUPS APPEARED NORMAL. 11641 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11642 5 ALL PUPS APPEARED NORMAL. 11643 2 5 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL OTHER PUPS APPEARED NORMAL. 11644 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL OTHER PUPS APPEARED NORMAL. 11645 1 4 PUPS: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. CANNIBALIZATION PRECLUDED FURTHER EVALUATION. 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11646 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-312 000386 418-018(PAGE B000387 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 22): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 12 1.6 MG/KG PFOS (CONT.) 11647 2 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 PUP APPEARED NORMAL. 11648 1 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11649 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 7 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11650 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11651 1 3 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 4 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11653 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed' at necropsy. Oj UJ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) TABLE 40 (PAGE 23): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 13 2 MG/KG PFOS 11055 1 1 PUP ; FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 5 PUPS: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11057 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 4 PUPS : FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11059 5 ALL PUPS APPEARED NORMAL. 11061 2 12 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11064 2 7 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 2 PUPS : FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11065 2 11 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11654 2 i PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 i PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11655 4 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11656 1 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-3I4 000388 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS MEVALONIC AC ID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 24): NECROPSY OBSERVATIONS - INDIVIDUAL DATA FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 13 2 MG/KG PFOS (CONT.) 11657 1 3 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 11658 1 7 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11659 i 6 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 11660 2 10 PUPS : FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11661 1 11 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. i PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 3 PUPS : FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11662 2 3 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 PUP APPEARED NORMAL. 11663 I 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 13 PUPS : FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11664 i 2 PUPS: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 8 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 6$eooo 418-018:PAGE B-315 PROTOCOL 418-0X8: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 40 (PAGE 25): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 13 2 MG/KG PFOS (CONT.) 11665 2 6 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11667 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 9 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 39) for external clinical observations confirmed at necropsy. 418-018:PAGE B-316 0003# APPENDIX C PROTOCOL AND AMENDMENTS 000391 418-018:PAGE C-l O Primedica Argus Research Laboratories. Inc. 905 Sheehy Drive, Building A Horsham. PA 19044 Telephone: (215) 445-8710 Telefax: (215) 445-8587 PROTOCOL 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 STUDY TITLE: One Generation Reproduction Study of PFOS - Mevalonic Acid/Cholesterol Challenge and NOEL Investigation in Rats PURPOSE: The purposes of this study are to determine whether or not co-administration of mevalonic acid or cholesterol can prevent or antagonize the adverse maternal and fetal effects (reduced maternal body weight gain, increased percent stillborn, decreased birth weight and decreased pup survival) observed in a previous study (Argus Research Protocol 418-008, 3M study number T-6295.9) and to better define the no observed effect level (NOEL). TESTING FACILITY: Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Telephone: (215)443-8710 Telefax: (215)443-8587 STUDY DIRECTOR: Raymond G. York, Ph.D., DABT Associate Director of Research raymond.york @primedica.com SPONSOR: 3M Corporate Toxicology 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 STUDY MONITOR: Deanna J. Luebker, M.S. 3M Corporate Toxicology 3M Medical Department Telephone: (651)737-1374 Telefax: (651)733-1773 ALTERNATE STUDY MONITOR: John Butenhoff, Ph.D., DABT, CIH 3M Corporate Toxicology 3M Medical Department Telephone: (651)737-1962 Telefax: (651)733-1773 000392 418-018:PAGE C- REGULATORY CITATIONS: Protocol 418-018 Page 2 Study Design as Modification of: U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Federal Register, September 22, 1994, Vol. 59, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. REGULATORY COMPLIANCE: This study will be conducted in compliance with the Good Laboratory Practice (GLP) regulations cited above. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. The Testing Facility's Quality Assurance Unit (QAU) will audit the protocol, the raw data and the report, and will inspect critical phases of those portions of the study conducted at the Testing Facility in accordance with the Standard Operating Procedures of Argus Research Laboratories, Inc. Should any portion of the study be conducted by a subcontractor or by the Sponsor, the QAU for that facility will conduct critical phase inspections and audit respective results and reports for that study portion according to the SOPs of that facility. Such critical phase inspection reports and report audits will be submitted by that facility to the Study Director. The dates of the inspections and report submissions will be incorporated into a QAU Statement generated by that facility and provided to the Testing Facility for inclusion in the final report. The final report will include a compliance statement signed by the Study Director that the report accurately reflects the raw data obtained during the performance of the study and that all applicable GLP regulations were followed in the conduct of the study. Should significant deviations from GLP regulations occur, each will be described in detail, together with how the deviation might affect the quality or integrity of the study. 000393 418-018:PAGE C-3 STUDY SCHEDULE: Protocol 418-018 Page 3 See ATTACHMENT 1 to the protocol. TEST ARTICLE AND VEHICLE: Identification: Test Article: Name: Physical Description: Lot/Batch Number: Specific Gravity: Purity: Expiration Date: Perfluorooctane Sulfonic Acid Potassium Salt (PFOS). Light-colored powder. 217 0.6 98.9% NA Information on the identity, composition, strength and purity of the test article is on file with the Sponsor. Control Articles: Name: Physical Description: Lot Number: Specific Gravity: Purity: Expiration Date: Mevalonic Acid. White with dark yellow cast, semisolid. 070K2603 NA Approximately 97% August 2004 Name: Physical Description: Lot Number: Specific Gravity: Purity: Expiration Date: Cholesterol. White powder 119H0218 NA 95% July 2004 Information on the identity, composition, strength and purity of the control articles will be maintained in the raw data. Vehicles: 0.5% Tween 80 in reverse osmosis membrane processed deionized water (RODI H2O). Supplier and lot identification of Tween 80 to be documented in the raw data. Reverse Osmosis Membrane Processed Deionized Water (RODI H2O). 000394 418-018:PAGE C-4 Protocol 418-018 Page 4 Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the vehicles that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this protocol will be conducted. Safety Precautions: Gloves, dust-mist/HEPA*filtered mask, appropriate eye protection and a uniform/lab coat are to be worn during formulation preparation and dosage administration. The Material Safety Data Sheet (MSDS) is attached to the protocol (ATTACHMENT 2). Storage: Bulk Test Article: Control Article Components: Vehicle Components: Prepared Vehicle (0.5% Tween 80): Prepared Test Article Formulations: Room temperature. Room temperature. Room temperature. Room temperature. Frozen (-20C). All test article shipments to the Testing Facility should be addressed to the attention of Julian Gulbinski, Manager of Formulations, at the previously cited address and telephone number. Shipments should include information concerning storage conditions and shipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment. FORMULATION: Frequency of Preparation: Formulations of PFOS (suspensions) will be prepared weekly at the Testing Facility. The vehicle (0.5% Tween 80) will be prepared weekly. Mevalonic acid will be prepared twice daily in reverse osmosis membrane processed deionized water. Cholesterol will be prepared as a suspension once daily in 0.5% Tween 80. Detailed preparation procedures are attached to this protocol (ATTACHMENT 3) and documentation of formulation preparation will be maintained in the raw data. 000395 418-018:PAGE C-5 Adjustment for Purity: Protocol 418-018 Page 5 The test and control articles will be considered 100% pure for the purpose of dosage calculations. Testing Facility Reserve Samples: The Sponsor will reserve a sample (1 g) of each lot of the bulk test article used during the course of this study. The Testing Facility will reserve a sample (1 g or 5 mL) of each lot of the control article and vehicle components used during the course of this study. Samples will be stored under the previously cited conditions. ANALYSES: Samples additional to those described below may be taken if deemed necessary during the course of the study. Bulk Test Article Sampling: No analyses of the bulk test article will be conducted during the course of this study. Information on the stability of the bulk test article is on file with the Sponsor. The Certificate of Analysis is on file with the Sponsor and a copy will be included in the final report. Analyses of Prepared Formulations: Stability: Stability data for prepared test article formulations bracketing the range of concentrations and conditions of this study are on file with the Sponsor and will not be determined during the conduct of this study. Test article suspensions will be prepared weekly at the Testing Facility. Homogeneity Analyses: Homogeneity of the test article in prepared suspensions will be verified during the course of this study. A syringe will be used to withdraw samples (5 mL each) from the top, middle and bottom of the highest concentration on the first day of preparation. Each sample (5 mL) will be divided into two aliquots and placed into glass containers, one of 2 mL and one of 3 mL. One aliquot (2 mL) will be shipped for analysis: the other aliquot (3 mL) will be retained at the Testing Facility as a backup sample. Backup samples will be stored under the previously cited conditions and discarded at the Testing Facility upon the request of the Sponsor. 000396 418-018:PAGE C-6 Concentration Analyses: Protocol 418-018 Page 6 Concentration of the prepared test article suspensions will be verified during the course of this study. A syringe will be used to withdraw samples (5 mL each) from each concentration once during each of the following periods of the study: premating, gestation and postnatal. Each sample (5 mL each) will be divided into two aliquots and placed into glass containers, one of 2 mL and one of 3 mL. One aliquot (2 mL) will be shipped for analysis: the other aliquot (3 mL) will be retained at the Testing Facility as a backup sample. Backup samples will be stored under the previously cited conditions and discarded at the Testing Facility upon the request of the Sponsor. Shipping Instructions: Samples to be analyzed will be shipped (frozen on dry ice) to: Dave Ehresman 3M Corporate Toxicology 3M Center Building 236-0C-48 St. Paul, Minnesota 55144 Telephone: (651)733-5070 Telefax: (651)733-1773 Both the recipient and the Study Monitor will be notified in advance of sample shipment. DISPOSITION: Prepared formulations will be discarded at the Testing Facility. All remaining bulk test article will be returned to the Study Monitor at the previously cited address. TEST SYSTEM: Species/Strain and Reason for Selection: The Crl:CD(SD) IGS BR VAF/Plus rat was selected as the Test System because: 1) this strain of rat has been demonstrated to be sensitive to reproductive and developmental toxins and has been widely used throughout industry for reproductive and developmental toxicity evaluations: 2) historical data and experience exist at the Testing Facility11'31; 3) the test article is pharmacologically active in the species and strain; and 4) this strain has been used in previous reproduction studies of PFOS. GG0397 418-018:PAGE C-7 Number: Protocol 418-018 Page 7 Initial population acclimated: 360 virgin female rats (two shipments of 180 female rats each designated as Replicates 1 and 2). Population selected for study: 332 female rats (28 in each of Groups 1 to 7, 12 and 13, and 20 in each of Groups 8 to 11). Eight mated female rats in each of Groups 1 to 7, 12 and 13 (Replicate 1) will be assigned to Caesarean-sectioning on day 21 of presumed gestation; the remaining female rats will be permitted to deliver litters. Body Weight and Age: Female rats will be ordered to weigh from 175 g to 225 g each at receipt, at which time they will be expected to be at least 56 days of age. Actual body weights will be recorded the day after receipt and will be documented in the raw data. The weight ranges will be included in the final report. Sex: Female rats will be given the test article. Male rats of the same source and strain will be used only as breeders and are not considered part of the Test System. Source: Charles River Laboratories, Inc. The rats will be shipped in filtered cartons by air freight and/or truck from Charles River Laboratories, Inc., to the Testing Facility. Identification: Fo Generation: Rats are permanently identified using Monel self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats are given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats are assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study on the basis of physical appearance and body weights recorded during acclimation. F1 Generation: Pups will not be individually identified during lactation: all parameters will be evaluated in terms of the litter. 00039 418-018:PAGE C-8 ANIMAL HUSBANDRY: Protocol 418-018 Page 8 All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratory Animalsw . Housing: Fo Generation Rats/F1 Generation Litters: Fo generation rats will be individually housed in stainless steel wire-bottomed cages except during the cohabitation and postpartum periods. During cohabitation, each pair of rats will be housed in the male rat's cage. Beginning no later than day 20 of presumed gestation, Fo generation female rats assigned to natural delivery will be individually housed in nesting boxes. Each dam and delivered litter will be housed in a common nesting box during the postpartum period. Nesting Material: Bedding material (bed-o'cobs) will be supplied to female rats assigned to natural delivery. Bedding will be changed as often as necessary to keep the animals dry and clean. Analyses for possible contamination are conducted semi-annually and documented in the raw data. Room Air, Temperature and Humidity: The animal room is independently supplied with at least ten changes per hour of 100% fresh air that has been passed through 99.97% HEPA filters. Room temperature will be maintained at 64F (18C) to 79F (26CC) and monitored constantly. Room humidity will also be monitored constantly and maintained at 30% to 70%. Light: An automatically controlled 12-hour light: 12-hour dark fluorescent light cycle will be maintained. Each dark period will begin at 1900 hours EST. Diet: Rats will be given Certified Rodent Diet #5002 (PMI Nutrition International) available ad libitum from individual feeders. 000399 418-018:PAGE C-9 Water: Protocol 418-018 Page 9 Water will be available ad libitum from individual bottles attached to the cages or from an automatic watering access system. All water will be from a local source and passed through a reverse osmosis membrane before use. Chlorine will be added to the processed water as a bacteriostat; processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. Contaminants: Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the drinking water or the nesting material at levels that would interfere with the results of this study. The Sponsor is aware of a potential contaminant present at extremely low levels in the certified diet. This low-level contamination will not interfere with the results of this study. Therefore, no analyses other than those routinely performed by the feed supplier or those mentioned in this protocol will be conducted. RANDOMIZATION AND COHABITATION: Fo Generation: In order to facilitate scheduling, female rats will be received in two shipments (180 rats each) separated by two weeks. The first shipment will be designated as Replicate 1 and the second shipment will be designated as Replicate 2. Upon arrival, rats will be assigned to individual housing on the basis of computer-generated random units. After acclimation, female rats will be selected for study on the basis of physical appearance and body weights recorded during acclimation. The rats will be assigned to dosage groups based on computer-generated (weight-ordered) randomization procedures. Within each dosage group, consecutive order will be used to assign rats to cohabitation, one male rat per female rat. The cohabitation period will consist of a maximum of 14 days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ will be considered to be at day 0 of presumed gestation and assigned to individual housing. Female rats not mated within the first 7 days of cohabitation will be assigned alternate male rats that have mated and will remain in cohabitation for a maximum of seven additional days. The first eight female rats per dosage group (groups 1 to 7, 12 and 13, Replicate 1) with a confirmed date of mating will be assigned to Caesarean-sectioning on day 21 of presumed gestation. The remaining female rats (including those with no confirmed mating date) will be permitted to naturally deliver litters. 000400 418-018:PAGE C-10 F1 Generation Pups: Protocol 418-018 Page 10 Day 1 of lactation (postpartum) is defined as the day of birth and is also the first day on which all pups in a litter are individually weighed (pup body weights will be recorded after all pups in a litter are delivered and groomed by the dam). ADMINISTRATION: Route and Reason for Choice: The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered: 2) it is one of the possible routes of human exposure; and 3) it is the route used in previous reproduction studies on PFOS. Method and Frequency: Dosages will be adjusted for the most recently recorded body weight and given at approximately the same times each day. Dosages will be administered according to the table in the Dosage Levels, Concentrations and Volumes section of the protocol. The mevalonic acid dosing needle will be wiped clean before administration for each animal. Fo Generation Female Rats: Female rats will be given the test article daily beginning 42 days before cohabitation (maximum of 14 days) and continuing through day 20 of presumed gestation (rats assigned to Caesarean-sectioning), day 24 of presumed gestation (rats assigned to natural delivery that do not deliver a litter) or day 4 postpartum (rats that deliver a litter). Rats in the process of delivering will not be given test/control article; such animals may not receive any or all of their daily dosage(s) on the day of parturition. F1 Generation: F1 generation pups will not be directly given the test article, but will be possibly exposed to the test article during maternal gestation (in utero exposure) or via maternal milk during the lactation period. Rationale for Dosage Selection: Dosages were selected by the Sponsor on the basis of previous studies conducted with the test article. 000491 418-018:PAGE C-ll Dosage Levels. Concentrations and Volumes: Protocol 418-018 Page 11 Number 1 2 3 4 5 6 7 8 9 10 11 12 13 Dosage Groups Identification Vehicle Control Mevalonic Acid Control 1.6 mg/kg PFOS + Mevalonic Acid 2 mg/kg PFOS + Mevalonic Acid Cholesterol Control 1.6 mg/kg PFOS + Cholesterol 2 mg/kg PFOS + Cholesterol 0.4 mg/kg PFOS 0.8 mg/kg PFOS 1 mg/kg PFOS 1.2 mg/kg PFOS 1.6 mg/kg PFOS 2 mg/kg PFOS Number of Animals 28 28 28 28 28 28 28 20 20 20 20 28 28 Dosage Groups and Dosing Schedule 1*1dosage Dosing Schedule 2 " a dosage '/? hour 10 minutes after 1" RODI HjO 0.5% Tween 80 500 mg/kg Mevalonic Acid 0.5% Tween 80 500 mg/kg Mevalonic Acid 1.6 mg/kg PFOS 500 mg/kg Mevalonic Acid 2 mg/kg PFOS 500 mg/kg Cholesterol 0.5% Tween 80 500 mg/kg Cholesterol 1.6 mg/kg PFOS 500 mg/kg Cholesterol 2 mg/kg PFOS NA 0.4 mg/kg PFOS NA 0.8 mg/kg PFOS NA 1 mg/kg PFOS NA 1.2 mg/kg PFOS NA 1.6 mg/kg PFOS NA 2 mg/kg PFOS 3r dosage 5 hr 30 minutes after 2" RODI HjO 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid NA NA NA NA NA NA NA NA NA Compound Identification Dosage (mg/kg/day) Concentration (mg/mL) Dosage Volume imL/kg) Argus Batch Number RODI HjO 0 0 5 B-418-018-AfDay.Month.Year) 0.5% Tween 80 0 0 5 B-418-018-BfDay.Month.Year) Mevalonic Acid 1000 100 5- B-418-018-C(Day.Month. Year) Cholesterol 500 100 5 B-418-018-D(Dav.Month. Year) PFOS 0,4 0.08 5 B-418-018-EfDav.Month.Year) PFOS 0.8 0.16 5 B-418-018-F(Dav.Month.Year) PFOS 1 0 20 5 B-418-018-G(Dav.Month.Year) PFOS 1.2 0.24 5 B-418-018-HfDav.Month.Year) PFOS 1.6 0.32 5 B-418-018-l(Dav.Month. Year) PFOS 2 0.40 5 B-418-018-J(Day.Month. Year) The test and control articles will be considered 100% pure tor the purpose ot dosage calculations. ' Mevalonic acid will be dosed twice daily. 000402 418-018:PAGE C-12 TESTS. ANALYSES AND MEASUREMENTS - Fo GENERATION: Protocol 418-018 Page 12 Viability: All Periods: At least twice daily. Clinical Observations and/or General Appearance: Acclimation Period: At least once. Dosage Period: Prior to dosage administration, approximately one hour after the second dosage and at the end of the working day (approximately one hour after the third dosage). Day of Sacrifice: Once. Maternal Behavior: Days 1 and 5 postpartum. Any observed abnormal behavior will be recorded daily. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or Study Monitor. Body Weights: Acclimation Period: At least once. Dosage Period: Weekly to cohabitation. Daily during presumed gestation and on Day 1 postpartum (rats assigned to natural delivery). Sacrifice: Terminal weight. Feed Consumption Values (recorded and tabulated): Dosage Period: Weekly to cohabitation. On days 0, 7, 14, 21 and 25 (if necessary) of presumed gestation and on days 1 and 5 postpartum (rats assigned to natural delivery). Feed consumption values may be recorded more frequently than cited above if it is necessary to replenish the feed. During cohabitation, when two rats occupy the same cage with one feed jar, replenishment of the feed jars will be documented. Individual values will not be recorded or tabulated. 000403 418-018:PAGE C-13 Mating Performance: Protocol 418-018 Page 13 Mating will be evaluated daily during the cohabitation period and confirmed by observation of spermatozoa in a smear of the vaginal contents and/or a copulatory plug observed in situ. Fertility Parameters: Fertility Index (percentage of matings that result in pregnancies). Gestation Index (percentage of pregnancies that result in birth of live litters). Number of offspring per litter (live and dead pups). Number of implantation sites. General condition of dam and litter during the postpartum period. Viability Indices (percentage of pups bom that survive 5 days). Caesarean-Sectioning Observations: Eight rats from dosage groups 1 to 7, 12 and 13 (Replicate 1) will be Caesareansectioned on day 21 of presumed gestation. The fetuses will be removed from the uterus and pooled by litter for body weights. The rats will be examined for number and distribution of: Corpora Ltea. Implantation Sites. [Placentae that appear abnormal (size, color or shape) will be noted in the raw data]. Live and Dead Fetuses. (A live fetus is defined as one that responds to stimuli; a dead fetus is defined as a term fetus that does not respond to stimuli and that is not markedly autolyzed; dead fetuses demonstrating marked to extreme autolysis are considered to be late resorptions.) Early and Late Resorptions. (A conceptus is defined as a late resorption if it is grossly evident that organogenesis has occurred; if this is not the case, the conceptus is defined as an early resorption.) 000494 418-018:PAGE C-14 Fetal Observations: Protocol 418-018 Page 14 Body Weights: Fetuses will be pooled by litter and litter body weights will be recorded. (Only body weights of live fetuses will be recorded.) Blood and Tissue Samples (See ATTACHMENT 4 for tissue collection and processing): Caps and labeled tubes will be weighed (combined weight, to the nearest .001 gram) before (empty) and after retention of pooled fetus samples for subsequent use in pharmacokinetic analyses. These weights will be documented in the raw data, and copies of these weights will be included with the packing list prior to shipment. Sample tubes will be labeled with the study number, litter identification, date of collection, study day, sample identification and collection timepoint. Blood samples will be collected from each fetus via decapitation. Blood from approximately one-half of the fetuses in each litter will be placed into EDTA-coated tubes and the remaining fetal blood will be transferred into serum separator tubes. The samples will be spun in a centrifuge and the resulting serum (divided into two aliquots)/plasma (one aliquot) will be transferred into labeled polypropylene tubes. All samples will be immediately frozen on dry ice and maintained frozen (<-20C) until shipment for analysis. The liver from each fetus will be collected. One fetal liver from each litter will be removed and fixed in gluteraldehyde for possible future analysis by electron microscopy. The remaining fetal livers in each litter will be divided into three samples of equal number (when possible). Two of the samples will be frozen and stored (<-20C) until shipment for analysis. The remaining sample will be flash frozen in liquid nitrogen and maintained frozen (<-70C) until shipment for possible analysis. Carcasses will be discarded without further evaluation. Sample Analysis and Shipping Instructions: Appropriate samples will be maintained frozen until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. One of each pair of serum samples, frozen liver samples, and liver samples fixed in gluteraldehyde will be sent to Dave Ehresman, at the previously cited address. The sample tube and liver weights and a packing list will be included with the samples sent to Dave Ehresman. 000405 418-018:PAGE C-l Protocol 418-018 Page 15 The remaining serum and liver samples will be analyzed for LDL, HDL, total cholesterol and triglycerides. These samples will be shipped to: Dr. Saroj R. Das Ani Lytics, Inc. 200 Gerard Street, Suite 200 Gaithersburg, Maryland 20877 Telephone: (301)921-0168 Telefax: (301)977-0433 Plasma mevalonic acid levels will be measured. These samples will be shipped to: Jim Jersey Primedica Worcester Laboratories 57 Union Street Worcester, Massachusetts 01608 Telephone: (508)890-0151 Telefax: (508)753-1834 Both the recipients and the Study Monitor will be notified in advance of sample shipment. Natural Delivery: Female rats will be evaluated for: Adverse Clinical Signs Observed During Parturition. Duration of Gestation (day 0 of presumed gestation to the time the first pup is observed). Litter Size (defined as all pups delivered). Pup Viability at Birth. METHOD OF SACRIFICE - Fo GENERATION: Rats will be sacrificed by carbon dioxide asphyxiation. Fetuses will be sacrificed via decapitation. When possible, rats, pups and fetuses will be sacrificed and tissues collected at approximately the same time each day. 000406 418-018:PAGE C-16 NECROPSY - Fo GENERATION: Protocol 418-018 Page 16 Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation (a table of random units will be used to select one control group rat assigned to Caesarean-section from which all tissues examined at necropsy will be retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions). Unless specifically cited below, all other tissues will be discarded. Blood and Tissue Sample Collection (See ATTACHMENT 4 for tissue collection and processing): Caps and labeled tubes will be weighed (combined weight, to the nearest .001 gram) before (empty) and after retention of samples for subsequent use in pharmacokinetic analyses. These weights will be documented in the raw data, and copies of these weights will be included with the packing list prior to shipment. Sample tubes will be labeled with the study number, animal identification, date of collection, study day, sample identification and collection timepoint. Female Rats Assigned to Caesarean-Sectioning: On day 21 of presumed gestation, blood and liver samples will be collected from the eight rats designated for Caesarean-section in dosage groups 1 to 7, 12 and 13. The time of sample collections will be recorded in the raw data. Blood samples (at least 4 mL each) will be collected from the rats via the inferior vena cava and separated into two approximately equal aliquots. The first aliquot will be placed into EDTA-coated tubes and the second aliquot will be transferred into serum separator tubes. The samples will be spun in a centrifuge and the resulting serum (divided into two aliquots)/plasma (one aliquot) will be transferred into labeled polypropylene tubes. All samples will be immediately frozen on dry ice and maintained frozen (<-20C) until shipment for analysis. Following collection of blood samples, the liver of each rat will be excised and the liver weight will be recorded. A portion of each liver sample (right lateral lobe) will be flash frozen in liquid nitrogen and maintained frozen (<-70C) until shipment for possible analysis. The median liver lobe will be frozen and stored (<-20C) until shipment for analysis. A section (approximately 0.3 to 0.5 cm wide) from the remaining portion of the liver will be fixed in gluteraldehyde for possible future analysis by electron microscopy. The remaining portion of the liver will be frozen and stored (<-20C) until shipment for analysis. Female Rats Assigned to Natural Delivery: Blood, milk, heart and liver samples will be collected from maternal rats on day 5 postpartum. Time of sample collections (blood and milk) will be recorded in the raw data. 000407 418-018:PAGE C-17 Protocol 418-018 Page 17 On day 5 postpartum, each dam will be removed from the nesting box and individually housed for approximately four hours. The dam will be injected intravenously with one unit of oxytocin approximately five minutes before milk samples (at least 100 pL per sample) are collected. The samples will be immediately frozen on dry ice and maintained frozen (<-20C) until shipment for analysis. Following milk sample collection, blood samples (at least 4 mL each) will be collected from the rats via the inferior vena cava and separated into two approximately equal aliquots. The first aliquot will be placed into EDTA-coated tubes and the second aliquot will be transferred into serum separator tubes. The samples will be spun in a centrifuge and the resulting serum (divided into two aliquots)/plasma (one aliquot) will be transferred into labeled polypropylene tubes. All samples will be immediately frozen on dry ice and maintained frozen (<-20C) until shipment for analysis. Following collection of milk and blood samples, the heart and liver of each dam will be collected. The liver weight will be recorded. A portion of each liver sample (right lateral lobe) will be flash frozen in liquid nitrogen and maintained frozen (<-70C) until shipment for possible analysis. The median liver lobe will be frozen and stored (<-20C) until shipment for analysis. The heart will be excised and two cuts will be made to allow proper fixation. The first cut will start to the right of the ventral midline surface at the apex and extend anteriorly and ventrally to the pulmonary artery. The second cut will be made starting to the left of the ventral midline surface at the apex and extend through the left ventricle to the ascending aorta. The cut heart and a section (approximately 0.3 to 0.5 cm wide) from the remaining portion of the liver will be fixed in gluteraldehyde for possible future analysis by electron microscopy and/or light microscopy. The remaining portion of the liver will be frozen and stored (<-20C) until shipment for analysis. Sample Analysis and Shipping Instructions: Appropriate samples will be maintained frozen until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. One of each pair of serum samples, frozen maternal liver samples (right lateral and median lobes), and liver and heart samples fixed in gluteraldehyde will be sent to Dave Ehresman, at the previously cited address. The sample tube and liver weights and a packing list will be included with the samples sent to Dave Ehresman. 000408 418-018:PAGE C-18 Protocol 418-018 Page 18 The remaining serum and liver samples will be analyzed for LDL, HDL, total cholesterol and triglycerides. Milk samples will be analyzed for total cholesterol. These samples will be shipped to: Dr. Saroj R. Das Ani Lytics, Inc. 200 Gerard Street, Suite 200 Gaithersburg, Maryland 20877 Telephone: (301)921-0168 Telefax: (301)977-0433 Plasma mevalonic acid levels will be measured. These samples will be shipped to: Jim Jersey Primedica Worcester Laboratories 57 Union Street Worcester, Massachusetts 01608 Telephone: (508)890-0151 Telefax: (508)753-1834 Both the recipients and the Study Monitor will be notified in advance of sample shipment. Scheduled Sacrifice - Female Rats Assigned to Caesarean-Sectioning: On day 21 of presumed gestation, female rats will be sacrificed, Caesarean-sectioned, and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. Blood and tissue samples (blood serum/plasma and liver) will be collected as previously described. Uteri of apparently nonpregnant rats will be stained with 10% ammonium sulfide to confirm the absence of implantation sites(5) and retained in neutral buffered 10% formalin for possible future evaluation. Scheduled Sacrifice - Female Rats Assigned to Natural Delivery: On day 5 of lactation, female rats will be sacrificed and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. Blood and tissue samples (blood serum/plasma, heart and liver) will be collected as previously described. Rats that do not deliver a litter will be sacrificed on day 25 of presumed gestation and examined for gross lesions. Blood and tissue samples will not be collected. Uteri will be stained with 10% ammonium sulfide to confirm the absence of implantation sites(5) and retained in neutral buffered 10% formalin for possible future evaluation. 000409 4 18-01 S:PAGE C -19 Dams with No Surviving Pups: Protocol 418-018 Page 19 Dams with no surviving pups will be sacrificed after the last pup is found dead, missing or presumed cannibalized. Blood and tissue samples (blood serum/plasma, heart and liver) will be collected as previously described. A gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. Postpartum data for these dams will be excluded from summary tables. Rats Found Dead or Moribund: Rats that die or are sacrificed because of moribund condition or abortion will be examined for the cause of death or moribund condition on the day the observation is made. The rats will be examined for gross lesions. When possible (not precluded by autolysls), blood serum/plasma, heart and liver samples will be retained as previously described for rats assigned to natural delivery. Pregnancy status and uterine contents of female rats will be recorded. Aborted fetuses and/or delivered pups will be examined to the extent possible. Uteri of apparently nonpregnant rats will be stained with 10% ammonium sulfide to confirm the absence of implantation sites(5) and retained in neutral buffered 10% formalin for possible future evaluation. TESTS, ANALYSES AND MEASUREMENTS - F1 GENERATION: Viability: Postpartum Period: Litters will be observed for dead pups at least twice daily. The pups in each litter will be counted once daily. Clinical Observations and/or General Appearance: Postpartum Period: Clinical observations and sex will be recorded once daily. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or the Study Monitor. Body Weights: Postpartum Period: Days 1 (birth), 2, 3, 4 and 5 postpartum. Pooled litter weights will be recorded. METHOD OF SACRIFICE - F1 GENERATION PUPS: As previously cited for Fo generation rats. When possible, pups will be sacrificed and tissues collected at approximately the same time each day. The time of sample collection will be recorded. 000410 418-018:PAGE C-20 F1 Generation Pups Found Dead on Day 1 Postpartum: Protocol 418-018 Page 20 Pups that die before examination of the litter for pup viability will be evaluated for vital status at birth. The lungs will be removed and immersed in water. Pups with lungs that sink will be identified as stillborn; pups with lungs that float will be identified as livebom, and to have died shortly after birth. Pups with gross lesions will be preserved in Bouin's solution for possible future evaluation. Should postmortem autolysis preclude these evaluations, it will be noted in the necropsy data. F1 Generation Pups Found Dead or Moribund on Days 2 to 5 Postpartum: Pups found dead or sacrificed due to moribund condition will be examined for gross lesions and for the cause of the moribund condition or death. Pups with gross lesions found on days 2 to 4 postpartum will be preserved in Bouin's solution for possible future evaluation; gross lesions of pups found on day 5 postpartum will be preserved in neutral buffered 10% formalin. Should postmortem autolysis preclude these evaluations it will be noted in the necropsy data. Scheduled Sacrifice - F1 Generation Pups (See ATTACHMENT 4 for tissue collection and processing): On day 5 postpartum, pups will be sacrificed and examined for gross lesions. Gross lesions will be preserved in neutral buffered 10% formalin. Necropsy will include a single cross-section of the head at the level of the frontal-parietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Caps and labeled tubes will be weighed (combined weight, to the nearest .001 gram) before (empty) and after retention of samples for subsequent use in pharmacokinetic analyses. These weights will be documented in the raw data, and copies of these weights will be included with the packing list prior to shipment. Sample tubes will be labeled with the study number, animal identification, date of collection, study day, sample identification and collection timepoint. Blood samples will be collected via cardiac puncture from each pup, pooled (by sex per litter) and separated into four (two per sex) approximately equal aliquots. The first aliquot per sex will be placed into EDTA-coated tubes and the second aliquot per sex will be transferred into serum separator tubes. The samples will be spun in a centrifuge and the resulting serum (divided into two aliquots)/plasma (one aliquot) will be transferred into labeled polypropylene tubes. All samples will be immediately frozen on dry ice and maintained frozen (<-20C) until shipment for analysis. 000411 418-018:PAGE C-21 Protocol 418-018 Page 21 The liver from each pup will be collected, pooled (by sex per litter) and the pooled weight recorded. One liver from each litter pool will be removed and fixed in gluteraldehyde for possible future analysis by electron microscopy. The remaining livers in each litter pool will be divided into three samples of equal number (when possible). Two of the samples will be frozen and stored (<-20C) until shipment for analysis. The remaining sample will be flash frozen in liquid nitrogen and maintained frozen (-70C) until shipment for possible analysis. The hearts will be collected from the first two male and two female pups from each litter (pooled by sex per litter) and will be fixed in gluteraldehyde for possible future analysis by electron microscopy and/or light microscopy. The remaining carcasses will be discarded without further evaluation Samples will be shipped for analysis as previously described for dams and fetuses. 000412 418-018:PAGE C-22 PROPOSED STATISTICAL METHODS*612): Protocol 418-018 Page 22 Averages and percentages will be calculated. Litter values will be used where appropriate. Additional procedures and/or analyses may be performed, if appropriate. Type of Test3 I. Parametric Nonparametricb A. Bartlett's Test0 A. Kruskal-Wallis Test (<75^o ties) Significant at p<0.001 Not Significant Significant at p<0.05 Not Significant Nonparametric Analysis of Variance Dunn's Test Significant at p<0.05 Not Significant B. Fisher's Exact Test (>75% ties) Dunnett's Test III. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilities are reported as either p<0.05 or p i 0.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. 000413 418-018:PAGE C-23 DATA ACQUISITION, VERIFICATION AND STORAGE: Protocol 418-018 Page 23 Data generated during the course of this study will be recorded either by hand or using the Primedica Argus Automated Data Collection and Management System and the Vivarium Temperature and Relative Humidity Monitoring System. All data will be tabulated, summarized and/or statistically analyzed using the Primedica Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microsoft Excel [part of Microsoft Office 97 (version SR-2)] and/or The SAS System (version 6.12). Records will be reviewed by the Study Director and/or appropriate management personnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft final report, after which time the Sponsor will be contacted to determine the disposition of these materials. RECORDS TO BE MAINTAINED: Protocol and Amendments. Test and Control Article, Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (if prescribed by Staff Veterinarian). General Comments. Clinical Observations and/or General Appearance. Pharmacokinetic Sample Collection, Processing and Shipment. Body Weights. Feed Consumption Values. Caesarean-Sectioning Observations. Natural Delivery Observations. Litter Observations. Gross Necropsy Observations. Organ Weights. Photographs (if required). Study Maintenance (room and environmental records). Feed, Water and Bedding Analyses. Packing and/or Shipment Lists. 000414 418-018-.PAGEC-24 KEY PERSONNEL: Protocol 418-018 Page 24 Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Director of Research: Alan M. Hoberman, Ph.D., DABT Associate Director of Research and Study Director: Raymond G. York, Ph.D., DABT Director of Operations and Compliance: Barbara J. Patterson, B.A. Director of Laboratory Operations: John F. Barnett, B.S. Director of Study Management: Valerie A. Sharper, M.S. Manager of Animal Operations and Chairperson, Institutional Animal Care and Use Committee: Dena C. Lebo, V.M.D. Consultant, Veterinary Pathology: W. Ray Brown, D.V.M., Ph.D., ACVP FINAL REPORT: A comprehensive draft final report will be prepared on completion of the study and will be finalized following consultation with the Sponsor. The report will include the following: Summary and Conclusion. Experimental Design and Method. Evaluation of Test Results. Appendices: Figures, Summary and Individual Tables Summarizing the Above Data, Protocol and Associated Amendments and Deviations, Study Director's GLP Compliance Statement, Reports of Supporting Data (if appropriate) and QAU Statement. The Sponsor will receive one copy of the draft report and two copies of the final report. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described in this protocol have been reviewed by the Testing Facility's Institutional Animal Care and Use Committee. All procedures described in this protocol that involve study animals will be conducted in a manner to avoid or minimize discomfort, distress or pain to the animals. The Sponsor's signature below documents the fact that information concerning the necessity for conducting this study and the fact that this is not an unnecessarily duplicative study may be obtained from the Sponsor. No alternative (in vitro) procedures were available for meeting the stated purposes of the study. 000415 418-018 :PAGE C-25 REFERENCES: Protocol 418-018 Page 25 1. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 2. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November?, 1983), J. Clin. Psychiat. 45(9):7-10. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River CrhCD^BR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 4. Institute of Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski, E. (1964). Farbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. 6. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 240-241. 7. Sokai, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. 8. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 9. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-1121. 10. Sokai, R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 11. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241-252. 12. Siegel, S. (1956). Nonparametric Statistics for the Behavioral Sciences, McGraw-Hill, New York, pp. 96-104. 000416 PROTOCOL APPROVAL: FOR THE TESTING FACILITY Dearlove, Ph.D., DABT Associate Director of Research 418-018:PAGE C-26 Protocol 418-018 Page 26 Date ^ ^ r - . . C . . . 4 r ^ 7 r r . _____________ Dena C. Lebo, V.M.D. Chairperson, Institutional Animal Care and Use Committee FOR THE SPONSOR j Q Deanna J. LuebK^r, M.S. Study Monitor John Butenhoff, Ph.D., DABT, CIH Alternate Study Monitor ____ Date .>____ Date 2 3- A Date - Zf& O 000417 418-018:PAGE C-27 ATTACHMENT 1 STUDY SCHEDULE 000418 ATTACHMENT 1 418-018:PAGE C-28 Protocol 418-018 Page 1 of 2 SCHEDULE3 REPLICATE 1b 22 AUG 00 Animal Receipt - Acclimation Begins (Female Rats). 29 AUG 0 0 -1 2 NOV 00 29 AUG 00 - 20 NOV 00 Dosage Period - Female Rats Assigned to Caesarean-Sectioning (42 days before cohabitation and continuing through day 20 of presumed gestation). Dosage Period - Female Rats Assigned to Natural Delivery [42 days before cohabitation through day 24 of presumed gestation (rats that do not deliver a litter) or day 4 postpartum (rats that deliver a litter)]. 09 OCT 00 PM - 16 OCT 00 AM 16 OCT 00 PM - 23 OCT 00 AM Cohabitation Period (Maximum of 14 days). Male 1 (07 days) Male 2 (07 days) 10 OCT 00 23 OCT 00 First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. 31 OCT 00 13 NOV 00 First Possible Day 21 of Presumed Gestation Caesarean-sectioning. Last Possible Day 21 of Presumed Gestation Caesarean-sectioning. 31 OCT 00 17 NOV 00 First Possible Delivery (Day 21 of presumed gestation). Last Possible Delivery (Day 25 of presumed gestation). 04 NOV 00 17 NOV 00 First Possible Day 25 of Presumed Gestation Female Sacrifice. Last Possible Day 25 of Presumed Gestation Female Sacrifice. a. The study initiation date is the day the Study Director signs the protocol. b. Replicate 2 will be initiated two weeks after the start of Replicate 1 (see Page 2 of ATTACHMENT 1). 000419 ATTACHMENT 1 418-018:PAGE C-29 Protocol 418-018 Page 2 of 2 04 NOV 00 21 NOV 00 First Possible Day 5 Sacrifice (Dams and F1 generation pups). Last Possible Day 5 Sacrifice. REPLICATE 2 05 SEP 00 Animal Receipt - Acclimation Begins (Female Rats). 14 SEP 0 0 - 0 6 DEC 00 Dosage Period - Female Rats Assigned to Natural Delivery [42 days before cohabitation through day 24 of presumed gestation (rats that do not deliver a litter) or day 4 postpartum (rats that deliver a litter)]. 25 OCT 00 PM - 01 NOV 00 AM 01 NOV 00 PM - 08 NOV 00 AM Cohabitation Period (Maximum of 14 days). Male 1 (07 days) Male 2 (07 days) 26 OCT 00 08 NOV 00 First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. 16 NOV 00 03 DEC 00 First Possible Delivery (Day 21 of presumed gestation). Last Possible Delivery (Day 25 of presumed gestation). 20 NOV 00 03 DEC 00 First Possible Day 25 of Presumed Gestation Female Sacrifice. Last Possible Day 25 of Presumed Gestation Female Sacrifice. 20 NOV 00 07 DEC 00 First Possible Day 5 Sacrifice (Dams and F1 generation pups). Last Possible Day 5 Sacrifice. 01 MAR 01 Draft Final Report. 000420 418-018:PAGH C-30 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 000421 i ir r\w w m i DATA SHEET 3M Center St. Paul, Minnesota 55144-1000 1-800-364-3577 or (612) 737-6501 (24 hours) 418-018:PAGE C-31 Copyright, 1998, Minnesota Mining and Manufacturing Company. All rights reserved. Copying and/or downloading of this information for the purpose of properly utilizing 3M products is allowed provided that: 1) the inforaation is copied in full with no changes unless prior agreement is obtained from 3M, and 2) neither the copy nor the original is resold or otherwise distributed with the intention of earning a profit thereon. DIVISION: 3M CHEMICALS TRADE NAME: FC-95 FLU0RAD Brand Fluorocheaical Surfactant ID NUMBER/U.P.C .: 98-0207-0103-7 00-51135-09054-1 98-0207-0104-5 98-0211-0888-5 00-51135-09362-7 98-0211-3916-1 Z F -0002-1044 -1 ISSUED: January 29, 1998 SUPERSEDES: November 05, 1997 DOCUMENT: 10-3796-9 00-51135-09055-8 00-51135-02311-2 1. INGREDIENT C.A.S. NO. PERCENT POTASSIUM PERFLUOROALKYL SULFONATE___ POTASSIUM PERFLUOROALKYL SULFONATE___ POTASSIUM PERFLUOROALKYL SULFONATE___ POTASSIUM PERFLUOROALKYL SULFONATE___ POTASSIUM PERFLUOROALKYL SULFONATE___ 2795-39-3 3871-99-6 29420-49-3 60270-55-5 3872-25-1 82 3 3 2 1 - 86 -8 -7 -6 -3 2. PHYSICAL DATA BOILING POINT:...... VAPOR PRESSURE:.... VAPOR DENSITY:..... EVAPORATION RATE:... SOLUBILITY IN WATER: SPECIFIC GRAVITY:... PERCENT VOLATILE:.. . P H : ................ VISCOSITY:......... MELTING POINT:..... N/A N/A N/A N/A slight ca. 0.6 Water=1 (Bulk) 0% 7 -8 (0.1% Aqueous) N/D N/O APPEARANCE AND ODOR: Light colored, free flowing powder. 000422 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately USDS: FC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 418-018:PAGE C-32 PAGE 2 3. FIRE AND EXPLOSION HAZARD DATA FLASH POINT:.................. None FLAMMABLE LIMITS -LEL:........ N/A FLAMMABLE LIMITS -UEL:........ N/A AUTOIGNITION TEMPERATURE:..... N/A EXTINGUISHING MEDIA: Mater, Carbon dioxide, Dry chemical, Foam SPECIAL FIRE FIGHTING PROCEDURES: Wear full protective clothing, including helmet, self-contained, positive pressure or pressure demand breathing apparatus, bunker coat and pants, bands around arms, waist and legs, face mask, and protective covering for exposed areas of the head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See Hazardous Decomposition section for products of combustion. 4. REACTIVITY DATA STABILITY: Stable INCOMPATIBILITY - MATERIALS/CONDITIONS TO AVOID: Not applicable. HAZARDOUS POLYMERIZATION: Hazardous polymerization will not occur. HAZARDOUS DECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Sulfur, Hydrogen Fluoride, Toxic Vapors, Gases or Particulates. 5. ENVIRONMENTAL INFORMATION SPILL RESPONSE: Observe precautions from other sections. Vacuum, use wet sweeping compound or water to avoid dusting. CAUTION! A vacuum cleaner could be an ignition source. Clean up residue with water. Place in an approved metal container. Seal the container. RECOMMENDED DISPOSAL: Do not release to waterways or sewer. Do not use in products or processes that could result in aquatic concentrations greater than 1/10 of the lowest EC50 or LC50 concentration. Incinerate in an industrial or commercial facility in the presence of a combustible material. Combustion products will include HF. Disposal alternative: Dispose of waste product in a facility permitted to 000423 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately USDS: FC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 418-018:PAGE C-33 PAGE 3 5. ENVIRONMENTAL INFORMATION (continued) accept chenical waste. ENVIRONMENTAL DATA: 96-Hr. Aquatic Fish LC50, Fathead Minnow(Pimephales promelas)=38 mg/1, Bluegill Sunfish(Lepomis macrochirus)*68 ag/1, Rainbow Trout(Salmo gairdneri)=11 mg/1; 48-Hr. EC50, Daphnia Magna = 50 ag/1; COD=.004 g/g; BOD20 = Nil. REGULATORY INFORMATION: Volatile Organic Compounds: N/A. VOC Less H20 & Exempt Solvents: N/A. Since regulations vary, consult applicable regulations or authorities before disposal. U.S. EPA Hazardous Waste Number = None (Not U.S. EPA Hazardous). This product complies with the chemical registration requirements of TSCA, EINECS, CDSL, AICS, MIT1 and Korea. EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes 6. SUGGESTED FIRST AID EYE CONTACT: Immediately flush eyes with large amounts of water for at least 15 minutes. Get immediate medical attention. SKIN CONTACT: Immediately flush skin with large amounts of water. Remove contaminated clothing. If irritation persists, call a physician. Wash contaminated clothing before reuse. INHALATION: If signs/symptoms occur, remove person to fresh air. If signs/symptoms continue, call a physician. IF SWALLOWED: Drink two glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION EYE PROTECTION: Avoid eye contact. Wear vented goggles. 000424 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately USDS: FC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 418-018:PAGE C-34 PAGE 4 7. PRECAUTIONARY INFORMATION (continued) SKIN PROTECTION: Avoid skin contact. Wear appropriate gloves when handling this aterial. A pair of gloves eade froe the following Material(s) are recommended: butyl rubber. Use one or more of the following personal protection items as necessary to prevent skin contact: head covering, coveralls. Protective garments (other than gloves) should be made of either of the following materials: polyethylene/polyvinylidene chloride (Saranex). RECOMMENDED VENTILATION: Use with appropriate local exhaust ventilation. Use in a wellventilated area. Provide sufficient ventilation to maintain emissions below recommended exposure limits. If exhaust ventilation is not adequate, use appropriate respiratory protection. RESPIRATORY PROTECTION: Avoid breathing of dust. Select one of the following NIOSH approved respirators based on airborne concentration of contaminants and in accordance with OSHA regulations: half-mask dust and mist respirator, half-mask supplied air respirator, full-face dust and mist respirator, full-face supplied air respirator. PREVENTION OF ACCIDENTAL INGESTION: Do not eat, drink or smoke when using this product. Wash exposed areas thoroughly with soap and water. Wash hands after handling and before eating. RECOMMENDED STORAGE: Keep container dry. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: Nonflammable. OTHER PRECAUTIONARY INFORMATION: No smoking: Smoking while using this product can result in contamination of the tobacco and/or smoke and lead to the formation of the hazardous decomposition products mentioned in section 4 of this MSDS. HMIS HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.) INGREDIENT EXPOSURE LIMITS VALUE UNIT TYPE AUTH SKIN POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 TWA 3M TWA 3M TWA 3M TWA 3M Y Y Y Y Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 000425 USDS: FC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 EXPOSURE LIMITS (continued) 418-018:PAGE C-35 PAGE 5 INGREDIENT VALUE UNIT TYPE AUTH SKIN* POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 TWA 3M Y * SKIN NOTATION: Listed substances indicated with 'Y' under SKIN refer to the potential contribution to the overall exposure by the cutaneous route including aucous aenbrane and eye, either by airborne or, aore particularly, by direct contact with the substance. Vehicles can alter skin absorption. SOURCE OF EXPOSURE LIMIT DATA: - 3M: 3M Recoamended Exposure Guidelines 8. HEALTH HAZARD DATA EYE CONTACT: Mild Eye Irritation: signs/syaptoas can include redness, swelling, pain, and tearing. SKIN CONTACT: Mild Skin Irritation (after prolonged or repeated contact): signs/syaptoas can include redness, swelling, and itching. May be absorbed through the skin and persist in the body for an extended tiae. INHALATION: May be harmful if inhaled. May be absorbed by inhalation and persist in the body for an extended time. Single overexposure, above recoamended guidelines, may cause: Irritation (upper respiratory): signs/symptoms can include soreness of the nose and throat, coughing and sneezing. IF SWALLOWED: Ingestion is not a likely route of exposure to this product. Illness may result from a single swallowing of a moderate quantity of this material. May be harmful if swallowed. MUTAGENICITY: Mutagenicity assays indicate the product is not mutagenic. 000426 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately USDS: FC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 418-018:PAGE C-36 PAGE 6 8. HEALTH HAZARD DATA (continued) REPRODUCTIVE/DEVELOPMENTAL TOXINS: Not teratogenic in the rat at oral doses below Maternally toxic levels. OTHER HEALTH HAZARD INFORMATION: This product is not known to contain any substances regulated under California Proposition 65. A Product Toxicity Summary Sheet is available. SECTION CHANGE DATES HEADING SECTION CHANGED SINCE November 05, 1997 ISSUE Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately The information in this Material Safety Data Sheet (MSDS) is believed to be correct as of the date issued. 3M MAKES NO WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User is responsible for determining whether the 3M product is fit for a particular purpose and suitable for user's method of use or application. Given the variety of factors that can affect the use and application of a 3M product, some of which are uniquely within the user's knowledge and control, it is essential that the user evaluate the 3M product to determine whether it is fit for a particular purpose and suitable for user's method of use or application. 3M provides information in electronic form as a service to its customers. Due to the remote possibility that electronic transfer may have resulted in errors, omissions or alterations in this information, 3M makes no representations as to its completeness or accuracy. In addition, information obtained from a database may not be as current as the information in the MSDS available directly from 3M. 000427 418-018.-PAGE C-37 PRODUCT SA AMK A^C. C T '6' ^ - 7 ^ 5 0 Snruce Street #: C8503 NAME; CHOLESTEROISum Louis. Missouri S3HJ3 USA ; Y J | W r A SHEET, Valid 8/2000 - &>Bpiaf-52l-MS6 (3i> 771-5765 rcJ ^ d 07/25/2000 15:03 Fmx800-325-5052 *(314) 771-5757 email: tio-a)d8siat.cBm nru.llnia-aidncfi.com 3050 Spruce Street St. Louis, MO 63103, US Technical Phone; 314 771 5765 Emergency Phone: 314 111 5765 Fax: 800 325 5052 i fOM-t ( I k tA- SECTION 1. - - - - - - - - - CHEMICAL IDENTIFICATION- - - - CATALOG #: C8503 NAME: CHOLESTEROL SECTION 2. - - - - - COMPOSITION/INFORMATION ON INGREDIENTS CAS #: 57-88-5 MF: C27H460 EC NO: 200-353-2 SYNONYMS CHOLEST-5-EN-3-OL (3-BETA)- (9CI) * CHOLEST-5-EN-3-BETA-OL ' DELTA[SUP 5)-CHOLESTEN-3-BETA-OL * 5-CHOLESTEN-3-3ETA-OL * 5:6- CHOLESTEN-3-OL * 5:6-CHOLESTEN-3-BETA-OL * CHOLESTERIN * CHOLESTERINE * CHOLESTRIN * (-)-CHOLESTEROL * CHOLESTERYL ALCOHOL * DYTHOL * 3- BETA-HYDROXYCHOLEST-5-ENE * PROVITAMIN D * SECTION 3. - - - - - - - - - - HAZARDS IDENTIFICATION - - - - - - - - - DATA NOT AVAILABLE SECTION 4. - - - - - - - - - - FIRST-AID MEASURES- - - - - - - - - - - IN CASE OF CONTACT, IMMEDIATELY FLUSH EYES WITH COPIOUS AMOUNTS OF WATER FOR AT LEAST 15 MINUTES. IN CASE OF CONTACT, IMMEDIATELY WASH SKIN WITH SOAP AND COPIOUS AMOUNTS OF WATER. IF INHALED, REMOVE TO FRESH AIR. IF NOT BREATHING GIVE ARTIFICIAL RESEIRATION. IF BREATHING IS DIFFICULT, GIVE OXYGEN. IF SWALLOWED, WASH OUT MOUTH WITH WATER PROVIDED PERSON IS CONSCIOUS. CALL A PHYSICIAN. WASH CONTAMINATED CLOTHING BEFORE REUSE. SECTION 5. - - - - - - - - - FIRE FIGHTING MEASURES - - - - - - - - - - EXTINGUISHING MEDIA WATER SPRAY. CARBON DIOXIDE, DRY CHEMICAL POWDER OR APPROPRIATE FOAM. SPECIAL FIREFIGHTING PROCEDURES WEAR SELF-CONTAINED BREATHING APPARATUS AND PROTECTIVE CLOTHING TO PREVENT CONTACT WITH SKIN AND EYES. UNUSUAL FIRE AND EXPLOSIONS HAZARDS EMITS TOXIC FUMES UNDER FIRE CONDITIONS. SECTION 6. - - - - - - - - ACCIDENTAL RELEASE MEASURES- - - - - - - - - CHEMICAL SAFETY GOGGLES. USE PROTECTIVE CLOTHING, GLOVES AND MASK. SWEEP UP. PLACE IN A BAG AND HOLD FOR WASTE DISPOSAL. AVOID RAISING DUST. VENTILATE AREA AND WASH SPILL SITE AFTER MATERIAL PICKUP IS COMPLETE. SECTION 7. - - - - - - - - -- HANDLING AND STORAGE- - -- - REFER TO SECTION 8. SECTION 8. - - - - - - EXPOSURE CONTROLS/PERSONAL PROTECTION CHEMICAL SAFETY GOGGLES. COMPATIBLE CHEMICAL-RESISTANT GLOVES. NIOSH/MSHA-APPROVED RESPIRATOR. 000428 Page 1 We art cam m i: ted ce the S u c c c j j ej out C uctamen. Employee and S h a r e h o l d e r , c h r o u fh L i f e S e n n e t . T e c h n o l oj j and Ser vi e, _____ ______ .... __ __ ___ _ ____ _ 418-018:PAGE C-38 3050 Spivce srre; PRODUCT #: C05O3 NAME: CHOLESTEROLsammous. Missouri 63103 USA lS A S ^ Y J^ifA SHEET, Valid 8/2000 - left^iftafib-SEi-BSSe-(314) 771.5765 td 07/25/2000 15:03 Pax800J25-50S2(3i4)771-5757 ______________________ ____________________ amai!- mp-alrtaa! c o m _________ wwwsigma-attnen.eon' SHOWER AN? EYE BATH _ MECHANICAL EXHAUST REQUIRED. AVOID INHALATION. AVOID CONTACT WITH EYES, SKIN AND CLOTHING. AVOID PROLONGED OR REPEATED EXPOSURE. WASH THOROUGHLY AFTER HANDLING. KEEP TIGHTLY CLOSED. STORE IN A COOL DRY PLACE. SECTION 9. - - - - - - - PHYSICAL AND CHEMICAL PROPERTIES - - - - - - - APPEARANCE AND ODOR WHITE POWDER PHYSICAL PROPERTIES BOILING POINT: 360 C MELTING POINT: 147 C TO 149 C SPECIFIC GRAVITY: 1.067 SECTION 10. - - - - - - - - -STABILITY AND REACTIVITY - - - - - - - - - STABILITY STABLE. INCOMPATIBILITIES STRONG OXIDIZING AGENTS HAZARDOUS COMBUSTION OR DECOMPOSITION PRODUCTS TOXIC FUMES OF: CARBON MONOXIDE. CARBON DIOXIDE HAZARDOUS POLYMERIZATION WILL NOT OCCUR. SECTION 11. - - - - - - - - - TOXICOLOGICAL INFORMATION - - - - - - - - ACUTE EFFECTS MAY BE HARMFUL BY INHALATION, INGESTION, OR SKIN ABSORPTION. MAY CAUSE IRRITATION. TOXICOLOGICAL DATA INDICATE A LOW ORDER OF TOXICITY. GENERAL GOOD HOUSEKEEPING PRACTICES SHOULD BE FOLLOWED. RTECS #: FZ8400000 CHOLESTEROL TARGET ORGAN DATA KIDNEY, URETER, BLADDER (BLADDER TUMORS) EFFECTS ON FERTILITY (POST-IMPLANTATION MORTALITY) EFFECTS ON FERTILITY (LITTER SIZE) SPECIFIC DEVELOPMENTAL ABNORMALITIES (CRANIOFACIAL) TUMORIGENIC (EQUIVOCAL TUMORIGENIC AGENT BY RTECS CRITERIA) TUMORIGENIC (TUMORS AT SITE OF APPLICATION) ONLY SELECTED REGISTRY OF TOXIC EFFECTS OF CHEMICAL SUBSTANCES (RTECS) DATA IS PRESENTED HERE. SEE ACTUAL ENTRY IN RTECS FOR COMPLETE INFORMATION. SECTION 12. - - - - - - - - - ECOLOGICAL INFORMATION - - - - - - - - - DATA NOT YET AVAILABLE. SECTION 13. - - - - - - - - - DISPOSAL CONSIDERATIONS - - - - - - - - - DISSOLVE OR MIX THE MATERIAL WITH A COMBUSTIBLE SOLVENT AND BURN IN A CHEMICAL INCINERATOR EQUIPPED WITH AN AFTERBURNER AND SCRUBBER. OBSERVE ALL FEDERAL, STATE AND LOCAL ENVIRONMENTAL REGULATIONS. SECTION 14. - - - - - - - - - - TRANSPORT INFORMATION - - - - - - - - - CONTACT SIGMA CHEMICAL COMPANY FOR TRANSPORTATION INFORMATION. SECTION 15. - - - - - - - - - REGULATORY INFORMATION - - - - - - - - - - REVIEWS, STANDARDS, AND REGULATIONS OEL=MAK IARC CANCER REVIEW:ANIMAL INADEQUATE EVIDENCE IMEMDT 10,99,1976 Page 2 ^ < arc committed to t4e Ivcciii of our Cuttomc. EmMIo7 cc, and SliartkuIdtrt itiTovfh L`i* Science. Teciinolojj and Service. 000429 ___ __ __ __ _ __ 418-018:PAGE C-39 PRODUCT SA 3USU a p r u u o o i sa~ ' #: C8503 NAME: CHOLESTERQIsaini Lous.Missouri63103USA ryjKWA SHEET, Valid 8/2000 - TWi*1*(9C>-S2i-B956i3U)ni-5765 kd 07/25/2000 15:03 Pax800-325-5052(314)771-5757 ____________ ______________________________ email: clg-ld6 iul.com____ ____ ra w rr.B BBvrg.w . w nwaw TManr^rTaTT. m i M a r j : ___tmfimht--n _ 9 V i IARC CANCER REVIEW:GROUP 3 IMSUDL 7,1S1,1987 NOHS 1974: HZD 80865; NIS 8; TNF 1321; NOS 10; TNE 13980 NOES 1983: HZD 80865; NIS 5; TNF 4091; NOS 11; TNE 41411; TFE 30220 EPA GENETOX PROGRAM 1908, NEGATIVE: SHE-CLONAL ASSAY EPA TSCA SECTION 8(B) CHEMICAL INVENTORY EPA TSCA SECTION 8(D) UNPUBLISHED HEALTH/SAFETY STUDIES EPA TSCA TEST SUBMISSION (TSCATS) DATA BASE, DECEMBER 1999 SECTION 16. - - - - - - - - - - OTHER INFORMATION- - - - - - - - - - - - THE ABOVE INFORMATION IS BELIEVED TO BE CORRECT BUT DOES NOT PURPORT TO BE ALL INCLUSIVE AND SHALL BE -USED ONLY AS A GUIDE. SIGMA, ALDRICH, FLUKA SHALL NOT BE HELD LIABLE FOR ANY DAMAGE RESULTING FROM HANDLING OR FROM CONTACT WITH THE ABOVE PRODUCT. SEE REVERSE SIDE OF INVOICE OR PACKING SLIP FOR ADDITIONAL TERMS AND CONDITIONS OF SALE. COPYRIGHT 1999 SIGMA-ALDRICH CO. LICENSE GRANTED TO MAKE UNLIMITED PAPER COPIES FOR INTERNAL USE ONLY Page 3 'V t a r e c o m m i t t e d i a ihc S u c c t J t u/ DU, C m i t n i r i . F m 0 / 0 r cr a n d X h a r t h a l i t n t h r u * t k Life Sennet. Ttchnoiofy and Serv,ct 000430 418-018:PAGE C-40 I/ . ' >J S IG M A -A L D R IC H ________________________ Material Safety Data Sheet Date Printed; 07/25/2000 Date Updated: 11/01/1999 Version 1.0 Section 1 - Product and Company Information Product Name Product Number Brand Company Street Address City, State, Zip, Country Technical Phone: Fax: DL-MEVALONIC ACID LACTONE M4667 Sigma Chemical Sigma-Aldrich 3050 Spruce Street St. Louis, M O . 63103. US 314 771 5765 800 325 5052 Emergency Phone: 414 273 3850 Ext.5996 Section 2 - Composition/lnformation on Ingredient Substance Name DL-MEVALONIC ACID LACTONE Formula Synonyms C6H1003 CAS 674-26-0 SARA 313 No Section 3 - Hazards Identification For additional information on toxicity, please refer to Section 11. Section 4 - First Aid Measures Oral Exposure If swallowed, wash out mouth with water provided person is conscious. Call a physician. Inhalation Exposure If inhaled, remove to fresh air. It not breathing give artificial respiration. If breathing is difficult, give oxygen. Dermal Exposure In case of contact, immediately wash skin with soap and copious amounts of water. Eye Exposure In case of contact, immediately flush eyes with copious amounts of water for at least 15 minutes. Section 5 Fire Fighting Measures Flash Point: 235 *F 113 C Autotgnrtion Temp: N/A Flammability: Extinguishing Media Suitable W ater soray. Carbon dioxide, dry chemical powder, or appropnate foam. N/A 000431 418-018:PAGE C-41 Firefighting Protective Equipment W e a r self-contained breathing apparatus and protective clothing to prevent contact with skin and eyes Section 6 Accidental Release Measures Procedure(s) of Personal Precaution(s) W ear respirator, chemical safety goggles, rubber boots, and heavy rubber gloves. Methods fo r Cleaning Up Absorb on sand or vermicultte and place in dosed containers tor disposal. Ventilate area and wash spill site after material Dickup is complete. Section 7 - Handling and Storage Handling User Exposure Avoid inhalation. Avoid contact with eyes, skin, and clothing. Avoid prolonged or repeated exposure. Storage Suitable Keep tightly closed. Section 8 - Exposure Controls / PPE Engineering Controls Safety shower and eye bath. Mechanical exhaust required. Personal Protective Equipment Respiratory NIOSH/M SHA-approved respirator. Hand Compatible chemical-resistant gloves. Eye Chemical safety goggles. General Hygiene Measures Wash contaminated clothing before reuse. W ash thoroughly after handling. Section 9 - Physicai/Chemical Properties Appearance Color Faintly beige Molecular Weight: 130.15 AMU Prooertv pH BP/BP Range MP/MP Range Freezing Point Vapor Pressure Vapor Density Saturated Vapor Cone. SG /Density Bulk Density Odor Threshold Value N/A 145- 150 -C 24 *c N/A N/A N/A N/A N/A N/A N/A Form Solidified mass or fragments At Temperature or P ra u u m 5 mmHg Sigma Chemical - M 4667 Page 2 Sigma-AJdrieh Corporation Mnww.sigmo-aidnch.com 000432 418-018:PAGE C-42 Volatile% VOC Content Water Content Solvent Content Evaporation Rata Viscoaity Partition Coefficient Decomposition T amp. Flash Point *F Flash Point *C Explosion Limits Autoignition Temp Refractive Index Solubility N/A N/A N/A N/A N/A N/A N/A N/A 235 -F 113 *C N/A N/A 1.473 N/A Section 10 - Stability and Reactivity Stability Conditions of Instability Protect from moisture. Conditions to Avoid Protect from moisture. Materials to Avoid Strong oxidizing agents. Hazardous Decomposition Products Hazardous Decomposition Products Carbon monoxide. Carbon dioxide. Section 11 - Toxicological Information Route of Exposure Skin Contact May cause skin irritation. Eye Contact May cause eye irritation. Inhalation Material may be irritating to mucous membranes and upper respiratory tract. Multiple Routes May be harmlui by inhalation, ingestion, or skin absorption. Signs and Symptoms of Exposure To the best of our knowledge, the chemical, physical, and toxicological properties have not been thoroughly investigated. RTECS Number: Section 12 - Ecological Information Section 13 Disposal Considerations Appropriate Method of Disposal of Substance or Preparation Dissolve or mix the matenal wrth a combustible solvent and bum in a chemical incinerator equipped with an afterburner and scrubber. Observe all federal, state, and local environmental regulations. Section 14 - Transport Information DOT Proper Shipping Name: None Sigma Chemical - M4667 Page 3 Sigme-Aidnch Corporation www.agm-afelhch.com 000433 418-018:PAGE C-43 Non-Haza rdous to r Transport: This substance is considered to be non-hazardous tor transport. IA T A Proper Shipping Name: None Non-Hazardous fo r A ir Transport: Non-hazardous tor air transport. Section 15 - Regulatory Information United States Regulatory Inform ation SARA 313 U sted: No Section 16 - Other Information W arranty The above information is believed to be correct but does not purport to be all inclusive and shall be used only as a guide. SigmaAldrich Inc., shall not be held liable tor any damage resulting from hahdling or from contact with the above product. See reverse siae of invoice or packing slip tor additional terms and conditions ot sale. Copynght 1999 Sigma-Akfnch Co. License granted to make unlimited paper copies tor internal use only. Sigma Chemical - M4667 Page 4 Sigma-AJdrich Corporation w w w .agm e-aM nch.com 000434 418-018:PAGE C-44 ATTACHMENT 3 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE 000435 418-018:PAGE C-45 ATTACHMENT 3 Protocol 418-018 Version: 418-018(16 AUG 00) Page 1 of 5 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE Test Article: Control Articles: Vehicle: PFOS Cholesterol Mevalonic acid 0.5% Tween 80 in R.O. Deionized Water for PFOS and Cholesterol R.O. deionized water for Mevalonic Acid A. Purpose: The purpose of this procedure is to provide a method for the preparation of dosage formulations of PFOS, mevalonic acid, cholesterol and the 0.5% Tween 80 vehicle for oral administration to rats on Primedica Argus Research Laboratories, Inc. Study 418-018. B. General Information: 1. All formulation containers will be labeled and color coded. Each label will specify the protocol number, test article identification, Argus batch number, concentration, dosage level, preparation date, expiration date and storage conditions. 2a. Suspensions of PFOS will be prepared: __ Daily X Weekly __ For____ days of use 2b. Formulations of the control articles will be prepared: X Once Daily (cholesterol) X Twice daily (Mevalonic acid) 2c. The 0.5% Tween 80 Vehicle will be prepared: ___ Daily X Weekly __ For____ days of use 3. Suspensions will be prepared at a final dosage volume of 5 mL/kg. 4. Safety: X Gloves, lab coat, goggles or safety glasses and faceshield X Dust-Mist Respirator __ Half-Face Respirator __ Full-Face Respirator/Positive Pressure Hood __ Tyvek Suit/Apron 5. Dosage formulations of the test article and control articles adjusted for Free base and % Purity. __ Yes X No (Calculations based on 100%) ___ Free Base ___ Purity 000436 418-018:PAGE C-46 ATTACHMENT 3 Protocol 418-018 Version: 418-018116 AUG 00) Page 2 of 5 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol. NOTE: The test article suspensions will be prepared as a serial dilution from the high dosage to the low dosage. Once the final volumes are achieved, stir bars are to be added to the containers; mixing should occur during preparation, aliquotting, sampling and/or dosage administration. C. Preparation of the 0.5% Tween 80 Vehicle: 1. Add 2985 mL of R.O. deionized water to an appropriately labeled container. Heat the water to 5 0 C = 5C, then add 15 mL of Tween 80 and mix until uniform. D. Test Article Suspension Preparation: NOTE: Prior to preparation of the 0.40 mg/mL suspension, precalibrate an appropriately sized container to the desired volume with R.O. deionized water. 1. To prepare the 0.40 mg/mL suspension, weigh the required amount of test article (See TEST ARTICLE PREPARATION CALCULATIONS) into an appropriately sized, labeled, pre-calibrated container. QS ad to the required amount with 0.5% Tween 80 and heat the mixture to 80 C =5C for approximately 30 minutes or until the TA/S dissolves. 2. Once the test article has dissolved, remove the suspension from the water bath, add a magnetic stir bar to the container, place on a magnetic stir plate and mix until the suspension equilibrates to room temperature. (Be sure there is a visible vortex, this will achieve the desired emulsion.) Be sure that the suspension is mixed prior to and during preparation of other dosage groups, aliquotting, dosage and/or sampling. 3. To prepare the 0.32 mg/mL suspension, remove the required amount of 0.40 mg/mL suspension (See TEST ARTICLE PREPARATION CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ad to the final required volume with 0.5% Tween 80 (See TEST ARTICLE PREPARATION CALCULATIONS). Stopper the graduated cylinder and invert several times to ensure proper mixing. Transfer the suspension to an appropriately sized, labeled container. Add a magnetic 000437 418-018:PAGE C-47 ATTACHMENT 3 Protocol 418-018 Version: 418-018(16 AUG 00) Page 3 of 5 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE stir bar to the container, place on a magnetic stir plate and mix prior to and during preparation of other dosage groups, aliquotting, dosage and/or sampling. 4. To prepare the 0.24 mg/mL suspension, remove the required amount of 0.32 mg/mL suspension (See TEST ARTICLE PREPARATION CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ad to the final required volume with 0.5% Tween 80 (See TEST ARTICLE PREPARATION CALCULATIONS). Stopper the graduated cylinder and invert several times to ensure proper mixing. Transfer the suspension to an appropriately sized, labeled container. Add a magnetic stir bar to the container, place on a magnetic stir plate and mix prior to and during preparation of other dosage groups, aliquotting, dosage and/or sampling. 5. To prepare the 0.20 mg/mL suspension, remove the required amount of 0.24 mg/mL suspension (See TEST ARTICLE PREPARATION CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ad to the final required volume with 0.5% Tween 80 (See TEST ARTICLE PREPARATION CALCULATIONS). Stopper the graduated cylinder and invert several times to ensure proper mixing. Transfer the suspension to an appropriately sized, labeled container. Add a magnetic stir bar to the container, place on a magnetic stir plate and mix prior to and during preparation of other dosage groups, aliquotting, dosage and/or sampling. 6. To prepare the 0.16 mg/mL suspension, remove the required amount of 0.20 mg/mL suspension (See TEST ARTICLE PREPARATION CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ad to the final required volume with 0.5% Tween 80 (See TEST ARTICLE PREPARATION CALCULATIONS). Stopper the graduated cylinder and invert several times to ensure proper mixing. Transfer the suspension to an appropriately sized, labeled container. Add a magnetic stir bar to the container, place on a magnetic stir plate and mix prior to and during preparation of other dosage groups, aliquotting, dosage and/or sampling. 7. To prepare the 0.08 mg/mL suspension, remove the required amount of 0.16 mg/mL suspension (See TEST ARTICLE PREPARATION CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ad to the final required volume with 0.5% Tween 80 (See TEST ARTICLE PREPARATION CALCULATIONS). Stopper the graduated 000438 418-018:PAGE C-48 ATTACHMENTS Protocol 418-018 Version: 418-018(16 AUG 001 Page 4 of 5 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE cylinder and invert several times to ensure proper mixing. Transfer the suspension to an appropriately sized, labeled container. Add a magnetic stir bar to the container, place on a magnetic stir plate and mix prior to and during aliquotting, dosage and/or sampling. 8. To prepare the 0 mg/mL suspension, add the required amount of the appropriate vehicle (R.O. deionized water or 0.5% Tween 80) to an appropriately sized, labeled container (See TEST ARTICLE PREPARATION CALCULATIONS). Add a magnetic stir bar to the container, place on a magnetic stir plate and mix prior to and during aliquotting, dosage and/or sampling. E. Preparation of Mevalonic Acid Solution: 1. Weigh the required amount of mevalonic acid into an appropriately labeled container (See TEST ARTICLE/VEHICLE PREPARATION CALCULATIONS). 2. Add approximately 80% of R.O. deionized water to the container. Add a stir bar to the container, place on a magnetic stir plate and agitate until the control article has dissolved completely. 3. Transfer solution to an appropriately sized graduated cylinder and Q.S. to the final desired volume with R.O. deionized water (See TEST ARTICLE/VEHICLE PREPARATION CALCULATIONS). 4. Cover the graduated cylinder with a glass stopper and mix by inversion. Return the solution to the original container. Add a magnetic stir bar to the container, place on a magnetic stir plate and mix prior to and during dosage. F. Preparation of Cholesterol Suspension: NOTE: Prior to preparation of the cholesterol suspension, precalibrate an appropriately sized container to the desired volume with R.O. deionized water. 1. Weigh the required amount of cholesterol into an appropriately sized, labeled container (see TEST ARTICLE/VEHICLE PREPARATION CALCULATIONS). 000439 418-0 L8'.PAGE C-49 ATTACHMENT 3 Protocol 418-018 Version: 418-018(1 6 AUG 00) Page 5 of 5 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE 2. Add a small amount of 0.5% Tween 80 to the container and wet the cholesterol by using a glass rod. QS ad with the 0.5% Tween 80 to the final required volume (see TEST ARTICLE/VEHICLE PREPARATION CALCULATIONS). 3. Add a magnetic stir bar to the vessel, place on a magnetic stir plate and mix until uniform prior to and during dosage administration. Written by: Approved b Clarificatior 000440 418-018:PAGE C-50 ATTACHMENT 4 TISSUE COLLECTION AND PROCESSING 000441 418-018:PAGE C-51 ATTACHMENT 4 Protocol 418-018 Page 1 of 1 Tissue Collection and Processing Sample Storage/Shipping Type Units Aliquots Conditions Shipped to Analvsis Day 21 Gestation Fetuses Blood Pooled/litter 1/2 plasma <-20 PWL Mevalonic acid 1/4 serum <-20 3M PFOS 1/4 serum <-20 AniLytics LDL. HDL. tot chol. trial Liver Pooled/litier 1 liver Gluteraldehvde. RT 3M Possible EM 1/3 all livers <-20 3M PFOS 1/3 all livers <-20 AmLvtics LDL. HDL. tot chol. inal 1/3 all livers Liquid nitrogen. 3M Possible <-70 Biochemical Dav 21 Gestation Dams Blood 4 mL/dam 1/2 plasma <-20 1 PWL Mevalonic acid Oo r r1 VI VI 1/4 serum 3M PFOS 1/4 serum AniLylics LDL. HDL. tot chol. tnglv 1 Li ver each dam right lateral Liquid nitrogen. 3M Possible lobe <-70 Biochemical median lobe <-20 3M PFOS 0.3-0.5 cm section Gluteraldehvde. RT 3M Possible EM remainder o r1 V! Ani Lytics LDL. HDL. tot chol. trialv Day 5 Lactation Dams Milk 100 uL/dam <-20 AmLvtics Total cholesterol Blood 4 mL/dam 1/2 plasma <-20 PWL Mevalonic acid 1/4 serum <-20 3\1 PFOS 1/4 serum <-20 Am Lytics LDL. HDL. tot chol. trial Liver each dam nght lateral Liquid nitrogen. 3M Possible lobe <-70 Biochemical median lobe <-20 3M PFOS 0.3-0.5 cm Gluteraldehvde. RT 3M Possible EM section Remainder <-20 Am Lytics LDL. HDL. tot chol. trialv Heart each dam Gluieraldehyde. RT 3M Possible EM & light microscopv Dav 5 Postpartum Pups co n r1 VI VI Blood Pooled/sex/litter 1/2 plasma PWL Mevalonic acid 1/4 serum <-20 3M PFOS 1/4 serum <-20 AniLytics LDL. HDL. tot chol. trial Liver Pooled/sex/litter 1/3 all livers <-20 3M PFOS 1/3 all livers AniLytics LDL. HDL. tot chol. tngl 1/3 all livers Liquid nitrogen. 3M Possible <-70 Biochemical Hearts 2 male. 2 female, per Gluteraldehvde. RT 3M Possible EM & sex per litter light microscopy PWL = Pnmedica Worcester Laboratories Tot chol = Total cholesterol tngl = Triglyceride RT = Room Temperature EM = Electron Microscopy 000442 418-018:PAGE C-52 O Primedica Argus Research Laboratories. Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215)443-8587 PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 Amendment 1-13 September 2000 1. Storage (page 4 of the protocol): The storage conditions for the Mevalonic Acid should be frozen, rather than room temperature. Reason for Change: This change was made at the request of the Sponsor. 2. Shipping Instructions (page 6 of the protocol): The correct address for Dave Ehresman is the following: Dave Ehresman 3M Corporate Toxicology 3M Center Building 236-0C-148 St. Paul, Minnesota 55144 Telephone: (651)733-5070 Telefax: (651)737-4754 000443 Any revisions made to this finalized amendment must be made by subsequent amendment. Reason for Change: This change corrects the protocol. 418-018:PAGE C-53 Protocol 418-018 Amendment 1 Page 2 Associate Director of Research t uSna C. Lebo, V.M.D. Date Deanna J. LuebKer. M.S. Chairperson, Institutional Animal Care Study Monitor and Use Committee (go Date John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor Any revisions made to this finalized amendment must be made by subsequent amendment. 000444 P rim ed ica 418-018:PAGE C-54 Argus Research Laboratories. Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215)443-8587 PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 Amendment 2 - 22 November 2000 1. Alternate Study Monitor (page 1 of the protocol): The telephone number for John Butenhoff. Ph.D.. DABT, CEH is (651) 733-1962, rather than (651) 737-1962. Reason for Change: This change corrects the protocol. 2. Sample Analysis and Shipping Instructions (page 14 of the protocol): The following paragraphs will replace the paragraph which states that serum and liver samples will be analyzed for LDL, HDL, total cholesterol and triglycerides: The remaining liver samples will be analyzed for LDL, HDL, total cholesterol and triglycerides. Serum samples collected from all dams and pups will be analyzed for total cholesterol, glucose, total and free T3. T4and TSH levels, LDL, HDL and triglycerides. The first priority will be analysis for total cholesterol and glucose. The second priority will be analysis for total and free T 3. T4 and TSH levels. The third priority will be analysis for LDL. HDL and triglycerides. 000445 Any revisions to this finalized amendment must be made by subsequent amendment 418-018:PAGE C-55 r I CJ The liver and serum samples will be shipped to: Protocol 418-018 Amendment Page Dr. Saroj R. Das Ani Lytics, Inc. 200 Gerard Street, Suite 2000 Gaithersburg, Maryland 20877 Telephone: (301)921-0168 Telefax: (301)977-0433 Reason for Change: This change was made at the request of the Sponsor in order to include additional tissue samples and to prioritize the use of samples for analyses. 3. Scheduled Sacrifice - FI Generation Puds (page 20 of the protocol): [Effective Date: 3 November 2000] For Replicate 1, blood samples will be pooled per litter, samples from male and female pups combined, rather than pooled per sex. Samples will be aliquoted as follows, rather than as described in paragraph 3. Approximately 1mL of blood will be transferred into serum separator tubes; any remaining blood will be transferred into EDTA-coated tubes. The samples will be spun in a centrifuge and the resulting serum will be divided into two aliquots. The first aliquot (approximately 200 p.L) will be used for PFOS analysis. The second aliquot (approximately 300 pL of serum) will be used for analysis of total cholesterol, glucose, total and free T 3 .T 4 and TSH levels and LDL, HDL and triglyceride levels, according to the priorities described in item 1, above. Any resulting plasma will be transferred into one aliquot. Resulting serum will be prioritized as follows: clinical chemistries first (according to the priorities described in item 2, above) and then PFOS samples. All samples will be immediately frozen on dry ice and maintained frozen (:s-70C) until shipment for analysis. Reason for Change: This change was made at the request of the Sponsor in order to prioritize the use of samples for analyses. 4. Scheduled Sacrifice - FI Generation Pups (page 20 of the protocol): [Effective Date: 17 November 2000] For Replicate 2, blood samples will be pooled (two litters per sample within each dosage group). Samples will be aliquoted as follows, rather than as described in paragraph 3. Approximately 0.3 mL of blood will be transferred into EDTA-coated tubes. The remaining blood will be transferred into serum separator tubes. The samples will be spun in a centrifuge and the resulting serum will be divided into two aliquots. Resulting plasma will be transferred into one aliquot. The serum will be prioritized as follows: clinical chemistries first (according 00044$ Any revisions to this finalized amendment must be made by subsequent amendment. 418-018:PAGE C-56 Protocol 418-018 Amendment 2 Page 3 to the priorities described in item 2, above) and then PFOS samples. All samples will be frozen on dry ice and maintained frozen (<-70C) until shipment for analysis. Reason for Change: This change was made at the request of the Sponsor in order to prioritize the use of samples for analyses. 5. Scheduled Sacrifice - FI Generation Pups (page 21 of the protocol): [Effective Date: 3 November 2000] Thyroids will be excised from each pup, pooled (by sex per litter). Thyroids will be retained in neutral buffered 10% formalin for possible future evaluation. Reason for Change: This change was made at the request of the Sponsor. Sorge. Dearlove, Ph.D., DABT Associate Director of Research Date R&ond G. Yofc/mD., DABT Date Associate Director of Research Study Director Chairperson, Institutional Animal Care and Use Committee La lCa.0 4 2 Deanna J. Luebker, M.S. Study Monitor Date John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor 000447 Any revisions to this finalized amendment must be made by subsequent amendment 418-018:PAGE C-57 O Primedica Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215)443-8587 PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 Amendment 3-11 January 2001 1. Control Articles (page 3 of the protocol): The lot number for mevalonic acid is lot number 070K26603, rather than 070K2603. Reason for Change: This change corrects a typographical error. 2. Control Articles (page 3 of the protocol): For mevalonic acid, lot number 080K2618, expiration date September 2004, was used in addition to lot number 070K26603. Reason for Change: Additional mevalonic acid was required for the preparation of the mevalonic acid control and PFOS plus mevalonic acid formulations. 3. Sample Analysis and Shipping Instructions (page 14 of the protocol): Plasma mevalonic acid levels will be measured for the following dosage groups: Vehicle Control (Group 1), Mevalonic Acid Control (Group 2), 1.6 mg/kg PFOS + Mevalonic Acid (Group 3), 2 mg/kg PFOS + Mevalonic Acid (Group 4), 1.2 mg/kg PFOS (Group II), 1.6 mg/kg PFOS (Group 12) and 2 mg/kg PFOS (Group 13), rather than all dosage groups. Mevalonic acid levels may be measured in the lower PFOS dosage groups after evaluation of the results of the above analyses. Remaining samples will be retained at Primedica Worcester Laboratories or the Testing Facility until notification from the Sponsor for disposition. 000448 Any revisions made to this finalized amendment must be made by subsequent amendment. 418-018:PAGE C-58 Reason for Change: Protocol 418-018 Amendment 3 Page 2 This change was made at the request of the Sponsor. 4. Sample Analysis and Shipping Instructions (page 1 of Amendment 2 to the protocol): Total and free T3, T4and TSH levels will be measured for the following dosage groups: Vehicle Control (Group 1), 1.2 mg/kg PFOS (Group 11), 1.6 mg/kg PFOS (Group 12) and 2 mg/kg PFOS (Group 13), rather than all dosage groups. Total and free T 3 , T4 and TSH levels may be measured in the lower PFOS dosage groups after evaluation of the results of the above analyses. Reason for Change: This change was made at the request of the Sponsor. Study Director Chairperson, Institutional Animal Care and Use Committee Study Monitor John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor 000449 Any revisions made to this finalized amendment must be made by subsequent amendment. 418-018.-PA G EC -59 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215) 443-8587 PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 Amendment 4 - 1 8 April 2001 1. F 1 Generation P u p s (page 10 of the protocol): [Effective Date: 21 August 2000] Day 1 of lactation (postpartum) is defined as the day of birth and is also the first day on which all of the pups in a litter are weighed. The pups will not be individually weighed; pooled litter weights will be recorded. Reason for Change: This change was made to be consistent with Tests, Analyses and Measurements FI Generation, Body Weights. 2. Scheduled Sacrifice - FI Generation P u p s (page 20, 21, page 1 of ATTACHMENT 4 and item 5 of Amendment 2 to the protocol): Histopathology will be performed on the heart and thyroid of one male and one female FI generation pup from each of four litters from the Vehicle Control group (Group 1) and 2 mg/kg PFOS group (Group 13). If any difference is found, histopathology will be performed on the heart and thyroid of one male and one female FI generation pup from each of four litters, when possible, from the 0.4 and 1.6 mg/kg PFOS dosage groups (Groups 8 and 12, respectively). Tissues will be shipped (ambient conditions) for evaluation to: W. Ray Brown, D.V.M., Ph.D., ACVP Veterinary Pathologist Research Pathology Services, Inc. 438 E. Butler Avenue New Britain, Pennsylvania 18901 Telephone: (215)345-7070 Telefax: (215) 345-4326 000450 Any revisions made to this finalized amendment must be made by subsequent amendment. Reason for Change: This change was made at the request of the Sponsor. 418-018:PAGE C-60 Protocol 418-018 Amendment 4 Page 2 T George . Dearlove, Ph.D., DABT Associate Director of Research ji- e e - * / Date Date Associate Director oNLesearch and Study Director (g|iin<^ fn t iiT Y ii leresa H. Woodird, D.V.M. Date Chairperson, Institutional Animal Care and Use Committee Deanna J. Luebi r, M.S. Study Monitor Date John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor 000451 Any revisions made to this finalized amendment must be made by subsequent amendment 418-018:PAGE C-61 905 Sheehy Drive. BWg- A Horsham, PA 19044 Telephone: (2 IS) 443-8710 Telefax: (215) 443-8587 ARGUS RESEARCH Charles River Laboratories Discovery and Development Services PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 Amendment 5 - 4 February 2002 1. Shipping Instructions (page 6 of the protocol): [Effective Date: 29 January 2002] Homogeneity and concentration samples shipped to Dave Ehresman at 3M Corporate Toxicology for analysis will be reshipped (frozen on dry ice) to: Emily R. Decker (Principal Investigator) Exygen Research 3058 Research Drive State College, Pennsylvania 16801 Telephone: (814)272-1039 Telefax: (814)272-1019 Email: edecker@centrelab.com The analyses will be subcontracted to Exygen Research by the Sponsor and the Quality Assurance Unit for Exygen Research will conduct critical phase inspections and audit the respective results and reports according to the Standard Operating Procedures of that facility. Such critical phase inspection reports and audit reports will be submitted by that facility to the Study Director, Raymond G. York. The date of the inspections and report submissions will be incorporated into a QAU statement generated by Exygen Research for inclusion in the final report for protocol 418-018. Any revisions to this finalized amendment must be made by subsequent amendment 000452 Reason for Change: This change was made at the request of the Sponsor. 418-018:PAGE C-62 Protocol 418-018 Amendment 5 Page 2 orge Dearlove, Ph.D., DABT Associate Director of Research ond G. h.D., DABT Date Associate Di: f Research and Study Director // {.U iT yJa^J Hpi Theresa H. Woodard, D.V.M. Date Chairperson, Institutional Animal Care and Use Committee Deanna J. Luebner, M.S. Study Monitor Date John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor Any revisions to this finalized amendment must be made by subsequent amendment. 000453 418-018:PAGE C-63 905 Shcehy Drive, Bldg. A Horsham, PA 19044 Telephone: (U S ) 443-8710 Telefax: (215) 443-8587 ARGUS RESEARCH Charles River Laboratories Discovery a n d Development Services PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 Amendment 6 - 9 October 2002 1. Shipping Instructions (page 6 of the protocol and Amendment 5, item 1): [Effective Date: 14 February 2002] The samples shipped to Exygen Research will be analyzed using the LC/MS/MS conditions found in the attached pages. Reason for Change: This change was made at the request of Exygen Research to clarify the analytical methodology to be used for the analysis of the samples. 2. Shipping Instructions (page 6 of the protocol and Amendment 5, item 1): [Effective Date: 16 September 2002] The e-mail address for Emily R. Decker is emily.decker@exygen.com. Any revisions to this finalized amendment must be made by subsequent amendment 000454 418-018:PAGE C-64 Reason for Change: Protocol 418-018 Amendment 6 Page 2 This change was made because the e-mail address was changed. a f vm i Veor E. Dearlove, Ph.D., DABT Associate Director of Research Date Rayhrfnki G. York, AssocWe Director and Study Director ABT Date Farch C . .L U - ,, Dena C. Lebo, V.M.D. Member, Institutional Animal Care and Use Committee Date Deanna J. Luebist, M.S Study Monitor L m ijjfo^ s Date John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor Any revisions to this finalized amendment must be made by subsequent amendment 000455 418-018:PAGE C-65 LC/MS/MS System and Operating Conditions (Turbolonspray) Mass Spec: PE SCIEX API 3000 Biomolecular Mass Analyzer Interface: SCIEX Turboion Spray Liquid Introduction Interface Harvard infusion pump Computer Dell UltraScan PI 110 Software: PE SciexAnalyst 1.1 Windows NT HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven HPLC Column:Genesis Cg (Jones Chromatography), 2.1 mm x 50 mm, 4p Column Temp.: 35 C Injection Vol.: 10 pL Mobile Phase (A): 2 mM Ammonium Acetate m ASTM type I water Mobile Phase (B): Methanol Flow Rate: 0.3 mL/min. Time % A % B 0 60 40 1.0 0 100 7.0 0 100 7.5 60 40 11.0 60 40 It may be necessary to adjust the HPLC gradient in order to optimize instrument performance. Columns with different dimensions (e.g., 2.1 mm x 30 mm) and columns from different manufacturers (Keystone Betasil Cig etc.) can be used, provided equivalent chromatography is obtained. Ions monitored: Analyte PFOS Mode negative Transition Monitored 499 -> 99 Approximate Retention Time (min) 4.40 000456 ^^^3058 Research Drive .# ^ S ta te College, PA16801, USA 800-2813219 i R 814.272.1019 exygen.com TM RESEARCH Precise Research. Proven Results. 418-018:PAGE C-66 On a day-to-day basis, the retention times may vary slightly depending on the batch of mobile phase, etc. Example Tune File Parameters The following values are provided as an example. Actual values may vary from instrument to instrument. Also, these values may be changed from time to time in order to optimize for greatest sensitivity. Once the instrument is tuned, the optimized parameters are saved as a "tune file". This tune file is then used during routine analysis. The tuning procedure may be repeated as necessary to ensure optimal sensitivity. Tune File Parameters Controls IS-Iospray DP-Declustering Potential FP-Focusing Potential EP-Entrance Potential CE-Collision Energy CXP-Collision Cell Exit Potential DF-Deflector CEM-Channel Electron Multiplier Set -4200.0 -51.0 -230.0 10.0 -70 -5 300.0 2800.0 Gas Flows Nebulizer Gas Curtain Gas Collision Gas TTS Temperature Set 12 13 4 350C Calibration Procedures a. Inject the same volume (between 10 to 20 pJL) of each calibration standard (prepared in MeOH) into the LC/MS/MS. b. Use linear, I/x weighted standard curves for quantification. Linear standard curves are generated for each set by linear regression using the appropriate software system. Any calibration standards falling outside 30%, based on its calculated concentration, may be excluded from the calibration curve. However, the total number of calibration standards that may be excluded must not exceed 20% of the total number of standards injected, c. The correlation coefficient (r) for calibration curves generated must be >0.9925 (r2>0.985). If calibration resi fell outside these 3058 Research Drive i State College, PA16801, USA 000457 : 800-281.3219 F: 814.272.1019 exygen.com TM RESEARCH Precise Research. Proven Results. 418-018:PAGE C-67 limits, then appropriate steps should be taken to adjust instrument operation, and the relevant set of samples must be reanalyzed. Sample Analysis Inject the same volume used for the calibration standards (between 10 to 20 pL) of each sample, fortification, control, etc. into the LC/MS/MS. Each sample must be analyzed in duplicate. a. Standards corresponding to at least six concentrations must be included in an analytical set. b. Inject an entire set of standards (six) at the beginning of the run and inject standards interspersed about every 5-10 samples. All sample injections must be bracketed by standard injections. c. The concentration of each sample, fortification, control, etc. is determined from the standard curve based on the peak area of the analyte in all standards injected during a set. The standard responses must bracket responses of the residue found in the sample set. If necessary, dilute the samples and re-analyze to give a response within the standard curve range. 000458 exygen.com APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 000459 418-018: PAGE D -l DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 1. Rats were ordered to weigh 175 g to 200 g at receipt, rather than 175 g to 225 g at receipt. This deviation did not adversely affect the outcome or interpretation of the study because the rats were a sufficient weight to go on study. 2. The following dosage administrations were performed from two minutes early to 60 minutes late. Dosage Number Dosage Administered Group Identification Date Day of Study" Early or Late Rat Number 1 Vehicle 15 SEP 00 DS 2 2 11501 - 11514 Control 22 SEP 00 DS 9 3 11501 - 11514 10 OCT 00 DS 43, DG 0 3 10901 - 10914 14 OCT 00 DS 47, DGs 1- 4 3 10901 - 10914 14 OCT 00 DS 31 3 11501 - 11514 20 OCT 00 DS 9 2 10901 24 OCT 00 DGs 2, 11 - 14 2 10901 - 10914 10 NOV 00 DG 19 2 10913 2 Mevalonic 15 SEP 00 DS 2 2 11515, 11517 Acid Control 22 SEP 00 DS 9 3 11515 - 11528 28 SEP 00 DS 15 2 11526- 11528 10 OCT 00 DS 43, DG 0 3 10915 - 10928 14 OCT 00 DS 47, DGs 1- 4 3 10915 - 10928 14 OCT 00 DS 31 3 11515 - 11528 24 OCT 00 DGs 3, 9, 11 - 14 2 10915 - 10928 10 NOV 00 DG 20 2 10927 3 1.6 mg/kg 16 SEP 00 DS 19 2 10930 PFOS + 22 SEP 00 DS 9 3 11529- 11543 Mevalonic 28 SEP 00 DS 15 2 11529- 11543 Acid 10 OCT 00 DS 43, DG 0 3 10929 - 10942 14 OCT 00 DGs 2 - 4 3 10929 - 10942 14 OCT 00 DS 31 3 11529- 11543 24 OCT 00 DGs 12, 14 2 10929- 10932, 10934 4 2 mg/kg PFOS 16 SEP 00 DS 19 2 10943 - 10945 + Mevalonic 22 SEP 00 DS 9 3 11544- 11557 Acid 28 SEP 00 DS 15 2 11544- 11557 10 OCT 00 DS 43, DG 0 3 10943 - 10956 14 OCT 00 DS 47, DGs 0 - 4 3 10943 - 10956 14 OCT 00 DS 31 3 11544- 11557 10 NOV 00 DG 20 2 10952 5 Cholesterol 28 SEP 00 DS 15 2 11558 - 11571 Control 10 NOV 00 DG 18 2 10967 a. DS is used as an abbreviation for day of study. DG is used as an abbreviation for day of (presumed) gestation. DL is used as an abbreviation for day of lactation. 000460 418-018: PAGE D-2 Dosage Group 6 7 Identification 1.6 mg/kg PFOS + Cholesterol 2 mg/kg PFOS + Cholesterol Date 21 SEP 00 28 SEP 00 07 OCT 00 11 OCT 00 21 OCT 00 01 NOV 00 17 NOV 00 02 SEP 00 28 SEP 00 11 OCT 00 12 OCT 00 Day of Study DS 8 DS 15 DS 24 DS 44 DG 8 DG 20, DL 2 DG 21, DL 2 DS 5 DS 15 DS 44, DG 0 DS 45, DG 0 21 OCT 00 DGs 9, 11 01 NOV 00 01 NOV 00 02 NOV 00 17 NOV 00 DG 21 DG 6 DS 50, DGs 4, 5, / DL 1 Dosage Number Administered Early or Late 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 Rat Number 11583 - 11585 11572- 11585 11572 10984 10983 10972, 10973 11584, 11585 10985 11586- 11595 10994 - 10998 10990, 10992, 10994 10985 - 10989, 10991 10997, 10998 11591 11591 - 11599 2 11586 These deviations did not adversely affect the outcome or interpretation of the study because the time deviated was small and the deviation occurred on no more than four days of the approximately 60 to 80 day dosage period for any rat. 3. At least one event of dosage administration and/or postdosage observation was not recorded for the following rats. Dosage Group 1 2 Identification Vehicle Control Mevalonic Acid Control Date 28 SEP 00 28 SEP 00 Day of Study DS 15 DS 15 Rat Number 11502- 11512 11513-11524 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 4. Dosage administration was not performed for the following rats. Dosage Group 7 10 Identification 2 mg/kg PFOS + Cholesterol 1mg/kg PFOS Dosage Not Administered Cholesterol PFOS Date 28 SEP 00 01 OCT 00 Day of Study DS 15 DS 18 Rat Number 11596 11623 These deviations did not adversely affect the outcome or interpretation of the study because it was a single occurrence for each rat. 5. On 1 October 2000, DS 18, the following rats in Group 3, 1.6 mg/kg PFOS + Mevalonic Acid, were administered the incorrect volume of test article. 000461 418-018: PAGE D-3 Rat Number 11532 11537 11539 11541 Target Volume (mL) 1.2 1.2 1.0 1.1 Actual Volume (mL) 1.3 1.4 1.3 1.3 These deviations did not adversely affect the outcome or interpretation of the study because only four rats were affected and the additional volume was small (0.1 to 0.3 mL). 6. On 10 October 2000, DS 43 or DG 0, rats 10943 through 10956 in Group 4, 2 mg/kg PFOS + Mevalonic Acid, were administered the 0.32 mg/mL concentration of PFOS, rather than the 0.40 mg/mL at 5 mL/kg. Approximately 20 minutes later the rats were administered the 0.08 mg/mL concentration of PFOS at 5 mL/kg. These deviations did not adversely affect the outcome or interpretation of the study because the total correct dosage was administered to all of the rats. 7. On 20 November 2000, DL 3, rat 11589 in the 2 mg/kg PFOS + Cholesterol dosage group (Group 7) received 1.9 mL of test article rather than 1.5 mL. This deviation did not adversely affect the outcome or interpretation of the study because the amount of extra volume received was small and this was a single event. 8. Clinical observations, body weights and dosage administration were not recorded for the following rats. Dosage Group 12 13 Identification 1.6 mg/kg PFOS 2 mg/kg PFOS Date 01 NOV 00 07 NOV 00 07 NOV 00 Day of Study DS 65 DG 19 DG 19 DG 21 DG 12 DG 9 DG 12 DG 0 DG 9 DG 10 DG 12 DG 12 DG 10 DG 11 DG 11 DG 10 DG 11 DG 10 Rat Number 11039 11041 11042 11043 11647 11648 11649 11650 11651 11652 11653 11654 11655 11656 11657 11658 11659 11660 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate these parameters. 000462 418-018: PAGE D-4 9. Data for the following rats were not collected due to a computer filing error. Dosage Group 1 3 4 5 Identification Vehicle Control 1.6 mg/kg PFOS + Mevalonic Acid 2 mg/kg PFOS + Mevalonic Acid Cholesterol Control Date 20 SEP 00 20 SEP 00 20 SEP 00 20 OCT 00 Day of Study DS 7 DS 7 DS 7 DG 10, 7 Rat Number 11501 - 11512 11538 - 11543 11544- 11548 10965, 10966 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available for evaluation of this parameter. 10. The feed left and/or feed fed values were not recorded for the following rats. Dosage Group 3 4 Identification 1.6 mg/kg PFOS + Mevalonic Acid 2 mg/kg PFOS + Mevalonic Acid Date 02 NOV 00 21 SEP 00 Day of Study DG 21 DS 8 Rat Number 10939 11553 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 11. Maternal behavior was not recorded for the following rats. Dosage Group 1 2 5 6 Identification Vehicle Control Mevalonic Acid Control Cholesterol Control 1.6 mg/kg PFOS + Mevalonic Acid Date 04 NOV 00 04 NOV 00 19 NOV 00 22 NOV 00 03 NOV 00 Day of Study DL 1 DL 1 DL 5 DL 5 DL 1 Rat Number 10914 10923,10924 10967 11567 10977 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 000463 418-018: PAGE D-5 12. The terminal blood collection data (time collected, volume collected, performed by and date) was not recorded for the following rats. Dosage Group 3 4 6 8 10 Identification 1.6 mg/kg PFOS + Mevalonic Acid 2 mg/kg PFOS + Mevalonic Acid 1.6 mg/kg PFOS + Cholesterol 0.4 mg/kg PFOS 1 mg/kg PFOS Date 02 NOV 00 02 NOV 00 03 NOV 00 22 NOV 00 22 NOV 00 Day of Study DG 21 DG 21 DG 21 DL 5 DL 5 Rat Number 10931 10949 10980 11601 11629 These deviations did not adversely affect the outcome or interpretation of the study because the blood was collected and available for evaluation. 13. On 31 October 2000, DG 21, the maternal liver weight for rat 10932 in the 1.6 mg/kg PFOS + Mevalonic Acid dosage group (Group 3) was not recorded. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 14. The pooled litter weights were not recorded for the pups of the following dams. Dosage Group 5 8 Identification Cholesterol Control 0.4 mg/kg PFOS Date 22 NOV 00 22 NOV 00 Day of Study DL 4 DL 5 Rat Number 11608 11569 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 15. On 19 November 2000, DL 3, the necropsy observations for rat 11554 in the 2 mg/kg/day PFOS + Mevalonic Acid dosage group (Group 4) were not recorded. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 000464 418-018: PAGE D-6 The weight of the pooled liver samples was not recorded for the male pups of the following dams. Dosage Group 1 4 Identification Vehicle Control 2 mg/kg PFOS + Mevalonic Acid Date 25 NOV 00 25 NOV 00 Day of Study DL 5 DL 5 Rat Number 11504 11547 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. All deviations are documented in the raw data. 000465 APPENDIX E CERTIFICATE OF ANALYSIS 00046S 418-018'.PAGE E-l Certificate O f Analysis FC-95, Lot 217 March 9,2000 Richaid M. Payfer This sample was analyzed using LC/MS, *H-NMR, l9F-NMR, and elemental analyses techniques. The re: ofthese tests show the sample to contain the following weight percent composition: CCajFF'iSSO0a3TiCr CsFnSOjTC C&ZSOsK' CgFj' s0 3 lc^ c K S o^tT C|oF2iS 03'K r -------------SFjC#FjtS03ir 0.04% 0.83 % 1.38 % 1.30% 3.71 % U9 % 90.49 % 0.49 % oU % 0.04 % 033% 035 % Additionally, the isomer distribution of the sample was determined using l9F-NMR techniques and found tc contain the M ow ing mole percent composition: C F j(C F ,V S O ,-K ' (Normal chain, where x is mainly 7) c f 3(CF2VCF(c f 3h c f 2V scv k t (Internal monomethyl branch, where x+y is mainly 5, and x * 0, y # 0) (C FjhC F-iC FzV SO s'K ' (Isopropyl branch, where x is mainly 5) C J F ^ i- C F ^ ) - SOs'KT (Alpha branch, where x is mainly 6) (CF3)3C-(CF2V S 0 3 'ir (trbutyl branch, where x is mainly 4) CF3-CCF2>*-C(CF3M C F 2)y- SOs' K? (Internal gem-dimethyl branch, where x+y is mainly 4, and x * 0) 70.5% 17.1% 1.6% 0.2% 02% 000467 418-018:PAGE E-2 SIG M A 3050 Spruce Street Saint Louis, Missouri 63103 USA Telephone 800-521-8956 (314) 771-5765 Fa* 800-325-5052 (314) 771-5757 email: sig-aldOsial.com www.sigma-aidrich.com CERTIFICATE OF ANALYSIS DL-MEVALONIC ACID LACTONE PRODUCT NO: M4667 LOT NO: 070K2603 FORMULA: CgH10O3 FORMULA WEIGHT: 130.1 CAS NO: 674-26-0 STORE IN A FREEZER TEST SPECIFICATION RESULTS APPEARANCE WHITE WITH A DARK YELLOW CAST SEMISOLID CONFORMS SOLUBILITY CLEAR TO VERY SLIGHTLY HAZY FAINT YELLOW SOLUTION AT 400 MG PLUS 4 ML OF ETHANOL CLEAR FAINT YELLOW IDENTITY CONSISTENT WITH STRUCTURE BY IR OR NMR CONFORMS BY NMR PURITY BY TITRATION APPROX. 97% 99.2% QC ACCEPTANCE DATE JULY 2000 DAVID FELDKER MANAGER, ANALYTICAL SERVICES 191/200007#1/YXB1 DOCUMENT DATE: 07/26/00 SIGMA-ALDRICH, INC. WARRANTS THAT ITS PROOUCTS CONFORM TO THE INFORMATION CONTAINED IN THIS AND OTHER SIGMA-ALDRICH PUBLICATIONS. PURCHASER MUST DETERMINE THE SUITABILITY OF THE PROOUCT(S) FOR THEIR PARTICULAR USE. ADDITIONAL TERMS AND CONDITIONS MAY APPLY. PLEASE SEE REVERSE SIDE OF THE INVOICE OR PACKING SLIP. SIGMA BRAND PROOUCTS ARE SOLD EXCLUSIVELY THROUGH SIGMA-ALDRICH, INC. 000468 We ar e c o m m i i i e d to t he S u c c e s s o f o u r C u s t o m e r s , E m p l o y e e s a nd Shareholders t/iroufli Life Science . T e c h n o J o o and Service. APPENDIX F HOMOGENEITY AND CONCENTRATION ANALYSIS REPORT 000469 418-018:PAGE F-l STUDY TITLE One Generation Reproduction Study of PFOS - Mevalonic Acid/Cholesterol Challenge and NOEL Investigation in Rats DATA REQUIREMENTS Analytical Method Requirements STUDY DIRECTOR Raymond G. York, Ph.D., DABT ANALYTICAL PORTION COMPLETED ON October 30, 2002 PERFORMING LABORATORY Exygen Research 3058 Research Drive State College, PA 16801 Phone: 814-272-1039 STUDY SPONSOR 3M Corporate Toxicology St. Paul, MN 55144-100 PROJECT Study Protocol Number: 418-018 Exygen Study Number: 023-069 Total Pages: 21 000470 418-018:PAGE F-2 Exygen Study No.: 023-069 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exygen Study Number 023-069, entitled "One Generation Reproduction Study of PFOS Mevalonic Acid/Cholesterol Challenge and NOEL Investigation in Rats," conducted for 3M Corporate Toxicology, was performed in compliance with U.S. Food and Drug Administration Good Laboratory Practice Regulations by Exygen Research. imny R. Docker Principal Investigator Exygen Research Date / O .f Deanna J. Luebker,\jW.S. Sponsor Representative 3M Corporate Toxicology Exygen Research f t !CT7 /rZ W O ^ Date 000471 Page 2 of 21 418-018:PAGE F-3 Exygen Study No.: 023-069 QUALITY ASSURANCE STATEMENT Exygen Research's Quality Assurance Unit reviewed Exygen Study Number 023-069, entitled, "One Generation Reproduction Study of PFOS - Mevalonic Acid/Cholesterol Challenge and NOEL Investigation in Rats." All reviewed phases were inspected for conduct according to Exygen Research's Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Study Director and to management. Phase 1. Protocol Review 2. Extraction, Fortification 3. Raw Data Review 4. Draft Report Review 5. Final Report Review Date Inspected Date Reported to Date Reported to Exygen Study Director and Management Sponsor Management 2/14/02 2/25/02 4/11/02 2/15/02 2/25/02 4/11/02 4/3/02 9/9/02 8/ 1/02 9/13/02 8/ 8/02 9/18/02 10/ 8/02 10/30/02 10/30/02 Date Exygen Research 000472 Page 3 of 21 418-018:PAGE F-4 Exygen Study No.: 023-069 CERTIFICATION OF AUTHENTICITY This report, for Exygen Study Number 023-069, is a true and complete representation of the raw data for the study. Submitted by: Exygen Research 3058 Research Drive State College, PA 16801 (814)272-1039 Principal Investigator, Exygen: V Emily R. Dicker Scientist Exygen Research _________________ tdj&kL Date Exygen Research Facility Management: 3o ^-T-oz^ Date Study Director, Argus Research: Sponsor Representative, 3M: 3M Corporate Toxicology Exygen Research / y - A J o v -QZ_, Date _i ( /a i A ra , Date 000473 Page 4 of 21 418-018:PAGE F-5 Exygen Study No.: 023-069 STUDY IDENTIFICATION One Generation Reproduction Study of PFOS - Mevalonic Acid/Cholesterol Challenge and NOEL Investigation in Rats STUDY PROTOCOL NUMBER: 418-018 EXYGEN STUDY NUMBER: 023-069 TYPE OF STUDY: Analytical SAMPLE MATRIX: Dosing Solutions TEST SUBSTANCE: Perfluorooctanesulfonate (PFOS) SPONSOR: 3M Corporate Toxicology St. Paul, MN 55144-100 STUDY DIRECTOR: Raymond G. York, Ph.D., DABT Argus Research 905 Sheehy Drive, Building A Horsham, PA 19044-1297 SPONSOR REPRESENTATIVE: Deanna J. Luebker, M.S 3M Corporate Toxicology 3M Center, Building 220-2E-02 St. Paul, MN 55144-1000 TESTING FACILITY: Exygen Research 3058 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: 08/21/00 Analytical Start Date: 02/14/02 Analytical Termination Date: 02/15/02 Analytical Report Completion Date: xx/xx/xx Exygen Research 000474 Page 5 of 21 418-018:PAGE F-6 Exygen Study No.: 023-069 PROJECT PERSONNEL The Study Director for this project was Raymond G. York, Ph.D., DABT at Argus Research. The following personnel from Exygen Research were associated with various phases of the study: Name Emily Decker Karen Risha Rickey Keller Title Scientist/Principal Investigator Scientist/Principal Investigator Sample Custodian Exygen Research 000475 Page 6 of 21 418-018:PAGE F-7 Exygen Study No.: 023-069 TABLE OF CONTENTS Page TITLE PAGE...................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT.............................2 QUALITY ASSURANCE STATEMENT......................................................................... 3 CERTIFICATION OF AUTHENTICITY.......................................................................... 4 STUDY IDENTIFICATION.............................................................................................. 5 PROJECT PERSONNEL................................................................................................... 6 TABLE OF CONTENTS.......................................................................... !...................... 7 LIST OF TABLES.............................................................................................................. 8 LIST OF FIGURES............................................................................................................ 8 LIST OF APPENDICES.................................................................................................... 8 1.0 SUMMARY................................................................................................................9 2.0 OBJECTIVE........................................................................................... 9 3.0 INTRODUCTION.......................................................................................................9 4.0 TEST SYSTEM...........................................................................................................9 5.0 REFERENCE MATERIAL......................................................................................... 9 6.0 DESCRIPTION OF ANALYTICAL METHOD....................................................... 10 6.1 Extraction Procedure............................................................................................... 10 6.2 Preparation of Standards and Fortification Solutions.............................................. 10 6.3 Chromatography...................................................................................................... 11 6.4 Instrument Sensitivity............................................................................................. 11 6.5 Description of Instrument and Operating Conditions.............................................. 11 6.6 Quantitation and Example Calculation....................................................................12 7.0 EXPERIMENTAL DESIGN..................................................................................... 14 8.0 RESULTS.................................................................................................................. 14 9.0 CONCLUSIONS....................................................................................................... 14 10.0 RETENTION OF DATA AND SAMPLES............................................................. 14 Exygen Research 000476 Page 7 of 21 418-018:PAGE F-8 Exygen Study No.: 023-069 LIST OF TABLES Table I. Summary of PFOS in Dosing Solution Samples Page ....17 Figure 1. LIST OF FIGURES Page Typical Calibration Curve for PFOS.............................................................19 Figure 2. Chromatogram Representing a 0.5 ng/mL Calibration Standard for PFOS... 20 Figure 3. Chromatogram Representing Dosing Solution Sample (Exygen ID: 0200283, Set: 021402A).............................................................................................. 21 LIST OF APPENDICES Page Appendix A Study Protocol 418-018 (Exygen Study No. 023-069) and Amendments.. 22 Exygen Research 000477 Page 8 of 21 418-018 .PAGE F-9 Exygen Study No.: 023-069 1.0 SUMMARY Exygen Research analyzed dosing solution samples for the determination of perfluorooctanesulfonate (PFOS) according to protocol 418-018 (Appendix A). Recoveries for PFOS in the dosing solutions ranged from 83% to 105% when compared to the assigned concentration. 2.0 OBJECTIVE The objective of this study was to determine levels of perfluorooctanesulfonate (PFOS), dosing solution samples using the analytical conditions described in Protocol Amendment #6. 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFOS in the dosing solution samples using the analytical conditions described in Protocol Amendment #6. The study was initiated on August 21, 2000, when the study director signed protocol number 418-018. The analytical start date was February 14, 2002, and the analytical termination date was February 15, 2002. 4.0 TEST SYSTEM Twenty-seven dosing solution samples were received frozen on dry ice at Exygen Research on January 31, 2002 and logged in by Exygen personnel and placed in frozen storage. Sample log-in and chain of custody information can be found in the raw data package associated with this study. Storage records will be kept at Exygen Research and a true copy of the storage records is available upon request. 5.0 REFERENCE MATERIAL The analytical standard PFOS was received at Exygen on May 17, 2000 from 3M Environmental Technology and Safety Services. Exygen Research 000478 Page 9 of 21 418-018:PAGE F-10 Exygen Study No.: 023-069 The available information for the reference material is listed below. The reference material PFOS was stored frozen. Compound Exygen Control No. PFOS SP248 TCR No. SD009 Purity (%) Expiration Date NA 01/01/10 The molecular structure of the reference material is given below. PFOS Chemical Name Molecular weight = = Perfluorooctanesulfonate 499 (CgFnSOj-) O C8F17S -- o o Note: The neutral molecule and standard form from which PFOS (anion) is derived, is potassium perfluorooctanesulfonate [CgFnSC^K], molecular weight 538. 6.0 DESCRIPTION OF ANALYTICAL METHOD 6.1 Extraction Procedure Each sample was diluted in methanol prior to analysis by LC/MS/MS electrospray. 6.2 Preparation of Standards and Fortification Solutions Standard solutions of PFOS were prepared on January 10, 2002. An individual stock standard solution was prepared at a concentration of 100 pg/mL by dissolving 10 mg of the standard (corrected for purity and/or salt content) in methanol. From this solution, a 1.0 pg/mL fortification standard solution was prepared by taking 0.1 mL of the stock and bringing the volume up to 10 mL with methanol. A 0.1 pg/mL fortification standard was prepared by taking 1.0 mL of the 1.0 pg/mL fortification standard and bringing the volume up to 10 mL with methanol. A 0.01 pg/mL standard was prepared by taking 0.1 mL of the 0.1 pg/mL standard and bringing to 10 mL with methanol. Exygen Research 000479 Page 10 of 21 418-018:PAGE F -l 1 Exygen Study No.: 023-069 A set of standards containing PFOS was prepared by dilution of the 0.1 pg/mL various calibration solutions in the following manner: Initial Cone. (pg/mL)1 0.1 0.1 0.1 0.005 0.002 0.001 0.001 Volume (mL) 0.5 0.2 0.1 1.0 1.0 1.0 0.5 Diluted to (mL) 10 10 10 10 10 10 10 Final Cone. (pg/mL) 0.005 0.002 0.001 0.0005 0.0002 0.0001 0.00005 The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4 2C) when not in use. Documentation of standard preparation can be found in the raw data associated with this report. 6.3 Chromatography Quantification of PFOS was accomplished by LC/MS/MS electrospray. The retention time of PFOS was ~ 4.4 min. 6.4 Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 0.0001 pg/mL of PFOS. 6.5 Description of Instrument and Operating Conditions Instrument: Micromass Quattro Ultima Computer: Dell UltraScan PI 110 Software: COMPAQ Professional Workstation AP200 Windows NT, version 4 HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven Exygen Research 000480 Page 11 of 21 418-018:PAGE F-12 Exygen Study No.: 023-069 HPLC Column:Genesis C8(Jones Chromatography), 2.1 mm x 50 mm, 4p Column Temp.: 35 C Injection Voi.: 10 pL Mobile Phase (A): 2 mM Ammonium Acetate in ASTM type I water Mobile Phase (B): Methanol Flow Rate: 0.3 mL/min. Time % A % B 0 60 40 1.0 0 100 7.0 0 100 7.5 60 40 11.0 60 40 Ions monitored: Analvte PFOS Mode negative Transition Monitored 499 ^ 99 Approximate Retention Time (min) 4.4 Tune File Parameters Controls IS-Iospray DP-Declustering Potential FP-Focusing Potential EP-Entrance Potential CE-Collision Energy CXP-Collision Cell Exit Potential DF-Deflector CEM-Channel Electron Multiplier Set -4200.0 -51.0 -230.0 10.0 -70 -5 300.0 2800.0 Gas Flows Nebulizer Gas Curtain Gas Collision Gas TIS Temperature Set 12 13 4 350C 6.6 Quantitation and Example Calculation Twenty microliters of sample or calibration standard were injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x fit weighted linear regression) by the Micromass software using six concentrations of standards. The concentration was determined from the equations below. Exygen Research Page 12 of 21 000481 418-018:PAGE F-13 Exygen Study No.: 023-069 Equation 1 calculated the amount of analyte found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Micromass software program. Then Equation 2 calculated the amount of analyte found in pg/mL (the equations for serum are shown). Equation 1: Equation 2: Analyte found (ng/mL) = (Peak area - intercept) slope Analyte found (pg/mL) = (anal, found (ng/mL) x DF) x 1 ug 1000 ng Where: DF = Dilution Factor Equation 3 calculated the percent recovery. Equation 3: Recovery (%) = /anal, found ('ug/mLl x 100 sample cone. (pg/mL) An example of a calculation using an actual sample follows: Dosing solution sample Exygen ID 0200283 (Set 021402A), where: peak area intercept slope dilution factor sample cone. = 1380 = 41.3545 = 721.773 = 200,000 = 400 pg/mL From equation 1: Analyte found (ng/mL) = 11380 - 41.35451 721.773 = 1.85 ng/mL Exygen Research Page 13 of 21 000482 418-018:P A G E F -14 Exygen Study No.: 023-069 From equation 2: Analyte found (pg/mL) = (1.85 ng/mL x 200,000) x 1 |ig 1000 ng = 370 |ig/mL From equation 3: % Recovery = 370 pg/mL x 100 400 pg/mL = 93% Note: This example calculation was done using rounded numbers, and therefore may be slightly different from the values shown in the RAW DATA. 7.0 EXPERIMENTAL DESIGN The set of samples consisted of an instrument blank, a set of calibrations at the beginning and throughout the run and the 27 samples. 8.0 RESULTS Individual recoveries for PFOS in the dosing solution samples are detailed in Table I. The average percent recovery standard deviation for PFOS in the dosing solutions was 93% 7%. A typical calibration curve and representative chromatograms of a standard and sample are given in Figures 1-3. 9.0 CONCLUSIONS The concentration and homogeneity of the dosing solutions were demonstrated by the results of this report. 10.0 RETENTION OF DATA AND SAMPLES When the final report is complete, all original paper data generated by Exygen Research will be shipped to the sponsor. This does not include facility-specific raw data such as instrument logs. Exact copies of all raw data, as well as a signed copy of the final Exygen Research Page 14 of 21 000483 418-018:PAGE F-15 Exygen Study No.: 023-069 analytical report and all original facility-specific raw data, will be retained in the Exygen Research archives for the period of time specified in 21 CFR Part 58. Retained samples of reference substances are archived by the sponsor. Exygen Research Page 15 of 21 000484 418-018:PAGE F-16 Exygen Study No.: 023-069 TABLES Exygen Research m485 Page 16 of 21 418-018:PAGE F-17 Exygen Study No.: 023-069 Table I. Summary of PFOS in Dosing Solution Samples Exygen ID 0200283 0200284 0200285 0200286 0200287 0200288 0200289 0200290 0200291 0200292 0200293 0200294 0200295 0200296 0200297 0200298 0200299 0200300 0200301 0200302 0200303 0200304 0200305 0200306 0200307 0200308 0200309 Sponsor ID 1 o f 6T (0.4 mg/mL) 28.AUG.00 3 o f 6M (0.4 mg/mL) 28.AUG.00 5 o f 6B (0.4 mg/mL) 28.AUG.00 1 o f 2 (0 mg/mL) I8.SEP.00 1 of 2 (0.08 mg/mL) 18.SEP.00 I o f 2 (0.16 mg/mL) 18.SEP.00 1 o f 2 (0.20 mg/mL) 18.SEP.00 1 of 2 (0.24 mg/mL) 18.SEP.00 1 of 2 (0.32 mg/mL) 18.SEP.00 1 of 2 (0.40 mg/mL) 18.SEP.00 1 o f 6T (0.4 mg/mL) 27.SEP.00 3 of 6M (0.4 mg/mL) 27.SEP.00 5 of 6B (0.4 mg/mL) 27.SEP.00 1 o f 2 ( 0 mg/mL) 01.NOV.OO 1 o f 2 (0.08 mg/mL) 01.NOV.OO 1 o f 2 (0.16 mg/mL) 01.NOV.OO 1 o f 2 (0.20 mg/mL) 01.NOV.OO 1 o f 2 (0.24 mg/mL) 01.NOV.OO 1 o f 2 (0.32 mg/mL) 01.NOV.OO 1 o f 2 (0.40 mg/mL) 01 .NOV.OO 1 o f 2 (0 mg/mL) 15.NOV.OO 1 o f 2 (0.08 mg/mL) 15.NOV.OO 1 o f 2 (0.16 mg/mL) 15.NOV.OO 1 o f 2 (0.20 mg/mL) 15.NOV.OO 1 o f 2 (0.24 mg/mL) 15.NOV.OO 1 o f 2 (0.32 mg/mL) 15.NOV.OO 1 o f 2 (0.40 mg/mL) 15.NOV.00 DF 200000 200000 200000 20 50000 50000 200000 200000 200000 200000 200000 200000 200000 20 50000 50000 200000 200000 200000 200000 20 50000 50000 200000 200000 200000 200000 Peak Area 1380 1522 1551 ND 995 2156 714 867 1079 1385 1495 1423 1313 91 1002 2046 743 863 1054 1332 98 1001 2082 802 826 1145 1481 Analyte Found (ng/mL) 1.85 2.05 2.09 ND 1.32 2.93 0.932 1.14 1.44 1.86 2.01 1.91 1.76 NQ 1.33 2.78 0.972 1.14 1.40 1.79 NQ 1.33 2.83 1.05 1.09 1.53 1.99 Analyte Sample Found Cone. (pg/mL) (pg/mL) 371 400 410 400 418 400 ND 66.1 ' 80 146 160 186 200 229 240 288 320 372 400 403 400 383 400 352 400 NQ 66.5 80 139 160 194 200 228 240 281 320 358 400 NQ 66.5 80 141 160 211 200 217 240 306 320 399 400 Rec. (%) 93 103 105 - 83 92 93 95 90 93 101 96 88 - 83 87 97 95 88 89 - 83 88 105 91 96 100 ND = Not Detected NQ = Not Quantifiable (A peak was detected at the corresponding analyte retention time but was less than the lowest concentration of the calibration standards (0.1 ng/mL)) Exygen Research 000486 Page 17 of 21 418-018:PAGE F-18 Exygen Study No.: 023-069 FIGURES Exygen Research 000487 Page 18 of 21 418-018:PAGE F-19 Exygen Study No.: 023-069 Figure 1. Typical Calibration Curve for PFOS Compound 1 name: PFOS Coefficient of Determination: 0.999102 Calibration curve: 721.773 * x + 41.3545 Response type: External Std, Area Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None Exygen Research 000488 Page 19 of 21 418-018:PAGE F-20 Exygen Study No.: 023-069 Figure 2. Chromatogram Representing a 0.5 ng/mL Calibration Standard for PFOS C021402-4, 0.5 ng/mL Standard 021402A-104 Sm (Mn, 3x2) 15-Feb-2002 14:35:32 LC/MS/MS #7 MRM of 1 Channel ES- Exygen Research Page 20 of 21 000489 418-018:PAGE F-21 Exygen Study No.: 023-069 Figure 3. Chromatogram Representing Dosing Solution Sample (Exygen ID: 0200283, Set: 021402A) 0200283, DF = 200,000 021402A-109 Sm (Mn, 3x2) 15-Feb-2002 15:39:02 LC/MS/MS #7 MRM of 1 Channel ES- Exygen Research 00040 Page 21 of 21 APPENDIX G TEMEPRATURE AND RELATIVE HUMIDITY REPORTS 000491 ARGUS 418-018-.PAGE G -l Temperature and Relative Humidity Report Location: Room 17 Protocol Number: 418018 Range of Dates: 22-Aug-2000 14:44 to 12-Nov-2000 08:59 Target Range: Species: rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64F to 79F 83 1962.0 1956 Relative Humidity 30% to 70% 83 1962.0 1956 Mean ( SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 70.6 73.9 71.0 63.8 1954 0 2 (1.3) (99.9) (0.0) (0.1) 56.6 70.6 57.0 40.7 1955 1 0 ( 4.8) (99.9) (0.1) (0.0) Report Generated: 30-Nov-2000 at 14:02 COMMENTS: REVIEWED BY: DATE: 3 ' 2 & o / 000492 ARGUS 418-018:PAGE G-2 Temperature Deviations Report Location: Room 17 Protocol Number: 418018 Range of Dates: 22-Aug-2000 14:44 to 12-Nov-2000 08:59 Temperature Target Range: Species: rat Date 0 7 -S e p -2 0 0 0 0 7 -S e p -2 0 0 0 Time 06:00 07:00 Temp. 6 3 .8 L 6 3 .8 L 64F to 79F Date Time Temp. H = Value out of range - High L = Value out of range - Low Temp. = Temperature F Report Generated: 30-Nov-2000 at 14:10 These deviations did not adversely affect the outcome or interpretation of the study. The following deviation(s) impacted on the outcome of the study as described: 000493 ARGUS 418-018:PAGE G-3 Relative Humidity Deviations Report Location: Room 17 Protocol Number: 418018 Range of Dates: 22-Aug-2000 14:44 to 12-Nov-2000 08:59 Humidity Target Range: Species: rat Date 04-O ct-2000 Time 22:00 R.H. 70.6 H 30% to 70% Date Time H = V alu e o u t o f ran g e - H igh L = Value out o f range - Low R.H. = R elative H um idity (% ) R ep o rt G enerated: 30-N ov-2000 at 14:18 These deviations did not adversely affect the outcome or interpretation of the study. The following deviation(s) impacted on the outcome of the study as described: 000494 ARGUS 418-018:PAGE G-4 Temperature and Relative Humidity Report Location: Room 18 Protocol Number 418-018 Range of Dates: 05-Sep-2000 14:00 to 03-Dec-2000 13:59 Target Range: Species: rat Total Number of Days: Total Number of Hours: Total Number o f Data Points: Temperature 64F to 79F 90 2135.75 2129 Relative Humidity 30% to 70% 90 2135.75 2129 Mean ( SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 69.2 70.8 69.2 67.6 2129 0 0 ( 0.5) (100.0) (0.0) (0.0) 53.6 70.2 53.3 42.3 2128 1 0 (4.1) (100.0) (0.0) (0.0) Report Generated: 27-Mar-2001 at 18:18 COMMENTS: REVIEWED BY: DATE: ?> 000495 ARGUS 418-018-.PAGE G -5 Relative Humidity Deviations Report Location: Room 18 Protocol Number: 418-018 Range of Dates: 05-Sep-2000 08:00 to 03-Dec-2000 13:59 Humidity Target Range: Species: rat Date Time 05-0ct-2000 01:00 R.H. 70.2 H 30% to 70% Date Time R.H. H = Value out o f range - High L = Value out o f range - Low R.H. = Relative H um idity (% ) R eport Generated: 13-Dec-2001 at 14:57 These deviations did not adversely affect the outcome or interpretation of the study. The following deviation(s) impacted on the outcome of the study as described: 000496 APPENDIX H BIOANALYTICAL REPORTS (ANILYTICS) 000497 418-018:PAGE H-l Client omcs INCORPORATED 200 Girard Street, Suite 200. Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: .01/16/2001 Client No., 1028 Study: 418018 DAMS GD21 Species: RAT l i i 1o ii *o Accession Spec. No. ID B 0030145 10901 B 0030146 10902 B 0030147 10903 B 0030148 10904 B 0030149 10905 B 0030150 10907 B 0030151 10908 B 0030152 10910 Mean S.D. Group 1 Sex Age CHOLEST GLUCOSE LDL-DIR HDL-DIR TRIG MG/DL MG/DL MG/DL MG/DL MG/DL 1F 1F 1F 1F 1F 1F 1F 1F 95 117 12 44 322 79 116 10 61 151 82 89 7 65 116 77 87 7 51 176 79 99 15 54 156 97 90 1 13 771 103 94 2 18 559 62 99 3 40 257 84.3 13.3 98.9 11.7 7.1 5 43.3 19 313.5 233.6 B 0030201 11040 B 0030202 11044 B 0030203 11045 B 0030204 11046 B 0030205 11047 B 0030206 11050 B 0030207 11051 B 0030208 11052 Mean S.D. Group 12 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 88 76 84 77 75 84 55 86 78.1 10.5 103 93 111 98 119 106 104 105 104.9 7.8 3 12 14 17 13 9 7 11 10.8 4.4 58 42 50 27 53 51 33 34 43.5 11.2 240 132 171 313 127 174 153 244 194.3 65.2 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 104 Page 1 + 000498 418-018:PAGE H-2 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT TJ 11 1 O1 11 11 Accession Spec. No. ID B 0030209 11053 B 0030210 11054 B 0030211 11056 B 0030212 11058 B 0030213 11060 B 0030214 11062 B 0030215 11063 B 0030216 11066 Mean S.D. Group 13 1 1 Sex Age 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F CHOLEST MG/DL 64 73 83 69 70 66 75 78 72.3 6.3 GLUCOSE MG/DL 78 76 79 129 103 98 103 104 96.3 18 LDL-DIR MG/DL 9 7 15 8 9 15 11 16 11.3 3.6 HDL-DIR MG/DL 43 26 44 34 59 26 45 51 41 11.7 TRIG MG/DL 158 318 152 270 79 296 216 119 201 87.5 B 0030153 10915 B 0030154 10916 B 0030155 10917 B 0030156 10918 B 0030157 10919 B 0030158 10920 B 0030159 10925 B 0030160 10928 Mean S.D. Group 2 2F 2F 2F 2F 2F 2F 2F 2F 79 76 90 88 99 83 119 106 92.5 14.6 121 108 110 111 112 121 101 84 108.5 11.9 4 6 2 10 0 1 0 0 2.9 3.6 46 62 19 55 16 35 17 9 32.4 20 249 85 459 203 719 337 552 1027 453.9 307.8 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 104 Page 2 + 0004S3 418-018-.PAGE H-3 \L A j L^LL JL JL V-/K_y 200 Girard Street, Suite 200, Gaithersburg, MD 20877 INCORPORATED 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No,. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Sgec. No. B 0030161 10930 B 0030162 10931 B 0030163 10932 B 0030164 10933 B 0030165 10934 B 0030166 10935 B 0030167 10937 B 0030168 10940 Mean S.D. Group 3 Gp Sex Age 3F 3F 3F 3F 3F 3F 3F 3F CHOLEST MG/DL 76 75 131 85 90 117 74 80 91 21.4 GLUCOSE MG/DL 118 102 113 107 93 137 87 108 108.1 15.4 LDL-DIR MG/DL 10 11 31 15 1 14 6 6 11.8 9 HDL-DIR MG/DL 49 47 26 33 23 49 35 19 35.1 12.1 TRIG MG/DL 228 221 439 226 407 279 294 353 305.9 85.1 B 0030169 10945 B 0030170 10946 B 0030171 10947 B 0030172 10949 B 0030173 10950 B 0030174 10951 B 0030175 10953 B 0030176 10954 Mean S.D. Group 4 4F 4F 4F 4F 4F 4F 4F 4F 134 74 94 53 84 87 100 85 88.9 23.1 86 88 125 110 108 102 107 122 106 14 0 9 10 6 17 13 12 10 9.6 5 11 56 48 50 56 61 59 41 47.8 16.2 887 115 197 42 113 93 259 243 243.6 270.8 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 104 Page 3 + 000500 418-018:PAGE H-4 O INCORPORATED 200 Girard Street, Suite 200, Gaithersburg. MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Spec. No. ID B 0030177 10958 B 0030178 10959 B 0030179 10960 B 0030180 10961 B 0030181 10962 B 0030182 10963 B 0030183 10964 B 0030184 10965 Mean S.D. Group 5 Gp Sex Age 5F 5F 5F 5F 5F 5F 5F 5F CHOLEST MG/DL 113 114 72 68 75 75 108 92 89.6 19.6 GLUCOSE MG/DL 115 91 125 99 110 58 108 97 100.4 20.2 LDL-DIR MG/DL 2 9 8 11 13 10 15 0 8.5 5.2 HDL-DIR MG/DL 16 16 31 33 47 36 30 15 28 11.5 TRIG MG/DL 531 583 133 213 140 220 280 690 348.8 218.6 B 0030185 10972 B 0030186 10974 B 0030187 10975 B 0030188 10976 B 0030189 10978 B 0030190 10980 B 0030191 10981 B 0030192 10982 Mean S.D. Group 6 6F 6F 6F 6F 6F 6F 6F 6F 82 78 77 71 76 72 77 101 79.3 9.4 90 79 140 97 103 85 96 98 98.5 18.5 14 13 4 11 9 5 9 14 9.9 3.9 33 25 54 43 44 27 37 65 41 13.6 122 265 161 143 175 344 214 111 191.9 79.3 Reference Range 48 - 74 - 0 0 3- 96 319 0 0 104 Page 4 + 0G0591 418-018:PAGE H-5 200 Girard Street. Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 INC. Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT ip> i 11O 11 11 11 Accession Spec. No. ID B 0030193 10985 B 0030194 10987 B 0030195 10988 B 0030196 10989 B 0030197 10990 B 0030198 10991 B 0030199 10992 B 0030200 10993 Mean S.D. Group 7 Sex Age CHOLEST GLUCOSE LDL-DIR HDL-DIR TRIG MG/DL MG/DL MG/DL MG/DL MG/DL 7F 7F 7F 7F 7F 7F 7F 7F 89 95 16 61 111 79 109 7 61 95 83 99 30 37 196 81 145 12 69 48 63 100 7 49 140 72 109 10 34 294 94 84 1 12 632 62 110 13 50 91 77.9 11.5 106.4 17.9 12 8.6 46.6 18.4 200.9 190.1 Reference Range 48 96 74 319 Page 5 + 00 0- 30 104 000502 418-018:PAGE H-6 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client ARGUS RESEARCH LABORATORIES, INC 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported:. 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Sgec. No. B 0030145 10901 B 0030146 10902 B 0030147 10903 B 0030148 10904 B 0030149 10905 B 0030150 10907 B 0030151 10908 B 0030152 10910 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL p g /m l 1.45 0.54 0.12 0.76 0.95 0.65 0.20 0.00 86.51 76.54 62.42 66.55 62.61 116.76 75.48 44.96 0.74 0.76 0.41 0.51 0.73 0.77 0.52 0.19 .584 .482 73.979 .579 21.234 .209 TSH NG/ML 3.22 2.09 1.63 1.32 1.87 2.14 1.84 0.41 1.815 .794 FREE T4 NG/DL 0.39 0.39 0.28 0.35 0.38 0.22 0.23 0.15 .299 .092 B 0030201 11040 B 0030202 11044 B 0030203 11045 B 0030204 11046 B 0030205 11047 B 0030206 11050 B 0030207 11051 B 0030208 11052 Mean S.D. Group 12 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 65.52 43.96 42.05 44.97 39.63 51.85 34.20 57.84 47.503 10.268 0.19 0.06 0.24 0.06 0.10 0.00 0.33 .14 .117 3.79 1.11 2.43 1.47 1.20 2.12 4.30 2.346 1.263 0.15 0.06 0.06 0.04 0.05 0.05 0.03 0.09 .066 .038 Reference Range 3 - 50 7 100 Page 6 + 00 .57 3.41 1.9 2.5 000503 418-018:PAGE H-7 fL/m Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Spec. No. ID B 0030209 11053 B 0030210 11054 B 0030211 11056 B 0030212 11058 B 0030213 11060 B 0030214 11062 B 0030215 11063 B 0030216 11066 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F T4 ,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 33.47 54.87 42.95 52.33 35.61 36.45 47.52 36.32 0.02 0.32 0.16 0.40 0.00 0.20 0.00 0 42.44 .157 0 8.271 .161 TSH NG/ML 1.52 2.07 1.37 3.21 1.22 1.21 0.00 1.514 .973 FREE T4 NG/DL 0.04 0.05 0.04 0.08 0.10 0.03 0.05 0.05 .055 .023 B 0030153 10915 B 0030154 10916 B 0030155 10917 B 0030156 10918 B 0030157 10919 B 0030158 10920 B 0030159 10925 B 0030160 10928 Mean S.D. Group 2 2F 2F 2F 2F 2F 2F 2F 2F 0.69 0.22 1.58 0.14 0.23 0.27 0.16 0.02 .414 .51 73.81 56.28 104.24 66.85 70.56 80.36 72.56 63.91 73.571 14.314 0.34 0.26 0.42 0.78 0.76 0.60 0.44 .514 .203 1.78 2.75 2.03 3.88 4.12 1.20 2.627 1.177 0.42 0.28 0.45 0.29 0.26 0.26 0.27 0.14 .296 .098 Reference Range 3 - 50 7 100 Page 7 + 0 0 .57 - 1.9 - 3.41 2.5 000294 418-018:PAGE H-8 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Sgec. No. B 0030161 10930 B 0030162 10931 B 0030163 10932 B 0030164 10933 B 0030165 10934 B 0030166 10935 B 0030167 10937 B 0030168 10940 Mean S.D. Group 3 Gp Sex Age 3F 3F 3F 3F 3F 3F 3F 3F T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 0.02 0.00 0.00 0.00 0.00 48.86 44.68 55.32 59.16 62.09 62.03 52.27 47.93 0.06 0.38 0.12 0.43 0.43 0.44 0.31 0.33 .003 .007 54.043 .313 6.67 .146 TSH NG/ML 1.96 2.40 2.12 2.05 1.58 1.91 1.20 1.889 .391 FREE T4 NG/DL 0.03 0.05 0.04 0.14 0.09 0.12 0.06 0.02 .069 .044 B 0030169 10945 B 0030170 10946 B 0030171 10947 B 0030172 10949 B 0030173 10950 B 0030174 10951 B 0030175 10953 B 0030176 10954 Mean S.D. Group 4 4F 4F 4F 4F 4F 4F 4F 4F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 65.48 41.50 46.55 38.85 36.70 50.05 45.31 46.82 46.408 8.895 0.55 0.07 0.25 0.12 0.00 0.15 0.09 0.24 .184 .17 4.56 2.05 1.79 1.78 0.94 3.16 2.42 2.30 2.375 1.086 0.04 0.03 0.08 0.10 0.07 0.14 0.06 0.06 .073 .035 Reference Range 3 - 50 7 100 Page 8 + 0 0 .57 - 1.9 3.41 2.5 000505 418-018:PAGE H-9 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 INC. Date Collected: Date Received: 12/08/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Sgec. No. B 0030177 10958 B 0030178 10959 B 0030179 10960 B 0030180 10961 B 0030181 10962 B 0030182 10963 B 0030183 10964 B 0030184 10965 Mean S.D. Group 5 Gp Sex Age 5F 5F 5F 5F 5F 5F 5F 5F T4,TOTAL Tl,TOTAL FREE T3 UG/DL NG/DL PG/ML 1.29 0.44 0.00 1.27 0.81 0.12 0.00 0.39 91.13 80.57 27.60 81.69 98.93 86.18 67.30 81.80 0.85 0.77 0.86 0.68 0.63 0.16 0.91 .54 .529 76.9 .694 21.899 .256 TSH NG/ML 3.72 2.54 2.64 2.14 1.68 2.544 .759 FREE T4 NG/DL 0.30 0.21 0.36 0.52 0.16 0.21 0.21 .281 .125 B 0030185 10972 B 0030186 10974 B 0030187 10975 B 0030188 10976 B 0030189 10978 B 0030190 10980 B 0030191 10981 B 0030192 10982 Mean S.D. Group 6 6F 6F 6F 6F 6F 6F 6F 6F 0.10 0.00 0.00 0.00 0.00 0.00 0.00 0.00 .013 .035 64.17 49.59 62.37 61.84 45.53 41.40 48.59 42.76 52.031 9.331 0.15 0.26 0.36 0.27 0.17 0.30 0.37 0.00 .235 .123 2.72 0.94 0.76 1.28 1.53 1.48 1.452 .691 0.21 0.04 0.27 0.17 0.13 0.02 0.16 0.07 .134 .086 Reference Range 3 - 50 7 100 Page 9 + 00 .57 3.41 1.9 2.5 00050 418-018:PAGE H-10 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Sgec. No. B 0030193 10985 B 0030194 10987 B 0030195 10988 B 0030196 10989 B 0030197 10990 B 0030198 10991 B 0030199 10992 B 0030200 10993 Mean S.D. Group 7 Gp Sex Age 7F 7F 7F 7F 7F 7F 7F 7F T4 ,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 51.27 51.82 32.73 59.37 43.11 44.67 56.54 52.65 0.08 0.16 0.13 0.29 0.27 0.08 0.62 0.34 0 49.02 .246 0 8.527 .18 TSH NG/ML 2.61 1.74 1.12 1.02 2.47 3.12 2.013 .854 FREE T4 NG/DL 0.10 0.14 0.04 0.29 0.09 0.04 0.05 0.13 .11 .082 Reference Range 3- 50 - 0- .57 - 1.9 - 7 100 0 3.41 Approved --Page 10 O f / ? ,'xZl____ 10 D a t e -- l L & J j IL 0007 418-018:PAGE H-ll Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 04/17/2001 Client No.. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0030547 10906 B 0030548 10909 B 0030549 10911 B 0030550 10913 B 0030551 10914 B 0030630 10912 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F CHOLEST MG/DL 63 84 82 57 80 81 74.5 11.5 GLUCOSE MG/DL 152 155 110 146 148 192 150.5 26.1 LDL-DIR MG/DL 0 2 0 4 0 1 1.2 1.6 HDL-DIR MG/DL 28 54 21 45 27 66 40.2 17.7 TRIG MG/DL 246 141 512 88 177 147 218.5 152.8 B 0030589 11019 B 0030590 11021 B 0030591 11020 B 0030592 11023 B 0030593 11024 B 0030594 11025 B 0030595 11026 B 0030596 11027 B 0030597 11028 Mean S.D. Group 10 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 72 56 74 41 51 58 56 49 52 56.6 10.6 159 187 154 183 170 230 181 153 165 175.8 23.9 5 5 1 3 4 4 5 6 0 3.7 2 57 44 32 33 46 49 46 39 36 42.4 8.2 96 67 259 98 45 70 60 60 178 103.7 70.2 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 104 Page 1 + O005&S 418-018:PAGE H -12 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/ 12/2000 Date Reported: 04/17/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0030598 11029 B 0030599 11031 B 0030600 11032 B 0030601 11033 B 0030602 11034 B 0030603 11035 B 0030604 11036 B 0030605 11037 B 0030606 11038 Mean S.D. Group 11 Gp Sex Age 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F CHOLEST MG/DL 50 58 61 53 44 44 50 58 69 54.1 8.2 GLUCOSE MG/DL 168 172 156 174 179 183 150 150 193 169.4 15 LDL-DIR MG/DL 1 4 2 4 0 0 0 0 3 1.6 1.7 HDL-DIR MG/DL 31 46 43 40 37 31 17 36 55 37.3 10.7 TRIG MG/DL 243 115 70 87 132 186 370 195 60 162 99.4 B 0030607 11041 B 0030608 11049 B 0030622 11042 B 0030623 11048 Mean S.D. Group 12 12 F 12 F 12 F 12 F 53 38 70 80 60.3 18.6 172 195 212 175 188.5 18.7 5 6 1 8 5 2.9 42 37 41 61 45.3 10.7 58 39 234 87 104.5 88.6 B 0030609 11059 Reference Range 13 F 49 48 96 158 74 319 2 00 42 00 104 3104 Page 2 + 000509 418-018:PAGE H-13 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax : 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 04/17/2001 Client No . 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0030624 11055 B 0030625 11057 B 0030626 11061 B 0030627 11064 B 0030628 11065 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F 13 F 13 F CHOLEST MG/DL 58 63 74 72 61 62.8 9.2 GLUCOSE MG/DL 188 156 277 193 157 188.2 46.5 LDL-DIR MG/DL 7 7 5 6 3 5 2.1 HDL-DIR MG/DL 48 46 70 64 52 53.7 11 TRIG MG/DL 48 67 89 72 90 78.3 20 B 0030552 10921 B 0030553 10922 B 0030554 10923 B 0030555 10924 B 0030556 10927 Mean S.D. Group 2 2F 2F 2F 2F 2F 68 49 78 70 105 74 20.3 132 171 134 144 151 146.4 15.8 0 0 1 11 2 2.8 4.7 38 30 36 47 65 43.2 13.6 188 202 131 52 151 144.8 59.1 B 0030557 10929 B 0030558 10938 B 0030559 10942 B 0030610 10941 Mean S.D. Group 3 3F 3F 3F 3F 53 51 53 44 50.3 4.3 141 149 143 161 148.5 9 2 2 0 3 1.8 1.3 45 37 40 34 39 4.7 82 96 165 90 108.3 38.3 Reference Range 48 96 74 319 Page 3 + 00 0 0 3104 00051 418-018:PAGE H-14 M 4 l-Jl JL 1 V A - / 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C O R P O R A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported:. 04/17/2001 Client No.. 1028 Study: 418018 DAMS LD5 Species : RAT Accession Sgec. No. B 0030611 10943 B 0030612 10948 B 0030613 10952 B 0030614 10955 B 0030615 10956 B 0030629 10944 Mean S.D. Group 4 Gp Sex Age 4F 4F 4F 4F 4F 4F CHOLEST MG/DL 69 56 77 66 76 65 68.2 7.8 GLUCOSE MG/DL 184 179 194 138 235 158 181.3 33.1 LDL-DIR MG/DL 3 0 4 3 6 13 4.8 4.4 HDL-DIR MG/DL 48 46 54 50 65 48 51.8 7 TRIG MG/DL 93 192 107 141 87 43 110.5 51 B 0030560 10957 B 0030561 10966 B 0030562 10967 B 0030563 10968 B 0030564 10969 B 0030565 10970 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 59 74 66 72 72 54 66.2 8.1 152 123 163 144 161 136 146.5 15.4 1 0 6 0 2 0 1.5 2.3 27 51 45 32 45 19 36.5 12.4 230 159 49 252 143 288 186.8 87.2 B 0030566 10977 B 0030567 10979 Reference Range 6F 6F 44 45 48 96 135 142 74 319 0 3 00 33 34 00 135 102 3104 Page 4 + 000511 418-018:PAGE H-15 ^>UJL-*LL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: . 04/17/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0030568 10983 B 0030616 10971 B 0030617 10973 B 0030618 10984 Mean S.D. Group 6 Gp Sex Age 6F 6F 6F 6F CHOLEST MG/DL 57 81 45 69 56.8 15.3 GLUCOSE MG/DL 152 176 163 209 162.8 26.9 LDL-DIR MG/DL 3 12 6 3 4.5 4.1 HDL-DIR MG/DL 45 53 38 57 43.3 10.1 TRIG MG/DL 62 96 42 84 86.8 32.5 B 0030569 10995 B 0030570 10997 B 0030619 10986 B 0030620 10994 B 0030621 10996 Mean S.D. Group 7 7F 7F 7F 7F 7F 41 40 77 47 68 54.6 16.9 192 196 176 158 147 173.8 21.2 4 3 9 6 3 5 2.5 33 35 59 41 54 44.4 11.6 78 75 72 50 116 78.2 23.8 B 0030571 10999 B 0030572 11000 B 0030573 11001 B 0030574 11002 B 0030575 11003 Reference Range 8F 8F 8F 8F 8F 77 57 51 56 69 48 96 127 164 216 146 153 74 319 1 6 2 3 1 00 52 46 35 32 32 00 240 64 119 147 286 3104 Page 5 + 000512 418-018:PAGE H-16 cA N I' INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: .04/17/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Spec. No. lu B 0030576 11004 B 0030577 11005 B 0030578 11006 B 0030579 11007 B 0030580 11008 Mean S.D. Group 8 Gp Sex Age 8F 8F 8F 8F 8F CHOLEST MG/DL 53 64 70 56 61 61.4 8.4 GLUCOSE MG/DL 168 150 185 133 147 158.9 26.2 LDL-DIR MG/DL 2 0 1 0 2 1.8 1.8 HDL-DIR MG/DL 45 32 38 37 38 38.7 6.8 TRIG MG/DL 79 260 154 243 153 174.5 78 B 0030581 11009 B 0030582 11010 B 0030583 11011 B 0030584 11012 B 0030585 11013 B 0030586 11015 B 0030587 11016 B 0030588 11018 Mean S.D. Group 9 9F 9F 9F 9F 9F 9F 9F 9F 50 56 51 74 58 47 45 63 55.5 9.5 158 135 144 144 148 179 150 183 155.1 17.3 4 0 1 6 0 1 4 4 2.5 2.3 32 43 33 42 24 35 35 52 37 8.5 116 167 164 100 324 195 89 85 155 79.5 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 104 Page 6 + 000513 418-018:PAGE H -17 ^ -i i tJ l -X INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 04/17/2001 Client No. 1028 Study: 418018 DAMS LD5 Species : RAT ia i !O 1 11 1! 11 Accession Spec. No. l5 B 0030547 10906 B 0030548 10909 B 0030549 10911 B 0030550 10913 B 0030551 10914 B 0030630 10912 Mean S.D. Group 1 Sex Age T4,TOTAL T3,TOTAL FREE T3 TSH UG/DL NG/DL PG/ML NG/ML 1F 1F 1F 1F 1F 1F 1.66 2.25 2.63 1.71 0.87 2.44 100.60 99.04 102.40 80.08 94.35 91.84 1.14 0.97 0.75 0.90 0.72 0.97 1.18 3.92 1.26 0.00 1.66 1.927 .648 94.718 .908 8.185 .156 1.604 1.435 FREE T4 NG/DL 0.88 0.84 1.08 0.57 0.65 0.82 .807 .18 B 0030589 11019 B 0030590 11021 B 0030591 11020 B 0030592 11023 B 0030593 11024 B 0030594 11025 B 0030595 11026 B 0030596 11027 B 0030597 11028 Mean S.D. Group 10 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 0.88 1.15 1.02 0.71 0.57 0.45 0.60 0.70 0.47 .728 .243 79.66 77.77 77.15 50.76 59.72 65.05 45.51 49.83 55.68 62.348 13.181 0.86 0.85 0.44 0.27 0.71 0.31 0.35 0.49 0.69 .552 .23 1.22 0.92 2.22 0.96 0.80 0.94 0.46 1.36 1.11 .523 0.31 0.47 0.32 0.21 0.25 0.35 0.19 0.19 0.20 .277 .095 Reference Range 3 - 50 7 100 Page 7 + 0 0 .57 - 1.9 - 3.41 2.5 000514 418-018:PAGE H-18 ^4i L , J I JL J L INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 04/17/2001 Client No . 1028 Study: 418018 DAMS LD5 Species: RAT Accession Spec. No. ID B 0030598 11029 B 0030599 11031 B 0030600 11032 B 0030601 11033 B 0030602 11034 B 0030603 11035 B 0030604 11036 B 0030605 11037 B 0030606 11038 Mean S.D. Group 11 Gp Sex Age 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.87 0.00 0.52 0.42 0.89 0.35 0.00 0.26 0.90 61.61 44.85 73.96 58.43 73.76 53.37 57.45 61.84 85.82 0.62 0.00 0.86 0.61 0.87 0.55 0.52 0.45 0.69 .468 .358 63.454 .574 12.409 .259 TSH NG/ML 1.46 1.56 0.60 1.76 2.53 0.50 0.82 0.00 1.154 .819 FREE T4 NG/DL 0.36 0.08 0.26 0.18 0.67 0.23 0.23 0.18 0.33 .28 .168 B 0030607 11041 B 0030608 11049 B 0030622 11042 B 0030623 11048 Mean S.D. Group 12 12 F 12 F 12 F 12 F 0.14 0.60 0.46 0.27 .368 .203 67.01 53.37 100.62 73.23 73.558 19.857 0.49 0.33 0.72 0.43 .493 .165 1.49 1.96 1.16 1.31 1.48 .347 0.22 0.29 0.32 0.26 .273 .043 B 0030609 11059 Reference Range 13 F 0.24 37 48.45 50 100 0.16 00 0.97 .57 3.41 0.15 1.9 2.5 Page 8 + 000515 418-018-.PAGEH-19 i * mJi -X. x w V w * 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C OR P O R A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 04/17/2001 Client No.. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0030624 11055 B 0030625 11057 B 0030626 11061 B 0030627 11064 B 0030628 11065 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F 13 F 13 F T4 ,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.23 0.00 0.33 0.26 0.60 61.96 72.68 76.74 66.25 83.98 0.76 0.64 0.50 0.41 .277 .194 68.343 .494 12.447 .23 TSH NG/ML 3.48 0.82 1.79 2.14 1.84 1.07 FREE T4 NG/DL 0.25 0.17 0.26 0.16 .198 .053 B 0030552 10921 B 0030553 10922 B 0030554 10923 B 0030555 10924 B 0030556 10927 Mean S.D. Group 2 2F 2F 2F 2F 2F 4.22 2.31 2.89 1.24 2.38 2.608 1.083 81.28 88.13 95.06 63.21 116.64 88.864 19.533 0.83 0.72 0.79 0.48 1.29 .822 .295 1.04 4.41 0.96 0.48 2.92 1.962 1.655 1.21 0.74 1.02 0.57 0.93 .894 .248 B 0030557 10929 B 0030558 10938 B 0030559 10942 B 0030610 10941 Mean S.D. Group 3 3F 3F 3F 3F 0.08 0.00 0.11 0.54 .183 .243 68.57 56.59 70.90 66.95 65.753 6.32 0.65 0.53 0.81 0.79 .695 .131 1.12 0.43 2.78 0.58 1.228 1.077 0.30 0.17 0.30 0.31 .27 .067 Reference Range 3 - 50 7 100 Page 9 + 00 .57 3.41 1.9 2.5 000516 418-018-.PAGEH-20 Client oxncs 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C OR P OR A T E D 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: . 04/17/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0030611 10943 B 0030612 10948 B 0030613 10952 B 0030614 10955 B 0030615 10956 B 0030629 10944 Mean S.D. Group 4 Gp Sex Age 4F 4F 4F 4F 4F 4F T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.24 0.32 0.77 0.23 0.39 0.04 67.82 74.34 94.97 81.22 73.68 64.92 0.82 0.75 0.58 0.74 0.30 .332 .245 76.158 .638 10.815 .208 TSH NG/ML 0.84 1.42 2.02 2.15 0.40 1.366 .751 FREE T4 NG/DL 0.35 0.21 0.16 0.22 0.17 .222 .076 B 0030560 10957 B 0030561 10966 B 0030562 10967 B 0030563 10968 B 0030564 10969 B 0030565 10970 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 2.80 3.23 1.51 1.69 1.35 1.94 2.087 .758 108.48 87.83 94.97 104.99 84.75 69.56 91.763 14.299 1.00 0.79 1.04 1.02 0.69 0.78 .887 .151 0.52 1.50 1.15 0.55 0.44 0.70 .81 .422 0.87 1.51 0.78 0.70 0.89 0.81 .927 .294 B 0030566 10977 B 0030567 10979 Reference Range 6F 6F 0.00 0.00 37 50.71 55.20 50 100 0.36 0.48 00 0.62 1.16 .57 3.41 0.21 0.27 1.9 2.5 Page 10 + 000513? 418-018:PAGE H-21 4, L J l A. JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM. PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 04/17/2001 Client No., 1028 Study: 418018 DAMS LD5 Species : RAT Accession Sgec. No. B 0030568 10983 B 0030616 10971 B 0030617 10973 B 0030618 10984 Mean S.D. Group 6 Gp Sex Age 6F 6F 6F 6F T4 ,TOTAL T 3 ,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.53 0.45 0.65 75.66 65.47 64.98 79.39 0.39 0.60 0.72 .272 .304 65.235 .51 11.143 .15 TSH NG/ML 0.68 2.02 1.92 1.28 .665 FREE T4 NG/DL 0.25 0.30 0.32 0.28 .272 .039 B 0Q30569 1Q995 B 0030570 10997 B 0030619 10986 B 0030620 10994 B 0030621 10996 Mean S.D. Group 7 7F 7F 7F 7F 7F 0.00 0.00 0.43 0.24 0.00 .134 .195 54.80 40.79 78.34 67.17 87.47 65.714 18.542 0.33 0.25 0.94 0.85 0.64 .602 .306 1.32 0.74 3.12 3.12 1.12 1.884 1.147 0.19 0.14 0.31 0.30 0.27 .242 .074 B 0030571 10999 B 0030572 11000 B 0030573 11001 B 0030574 11002 B 0030575 11003 Reference Range 8F 8F 8F 8F 8F 1.05 1.14 0.61 0.65 0.43 37 81.63 76.76 51.06 64.38 80.94 50 100 0.79 0.78 0.40 0.86 1.15 00 1.54 1.00 1.35 0.92 .57 3.41 0.47 0.50 0.41 0.64 0.30 1.9 2.5 Page 11 + 0G051* 418-018:PAGE H-22 l a -Ji JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 04/17/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0030576 11004 B 0030577 11005 B 0030578 11006 B 0030579 11007 B 0030580 11008 Mean S.D. Group 8 Gp Sex Age 8F 8F 8F 8F 8F T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 1.48 1.12 1.03 0.38 0.19 88.66 81.56 86.75 56.62 60.76 0.84 0.56 0.78 0.39 0.43 .808 .414 72.912 .698 13.47 .247 TSH NG/ML 1.36 1.02 1.06 1.15 0.88 1.142 .226 FREE T4 NG/DL 0.60 0.55 0.44 0.40 0.31 .462 .114 B 0030581 11009 B 0030582 11010 B 0030583 11011 B 0030584 11012 B 0030585 11013 B 0030586 11015 B 0030587 11016 B 0030588 11018 Mean S.D. Group 9 9F 9F 9F 9F 9F 9F 9F 9F 0.00 0.34 1.21 0.14 0.71 1.00 0.98 0.39 .596 .443 57.68 60.23 59.71 55.40 73.47 71.68 68.23 63.84 63.78 6.684 0.44 0.55 0.70 0.40 0.85 0.70 0.49 0.57 .588 .152 2.23 1.30 0.04 0.63 2.18 1.16 2.52 1.437 .918 0.22 0.28 0.26 0.21 0.26 0.34 0.39 0.27 .279 .06 Reference Range 3- 50 - 0- 7 100 . 0 Appro v e d ------ --2**'^!-------Page 12 of 12 .57 3.41 Date 1.9 2.5 -- 00051S 418-018:PAGE H-23 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/14/2000 Date Reported: . 01/16/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Sgec. No. B 0031037 10906 B 0031038 10909 B 0031039 10911 B 0031040 10913 B 0031041 10914 Mean S.D. Group 1 Gp Sex Age 1. F 1F 1F 1F 1F CHOLEST MG/DL 26 10 27 10 11 16.8 8.9 B 0031079 11019 B 0031080 11021 B 0031081 11020 B 0031082 11023 B 0031083 11024 B 0031084 11025 B 0031085 11026 B 0031086 11027 B 0031087 11028 Mean S.D. Group 10 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 17 0 2 38 0 0 41 17 23 15.3 16.3 Reference Range 4A8 Page 1 + 000529 418-018:PAGE H-24 cA N I' Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/14/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Sjsec. No. B 0031088 11029 B 0031089 11031 B 0031090 11032 B 0031091 11033 B 0031092 11034 B 0031093 11035 B 0031094 11036 B 0031095 11037 B 0031096 11038 Mean S.D. Group 11 Gp Sex Age 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F CHOLEST MG/DL 18 0 0 10 15 19 14 0 3 8.8 8.1 B 0031097 11041 B 0031098 11049 Mean S.D. Group 12 12 F 12 F 34 0 17 24 B 0031099 11059 Mean S.D. Group 13 13 F 0 0 0 Reference Range 48 96 Page 2 + 000521 418-018:PAGE H-25 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/14/2000 Date Reported: 01/16/2001 Client No.. 1028 Study: 418018 LD5 MILK Species: RAT Accession Spec. No. ID B 0031042 10921 B 0031043 10922 B 0031044 10923 B 0031045 10924 B 0031046 10927 Mean S.D. Group 2 Gp Sex Age 2F 2F 2F 2F 2F CHOLEST MG/DL 0 4 5 31 0 8 13.1 B 0031047 10929 B 0031048 10938 B 0031049 10942 Mean S.D. Group 3 3F 3F 3F 9 10 0 6.3 5.5 B 0031050 10957 B 0031051 10966 B 0031052 10967 B 0031053 10968 B 0031054 10969 B 0031055 10970 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 7 0 9 20 10 23 11.5 8.5 Reference Range 48 96 Page 3 + 000522 418-018:PAGE H-26 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/14/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Spec. No. ID B 0031056 10977 B 0031057 10979 B 0031058 10983 Mean S.D. Group 6 B 0031059 10995 B 0031060 10997 Mean S.D. Group 7 Gp Sex Age 6F 6F 6F CHOLEST MG/DL 0 6 7 4.3 3.8 7F 7F 47 61 54 9.9 B 0031061 10999 B 0031062 11000 B 0031063 11001 B 0031064 11002 B 0031065 11003 B 0031066 11004 B 0031067 11005 B 0031068 11006 B 0031069 11007 Reference Range 8F 8F 8F 8F 8F 8F 8F 8F 8F 12 18 41 18 22 25 9 19 3 48 96 Page 4 + 000523 418-018:PAGE H-27 Client c^/Wi omcs 200 Girard Street, Suite 200. Gaithersburg, MD 20877 I N C OR P OR A T E D 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/14/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Sgec. No. B 0031070 11008 Mean S.D. Group 8 Gp Sex Age 8F CHOLEST MG/DL 17 18.4 10.2 B 0031071 11009 B 0031072 11010 B 0031073 11011 B 0031074 11012 B 0031075 11013 B 0031076 11015 B 0031077 11016 B 0031078 11018 Mean S.D. Group 9 9F 9F 9F 9F 9F 9F 9F 9F 110 22 4 2 12 19 15 1 23.1 36 Reference Range 4O8A - Approved -- ____ ^ i l _ Page 5 of 5 D a t e --- L^Ll2./ 000524 418-018:PAGE H-28 Client mcs ^ / U \ U \ 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NC OR P OR A T E D 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE. BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Sgec. No. B 0030217 10901 B 0030218 10902 B 0030219 10903 B 0030220 10904 B 0030221 10905 B 0030222 10907 B 0030223 10908 B 0030224 10910 Mean S.D. Group 1 Gp Sex Age 1 1 1 1 1 1 1 1 CHOLEST MG/DL 40 55 58 60 49 43 58 43 50.8 8 GLUCOSE MG/DL 69 55 70 56 69 61 64 66 63.8 5.9 LDL-DIR MG/DL 15 34 35 35 25 17 32 25 27.3 8 HDL-DIR MG/DL 12 14 15 13 12 11 16 11 13 1.9 TRIG MG/DL 28 31 34 41 39 39 36 27 34.4 5.3 B 0030273 11040 B 0030274 11044 B 0030275 11045 B 0030276 11046 B 0030277 11047 B 0030278 11050 B 0030279 11051 B 0030280 11052 Mean S.D. Group 12 12 12 12 12 12 12 12 12 57 51 65 60 73 63 50 71 61.3 8.5 61 74 82 78 59 79 96 80 76.1 11.8 41 38 44 37 52 51 39 57 44.9 7.5 16 ' 12 14 13 15 16 14 16 14.5 1.5 19 26 38 36 38 21 25 20 27.9 8.2 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 125 Page 1 + 000525 418-01B-.PAGEH-29 MbJJLJLJL jL 200 Girard Street. Suite 200. Gaithersburg, MD 20877 I NC OR P OR A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Sgec. No. B 0030281 11053 B 0030282 11054 B 0030283 11056 B 0030284 11058 B 0030285 11060 B 0030286 11062 B 0030287 11063 B 0030288 11066 Mean S .D . Group 13 Gp Sex Age 13 13 13 13 13 13 13 13 CHOLEST MG/DL 58 62 66 49 60 56 65 76 61.5 8 GLUCOSE MG/DL 70 51 51 84 75 82 88 90 73.9 15.5 LDL-DIR MG/DL 39 37 54 38 45 36 51 63 45.4 9.8 HDL-DIR MG/DL 14 12 14 14 15 13 16 14 14 1.2 TRIG MG/DL 37 44 21 21 19 33 17 24 27 9.8 B 0030225 10915 B 0030226 10916 B 0030227 10917 B 0030228 10918 B 0030229 10919 B 0030230 10920 B 0030231 10925 B 0030232 10928 Mean S.D. Group 2 2 2 2 2 2 2 2 2 49 57 54 53 42 59 58 53 53.1 5.5 91 71 71 62 63 51 57 41 63.4 15 28 35 25 30 21 36 34 26 29.4 5.3 15 14 14 14 12 13 16 13 13.9 1.2 22 33 42 38 27 48 37 44 36.4 8.7 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 125 Page 2 + 000526 418-018:PAGE H-30 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Sgec. No. B 0030233 10930 B 0030234 10931 B 0030235 10932 B 0030236 10933 B 0030237 10934 B 0030238 10935 B 0030239 10937 B 0030240 10940 Mean S.D. Group 3 Gp Sex Age 3 3 3 3 3 3 3 3 CHOLEST MG/DL 71 66 64 58 100 63 63 61 68.3 13.4 GLUCOSE MG/DL 69 66 76 79 62 70 62 73 69.6 6.2 LDL-DIR MG/DL 48 49 48 44 67 46 41 43 48.3 8.1 HDL-DIR MG/DL 17 16 16 14 23 15 13 15 16.1 3 TRIG MG/DL 31 27 26 25 47 32 36 30 31.8 7.1 B 0030241 10945 B 0030242 10946 B 0030243 10947 B 0030244 10949 B 0030245 10950 B 0030246 10951 B 0030247 10953 B 0030248 10954 Mean S.D. Group 4 4 4 4 4 4 4 4 4 65 72 65 42 49 71 76 65 63.1 11.7 61 74 80 56 80 70 95 80 74.5 12.3 45 55 50 32 35 48 60 50 46.9 9.4 13 17 16 12 16 16 17 17 15.5 1.9 31 29 14 25 12 35 27 28 25.1 8.1 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 125 Page 3 + 000527 418-018:PAGE H-31 INCORPORATED 200 Girard Street, Suite 200. Gaithersburg. MD 20877 301-921-0168 Fax:301-977-0433 Client ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Spec. No. ID B 0030249 10958 B 0030250 10959 B 0030251 10960 B 0030252 10961 B 0030253 10962 B 0030254 10963 B 0030255 10964 B 0030256 10965 Mean S.D. Group 5 Gp Sex Age 5 5 5 5 5 5 5 5 CHOLEST MG/DL 54 40 44 41 45 47 44 46 45.1 4.3 GLUCOSE MG/DL 62 65 81 68 78 71 78 60 70.4 7.9 LDL-DIR MG/DL 30 18 20 21 29 29 27 19 24.1 5.1 HDL-DIR MG/DL 13 13 11 12 12 14 12 12 12.4 .9 TRIG MG/DL 34 39 35 27 21 25 29 44 31.8 7.6 B 0030257 10972 B 0030258 10974 B 0030259 10975 B 0030260 10976 B 0030261 10978 B 0030262 10980 B 0030263 10981 B 0030264 10982 Mean S.D. Group 6 6 6 6 6 6 6 6 6 45 64 82 59 47 54 66 56 59.1 11.8 99 57 50 99 81 58 92 83 77.4 19.8 39 46 49 44 36 39 51 47 43.9 5.4 14 16 14 16 14 11 15 15 14.4 1.6 10 26 55 26 16 29 25 16 25.4 13.6 Reference Range 48 96 74 319 Page 4 + 00 0 0 3125 000528 418-018:PAGE H-32 qYIICS 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NC OR P OR A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No., 1028 Study: 418018 PUPS GD21 Species: RAT Accession Spec. No. ID B 0030265 10985 B 0030266 10987 B 0030267 10988 B 0030268 10989 B 0030269 10990 B 0030270 10991 B 0030271 10992 B 0030272 10993 Mean S.D. Group 7 Gp Sex Age 7 7 7 7 7 7 7 7 CHOLEST MG/DL 64 85 67 59 65 49 65 58 64 10.3 GLUCOSE MG/DL 90 77 74 69 65 69 58 60 70.3 10.3 LDL-DIR MG/DL 47 62 49 45 45 35 43 39 45.6 8 HDL-DIR MG/DL 16 17 12 15 15 13 14 13 14.4 1.7 TRIG MG/DL 28 32 37 25 29 20 36 33 30 5.7 Reference Range 48 96 74 319 0- 0 00 Page 5 + 3125 000529 418-018-.PAGE H-33 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Sgec. No. B 0030217 10901 B 0030218 10902 B 0030219 10903 B 0030220 10904 B 0030221 10905 B 0030222 10907 B 0030223 10908 B 0030224 10910 Mean S.D. Group 1 Gp Sex Age 1 1 1 1 1 1 1 1 T4,TOTAL T3 ,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.69 0 .423 0 .845 TSH NG/ML FREE T4 NG/DL 0.05 .05 0 B 0030273 11040 B 0030274 11044 B 0030275 11045 B 0030276 11046 B 0030277 11047 B 0030278 11050 B 0030279 11051 B 0030280 11052 Mean S.D. Group 12 12 12 12 12 12 12 12 12 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.00 0 0 0.00 0.00 0.00 0 0 Reference Range 3 - 50 7 100 Page 6 + 00 .57 3.41 1.9 2.5 000530 418-018:PAGE H-34 c/ m Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Sgec. No. B 0030281 11053 B 0030282 11054 B 0030283 11056 B 0030284 11058 B 0030285 11060 B 0030286 11062 B 0030287 11063 B 0030288 11066 Mean S.D. Group 13 Gp Sex Age 13 13 13 13 13 13 13 13 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 1.16 1.59 0.00 2.80 0.00 0.00 0 .925 0 1.147 TSH NG/ML 1.16 1.16 0 FREE T4 NG/DL 0.00 0.00 0.00 0.00 0.00 0 0 B 0030225 10915 B 0030226 10916 B 0030227 10917 B 0030228 10918 B 0030229 10919 B 0030230 10920 B 0030231 10925 B 0030232 10928 Mean S.D. Group 2 2 2 2 2 2 2 2 2 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 8.43 0.00 0.00 2.81 4.867 0.07 .07 0 Reference Range 3 - 50 7 100 Page 7 + 0 0 .57 - 1.9 - 3.41 2.5 000531 418-018.-PAGE H-35 Qgyncsc V it 200 Girard Street, Suite 200. Gaithersburg, MD 20877 I N C OR P OR A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Spec. No. ID B 0030233 10930 B 0030234 10931 B 0030235 10932 B 0030236 10933 B 0030237 10934 B 0030238 10935 B 0030239 10937 B 0030240 10940 Mean S.D. Group 3 Gp Sex Age 3 3 3 3 3 3 3 3 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 2.19 0.00 2.70 0.00 2.78 0.00 0 1.096 0 0 1.379 0 TSH NG/ML FREE T4 NG/DL 0.00 0.00 0 0 B 0030241 10945 B 0030242 10946 B 0030243 10947 B 0030244 10949 B 0030245 10950 B 0030246 10951 B 0030247 10953 B 0030248 10954 Mean S.D. Group 4 4 4 4 4 4 4 4 4 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.00 5.10 1.33 6.68 0.38 4.08 2.928 2.747 0.00 0.00 0 0 0.00 0.00 0.00 0.00 0 0 Reference Range 3 - 50 7 100 Page 8 + 0 0 .57 - 1.9 - 3.41 2.5 000532 418-018.-PAGE H-36 i i JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443 -8710 Date Collected: Date Received: 12/08/2000 Date Reported: 01/16/2001 Client No . 1028 Study: 418018 PUPS GD21 Species: RAT Accession Spec. No. ID B 0030249 10958 B 0030250 10959 B 0030251 10960 B 0030252 10961 B 0030253 10962 B 0030254 10963 B 0030255 10964 B 0030256 10965 Mean S.D. Group 5 Gp Sex Age 5 5 5 5 5 5 5 5 T4 ,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 2.96 0.00 1.89 0.00 2.01 0.00 0.00 0 1.143 0 0 1.306 0 TSH NG/ML FREE T4 NG/DL 0.05 0.04 0.02 .037 .015 B 0030257 10972 B 0030258 10974 B 0030259 10975 B 0030260 10976 B 0030261 10978 B 0030262 10980 B 0030263 10981 B 0030264 10982 Mean S.D. Group 6 6 6 6 6 6 6 6 6 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.00 3.20 0.00 0.60 0.00 .76 1.389 0.00 0.00 0 0 0.04 .04 0 0.00 0.00 0.00 0.00 0 0 Reference Range 3 - 50 7 100 Page 9 + 00 .57 3.41 1.9 2.5 000533 418-018:PAGE H-37 Client INCORPORATED 200 Girard Street, Suite 200. Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/08/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Spec. No. ID B 0030265 10985 B 0030266 10987 B 0030267 10988 B 0030268 10989 B 0030269 10990 B 0030270 10991 B 0030271 10992 B 0030272 10993 Mean S.D. Group 7 Gp Sex Age 7 7 7 7 7 7 7 7 T4 ,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 1.96 0.00 0.00 0.00 0.00 0.00 0.00 0 .653 0 0 1.132 0 TSH NG/ML FREE T4 NG/DL 0.00 0.00 0 0 Reference Range 37 50 100 00 .57 - 1.9 - 3.41 2.5 Approved -- Page 10 of 10 D a t e -- 000534 418-018:PAGE H-38 c/m Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0030631 10906 B 0030632 10909 B 0030633 10911 B 0030634 10913 B 0030635 10914 Mean S.D. Group 1 Gp Sex Age 1 1 1 1 1 CHOLEST MG/DL 92 117 108 104 120 108.2 11.1 GLUCOSE MG/DL 292 229 188 276 250 247 40.9 LDL-DIR MG/DL 25 35 22 22 29 26.6 5.5 HDL-DIR MG/DL 26 38 25 28 18 27 7.2 TRIG MG/DL 203 243 213 153 235 209.4 35.4 B 0030673 11019 B 0030674 11021 B 0030675 11020 B 0030676 11023 B 0030677 11024 B 0030678 11025 B 0030679 11026. B 0030680 11027 B 0030681 11028 Mean S.D. Group 10 10 10 10 10 10 10 10 10 10 118 99 107 133 145 102 137 79 110 114.4 21 217 154 206 222 282 154 197 170 176 197.6 40.6 26 26 25 29 39 26 27 22 24 27.1 4.9 35 32 33 28 36 39 31 40 38 34.7 4 229 173 218 265 216 184 270 84 100 193.2 65.7 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 125 Page 1 + 000535 418-018:PAGE H-39 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: .01/16/2001 Client No,. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Spec. No. l5 B 0030682 11029 B 0030683 11031 B 0030684 11032 B 0030685 11033 B 0030686 11034 B 0030687 11035 B 0030688 11036 B 0030689 11037 B 0030690 11038 Mean S.D. Group 11 Gp Sex Age 11 11 11 11 11 11 11 11 11 CHOLEST MG/DL 131 108 126 120 131 198 100 135 107 128.4 28.8 GLUCOSE MG/DL 348 259 202 175 153 301 179 166 261 227.1 68.1 LDL-DIR MG/DL 32 32 33 21 26 80 30 17 32 33.7 18.3 HDL-DIR MG/DL 26 39 24 26 26 39 39 28 31 30.9 6.4 TRIG MG/DL 359 133 254 417 211 222 136 188 158 230.9 98.6 B 0030691 11049 Mean S.D. Group 12 12 192 192 0 245 245 0 77 77 0 40 40 0 212 212 0 B 0030692 11059 Mean S.D. Group 13 13 110 110 0 249 249 0 23 23 0 32 32 0 149 149 0 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 125 Page 2 + 000536 418-018:PAGE H-40 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: .01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Spec. No. if) B 0030636 10921 B 0030637 10922 B 0030638 10923 B 0030639 10924 B 0030640 10927 Mean S.D. Group 2 Gp Sex Age 2 2 2 2 2 CHOLEST MG/DL 128 128 118 119 107 120 8.7 GLUCOSE MG/DL 237 153 179 99 323 198.2 85.6 LDL-DIR MG/DL 31 28 47 38 27 34.2 8.3 HDL-DIR MG/DL 33 30 35 50 21 33.8 10.5 TRIG MG/DL 246 311 106 71 193 185.4 98.6 B 0030641 10929 B 0030642 10938 B 0030643 10942 Mean S.D. Group 3 3 3 3 148 202 151 167 30.3 203 267 394 288 97.2 47 80 29 52 25.9 42 30 30 34 6.9 232 371 333 312 71.8 B 0030644 10957 B 0030645 10966 B 0030646 10967 B 0030647 10968 B 0030648 10969 B 0030649 10970 Mean S.D. Group 5 5 5 5 5 5 5 125 130 112 117 124 104 118.7 9.6 277 382 155 289 262 244 268.2 73.3 23 20 22 28 20 1 19 9.3 28 20 21 30 30 19 24.7 5.2 215 789 149 285 139 681 376.3 284.8 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 125 Page 3 + 000537 418-018:PAGE H-41 <mcsc / m 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C OR P OR A T E D 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 01/16/2001 Client No,. 1028 Study : 418018 PUPS LD5 Species: RAT Accession Spec. No. ID B 0030650 10977 B 0030651 10979 B 0030652 10983 Mean S.D. Group 6 Gp Sex Age 6 6 6 CHOLEST MG/DL 173 118 128 139.7 29.3 GLUCOSE MG/DL 275 181 311 255.7 67.1 LDL-DIR MG/DL 66 36 30 44 19.3 HDL-DIR MG/DL 41 44 34 39.7 5.1 TRIG MG/DL 354 134 281 256.3 112.1 B 0030653 10995 B 0030654 10997 Mean S.D. Group 7 7 7 181 121 151 42.4 204 216 210 8.5 93 37 65 39.6 63 50 56.5 9.2 99 85 92 9.9 B 0030655 10999 B 0030656 11000 B 0030657 11001 B 0030658 11002 B 0030659 11003 B 0030660 11004 B 0030661 11005 B 0030662 11006 B 0030663 11007 Reference Range 8 8 8 8 8 8 8 8 8 114 109 101 123 139 121 118 104 98 48 96 319 290 177 375 190 240 228 190 174 74 319 17 21 30 20 37 26 30 31 28 00 25 26 50 21 31 28 34 39 40 00 366 342 100 444 269 230 235 127 124 3125 Page 4 + 000538 418-018:PAGE H-42 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0030664 11008 Mean S.D. Group 8 Gp Sex Age 8 CHOLEST MG/DL 122 114.9 12.4 GLUCOSE MG/DL 227 241 67.2 LDL-DIR MG/DL 37 27.7 6.8 HDL-DIR MG/DL 35 32.9 8.6 TRIG MG/DL 142 237.9 117.5 B 0030665 11009 B 0030666 11010 B 0030667 11011 B 0030668 11012 B 0030669 11013 B 0030670 11015 B 0030671 11016 B 0030672 11018 Mean S.D. Group 9 9 9 9 9 9 9 9 9 131 125 106 113 102 132 125 119 119.1 11.2 229 272 168 117 225 239 186 334 221.3 66.1 50 25 24 33 19 32 35 34 31.5 9.4 65 25 36 44 29 27 37 34 37.1 12.8 42 272 155 160 188 195 146 243 175.1 69.4 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 125 Page 5 + 000539 418-018:PAGE H-43 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/ 12/2000 Date Reported 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0030631 10906 B 0030632 10909 B 0030633 10911 B 0030634 10913 B 0030635 10914 Mean S.D. Group 1 Gp Sex Age 1 1 1 1 1 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.45 0.43 0.79 0.13 0.40 39.28 70.81 70.59 70.66 94.97 0.25 0.10 .44 .235 69.262 .175 19.786 .106 TSH NG/ML 0.76 .76 0 FREE T4 NG/DL 0.06 0.08 0.07 0.06 .068 .01 B 0030673 11019 B 0030674 11021 B 0030675 11020 B 0030676 11023 B 0030677 11024 B 0030678 11025 B 0030679 11026 B 0030680 11027 B 0030681 11028 Mean S.D. Group 10 10 10 10 10 10 10 10 10 10 0.00 0.00 0.00 0.00 0.00 0.15 0.00 0.00 0.00 .017 .05 51.23 40.88 56.43 50.60 41.97 57.95 34.91 47.71 8.613 0.00 0.00 0.00 0.00 0 0 2.36 2.36 0 0.01 0.01 0.01 0.01 0.03 0.03 0.01 .016 .01 Reference Range 3 - 50 7 100 Page 6 + 0 0 .57 3.41 1.9 2.5 00054# 418-018:PAGE H-44 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0030682 11029 B 0030683 11031 B 0030684 11032 B 0030685 11033 B 0030686 11034 B 0030687 11035 B 0030688 11036 B 0030689 11037 B 0030690 11038 Mean S.D. Group 11 Gp Sex Age 11 11 11 11 11 11 11 11 11 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 103.98 47.45 31.37 0.00 0.03 0.00 0.00 0.00 0.00 0.00 0.00 39.69 59.42 33.50 0.00 0.00 0 52.568 .006 0 27.185 .013 TSH NG/ML FREE T4 NG/DL 0.01 0.01 0.00 0.00 0.03 0.00 .008 .012 B 0030691 11049 Mean S.D. Group 12 12 0.00 0 0 42.35 42.35 0 0.02 .02 0 B 0030692 11059 Mean S.D. Group 13 13 0.00 0 0 40.17 40.17 0 0.15 .15 0 0.01 .01 0 Reference Range 3 - 50 7 100 Page 7 + 00 .57 3.41 1.9 2.5 000541 418-018:PAGE H-45 Client ociyVnZlcs INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0030636 10921 B 0030637 10922 B 0030638 10923 B 0030639 10924 B 0030640 10927 Mean S.D. Group 2 Gp Sex Age 2 2 2 2 2 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.80 0.21 0.61 0.37 0.44 92.83 67.70 134.10 52.98 69.49 0.51 0.98 0.00 .486 .227 83.42 .497 31.721 .49 TSH NG/ML 0.58 0.28 .43 .212 FREE T4 NG/DL 0.08 0.05 0.18 0.10 0.04 .09 .056 B 0030641 10929 B 0030642 10938 B 0030643 10942 Mean S.D. Group 3 3 3 3 0.00 0.00 0.00 0 0 58.69 54.04 99.17 70.633 24.823 0.68 .68 0 0.01 0.00 .005 .007 B 0030644 10957 B 0030645 10966 B 0030646 10967 B 0030647 10968 B 0030648 10969 B 0030649 10970 Mean S.D. Group 5 5 5 5 5 5 5 0.20 0.40 0.81 0.00 0.31 0.22 .323 .273 90.58 102.31 87.31 62.28 63.95 64.00 78.405 17.178 0.10 0.44 .27 .24 1.38 1.38 0 0.06 0.04 0.11 0.05 0.06 .064 .027 Reference Range 3 - 50 7 100 Page 8 + 00 .57 3.41 1.9 2.5 0C0542 418-018-.PAGE H-46 c S IN fi Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax; 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0030650 10977 B 0030651 10979 B 0030652 10983 Mean S.D. Group 6 Gp Sex Age 6 6 6 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 0.00 0.00 40.98 36.70 44.84 0.00 0 40.84 0 0 4.072 0 TSH NG/ML 0.00 0 0 FREE T4 NG/DL 0.00 0.00 0 0 B 0030653 10995 B 0030654 10997 Mean S.D. Group 7 7 7 0.00 0 0 25.90 25.9 0 B 0030655 10999 B 0030656 11000 B 0030657 11001 B 0030658 11002 B 0030659 11003 B 0030660 11004 B 0030661 11005 B 0030662 11006 B 0030663 11007 Reference Range 8 8 8 8 8 8 8 8 8 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 37 80.67 39.92 47.53 85.19 74.96 49.96 37.35 41.44 50 100 0.00 0.08 0.00 0.00 00 .57 3.41 0.01 0.03 0.01 0.01 0.02 0.02 0.01 1.9 2.5 Page 9 + 000543 418-018-.PAGEH-47 i jft JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 12/12/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0030664 11008 Mean S.D. Group 8 Gp Sex Age 8 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 0.00 44.97 0.00 0 55.777 .016 0. 18.934 .036 TSH NG/ML FREE T4 NG/DL 0.03 .018 .009 B 0030665 11009 B 0030666 11010 B 0030667 11011 B 0030668 11012 B 0030669 11013 B 0030670 11015 B 0030671 11016 B 0030672 11018 Mean S.D. Group 9 9 9 9 9 9 9 9 9 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 31.44 89.23 51.55 46.42 44.09 51.27 38.39 38.21 48.825 17.72 0.00 0.00 0.00 0.00 0.00 0 0 0.05 0.00 0.02 0.00 0.00 0.01 0.02 .014 .018 Reference Range 3- 50 - 0- .57 - 1.9 - 7 100 0 3.41 2.5 Approved --Page 10 of 10 Date - 000544 418-01S-.PAGE H-48 f vi INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 800-237-2815 OA AUDIT REPORT The report listed below was reviewed for compliance with the FDA Good Laboratory Practices and with EPA Good Laboratory Practices. The final report and all associated raw data were reviewed for accuracy and consistency and the findings were reported to management. The methods used were the methods described and the report accurately reflects the data. Therefore, these studies were done in compliance with the FDA and EPA Good Laboratory Practices. Franklin B. Newman QA Auditor SPONSOR: REPORT TYPE: Tt) fn , atcL. U \S STUDY: AUDIT DATE: REPORT TO MANAGEMENT: AUDIT #: I'W -o) Oh 0L3 DATE SUBMITTED TO CLIENT T c. L A <'7\y\ ri MANAGEMENT '/ w /of 000545 418-018-.PAGE H-49 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/01/2001 Date Reported: 04/17/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT ii i 1I 1 O1 1 TJ 1i Accession Sgec. No. B 0035529 11501 B 0035530 11502 B 0035531 11503 B 0035532 11504 B 0035533 11505 B 0035534 11506 B 0035535 11507 B 0035536 11508 B 0035537 11509 B 0035538 11510 B 0035539 11511 B 0035540 11512 B 0035541 11513 B 0035542 11514 Mean S.D. Group 1 Sex Age CHOLEST MG/DL 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 165 158 169 99 135 126 104 130 97 116^ 129 98 92 135 125.2 25.8 B 0035609 11620 B 0035610 11621 B 0035611 11622 B 0035612 11623 B 0035613 11624 B 0035614 11625 B 0035615 11626 Reference Range 10 F 10 F 10 F 10 F 10 F 10 F 10 F 78 112 130 103 78 78 164 48 96 Page 1 + 000546 418-018-.PAGE H-50 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/01/2001 Date Reported: 04/17/2001 Client No., 1028 Study: 418018 LD5 MILK Species: RAT Accession Sgec. No. B 0035616 11628 B 0035617 11629 Mean S.D. Group 10 Gp Sex Age 10 F 10 F CHOLEST MG/DL 84 111 104.2 29.2 B 0035618 11630 B 0035619 11632 B 0035620 11633 B 0035621 11634 B 0035622 11635 B 0035623 11636 B 0035624 11637 B 0035625 11638 B 0035626 11639 Mean S.D. Group 11 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 86 82 70 77 87 129 67 75 91 84.9 18.4 B 0035627 11640 B 0035628 11641 B 0035629 11642 Reference Range 12 F 12 F 12 F 93 57 80 48 96 Page 2 + 000547 418-018:PAGE H-51 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/01/2001 Date Reported: 04/17/2001 Client No.. 1028 Study: 418018 LD5 MILK Species: RAT Accession Spec. No. ID B 0035630 11643 B 0035631 11644 B 0035632 11646 B 0035633 11647 B 0035634 11648 B 0035635 11650 B 0035636 11651 B 0035637 11653 Mean S.D. Group 12 Gp Sex Age 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F CHOLEST MG/DL 106 76 89 25 93 59 69 59 73.3 22.7 B 0035638 11654 B 0035639 11655 B 0035640 11656 B 0035641 11662 Mean S.D. Group 13 13 F 13 F 13 F 13 F 92 98 84 96 92.5 6.2 B 0035543 11515 B 0035544 11516 Reference Range 2F 2F 89 48 96 Page 3 + 000548 418-018:PAGE H-52 Client INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/01/2001 Date Reported: 04/17/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Sgec. No. B 0035545 11517 B 0035546 11518 B 0035547 11519 B 0035548 11520 B 0035549 11521 B 0035550 11523 B 0035551 11524 B 0035552 11525 B 0035553 11526 B 0035554 11527 B 0035555 11528 Mean S.D. Group 2 Gp Sex Age 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F CHOLEST MG/DL 118 97 97 87 103 109 130 116 120 111 167 112 21.7 B 0035556 11529 B 0035557 11530 B 0035558 11534 B 0035559 11536 B 0035560 11537 B 0035561 11538 B 0035562 11540 B 0035563 11542 B 0035564 11543 Mean S.D. Group 3 3F 3F 3F 3F 3F 3F 3F 3F 3F 77 96 62 81 109 42 121 136 103 91.9 29.5 Reference Range 48 96 Page 4 + 000549 418-018:PAGE H-53 V t A L -J l A. JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/01/2001 Date Reported: . 04/17/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Spec. No. ID B 0035565 11547 B 0035566 11552 Mean S.D. Group 4 Gp Sex Age 4F 4F CHOLEST MG/DL 269 107 188 114.6 B 0035567 11558 B 0035568 11559 B 0035569 11561 B 0035570 11563 B 0035571 11564 B 0035572 11565 B 0035573 11567 B 0035574 11568 B 0035575 11569 B 0035576 11570 B 0035577 11571 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 5F 5F 5F 5F 5F 109 94 136 137 104 149 100 99 92 187 88 117.7 30.9 B 0035578 11574 Reference Range 6F 72 48 96 Page 5 + 000550 418-018:PAGE H-54 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORPORATED 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/01/2001 Date Reported: 04/17/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Sgec. No. B 0035579 11575 B 0035580 11576 B 0035581 11578 B 0035582 11579 B 0035583 11580 B 0035584 11582 B 0035585 11583 B 0035586 11584 Mean S.D. Group 6 ft l Ol l l 1 Sex Age CHOLEST MG/DL 6F 6F 6F 6F 6F 6F 6F 6F 127 138 68 116 105 149 93 102 107.8 27.8 B 0035587 11592 B 0035588 11593 B 0035589 11597 Mean S.D. Group 7 7F 7F 7F 47 76 61.5 20.5 B 0035590 11600 B 0035591 11601 B 0035592 11602 Reference Range 8F 8F 8F 134 79 74 48 96 Page 6 + 000551 418-018:PAGE H-55 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORPORATED 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/01/2001 Date Reported: 04/17/2001 Client No. 1028 Study: 418018 LD5 MILK Species: RAT Accession Spec. No. ID B 0035593 11603 B 0035594 11604 B 0035595 11605 B 0035596 11606 B 0035597 11607 B 0035598 11608 B 0035599 11609 Mean S.D. Group 8 Gp Sex Age 8F 8F 8F 8F 8F 8F 8F CHOLEST MG/DL 89 146 106 226 126 155 103 123.8 45.3 B 0035600 11611 B 0035601 11612 B 0035602 11613 B 0035603 11614 B 0035604 11615 B 0035605 11616 B 0035606 11617 B 0035607 11618 B 0035608 11619 Mean S.D. Group 9 9F 9F 9F 9F 9F 9F 9F 9F 9F 97 98 139 141 77 92 96 100 135 108.3 23.5 Reference Range 48 96 Approved -----Page 7 '"of D a te 000552 418-018:PAGE H-56 4 I /I JL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No,. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Spec. No. ID B 0035787 11501 B 0035788 11502 B 0035789 11503 B 0035790 11504 B 0035791 11505 B 0035792 11506 B 0035793 11507 B 0035794 11508 B 0035795 11509 B 0035796 11510 B 0035797 11511 B 0035798 11512 B 0035799 11513 B 0035800 11514 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F CHOLEST MG/DL 96 86 67 100 64 85 91 97 95 59 53 84 69 62 79.1 16.2 GLUCOSE MG/DL 207 132 162 206 204 145 174 154 168 155 149 209 177 203 174.6 26.7 LDL-DIR MG/DL 7 10 7 7 5 6 4 7 5 1 1 6 3 2 5.1 2.6 HDL-DIR MG/DL 81 73 53 83 60 70 73 85 85 51 45 74 50 48 66.5 14.8 TRIG MG/DL 133 82 125 83 71 107 183 92 112 137 149 112 173 173 123.7 36.2 B 0035801 11630 B 0035802 11632 B 0035803 11633 B 0035804 11634 B 0035805 11635 B 0035806 11636 B 0035807 11637 Reference Range 11 F 11 F 11 F 11 F 11 F 11 F 11 F 44 78 66 79 49 74 63 48 96 200 145 208 161 166 156 180 74 319 5 1 4 6 1 6 3 00 39 67 62 74 42 70 61 20 80 102 170 129 107 157 93 136 3104 Page l + 000553 418-018:PAGE H-57 M j l *dl A. X V V J 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C OR P O R A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No . 1028 Study: 418018 DAMS LD5 Species: RAT 1 0. 1o 1 11 Accession Spec. No. ID B 0035808 11638 B 0035809 11639 Mean S.D. Group 11 Sex Age 11 F 11 F CHOLEST MG/DL 69 69 65.7 12.1 GLUCOSE MG/DL 179 185 175.6 20.6 LDL-DIR MG/DL 6 4 4 2 HDL-DIR MG/DL 62 67 60.4 12.1 TRIG MG/DL 98 90 120.2 29.2 B 0035810 11640 B 0035811 11641 B 0035812 11642 B 0035813 11643 B 0035814 11644 B 0035815 11645 B 0035816 11646 B 0035817 11647 B 0035818 11648 B 0035819 11649 B 0035820 11650 B 0035821 11651 B 0035822 11653 Mean S.D. Group 12 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 61 43 57 39 46 77 52 41 68 78 50 47 63 55.5 13.1 179 186 165 190 180 175 157 207 145 206 159 176 159 175.7 18.7 2 1 7 3 3 2 2 2 2 8 2 1 7 3.2 2.4 54 43 51 37 38 59 51 40 66 69 48 46 58 50.8 10.2 183 77 63 68 153 190 62 44 77 62 54 56 60 88.4 51 Reference Range 48 96 74 319 Page 2 + 00 20 80 3104 000554 418-018:PAGE H-58 \j*2JL^LL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0035823 11654 B 0035824 11655 B 0035825 11656 B 0035826 11657 B 0035827 11658 B 0035828 11659 B 0035829 11660 B 0035830 11661 B 0035831 11662 B 0035832 11663 B 0035833 11664 B 0035834 11665 B 0035835 11667 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F CHOLEST MG/DL 44 59 50 64 52 61 57 69 37 58 77 60 67 58.1 10.6 GLUCOSE MG/DL 208 168 175 222 206 158 196 197 202 153 152 227 202 189.7 25.6 LDL-DIR MG/DL 4 4 3 6 7 2 5 5 2 3 5 7 5 4.5 1.7 HDL-DIR MG/DL 39 56 45 60 46 60 54 65 36 51 61 53 61 52.8 9 TRIG MG/DL 125 48 122 74 50 99 66 93 20 118 159 54 126 88.8 40.4 Reference Range 48 96 74 319 Page 3 + 00 20 80 3104 0G0555 418-018-.PAGE H-59 Client 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORPORATED 301-921-0168 Fax:301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0035787 11501 B 0035788 11502 B 0035789 11503 B 0035790 11504 B 0035791 11505 B 0035792 11506 B 0035793 11507 B 0035794 11508 B 0035795 11509 B 0035796 11510 B 0035797 11511 B 0035798 11512 B 0035799 11513 B 0035800 11514 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F T3,TOTAL T 4 ,TOTAL TSH NG/DL UG/DL NG/ML 65.64 104.47 37.77 73.00 55.01 63.16 57.61 76.31 79.83 71.86 58.43 80.84 53.33 65.08 1.30 2.97 0.85 1.11 1.35 1.28 1.08 2.39 1.49 0.88 0.68 1.25 0.95 0.92 1.68 1.55 0.91 2.95 1.67 1.19 3.24 2.62 1.42 0.77 1.43 2.03 0.91 0.66 67.31 1.321 15.865 .628 1.645 .808 FREE T3 PG/ML 0.43 1.19 0.04 0.58 0.50 0.47 0.15 0.59 0.89 0.56 0.54 0.69 0.39 0.62 .546 .28 FREE T4 NG/DL 0.62 1.42 0.55 0.78 0.77 0.70 0.64 1.03 0.68 0.79 0.48 0.92 0.80 0.77 .782 .232 B 0035801 11630 B 0035802 11632 B 0035803 11633 B 0035804 11634 B 0035805 11635 B 0035806 11636 B 0035807 11637 Reference Range 11 F 11 F 11 F 11 F 11 F 11 F 11 F 26.82 47.75 37.99 46.74 46.41 43.09 48.31 50 100 0.37 0.06 0.00 0.01 0.06 0.05 0.11 37 2.54 0.50 1.66 0.98 1.52 1.58 4.35 .57 3.41 0.18 0.55 0.11 0.15 0.18 0.26 0.35 00 0.28 0.20 0.17 0.29 0.28 0.26 0.21 1.9 2.5 Page 4 + 0C055S 418-018:PAGE H-60 c yiNfi 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORPORATED 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 DAMS LD5 Species : RAT Accession Sgec. No. B 0035808 11638 B 0035809 11639 Mean S.D. Group 11 Gp Sex Age 11 F 11 F T3,TOTAL T4 ,TOTAL TSH NG/DL UG/DL NG/ML 31.70 51.74 0.10 0.13 0.60 1.64 42.283 .099 8.387 .111 1.708 1.17 FREE T3 PG/ML 0.09 0.36 .248 .149 FREE T4 NG/DL 0.18 0.29 .24 .049 B 0035810 11640 B 0035811 11641 B 0035812 11642 B 0035813 11643 B 0035814 11644 B 0035815 11645 B 0035816 11646 B 0035817 11647 B 0035818 11648 B 0035819 11649 B 0035820 11650 B 0035821 11651 B 0035822 11653 Mean S.D. Group 12 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 36.31 32.93 32.46 41.21 26.20 54.83 28.42 55.92 33.95 59.37 31.35 29.13 43.24 38.871 11.249 0.00 0.20 0.08 0.24 0.08 0.29 0.38 0.63 0.16 0.26 0.29 0.31 0.23 .242 .158 1.13 2.25 3.11 1.25 1.68 1.30 1.46 1.48 1.79 2.38 3.38 0.46 0.71 1.722 .861 0.03 0.12 0.00 0.34 0.02 0.31 0.29 0.50 0.00 0.57 0.14 0.06 0.00 .183 .199 0.19 0.23 0.14 0.29 0.19 0.21 0.20 0.41 0.21 0.31 0.21 0.30 0.32 .247 .074 Reference Range 50 100 37 Page 5 + .57 3.41 00 1.9 2 .!> 0C0557 418-018:PAGE H-61 % M, j i J!t JL JL V->rVw/ 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C OR P O R A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No., 1028 Study: 418018 DAMS LD5 Species : RAT Accession Sgec. No. B 0035823 11654 B 0035824 11655 B 0035825 11656 B 0035826 11657 B 0035827 11658 B 0035828 11659 B 0035829 11660 B 0035830 11661 B 0035831 11662 B 0035832 11663 B 0035833 11664 B 0035834 11665 B 0035835 11667 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F T3 ,TOTAL T4 ,TOTAL TSH NG/DL UG/DL NG/ML 35.84 39.95 28.99 46.76 46.50 33.35 48.66 59.68 21.45 56.44 77.52 52.57 54.46 0.25 0.49 0.10 0.18 0.11 0.22 0.23 0.38 0.32 0.06 0.18 0.20 0.08 1.70 1.45 1.27 1.51 1.18 2.23 1.03 2.19 0.93 0.91 1.76 0.74 1.49 46.321 .215 14.728 .124 1.415 .47 FREE T3 PG/ML 0.14 0.24 0.00 0.41 0.24 0.01 0.75 0.47 0.01 0.38 0.53 0.36 0.35 .299 .224 FREE T4 NG/DL 0.24 0.33 0.17 0.36 0.23 0.30 0.26 0.37 0.20 0.21 0.25 0.35 0.40 .282 .074 Reference Range 50 100 37 .57 3.41 00 1.9 2.5 Approved -- Page 6 of 6 Date 000558 418-018:PAGE H-62 M A L - J i J. JLV-yV-y 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NC OR P OR A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No., 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0035934 11620 B 0035935 11621 B 0035936 11622 B 0035937 11623 B 0035938 11624 B 0035939 11625 B 0035940 11626 B 0035941 11627 B 0035942 11628 B 0035943 11629 Mean S.D. Group 10 Gp Sex Age 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F CHOLEST MG/DL 44 73 67 46 80 67 48 55 72 50 60.2 13 GLUCOSE MG/DL 197 172 155 151 175 149 156 188 170 171 168.4 15.9 LDL-DIR MG/DL 2 8 5 3 3 4 4 4 3 1 3.7 1.9 HDL-DIR MG/DL 43 66 59 44 67 62 45 50 66 48 55 9.9 TRIG MG/DL 115 90 122 47 123 47 91 84 116 154 98.9 34.1 B 0035836 11515 B 0035837 11516 B 0035838 11517 B 0035839 11518 B 0035840 11519 B 0035841 11520 B 0035842 11521 B 0035843 11523 B 0035844 11524 B 0035845 11525 B 0035846 11526 Reference Range 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 86 101 66 83 94 88 65 105 64 89 89 48 96 196 168 157 168 210 162 279 157 203 243 178 74 319 3 5 1 4 9 4 6 4 3 4 5 00 69 96 62 78 82 61 60 81 61 82 71 20 80 148 48 113 127 94 179 90 149 114 133 157 3104 Page 1 + 000559 418-018:PAGE H-63 Client omcsc / m m , 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORPORATED 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No,. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0035847 11527 B 0035848 11528 Mean S.D. Group 2 Gp Sex Age 2F 2F CHOLEST MG/DL 69 80 83 13.6 GLUCOSE MG/DL 137 148 185.1 40.5 LDL-DIR MG/DL 1 3 4 2.1 HDL-DIR MG/DL 50 78 71.6 12.6 TRIG MG/DL 293 72 132.1 60.5 B 0035849 11529 B 0035850 11530 B 0035851 11532 B 0035852 11533 B 0035853 11534 B 0035854 11535 B 0035855 11536 B 0035856 11537 B 0035857 11538 B 0035858 11539 B 0035859 11540 B 0035860 11541 B 0035861 11542 B 0035862 11543 Mean S.D. Group 3 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 67 67 79 74 55 68 59 60 51 56 106 77 53 72 67.4 14.3 225 166 194 184 188 163 185 144 198 210 155 172 182 189 182.5 21.5 5 5 2 4 4 1 2 5 1 5 8 6 3 3 3.9 2 63 60 63 67 52 56 58 55 52 44 79 60 42 69 58.6 9.7 103 113 184 88 48 201 67 109 80 89 46 110 130 58 101.9 46.1 Reference Range 48 96 74 319 Page 2 + 00 20 80 3104 000560 418-018:PAGE H-64 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORPORATED 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No . 1028 Study: 418018 DAMS LD5 Species: RAT Accession Spec. No. ID B 0035863 11544 B 0035864 11545 B 0035865 11546 B 0035866 11547 B 0035867 11549 B 0035868 11550 B 0035869 11551 B 0035870 11552 B 0035871 11553 B 0035872 11554 B 0035873 11555 B 0035874 11556 B 0035875 11557 Mean S.D. Group 4 Gp Sex Age 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F CHOLEST MG/DL 69 62 79 50 73 55 82 44 89 62 68 72 64 66.8 12.7 GLUCOSE MG/DL 163 178 186 254 191 247 180 148 211 193 184 170 171 190.5 30.7 LDL-DIR MG/DL 2 1 2 2 1 6 8 1 5 4 5 2 2 3.2 2.2 HDL-DIR MG/DL 44 55 59 48 72 51 75 39 68 55 62 62 44 56.5 11.2 TRIG MG/DL 211 158 139 41 84 65 71 135 169 113 78 138 190 122.5 52.1 B 0035876 11558 B 0035877 11559 B 0035878 11561 B 0035879 11563 B 0035880 11564 B 0035881 11565 B 0035882 11567 B 0035883 11568 Reference Range 5F 5F 5F 5F 5F 5F 5F 5F 91 91 88 62 59 71 75 84 48 96 154 136 159 178 189 186 220 172 74 319 1 3 7 6 7 5 4 2 00 66 70 79 54 49 65 55 69 20 80 189 139 84 100 101 96 147 136 3104 Page 3 + 000561 418-018:PAGE H-65 csrnm Q x n cs INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No.. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sgec. No. B 0035884 11569 B 0035885 11570 B 0035886 11571 Mean S.D. Group 5 Gp Sex Age 5F 5F 5F CHOLEST MG/DL 108 53 112 81.3 19.3 GLUCOSE MG/DL 189 234 174 181 28 LDL-DIR MG/DL 9 3 8 5 2.6 HDL-DIR MG/DL 90 51 78 66 13 TRIG MG/DL 133 69 169 123.9 37.1 B 0035887 11572 B 0035888 11573 B 0035889 11574 B 0035890 11575 B 0035891 11576 B 0035892 11577 B 0035893 11578 B 0035894 11579 B 0035895 11580 B 0035896 11581 B 0035897 11582 B 0035898 11583 B 0035899 11584 B 0035900 11585 Mean S.D. Group 6 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 56 82 49 57 57 63 52 47 46 71 37 62 54 49 55.9 11.3 194 192 175 188 137 170 177 192 157 181 164 172 143 185 173.4 17.9 6 5 6 5 6 2 3 8 1 2 2 6 2 6 4.3 2.2 52 78 45 53 50 51 44 40 43 68 37 56 53 43 50.9 11 53 92 60 92 89 148 110 57 103 153 46 89 63 69 87.4 33.1 Reference Range 48 96 74 319 Page 4 + 00 20 80 3104 0005S2 418-018:PAGE H-66 \ L a J~JLL JL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Spec. No. ID B 0035901 11586 B 0035902 11587 B 0035903 11588 B 0035904 11589 B 0035905 11590 B 0035906 11591 B 0035907 11592 B 0035908 11593 B 0035909 11594 B 0035910 11595 B 0035911 11596 B 0035912 11597 B 0035913 11598 B 0035914 11599 Mean S.D. Group 7 B 0035915 11600 B 0035916 11601 B 0035917 11602 B 0035918 11603 B 0035919 11604 B 0035920 11605 B 0035921 11606 Reference Range Gp Sex Age 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F CHOLEST MG/DL 88 69 68 41 57 74 52 38 63 44 67 38 69 62 59.3 15 GLUCOSE MG/DL 194 188 182 206 173 152 192 256 248 163 181 198 203 217 196.6 29 LDL-DIR MG/DL 11 5 4 1 7 7 2 4 6 3 4 5 6 4 4.9 2.5 HDL-DIR MG/DL 79 66 60 39 47 57 51 31 58 41 65 35 63 54 53.3 13.5 TRIG MG/DL 86 67 135 31 104 105 75 130 54 51 43 79 87 177 87.4 40.1 8F 8F 8F 8F 8F 8F 8F 100 81 44 67 64 61 48 48 96 167 180 144 216 190 200 157 74 319 5 3 4 5 5 3 3 00 96 71 40 64 60 57 46 20 80 58 220 67 104 60 81 58 3104 Page 5 + 090563 418-018:PAGE H-67 c/ m INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received; 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 DAMS LD5 Species : RAT Accession Spec. No. ID B 0035922 11607 B 0035923 11608 B 0035924 11609 Mean S.D. Group 8 Gp Sex Age 8F 8F 8F CHOLEST MG/DL 79 68 74 68.6 16.3 GLUCOSE MG/DL 212 165 172 180.3 23.8 LDL-DIR MG/DL 9 5 5 4.7 1.8 HDL-DIR MG/DL 70 62 70 63.6 15.3 TRIG MG/DL 56 63 90 85.7 49.8 B 0035925 11611 B 0035926 11612 B 0035927 11613 B 0035928 11614 B 0035929 11615 B 0035930 11616 B 0035931 11617 B 0035932 11618 B 0035933 11619 Mean S.D. Group 9 9F 9F 9F 9F 9F 9F 9F 9F 9F 59 44 72 87 77 43 61 72 46 62.3 15.8 182 162 182 145 187 162 154 181 176 170.1 14.8 6 2 4 4 7 2 4 3 2 3.8 1.8 55 44 69 71 62 37 58 70 46 56.9 12.4 118 99 48 147 167 148 83 55 66 103.4 43.9 Reference Range 48 96 74 319 00 Approved -Page 6 of 6 20 80 - Date 3104 0G05G4 418-018:PAGE H-68 <^/2TvH (syncs INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/ 02/2001 Date Reported: .02/13/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. Gp Sex Age B 0035728 11501/506 1 B 0035729 11502/507 1 B 0035730 11504 1 B 0035731 11508/503 1 B 0035732 11509/510 1 B 0035733 11512/511 1 B 0035734 11513/514 1 Mean S.D. Group 1 CHOLEST MG/DL 110 111 107 111 136 115 119 115.6 9.8 GLUCOSE MG/DL 327 312 320 314 245 148 169 262.1 76.1 LDL-DIR MG/DL 32 35 36 30 41 47 48 38.4 7.1 HDL-DIR MG/DL 24 29 28 25 29 42 45 31.7 8.3 TRIG MG/DL 235 212 298 241 271 134 133 217.7 63.7 B 0035735 11630/633 11 B 0035736 11632/636 11 B 0035737 11635/637 11 B 0035738 11638 11 B 0035739 11639/634 11 Mean S.D. Group 11 114 115 120 125 116 118 4.5 141 267 201 201 187 199.4 45.1 28 37 48 32 39 36.8 7.6 28 38 38 30 35 33.8 4.6 242 207 218 266 213 229.2 24.5 B 0035740 11640/641 12 B 0035741 11646/644 12 Reference Range 132 118 48 96 217 184 74 319 29 30 00 26 37 20 80 263 276 3125 Page 1 + 000565 418-018:PAGE H-69 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/ 02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Spec. No. ID Gp Sex Age B 0035742 11648/65112 B 0035743 11650 12 B 0035744 11653/642 12 Mean S.D. Group 12 CHOLEST MG/DL 145 95 112 120.4 19.1 GLUCOSE MG/DL 227 233 226 217.4 19.5 LDL-DIR MG/DL 50 23 33 33 10.2 HDL-DIR MG/DL 36 31 46 35.2 7.5 TRIG MG/DL 292 163 185 235.8 57.9 B 0035745 11654 13 B 0035746 11656/655 13 Mean S.D. Group 13 172 102 137 49.5 217 222 219.5 3.5 81 30 55.5 36.1 39 32 35.5 4.9 255 235 245 14.1 Reference Range 48 96 74 319 Page 2 + 0 0 20 80 3125 000566 418-018:PAGE H-70 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession |gec. No. Gp Sex Age B 0035728 11501/506 1 B 0035729 11502/507 1 B 0035730 11504 1 B 0035731 11508/503 1 B 0035732 11509/510 1 B 0035733 11512/511 1 B 0035734 11513/514 1 Mean S.D. Group 1 T3,TOTAL T4 ,TOTAL TSH NG/DL UG/DL NG/ML 38.73 48.84 45.82 44.65 46.29 43.06 38.90 0.60 0.65 0.97 0.54 0.67 0.57 0.32 0.88 1.17 1.27 0.99 1.10 1.06 0.91 43.756 .617 3.8 .194 1.054 .14 FREE T3 PG/ML 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 FREE T4 NG/DL 0.11 0.12 0.12 0.11 0.11 0.08 0.07 .103 .02 B 0035735 11630/633 11 B 0035736 11632/636 11 B 0035737 11635/637 11 B 0035738 11638 11 B 0035739 11639/634 11 Mean S.D. Group 11 35.45 33.85 32.59 47.34 27.23 35.292 7.409 0.00 0.05 0.00 0.00 0.02 .014 .022 1.09 1.07 1.35 0.79 0.75 1.01 .246 0.00 0.00 0.00 0.00 0.00 0 0 0.01 0.02 0.01 0.01 0.02 .014 .005 B 0035740 11640/641 12 B 0035741 11646/644 12 Reference Range 35.15 36.35 50 100 0.00 0.03 37 1.91 1.15 .57 3.41 0.02 0.00 00 0.01 0.01 1.9 2.5 Page 3 + 000567 418-018:PAGE H-71 z J lN I' INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/ 02/2001 Date Reported 02/13/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. Gp Sex Age B 0035742 11648/651 12 B 0035743 11650 12 B 0035744 11653/642 12 Mean S.D. Group 12 T3,TOTAL T4,TOTAL TSH NG/DL UG/DL NG/ML 33.69 18.76 29.95 0.00 0.00 0.00 1.62 1.48 1.09 30.78 7.137 .006 .013 1.45 .339 FREE T3 PG/ML 0.00 0.00 0.00 .004 .009 FREE T4 NG/DL 0.01 0.00 0.01 .008 .004 B 0035745 11654 13 B 0035746 11656/655 13 Mean S.D. Group 13 18.74 39.42 29.08 14.623 0.00 0.00 0 0 1.50 1.5 0 0.00 0.00 0 0 0.00 0.02 .01 .014 Reference Range 50 100 37 .57 3.41 00 1.9 2.5 A p p r o v e d-- -- Page 4 of Date 000568 418-018:PAGE H-72 CLIENT: SPECIMEN ID: SPECIES: STUDY: GROUP: ACCESSION: PROFILE/TEST ANI LYTICS, INC. 200 GIRARD STREET SUITE 200 GAITHERSBURG, MD 20877 (301) 921-0168 or (FAX) 977-0433 ARGUS RESEARCH LABORATORIES, 11654 RAT 418018 PUPS LD5 13 B 0035745 INCDATE SPEC. COLLECTED: DATE SPEC. RECEIVED: DATE REPORTED: AGE: SEX: 02/ 02/2 02/13/2 RESULTS UNITS REFERENCE: RANGE ARGUS CHEM PROFILE CHOLESTEROL GLUCOSE LDL-DIRECT HDL-DIRECT TRIGLYCERIDE TRIIODOTHYRONINE THYROXINE TSH, RAT T3, FREE FREE T4 172 217 81 39 255 18.74 0.00 QNS 0.00 0.00 MG/DL MG/DL MG/DL MG/DL MG/DL NG/DL UG/DL NG/ML PG/ML NG/DL 48 - 96 74 - 319 20 - 80 3 - 125 50 - 100 3.0 - 7.0 0.57 - 3.41 1.9 - 2.5 Laboratory Director: Saroj R. Das, Ph.D. PAGE 1 OF 1 FINAL REPO! i 0C059 418-018:PAGE H-73 . /I JL 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C OR P O R A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No.. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Spec. No. ID Gp Sex Age B 0035782 11620/626 10 B 0035783 11621/622 10 B 0035784 11623 10 B 0035785 11624/629 10 B 0035786 11625/628 10 Mean S.D. Group 10 CHOLEST MG/DL 115 108 106 108 117 110.8 4.9 GLUCOSE MG/DL 173 255 290 177 267 232.4 53.9 LDL-DIR MG/DL 37 34 28 32 36 33.4 3.6 HDL-DIR MG/DL 48 43 27 31 47 39.2 9.6 TRIG MG/DL 135 169 258 298 170 206 68.7 B 0035747 11515/524 2 B 0035748 11517/520 2 B 0035749 11519/518 2 B 0035750 11521/516 2 B 0035751 11523/527 2 B 0035752 11525/528 2 B 0035753 11526 2 Mean S.D. Group 2 111 111 110 121 117 109 106 112.1 5.1 213 357 238 327 152 244 375 272.3 82.3 42 31 30 37 49 28 36 36.1 7.4 41 31 44 30 50 31 29 36.6 8.3 158 225 173 294 152 188 202 198.9 49 B 0035754 11536/537 3 B 0035755 11538/534 3 Reference Range 149 174 48 96 217 310 74 319 53 75 00 32 37 20 80 247 329 3125 Page 1 + OC0570 418-018:PAGE H-74 i t .Jl JL J L K J l + J 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I N C O R P O R A T E D 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No,. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Spec. No. ID Gp Sex Age B 0035756 11540/530 3 B 0035757 11543/529 3 Mean S.D. Group 3 CHOLEST MG/DL 145 130 149.5 18.3 GLUCOSE MG/DL 284 238 262.3 42.4 LDL-DIR MG/DL 40 32 50 18.8 HDL-DIR MG/DL 32 28 32.3 3.7 TRIG MG/DL 234 370 295 65.3 B 0035758 11558/569 5 B 0035759 11559/570 5 B 0035760 11561/568 5 B 0035761 11563/564 5 B 0035762 11565/567 5 B 0035763 11571 5 Mean S.D. Group 5 122 102 111 119 118 129 116.8 9.3 325 288 253 237 223 230 259.3 39.6 24 41 37 21 41 44 34.7 9.7 24 39 32 22 39 40 32.7 8 289 145 295 370 178 321 266.3 86.7 B 0035764 11574/575 6 B 0035765 11576/578 6 B 0035766 11582 6 B 0035767 11583/580 6 B 0035768 11584/579 6 Mean S.D. Group 6 131 117 125 132 116 124.2 7.5 227 257 303 243 185 243 43.1 47 40 37 40 32 39.2 5.4 40 44 37 38 36 39 3.2 193 147 286 243 225 218.8 52.3 Reference Range 48 96 74 319 Page 2 + 00 20 80 3125 000571 418-018:PAGE H-75 c/J N L INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM. PA 19044(215) 443-8710 Date Collected: Date Received: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Sgec. No. B 0035769 11592 B 0035770 11593 B 0035771 11597 Mean S.D. Group 7 Gp Sex Age 7 7 7 CHOLEST MG/DL 130 121 121 124 5.2 GLUCOSE MG/DL 255 211 230 232 22.1 LDL-DIR MG/DL 42 46 58 48.7 8.3 HDL-DIR MG/DL 28 44 51 41 11.8 TRIG MG/DL 160 160 98 139.3 35.8 B 0035772 11600/608 8 B 0035773 11602/609 8 B 0035774 11603/604 8 B 0035775 11606/601 8 B 0035776 11607/605 8 Mean S.D. Group 8 106 97 114 104 109 106 6.3 307 244 286 179 242 251.6 49.2 35 18 31 34 41 31.8 8.5 33 33 31 31 34 32.4 1.3 249 228 195 248 225 229 22 B 0035777 11611 9 B 0035778 11612/613 9 B 0035779 11614/616 9 B 0035780 11617/619 9 B 0035781 11618/615 9 Mean S.D. Group 9 100 120 108 106 106 108 7.3 194 173 284 233 210 218.8 42.6 28 37 33 26 33 31.4 4.4 46 46 30 45 31 39.6 8.3 121 120 233 178 276 185.6 68.8 Reference Range 48 96 74 - 0- 319 0 Approved -- -- Page 3 of 3 20 80 Date -- 3125 OC0572 418-018.-PAGE H-76 I vf J L \**A *J INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, M D 20877 301-921-0168 800-237-2815 OA AUDIT REPORT The report listed below was reviewed for compliance with the FDA Good Laboratory Practices and with EPA Good Laboratory Practices. The final report and all associated raw data were reviewed for accuracy and consistency and the findings were reported to management. The methods used were the methods described and the report accurately reflects the data. Therefore, these studies were done in compliance with the FDA and EPA Good Laboratory Practices. Franklin B. Newman QA Auditor SPONSOR: /jft-ts' ABZ STUDY: / j ) $ i i r f REPORT TYPE: O T 3 , TV, T5tt rf& T3 j ffZT AUDIT DATE: REPORT TO MANAGEMENT: AUDIT #: &-)<?' o I OC 0573 418-018:PAGE H-77 l JL JLV-/VJ INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 800-237-2815 OA AUDIT REPORT The report listed below was reviewed for compliance with the FDA Good Laboratory Practices and with EPA Good Laboratory Practices. The final report and all associated raw data were reviewed for accuracy and consistency and the findings were reported to management. The methods used were the methods described and the report accurately reflects the data. Therefore, these studies were done in compliance with the FDA and EPA Good Laboratory Practices. Franklin B. Newman QA Auditor SPONSOR: flfifcVS PiS STUDY: tjftotf REPORT TYPE: CfiOL AUDIT DATE: REPORT TO MANAGEMENT: 9 -1 5 -0 ) AUDIT #: 060574 418-018:PAGE H-78 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No. 1028 Study: 418018 F0 C/S GD21 Species: RAT Accession Sgec. No. B 0037155 10901 B 0037156 10902 B 0037157 10903 B 0037158 10904 B 0037159 10905 B 0037160 10907 B 0037161 10908 B 0037162 10910 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F LDL-DIR MG/GM 0.63 0.85 0.53 0.00 0.65 0.34 0.49 0.80 .536 .272 HDL-DIR MG/GM 0.56 0.65 0.50 0.28 0.54 0.60 0.58 0.63 .543 .116 CHOLEST MG/GM 2.83 3.49 2.93 2.43 2.65 3.50 3.07 3.16 3.008 .378 TRIG MG/GM 7.97 7.44 7.09 6.03 6.76 11.47 8.72 8.27 7.969 1.656 B 0037211 11040 B 0037212 11044 B 0037213 11045 B 0037214 11046 B 0037215 11047 B 0037216 11050 B 0037217 11051 B 0037218 11052 Mean S.D. Group 12 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 1.03 0.78 0.35 0.68 1.09 0.67 0.55 0.69 .73 .241 0.68 0.58 0.37 0.58 0.68 0.56 0.49 0.54 .56 .101 2.38 2.45 2.94 2.66 2.87 2.49 2.77 2.44 2.625 .215 6.99 8.63 7.26 8.38 8.59 7.38 6.90 7.62 7.719 .713 Reference Range 0- 0- 00 Page 1 + 0- 0 0- 0 000575 418-018:PAGE H-79 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client ARGUS RESEARCH LABORATORIES, INC 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 F0 C/S GD21 Species : RAT Accession Spec. No. ID B 0037219 11053 B 0037220 11054 B 0037221 11056 B 0037222 11058 B 0037223 11060 B 0037224 11062 B 0037225 11063 B 0037226 11066 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F LDL-DIR MG/GM 0.73 0.78 0.70 0.58 0.45 0.66 0.45 0.56 .614 .124 HDL-DIR MG/GM 0.58 0.60 0.52 0.48 0.40 0.52 0.50 0.48 .51 .062 CHOLEST MG/GM 2.68 2.68 2.13 2.68 1.99 2.37 2.45 2.36 2.418 .262 TRIG MG/GM 6.83 7.87 7.23 6.70 5.15 6.55 5.83 6.67 6.604 .826 B 0037163 10915 B 0037164 10916 B 0037165 10917 B 0037166 10918 B 0037167 10919 B 0037168 10920 B 0037169 10925 B 0037170 10928 Mean S.D. Group 2 2F 2F 2F 2F 2F 2F 2F 2F 0.80 0.55 0.38 0.00 0.60 0.55 0.47 0.28 .454 .24 0.59 0.52 0.53 0.35 0.55 0.59 0.49 0.50 .515 .076 3.01 2.81 3.65 2.58 3.33 3.15 2.81 2.89 3.029 .34 7.98 8.59 11.86 8.23 9.44 10.33 10.37 11.36 9.77 1.447 Reference Range 00 00 Page 2 + 0 0 00 00057 418-018:PAGE H-80 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client ARGUS RESEARCH LABORATORIES, INC 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 F0 C/S GD21 Species: RAT Accession Spec. No. ID B 0037171 10930 B 0037172 10931 B 0037173 10932 B 0037174 10933 B 0037175 10934 B 0037176 10935 B 0037177 10937 B 0037178 10940 Mean S.D. Group 3 Gp Sex Age 3F 3F 3F 3F 3F 3F 3F 3F LDL-DIR MG /GM 0.80 0.92 0.82 0.80 0.83 0.96 0.87 0.71 .839 .078 HDL-DIR MG/GM 0.59 0.63 0.63 0.56 0.65 0.63 0.60 0.56 .606 .034 CHOLEST MG/GM 2.58 2.62 2.54 2.43 2.29 2.68 2.60 2.51 2.531 .123 TRIG MG /GM 7.02 7.90 8.33 6.65 7.15 7.52 7.66 7.89 7.515 .548 B 0037179 10945 B 0037180 10946 B 0037181 10947 B 0037182 10949 B 0037183 10950 B 0037184 10951 B 0037185 10953 B 0037186 10954 Mean S.D. Group 4 4F 4F 4F 4F 4F 4F 4F 4F 0.71 0.68 0.75 0.71 0.75 1.04 0.69 0.73 .758 .117 0.62 0.56 0.55 0.54 0.52 0.68 0.53 0.56 .57 .054 3.01 2.10 2.30 2.51 2.06 2.28 2.43 2.22 2.364 .302 9.91 -6.67 6.69 5.62 6.17 7.98 6.91 6.30 7.031 1.348 Reference Range 0- o- 00 Page 3 + 0- 0 0- 0 000577 418-018:PAGE H-81 X L X Z -^ jL JL X v A y INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 F0 C/S GD21 Species: RAT Accession Spec. No. ID B 0037187 10958 B 0037188 10959 B 0037189 10960 B 0037190 10961 B 0037191 10962 B 0037192 10963 B 0037193 10964 B 0037194 10965 Mean S.D. Group 5 Gp Sex Age 5F 5F 5F 5F 5F 5F 5F 5F LDL-DIR MG/GM 0.66 0.12 0.01 0.51 0.25 0.00 0.00 0.61 .27 .283 HDL-DIR MG/GM 0.66 0.57 0.43 0.62 0.47 0.34 0.38 0.65 .515 .126 CHOLEST MG/GM 3.07 3.32 3.11 2.90 2.70 3.22 2.41 3.12 2.981 .299 TRIG MG /GM 10.90 10.77 9.81 8.73 8.43 8.46 10.17 9.23 9.563 .998 B 0037195 10972 B 0037196 10974 B 0037197 10975 B 0037198 10976 B 0037199 10978 B 0037200 10980 B 0037201 10981 B 0037202 10982 Mean S.D. Group 6 6F 6F 6F 6F 6F 6F 6F 6F 0.71 0.49 0.97 1.07 0.46 0.92 0.93 0.90 .806 .228 0.69 0.57 0.64 0.74 0.42 0.65 0.69 0.61 .626 .098 2.90 2.88 2.19 3.13 2.66 2.56 2.90 2.56 2.723 .291 8.51 7.47 6.32 8.27 7.59 7.88 8.42 7.35 7.726 .719 Reference Range 0- 0- 0- 00000 Page 4 + 0G0578 418-018:PAGE H-82 JL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 F0 C/S GD21 Species: RAT Accession Spec. No. ID B 0037203 10985 B 0037204 10987 B 0037205 10988 B 0037206 10989 B 0037207 10990 B 0037208 10991 B 0037209 10992 B 0037210 10993 Mean S.D. Group 7 Gp Sex Age 7F 7F 7F 7F 7F 7F 7F 7F LDL-DIR MG/GM 0.46 0.78 0.81 1.03 0.65 0.75 0.92 0.92 .79 .178 HDL-DIR MG/GM 0.45 0.59 0.63 0.69 0.51 0.57 0.69 0.69 .603 .09 CHOLEST MG/GM 2.75 2.39 2.74 2.47 2.83 3.01 2.75 2.52 2.683 .207 TRIG MG/GM 6.58 7.69 7.48 7.30 7.45 7.82 8.98 7.00 7.538 .703 Reference Range 0- 0- 0- 00 000 Approved -- Page 5 of 5 Date 000579 418-018:PAGE H-83 Client: INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No,. 1028 Study: 418018 F0 LD5 REPI Species: RAT Accession Sgec. No. B 0036757 10906 B 0036758 10909 B 0036759 10911 B 0036760 10912 B 0036761 10913 B 0036762 10914 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F LDL-DIR MG/GM 0.00 0.00 0.06 0.00 0.00 0.00 .01 .024 HDL-DIR MG/GM 0.17 0.38 0.31 0.18 0.26 0.24 .257 .08 CHOLEST MG/GM 3.26 5.56 3.06 3.97 3.72 5.60 4.195 1.12 TRIG MG/GM 9.43 12.82 10.39 14.78 8.18 16.39 11.998 3.208 B 0036813 11019 B 0036814 11021 B 0036815 11023 B 0036816 11024 B 0036817 11025 B 0036818 11026 B 0036819 11027 B 0036820 11028 B 0036839 11020 Mean S.D. Group 10 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 0.00 0.00 0.00 0.00 0.02 0.00 0.00 0.00 0.00 .002 .007 0.18 0.33 0.21 0.22 0.37 0.31 0.36 0.24 0.34 .284 .072 2.85 2.78 2.90 2.85 3.00 2.88 2.99 2.57 3.35 2.908 .209 9.53 8.38 7.51 7.94 11.05 9.13 10.12 7.56 9.96 9.02 1.249 Reference Range 0- 0- 0- 0 0 000 Page 1 + 000580 418-018:PAGE H-84 ^4 T JL JL V ^ K / 200 Girard Street, Suite 200, Gaithersburg, MD 20877 INCORPORATED 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No . 1028 Study: 418018 F0 LD5 REPI Species: RAT Accession Sgec. No. B 0036821 11029 B 0036822 11031 B 0036823 11032 B 0036824 11033 B 0036825 11034 B 0036826 11036 B 0036827 11037 B 0036828 11038 B 0036840 11035 Mean S.D. Group 11 Gp Sex Age 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F LDL-DIR MG/GM 0.15 0.31 0.33 0.01 0.00 0.00 0.00 0.08 0.10 .109 .131 HDL-DIR MG/GM 0.41 0.49 0.48 0.40 0.31 0.28 0.28 0.37 0.35 .374 .078 CHOLEST MG/GM 2.73 3.10 4.24 3.55 3.91 2.76 2.77 2.91 3.37 3.26 .548 TRIG MG/GM 8.47 10.25 19.83 14.02 15.46 9.13 7.62 7.89 9.96 11.403 4.156 B 0036829 11041 B 0036830 11049 B 0036831 11042 B 0036832 11048 Mean S.D. Group 12 12 F 12 F 12 F 12 F 0.00 0.15 0.00 0.00 .038 .075 0.42 0.44 0.25 0.32 .358 .089 3.39 3.39 3.45 4.26 3.623 .426 13.06 14.85 16.06 15.60 14.893 1.32 B 0036833 11059 Reference Range 13 F 0.19 00 0.45 00 3.26 00 9.58 00 Page 2 + 0005! 418-018:PAGE H-85 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No., 1028 Study: 418018 F0 LD5 REPI Species: RAT Accession Sgec. No. B 0036834 11055 B 0036835 11057 B 0036836 11061 B 0036837 11064 B 0036838 11065 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F 13 F 13 F LDL-DIR MG/GM 0.00 0.00 0.00 0.13 0.00 .053 .085 HDL-DIR MG/GM 0.33 0.26 0.22 0.47 0.45 .363 .108 CHOLEST MG/GM 6.52 3.50 4.11 3.38 3.79 4.093 1.228 TRIG MG/GM 22.98 15.80 19.26 11.96 1 .61 15.865 4.837 B 0036763 10921 B 0036764 10922 B 0036765 10923 B 0036766 10924 B 0036767 10927 Mean S.D. Group 2 2F 2F 2F 2F 2F 0.11 0.00 0.14 0.00 0.00 .05 .069 0.37 0.27 0.51 0.13 0.29 .314 .14 3.59 4.31 4.13 2.91 3.84 3.756 .547 9.14 8.94 10.76 11.73 10.75 10.264 1.188 B 0036768 10929 B 0036769 10938 B 0036770 10941 B 0036771 10942 Mean S.D. Group 3 3F 3F 3F 3F 0.23 0.00 0.00 0.16 .098 .116 0.53 0.29 0.41 0.44 .418 .099 3.34 2.78 3.91 3.07 3.275 .481 8.09 6.59 10.62 8.54 8.46 1.664 Reference Range 0- 000 Page 3 + 0 0 0 0 0C0582 418-018:PAGE H-86 o^zNmmcs. INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No,. 1028 study: 418018 F0 LD5 REPI Species: RAT Accession Spec. No. ID B 0036772 10943 B 0036773 10944 B 0036774 10948 B 0036775 10952 B 0036776 10955 B 0036777 10956 Mean S.D. Group 4 Gp Sex Age 4F 4F 4F 4F 4F 4F LDL-DIR MG/GM 0.00 0.00 0.00 0.13 0.00 0.00 .022 .053 HDL-DIR MG/GM 0.26 0.37 0.42 0.48 0.23 0.31 .345 .096 CHOLEST MG/GM 2.95 b 5b 3.51 5.52 2.89 3.27 4.117 1.542 TRIG MG/GM 18.57 14.80 15.15 13.06 9.21 10.87 13.61 3.333 B 0036778 10957 B 0036779 10966 B 0036780 10967 B 0036781 10968 B 0036782 10969 B 0036783 10970 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 0.00 0.02 0.00 0.00 0.00 0.00 .003 .008 0.30 0.35 0.21 0.27 0.28 0.27 .28 .046 3.08 2.89 2.95 3.14 3.35 2.67 3.013 .233 9.80 7.95 10.65 7.51 8.80 9.03 8.957 1.159 B 0036784 10971 B 0036785 10973 Reference Range 6F 6F 0.00 0.00 00 0.28 0.27 00 4.16 3.42 00 11.02 18.88 00 Page 4 + 000583 418-018-.PAGE H-87 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No,. 1028 Study: 418018 F0 LD5 REPI Species: RAT Accession Sgec. No. B 0036786 10977 B 0036787 10983 B 0036788 10979 B 0036789 10984 Mean S.D. Group 6 Gp Sex Age 6F 6F 6F 6F LDL-DIR MG/GM 0.00 0.01 0.16 0.00 .028 .065 HDL-DIR MG/GM 0.42 0.44 0.45 0.23 .348 .099 CHOLEST MG/GM 2.81 3.13 2.90 2.83 3.208 .52 TRIG MG/GM 9.54 8.10 9.61 13.41 11.76 3.922 B 0036790 10995 B 0036791 10997 B 0036792 10986 B 0036793 10994 B 0036794 10996 Mean S.D. Group 7 7F 7F 7F 7F 7F 0.00 0.00 0.00 0.00 0.00 0 0 0.37 0.26 0.29 0.36 0.22 .3 .064 3.46 2.72 3.76 3.25 2.72 3.182 .459 10.85 6.25 12.58 18.69 9.35 11.544 4.622 B 0036795 10999 B 0036796 11000 B 0036797 11001 B 0036798 11002 B 0036799 11003 Reference Range 8F 8F 8F 8F 8F 0.00 0.00 0.00 0.00 0.00 00 0.32 0.31 0.34 0.35 0.32 00 3.04 3.34 3.23 3.02 3.00 00 8.57 7.55 10.31 8.00 9.66 00 Page 5 + 0005*4 418-018:PAGE H-88 V * j I Ji Jl v A - / INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No,. 1028 Study: 418018 F0 LD5 REPI Species: RAT Accession Spec. No. ID B 0036800 11004 B 0036801 11005 B 0036802 11006 B 0036803 11007 B 0036804 11008 Mean S.D. Group 8 Gp Sex Age 8F 8F 8F 8F 8F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.15 .015 .047 HDL-DIR MG/GM 0.27 0.34 0.25 0.23 0.47 .32 .066 CHOLEST MG/GM 3.21 3.08 3.21 3.35 3.45 3.193 .155 TRIG MG/GM 10.48 11.49 7.95 7.79 8.06 8.986 1.386 B 0036805 11009 B 0036806 11010 B 0036807 11011 B 0036808 11012 B 0036809 11013 B 0036810 11015 B 0036811 11016 B 0036812 11018 Mean S.D. Group 9 9F 9F 9F 9F 9F 9F 9F 9F 0.00 0.22 0.20 0.00 0.13 0.27 0.00 0.00 .103 .116 0.29 0.39 0.46 0.33 0.40 0.43 0.19 0.27 .345 .091 2.86 2.96 3.38 2.99 2.90 3.07 2.95 2.86 2.996 .17 6.53 7.32 9.65 7.32 7.98 8.30 9.19 7.82 8.014 1.024 Reference Range 0nu 0- 0- au y. un A p p r o v e d --- ---------------Page & of 6 0- AU Date 000585 418-018:PAGE H-89 ^LA jL^LL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MO 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No,. 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Spec. No. ID B 0036600 11501 B 0036601 11502 B 0036602 11503 B 0036603 11504 B 0036604 11505 B 0036605 11506 B 0036606 11507 B 0036607 11508 B 0036608 11509 B 0036609 11510 B 0036610 11511 B 0036611 11512 B 0036612 11513 B 0036613 11514 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.09 0.11 0.11 0.07 0.08 0.08 0.05 0.09 0.06 0.08 0.09 0.08 0.09 0.10 .084 .017 CHOLEST MG/GM 1.43 1.66 1.79 1.76 1.81 1.75 1.63 1.57 1.68 1.71 1.75 1.60 1.93 1.95 1.716 .138 TRIG MG/GM 5.45 6.19 5.49 6.72 6.36 5.79 8.69 5.08 5.85 7.33 6.38 6.07 8.51 5.74 6.404 1.088 B 0036712 11620 B 0036713 11621 B 0036714 11622 B 0036715 11623 B 0036716 11624 B 0036717 11625 B 0036718 11626 Reference Range 10 F 10 F 10 F 10 F 10 F 10 F 10 F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 o0 0.14 0.10 0.14 0.17 0.09 0.11 0.12 00 1.89 1.99 2.46 2.04 1.78 1.90 2.11 00 8.20 6.76 22.27 6.65 7.93 6.88 6.29 00 Page 1 + 000586 418-018:PAGE H-90 JL JL v A J INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MO 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No.. 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Sgec. No. B 0036719 11627 B 0036720 11628 B 0036721 11629 Mean S.D. Group 10 Gp Sex Age 10 F 10 F 10 F LDL-DIR MG/GM 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.16 0.15 0.15 .133 .027 CHOLEST MG/GM 3.30 1.83 1.86 2.116 .459 TRIG MG/GM 59.25 7.40 10.48 14.211 16.528 B 0036722 11630 B 0036723 11632 B 0036724 11633 B 0036725 11634 B 0036726 11635 B 0036727 11636 B 0036728 11637 B 0036729 11638 B 0036730 11639 Mean S.D. Group 11 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.14 0.11 0.18 0.09 0.14 0.17 0.13 0.12 0.14 .136 .028 1.79 1.73 1.78 1.78 1.78 1.91 1.81 1.78 2.05 1.823 .098 8.04 9.16 7.66 7.91 9.92 7.01 7.81 5.88 9.00 8.043 1.207 B 0036731 11640 B 0036732 11641 Reference Range 12 F 12 F 0.00 0.00 00 0.13 0.12 00 1.69 1.76 00 8.68 12.72 00 Page 2 + 000587 418-018:PAGE H-91 XLLJL^dl. JL I V w * 200 Girard Street, Suite 200, Gaithersburg, MD 20877 INCORPORATED 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Sgec. No. B 0036733 11642 B 0036734 11643 B 0036735 11644 B 0036736 11645 B 0036737 11646 B 0036738 11647 B 0036739 11648 B 0036740 11649 B 0036741 11650 B 0036742 11651 B 0036743 11653 Mean S.D. Group 12 Gp Sex Age 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.05 0.00 0.00 0.00 0.00 0.00 .004 .014 HDL-DIR MG/GM 0.10 0.16 0.13 0.20 0.17 0.29 0.15 0.28 0.15 0.15 0.16 .168 .057 CHOLEST MG/GM 1.92 2.10 1.83 2.27 2.20 3.59 2.22 3.14 2.20 1.88 2.36 2.243 .548 TRIG MG/GM 8.11 8.94 7.96 20.75 10.80 29.18 7.64 29.88 7.64 8.43 6.95 12.898 8.221 B 0036744 11654 B 0036745 11655 B 0036746 11656 B 0036747 11657 B 0036748 11658 B 0036749 11659 B 0036750 11660 B 0036751 11661 B 0036752 11662 B 0036753 11663 Reference Range 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 13 F 0.03 0.00 0.00 0.00 0.08 0.00 0.00 0.00 0.00 0.00 00 0.11 0.15 0.11 0.18 0.18 0.15 0.16 0.16 0.14 0.18 00 2.26 1.94 1.89 2.98 2.64 2.72 2.61 2.09 2.09 2.65 00 10.47 10.79 8.29 19.04 13.78 19.86 19.46 8.88 28.49 17.65 00 Page 3 + 000588 418-018:PAGE H-92 JL X v A J INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No,. 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Spec. No. ID B 0036754 11664 B 0036755 11665 B 0036756 11667 Mean S.D. Group 13 Gp Sex Age 13 F 13 F 13 F LDL-DIR MG/GM 0.00 0.08 0.00 .015 .03 HDL-DIR MG/GM 0.11 0.23 0.19 .158 .035 CHOLEST MG/GM 2.01 2.59 2.42 2.376 .35 TRIG MG/GM 16.44 21.77 14.12 16.08 5.806 B 0036614 11515 B 0036615 11516 B 0036616 11517 B 0036617 11518 B 0036618 11519 B 0036619 11520 B 0036620 11521 B 0036621 11523 B 0036622 11524 B 0036623 11525 B 0036624 11526 B 0036625 11527 B 0036626 11528 Mean S.D. Group 2 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.12 0.09 0.07 0.08 0.12 0.11 0.12 0.10 0.11 0.09 0.08 0.08 0.09 .097 .018 1.76 1.73 1.31 1.91 1.77 1.56 2.13 1.57 1.49 1.92 1.80 1.63 1.66 1.711 .211 6.38 5.08 6.11 6.59 6.06 6.35 9.30 6.28 6.35 6.89 5.56 7.44 5.48 6.452 1.051 Reference Range 0- 0- 0- 00 0 00 Page 4 + 0C0589 418-018:PAGE H-93 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 F0 LD5 REP2 Species : RAT Accession Spec. No. ID B 0036627 11529 B 0036628 11530 B 0036629 11532 B 0036630 11533 B 0036631 11534 B 0036632 11535 B 0036633 11536 B 0036634 11537 B 0036635 11538 B 0036636 11539 B 0036637 11540 B 0036638 11541 B 0036639 11542 B 0036640 11543 Mean S.D. Group 3 B 0036641 11544 B 0036642 11545 B 0036643 11546 B 0036644 11547 B 0036645 11549 B 0036646 11550 B 0036647 11551 Reference Range Gp Sex Age 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.00 0.00 0.01 0.00 .002 .006 HDL-DIR MG/GM 0.16 0.14 0.13 0.12 0.10 0.13 0.11 0.13 0.15 0.11 0.12 0.12 0.06 0.15 .124 .025 CHOLEST MG/GM 1.79 1.75 2.09 2.23 1.81 1.97 1.97 1.60 2.03 2.10 2.38 2.39 1.41 1.86 1.956 .279 TRIG MG /GM 5.60 6.28 7.69 10.26 10.89 8.99 9.04 6.01 8.44 10.65 28.91 24.63 8.05 6.86 10.879 6.987 4F 4F 4F 4F 4F 4F 4F 0.00 0.04 0.00 0.01 0.00 0.00 0.00 00 0.08 0.09 0.04 0.14 0.08 0.15 0.16 00 1.84 2.17 2.02 2.29 2.09 2.35 2.03 00 16.82 30.38 16.20 18.27 21.34 12.24 10.45 00 Page 5 + 000590 418-018:PAGE H-94 SJLJLm JJL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Spec. No. ID B 0036648 11552 B 0036649 11553 B 0036650 11554 B 0036651 11555 B 0036652 11556 B 0036653 11557 Mean S.D. Group 4 Gp Sex Age 4F 4F 4F 4F 4F 4F LDL-DIR MG/GM 0.03 0.00 0.00 0.00 0.00 0.00 .006 .013 HDL-DIR MG/GM 0.14 0.11 0.06 0.12 0.12 0.08 .105 .037 CHOLEST MG/GM 2.05 2.05 1.77 1.92 2.02 1.92 2.04 .164 TRIG MG/GM 10.22 10.47 12.68 12.01 24.42 11.83 15.948 6.227 B 0036654 11558 B 0036655 11559 B 0036656 11561 B 0036657 11563 B 0036658 11564 B 0036659 11565 B 0036660 11567 B 0036661 11568 B 0036662 11569 B 0036663 11570 B 0036664 11571 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 5F 5F 5F 5F 5F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.11 0.12 0.09 0.10 0.10 0.08 0.12 0.07 0.11 0.07 0.07 .095 .02 1.74 1.66 1.44 1.84 1.86 1.69 1.88 1.45 1.59 1.56 1.35 1.642 .181 6.38 5.20 6.22 9.49 7.20 5.80 9.37 6.94 6.64 5.55 5.96 6.795 1.428 Reference Range 0- 0 0- 0 Page 6 + 0 0 00 0C0591 418-018:PAGE H-95 X L X i - ^ J L JL x v A J INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Sgec. No. B 0036665 11572 B 0036666 11573 B 0036667 11574 B 0036668 11575 B 0036669 11576 B 0036670 11577 B 0036671 11578 B 0036672 11579 B 0036673 11580 B 0036674 11581 B 0036675 11582 B 0036676 11583 B 0036677 11584 B 0036678 11585 Mean S.D. Group 6 Gp Sex Age 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.14 0.11 0.16 0.14 0.12 0.11 0.15 0.16 0.12 0.13 0.08 0.15 0.13 0.18 .134 .026 CHOLEST MG /GM 2.47 1.92 1.95 1.69 1.88 2.08 1.95 2.19 1.77 1.80 1.80 1.67 1.67 2.02 1.919 .222 TRIG MG/GM 24.06 11.41 8.96 6.68 6.78 9.18 9.91 14.21 4.78 6.86 9.79 5.57 5.31 8.45 9.425 4.939 B 0036679 11586 B 0036680 11587 B 0036681 11588 B 0036682 11589 B 0036683 11590 B 0036684 11591 B 0036685 11592 Reference Range 7F 7F 7F 7F 7F 7F 7F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 00 0.08 0.14 0.14 0.18 0.14 0.13 0.18 00 2.86 2.05 2.07 2.24 1.96 1.88 1.88 00 23.94 14.70 8.33 18.17 7.11 7.80 13.17 00 Page 7 + 0C0592 418-018:PAGE H-96 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921-0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Spec. No. ID B 0036686 11593 B 0036687 11594 B 0036688 11595 B 0036689 11596 B 0036690 11597 B 0036691 11598 B 0036692 11599 Mean S.D. Group 7 Gp Sex Age 7F 7F 7F 7F 7F 7F 7F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.14 0.16 0.14 0.14 0.19 0.22 0.16 .153 .033 CH0LEST MG/GM 1.72 1.80 2.30 2.07 1.88 2.14 1.84 2.049 .287 TRIG MG/GM 8.00 8.52 8.05 9.62 9.26 7.88 5.99 10.753 5.057 B 0036693 11600 B 0036694 11601 B 0036695 11602 B 0036696 11603 B 0036697 11604 B 0036698 11605 B 0036699 11606 B 0036700 11607 B 0036701 11608 B 0036702 11609 Mean S.D. Group 8 8F 8F 8F 8F 8F 8F 8F 8F 8F 8F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.00 0.00 .002 .006 0.14 0.11 0.10 0.13 0.12 0.09 0.12 0.11 0.11 0.12 .115 .014 1.70 1.90 2.05 1.81 2.02 2.04 2.00 1.76 2.05 1.84 1.917 .132 4.77 6.46 7.21 8.74 9.98 10.15 7.80 9.19 5.41 8.55 7.826 1.846 Reference Range 0- 0 00 Page 8 + 0 0 00 000593 418-018:PAGE H-97 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 F0 LD5 REP2 Species: RAT Accession Spec. No. ID B 0036703 11611 B 0036704 11612 B 0036705 11613 B 0036706 11614 B 0036707 11615 B 0036708 11616 B 0036709 11617 B 0036710 11618 B 0036711 11619 Mean S.D. Group 9 Gp Sex Age 9F 9F 9F 9F 9F 9F 9F 9F 9F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.11 0.13 0.13 0.11 0.14 0.13 0.13 0.11 0.10 .121 .014 CHOLEST MG/GM 2.06 1.73 2.36 1.65 1.97 1.76 1.73 2.17 1.77 1.911 .243 TRIG MG/GM 11.98 6.11 16.42 7.18 8.63 6.32 9.22 9.19 7.77 9.202 3.243 Reference Range 0- 0- 0- 0- 00 0 0 Approved -- L_ Page 9 o f 9 Date -JJLjJZjjLL OC0594 418-018:PAGE H-98 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client ARGUS RESEARCH LABORATORIES, INC 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No. 1028 Study: 418018 Fl C/S GD21 Species: RAT Accession Sgec. No. B 0036958 10901 B 0036959 10902 B 0036960 10903 B 0036961 10904 B 0036962 10905 B 0036963 10907 B 0036964 10908 B 0036965 10910 Mean S.D. Group 1 B 0037014 11040 B 0037015 11044 B 0037016 11045 B 0037017 11046 B 0037018 11047 B 0037019 11050 B 0037020 11051 B 0037021 11052 Mean S.D. Group 12 Gp Sex Age 1 1 1 1 1 1 1 1 LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.02 0.00 0.01 0.01 0.02 0.00 0.02 0.00 .01 .009 CHOLEST MG/GM 3.26 3.76 2.89 3.88 3.71 3.59 3.10 2.99 3.398 .384 TRIG MG/GM 5.04 5.92 3.94 5.35 5.05 4.99 4.94 4.15 4.923 .63 12 0.00 0.02 3.28 5.06 12 0.00 0.02 3.21 5.13 12 12 12 0.00 0.00 0.00 0.01 0.03 0.03 3.70 3.18 4.32 6.04 5.95 7.90 12 0.00 0.02 3.26 5.18 12 12 0.00 0.00 0.02 0.02 3.32 3.36 6.16 5.48 0 .021 3.454 5.863 0 .006 .385 .931 OO 1 oo 1 1 oo Reference Range 00 Page 1 + 0G0595 418-018:PAGE H-99 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No. 1028 Study: 418018 Fl C/S GD21 Species: RAT Accession Spec. No. ID B 0037022 11053 B 0037023 11054 B 0037024 11056 B 0037025 11058 B 0037026 11060 B 0037027 11062 B 0037028 11063 B 0037029 11066 Mean S.D. Group 13 Gp Sex Age 13 13 13 13 13 13 13 13 LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.03 0.01 0.03 0.02 0.00 0.05 0.03 0.02 .024 .015 CHOLEST MG/GM 3.18 3.61 3.30 3.23 3.43 3.71 2.77 2.80 3.254 .341 TRIG MG/GM 5.82 5.99 5.21 6.37 5.91 6.92 4.27 4.48 5.621 .911 B 0036966 10915 B 0036967 10916 B 0036968 10917 B 0036969 10918 B 0036970 10919 B 0036971 10920 B 0036972 10925 B 0036973 10928 Mean S.D. Group 2 2 2 2 2 2 2 2 2 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.00 0.01 0.00 0.02 0.02 0.00 0.00 0.00 .006 .009 2.97 3.45 3.60 3.04 4.44 4.01 3.89 3.67 3.634 .491 4.79 5.04 4.96 4.42 8.40 4.99 5.60 5.24 5.43 1.247 Reference Range 00 o0 Page 2 + 0 0 00 000596 418-018:PAGE H -100 IN C O R P O R A T E D 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No . 1028 Study: 418018 FI C/S GD21 Species: RAT Accession Spec. No. ID B 0036974 10930 B 0036975 10931 B 0036976 10932 B 0036977 10933 B 0036978 10934 B 0036979 10935 B 0036980 10937 B 0036981 10940 Mean S.D. Group 3 Gp Sex Age 3 3 3 3 3 3 3 3 LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.03 0.05 0.07 0.03 0.05 0.04 0.01 0.02 .038 .019 CHOLEST MG/GM 5.09 4.88 4.69 4.31 5.10 4.10 4.32 3.81 4.538 .475 TRIG MG/GM 9.06 8.96 8.91 7.46 9.06 7.01 6.26 6.90 7.953 1.164 B 0036982 10945 B 0036983 10946 B 0036984 10947 B 0036985 10949 B 0036986 10950 B 0036987 10951 B 0036988 10953 B 0036989 10954 Mean S.D. Group 4 4 4 4 4 4 4 4 4 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.05 0.03 0.03 0.01 0.00 0.02 0.02 0.05 .026 .018 5.70 4.90 5.70 5.00 4.95 4.48 3.90 6.55 5.148 .82 10.53 9.96 10.06 9.71 8.50 7.39 6.25 14.22 9.578 2.387 Reference Range 0- 0- 0- 00000 Page 3 + 000597 418-018:PAGE H-101 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 INC. Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No.. 1028 Study: 418018 FI C/S GD21 Species : RAT Accession Spec. No. ID B 0036990 10958 B 0036991 10959 B 0036992 10960 B 0036993 10961 B 0036994 10962 B 0036995 10963 B 0036996 10964 B 0036997 10965 Mean S.D. Group 5 Gp Sex Age 5 5 5 5 5 5 5 5 LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.01 0.04 0.05 0.03 0.00 0.03 0.01 0.00 .021 .019 CHOLEST MG/GM 3.72 3.99 3.41 3.17 3.67 3.73 3.25 3.86 3.6 .293 TRIG MG/GM 5.39 7.02 5.45 4.75 5.04 5.77 4.80 5.72 5.493 .728 B 0036998 10972 B 0036999 10974 B 0037000 10975 B 0037001 10976 B 0037002 10978 B 0037003 10980 B 0037004 10981 B 0037005 10982 Mean S.D. Group 6 6 6 6 6 6 6 6 6 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.03 0.04 0.03 0.01 0.03 0.02 0.03 0.04 .029 .01 4.44 4.70 4.63 4.63 4.83 3.80 3.77 4.35 4.394 .404 7.99 9.46 7.79 7.92 9.08 5.99 6.11 7.94 7.785 1.23 l oo 1 oo 1 oo 1 oo Reference Range Page 4 + 000598 418-018:PAGE H -102 _ ^ /U \w < m cs INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/30/2001 Client No. 1028 Study: 418018 Fl C/S GD21 Species: RAT Accession Sgec. No. B 0037006 10985 B 0037007 10987 B 0037008 10988 B 0037009 10989 B 0037010 10990 B 0037011 10991 B 0037012 10992 B 0037013 10993 Mean S.D. Group 7 Gp Sex Age 7 7 7 7 7 7 7 7 LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.05 0.06 0.00 0.03 0.03 0.07 0.02 0.03 .036 .023 CHOLEST MG/GM 5.00 5.05 3.57 3.73 3.29 3.90 3.67 3.52 3.966 .677 TRIG MG/GM 8.88 8.29 8.04 6.39 6.18 7.07 6.09 5.88 7.103 1.154 Reference Range 00 0 0 0- 0 0- 0 Approved -- Page / Date 000599 418-018:PAGE H -103 JL^LL i . INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No., 1028 Study: 418018 Fl LD5 REPI Species: RAT Accession Sgec. No. B 0037092 10906 B 0037093 10909 B 0037094 10911 B 0037095 10913 B 0037096 10914 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.08 0.05 0.09 0.09 0.14 .09 .032 CHOLEST MG/GM 4.40 4.45 3.82 3.99 6.11 4.554 .91 TRIG MG/GM 29.86 21.02 11.37 8.79 26.27 19.462 9.169 B 0037030 10906 B 0037031 10909 B 0037032 10911 B 0037033 10913 B 0037034 10914 Mean S.D. Group 1 1M 1M 1M 1M 1M 0.00 0.00 0.00 0.00 0.00 0 0 0.06 0.06 0.10 0.07 0.11 .08 .023 3.94 3.83 4.00 3.51 4.82 4.02 .486 21.88 15.45 12.96 8.95 30.18 17.884 8.324 B 0037134 11019 B 0037135 11021 B 0037136 11023 B 0037137 11024 Reference Range 10 F 10 F 10 F 10 F 0.00 0.00 0.00 0.00 00 0.03 0.08 0.05 0.07 00 4.08 4.16 3.96 5.45 00 13.30 8.26 13.46 23.83 00 Page 1 + 000660 418-018:PAGE H-104 ^4 I i -Ji I V V J INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No,. 1028 Study: 418018 Fl LD5 REPI Species: RAT Accession Sgec. No. B 0037138 11025 B 0037139 11026 B 0037140 11027 B 0037141 11028 B 0037154 11020 Mean S.D. Group 10 Gp Sex Age 10 F 10 F 10 F 10 F 10 F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.11 0.10 0.05 0.04 0.10 .07 .029 CHOLEST MG/GM 5.52 4.05 4.66 3.39 4.35 4.402 .7 TRIG MG /GM 16.93 9.43 12.57 10.10 10.29 13.13 4.796 B 0037072 11019 B 0037073 11021 B 0037074 11023 B 0037075 11024 B 0037076 11025 B 0037077 11026 B 0037078 11027 B 0037079 11028 B 0037091 11020 Mean S.D. Group 10 10 M 10 M 10 M 10 M 10 M 10 M 10 M 10 M 10 M 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.08 0.11 0.09 0.07 0.14 0.05 0.11 0.14 0.07 .096 .032 4.22 4.42 4.12 4.16 4.38 3.99 3.89 3.80 5.30 4.253 .444 12.97 8.95 13.31 16.23 13.38 9.97 8.79 9.09 9.74 11.381 2.653 Reference Range 0- 0- 0- 00000 Page 2 + 0008.01 418-018:PAGE H-105 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REPI Species: RAT Accession Spec. No. ID B 0037142 11029 B 0037143 11031 B 0037144 11032 B 0037145 11033 B 0037146 11034 B 0037147 11035 B 0037148 11036 B 0037149 11037 B 0037150 11038 Mean S.D. Group 11 Gp Sex Age 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.04 0.07 0.05 0.08 0.14 0.06 0.06 0.08 0.03 .068 .032 CHOLEST MG/GM 3.78 4.26 4.14 3.79 4.04 3.87 4.02 3.98 3.49 3.93 .228 TRIG MG/GM 9.56 19.52 17.52 10.94 5.10 6.53 9.08 13.43 8.40 11.12 4.85 B 0037080 11029 B 0037081 11031 B 0037082 11032 B 0037083 11033 B 0037084 11034 B 0037085 11035 B 0037086 11036 B 0037087 11037 B 0037088 11038 Mean S.D. Group 11 11 M 11 M 11 M 11 M 11 M 11 M 11 M 11 M 11 M 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.11 0.08 0.05 0.09 0.06 0.08 0.07 0.03 0.03 .067 .027 3.44 4.15 4.27 4.14 4.43 4.31 4.14 4.06 3.25 4.021 .402 6.71 9.86 20.08 8.17 10.71 7.52 8.99 11.59 9.21 10.316 3.964 Reference Range 00 00 Page 3 + 0 0 0o 0CGt 02 418-018:PAGE H-106 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REP1 Species: RAT Accession Spec. No. ID B 0037151 11041 B 0037152 11049 Mean S.D. Group 12 Gp Sex Age 12 F 12 F LDL-DIR MG/GM 0.00 0.00 0 0 HDL-DIR MG/GM 0.09 0.13 .11 .028 CHOLEST MG/GM 4.13 3.43 3.78 .495 TRIG MG/GM 12.91 12.04 12.475 .615 B 0037089 11049 Mean S.D. Group 12 12 M 0.00 0 0 0.07 .07 0 3.97 3.97 0 8.82 8.82 0 B 0037153 11059 Mean S.D. Group 13 13 F 0.00 0 0 0.07 .07 0 4.39 4.39 0 11.96 11.96 0 B 0037090 11059 Mean S.D. Group 13 13 M 0.00 0 0 0.04 .04 0 3.31 3.31 0 8.45 8.45 0 Reference Range 00 00 Page 4 + 0 0 00 003 418-018:PAGE H -107 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No., 1028 Study: 418018 Fl LD5 REPI Species : RAT Accession Sgec. No. B 0037097 10921 B 0037098 10922 B 0037099 10923 B 0037100 10924 B 0037101 10927 Mean S.D. Group 2 Gp Sex Age 2F 2F 2F 2F 2F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.05 0.14 0.04 0.10 0.05 .076 .043 CHOLEST MG/GM 4.26 4.63 3.17 4.73 4.00 4.158 .625 TRIG MG/GM 22.52 30.62 16.50 9.40 32.85 22.378 9.754 B 0037035 10921 B 0037036 10922 B 0037037 10923 B 0037038 10924 B 0037039 10927 Mean S.D. Group 2 2M 2M 2M 2M 2M 0.00 0.00 0.00 0.00 0.00 0 0 0.05 0.12 0.08 0.11 0.07 .086 .029 4.40 4.75 4.44 4.42 3.99 4.4 .27 12.79 23.41 14.33 7.43 35.20 18.632 10.902 B 0037102 10929 B 0037103 10938 B 0037104 10942 Mean S.D. Group 3 3F 3F 3F 0.00 0.00 0.00 0 0 0.09 0.08 0.05 .073 .021 5.16 3.98 3.72 4.287 .767 27.88 15.75 15.74 19.79 7.006 Reference Range 00 00 Page 5 + 0 0 00 OOOG04 418-018:PAGE H -108 J-^ LL JL i . V A J INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No,. 1028 Study: 418018 Fl LD5 REPI Species: RAT Accession Sgec. No. B 0037040 10929 B 0037041 10938 B 0037042 10942 Mean S.D. Group 3 Gp Sex Age 3M 3M 3M LDL-DIR MG/GM 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.06 0.04 0.04 .047 .012 CHOLEST MG/GM 5.03 3.65 3.83 4.17 .75 TRIG MG /GM 24.76 9.29 19.75 17.933 7.893 B 0037105 10957 B 0037106 10966 B 0037107 10967 B 0037108 10968 B 0037109 10969 B 0037110 10970 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.04 0.09 0.07 0.03 0.09 0.06 .063 .025 3.95 4.04 4.13 3.82 4.17 3.96 4.012 .129 16.66 20.88 14.94 14.56 27.74 21.63 19.402 5.051 B 0037043 10957 B 0037044 10966 B 0037045 10967 B 0037046 10968 B 0037047 10969 Reference Range 5M 5M 5M 5M 5M 0.00 0.00 0.00 0.00 0.00 00 0.04 0.03 0.11 0.07 0.05 00 4.51 3.22 3.66 3.93 4.07 00 19.77 26.21 19.79 17.13 18.42 00 Page 6 + 0C0CO5 418-018:PAGE H-109 Jf V I INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MO 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REP1 Species: RAT Accession Spec. No. ID B 0037048 10970 Mean S.D. Group 5 Gp Sex Age 5M LDL-DIR MG/GM 0.00 0 0 HDL-DIR MG/GM 0.08 .063 .029 CHOLEST MG/GM 3.92 3.885 .429 TRIG MG/GM 27.48 21.467 4.3 B 0037111 10977 B 0037112 10979 B 0037113 10983 Mean S.D. Group 6 6F 6F 6F 0.00 0.00 0.00 0 0 0.07 0.05 0.02 .047 .025 3.66 4.14 4.38 4.06 .367 12.43 17.90 16.15 15.493 2.793 B 0037049 10977 B 0037050 10979 B 0037051 10983 Mean S.D. Group 6 6M 6M 6M 0.00 0.00 0.00 0 0 0.07 0.12 0.05 .08 .036 3.80 3.96 3.62 3.793 .17 17.37 9.84 11.76 12.99 3.913 Reference Range 0- 0- 0- 00000 Page 7 + oooens 418-018:PAGE H-110 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 Fl LD5 REPI Species : RAT Accession Sgec. No. B 0037114 10995 B 0037115 10997 Mean S.D. Group 7 Gp Sex Age 7F 7F LDL-DIR MG/GM 0.00 0.00 0 0 HDL-DIR MG/GM 0.14 0.05 .095 .064 CHOLEST MG /GM 3.81 4.02 3.915 .148 TRIG MG/GM 7.37 11.63 9.5 3.012 B 0037052 10995 B 0037053 10997 Mean S.D. Group 7 7M 7M 0.00 0.00 0 0 0.04 0.08 .06 .028 4.55 4.49 4.52 .042 15.32 7.92 11.62 5.233 B 0037116 10999 B 0037117 11000 B 0037118 11001 B 0037119 11002 B 0037120 11003 B 0037121 11004 B 0037122 11005 B 0037123 11006 B 0037124 11007 B 0037125 11008 Mean S.D. Group 8 8F 8F 8F 8F 8F 8F 8F 8F 8F 8F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.07 0.02 0.09 0.06 0.07 0.09 0.07 0.06 0.08 0.05 .066 .021 3.45 3.89 3.66 4.40 4.40 4.24 4.02 4.17 4.07 3.62 3.992 .331 12.00 24.76 5.34 11.96 16.38 9.45 16.78 9.86 14.07 8.73 12.933 5.442 Reference Range 0- 0- 0- 00000 Page 8 + 000607 418-018:PAGE H - l l l INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported:. 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REPI Species : RAT Accession Spec. No. ID B 0037054 10999 B 0037055 11000 B 0037056 11001 B 0037057 11002 B 0037058 11003 B 0037059 11004 B 0037060 11005 B 0037061 11006 B 0037062 11007 B 0037063 11008 Mean S.D. Group 8 Gp Sex Age 8M 8M 8M 8M 8M 8M 8M 8M 8M 8M LDL-DIR MG /GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.04 0.08 0.08 0.07 0.11 0.05 0.06 0.00 0.22 0.07 .078 .058 CHOLEST MG/GM 3.90 4.65 3.87 4.05 4.47 3.44 3.85 4.26 6.47 4.98 4.394 .855 TRIG MG/GM 16.84 28.77 7.18 13.48 29.31 7.92 15.88 13.43 20.61 19.78 17.32 7.565 B 0037126 11009 B 0037127 11010 B 0037128 11011 B 0037129 11012 B 0037130 11013 B 0037131 11015 B 0037132 11016 B 0037133 11018 Mean S.D. Group 9 9F 9F 9F 9F 9F 9F 9F 9F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.04 0.03 0.09 0.07 0.04 0.06 0.08 0.09 .063 .024 4.15 3.86 3.97 4.21 3.46 4.20 4.40 3.66 3.989 .315 10.70 14.04 8.39 10.84 10.94 7.80 17.91 16.75 12.171 3.708 Reference Range 00 00 Page 9 + 0 0 00 000608 418-018:PAGE H -112 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/07/2001 Date Reported: 10/10/2001 Client No., 1028 Study: 418018 FI LD5 KEPI Species: RAT Accession Spec. No. ID B 0037064 11009 B 0037065 11010 B 0037066 11011 B 0037067 11012 B 0037068 11013 B 0037069 11015 B 0037070 11016 B 0037071 11018 Mean S.D. Group 9 Gp Sex Age 9M 9M 9M 9M 9M 9M 9M 9M LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.03 0.02 0.02 0.06 0.07 0.10 0.07 0.07 .055 .029 CHOLEST MG/GM 4.14 3.70 4.06 4.06 4.03 3.10 4.63 4.42 4.018 .463 TRIG MG/GM 13.73 11.29 11.14 7.78 14.71 5.07 17.71 23.50 13.116 5.761 Reference Range 00 00 0 0 00 Approved -- Page Date .JsJlsJ&L 000603 418-018:PAGE H-113 1 f v i 1 JL v A y INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No,. 1028 Study: 418018 Fl LD5 REP2 Species : RAT Accession Spec. No. ID B 0037431 11501 B 0037432 11502 B 0037433 11503 B 0037434 11504 B 0037435 11505 B 0037436 11506 B 0037437 11507 B 0037438 11508 B 0037439 11509 B 0037440 11510 B 0037441 11511 B 0037442 11512 B 0037443 11513 B 0037444 11514 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F 1F iF 1F 1F 1F 1F LDL-DIR MG/GM 0.00 0.05 0.00 0.00 0.03 0.00 0.00 0.00 0.00 0.03 0.10 0.00 0.00 .016 .03 HDL-DIR MG/GM 0.10 0.09 0.13 0.12 0.16 0.08 0.09 0.11 0.11 0.10 0.10 0.09 0.09 .105 .021 CHOLEST MG/GM 3.51 3.19 3.54 3.50 3.29 3.37 3.86 3.70 3.73 3.46 3.71 3.67 3.91 3.572 .215 TRIG MG/GM 18.40 7.39 29.23 22.39 10.50 18.51 32.22 24.47 17.71 10.33 9.15 14.90 13.41 17.585 7.769 B 0037325 11501 B 0037326 11502 B 0037327 11503 B 0037328 11504 B 0037329 11505 B 0037330 11506 B 0037331 11507 Reference Range 1M 1M 1M 1M 1M 1M 1M 0.00 0.05 0.00 0.00 0.00 0.00 0.00 00 0.10 0.10 0.08 0.09 0.11 0.09 0.06 00 3.46 3.11 2.94 3.65 2.82 3.13 3.09 00 24.63 10.65 11.14 22.96 7.25 23.68 13.75 00 Page 1 + 418-018:PAGE H-114 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Spec. No. ID B 0037332 11508 B 0037333 11509 B 0037334 11510 B 0037335 11511 B 0037336 11512 B 0037337 11513 B 0037338 11514 Mean S.D. Group 1 Gp Sex Age 1M 1M 1M 1M 1M 1M 1M LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 .004 .014 HDL-DIR MG/GM 0.07 0.06 0.06 0.11 0.07 0.08 .083 .018 CHOLEST MG/GM 3.51 2.85 3.48 3.49 3.73 3.14 3.262 .306 TRIG MG/GM 35.39 11.38 12.80 13.37 24.71 10.42 17.087 8.276 B 0037507 11620 B 0037508 11621 B 0037509 11622 B 0037510 11623 B 0037511 11624 B 0037512 11625 B 0037513 11626 B 0037514 11628 B 0037515 11629 Mean S.D. Group 10 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 10 F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.12 0.09 0.15 0.10 0.11 0.07 0.04 0.09 0.14 .101 .034 3.34 3.57 4.15 3.76 3.41 2.74 2.34 3.50 4.88 3.521 .738 8.79 12.10 13.60 14.72 8.14 12.16 5.38 17.09 20.06 12.449 4.584 Reference Range 0- 0- 00 Page 2 + 0- 0 0- 0 000611 418-018:PAGE H-115 ozmcsC N I INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Spec. No. ID B 0037401 11620 B 0037402 11621 B 0037403 11622 B 0037404 11623 B 0037405 11624 B 0037406 11625 B 0037407 11626 B 0037408 11628 B 0037409 11629 Mean S.D. Group 10 Gp Sex Age 10 M 10 M 10 M 10 M 10 M 10 M 10 M 10 M 10 M LDL-DIR MG/GM 0.00 0.00 0.01 0.01 0.00 0.00 0.04 0.00 0.00 .007 .013 HDL-DIR MG/GM 0.06 0.06 0.10 0.09 0.11 0.09 0.15 0.09 0.13 .098 .029 CHOLEST MG/GM 3.02 3.01 2.79 2.68 3.40 2.93 3.40 2.44 4.20 3.097 .517 TRIG MG/GM 11.72 17.00 8.95 6.73 10.34 14.30 5.22 9.05 14.28 10.843 3.841 B 0037516 11630 B 0037517 11632 B 0037518 11633 B 0037519 11634 B 0037520 11635 B 0037521 11636 B 0037522 11637 B 0037523 11638 B 0037524 11639 Mean S.D. Group 11 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.18 0.12 0.10 0.10 0.10 0.10 0.08 0.09 .109 .031 5.48 4.46 3.54 3.30 3.07 3.21 2.99 3.15 3.65 .875 33.81 16.26 10.71 8.58 8.49 7.34 9.19 9.40 12.973 8.85 Reference Range 00 00 Page 3 + 0 0 00 000612 418-018:PAGE H-116 1 i -Jt JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 Fl LD5 REP2 Species : RAT Accession Sgec. No. B 0037410 11630 B 0037411 11632 B 0037412 11633 B 0037413 11634 B 0037414 11635 B 0037415 11636 B 0037416 11637 B 0037417 11638 B 0037418 11639 Mean S.D. Group 11 Gp Sex Age 11 M 11 M 11 M 11 M 11 M 11 M 11 M 11 M 11 M LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.12 0.12 0.11 0.17 0.12 0.09 0.15 0.17 0.11 .129 .028 CHOLEST MG/GM 3.53 3.23 3.57 2.94 2.87 2.95 3.10 3.46 2.93 3.176 .281 TRIG MG/GM 10.71 16.88 20.07 19.35 5.39 8.15 6.36 8.31 8.36 11.509 5.697 B 0037525 11640 B 0037526 11641 B 0037527 11642 B 0037528 11644 B 0037529 11646 B 0037530 11648 B 0037531 11650 B 0037532 11651 B 0037533 11653 Mean S.D. Group 12 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 12 F 0.00 0.03 0.00 0.00 0.00 0.00 0.00 0.00 .004 .011 0.11 0.19 0.09 0.10 0.13 0.10 0.09 0.09 .113 .034 3.53 3.81 3.12 3.40 3.75 3.24 3.73 3.21 3.474 .271 9.83 14.14 8.43 9.74 14.64 9.54 16.33 6.42 11.134 3.465 Reference Range 0- 0- 0- 00000 Page 4 + 000613 418-018:PAGE H-117 J # V f JL JL V - /K / 200 Girard Street, Suite 200, Gaithersburg, MD 20877 INCORPORATED 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 9 0 5 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 - (215) 443-8710 Date Collected: Date Received: 0 2 / 0 8 / 2 0 0 1 Date Reported: 1 0 / 1 0 / 2 0 0 1 Client No . 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Spec. No. ID B 0037419 11640 B 0037420 11641 B 0037421 11642 B 0037422 11644 B 0037423 11646 B 0037424 11648 B 0037425 11650 B 0037426 11651 B 0037427 11653 Mean S.D. Group 12 Gp Sex Age 12 M 12 M 12 M 12 M 12 M 12 M 12 M 12 M 12 M LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.02 .003 .007 HDL-DIR MG/GM 0.09 0.05 0.12 0.13 0.08 0.12 0.13 0.06 0.08 .096 .03 CHOLEST MG/GM 3.16 2.75 3.23 3.26 2.84 3.37 4.41 3.38 2.87 3.252 .493 TRIG MG/GM 8.54 11.64 6.34 7.98 10.27 9.06 31.92 13.62 6.04 11.712 7.954 B 0037534 11654 B 0037535 11655 B 0037536 11656 Mean S.D. Group 13 13 F 13 F 13 F 0.00 0.00 0.00 0 0 0.11 0.10 0.09 .1 .01 3.30 3.24 4.30 3.613 .595 7.80 11.22 12.20 10.407 2.31 B 0037428 11654 B 0037429 11655 Reference Range 13 M 13 M 0.00 0.02 00 0.11 0.09 00 3.18 3.30 00 7.27 8.74 00 Page 5 + 0C0C14 418-018:PAGE H-l 18 ^ a LmJi A. INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 Fl LD5 REP2 Species : RAT Accession Sgec. No. B 0037430 11656 Mean S.D. Group 13 Gp Sex Age 13 M LDL-DIR MG/GM 0.00 .007 .012 HDL-DIR MG/GM 0.11 .103 .012 CHOLEST MG/GM 3.76 3.413 .306 TRIG MG/GM 5.34 7.117 1.705 B 0037445 11515 B 0037446 11516 B 0037447 11517 B 0037448 11518 B 0037449 11519 B 0037450 11520 B 0037451 11521 B 0037452 11522 B 0037453 11523 B 0037454 11524 B 0037455 11525 B 0037456 11526 B 0037457 11527 B 0037458 11528 Mean S.D. Group 2 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 2F 0.00 0.00 0.00 0.00 0.00 0.02 0.00 0.05 0.00 0.00 0.00 0.00 0.08 .012 .025 0.10 0.13 0.11 0.10 0.10 0.08 0.18 0.14 0.15 0.11 0.08 0.11 0.16 .119 .031 3.80 3.72 3.29 2.98 3.26 2.78 4.52 3.83 4.21 4.06 3.52 3.31 4.96 3.711 .618 17.11 14.02 9.73 7.05 17.42 8.80 26.69 11.21 32.61 16.83 21.48 7.22 29.31 16.883 8.497 Reference Range 0A0 00A Page 6 + 0 0A 00A 000615 418-018:PAGE H-119 4I A I V A ^ INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 9 0 5 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 - (215) 443-8710 Date Collected: Date Received: 0 2 / 0 8 / 2 0 0 1 Date Reported: 1 0 / 1 0 / 2 0 0 1 Client No . 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Sgec. No. B 0037339 11515 B 0037340 11516 B 0037341 11517 B 0037342 11518 B 0037343 11519 B 0037344 11520 B 0037345 11521 B 0037346 11522 B 0037347 11523 B 0037348 11524 B 0037349 11525 B 0037350 11526 B 0037351 11527 B 0037352 11528 Mean S.D. Group 2 Gp Sex Age 2M 2M 2M 2M 2M 2M 2M 2M 2M 2M 2M 2M 2M 2M LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.01 0.00 0.00 0.00 0.04 0.00 .004 .011 HDL-DIR MG/GM 0.12 0.10 0.10 0.10 0.11 0.11 0.07 0.03 0.12 0.09 0.07 0.12 0.10 .095 .026 CHOLEST MG/GM 3.28 2.70 3.08 2.98 3.45 2.79 3.81 4.54 4.07 2.87 2.93 3.16 3.45 3.316 .544 TRIG MG/GM 13.01 5.81 15.74 6.38 18.51 10.59 12.36 7.59 13.11 6.74 9.96 5.25 16.73 10.906 4.419 B 0037459 11529 B 0037460 11530 B 0037461 11534 B 0037462 11536 B 0037463 11537 B 0037464 11538 B 0037465 11543 Mean S.D. Group 3 3F 3F 3F 3F 3F 3F 3F 0.00 0.00 0.02 0.00 0.00 0.00 .003 .008 0.03 0.11 0.08 0.13 0.10 0.08 .088 .034 2.37 2.96 3.06 2.88 2.80 2.64 2.785 .248 10.24 13.20 8.90 9.99 16.11 8.74 11.197 2.893 Reference Range 00 00 Page 7 + 0 0 00 OCOCiS 418-018:PAGE H-120 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Spec. No. ID B 0037353 11529 B 0037354 11530 B 0037355 11534 B 0037356 11536 B 0037357 11537 B 0037358 11538 B 0037359 11543 Mean S.D. Group 3 Gp Sex Age 3M 3M 3M 3M 3M 3M 3M LDL-DIR MG/GM 0.00 0.00 0.00 0.08 0.00 0.00 0.00 .011 .03 HDL-DIR MG/GM 0.05 0.12 0.10 0.12 0.14 0.10 0.11 .106 .028 CHOLEST MG/GM 3.16 2.83 3.09 2.68 3.14 2.94 3.12 2.994 .184 TRIG MG/GM 13.12 11.65 9.38 7.54 15.19 9.76 15.43 11.724 3.015 B 0037466 11558 B 0037467 11559 B 0037468 11561 B 0037469 11563 B 0037470 11564 B 0037471 11565 B 0037472 11567 B 0037473 11568 B 0037474 11569 B 0037475 11570 B 0037476 11571 Mean S.D. Group 5 5F 5F 5F 5F 5F 5F 5F 5F 5F 5F 5F 0.00 0.00 0.07 0.00 0.00 0.00 0.00 0.03 0.00 0.09 0.00 .017 .033 0.14 0.09 0.12 0.16 0.06 0.10 0.16 0.10 0.05 0.12 0.13 .112 .036 4.02 3.22 3.71 4.56 2.36 3.55 4.31 3.06 3.17 3.56 4.43 3.632 .667 21.52 11.76 11.35 25.84 9.81 10.87 22.76 7.64 27.62 9.52 29.93 17.147 8.389 Reference Range 00 00 Page 8 + 0 0 00 000617 418-018:PAGE H-121 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 FI LD5 REP2 Species: RAT Accession Sgec. No. B 0037360 11558 B 0037361 11559 B 0037362 11561 B 0037363 11563 B 0037364 11564 B 0037365 11565 B 0037366 11567 B 0037367 11568 B 0037368 11569 B 0037369 11570 B 0037370 11571 Mean S.D. Group 5 Gp Sex Age 5M 5M 5M 5M 5M 5M 5M 5M 5M 5M 5M LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.03 0.00 0.03 0.00 .005 .012 HDL-DIR MG/GM 0.11 0.11 0.09 0.06 0.08 0.07 0.13 0.09 0.05 0.11 0.11 .092 .025 CHOLEST MG/GM 4.31 2.93 3.38 3.55 3.57 3.58 3.63 3.45 3.05 3.43 4.34 3.565 .435 TRIG MG/GM 23.45 19.3 5 19.76 18.74 9.09 11.62 12.33 8.43 30.72 7.14 22.41 16.64 7.469 B 0037477 11574 B 0037478 11575 B 0037479 11576 B 0037480 11578 B 0037481 11580 B 0037482 11582 B 0037483 11583 B 0037484 11584 Mean S.D. Group 6 6F 6F 6F 6F 6F 6F 6F 6F 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.10 0.12 0.05 0.09 0.10 0.10 0.09 0.11 .095 .021 2.98 3.34 3.42 2.72 2.73 3.14 3.26 3.82 3.176 .369 7.73 20.40 9.36 9.82 6.93 10.80 14.62 15.38 11.88 4.564 Reference Range 0- 0- 0- 00000 Page 9 + 000618 418-018:PAGE H-122 1 JL J L K * / K-y 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NC OR P O R A T E D 301-921-0168 Fax:301-977-0433 client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported:. 10/10/2001 Client No . 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Spec. No. ID B 0037371 11574 B 0037372 11575 B 0037373 11576 B 0037374 11578 B 0037375 11580 B 0037376 11582 B 0037377 11583 B 0037378 11584 Mean S.D. Group 6 Gp Sex Age 6M 6M 6M 6M 6M 6M 6M 6M LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.12 0.11 0.14 0.12 0.08 .114 .022 CHOLEST MG/GM 3.78 3.07 3.01 2.74 3.17 3.154 .384 TRIG MG/GM 14.76 6.69 6.64 7.02 9.75 8.972 3.484 B 0037485 11592 B 0037486 11593 B 0037487 11597 Mean S.D. Group 7 7F 7F 7F 0.00 0.00 0 0 0.11 0.10 .105 .007 4.12 3.88 4 .17 19.32 15.71 17.515 2.553 B 0037379 11592 B 0037380 11593 B 0037381 11597 Mean S.D. Group 7 7M 7M 7M 0.00 0 0 0.11 .11 0 3.25 3.25 0 8.48 8.48 0 Reference Range 0- 000 Page 10 + 00 00 0C013 418-018:PAGE H-123 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No . 1028 Study: 418018 Fl LD5 REP2 Species : RAT Accession Spec. No. ID B 0037488 11600 B 0037489 11601 B 0037490 11602 B 0037491 11603 B 0037492 11604 B 0037493 11605 B 0037494 11606 B 0037495 11607 B 0037496 11608 B 0037497 11609 Mean S.D. Group 8 B 0037382 11600 B 0037383 11601 B 0037384 11602 B 0037385 11603 B 0037386 11604 B 0037387 11605 B 0037388 11606 B 0037389 11607 B. 0037390 11608 B 0037391 11609 Mean S.D. Group 8 Gp Sex Age 8F 8F 8F 8F 8F 8F 8F 8F 8F 8F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.01 0.00 0.00 0.00 0.00 .001 .003 HDL-DIR MG/GM 0.08 0.13 0.13 0.14 0.12 0.12 0.09 0.11 0.06 0.11 .109 .025 CHOLEST MG/GM 3.64 3.91 3.84 3.22 4.04 3.29 4.49 4.80 2.82 3.67 3.772 .591 TRIG MG/GM 11.24 21.85 17.00 8.90 24.28 4.49 23.10 19.22 7.97 12.59 15.064 6.972 8M 8M 8M 8M 8M 8M 8M 8M 8M 8M 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.11 0.09 0.10 0.08 0.11 0.13 0.05 0.06 0.09 0.09 .091 .024 3.85 3.65 3.36 3.30 3.00 3.31 3.73 3.38 2.75 3.12 3.345 .337 15.00 21.52 9.48 18.07 8.62 9.32 20.78 13.09 4.53 15.49 13.59 5.602 Reference Range 0- 0- 00 Page 11 + 0- 0 0- 0 060620 418-018:PAGE H-124 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044- (215) 443-8710 Date Collected: Date Received: 02/08/2001 Date Reported: 10/10/2001 Client No.. 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Sgec. No. B 0037498 11611 B 0037499 11612 B 0037500 11613 B 0037501 11614 B 0037502 11615 B 0037503 11616 B 0037504 11617 B 0037505 11618 B 0037506 11619 Mean S.D. Group 9 Gp Sex Age 9F 9F 9F 9F 9F 9F 9F 9F 9F LDL-DIR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 HDL-DIR MG/GM 0.16 0.15 0.15 0.12 0.10 0.10 0.10 0.10 0.08 .118 .029 CHOLEST MG/GM 3.90 3.51 4.04 3.29 3.93 3.67 3.60 3.83 3.72 3.721 .234 TRIG MG/GM 15.82 13.82 15.87 7.98 10.96 20.01 11.90 21.01 23.13 15.611 5.025 B 0037392 11611 B 0037393 11612 B 0037394 11613 B 0037395 11614 B 0037396 11615 B 0037397 11616 B 0037398 11617 B 0037399 11618 B 0037400 11619 Mean S.D. Group 9 9M 9M 9M 9M 9M 9M 9M 9M 9M 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 0.10 0.10 0.12 0.09 0.03 0.13 0.08 0.15 0.14 .104 .036 3.17 2.86 3.60 3.31 3.18 3.61 3.09 3.28 2.77 3.208 .287 16.14 7.61 23.50 16.50 15.77 12.62 10.08 29.27 7.90 15.488 7.183 Reference Range 0- 0- 0 0 o y0 Approved -- -- '>CL__ Page 12 of 12 00 Date 000621 418-018.`PAGE H-125 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 800-237-2815 QA AUDIT REPORT The report listed below was reviewed for compliance with the FDA Good Laboratory Practices and with EPA Good Laboratory Practices. The final report and all associated raw data were reviewed for accuracy and consistency and the findings were reported to management. The methods used were the methods described and the report accurately reflects the data. Therefore, these studies were done in compliance with the FDA and EPA Good Laboratory Practices. Franklin B. Newman QA Auditor SPONSOR: REPORT TYPE cM.LQK / A & 5 ______ STDDY: H H Q i t _____________ AUDIT DATE REPORT TO MANAGEMENT -JIL__ AUDIT # 91lift 's,/o t TO b / '/(trie. DATE SUBMITTED TO STUDY DIRECTOR r ^//MANAGEMENT 0/ 000622 APPENDIX I BIOANALYTICAL REPORTS (PRIMEDICA WORCESTER, SPONSOR AND SRI) 000623 418-018:PAGE 1-1 FINAL REPORT VALIDATION OF A GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC METHOD FOR THE ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR PRIMEDICA ARGUS STUDY 418-018 Primedica-Worcester Project Number: PABX-BVA PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 000624 418-018-.PAGE 1-2 O P rimedica FINAL REPORT VALIDATION OF A GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC METHOD FOR THE ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR PRIMEDICA ARGUS STUDY 418-018 Primedica-Worcester Project Number: PABX-BVA Submitted to: PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 Submitted by: Primedica-Worcester 57 Union Street Worcester, MA 01608 Primedica-Worcester Report # PABX-BVA-01 -62 Page J. of 184 Issue Date: November 7, 2001 M * Mark A. Netsch / Date Project Scientist Department of Analytical Chemistry Primedica-Worcester 00062^ 418-018:PAGE 1-3 OPrimedica Primedica-Worcester Project number: PABX-BVA Page 2 Final Report_______________________________ For Primedica Argus Study Number: 418-018 COMPLIANCE STATEMENT This project was conducted in compliance with the Good Laboratory Practice Regulations, 21CFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision/ recommendation on compliance with principles of good laboratory practice. Official Journal of European Communities: Legislation 32 (No. L 315; 28 October): 1-17. There were no deviations from the aforementioned standards, which affected the quality or integrity of the project or the interpretation of the results in this report. Mark A. Netsch / Date Project Scientist 0G0626 418-018-.PAGE 1-4 S P rimedica Primedica-Worcester Project number: PABX-BVA Page 3 Final Report_______________________________ For Primedica Argus Study Number: 418-018 QUALITY ASSURANCE STATEMENT The following are the inspection dates and report dates of QAU audit/inspections for Validation Of A Gas Chromatographic-Mass Spectrometric Method For The Analysis Of Mevalonic Acid In EDTA Rat Plasma For Primedica Argus Study 418-018, Project Number PABX-BVA. Critical Phases 1. Laboratory Procedures 1. Raw Data 2. Draft Final Report Date Inspected 10/19/2000 11/07/2000 03/26/2001 Date Report Submitted to Project Scientist Management 10/20/2000 10/23/2000 04/06/2001 04/06/2001 03/26/2001 04/04/2001 The Final Report for Validation of A Gas Chromatographic-Mass Spectrometric Method For The Analysis Of Mevalonic Acid In EDTA Rat Plasma For Primedica Argus Study 418-018, report number PABX-BVA-01-62, was reviewed for compliance with the Good Laboratory Practice Regulations, 21CFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision/ recommendation on compliance with principles of good laboratory practice. Official Journal of European Communities: Legislation 32 (No. L 315; 28 October): 1-17, on 11/7/01. The results as presented accurately reflect the raw data. Quality Assurance uditor Date 000627 418-018:PAGE 1-5 O Primedica Primedica-Worcester Project number: PABX-BVA Page 4 Final Report_______________________________ For Primedica Argus Study Number: 418-018 TABLE OF CONTENTS Page No. COMPLIANCE STATEMENT................................................................................. 2 QUALITY ASSURANCE STATEM ENT.......................................... 3 LIST OF TABLES AND APPENDICES....................................................................5 CONTRIBUTING PERSONNEL............................................................................. 7 ANALYTICAL REFERENCE STANDARD CHARACTERIZATION/STABILITY....8 ARCHIVAL STO R AGE.......................................................................................... 9 A B S T R A C T ........................................................................................................... 10 INTRODUCTION............................................................................................................... 11 EXPERIMENTAL DESIGN.............................................................................................. 11 Overview....................................... ................................................................................. 11 Materials and Methods.....................................................................................................11 General Comments.........................................................................................................11 Validation Design.............................................................................................................11 Validation Description.................................................................................... 11 Parameters Evaluated....................................................................................................12 RESULTS AND DISCUSSION......................................................................................... 14 Validation Results.............................................................................................................14 Water/Plasma Extraction Comparison......................................................................... 14 Accuracy........................................................................................................................ 15 Precision........................................................................................................................ 15 Dilution QC................................................................................................................... 16 Recovery........................................................................................................................ 16 Linearity........................................................................................................................ 16 Specificity...................................................................................................................... 16 Stability......................................................................................................................... 17 CONCLUSIONS.................................................................................................................. 18 000628 G P rimedica 418-018:PAGE 1-6 LIST OF TABLES, FIGURES AND APPENDICES Page No. TEXT TABLE 1 Validation Course.......................................................................... ........................ 12 FIGURE 1 Day 1 Validation Calibration Curve...................................................................... 20 TABLE 1 Summary of Interpolated Mevalonic Acid Lactone QC Standard Concentrations with Inter and Intra-Day Summary Statistics........................................................ 22 TABLE 2 Summary of Back-Calculated Mevalonic Acid Lactone Calibration Standard Concentrations........................................................................................................ 24 TABLE 3 Mevalonic Acid Lactone Extraction Data & Recovery Calculations....................25 TABLE 4 Internal Standard Extraction Data & Recovery Calculations...............................26 TABLE 5 Summary of Least-Squares Linear Regression Constants and Analysis Dates.... 27 TABLE 6 Summary of QC Standard 72-Hour Room Temperature Stability in Matrix........ 28 TABLE 7 Summary of QC Standard Freeze/Thaw (3 Cycles) Stability in Matrix............... 29 TABLE 8 Summary of Interpolated Mevalonic Acid Lactone Dilution QC Concentrations for Dilution Validation............................................................................................ 30 000629 G P rimedica 418-018:PAGE 1-7 LIST OF TABLES, FIGURES AND APPENDICES - CONCLUDED Page No. TABLE 9 6-Day Room Temperature Final Extract Stability Back-Calculated Calibration Standard Concentrations........ ................................................................................ 31 TABLE 10 6-Day Room Temperature Final Extract Stability Quality Control Standard Concentrations........................................................................................................ 32 TABLE 11 Summary of Control Blank EDTA Rat Plasma Concentrations............................33 TABLE 12 Summary of Unique Lots of EDTA Plasma Concentrations.................................34 TABLE 13 Summary of Internal Standard Peak Areas............................................................ 35 TABLE 14 Summary of Plasma QC Standard Concentrations with Inter- and Intra-Day Summary Statistics................................................................................................. 37 APPENDIX A Laboratory Method................................................................................................ 39 APPENDIX B Representative Chromatograms from Validation Day 1 ...................................... 52 0C0C30 418-018:PAGE 1-8 O Primedica Primedica-Worcester Project number: PABX-BVA Page 7 Final Report_______________________________ For Primedica Argus Study Number: 418-018 CONTRIBUTING PERSONNEL Project Scientist...................... Research Associate................. Director, Analytical Chemistry Report Coordinator................. . Mark A. Netsch, B.A. .... Heidi Gagnon, B.S. James A. Jersey, Ph.D. ....... Brenda L. Brooks 000631 418-018:PAGE 1-9 O P rimedica Primedica-Worcester Project number: PABX-BVA Page s Final Report_______________________________ For Primedica Argus Study Number: 418-018 ANALYTICAL REFERENCE STANDARD CHARACTERIZATION/STABILITY Analytical Reference Standard: Common Name: Physical Description: Lot Number: Storage Conditions: Expiration Date: Date Received: Amount Received: Supplier: Internal Standard: Common Name: Physical Description: Lot Number: Storage Conditions: Expiration Date: Date Received: Amount Received: Supplier: DL-Mevalonic Acid Lactone Mevalonic Acid Lactone or Mevalonolacetone or MVL White crystals 47300/1 50498 53 C, desiccate, store under nitrogen September 13, 2002 (Assigned by Primedica) September 13, 2000 5 grams Aldrich Chemical Co. DL-Mevalonolacetone-4,4,5,5-d4 MVL-d Clear liquid U-295 225C September 11, 2002 (Assigned by Primedica) September 11, 2000 0.1 gram Cambridge Isotope Laboratories Characterization and Stability: The characterization of the analytical standard and internal standard is the responsibility of the Supplier, as is the method of synthesis, fabrication or derivation and stability determination. 000632 418-018.-PAGE I - 10 O P rimedica Primedica-Worcester Project number: PABX-BVA Page 9 Final Report _____________ __________ For Primedica Argus Study Number: 418-018 ARCHIVAL STORAGE Archival Storage: The original Final Report and raw data will be maintained for a minimum period of five years following submission of the final report in the Primedica-Worcester Archives in Worcester, MA. After five years the Sponsor will be contacted for disposition instructions. Archival material will be indexed by Primedica-Worcester Report #PABX-BVA-01-62. 000633 418-018:PAGE 1-11 G Primedica Primedica-Worcester Project number: PABX-BVA Page 10 Final Report______________________ For Primedica Argus Study Number: 418-018 ABSTRACT: A procedure has been developed and validated for the determination of mevalonic acid in EDTA rat plasma. The procedure involves the conversion of mevalonic acid (MVA) to mevalonic acid lactone (MVL) and the analysis of a liquid/liquid extract by gas chromatography coupled with mass spectrometry (GC/MS) using positive chemical ionization (PCI). A mathematical factor to convert sample concentrations of MVL to MVA is included in the Laboratory Method in Appendix A. Calibration and quality control standards are prepared in water since mevalonic acid is a naturally occurring compound in plasma and concentrations vary with diet. The internal standard, MVL-cL, was used for quantification. The results for the validation indicate that the method is sufficiently linear, specific, reproducible and accurate to support the analysis of mevalonic acid in EDTA rat plasma samples. 000634 418-018:PAGE 1-12 O Primedica Primedica-Worcester Project number: PABX-BVA Page 11 Final Report_______________________________ For Primedica Argus Study Number: 418-018 INTRODUCTION: The objective of this study was to develop and validate an analytical method for determining levels of mevalonic acid (MVA) in EDTA rat plasma. EXPERIMENTAL DESIGN: Overview: The assay described in this report employed the conversion of mevalonic acid (MVA) to mevalonic acid lactone (MVL) and the analysis of liquid/liquid EDTA rat plasma samples by GC/MS using positive chemical ionization. The internal standard, MVL-tL*, was used for quantitation. The validated assay covered the concentration range of 10.0 ng/mL to 250 ng/mL of MVL in EDTA rat plasma using 100 pL sample volumes. Materials and Methods: Refer to Appendix A for a detailed description of materials and methods employed during the course of this validation study. In addition, quality control (QC) standards were prepared in water at the lower limit of quantiation (LLOQ QC, 10.0 mg/mL) using the same stock solutions used for the water QC standards. A set of QC standards (Low, Mid and High) was prepared in rat plasma using the same stock standards and dilution procedure used for the water QC standards. One set of solution calibration standards was prepared to mimic final extract concentration assuming 100% recovery using the same stock standards used for the calibration standards. General Comments: Representative raw chromatographic data from validation day 1 are provided in Appendix B. Validation Design: Validation Description: The assay was comprised of five analytical runs. Three of the five analytical runs were comprised of bracketing seven-point water matrix calibration curves with six replicates of water matrix quality control (QC) samples at four concentrations, six replicates of EDTA rat plasma matrix QC samples at three concentrations and six replicates of rat matrix control samples (EDTA rat plasma with internal standard). Multiple water matrix blanks and water matrix control samples (matrix with internal standard) were analyzed in each 0G0635 418-018:PAGE 1-13 S P rimedica Primedica-Worcester Project number: PABX-BVA Page 12 Final Report__________________ For Primedica Argus Study Number: 418-018 run. An analysis of the method recovery was included in the third analytical run, which consisted of an extracted calibration curve and a solvent calibration curve. The analysis of ffeeze/thaw (F/T) stability and room temperature (RT) stability, both in water and matrix, was included in the fourth analytical run. The fifth analytical run was a re-injection of the extracts from the first run to evaluate the final extract stability when stored at room temperature. Text Table 1 lists each validation analytical run and the assay performance parameters that were evaluated for the study. TEXT TABLE 1 Validation Course Validation Run Identification Parameters Evaluated Day 1 Precision, accuracy, linearity, and water/plasma equivalency Day 2 Precision, accuracy, linearity, specificity, and water/plasma equivalency Day 3 Precision, accuracy, linearity, recovery, dilution accuracy, precision, and water/plasma equivalency Day 4 Freeze/thaw and room temperature stability Day 5 6-day room temperature final extract stability Parameters Evaluated: Mevalonic acid is a naturally occurring compound in plasma. The concentration levels found in blank plasma were anticipated to be above the validated lower limit of quantitation (LLOQ) of 10 ng/mL. This level of background concentration would significantly skew the results of plasma QC's at the lower concentration levels. Therefore, calibration and QC standards were prepared in water matrix to evaluate precision, accuracy and linearity of the method. The second set of QC's was prepared at three concentration levels in EDTA rat plasma to evaluate the ability of the method to extract and quanitate MVL equally from water or plasma matrices. The plasma QC's were also used to evaluate matrix stability. 000638 418-018:PAGE 1-14 O P rimedica Primedica-Worcester Project number: PABX-BVA Page 13 Final Report_______________________________ For Primedica Argus Study Number: 418-018 Water/plasma extraction equivalency was evaluated by the analysis of six replicates of each QC standard concentration levels in both water and plasma. Mean peak area counts of the isotopically labeled mevalonic acid lactone internal standard were calculated. The difference between the water and plasma mean peak areas were expressed as percent difference. The acceptable limits were 10% difference. The isotopically labeled internal standard was used to evaluate extraction equivalency because it is chemically identical to mevalonic acid lactone and is not present in blank plasma. Therefore, the internal standard mimics the extraction behavior of mevalonic acid lactone but has no naturally occurring background concentration to interfere with the results. Water/plasma extraction equivalency was also evaluated by the six replicates of High QC standards in both water and plasma. The endogenous concentration of mevalonic acid was low compared to the high QC concentration and did not make a significant contribution to the concentration results. Mean concentrations of mevalonic acid lactone were calculated and expressed as percent bias compared to the nominal concentration. The present bias values from the high QC in plasma were compared with the water QC and expressed as percent difference. The acceptable limits were 10% difference. Inter- and intra-day accuracy and precision of the method were evaluated at four concentrations by analyzing six replicates of the QC standards prepared in water over the course of three validation batches. Accuracy results are reported as % bias. Method precision for QC standards is expressed as % variance. Inter-day method precision and accuracy were also evaluated for calibration standards. Dilution accuracy and precision was measured by analyzing six replicate dilutions of a QC standard diluted 1:5 water. Intra-day precision and accuracy was evaluated and reported as % RSD and % bias, respectively. Recovery of the assay was evaluated by comparing the absolute peak areas in extracted standards to those obtained from solvent standards prepared to mimic final extract concentrations assuming 100% recovery. Linearity of the method was evaluated by visual inspection of the calibration curve and examination of the correlation coefficient (r) for each standard curve. Freeze-Thaw (F/T) stability in both water and plasma matrices were evaluated by analyzing six replicates of quality control samples prepared at two levels, which were subjected to three F/T cycles, prior to their extraction and analysis. 000637 418-018:PAGE 1-15 G Primedica Primedica-Worcester Project number: PABX-BVA Page 14 Final Report ____________________________ For Primedica Argus Study Number: 418-018 Room temperature stability in both water and plasma matrices were demonstrated by analyzing six replicates of QC standards prepared at two levels which were left standing at room temperature for seventy-two hours prior to their extraction and analysis. Six-day room temperature stability of final extracts was evaluated by re-analyzing the day-one validation extracts consisting of duplicate calibration curves, six replicates of QC standards in both water and plasma matrices. Assay specificity with respect to endogenous compounds was evaluated by assaying six unique plasma lots from untreated rats. RESULTS AND DISCUSSION: Validation Results: Water/PIasma Extraction Equivalency: The ability of the method to extract and quantitate mevalonic acid equivalently from water or plasma matrices was evaluated. The equivalence was evaluated by the analysis of six replicates of each of three QC standard levels in water and plasma during the first three validation runs. Mean peak area cotints of the internal standard (isotopically labeled mevalonic acid lactone) found for the plasma matrix QC's during each run were compared to the results for water matrix QC's. The difference was expressed as % difference. Percent difference values for the first three validation runs were 0.5%, 0.5% and 4.4%, respectively. Refer to Table 13 for a complete summary of results. The comparison was also evaluated by the analysis of six replicates of high-QC standards in water and plasma during the first three validation runs. At this concentration level (200 ng/mL) the endogenous level of mevalonic acid was not a significant interference. Mean concentrations of found for the plasma matrix High-QC's during each run was compared to the results for water matrix High-QC's. The results were expressed as % difference. Percent difference values for the first three validation runs were -2.0%, 0.5% and 0.5%, respectively. Refer to Table 14 for a complete summary of results. The percent difference results were all within the acceptable limits of 10%. The extraction of mevalonic acid lactone from water and plasma are equivalent. Therefore, GG063& 418-018:PAGE 1-16 G P rim edica Primedica-Worcester Project number: PABX-BVA Page 15 Final Report_______________________________ For Primedica Argus Study Number: 418-018 the calibration curve and quality control sample prepared in water can be used to quantitate mevalonic acid lactone concentration levels in plasma. Accuracy: Inter-day and intra-day accuracy were acceptable for the assay of Mevalonic acid lactone in water over the concentration range of 10.0 ng/mL to 250 ng/mL. Intra-day accuracy was evaluated by the analysis of six replicates of each of four QC standard levels in water during the first three validation runs. Mean concentrations found during each run were compared to theoretical nominal concentrations and the difference relative to theoretical nominal concentrations was expressed as % bias. Percent bias values for the lower limit of quantitation (LLOQ-QC, 10 ng/mL), low-QC (25 ng/mL), middle-QC (70 ng/mL), and high-QC (200 ng/mL) standards ranged from --4.8% to 0.1% over the three analytical runs. Refer to Table 1 for a complete summary of results. Inter-day accuracy was evaluated by the analysis of six replicates of each of four QC standard levels in water during the first three validation runs. Mean concentrations across these runs were compared to theoretical nominal concentrations and the difference was expressed as % bias. Percent bias values ranged from -3.6% to -1.4% across concentrations. Refer to Table 1 for a complete summary of results. The inter-day accuracy statistics for the back-calculated calibration standard concentrations are also represented by % bias. The % bias values for the calibration standards ranged from -1.6% to 1.7%. See Table 2 for a complete summary of results. Precision: Inter-day and intra-day precision was acceptable for the assay of mevalonic acid lactone in water over the concentration range of 10.0 ng/mL to 250 ng/mL. Intra-day precision was evaluated by the analysis of six replicates of each of four QC standard levels in water during the first three validation runs. The intra-assay variance estimate of the interpolated concentrations for the replicate QC standards ranged from 1.3% to 4.5% across concentrations. Refer to Table 1 for a complete summary of results. Inter-day precision was evaluated by the analysis of six replicates of each of four QC standard levels in water during the first three validation runs. The inter-assay variance estimate of the interpolated concentrations for the replicate QC standards ranged from 0.4% to 1.2% across concentrations. Refer to Table 1 for a complete summary of results. fci;c&39 418-018:PAGE 1-17 G P rimedica Primedica-Worcester Project number: PABX-BVA Page 16 Final Report_______________________________ For Primedica Argus Study Number: 418-018 The inter-day precision statistics for the back-calculated calibration standards ranged from 1.6% to 3.0% relative standard deviation (RSD) across concentrations and analytical runs. See Table 2 for a complete summary of results. Dilution QC: The dilution QC results were acceptable. Six replicates of the dilution QC with a pre dilution concentration of 200 ng/mL, were diluted with water by a factor of 5 prior to extraction and analysis. The mean concentration found was compared to the theoretical nominal concentration (40 ng/mL) and the difference relative to the theoretical nominal concentration was expressed as % bias. The result was -1.8% bias with an RSD of 3.0%. See Table 8 for a complete summary of results. Recovery: Recovery values were calculated by comparison of mean duplicate spiked and extracted calibration standard peak areas in water to the peak areas obtained for solution standards prepared to mimic final extract concentrations assuming 100% recovery. Recovery ranged from 32.9% to 39.6% for mevalonic acid lactone and from 33.6% to 42.0% for the internal standard across concentrations. Refer to Tables 3 and 4 for a complete summary of results. Linearity: The assay was linear for mevalonic acid lactone within the calibration range of 10.0 ng/mL to 250 ng/mL. y Figure 1 shows the 1/X weighted least-squares linear regression plot with the actual spiked calibration standard data points for one of the validation runs. Linearity was also indicated by the correlation coefficients (r) obtained, which were greater than or equal to 0.998 for each of the first three validation runs. Refer to Table 5 for a summary of regression constants. Specificity: Mevalonic acid is a naturally occurring compound in plasma. The background concentration level in the plasma QC's was evaluated by the analysis of six replicates of control blank EDTA rat plasma matrix during the first three validation runs. The inter-assay mean concentration found was 16.6 ng/mL. Refer to Table 11 for complete results for the control blanks of plasma. 000640 418-018:PAGE 1-18 G P rimedica Primedica-Worcester Project number: PABX-BVA Page 17 Final Report_______________________________ For Primedica Argus Study Number: 418-018 The variation of background concentration levels in EDTA rat plasma was evaluated by the analysis of six unique lots of plasma during validation Day 2. The result was a mean concentration of 15.8 ng/mL. See Tables 12 for complete results of the six unique lots of plasma. Stability: The acceptable criteria for stability evaluations were a mean bias of <15% from the theoretical values except for the lowest calibration, Low-QC and LLOQ QC standards, where the criteria were <20%. Due to the presence of background concentrating of mevalonic acid lactone the theoretical values used for the Plasma QC standards were the mean concentrations obtain during the same analytical run from the analysis of six control Plasma QC standards. Room temperature matrix stability was evaluated by analyzing six replicates of quality control samples prepared at two levels, in both water and EDTA rat plasma matrices, which were exposed to room temperature for 72-hours prior to their extraction and analysis. The values for % bias were within the acceptance criteria for both concentrations for the 72-hour room temperature stability assessment. Refer to Table 6 for a complete summary of the results. Freeze/thaw matrix stability was evaluated by analyzing six replicates of QC standards prepared at two levels, in both water and EDTA rat plasma matrices that were subjected to four freeze/thaw cycles. The values for % bias were within the acceptance criteria. Refer to Table 7 for a complete summary of the results. Six-day room temperature stability of final extracts was evaluated by re-analyzing the duplicate calibration curves and the six replicates of QC standards, in both water and EDTA rat plasma matrices, from the first day of the validation. The solvent in the extracts evaporated from the vials during the storage period. More solvent was added to each vial to reconstitute the extracts. Results from the re-analyses demonstrated that the extracts were stable for a minimum period of six days when stored at room temperature. See Tables 9 and 10 for the final extract stability results. Studies of long-term storage stability of mevalonic acid in matrix at -20C are ongoing. 000541 418-018:P A G E 1-19 O P r im e d ic a Primedica-Worcester Project number: PABX-BVA Page 18 Final Report____________________________________ For Primedica Argus Study Number: 418-018 CONCLUSIONS: Overall, the results for the validation indicated that the method was sufficiently linear, specific, reproducible and accurate to support EDTA rat plasma sample analyses for mevalonic acid content. 000642 Primedica-Worcester Project number: PA B X -B V A 418-018:P A G E 1-20 Page 19 FIGURE 000643 Primedica-Worcester Project number: PA BX -BV A FIGURE 1 Day I Validation Calibration Curve 418 -0 1 8 :P A G E 1-21 Page 20 1.8 1.6 1.4 1.2 - r 1 0.8 - r 0.6 0.4 0.2 0 Mevalonic Acid in Rat Plasma Validation Day 1 50 100 150 200 250 300 Concentrationinng/mL Rat Plasma PeakArea Ratio OG0644 Primedica-Worcester Project number: PA BX -BV A 418-018:P A G E 1-22 P a g e 21 TABLES 00064 Primedica-Worcester Project number: PABX-BVA 418-018:P A G E 1-23 Page 22 TABLE 1 SUMMARY OF INTERPOLATED MEVALONIC ACID LACTONE QC STANDARD CONCENTRATIONS WITH INTER- AND INTRA-DAY SUMMARY STATISTICS Day 1 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean n % Bias Theoretical Nominal Concentrations (ng/mL) 10.0 25.0 70,0 200 9.63 8.91 9.51 9.38 9.75 9.92 23.6 25.0 25.0 23.9 24.0 23.9 70.1 68.7 67.8 68.3 67.4 69.8 202 200 196 196 195 199 9.52 6 -4.8% 24.2 6 -3.2% 68.7 6 -1.9% 198 6 -1.0% Day 2 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean n % Bias Theoretical Nominal Concentrations (ng/mL) 10.0 25.0 70.0 200 9.87 9.29 10.3 9.07 9.10 10.2 24.4 24.8 24.1 23.8 24.8 24.5 69.9 68.1 68.4 68.2 68.0 67.3 202 195 195 193 194 195 9.64 6 -3.6% 24.4 6 -2.4% 68.3 6 -2.4% 196 6 -2.0% 00064 Primedica-Worcester Project number: P A B X -B V A 418-018:PA G E 1-24 Page 23 TABLE 1 (Concluded) SUMMARY OF INTERPOLATED MEVALONIC LACTONE ACID QC STANDARD CONCENTRATIONS WITH INTER- AND INTRA-DAY SUMMARY STATISTICS Day 3 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean n % Bias Theoretical Nominal Concentrations (ng/mL) 10.0 25.0 70.0 200 9.49 10.1 9.49 9.96 9.84 NA 23.9 70.6 25.2 70.1 25.4 70.6 24.6 70.0 24.8 70.0 23.5 69.0 198 197 200 198 199 188 9.78 5 -2.2% 24.6 6 -1.6% 70.1 6 0.1% 197 6 -1.5% Intra-assay variance estimate Inter-assay variance estimate 4.5% 1.2% 2.5% 0.7% 1.3% 1.2% 1.8% 0.4% Inter-day Mean n % Bias 9.64 17 -3.6% 24.4 18 -2.4% 69.0 18 -1.4% 197 18 -1.5% N A = Not Available. This sample was accidentally spilled during preparation G00S47 Primedica-Worcester Project number: PABX-BVA 418-018:P A G E 1-25 Page 24 TABLE 2 SUMMARY OF BACK-CALCULATED MEVALONIC ACID LACTONE CALIBRATION STANDARD CONCENTRATIONS Day 1 Day 2 Day 3 10.0 9.90 10.1 9.84 10.3 10.3 9.54 Theoretical Nominal Concentrations (ng/mL) 20.0 30.0 50.0 90.0 150 20.0 20.0 30.7 30.0 50.6 49.7 89.5 88.1 151 144 19.5 29.6 50.4 90.8 19.3 31.6 49.2 91.7 152 146 19.7 20.9 30.5 30.6 51.7 47.8 91.7 84.8 147 146 250 254 254 250 250 261 251 Mean Std. Dev. n %RSD % Bias 10.0 0.296 6 3.0% 0.0% 19.9 0.562 6 2.8% -0.5% 30.5 0.681 6 2.2% 1.7% 49.9 1.34 6 2.7% -0.2% 89.4 2.66 6 3.0% -0.6% 148 3.14 6 2.1% -1.6% 253 4.18 6 1.6% 1.3% GG0648 Primedica-Worcester Project number: PA B X -B V A 418-018:P A G E 1-26 Page 25 TABLE 3 MEVALONIC ACID LACTONE EXTRACTION DATA & RECOVERY CALCULATIONS Std 1 Std 2 Std 3 Std 4 Std 5 Std 6 Std 7 Extracted Standards Solution Standards Mean Resoonse Areas Response Areas 417.5 785 1057.5 1896 3144 4693.5 8433 1066 1981 2963 4975 8678 14279 25380 Mean Std. D ev. % RSD % Recover/ 39.2% 39.6% 35.7% 38.1% 36.2% 32.9% 33.2% 36.4% 0.0327 7.4% 000649 Primedica-Worcester Project number: PA B X -B V A 418-018:P A G E 1-27 Page 26 TABLE 4 INTERNAL STANDARD EXTRACTION DATA & RECOVERY CALCULATIONS Extracted Standards Solution Standards Mean Resoonse Area Resoonse Area Std 1 Std 2 Std 3 Std 4 Std 5 Std 6 Std 7 6142 5876 5365 6004 5657 5117.5 5274.5 15019 14959 15158 14283 14739 15226 15286 Mean Std. Dev. % RSD % Recovery 40.9% 39.3% 35.4% ' 42.0% 38.4% 33.6% 34.5% 37.7% 0.0327 8.7% CCO60O Primedica-Worcester Project number: PA B X -B V A 418-018:P A G E 1-28 Page 27 TABLE 5 SUMMARY OF LEAST-SQUARES LINEAR REGRESSION CONSTANTS AND ANALYSIS DATES Day 1 Day 2 Day 3 Day 4 Day 5 SloDe 0.0065514 0.0063508 0.0062253 0.0062800 0.0065156 Intercent 0.0113750 0.0082756 0.0067910 0.0060925 0.0112850 Correlation C oefficient fr) 0.99975 0.99939 0.99884 0.99891 0.99981 Analysis Date 10/17/00 10/18/00 10/19/00 10/20/00 10/23/00 000651 Primedica-Worcester Project number: PABX-BVA 418-018:P A G E 1-29 Page 28 TABLE SUMMARY OF QC STANDARD 72-HOUR ROOM TEMPERATURE STABILITY IN MATRIX WATER QC's Theoretical Nom inal Concentrations (ng/mL) 25.0 200 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 23.8 23.1 23.8 22.9 23.0 22.9 189 191 193 193 194 194 Mean Std. Dev. n %RSD % Bias 23.3 0.432 6 1.9% -6.8% 192 1.97 6 1.0% -4.0% E D T A R A T P L A S M A Q C 's Theoretical Nom inal Concentrations (ng/mL) 34.2* 192* Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 35.1 34.2 34.0 33.2 33.6 33.5 199 200 197 195 194 188 Mean Std. Dev. n %RSD % Bias 33.9 0.674 6 2.0% -0.9% 196 4.32 6 2.2% 2.1% * = Mean concentration of six control Plasma QC's analyzed in the same analytical batch. 060C52 Primedica-Worcester Project number: PA BX -BV A 418-018-.PAG E 1-30 Page 29 TABLE 7 SU M M A R Y OF QC ST A N D A R D FREEZE-THAW (3 CYCLES) STA BILITY IN M ATRIX Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean Std. D ev n %RSD % Bias W A T E R Q C 's Theoretical Nominal Concentrations (ng/mL) 25.0 200 24.3 23.6 23.2 23.8 23.0 23.3 194 194 194 194 196 192 23.5 0.472 6 2.0% -6.0% 194 1.26 6 0.6% -3.0% E D T A R A T P L A S M A Q C 's Theoretical Nominal Concentrations (ng/mL) 34.2* 192* Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 33.5 34.4 32.9 33.6 32.3 32.4 194 200 197 193 190 184 Mean Std. Dev n %RSD % Bias 33.2 0.804 6 2.4% -2.9% 193 5.59 6 2.9% 0.5% * = Mean concentration of six control Plasma QC's analyzed in the same analytical batch. OOOC53 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-31 Page 30 TABLE 8 SUMMARY OF INTERPOLATED MEVALONIC ACID LACTONE DILUTION QC CONCENTRATIONS FOR DILUTION VALIDATION Theoretical Nom inal Concentration (ng/mL) 200 * Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 40.4 39.5 40.7 38.1 38.9 37.9 Mean Std. D ev n %RSD % Bias 39.3 1.16 6 3.0% -1.8% * = Dilution QC's undiluted concentration was 200 ng/mL (1:5 dilution performed prior to extraction and analysis) 000654 Primedica-Worcester Project number: PA B X -B V A 418-018:PAGE 1-32 Page 31 TABLE 9 6-DAY ROOM TEMPERATURE FINAL EXTRACT STABILITY BACK-CALCULATED CALIBRATION STANDARD CONCENTRATIONS Mean n % Bias 10.0 9.87 10.2 10.0 2 0.4% Theoretical Nom inal Concentrations (ng/mL) 20.0 30.0 50,0 90.0 150 19.8 30.2 50.2 89.9 19.7 30.5 49.4 90.6 150 146 19.8 2 -1.3% 30.4 2 1.2% 49.8 2 -0.4% 90.3 2 0.3% 148 2 -1.3% 250 254 253 254 2 1.4% 0G0655 Primedica-Worcester Project number: PA B X -B V A 418-018:PAGE 1-33 Page 32 TABLE 10 6-DAY ROOM TEMPERATURE FINAL EXTRACT STABILITY QUALITY CONTROL STANDARD CONCENTRATIONS Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean Std. Dev. n %RSD % Bias WATER QC'S Theoretical Nominal Concentrations (ng/mL) 10.0 25.0 70.0 200 9.89 9.17 10.5 9.75 10.7 10.1 24.3 68.8 25.1 67.7 25.5 68.7 24.4 68.3 24.7 67.7 24.3 69.9 198 198 197 200 198 199 10.0 0.550 6 5.5% 0.0% 24.7 0.492 6 2.0% -1.2% 68.5 0.826 6 1.2% -2.1% 198 1.03 6 0.5% -1.0% EDTA RAT PLASMA QC'S Theoretical Nominal Concentrations (ng/mL) 33.1 73.9 194* Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 35.0 34.4 32.8 32.2 31.6 33.7 73.5 75.6 76.2 71.4 70.7 74.9 195 189 192 191 186 185 Mean Std. Dev. n %RSD % Bias 33.3 1.31 6 3.9% 0.6% 73.7 2.26 6 3.1% -0.3% 190 3.78 6 2.0% -2.1% * = Mean concentration obtained on the first day of validation GG065 Primedica-Worcester Project number: PA B X -B V A 418-018:PAGE 1-34 Page 33 TABLE 11 SUMMARY OF CONTROL BLANK EDTA RAT PLASMA CONCENTRATIONS Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean Std. Dev. n %RSD Inter-Assay Mean n Std. D ev. %RSD Concentrations (ng/mL) Dav 1 Pay 2 Dav 3 16.4 16.5 17.0 16.2 15.9 16.8 16.6 16.3 16.8 17.4 17.4 17.3 16.3 16.1 17.1 15.8 16.2 17.0 16.5 0.536 6 3.2% 16.4 0.529 6 3.2% 17.0 0.190 6 1.1% 16.6 18 0.506 3.0% 0G0657 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-35 Page 34 TABLE 12 SUMMARY OF UNIQUE LOTS OF EDTA RAT PLASMA CONCENTRATIONS Lot 1 Lot 2 Lot 3 Lot 4 Lot 5 Lot 6 Mean Std. Dev. n %RSD Concentrations fng/mL'l 11.0 16.2 16.8 16.1 17.6 17.0 15.8 2.41 6 15.3% 0C065S Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-36 Page 35 TABLE 13 SUMMARY OF INTERNAL STANDARD PEAK AREAS Low-QC Mid-QC High-QC Water QC's Dav 1 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 4096 4380 5310 4968 4124 6187 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 4323 4390 4522 4777 5024 6314 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 4021 5545 4246 3568 5531 5458 Mean Area 4821 Dav 2 4303 4861 5511 6253 5620 6437 4827 5502 5757 5771 5881 6632 4124 5015 5029 3950 5027 6200 5372 Dav 3 5191 5911 6016 5585 6203 3846 4928 5230 5681 5479 5960 5452 5197 5314 6189 5978 7684 6335 5677 GG0653 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-37 Page 36 TABLE 13 (Concluded) SUMMARY OF INTERNAL STANDARD PEAK AREAS Low-QC Mid-QC High-QC Plasma QC's Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Dav 1 4700 4321 3850 5084 3350 5138 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 3988 4580 4925 4477 5132 4988 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 6540 5208 5002 6790 5671 3508 lean Area 4847 Dav 2 4503 5363 6736 5184 6350 5493 4091 5157 6063 3722 5601 6044 4445 5474 6088 4231 7133 5471 5397 DayJ 6100 6151 6067 5909 5159 6755 5413 6040 6238 6216 6049 5746 5985 5811 5615 6084 5879 5467 5927 Comparison of Plasma OC vs Water OC Mean Areas % Difference 0.5% 0.5% 4.4% OG0SS Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-38 Page 37 TABLE 14 SUMMARY OF PLASMA QC STANDARD CONCENTRATIONS WITH INTER- AND INTRA-DAY SUMMARY STATISTICS Dav 1 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean n % Bias % Difference vs Water QC (Table 1) Dav 2 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean n % Bias % Difference vs Water QC (Table 1) Theoretical Nominal Concentrations (ng/mL) 25.0 70.0 200 34.5 75.7 33.4 74.3 32.4 75.3 33.0 71.8 32.2 72.7 33.2 73.3 198 191 192 199 193 190 33.1 6 32.4% 73.9 6 5.6% 194 6 -3.0% -2.0% Theoretical Nominal Concentrations (ng/mL) 25.0 70.0 200 34.3 76.4 33.2 75.9 33.0 76.9 33.4 75.3 34.8 78.3 35.1 78.4 198 194 202 194 194 199 34.0 6 36.0% 76.9 6 9.9% 197 6 -1.5% 0.5% CC0C61 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-39 Page 38 TABLE 14 (Concluded) SUMMARY OF PLASMA QC STANDARD CONCENTRATIONS WITH INTER- AND INTRA-DAY SUMMARY STATISTICS Dav 3 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 Mean n % Bias % Difference vs Water QC (Table 1) Theoretical Nominal Concentrations (ng/mL) 25.0 70,0 200 34.9 75.5 198 34.5 76.8 197 35.1 77.1 199 34.2 77.9 195 34.7 76.0 202 35.7 74.7 195 34.9 6 39.6% 76.3 6 9.0% 198 6 -1.0% 0.5% Intra-assay variance estimate Inter-assay variance estimate 2.3% 2.4% 1.8% 2.0% 1.7% 0.8% Inter-Assay Mean n % Bias 34.0 75.7 196 18 18 18 36.0% 8.1% -2.0% ocoes2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-40 Page 39 APPENDIX A LABORATORY METHOD 0S0C S3 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-41 Page 40 P R IM E D IC A C O R P O R A T IO N W o rcester, M assachusetts Lab o rato ry M ethod F o r A nalysis of Mevalonic A cid in R a l P lasm a B y G C /M S D L M Num ber MVALOO R evision N u m b e r 00 Effective D ale N o vem b er 1, 2000 Page J o f 12 D ale: Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-42 Page 41 t h i ; a n a l y s is m e v a l o n i c a c i d i n r a t p l a s m a B Y C .C /M SD L M I N um ber: M V a L(HI I RKcevissiion N u m b e r: 00 E f l m i v f [>Rlc: N ovem ber I . 2000 Page 2 o f 12 1 Purpose T h e purpose o f this L a b o ra to ry M e th o d ( L M ) is to describe procedures to a c c u ra te ly d e te rm in e the concentrations o f m evaiontc acid in rat plasma samples b y G C /M S D . 2 Scope T h e procedures provided in this L M are applicable fo r the quantification o f m ev a lo n ic acid in ral plasma F.D T A . D uring the extraction m evalonic acid ( M V A ) is converted to m evalonic acid lactone ( M V L ) . T h e v a lid a te d c a lib ra tio n ran ge is 10.0 n g fm L to 2 5 0 n g /m l. o f m e v a lo n ic acid lactone in ral plasm a E D T A . T h e results can be converted to the unns o f p m o l/m l. o f m evalo nic acid in ral plasma E D T A using the conversion described in the calculation section below . 3 D fin irio n s / A b b r e v ia i ions GC: MSD. IS: MVA: M VL: M V L-d , OC: G: Gas Chrom atography M ass Selective Detector Internal Standard M evalonic Acid M e va lo n ic A cid fa c to n e or D L -M c va lo n tc A cid Lactone or M cvalonolactonc D L-M evalonolactone-4,4.5.5-d, Q u ality Control G ravitatio n al Constant 4 Materials 4.1 Chem icals D L -M e v a lo n ic acid lactone, Sigma C hem ical C o., approxim ately 9 7 % grade, or equivalent D L -M e v a lo iio la c to n e -4 ,4 ,5 ,5 -d 4.C a m b rid g e Isoto pe Lab o ra to rie s , In c ., 9 8 % grade, or equivalent Ethyl acetate, J.T. B ak e r, H P L C grade or equivalent M e th y le n e c h lo rid e , J.T'. B a k e r. H P L C grade or equivalen t 2-Propanol (isopropyl alcohol). E M Science, H P L C grade or equivalent Form ic acid, J.T. B aker, reagent grade o r equivalent C hlo ro form . J.T. B aker, H P L C grade or equivalent Sodium sulfate. A ld ric h , reagent grade, o r equivalent D eionized water. M illip o rc M illi-Q w aler, or equivalent H eliu m , Northeast A irgas, Inc., ultra h ig h p u rity or equivalent CG04o Primedia-Worcester Project number: PABX-BVA 418-018:PAGE 1-43 Page 42 T H E AN A LY S IS M E V A L O N IC A CU> IN R A T P L A S M A B Y GC.7MSD L M Number: M V A L IH I Revision N um ber: 00 E ffective D ate: N ovem ber I . 2(100 P i f r 3 of 12 lsobutane. M athcson Gas Products, research grade or equivalerti Hexanes. J .T . B aker, H P L C grade or equivalent Pooled rat blank plasma E D T A 4.2 Consum ables D is p o s a b le 15 n iL conical b o n o m p o ly p ro p y le n e tesi tubes Pipetm an and tips fcppendorf repeater and tips D isposable polyethylene transfer pipettes or equivalent Glass c rim p lo p autosum plcr vials w ith flal bottom inserts or equivalent Polypropylene vials w ith o-rutg sealed caps or equivalent 4.3 E quipm ent V W K M u lti-T u b e V o rte x c ro r equivalent Lab Industries Pipe! Dispenser or equivalent Beckm an G S -6 K R centrifuge or equivalent 7.ym ark T u rb o vap L V Evaporator or equivalent A N D E R -1 8 2 A analytical balance or equivalent J A W G C colum n, D B -2 1 0 . 3 0 m , 0.25 m m ID , 0.25 pm film thickness or equivalent H ew le tt Packard 7 68 3 injector or equivalent H ew lett Packard 6 890 gas chrom atograph or equivalent H ew le tt Packard 5973 mass selective detector or equivalent H e w le tt P ackard C h e m S ta tio n ( G 1 7 0 1 B A , R ev is io n B .01 DO) or equivalen t M ic ro s o ft Excel y7 version S R -2 or equivalent Note: E qu ip m en t, reagents, or consum ables can be substituted provided that equivalent assay p e rfo rm a n c e is dem onstrated. 5 P rocedure This m ethod has been validated fo r the analysis o f m evalonic acid (as m evalonic acid lactone) in rat plasma E D T A by G C 7 M S D using 100 p L sam ple volum es. T h e validated concentration range is 1 0.0 n g /m L to 2 5 0 n g /m L o f m e v a lo n ic acid lactone in rat plasm a E D T A . T h e c o n version o f th e results to p m o l/m L o f m ev a lo n ic acid in rat plasm a E D T A is described in the calc u la tio n s section b e lo w . N o te that the c alib ra tio n c u rv e and q u a lity control standards arc all prepared in w a te r, not plasm a. M e v a lo n ic acid is a n a tu ra lly occu rrin g com pou nd in plasm a and concentrations w ill v a ry w ith diet. C o n c e n tratio n s found in po oled lots o f b la n k rat plasm a E D T A du rin g the m eth o d valid a tio n w e re a p p ro x im a te ly 17 n g /m L . 'Phis background c oncentration le v e l w ould interfere w ith the quantitailon, especially ai the low er calibration level. This m ethod has been validated to use w ater m atrix calibration curves and qu ality control samples. T h e validatio n dem onstrated that the m ev a lo n ic a cid lactone calculated results fo r the w ater m atrix is e q u iv a le n t to rat plasm a E D T A . GG0GG* Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-44 Page 43 THE. A NA LYSIS M E V A L O N IC A C ID IN R A T P L A S M A BY G C /M SD L M N um ber: M V ALIMI Revision Num ber: 00 Effective H a lt: N uveinber 1, 200(1 Page 4 o l 12 5 .1 Preparation o f Reagents N ote: O th er volum es than those specified m ay lie prepared using the sam e proportions. 5.1.1 M e th y le n e C h lo rid c :2-P ro p an o l, (9 0 :1 0 , v:v) Solution C o m b in e 9 0 0 m l . o f m e th y le n e chlo ride w ith 10 0 n tL o f 2 -p ro p a n o l and m ix . Store at room te m p era tu re . T h e e xp iration date for this s olu tion is o n e w e e k after preparation. 5.2 Preparation o f Internal Standard Solutions C oncentration calculations are based on assuming that the standard reference m aterials are 10 0 % pure unless no ted o th e rw is e . T h e fo llo w in g standard p reparation schem e is a suggested approach. A pprop riate m odifications to reach the targeted nom inal concentrations are acceptable and w ill be documented in the study notebook. A ll standard solutions w ill be stored at < -2 0 C unless noted otherwise. Th e exp iration dates fo r these solutions arc six m onths a fte r preparation unless noted otherw ise. O n g o in g stab ility studies m ay change the exp iratio n dales. 5.2.1 Internal Standard Stock (IS A ) M V L - d , is a liq u id at ro o m tem p erature and is purchased in v ia ls c o n ta in in g a p p ro x im a te ly 10 0 m g . W e ig h the v ia l and M V L - d 4. T ra n s fe r th e M V L - d , to a 50 m L volu m etric flask. W e ig h the empty via l. Subtract the w eights to obtain the am ount o f M V L - d , in the volu m etric flask. B rin g to v o lu m e w ith M il li- Q w ater and m ix . T h e n o m in a l M V L - d 4 IS concentration o f this solution is 2 0 0 0 p g /m L . 5.2.2 Internal Standard S piking Solution Transfer 2 0 .0 p L o f in tern a l standard stock ( IS A ) into a 1 00 m L v o lu m e tric flask. D ilu te to volum e w ith M illi-Q w ater and m ix. T h e nom inal M V L -d , concentration o f this s o lu tio n is 4 0 0 n g /m L . 5.3 Preparation o f C alib ratio n S tandard Slock Solutions T h e fo llo w in g standard p re p a ra tio n schem e is a suggested approach. A p p ro p ria te m odifications to reach the targeted nom inal concentrations arc acceptable and w ill be documented in the project no teb ook. Fo r exam ple, i f the targeted no m in al concentration is not ach ieved w h e n p re p a rin g th e a n a ly tic a l c alib ra tio n stocks, the v o lu m e o f the slock solutions used m subsequent d ilu tio n s can be m odified so that the no m inal calibration standard spiking solution concentrations are achieved. Standard reference m aterials are assumed to be 100% pure. Standard w eights listed are not corrected fo r pu rity. A ll standard solutions w ill be stored at -2 0 C unless noted oth erw ise. O n g o in g s ta b ility studies arc establishing th e e x p ira tio n dales o f these solutions. 5.3.1 M V L P rim ary Slock ( A ) 0C0667 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-45 Page 44 TIJ A N A LY S IS M E V A L O N IC A C ID IN H A T P LA S M A BY G C /M S l) t.M Number: M V A 1.00 K r v i s i u D N u m b e r: 00 E ffe ciiv c D o le : N ovem ber 1, 2000 Page 5 of 12 N o te ih al M V ] is hyd ro sco p ic. A large am ount o f M V L is used and it is w e ig h e d out q u ic k ly to m in im ize the effects o f absorbing m oisture out o f the air. Siore the a n a ly tic a l reference standard under an inert gas such as nitrogen. 5 .3 .2 5 .3 .3 5 .3 .4 W e ig h out a p p ro x im a te ly 100 m g o f M V l , and transfer tt in to a 100 m L volu m etric flask. B rin g to v olu m e w ith ethyl acetate and m ix . T h e nom inal M V L conc e n tra tio n o f this solution is 1 000 p g /m L . Secondary Stock (B ) T ra n s fer 3.75 m L o f the P rim ary Stock (A ) in to a 2 5 .0 m L volu m etric flask. B rin g to v o lu m e w ith eth yl acetate and m ix . T h e n o m in a l M V L concentration o f this solu tio n is 150 p g /m L . Preparation o f C alibration Standard Spiking (C S S ) Solutions T h e fo llo w in g standard p re paration schem e is a suggested approach. A p p ro p ria te m odifications to reach the targeted nom inal concentrations are acceptable and w ill be docum ented in the study notebook. A ll standard solutions w ill be stored at < 2 0 C unless noted otherwise. O n g o in g stability studies are establishing the exp iratio n dates o f these solutions. P rep are C S S s olu tion s in 2 5 .0 m L v o lu m e tric flasks as sh o w n In the ta b le belo w . A ll solutions are brou ght to v o lu m e w ith ethyl acetate. F in a l volum es o f these c a lib ra tio n standard spikin g s olu tion s can be adjusted as lo ng as p ro p o rtio n a lity is m a in ta in e d and th e ir preparation is p ro p erly d o cu m e n te d in the study n o teb ook. S p ik in g S o lu tio n ID CSS-1 CSS-2 CSS-3 CSS^l CSS-5 C S S -6 C S S -7 C a lib ra tio n S ta n d a rd S p ik in g .Solution P re p a ra tio n Stock S olution Stock S olution Stock S olution S pikin g S olution V o lu m e C oncentration F in al V olum e ID <pL) (p g /m l.) (n iL ) Secondary Stock B 16S 150 2 5 .0 Secondary Slock B 337 150 2 5 .0 Secondary Stock B 505 150 2 5 .0 Secondary Slock B 842 150 2 5 .0 Prim ary Stock A 227 1000 2 5 .0 Prim ary Stock A 379 1000 25.0 Prim ary Stock A 631 100O 2 5 .0 S piking S olutio n C oncentration (n g /m l.) 1010 2020 3030 5050 9090 15150 25250 5.3.5 O nce prepared, aliquot the C alibration Spiking Solutions (suggested approxim ately 0 .6 m l volum es) into plastic vials w ith tv rin g sealed caps and store them at S -2 0 " C u n til the lim e o f analysis. 5.4 Preparation o f C alibration Standards (S T D ) in W ater 0G06S& Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-46 Page 45 1 H t A N A L V M S M E V A L O N IC A C ID IN R A T P L A S M A BY C.CTMSO l.M Number: M V A LD 0 RrvLsion N u m b e r: 00 Effective D ale: N ovem ber I. 2000 Page 6 of 12 ------------------ ^ C alib ratio n S ta n d a rd ID STD 1 STD 2 STD 3 STD 4 STD 5 STD 0 STD 7 Prepare calib ratio n standards on the day o f analysis by c o m b in in g 2 .5 0 m L o f M illi-Q w ater, in d iv id u a lly , w ilh 0 .0 2 5 0 mL- o f the respective C a lib ra tio n S p ikin g Solution (C S S ) fo llo w e d b y m ix in g . T h e fin a l concentration in w a te r is listed in th e table belo w . W ater V olum e (m l.) 2 .5 0 2 .5 0 2 .5 0 2 .5 0 2 .5 0 2 .5 0 2 .5 0 C a lib ra tio n S tandard P rep aratio n S p ikin g S olution S p ik in g S p ikin g S olution V o lu m e A tUletl S u lu tiu n C o n centration <m L) ID tn g /m l.) 0 .0 2 5 0 CSS-1 1010 0 .0 2 5 0 CSS-2 2020 0 .0 2 5 0 C S S -3 3030 0 .0 2 5 0 C S S -4 5050 0 .0 2 5 0 CSS-5 9090 0 .0 2 5 0 C S S -b 15150 0 .0 2 5 0 C S S -7 25250 F in al N om inal Plasm a C one. (n g /m L ) 10.0 20.0 30.0 50.0 90.0 150 250 5.5 Preparation o f Q u ality C o n tro l (Q C ) Stock Solutions T h e fo llo w in g standard p re paration scheme is a suggested a pproach. A p p ro p ria te m o d ificatio n s to reach the targeted nom inal concentraiions are acceptable and w ill be docum ented in the project notebook. Standard reference m aterials are assumed to be 1 0 0 % pure. Standard w eights listed are not corrected fo r pu rity. A ll standard solutions w ill be stored at < - 2 0 "C unless noted otherwise. O n g o in g stab ility studies are establishing ih e exp iratio n dales o f these solutions. 5.5.1 Prim ary Q C Stock ( A A ) N o te that M V L is hydroscopic. A large am ount o f M V 1 . is used and it is w e ig h e d out q u ic k ly to m in im ize the effects o f absorbing m oisture out o f the air. Store the a n a ly tic a l refe re n c e standard under an inert gas such us n itro g e n . 5 .5 .2 W e ig h out a p p ro x im a te ly 1 00 m g o f M V L and tran sfe r it in io a 1 0 0 m l, volu m etric flask. B ring to volum e w ith ethyl acetate and m ix . T h e nom inal M V L c o n c e n tra tio n o f this solution is 10 0 0 p g 'm L . Secondary Q C Stock (B B ) ira n s fe r 5 .00 m l. o f the Prim ary Q C S lo ck ( A A ) in to a 2 5 .0 m l. volum etric flask. B rin g to v o lu m e w ith ethyl acetate and m ix. T h e no m inal M V L concentration o f this soiulion is 2 00 p g /m L . 000669 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-47 Page 46 I H t A N A L Y S IS M E V A L O N IC A CII IN R A T P LA S M A BY C C /M S D I.M Number: M V A I.0 0 Revision N um ber: 00 Effrctive Date: Novem ber i. 2000 Page 7 of 12 5.6 JTepuralion o f Q u ality C ontrol S piking Solutions (O C S S ) T h e fo llo w in g standard preparation scheme is a suggested approach. A pprop riate m o d ificatio n s to reach the targeted nom inal concentrations are acceptable and w ill be d o cu m e n te d in th e study n o teb o o k . A ll standard solutions w i ll be stored at S -2 0 ? C unless noted otherw ise. O n going s tab ility studies are establishing the exp iration dates o f these solutions. Prepare O C S S solutions in 2 5 . ra L volu m e tric flasks as shown in the tab le b e lo w . A ll solu tion s are brought to v o lu m e w ith ethyl acetate. F in a l v olu ntes o f these q u a lity control sp ikin g solutions can be adjusted as long as p ro p o rtio n ality is m aintained and their p re p a ra tio n is p ro p e rly do cu m ented in th e s tudy n o teb o o k . S piking S o lu tio n JO Q C SS-LO W Q C SS-M ED Q C S S -I1IG H Q C S tandard S piking S olution P rep a ra tio n Stock Stock S p ik in g Stock S olution S olution S o lu tio n S o lu tio n 11) V olum e Cone. Final V olum e Used Secondary Q C Slock BB (p l-) 316 (p g /m L ) 200 (m l,) 25.0 Secondary Q C Stock B B 884 200 25.0 P rim ary Q C Stock A A 505 1000 25.0 S p ik in g S olution Cone. (n g /n iL ) 2525 7070 20200 5.7 Preparation o f Q u ality C o n tro l (Q C ) Standards in W ater C o m b in e 100 raL o f M illi-Q w ater, in d ivid u ally, w ith 1.00 m L o f each Q u ality C ontrol S p ik in g S o lu tio n ( Q C S S ) as show n on th e ta b le b e lo w . Q u ality C ontrol Standard ID l.o w Q C M id Q C H igh Q C Q u ality C o n tro l S tan d ard P rep aratio n Stock Stock Solution S tock S olution F in a l .Suint ion C oncentration S pikin g V o lu m e V olum e. ID (n g /n iL ) (m l.) (m L ) Q C S S -LO W 2525 1.00 100 Q C S S -M E D 7070 1.00 100 Q C SS-H 1G H 20200 1.00 1 100 Q C Standard C oncentration (n g /m L ) 2 5 .0 7 0 .0 200 5 .7 .1 O n c e prepared, aliq u o t the O C standards (suggested a p p ro x im a te ly 1.2 m L volum es) into plastic vials w ith o -n n g scaled caps and store them at l;-2 0 C until the tim e o f analysis. C067# Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-48 Page 47 T H E A N A LY S IS M E V A L O N IC ALTO IN R A T P LA S M A B Y G C /M S U L M Num ber: MVAl.on Revision N um ber: 00 Effective D ate: N ovem ber 1, 1000 Page o f 12 5.8 Sample Extraction 5.8.1 Transfer 100 p L o f each standard (total o f 14), m ethod blanks (in duplicate), control blanks (in d u p lic a te ), O u a lity C o n tro l Standards (du p lica te s at three concentration levels), and study samples into in dividual 15 m l. conical bottom polypropylene test tubes. 5.8.2 A dd 50.0 p L o f M illi-Q w ater to each m ethod blank. 5.8.3 Add 5 0 .0 p L o f the Interna! Standard S piking Solution to each tube except for the method blanks. 5.8.4 V o rte x the tubes fo r a m in im u m o f 5 seconds. 5.8.5 A dd 0.100 m L o f form ic acid to each lube and vortex for a m inim um o f 5 seconds. 5.8.(> A llo w tubes to stl on th e bench for a p p ro x im a te ly 3 0 m inu tes to a llo w tim e for the conversion o f M V A to M V L . 5 .8 .7 A dd 0 .8 0 0 m L o f M illi- Q w ater to each lube and vortex for a m inim um o f 5 seconds. 5 .8 .8 Add 0 .500 m l, o f chloroform to each tube ami vortex for a m inim um o f 5 seconds. Note: a pipe! dispenser m ay be used lo add this reagent. 5 .8 .9 C e n trifu g e th e lu b e s a p p ro x im a te ly 5 m inu tes at a p p ro x im a te ly 3 5 0 0 rp m (a p p ro x im a te ly 2 5 0 0 G ). D is tin c t layers should fo rm in each sam ple tube. C en trifuge again any sam ple w h ich does not have distinct layers. 5 .8 .1 0 T ra n s fe r the lo p (aq u e o u s) la y e r in to n e w in d iv id u a l 15 m L con ica l bottom polypropylene test tubes and discard the choroform layer. 5.8.11 Saturate the aqueous layer w ith sodium sulfate. (N o te: approxim ately 0 .15 grams o f s odium sulfate is p le n ty to saturate th e aqueous la y e r. A m easuring spoon m ay be used lo add the sodium sulfate because the exact w eight is not critical.) 5 .8 .1 2 A dd 1.0 m L o f m ethylene chloride.2-propanol solution to each tube containing the aqueous layer and vortex fo r a m in im u m o f 5 seconds. Note: a pipe! dispenser m ay be used to add this reagent. 5 .8 .1 3 C en trifu g e the tubes a p p ro x im a te ly 5 m inutes at a p p ro x im a te ly 350 0 rpm (approxim ately 2 50 0 G ). 0671 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-49 Page 48 T H E A N A LY S IS M E V A L O N IC A C ID IN R A T P LA S M A BY GC7MSD L M Number: M V A ld U Revision N um ber: 0 Effective Dale: November 1,2000 Page 9 o f 12 5 .8 .1 4 T ransfer ilie b o tto m (o r g a n ic ) liiyer in to new 15 m l. conical bottom polypropylene lest tubes. 5.8.15 Repeat the previous three steps. 5 .8 .1 6 F.vapurate the o rg a n ic la y e r to dryness in a 'J u rbn vap at 4 0 5 " C at a p p ro x im a te ly lO psi. 5.8.17 A dd 50 uL o f the ethyl acetate to each tube and vortex for a m in im u m o f 5 seconds. 5 .8 .1 8 T ransfer into crim p to p autosam p ler vials w ith flat bottom inserts. Extracts arc stable in aulosam pler vials stored at room temperature for up to 6 days. N o te: I f the extracts evaporate to dryness w h ile w aiting for analysis they m a y be reconstituted w ith m ore ethyl acetate before injecting. 5.9 A nalytical Conditions G C /M S D analysis is p e rfo rm e d as fo llo w s : 5.9.1 Oven Colum n: In itial Tem perature: Initial Tim e: Oven Ramp: R unTim e: J & W , D B -2 1 0 , 30 m, 0.25 m m ID , 0.25 pm film thickness 50C 1.50 m inutes R ate Final Tem p. Final Tim e |C /m u i.) (C ) (m in.-) 3 0 .0 160 3 .5 0 5 0 .0 230 3 .5 0 a p p ro x im a te ly 14 m inutes 5.9.2 Inlet Injection V olum e: Rinse Solvent: Mode. Tem perature: Purge Flow: Purge Tim e: Gas Type: In itial Flow: Flow Ramp: l pL Hexanes S p litle s s 2 40 "C 50.0 m U m in u tc 0 .7 5 m inutes F le iiu m 3.5 m lV m inutc Constant F lo w M ode 000672 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-50 Page 49 T H E A N A L Y S IS m e v a l o n i c a c i d i n R A T p l a s m a B Y C O M SI) l.M Number: M VAI.OO Revision N um ber: 00 Etlecnve Dato: Novem ber 1, 20IMI Page 10 o f 12 5.9.3 Mass Selective D etector Source: Source M ode: C l Gas: F lo w C ontrol Setting: Acquisition M ode: Solvent Delay: 5.9 4 M S Quad: M S Source: M S D Transfer Line: Ions M on itored: Chem ical Ionization (C l) Positive Isobutane 20 Selective Ion M o n ito rin g ( S IM ) 6 .0 m inu tes (m a y be adju sted to be as long as possible w ithou t elim in atin g the first peak o f interesi) 106-C 250C 2 5 0 gC 5 .1 0 5 .1 ) 5.9.4.1 G ro u p 1: Start tim e a p p ro x im a te ly 6 .0 0 ' Compound N om inal Ion D w ell Retention T i m e *1 T une1 Nam e M VL issi* ) 13) (m sec.) 100 (m in.) 7.2 M V L -d , 135 100 7.2 1 = D w e ll tim e s m ay b e m o d ifie d to o p tim iz e p e rfo rm a n c e . - = A p p ro xim ate retention tim e (R T ) for a new colum n. A ctual R T m ay v a r y as the c o lu m n is shortened d u rin g m ain te n a n c e . T h e R T is confirm ed using the isotopically labeled internal standard. ' = Start tim e m ay be adjusted. A nalytical Run Sequence and Com position Each analytical run w ill be com prised o f bracketing water calibration curves (7 points) each w ith a w ater m ethod b lan k sam ple ( water w ith neither analyte n o r internal standard spiked) and a w ater control blan k (w ater w ith o n ly internal standard included). Duplicate w ater Q C standards at each o f the three concentrations w ill be interspersed between the calibration curves. Plasm a D ilu tio n Q C ' s w ill lie analyzed w ithin analytical batches m w h ich plasm a samples are analyzed at a dilution. T h e d ilution o f plasma samples and plasma Q C 's w ill be perform ed using w ater as th e diluent C a lc u la tio n s 5.11.1 C hrom atogram s w ill be a u to m atically integrated and visually inspected fo r an acceptable integration using H P Chcm.Siation (o r equivalent). M an u al integrations w ill be perform ed when necessary. 000673 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-51 Page 50 T H E A N A LY S IS M E V A L O N IC A C ID IN R A T PLA SM A BY G C /M S D l,M Number: MVALOO Revision N um ber: 00 ~ E f f e c t i v e Dale: November 1,2000 PRe I I of 12 5 . 11.2 C o m p u te the l / X : -w e ig h te d least-squares linear regression re la tin g the peak area ratios (relativ e to internal standard) o f the calibration standards to their respective n o m inal concentration (n g /m L o f M V L in w ater) using valid ated Excel spreadsheets (or equivalent). 5 .1 1 .3 U sing the peak area ratios o f the analyte (relative to the in ternal standards) in the samples and the approp riate regression equation constants, d eterm in e the concentration in n g 'm L o f M V L in w ater or plasm a for alt standards and samples using validated E x c e l spreadsheets (o r equivalent). C orrect for dilutions if appropriate. 5 . 1 1.4 I f requested, the units o f n g /m L o f M V L at w ater or plasma m ay be converted to p tn o l/m L o f M V A in W3tta or plasma by m u ltip lyin g by a un it conversion value (1 0 0 0 pm oi g/m o l ng) and dividing by the m olecular w eig ht o f M V L (1 3 0 .1 4 g /m o l). U elo w is an exam ple showing the conversion o f a sam ple concentration o f 25 n g /m L o f M V L in plasma. X2 5 .0 t* g lO OOpm ol g X In to ! = m l. w t nyt 130.14 g 192pm ol ntL 5.12 Acceptance C riteria 5.12.1 C alibration Standards 5 .1 2 .1 .1 T h e low er lim it o f quantitation (L L O Q ) standard's back-calculated concentration must be w ith in 2 0 % o f theoretical n o m in a l concentration. 5 .1 2 .1 .2 A ll other standards' back-calculated concentration must be w ithin 1 5 % o f their nom inal theoretical concentrations. 5 .1 2 .1 .3 A m inim u m o f 7 5 % o f the calibration standards m ust m eet the backcalculated cuncenlralion criteria. (N ote: this lim it w ill be rounded dow n when using 14 calibration standards. T h u s, a m ax im u m o f 4 standards m ay be rejected when analyzing a total o f 14 calibration standards in a batch.) 5 .1 2 .1 .4 A t least one standard at the L L O Q and at the u p p e r lim it o f quantitation (U l.O Q ) must meet the back-calculated concentration criteria. OC0674 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-52 Page 51 T H E A NA LYSIS M E V A L O N IC A C ID IN R A T P LA S M A BY G C fM SD I.M N um ber: M V A I.00 R evision N u m b e r: 01) K fleciive D ate: N o v e m b e r 1, 2000 Page 12 o f 12 5 .1 2 .2 Q u a lity C ontrol (Q C ) Standards 5 . 12 .2 .1 T h e lowest level Q C 's calculated concentration must be w ith in 2 0 % o f the nom inal theoretical concentration. 5 .1 2 .2 .2 A ll other O C 's calculated concentration must be w ith in 1 5 % o f their nom inal theoretical concentration 5.1 2 .2 .3 A m in im u m o f h 6% o f all Q C 's must meet the calculated concentration criteria. 5 .1 2 .2 .4 A t least o n e Q C a t t a c h concentration level m ust m eet the c a lc u la te d concentration criteria. 5 .1 2 .3 B lanks and Controls A c o n tro l blank should h a v e a response (peak area ra tio ) less than o r e qual to 3 3 % o f the lowest accepted L L O Q calibration standard 5.13 T h e control blanks (in w a te r) should have a response (peak area ratio ) < 3 3 % o f the lowest accepted l.l.O Q calibration standard. Data Reporting 5 . 1 3 . 1 S a m p le concentrations less than the low est calib ratio n standard w i ll be fla g g ed as B elo w the Quantitation L im it (B Q L ). 5 .1 3 .2 S am p le concentrations greater than the highest calibration standard w ill be appropriately diluted w ith M illi-Q water, re-cxiractcd. and re-analyzed, provided that enough sam ple v o lu m e rem ains. I f there is in s u ffic ie n t sam ple a v a ila b le , the o rig in a l result w ill be reported as A b o v e the Q uan litn lio n 1 im it (A Q L ). 6 Revision History In itial laboratory method. G 073 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-53 Page 52 APPENDIX B REPRESENTATIVE CHROMATOGRAMS FROM VALIDATION DAY 1 GC0G7S Primedica-Worcester Project number. PABX-BVA 418-018:PAGE 1-54 Page 53 Peak Area Report Sample Name: l x Blank Water N otebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 13.52 O perator MAN Data File Name: 0621001.D Data File Path: D:\MSO_OATA\PAB\PABX-BVA\1-062-1\ Instrum ent Method Nama: PABX_BVA Instrum ent Name: MS7 Instrum ent Vial Number: 1 Compound * 1) 2) Compound Name MVl MVL-CW R e te n tio n Time iM in.i 000 0.00 tec 131 135 Peak Area M' 0 0 Peek Area Ratio #0IVA)! W " The letter "In" In this column Indicates that manual Integration was performed on that compound. Print Date and Time: 10/18/2000 12:39 Paqe 1 00*677 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-55 Page 54 Quantitation. Report (QT Reviewed) Data File : D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621001.D Vial: 1 Acq On : 17 Oct 2000 13:52 Operator: MAN Sample Misc : lx Blank Water : Inst : MS'? MS I n t e g r a t i o n Params: r t e i n t . p Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) jMjndftncft TIC. 0621001 D : 190, 180 i ito ,; , i; 160f... V ' w v W ^ > -/" ! i 1 '* \ . \ >J A - . . w s A*' fTime- 6.10 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 700 710 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 MVL Amount: Not Found Ion: 131 (0.000) r. 4 f\r* . 8.20 6.30 6J0_ 6.50 6.60 6.70 6-BO 650 7.00 7 10 7.20 7.30 7 0 7,50 7 60 7 70 7.B0 7.90 ^^4VIi-d4 Amount: Not Found Ion: 135 (0.000) !\ A, V,'V-...- / 5.20 6 .X 6.40 6.50 6 60 6.70 6.80 6.30 700 7 10 7.20 7 30 7.40 7.50 7.60 7.70 7.80 7 " 0 6 2 1 0 0 1 . D PABX BVA.M Wed O c t 18 1 2 : 4 0 : 0 3 2 0 0 0 Page 2 00067 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-56 Page 55 Peak Area Report Sample Name: 1x Std-1 Water Notebook Reference: PABX-0VA-1-O62 Date A c q u ire d : 10/17/2000 14:11 O p e ra to r MAN Data FHe Name: 0621002.D Data File Path: D:\MS0J3ATA\PAB\PABX-BVA\1 -062-1 \ Instrum ent Method Name: PABX-BVA In stru m e n t Name: MS7 Instrum ent Vial Number: 2 Compound # U 2) Compound Name MVL MVL-d4 R e te n tio n Time (Min.i 7 15 7 17 km 131 135 Peak Area M* 250 3280 Peak Area Ratio 762.2E-4 M* The letter "m" in this column indicates that manual inlet)ratton erma performed on that compound. Print Date and T im -10/18/2000 12:40 Patj*? 1 OOCV3 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-57 Page 56 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621002.D Vial: 2 17 Oct 2000 14:11 Operator: MAN lx Std-1 Water Inst : MS7 MS I n t e g r a t i o n P a r a m a : r t e i n t . p Quant Method : D:\KPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundarte 1500 TIC' 0621002 D 500 "I 1 1 ' 1 I rf "" I 1 Tin- 6 10 6.20 6.30 6.40 6.50 6.60 6.70 6 80 MVL ~ I o n : 131 (7.149) .. j Resp: 250 ................ T ''' ' I ' ' 1 --- - j-n -,r '--- r-- 'r-j-r-ri--- ;--------- i 6 90 7 00 7 10 7.20 7 30 7 40 7.50 7.60 7.70 7.80 7.80 J A m o u n t: _ 0 .0 0 " ... .. .... i A i\ ! 6 20 6 30 6.40 6.50 6.60 6.70 6 80 6.90 7 00 7.10 7.20 7.30 7.40 7.50 7.60 ... 7.70 7 80 7.90 -d4 " Ion: 135 (7.173) Amount: 0.00 !Resp: 3280 A t\ 6.20 6.30 6.40 6.50 6.60 6 70 6 80 6.90 7.00 7.10 7.20 7.X 7 40 7.50 7.60 7 70 7.B0 790 0 6 2 1 0 0 2 . D PABX B V A .M Wed O c t 18 1 2 : 4 0 : 2 2 2 0 0 0 Page 2 GCG0 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-58 Page 57 Peak Area Report Sam ple Name: 1* Std-2 Water N otebook Reference: PA8X-8VA~1'062 Date Acquired: 10/17/2000 14:30 Operator: MAN Data File Name: 0621003.D Data File Patti: D \MSD_DATA\PAB\PABX-evA\1-062-1\ Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Instrum ent Viai Number: 3 Comoound # it 2) Comoound Name MVL MVL-tM R eten tio n Tim e (Min.) 7.1 7.17 ion 131 135 Peek Area M* S67 m 3984 Peak Area Ratio 142:3E-3 M" * Tire lattar "m" in ttii column Indicata that manual integration wa* performed on mal compound. Print Date and Time: 10/18/2000 12:40 i ooocbi Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-59 Page 58 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621003.D Vial: 3 17 Oct 2000 14:30 Operator: MAN lx Std-2 Water Inst : MS7 MS Integration Pararne: rteint.p Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance !, * " 1 i !50o! nC: 0621003.0 1 j ! 1000- I I! 500! j i - - - - - - - r - I , r ' T T T i - - - - - - - - - - p . ' I- - - - - - - i - f - p - V - t - n - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - i . f r .-- ' P |Tirner 6.1.0.. 6.20 6.30 5 40 6.50 6.60 6.70 6.B0 6.90 7.00 7.10 7.20 7.30 740 7.50 MVL ___________________________________________A m o u n t :________ 0 . 0 T ...I o n : 1 3 1 .... ( 7 . 1 4 4 ) ! Reap: 5G7 7 60 0 - - - - - -- - - - - ' ~ -- 7.70 7.80 7.90 _______________ I ! ....... r - i - 'T-- ' 'I ' ' 1 1 *T ' 1 1 I ' ....................... ! r ' : . . i ' r ' . ------ r -- -P 1 ................. ...........................n .........T -- ' T - ' - ' i ! : 6.20 6.30 6.40 6.S0 6.60 6 70 6 80_...6 90 7 00 7.10 7.20 7.30 7 40 7.50 7,60 7 70 7.80 7.90 0 6 2 1 0 0 3 . D PABX BVA.M Wed O c t 18 1 2 : 4 0 : 4 1 2 0 0 0 Page 2 GC0682 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-60 Page 59 Peak Area Report Sample N am *: 1x Std-3 Water Notebook Reference: PABX-BVA-1-062 Data A cq u ire d : 10/17/2000 U 50 O perator: MAN Data File Name: 0621004.0 Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1-062-1\ Instrum ent Method Name: PABX_BVA In stru m e n t Name: MS7 Instrum ent Vial N um ber 4 Comoound # i) 2) C om oound Name MVL MVL-d4 R eten tio n Time IMin.l 7 15 7.17 Ion 131 135 Peak Area M* 901 m 4234 Peak Area Ratte 212.8E-3 M* * The la tte r " m " In th is c o lu m n in d ica ta th a t m anual in te g ra tio n ara p e rfo rm e d o n th a t com pound. Print Date and Time. 10/18/2000 12:40 Page 0G0683 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-61 Page 60 Quantitation Report <QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621004.D Vial: 4 17 Oct 2000 14:50 Operator: MAN lx Std-3 Water Inst : MS7 m s Integration Params: rteint.p Quant Method : D:\HPCHEM\l\MBTHODS\PABX_BVA.M (RTE Integrator) !w>undianc* TIC: 0621004.0 1500: 1000: Ij S0 -I ;\ (Time- 6 10 6 20 6.30 6 ^ 4 0 ^ ^ 5 0 6.60 6 70 6.80 6.90 7.00 7,10 7.20 7 30 7 40 7.50 7.60 770 7.80 7.90 MVL... _________ :______________ Amount : 0.00 : Io n :~ "l3 1 (7 .1 4 6 ) I K esp: 901 620 6 30 6 40 6.50 6.60 6 70 6.80 6 90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7,70 7 80 7.90 'L-d4 _______________ w - Ton: 135 (7T169) Amount ; 0.00 Reso: 4234 T-, , , , 6.20 6.30 6.40 6.50.....6.60 670 6.80 6.90 7 0 0 7.10 720 7.30 7.40 7.50 7.60 7.70 780 7.90 0 6 2 1 0 0 4 .D PABX_BVA.M Wed O ct 18 1 2 : 4 1 : 0 1 2 0 0 0 Page 2 GC0S34 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-62 Page 61 Peak Area Report Sam ple Name; 1* Std-4 Water N otebook Reference: PABX-BVA-1-062 Date A cquired: 10/17/2000 15:09 O perator: MAN Data Fite Name: 0621005. D Data File Path: D:NMSD_DATA\PAB\PABX-BVAV\-Q62-1\ Instrum ent Method Name: PABX_BVA instrum e n t Name: MS7 instrum ent Vial Number: 5 Compound 1) 2) Comoound Nome MVL MVL-d4 R e te n tio n Time (Min i 7.14 7 17 ion 131 135 Peek Area M' 1285 3749 Peek Area Ratio 342.8E-3 h r * The tetter "m" in itti column indtcaie* that manuel integration waa performed on that compound. Print Date and Time 10/18/2000 12:41 Pay* I 010G8 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-63 Page 62 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621005.D Vial: 5 17 Oct 2000 15:09 Operator: MAN lx Std-4 Water Inst : MS7 MS Integration Params: rteint.p Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTB Integrator) (Abundance ;I 1500 TIC: 0621005.D A, r ! ; loooi i I 6.10 6.20 6.30 6.40 6.50 6.60 6.70 680 690 7<Xl' T IP 7.20 7 .3 }' ' t V 7.SO 7.60 7 70 7 80 7.90 MVL ____________________Amount : 0.00 i! Ion: 131 (7.143).. Reap: 1285 6.20 6.30 6 40 6.S0 660 6.70 6.80 6.90 7.00 7.10 7 20 7 30 7 40 7S0 7 60 7.70 7.80 7.90 L-d4 9 F Ion: 135 (7.171) Amount : 0.00 Resp: 3749 :AI II \ --t i . ! -.'i-r-i-i -- r ....... : " - " : I 1-- f~ r6.20 6.30 6 40 6.50 S.60 6 70 6.80 690 7.00 7.10 7-20 7 30 7 40 7.50 7.60 7.70 7.80 7.90 0 6 2 1 0 0 5 .D PABXJSVA.M Wed O c t 18 1 2 : 4 1 : 2 0 2000 Page 2 0C 0 6 8 * Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-64 Page 63 Peak Area Report Sam ple Name: 1 Sid-5 Water Notebook Reference: PABX-8VA-1-062 Dale A cquired: 10/17/2000 15:29 Operator: MAN Data File Name: 0621006 D Data File Path: D:\M5D_OATAIPAB\PABX-BVAU-062-1\ Instrum ent Method Name: PABX_BVA Instrument Name: MS7 Instrum ent Vial Number: 6 Comoound * 1) 2) Comoound Name MVL M V U tt R e te n tio n Time <Mbv) 7.14 7.17 ion 131 135 Peak Area M* 2208 3693 Peak Area Ratio 597.9E-3 W The letter "m" in this column indicales that manual Integration was pertormadon that compound. PnntOats and Time: 10/18/200012.41 FAyn 1 0C0687 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-65 Page 64 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BVA\l-062-1N0621006.D Vial: 6 17 Oct-2000 15:29 Operator: MAN lx Std-5 Water Inst : MS7 MS Integration Params: rteint.p Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC: 0621006.0 1500; / 1000- / ! 500- --! -1 -- >U .. T r' 1 ' r ' I i I I'I---r-rj-i -: r ,r r r i-n I --r-r-- 7-- Tr*-> 6 10 6.20 6.30 6.40 6.50 660 6.70 ... 6.80 6.90 7.00 7.10 7.20 T.30 7 40 7.50 7.60 770 7 60 7 90 MVL Am ount : 0.00 Ion:.131 (7.144)' Reap: 2208 /i .. 6.20 6 30 6.40 6S0 6.60 6.70 6 80 6.90 7.00 7.10 7.20 7.30 7 <0 7.50 7.60 770 780 7,90 L-d4 ^F~'lIoin: 135 (7.172) Amount: 0.00 j Reap: 3693 A 1I T1 ' I' 1 . . r-- 6.20 6.30 6.40 650 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7.30 7 40 7.50 7.60 7 70 7B0 0 6 2 1 0 0 6 .D PABX_BVA.M Wed O c t 10 1 2 : 4 1 : 3 9 2 0 0 0 Page 2 000688 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-66 Page 65 Peak Area Report Sampkr Nam: lx Std-6 Water N otebook Reference: PABX-BVA-1-062 Date A cq u ire d : 10/17/2000 15 48 Operator: MAN Data FHe Name: 0621007.0 Data FHe Path: D:1MSD_DATA\PAB\PABX-BVA\1-062-H Instrument Method Name: PABX_BVA In stru m e n t Name: MS7 Instrum ent Vial Number: 7 Compound # 1) 2) Com pound Name MVt MVL-<14 R e te n tio n Time fMin.1 7.14 7.17 Ion 131 135 Peak A le a M* 4054 m 4062 Peak Area R at 998.0E-3 M" * T h e le tte r " rn " in fh ie c o lu m n in d ic e te e th e ! rn e n u e l in te g ra tio n w e e p e r f o r m e d o n t h e t c o m p o u n d . Pnnt Date and Time: 10/18/2000 12:41 Page i 000683 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-67 Page 66 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621007.D Vial: 7 17 Oct 2000 15:48 Operator: MAN lx Std-6 Water Inst : MS7 MS Integration Params: rteint.p Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) 'Aburtanr ! 250* 2000, iI 1500j | 1000 TIC. 0621007.D A /i I 500) ! -, Tm*-> 6.10 6.20 6.30 6.40 MVL ____ -____ : Ion; 131 (7.143) | Resp: 4054 i ~ | 1 1 I ~ ' "l ........... 1 " ^ ' ^ 11 i 6SO 6.60 6.70 6 80 6 90 7.00 7 10 7.20 7.30 7.40 7.50 ___________________ Amount :_______0 . 0 0 ; r r r y r - r - , 7.80 7.70 7.90 7.90 _________________ 6.20 6.30 6.40 6.S0 660 6 70 6.80 6 30 7.00 710 7 30 7.30 7 40 7-50 7 60 7 70 7 80 7 90 -d4 Amount: 0.00 Ion: 135 (7.170) Resp: 4062 f\ i\ i :-- ----p - f -- !|-- !-- n- 1*r~T"pi-- |~'-I|ir i" "T''* - - T ' -y ; 6.20 6.30. 6,40..6.50.. 6.60 670 680 6.90 7 00 7 10 7.20 7.30 7 40 750 7.60 7.70 7.80 7.90 Q62107.D PABX BVA.M Wed OCt 18 12:41:59 2000 Page 2 0C0690 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-68 Page 67 Peak Area Report Sample Name: 1* Std-7 Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 16 08 Operator: MAN Data FileName: 0621008.D Instrument Name: MS7 Data File Path: D:'MSD_DATA\PAB\PA8X`BVA\1-62-1\ Instrument Vial Number: 8 Instrument Method Name: PA9X_BVA ConiDound # 1) 7) Compound Name MVL MVL-d4 RatonHon Tim (Min i 7.14 7 17 Ion 131 135 p M k Area M* 6314 3766 Peak Area Ratio 167 7E-2 M* * The tetter "m" in tei* column in d icai that manual Integration wee performed on that compound. Pnnt Date and Time 10/18/2000 12:42 Pag**: I 000691 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-69 Page 68 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621008.D Vial: 8 17 Oct 2000 16:08 Operator: MAN lx Std-7 Water Inst : MS7 MS Integration Params: rteint.p Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC: 0621006.0 I 2000^ i I 100! i\ ii j ; Il I! \ \ !v Tim-. 610 6.20 6.30 6 40 6.50 6.60 5.70 6.80 6.90 7.0 7.10 7.20 7 30 7.40 7 SO 760 7.70 7.00 7.90 MVL __ ________________________ Amount :_______0.00______ __ !" Ion: 131 (7".141) j Reap: 6314 / / \ V. 6.20 6.TO 640 6.50 _-d4_ Ioni 135 (7.169) Reap: 3766 6.70 6 80 6 90.. 7.00 7 10 7.20 7.30 " 7.40 7.50 7.60 7.70 7.B0 7.90 ' Amount: 0.00 !\ !\ \ \ r 6._20.6,30. 6,40..6.50 6.60 6 70 660 6.90 7 00 7.10 7.20 7. 7 40 7.50 7 60 7.70 7.B0 7.90 0621008,D PABX BVA.M Wed Oct 18 12:42:18 2000 Page 2 080692 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-70 Page 69 Peak Area Report Sample Name: 1x Control Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 16:27 Operator: M A N Data File Name: 0621009.D Instrument Name: MS7 Data File Path: 0 :WSD_DATA\PAB\PABX-BVA\1-062-t\ Instrument Vial Number: 9 Instrument Method Name: PABX_BVA Compound # U 2) Compound Name . MVL MVL-<J4 Retention Time (Mln.l 0.00 7.17 Ion 131 135 Pk Area M* 0 4595 Peak Area Ratio OOO.OE-3 M* The letter *m" in tnia column Indicate mat manual integration wea performed on mat compound. PrintOale end T>me 10/18/2000 12 42 Page 1 000693 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-71 Page 70 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621009.D Vial: 9 17 Oct 2000 16:27 Operator: MAN lx Control Hater Inst : MS7 MS Integration Params: rteint.p Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) jfliibundanee 2000! TIC:0621009.0 1500; /\ 1000: j\ j 500- i / I .... ................... ................... ! P ' r 'r ' ! 1 ~ 1 . | i , ' " . 1 i' i i . , , | . . - r ' r -- - i -- r-- -- 1 ........ Tm ~> 6 to 6.20 6.30 6.40 6.50 660 670 6.60 690 7.00 7 10 720 730 740 750 760 770 7BO 7.90 MVL ________________Amount: Not Found______________________ !' ion: 131 (0.000)..... ' I A-- . A >./^-' \ -J 6.20 6.30 6 40 650 6.60 6.70 L - d 4 ___________________ Ion: 135 (7.170) 1 Resp: 4595 690 7.00 710 720 7 30 740 750 760 7.70 7.80 7.90 Amount: ~ 0.00 I1 'r y~r~r~`~r'-i'-' 6 20 630 6.40 650 6.60 670 6.80 6.90 700 710 7 20 7.30 7.40 750 7 60 7,70 780 790 0621009.D PABX_BVA.M Wed Oct 18 12:42:37 2000 Page 2 0C0694 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-72 Page 71 Peak Area Report Sample Name: 1* O C HIGH Waiw Notebook Reference: PABX-BVA-1-062 Date Acqulied: 10/17/2000 16:47 Operator: MAN Data Rie Name: 0621010.D Instrument Name: MS7 Data File Path: D:\MSD_DATA\PA8iPABX-BVAVI-062-1\ Instrument Vial Number: 10 Instrument Method Name: PA8X_BVA Comoound 9 U 2) ComDound Name MVL MVL-dA Retention Time /Min i 7.14 7.17 ton 131 135 Peak Area M* 5366 4021 Peak Area Ratio 133.4E-2 M" " The terter "in" in thift column in d icai trial manual Intaglino#! waa pertormed on that compound. Print Data and Time: 10/18/2000 12:42 Pugn i OGGCS Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-73 Page 72 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621010.D Vial: 10 17 Oct 2000 16:47 Operator: MAN lx QC HIGH Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC-.0621010.0 A \ looo 6.10 6.20 JL3Q_._6.4p 6 50 6.60 6.70 6.80 6.90 7.00 7 10 Tip 7.30 740 I SO 760 770 780 7.90 MVL .... " "" Amount: 0.00 0621010.D PABX BVA.M Wed Oct 18 12:42:56 2000 Page 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-74 Page 73 Peak Area Report Sample Name: lx Q C MIO Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 17:07 Operator: MAN Data File Name: 0621011.D Instrument Name: MS7 Data File Path: D:\MSD_DATAVPABVPABX-BVA\1-062-H Inatrument Vial Number: 11 Instrument Method Name: PABX_BVA Comoound f 1) 2) Comoound Name MVL MVL-d4 Retention Time (Min i 7.14 7.17 Ion 131 135 Peak Atea M* 2034 m 4323 Peak Atea Ratio 470.5E-3 M" The letter "m" in this column indicates that manual integration was performed on that compound. Print Date and Time 10/18/2000 12:43 Page 1 06B7 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-75 Page 74 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621011.D Vial: 11 17 Oct 2000 17:07 Operator: MAN lx QC MID Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance 20001 TIC: 0621011.D AA 150o! ; 1000 50Q- r'r1 '1 1 Time- 6.1C 6.20 0.30 0 40 MVL Ion: 131 (7.142) Resp: 2034 M II I\ i; /v 'T r_ra*'"T . . . . . T--p , . P -.-. .TT-, 6.50 6.90 6.70 6.8Q 6.9Q 7.00 7.10 7.20 Amount: A 1 ............. . 7. 7.40 7.50 7.60 0.00 ' ` .......... - ! 7 70 7.80 7.90 /I 1l !\ V_ / 6.20 6.30 6 40 650 6.60 6.70 6.60 6.90 r.00 7.10 7 20 7.30 7.40 7.50 7 60..7 70 7.80 7.90 -d4 Amount : 0.00 Ion: 135 (7.169) ! Reap: 4323 j l "T - - *-- r -- - M ,r l ---r , , ,->-T r , , r-r-e-i-------------------------r-i-. . ........ ; . ..... . r . . , , r . - . : 6.20 6.30 6 40__6,50 6.60 6.70 6.80 6.90 7.00 7.10 7 20 7.30 7 40 7.50 7.60 7.70 7.80 7.90 | 0621011.D PABX_BVA.M Wed Oct 18 12:43:16 2000 Page 2 0QOC98 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-76 Page 75 Peak Area Report Sample Name: 1x O C L O W Water Notebook Reference: PABX-8VA-1-062 Date Acquired: 10/17/2000 17 2t> Operator: MAN Data File Name: 0621012.0 Instrument Name: MS7 Data File Path: D:\MSD_DATA\PABSPABX-BVA\1.062-1\ Instrument Vial Number: 12 Instrument Method Name: PABX_BVA Compound # 1) 2) Compound Name MVL MVL-04 Retention Time (Min.l 7 14 7.17 j0Q 131 135 Peak Area M* 680 m 4096 PaK A f a Ratte 166.OE-3 M* * The letter "m" m th u column mdteat that manual intagraUon w performed on that compound. Print Dele and Time: 10/18/2000 12:43 Page 818819 Primedica-Worcester Project number: PABX-BVA 418-018 -.PAGE 1-77 Page 76 Quantitation Report (QT Reviewed! Data Pile Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-Q62-1\0621Q12.D Vial: 12 17 Oct 2000 17:26 Operator: MAN lx QC LOW Water Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M IRTE Integrator) 0621012.D PABX BVA.M Wed Oct 18 12:43:36 2000 Page 2 000700 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-78 Page 77 Peak Area Report Sample Name: 1x O C LLOQ Water Notebook Reference: PABX-BVA-1-062 Dale Acquired: 10/17/2000 17.46 Operator: MAN Data File Name: 0621013.D Instrument Name: MS7 Data File Path: D:\MSD_OATA\PABVPABX-BVAU-062-1\ Instrument Vial Number: 13 Instrument Method Name: PABX_8VA Comoound # u 2) Comoound Name MVL MVL-4 Retention Time iMin.t 7 14 7 17 ton 131 135 Peak Area M ` 354 4755 Peak Area Ratto 744 5E-4 M* * Th tettar "hi" m this column indicates that manual integrali* was performad on that compound. Print Date and Time: 10/18/2000 12:43 Page 1 000701 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-79 Page 78 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621013.D Vial 13 17 Oct 2000 17:46 Operator MAN lx QC LLiOQ Water Inst MS7 MS Integration Parants: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance 20001 i i5oo; TIC 0621013 D 10001 i 500] /\ Time-> 610 6.20 6.30 640 650 6 60 670 6 BO 630 7 00 7.10 7 20 7.30 740 7SO__ 7J MVL ____ ___________Amount :___________________ 0.00 '" Ion: 131 (7 .~138l .......... !Resp: 354 7 70 780 790 6.20 5.30 640 6.50 .6.60 6.70 6.B0 6.90 7.00 7.10 7.20 7.30 7.40 7.50 760 7.70 780 790 d4 __________________ Amount : ~ 0. OCT Ion: 135 (7.170) Resp: 4755 !\ f\ !\ '''r'i'1` I 6.20 6.30.. 6.40 6.50 6.60 6.70 6.60 6.90 7.00 7.10 7.20 7.30 740 7.50 7.60 770 760 7.90 0621013.D PABX BVA.M Wed Oct 18 12:43:56 2000 Page 2 000702 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-80 Page 79 Peak Area Report Sample Name: 1x C HIGH Plasma Notebook Reference: PABX-8VA-1-062 Date Acquired: 10/17)2000 18 05 Operator: MAN Data File Name: 0621014 D Instrument Name: MS7 Data FilePath: O:\MSD_DATA\PAB\PABX-8VAU-062-1\ Instrument Vial Number. 14 Instrument Method Name: PABX_BVA Compound # 1) 2) Compound Name MVL MVL-04 Retention T w ? IMInJ 7.14 7 16 2 131 135 P u k Area M* 8537 6540 PeaK Area Ratio 130 5E-2 M* - The letter "m" in mi column indicate* that manual integration w performed on that compound. Print Dale and Time: 10/16/2000 12:44 Paq p. .1 000903 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-81 Page 80 Quantitation Report (QT Reviewed) Data Pile .Acq On Sample I Mise D:\MSD_DATA\PAB\FABX-BVA\l-062-l\0621014.D Vial: 14 17 Oct 2000 18:05 Operator: MAN lx QC HIGH Plasma Inst : MS7 MS Integration Parama: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundance 4000 3000 1 2000 TIC: 0621014.0 [\ i\ ; *, ii !i 1000 ! \ ; '' *T'"- i 1 '- 'r *T'i - ' , '1 1 1 i '7 i ' i ' ' j i Tirm-* 610 6.20 6 X 640 6.50 6.60 6.70 6.80 6 90 700 7.10 720 MVL " ~ Amount : r Ion: 131 (7.137) - :iResp: 8537 A :M . i ' '-' 7.30.7,40 7.50 7.50 6.0 : ',f 1 i ' 7 70 7 80 r "' ; 7,90 | ... I ' i \ i 620 6.30 6.40 6.50 6.60 6 70 6 80 6.90 7.00 7.10 7.20 7.X 7.40 7.X 7.60 7.70 7.80 790 ^^IVL-d4 Amount : 0 00 W r Ion: 135 (7 .161) ; Reap: 6540 A l\ I !\ ;\ 1 / \. 1 1' 1' 6.20 6.x 6.40 6.50 6.60 6.70 5.80 6.90 7 00 7 10 7.20 7.30 7.40 7.50 7.60 -j O > 7B0 7.90 ! !i f 0621014.D PABX BVA.M Wed Oct 18 12:44:15 2000 Page 2 000704 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-82 Page 81 Peak Area Report Sample Name: 1x Q C MID Plasma Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 10.25 Operator MAN Data FiteName: 0621015.D instrument Name: MS7 Data FitePath: D:\MSO_DATA\PAB\PABX-BVA\1-062-1\ Instrument Vial Number. 15 Instrument Method Name: PA8XJ3VA Comoound # 1) 2) Compound Name MVL MVl-d4 Retention Time (Min.l 7 14 7 16 Ion 131 135 Peak Area M* 2024 3888 Peak Area Ralio 507.5E-3 M* * The toner "m" m this column indicate* that manual integration was pertormed on that compound. Prml Date and Time. 10/18/2000 12:44 Page 1 007DS Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-83 Page 82 Quantitation Report (QT Reviewed! Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621015.D Vial: 15 17 Oct 2000 18:25 Operator: MAN lx QC MID Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) fAMmdanc 200& TIC: 0621015.0 j\ | 1500 ' 1000 !I i! i 500 /\ V T . -. T T ------. 6.10 6.20 6.30 6.40 MVL ~ ; Ion: "131 '(7.137) : Resp: 2024 6.SQ 6.60 'T < 6.70 6.80 V . T i'. i T I - r T T T , I ; . i .......................... - ......................... ' 6.90 7.00 7.10_ 7.20 7.30 7.40 7.SO 7.60 7.70 Amount: " 0.00 I ' 1 I 7.80 7.90 .\ I\ \ 620 6 .X 6.40 6.50 6.60 6.70__ 6.80 6.90 7,00 7.10 7.20 7 .X 7 40 7.50 7,60 7.70 7.80 7 . L-d4 ____ " Amount: 0.00 Ion: 135 (7.165) Resp: 3988 \ !I /\ /\ 6.20 6 30 6.40 6.50 6.60 670 6.60 6.90 7.00 7,10 7.20 . 7.30 7 40 7.50 7.60 __ 7.70__ 7.80 7.90 | 0621015.D PABX BVA.M Wed Oct 18 12:44:34 2000 Page 2 000706 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-84 Page 83 Peak Area Report Sample Name: 1x QC LOW Plasma Notebook Reference: PABX-6VA-1-062 Date Acquired; 10/17/200018 44 Operator: MAN Data Pile Name: 0621016 D Data File Path: D:\MSD_DATA\PA8\PABX-BVAV1-062-1\ instrument Method Name: PABX_BVA Instrument Name: MS7 Instrument Vial Number: 16 Compound # 11 2) Compound N an MVL MVL-d4 Retention Time fMin.l 7 14 7 16 &Q 131 135 Peak Area M" 1115 m 4700 Peak Area Ratio 237.2E-3 M* * T he letter "m" In th is colum n In d icates th at m anual integration w as perform ed on th at com pound. Pnnt Date and Time 10/18/2000 12:44 tags? i 000997 Primedica-Worcester Project number: PABX-BVA 418-018.-PAGEI-85 Page 84 Data File Acq On Sample Quantitation Report (QT Reviewed D: \MSD DATA\PAB\PABX-BVA\1-062-l\0621016.D Vial: 16 17 Oct"2000 18:44 Operator : MAN lx QC LOW Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) undanc i 200 15001 1000 ! 500) TIC 0621016.0 ! j i Ln ~> 6.10 6.20 6.30 6.40 6 50 6.60 6 70 6.80 ~5 90 ^ . T ^ T lO 7.20 7 .3 0 ._ M 0 . . 7 . . 7 . 7 70 7 60 7 90 mvl" " Amount: 6.00 ! Ion: 131 (7.139) i Resp: 1115 ! A i ( ------- r - ^ -- i i r - , --1~1-- -- ~r 1 6,20 6.30 6.40 6.50 6.60 d4 Ion: 135 (7.162).. :Resp: 4700 .r . -. -j ..............| ............... r . t' . .t- . - . -- .......... | -- ,-- i-- 1 i 1 -- ' 6.70 6.60 6 30 7.00 7.10 7.20 7.30 7 40 7.50__ 7,60__ 7 7 0 .... 7 60 7.90 _________________ Amount:____ _ _0 00____ iA . 6 20 6.30 6.40 6.50 6.60 6.70 6.60 6.90 7.0 7.10 7.20 7.30 7.40 7.50 7 60 7 70 7 80 7 90 0621016.D PABX BVA.M Wed Oct 18 12:44:54 2000 Page 2 000708 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-86 Page 85 Peak Area Report Sample Name: 1x Control Plasma Notebook Reference: PABX-BVA-1 -062 Date Acquired: 10/17/2000 19:04 Operator: MAN Data File Name: 0621017.D Data File Patti: D:\MSD_0ATA\PAB\PABX-BVA\1-062-n Instrument Method Name: PABX-BVA Instrument Name: MS7 Instrument Vial Number: 17 Comoound # 1) 2) Comoound Name MVl MVL*d4 Retention Time IMin.i 7.14 7 17 Ion 131 135 Peak Area M" 577 m 4860 Peak Area Ratio 1 M .7 E -3 M* * T h e letter "m" in th is colum n indicata* th at m anual integration w a s perform ed o n that com pound. Pnnt Date and Time: 10/18/2000 12:45 PiCJH 1 000709 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-87 Page 86 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621017.D Vial: 17 17 Oct 2000 19:04 Operator: MAN lx Control Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) lAbunoanc* TIC: 0621017. ! 2DOO; ii I: ' 1500' ii I| | 1000I 500; /i I\ ! 7*--- ;'! -- '-- r 1 ,-- 6.10 6.20 6.30 6-40 6.50 5 60 6.70 6.80 6.90 7.00 7 10 7 20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 o o o MVL Amount : I Ion: 131 (7.138) | Hasp: 577 j i\ ! !\ ! i\ _ ; 6.20 6.30 6.40 6.50 6 60 6.70 6.80 6.90 7.00 7.10 7.20 7.30 7 40 7.50 7.60 7.70 7.80 7.90 O o o |IVL-d4 Amount : f Ion: 135 (7.165) ! Reap: 4860 f) !\ |A 1 i t i 1 i i ; `1 j i j I 6,20.... 6.30 6 *0 6 50 6.60 6,70 8.80 6.90 7.00.... 7.10 7.20 . 7,30 7 . .....7.50 7.60 7.70 7.80 7.90 0621017.D PABX_BVA.M Wed Oct 18 12:45:13 2000 Page 2 000710 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-88 Page 87 Peak Area Report Sample Name: 2 x QC HIGH Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 19:24 Operator: MAN Data Fife Name: 0621018 D Data Fite Patti: D:\MSD_DATA\PAB\PABX-ftVAM-062-1\ Instrument Method Name: PABX_BVA Instrument Name; MS7 Instrument Vial Number: 18 CompouQ<)| n 1) 2) Compound Name MVL MVL-<W Retention TimolMin.) 7U 7 17 Ion 131 135 Peak Area M* 7314 5545 Peek Area Ratio 131.9E-2 M* * The latter "m" in t h ii colum n indicala* m ai m anual integration m m parform ad on m at com pound. Print Dale and Time 10/18/2000 12:45 Page- 1 000711 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-89 Page 88 Quantitation Report !QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621018.D Vial: 18 17 Oct 2000 19:24 Operator: MAN 2 x QC HIGH Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) lUbundane*' I 4000| TIC: 0621016.D 3000* 2000 1000: ;I ..... .----1 I : I m -- ......-- ' I'1' ' ' ---- ------------r ^ r ^ - 6.10 6.20 6.30 6.40 6.50 6 60 67 0 6.80 6.90 7.00 7.10 720 7.30 7.40 MVL ___________ ___ " ' ' Amount :______ 0.00 !.Ion: 131 (7.140) ............ . 'Reap: 7314 A M ;\ I\ 7.60 7.70 7.80 7.90 .S.20_ 6.30 6 4 0 6.50 6.60 6.70 6 80 6.90 7 . 0 0 ^ 7 10 7.20 ' 7.30 7.40 7,50 L-d4 __________ Amount : 0.00 Ioo:n: 135 (7.167) Reap: 5545 7.70 7 8 0 7.90 I J 6.20 6 30 6.40 6.50__ 6.60~ 6 7 0 j! i\ I\ i\ 'I" 1I=' 6 6 0 6 90 7 00 7.10 7 2 0 7 3 0 7 4 0 7.50 7 60 7 7 0 7 8 0 _ 7 .io ! 0621018.D PABX BVA.M Wed Oct 18 12:45:33 2000 Page 2 000712 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-90 Page 89 Peak Area Report Sample Name: 2 x QC MIO Walei Notebook Reference: PA8X-BVA-1-062 Date Acquired: 10/17/2000 19 43 Operator: MAN Data File Name: 0621019.D Data File Path: D \MSD_DATAIPAB\PABX-BVA\1-062-1\ Instrument Method Name: PABX_SVA Instrument Name: MS7 Instrument Vial Number: 19 Comoound # 1) 2) Compound Name MVL MVL-d4 Retention Time fMm.) 7.14 7 15 km 131 135 Peak Area M* 2025 m 4390 Peak Area Ratio 461.3E-3 M* = The teller "rrT in th is co lu m n indicate* th a t m anual integration w e t perform ed on th at com pound. Print Oate and Time 10/18/2000 12:45 Page l 000713 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-91 Page 90 Quantitation Repor (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621019.D Vial: 19 17 Oct 2000 19:43 Operator: MAN 2 x QC MID Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) toundance 2000 1500 1000 TIC: 0621019.0 i\ !I I! /; ;i ; : ' 500 ! \ 1\ \ Tim--> 6.10 6.20 6.30 6.40 6.iso 6.60 6.7D 6.B0 6.90 7.00 7.10 7.20 7.30 7 40 7.50 7 60 7 70 7.60 7.90 i MVL Amount: 0.00 Ion: 131' (7.137)' " " ... .1 Resp: 2025 ji I 6,20__ 6.30 6.40 5.SO 6,60 6.70 6.60 6.90 .....7.00 7.10 7.20 7 .30 7 40 7.50 7.60 7 70 7.B0 7.90 U V h -d4 _____________ Ion:'" 35' (7.164) _________ Amount : .00 Resp: 4390 _6.20__ .6 30 6.40 SSO 6 60 6.70 6 80 6.90 7.00 7.10 7.20 7 3 0 7 40 7 53 7 60 7.70 7 60 7 3 0 0621019.D PABX_BVA.M Wed Oct 18 12:45:52 2000 Page 2 000714 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-92 Page 91 Peak Area Report Sample Name: 2 x OC LOW Water Mnttehnalr Bufemneii' PARJ1UA.1UM? Date Acquired: 10/17/2000 20:03 Operator: MAN Data File Name: 0621020.D Data File Path: D:\MSO_DATA\PAB\PA0X-0VA\1-O62.1\ Instrument Method Name: PABX_BVA Instrument Name: M57 Instrument Vial Number: 20 ComDOund * 1) 2) Coimxnind Name MVL M V l-4 4 Retention Time (Min,) 7 14 7 17 Ion 131 135 Peak Area M` 767 m 4380 Peak Area Ratio 175.1E-3 M* The tetter "m" in H ut co lu m n In d icates th at m anual integration w as p erfo rm ed o n th at com pound. Print Data and Time: 10/18/2000 12:46 Page. .1 000715 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-93 Page 92 Quantitation Report QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621020.D Vial: 20 17 Oct 2000 20:03 Operator: MAN 2 x QC LOW water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) 1 2000! ! 1500; 1000; 1 500; f --i - r j '-- ---r----T--1--' 1 T-n'i--h-- iTinw- 61P 6.20 6 30 6.40 6.50 6.60 6.711 6.80 MVL_ ____ ______ _____ Ion: 131 (7.139) : Resp: 767 TIC: 0621020.0 A I; !I I i ( j , ............1 1 1 1 ........................--T -'"I--'f'....-- 6.90 7.00 7.10 7 3 ) 7 30 7 . 7.50 7.60 7.70 7 80 7.90 Amount: 0.00 !\ I\ " n -- 1T^ i " 1 ' I-- 1-- T ' -'' ' : 1-- 111 111. ....... I-- --- 'm^ 1* 620 6 30 6 40 6.50 6.60 6.70 6.B0 6.90 7 .X 7 10 7.20 7 30 7 40 7.50 7.60 7 70 7 60 7.90 -d4 Amount: 0.00 Ion": 135 (7.167) Resp: 4380 J\ /v i_6 20. . 6 30 6 4 0 6 50......6.60 6.70 6.80 ' ' 6 90 7 00__ 7 J 0 ___ 7.20 7.30 __ 7 40___ 7.50 7 60 770 7 80 7.90 0621020.D PABX BVA.M Wed Oct 18 12:46:12 2000 Page 2 000716 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-94 Page 93 Peak Area Report Sample Name: 2 x QC II O Q Water Notebook Reference: PABX-BVA-1-Q62 Date Acquired: 10/17/2000 20.22 Operato: MAN Data File Name: 0621021.D Data Fite Patti: D:\MSD_DATA\PA8\PABX-BVAM-062-1\ Instrument Method Name: PABX_BVA Instrument Name: MS7 Instrument Vial Number: 21 ComDound# 1) 2) Comoound Name MVL MVLkM Retention Time (Min.l 7 14 7.17 Ion 131 135 Peak Area M* 361 m 5175 Peak Area Ratk 697.6E-4 M' * The letter ,'m" in Ihi* colum n indicate* m at m an u al in teg ratio n w a i perfo rm ed o n that com pound. Print Date and Time 10/18/2000 12:46 Pag 00071*7 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-95 Page 94 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621021.D Vial: 21 17 Oct 2000 20:22 Operator: MAN 2 x QC LLOQ Water Inst : MS7 MS Integration Params: RTEINT.F Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) 0621021.D PABX_BVA.M Wed Oct 18 12:46:31 2000 Page 2 00071 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-96 Page 95 Peak Area Report Sample Marne: 2 x QC HIGH Plasma Notebook Reference: PAJBX-BVA-1-062 Date Acquired: 10/17/2000 20:42 Operator: MAN Data Frie Name: 0621022.D Data Frie Patti: O \MSD_OATA\PAB\PAaX-VAU.0G2-1I Instrument Method Name: PABX_BVA Instrument Name: MS7 Instrument Vial Number: 22 ComDound # 1) 2) ComDound Nam MVL MVL-d<l Retention Tima (MinJ 7.13 7.16 k>n 131 135 P u k Arca M* 6581 5208 Peak Area Ratio 126.4E-2 M* The latter "m" ln m is c o lu m n in d c a la s ttvat m anual Integracin w *s parform ed o n Chat eom pound. PnntDate andTime: 10/16/2000 12:46 l'ge* \ 000719 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-97 Page 96 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621022.D Vial: 22 17 Oct 2000 20:42 Operator: MAN 2 x QC HIGH Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundance 3000; TIC: 0621022.D IA \ ! i2000: j \1000j . --- 6.10 5 2 0 6.30 5.40 6.50 6.60 6 70 6.80 6.90 7 0 0 7 10 7.20 7.M 7 40 7.^0 7.60 7.70 7.B0 7,90 MVL Amount: 0.00 Ion: 131 (7.134) ^ Resp: 6581 : i \ ,, 1 66-.2210 6-30 6.40 6.50 6.60___6.70 6.60 6.90 7.00 7.10 7 20 7.30 7 40 7.50 7.60 7 70 7 80 7 90 -d4 Amount: 0.00 Ion: 135 (7.162) \ Reeisp: 5208 !\ I --------- ---- -------- !-----:--- -------- ------r---- - - ' ....... ---------------------- -- - y-,-r-P ;'' -i--- - ----- -- 6.20 6.30 6 40 6.50 S.60 6.70 6 80 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 ----- 1" ' 1 ! 1 7.70 7.80 7.90 0621022.D PABX_BVA.M Wed Oct 18 12:46:51 2000 Page 2 900720 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-98 Page 97 Peak Area Report Sample Name: 2 x QC MID Rasma Notebook Reference: PABX-8VA-1-062 Date Acquired: 10/17/2000 21:01 O perator: MAN Dale File Name: 0621023 D Data File Patti: D:\MSD_DATA\PAB\PABX-BVA\i -062-1\ Instrument Method Name: PABX-BVA Instrument Name: MS7 Instrument Vial Number: 23 Comoound # i) 2) Compound Name MVl MVL-04 Retention Time (Min.l 7.13 7.16 &> 131 135 Peak Area M! 2280 m 4580 PlfcAfi Ratio 497.8E-3 M' * Th letter "m" in tht* c o lu m n in d icates th at m anuel integralton w as perform ed on th a t com pound. Pont Date and Time: 10/18/2000 12.47 Page L 000721 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-99 Page 98 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621023.D Vial: 23 17 Oct 2000 21:01 Operator: MAN 2 x QC MID Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundance 2000- TIC: 0621023.D /; ,: j 1500; j ' 1000; : soo| I\ I; /\ I -Tr.-r-rr-p------m Tm -> 6 10 6.20 6.30 .6.40 MVL 6.50 >- , - ; , , t . . t " . . . 6.60 6.70 6.B0 6.90 7.00 7.10 7:20 _ Amount : i i ... . - ' i ..... r- ................. 7 30 7-40 7.50 7 60 7 70 7 80 7.90 0.00 _________ ! Icm7~131 (7.134) iResp: 2280 A i 6.20 6.30 6.40 6.50 6.60 to fL - 'i ' IIcon: 135 (7.162) j Resp: 4580 6.70 6.60 6 90 . 7 00 770___7.20 7.30 _ _ _ ......y . Amount: 7 40 7.50 C .00 7.60 7.70 7,80 7.90 " _____________ j_6,20 6.30 6.40 6.50 6 60 6.70 6.60 6.90 7.00 710 7.20 7.30 7 40 7.60 7.60 7.70 7.60 7.90 0 6 2 1 0 2 3 .D PABX_BVA.M Wed O c t 18 1 2 : 4 7 : 1 0 2 0 0 0 Page 2 000722 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-100 Page 99 Peak Area Report Sample Name: 2 x QC LOW Ptaema Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 21:21 Operator: MAN Data File Name: 0621024D Data File Path: D:\MSD_DATAVPAatPAtX-B VAU-062-11 Instrument Method Name: PABX_BVA Inatnimant Name: MS7 Instrument Vial Number: 24 Compound * 1) 2) Compound Name MVL MVL-d4 Retention Time fMin.l 7.13 7.18 k>n 131 135 Peak Area M* 996 4321 Paak Area Ratio 230.5E-3 M" " The letter "m" in th is colum n in d ic a te s th at m an u al in teg ratio n w a s p a tlo n n a d on th at compound. Print Date and Time: 10/18/2000 12:47 Page i 000723 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-101 Page 100 Quantitation Report (QT Reviewed! Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621024.D Vial: 24 17 Oct 2000 21:21 Operator: MAN 2 x QC LOW Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance i 2000^ 1S00: 1000 TIC: 0621024.D | f I i ! 500 jrinw~> 6.10 MVL 6.20 --------------1-----1,.,-r-r^-T---r^-7-r^- 6 .X 6 40 6.50 6.60 6.70 ~ ; Ion: 131 (7.134) i Reap: 996 6.B0 / , T n 7 f |- ' .-vr , . . . 6.90 7 00 7.10 7.20 Amount: t-------t*---------,-----,------------------- ! 7.30 7.40 7.50 7,60 7.70 0.00 ' I 1 7.80 7.90 I. i 6.20 6 30 6 40 6 50 6 60 ^fcrfVL_-d4 ' _ Ion : 135 !7.1621 !Resp: 4321 6 70 6 80 6 90 7 00 7.10 7.20 7 30 _ _______Amount': ..... ~ 7.40 7.50 7.60 7.70 0 00________ 7.80__ 7?0_ _ 6.20 6.30 6.40 6.50 6.60 6 70 6.80 6.90 7.00 7 10 7.20 7.30 7.40 7.50 7.S0 7.70 7.80 7 90 0 6 2 1 0 2 4 . D PABX BVA.M Wed O c t 18 1 2 : 4 7 : 3 0 2 0 0 0 Page 2 000724 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-102 Page 101 Peak Area Report Sample Marne; 2 x Control Plasma Notebook Reference: PABX-BV/A-1 -062 Dele Acquired: 10/17/2000 21:40 Operator: MAN Data File Name: 0621025.D Data File Path: D:\MS0_DATA\PABNPABX-BVA\1-062*1\ Instrument Method Name: PABX_BVA Instrument Name: MS7 Instrument Vial Number: 25 expound 1) 2) Compound Name MVL MVL-4 Retention Time IMin.1 7 13 7.16 Ion 131 135 Peak Area Ma 525 m 4463 Peak Area Ratio 117BE-3 M* The te tte r "w " in th is co lu m n in d icate th a t m an u al integration w as perform ed on that com pound. Print Date and Tima 10/16/2000 12 47 Page 1 000725 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-103 Page 102 Quantitation Report (QT Reviewed) Data File Acq On Sample Miao D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621025.D Vial 25 17 Oct 2000 21:40 Operator MAN 2 x Control Plasma Inst MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance TIC: 0621Q2S.O 2000* i 1500i h ii i1 i' | 1000' :! ! 500 I! i rI!TIiimmee----> 6O.1I0U 6g..2y0 1 T ' , rv ' `-T s-r-r-r-rT.i, -T, 11 ' 6o.3ju0 6,40 p6.g5y0 g6.6w0 w6.7<u0 g6..o6u0 6.90 7f .00 /| ! *\ !\ 1` ' 7r..1tw0 7..2c0v .T -TT.-.-- - - r . , . 7r .w3w0 7< .40 7.50 I 7.60 t '-rrr-rr7.70 7.60 7.90 MVL Amount: 0.00 - 0 6 2 1 0 2 5 . D PABX BVA.M Wed O c t 16 1 2 : 4 7 : 4 9 2 0 0 0 Page 2 000726 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-104 Page 103 Peak Area Report Sample Name: 3 * QC HIGH Watt Notebook Reference: PABX-8VA*1*0G2 Dele Acquired: 10/17/2000 22:00 Operator: MAN Data Fite Name: 0621026.0 Data File Path: D:\MSD_0ATA\PAB\PABX-BVA\1 -062-11 Instrument Method Name: PABX BVA instrument Name: MS7 Instrument Viet Number: 26 Compound # 1) 2) Comoound Name MVl M V l-0 4 Retention Tima (Min.) 7 14 7 16 Ion 131 135 Peak Area M* SS11 4246 Peak Area Ratio 129.8E-2 NT T he letter in th is colum n in d icate* m a t m anual integration w u p erfo rm ed o n th at com pound. Pnnt Date and Time: 10/16/2000 12:48 Page 1 000727 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-105 Page 104 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BVA\1-062 -1\0621026.D Vial: 26 17 Oct-2000 22:00 Operator: MAN 3 x QC HIGH Water Inst : MS7 MS Integration Parama: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundarte* 30001 TIC 0621026.D 2000* Tm-> 610 6-20 6.30 6 40 6.50 6.80 670 MVL ________ Ion: 131 (7.138) Reap: 5511 6.a0 8 90 7 CO 7 10 7.20 7.30 7.40 7.50 Amount: ' 0.00 7.60 77 0 7.80 7.90 : 6.20 6.30 8 40 6.50 6.60 L_-d4 ________ Ion: 135 (7.162) Reap: 4246 |-- ................. 6 70 S.B0 6 90 7.00 - T .......................... .... 1 7 10 720 7.30 Amount : i'' ....... ' ' " - I ' ' ' 7.40 7,50 7.60 7.70 7.80 0.00 n -- -- i l 7.90 \ ----- --- "T-- 11 ~rT~'V '`IT'- 'I"" -- f" I 6.20 6.30 6.40 6.50 6.60 6.70 600 6.90 7.00 7 10 720 7 3Q 7 . 7.50 7 60 7.70 7.80 _7_90 0 6 2 1 0 2 6 .D PABX_BVA.M Wed O c t 18 1 2 : 4 0 : 0 8 2 000 Page 2 00072 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-106 Page 105 Peak Area Report Sample Name: 3 x QC MID Water Notebook Reference: PABX-8VA-1-062 Date Acquired: 10/17,7000 22 19 Operator: MAN Data Fite Name: 0621077. D Data Fite Path: D:\MSDJ3ATA\PA8\PABX'BVA\1 ^062-1\ Instrument Method Name: PABX BVA Instrument Name: MS7 Instrument Vial Number: 27 Comoound # U 2) Compound Name MVL MVL-d4 Retention Tipw ttfipj 7.14 7 17 Ion 131 135 Peak Area M* 2059 m 4522 Peak Area Ratio 455.3E-3 M ' T he tetter "m" in tht* colum n in d icate* UuM mervual in teg ratio n w ae perfo rm ed o n th at com pound. Punt Date and Time: 10/18/2000 12:46 Page 1 000729 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-107 Page 106 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621027.D Vial: 27 17 Oct 2000 22:19 Operator: MAN 3 x QC MID Water Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) JxndOToe 2000! 1500* 10001 TIC: 0621027 D A !\ ! 1l i i `rTr` ' Tim- 6 10 6-20 MVL 1 ( 1 - r'-T't 6.30 6 *0 6.50 6.60 >f 670 6.80 ( ; "nlnnri 6 90 7.00 7.10 7.20 Amount: I , . |' i"i1''1 , rrr-rm-p-rt----'---------r-71- 7 30 7.40 7.50 7.60 7 70 7.80 7 90 0.00 0 6 2 1 0 2 7 .D PABX_BVA.M Wed O c t 18 1 2 : 4 8 : 2 8 2 0 0 0 Page 2 000730 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-108 Page 107 Peak Area Report Sample Name: 3 x QC LOW Water Notebook Reference: PABX'GVA* 1-062 Date Acquired: 10/17/2000 22:39 Operator: MAN Data File Name: 0621028 D Data File Path: DAMSD_DATA\PAB\PABX-BVA\1-0e2.1\ Instrument Method Name: PABX_BVA Instrument Name: MS7 Instniment Vial Number: 28 ComDound # 1) 2) ComDound Name MVL MVL-d4 Retention Time IMin.l 7.14 7 16 Ion 131 135 Peak Area M" 930 m 5310 Peak Area Ratio 175 1E-3 M* * T he letter "m " m th is co lu m n in d ic a te s th at manusrf integration w as perform ed o n m a t com pound. Print Date and Time 10/18/2000 12 4B P*qe 1 088^31 Priinedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-109 Page 108 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621028.D Vial: 28 17 Oct 2000 22:39 Operator: MAN 3 x QC LOW Water Inst : MS7 MS Integration Params : RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) 0 6 2 1 0 2 8 .D PABX_BVA.M Wed O c t 18 1 2 : 4 8 : 4 8 2 0 0 0 Page 2 000732 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-110 Page 109 Peak Area Report Sample N am e 3 * QC LLOO Water Notebook Reference; PABX-BVA-1-062 Date Acquired: 10/17/2000 22.58 Operator: MAN Data File Name: 0021029.D Data File Path: D:\MSD_0ATA\PAB\PABX-SVAi1-0e2-1\ Instrument Method Name: PABX BVA Instrument Name: MS7 Instrument Vial Number: 29 ComDOund # 1) 2) Comoound Nama MVL MVL-d4 Retention Tima (Min.] 7 14 7 16 OQ 131 135 P * A r w M* 353 47S9 Peak Area Rabo 737. IE-4 M* " The tetter "m" in thie colum n in d c e le s ihel m anual integration eras parform ad o n th at com pound. Print Dale an d Time: 10/16/2000 12:48 Page l 000733 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-111 Page 110 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BVA\1-062-1\0621029.D Vial: 29 17 0ct~2000 22:58 Operator: MAN 3 x QC LLOQ Water Inet : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundance 2000i TIC: 0621029.0 A j\ | 1500,1 ;i I 1000*1 ;t : 5oo; / 6.10 620 6.30 6.40 6 50 6.60 6.70 680 6.90 7.00 7 10 7,20 7.30 7.40 7.50 7.60 7.70 7.60 7.90 MVL Amount: 0.00 Ion: 131 (7.137) Resp: 353 A 1! i: i i1 i ___ /i__\_ 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7 00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.60 7.90 ^ V L -d 4 Amount: 0.00 Ion: 135 (7.164) i Resp: 4789 A i T ~r 6.20 6.30 6 40 6 SO 6 60 6.70 6.80 6.90 7.00 7.10 7.20 7 30 7.40 7,50 7.60 7.70 7.80 7.90 0 6 2 1 0 2 9 . D PABX BVA.M Wed O c t 18 1 2 : 4 9 : 0 7 2 0 0 0 Page 2 000734 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-112 Page 111 Peak Area Report Sample Nam: 3 1 QC HIGH Plasma Notebook Reference: PASX-BVA-1-062 Date Acquired: 10/17/2000 23 18 Operator: MAN Data File Name: 0621030.0 Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1 -062-1 \ Instrument Method Name: PABXJBVA Instrument ama: MS7 Instrument Vial Number: 30 Compound * 1) 2) Compound Name MVL MVL-4 Retention ItfflUM ifel 7.13 7.16 tSQ 131 135 Peak Area M* 6347 5002 Peak Araa Ratio 126 9E-2 M* T h e latter "m " In th is colum n in d ic les th at m anual in tag ration w a s perform ed on m at com pound. Pnnt Date and Time. 10/18/2000 12 49 1 000735 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-113 Page 112 Quantitation Report (QT Reviewed ) Data File : D:\MSD DATA\PAB\PABX-BVA\l-062 -1\0621030.D Vial: 30 Acq On : 17 Oct 2000 23:18 Operator: MAN . Sample : 3 x QC HIGH Plasma Inst : MS7 | Mise MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Adbunoanct TIC: 0621030 D f\ 300o!1 200Q] '\ j! 1000 !\ j^ ^ : - t . .-- ... J'r Tin- 610 6.20 6 30 6.40 6.50 6 60 6.70 6.80 6 90 7 00 7.10 7.20 7.30 740 750 7.60 7.70 7.80 7.90 MVL Amount : 0.00 Ion: 3 7.134) Resp: 6347 A i\ A I r" ! T - ' I --------n -- 6.20 6.30 6.40 6.50 6.60 6.70 N -d4 Ion: 135 (7.161) Resp: 5002 -------- r - -- 1-- I-- " -- I 1 1 I 1 1 : -- r~T~>-- -- ! ........ ' - i -- - t " 6.B0 6.90 .....7 00 7.10 7.20 7.30 7 <0 7.50 7.60 7,70 7.80 Amount : 0.00 r, i: A 7.90 1' I ' ' 6.30. 6.40 6 50 6.60 6.70 6.60 6.90 7.00 7.10 7.20 7 JO 7 40 7.50 7.B 7.70 7.80 7 90 0 6 2 1 0 3 0 .D PABX_BVA.M Wed O c t 18 1 2 : 4 9 : 2 6 2 0 0 0 Page 2 000726 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-114 Page 113 Peak Area Report Sample Name: 3 a QC MID Plasma Notebook R eference: PABX-BVA-1-062 Date Acquired: 10/17/2000 23 37 Operator: MAN Data Pile Name: 0621031.0 Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1-062.1\ Instrument Method Name: PABX_BVA Instrument Name: MS7 Instrument Vial Number: 31 Compound * 1) 2) Compound Name MVL MVLd4 Retention Tim (Min.) 7 14 7.16 Ion 131 135 Peak Area M* 2485 4925 Peak Area Ratio 504.6E-3 NT The letter "m " in th is colum n in d ic a te s th at m anual m teu rau o n w as p erto rm ed on that com pound. Print Date and Time: 10/18/2000 12:49 Cage 1 000737 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-115 Page 114 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVAU-062-l\0621031.D Vial: 31 17 Oct 2000 23:37 Operator: MAN 3 x QC MID Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundanc* i 20001 i TIC: 0621031.0 iAi /; i ; i \1000; i[ 6.10 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 720 7. 7.40 7.S0 7.60 7.70 7.80 7 90 MVL Amount: 0.00 Ion: 131 (7.137) Resp: 2485 !\ ' 6.20 6.30 6 40 6.50 L-d4 I f *.Ion: 135 (7.164) Resp: 4925 /\ !\ 6.70 6.80.. 6.90 700 7.10 7.20 7.30 7 40 7-50 7 60 7 70 7.80 790 Amount : 0.00 6.20 6 30 6.40 6.50 6.60 6 70 6 80 6.90 7.00 7 10 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 0 6 2 1 0 3 1 .D PABX_BVA.M Wed O c t 18 1 2 : 4 9 : 4 6 2 0 0 0 Page 2 000728 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-116 Page 115 Peak Area Report Sample Name: 3 *Q C L O W Plasma Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 23:57 Operator: MAN Data File Name: 0621032.D Inatniment Name: MS7 Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1-062-1\ Instrument VialNumber: 32 Instrument Method Name: PABX^BVA Compound # K 2} Compound Name MVL MVL-4 Retention Time IMin.1 713 7 16 ton 131 135 Peak Area M* S61 3B50 Peak A m a Rate 2236E'3 M* The letter 'W in thi column indicate that manual integration was performed an that compound. Print Data and Time: 10/16/2000 12 49 Page 00073 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-117 Page 116 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621032.D : 32 17 Oct 2000 23:57 Operator: MAN 3 x QC LOW Plasma Inst : MS7 MS Integration Parama: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) j*burvoance TIC: 0621032 D 'i iSOOj i 1000I a i\ 500; ---- - "1. '11-. i-, .r > '`(''''1''''1r ... : '-r-l-T'T-' ..i1i... *. <| .; fTim*-> 6.10 6.20 6.30 6 40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7.X 7 40 7.50 7.60 7.70 7.80 7.90 MVL A m o u n t: 0.00 Ion: 131 (7.134) 1Resp: 861 ; - L-------- - -- - ------ - . -- ---- --- ` 1-- r-|-r-v-- .-- ,-r-rrrr-r- f,rr 1 6.20 6 30 6 40 6.50 6.60 6 70 6.80 6.90 7. 710 7.20 7.30 7.40 7.50 7.60 7.70 7.B0 7.90 4tVL-d4 Amount 0. 00 Ion: 135 (7 .162) : Resp: 3850 A i . j /i \ \> ii 1 6.20 6 30 6.40 6Q 6.60 6.70 6.80 6.90 7.00 7.10 7 20 7.X 740 7.50 7.60 7 70 7.80 790 0 6 2 1 0 3 2 .D PABX_BVA.M Wed O c t 18 1 2 : 5 0 : 0 5 2 0 0 0 Page 2 00074 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-118 Page 117 Peak Area Report Sample Name: 3 Control Plasma Notebook Reference: PABX-BVA-1-062 Date Acquino: 10/18/2000 0 16 Operator: M A N Data FileName: 0621033.0 IrtstrumentMarne: MS7 Data File Patti: D VMSD_0ATA\PAB\PA8X-BVA\1-062-1\ Instrument Wial Number. 33 Instrument Mettvod Name: PABX BVA Compound # 1) 2) Compound Name MVL MVL-d4 Retention Time IMn.l 7.14 7 15 Ion 131 135 Peak Area M* 581 m 4846 Peak Area Ratio 119.9E-3 M- * The letter "m" in this column indicates that manual integration was performed on that compound Print Date and Time: 10/18/2000 12:50 Page 1 009741 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-119 Page 118 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621033.D Vial: 33 18 Oct 2000 00:16 Operator: MAN 3 X Control Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) TIC: 0621033.0 2000 1500 I t 1000 500 6 10 6.20 6-30 6 40 6.50 6.50 6.70 6 B0 MVL ! o t T'131 "{7.137 ) i Resp: 581 6.90 7~0T 710 7,20 7.30 7.40 7.50 '^ 6 0 "Amount: 0.00 7.70 7.B0 7.90 '----r ' 1 I 1 1 ' 'T'- I ---r-j-, -, -, , ------ - "TT''' 6.20 5.30 6.0 6.50 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 760 7.90 L-d4 Amount 0.00 m Ion: 135 (7.161) Resp: 4846 I' !\ 1-- I $20 6 30 640 6.50 660 .6.70 6.80 6.90 7.00 7 10 7 20 7,30 7.40 7.50 7.60 7.70 7.8 7.90 0 5 2 1 0 3 3 .D PABX_BVA.M Wed O c t 18 1 2 : 5 0 : 2 4 2 0 0 0 Page 2 000745 Primedica-Worcester Project number: PABX-BVA 418-018 .'PAGE 1-120 Page 119 Peak Area Report Sample Name: 4 x O C HIGH Water tiintokvuik Reference: PARX-flVA-1-062 Date Acquired: 10/18/2000 0 36 Operator: MAN Data FHe Name: 0621034.0 Instrument Name: MS7 Data Fite Path: D:\MSDJ>ATANPAB>PAeX-eVAV1-062-1t Instrument Vial Number: 34 Instrument Method Name: PABX-BVA Compound # it 2) Compound Name MVL MVL-4 Retention Tim IMtpJ 7.14 7 16 Ism 131 135 Peak Area M* 4620 3568 Pak Ana Ratio 129.SE-2 M* * The tetter "itT in this column indicale* that manual integration was performed on mat compound. PrintDate and Time 10718/2000 12:50 Page 1 000743 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-121 Page 120 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621034.E Vial: 34 18 Oct 2000 00:36 Operator: MAN 4 x QC HIGH Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) A btrKtenc* ! 2500 2000 TIC:0621034 0 1500, iooo 500 I 6.'lQ' 6.20~J 6.30 6.<*0 60 6.60" 6.70 6.60' 6.90'rot) ~ 7^0^730 7 30 7 40 7.50 7 60 7.70 7.80 7 90 MVL " _________________ Amount :________ 0 . 0 0 ________________________ Ion: 131 (7.139) Resp: 4620 iIA\t M JV i $.20 6.30 6 6,50. 6,60 ^feuVL-d4 W ion: 135(7.162) Resp: 3568 6.70 6.80 6.90 7.00 ............ ' 710 720 7.30 740 780 7 60 7.70 7.80 7 90 Amount :_______ 0.00___________________ rT"r~'"' I' ' , ! . . . . . . . . --------------p _ ---------n 6 20 6.30 6.40__ 6.SO 660 6 70 6 60 6 90 7 00 7.10 7.20 7 30 7 40 7 50 7-60 7 70 7 60 7 90 0 6 2 1 0 3 4 . D PABX BVA.M Wed O c t 18 1 2 : 5 0 : 4 4 2 0 0 0 Page 2 000744 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-122 Page 121 Peak Area Report Sampie fiant: 4 xQ C MID Water Notebook Reference: PABX-0VA-1-O62 Date Acquired: 10/18/2000 0:55 Operator: M A N Data FileNam: 0621035.D Instrument Name: MS7 Data File Path: D:\M5D_DATA\PABVPABX-BVAU-062-1\ Instrument Vial Number; 35 Instrument Method Name: PABX__BVA Comoound # 1) 2) Comoound Name mvl MVL-4 Retention Time fMin.l 7.13 7 16 Ion 131 135 Peak Area M* 2193 m 47T7 Peak Area Ratio 459.1E-3 M* * The Iettar ~m" in thU column indicate m at manual integration waa performed on that compound. Pnnl Date and Time: 10/16/2000 12:50 Pag* 1 00074 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-123 Page 122 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621035.D Vial: 35 18 OCt 2000 00:55 Operator: MAN 4 x QC MID Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) t a OTKjaOC* : 2S00] 2000| 1500; ; 1000; I 500: TIC: 06213S.D A I1 I\ /' j\ Tut --> 6 10 6.30 6.40 '.SO 6.80 670 68C MVL ! Ion: 131 (7.133) .. i Resp: 2193 ! 69C T 00 ?T 7.20 7.30 7~<K) 7.50 7.60 7.70 Amount': ... ~ 0.00 A 7.ag_j.9g___I IV 1'F." .f 6.20 6.30 6.40 6.50 6.60 6.70 6 60 6.90 7.00 710 7.20 7.30 7.40 7.50 7 60 7.70 780 7.90 2_d4 _ ___ Amount : 0.00 Ian:" 135 (7".161) Resp: 4777 !I n 6.20 6.30 6.40 6.50 6 60 6.70 6.80 6.90 7 00 7 10 7 20 7 30 7 40 7 50 7.60 7.70 7.80 7.90 0 6 2 1 0 3 5 . D PABX BVA.M Wed O c t 18 1 2 : 5 1 : 0 3 2 0 0 0 Page 2 00074S Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-124 Page 123 Peak Area Report Sample Name: 4 * Q C L O W Water Notebook Reference: PA8X-8VA-1-062 Date Acquired: 10r18/2000 1:15 Operator: MAN Data File Name: 0621036.0 Inatrument Name: MS7 Data Fite Path: D:\MSD_DATA\PA8\PABX-BVAtt-062-1\ Instrument Vial Number: 36 inatrument Method Name: PABX_BVA Comoound # H 2) Comoound Name MVL MVL-CJ4 Retention Time (Min.l 7 14 7.16 jSn 131 135 Peak Area M* 835 4966 Peak Area Ratio 1BB 1E-3 M* The letter "m" in thia column tndtcale* that manual Integration w et performed on that compound. Print Date and Time: 10/16/2000 12:51 Page 1 000747 Piimedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-125 Page 124 Quantitation Report (QT Reviewed Data File Acq On Sample Mise D:\MSD_ DATAN PAB\PABX-BVA\1- 062-1N0621036.D Vial: 36 18 Oct""2000 1:15 Operator: MAN 4 X QC now Water Inst MS7 MS Integration Parama : RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundance 2500; ! 2000| TIC: 0621036 D A i 1500; i 1000i 500. i.* ii \ 1\ 6.10 6.20 6.30 6 40 6.50 6.60 6.70 6.80 6 90 7.00 7 10 7.20 7.30 7.40 7.50 7.60 7 70 7.80 7.90 MVL Amount: 0.00 i Ion: 131 (7.136) j Reap: 835 i i ! 1A1 ! j\ ' 6.20 6.30 6.40 6.50 6.60 6.70 680 6.90 7.00 7 10 7.20 7.30 7.40 7.50 7.60 7.70 780 7.90 ktVL-d4 Amount: 0.00 r Ion: 135 (7.163) j Resp: 4968 I A 6.20 6.30 6.40..6.50 660 670 6.80 6.90 7.00 7 10 7 20 7 30 7 40 7 , 5 0 7.60 7.70 7 B0 7 90 0 6 2 1 0 3 6 .D PABXJ3VA.M Wed O c t i 8 1 2 :5 1 : 2 2 2000 Page 2 000748 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-126 Page 125 Peak Area Report Sample Name: 4 x Q C LLOQ Water Notebook Reference: PABX-BVA-1 -062 Date Acquired: 10/18/2000 1:34 Operator: MAN Data Fite Nemo: 0621037.D Instrument Name; MS7 Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1-062-n Inatmment Vial Number: 37 Instrument Method Name: PABX-BVA Compound 1) 2) Compound Name MVL MVl-d4 Retention Time (Min.) 7 14 7.16 Ion 131 135 Peak Area M* 365 5012 Peak Area Ratio 72B.3E-4 M* Tha lattar "m" In ttv t column indicalo that manual integration was partormad on that compound. PnntDate and Time: 10/18/2000 12:51 Page 1 000749 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-127 Page 126 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BVA\l-062-l\0621037.D Vial: 37 18 Oct~2000 1:34 Operator: MAN 4 x QC LLOQ Water Inst : MS7 MS Integration Params: RTBINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M {RTK Integrator) Atamdenca i 2500J 2000 ' TIC:062137.0 I I ; 10U0- j\ ! S00; j\ V1'*-'"- .... .-.:''f 1'.T-------- .......~ !---. T m - > 6.10 6.20 6.30 64C 6.S0 8.60 6.70 6-BO 6.90 703 7.10 720 7 30 7 40 7.50 7,60 770 7.B0 7SO MVL Amount: 0.00 0 6 2 1 0 3 7 .D PABX_BVA.M Wed O c t 18 1 2 : 5 1 : 4 2 2 0 0 0 Page 2 000750 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-128 Page 127 Peak Area Report Sample Name: 4 x Q C HIGH Plasma Notebook Ratetence: PABX-8VA-1-062 Date Acquired: 10/18/2000 1.54 Operator: MAN Data FileName: 0621038 D Instrument Name: MS7 Data File Path: D:1MSD_0ATA\PAB\PABX-BVA\1-062-n Instrument Vial Number: 39 Instrument Method Name: PA8X_BVA Compound 1) 2) Compound Name MVL MVL-44 Retention Time (Min.) 7 13 7 16 (on 131 135 Peak Area M" 8837 6790 Peak Area Rat 131 6E-2 M* - The lettor "m" in this column indicates that manual integration sms peflormed on that compound. PrintData and Tone: 10/18/2000 12:51 Page 1 000751 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-129 Page 128 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621038.D Vial: 38 18 Oct 2000 1:54 Operator: MAN 4 x QC HIGH Plasma Inst : MSI MS Integration Params : RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) (Abundance ! 5000; 4000! TIC:062138.D A 3000: 2000; 1000; I\ V i-- i ;t . ,-- f 1 I J ' : . . . . . . . . . . . . . . . . . . . . . . . . . . I , -- t - r , I - . . . . . . . . . . . . . . . . . . . . . . . . ' I " ' - ; i - r . i " | . i - i r -- . . . . . . . . . . . . . . ' ' " T ' -- n -- :-- rrin-> 610 6.20 6 30 6 40 6.50 6.60 6.70.6.60 6.90 7 00 7.10 7.20 7,30 7 40 7.50 7-60 7.70 7.60 7,90 MVL Amount : 0.00......... ! Ion: 131 17.133) ............. . j Resp: 8937 I t, 1 /! " ' 7 ' - r-- - - - r - : --------r -- :---- :---r ,T ' .......... ------ I 1 1 1 1 I 1 1 1T ": . : :------------------------^ T "7"' ' H............................ I ' 620 6.30 640 6.50 6.60 6.70 860 6.90 7.00 710 7.20 730 7,40 750 760 7,70 7,80 7.90. IVL-d4________________________________ Amount: 0.00 ' Ion: 135 (7.161) Resp: 6790 A /1 6.20.. 6.30 6.40 B.50 6.60 6.70 6.60 6.90 7.00 7.10 7.20 7 S T `7.40 7 50 7.60 770 760 760 0 6 2 1 0 3 8 . D PABX BVA.M Wed O c t 1 8 1 2 : 5 2 : 0 1 2 0 0 0 Page 2 000752 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-130 Page 129 Peak Area Report Sample Nairn: 4 x Q C MID Plasma Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/18/20002 13 Operator: MAN Data FHe Name: 0621039.D Instrument Name: M S 7 Data FilePath: D:\MSD_DATAIPABiPABX-BVA\1-062-1\ Instrument Vial Number: 39 Instrument Method Name: PABX_BVA Compound 1) 2) Compound Name MVL MVL-04 Retention Time (Min.) 7.13 7.16 tea 131 135 P*ak Area M* 2156 4477 Peak Arsa Rate 481.6E-3 w * The le tte r 'm ' In m il column indicete m et manual integration was performed on that compound. Print Date and Time: 10/18/2000 12:52 Page 000753 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-131 Page 130 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621039.D Vial: 39 18 Oct 2000 2:13 Operator: MAN 4 x QC MID Plasma Inst : MS7 MS Integration Params: RTENT.P Quant Method : D:\HPCHEM\l\MSTHODS\PABX_BVA.M IRTE Integrator) (Abundance 1 TIC: 0621039.D ; 2000 l\ 1 1500 i\ ioooj I sooj I ii i t 11 Ir I7T--''lT :-- I111I--t *- Tim-> 6 10 6 20 6.30 6 <0 6-50 6.60 6 70 6 80 6,90__ 7 00 _ 7.10 _ 7,20_. 7 . 740 7.60 7.60 7.70 7.80 MVL_________ _____________ Amount: _ " 0.00...... l Ion: 131 (7.135) " .. .... .... ...... ........ ! Resp: 2156 7,90........ 6.20 630 6.40 6.50 6 60 670 L-d4.... .... ...... m Ion: 135 (711621 Resp: 4477 I.. r 1'I .I1 6.90 7.00 7.10 7.20 7 30 7JO 7.50 7.60 7.70 7.80 7 90 Amount : . 00 (\ 6.20 6.30 6.40 6.50 6.60 6 70__ 6-60 6 90 7-00 7 10 7.20 7.30 7.40 7 50 7.60 7 70 7.60 7 90 0 6 2 1 0 3 9 .D PABX_BVA.M Wed O c t 18 1 2 : 5 2 : 2 0 2 0 0 0 Page 2 000754 Primedica-Worcester Project number: PABX-BVA 418-018 :PAGE 1-132 Page 131 Peak Area Report Sam ple Name: 4 x QC LOW Plasma Notebook Reference: PABX-8VA-1-062 Dele A cq u ire d : 10/18/2000 2:33 O perator: MAN Data File Name: 0621040.D Data File Patti: D:\MSD_DATA\PABVPABX-8VA\1-Oe2-1\ Instrum ent Method Name: PABX 8VA Instrum ent Name: MS7 Instrum ent Vial Number: 40 Compound # U 2) Comooi^nd Name MVL MVL^J4 R e te n tio n Tim e IMtn.) 7.14 7.16 Ion 131 135 Peak Area M* 1156 5084 Peak Area Ratio 227.4E-3 M* The letter "m" in this column indicates that manual mteoretion was performed on that compound. Print D ate and Time: 10/18/2000 12:52 Page 1 900755 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-133 Page 132 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621040.D Vial: 40 18 Oct 2000 2:33 Operator: MAN 4 x QC LOW Plasma Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) I 2000 1SOO TIC: 0621040.0 MA 1000j /I S0! _ /\ .I I...... .. ,,."i .- F r- IT i t : ,... I, I{. , ., ' ... ..... ...1-', ,-- ime~> 6 10 6.20 6 30 6.40 6.50 6 60 6.70 6.60 6.90 7 00 T IP 7.20 7 30 7.40 7.50 7.60 7.70 7.80 7.90 MVL Amount: 0.00 0 6 2 1 0 4 0 . D PABX BVA.M Wed O c t 18 1 2 : 5 2 : 4 0 2 0 0 0 Page 2 009756 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-134 Page 133 Peak Area Report Sample Name: 4 x Control Plasma Notebook Reference: PABX-6VA-1-0C2 Date A cq u ire d : 10/18/2000 2:52 O perator: MAN Data File Name: D621041.0 Data FMe Path: D:\MSD_DATA\PAB\PABX-BVAM-062-1\ Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Instrum ent Vial N um ber: 41 Comoound # i) 2) Compound Name MVL MVL-44 R e te n tio n Time (Min.) 7 13 7 16 ism 131 135 Peak A rea M* 588 m 4690 Paak Area Ratio 1254E-3 M' e The letter "m" m this column indicates that manual integration was performed on that compound. Print D ate an d Time: 10/18/2000 12:52 Page 1 000757 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-135 Page 134 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621041.D Vial: 41 18 Oct 2000 2:52 Operator: MAN 4 x Control Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) jfcbundanca 200 150oi TIC: 0621041 .D a r j i 1000; II soo; I . , , I ,T".".Y:T-T I',' -T. ' , f'i> ( T - I T ;.. , . ,-i-r ->- . . i , i IT -- -T-| .I ' jrn-> 6.10 6.20 6.30. 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7.30 7 3 7.50 7.60 7.70 7 80 7.90 MVL Amount: 0.00 ; Ion: 131 7.133) ........ ......... ...... iResp: 508 i A .,; ! / / \ 6.20 6 30 6 40 6 50 6 60 6.70 6.80 6 90 7.00 7 10 7 20 7 30 7.40 7 50 7 60 7.70 7 80 7 90 d4 Amount: 0.00 Ion: 135 (7.ISO) Resp: 4690 I\ 620 6.30 6 40 6.50 6 60 6.70 6 80 6.90 7.00 7 10 720 7.30 7.40 7.50 7.60 7.70 7 60 7.90 0 6 2 1 0 4 1 . D PABX BVA .M Wed O c t 18 1 2 : 5 2 : 5 9 2 0 0 0 Page 2 0007541 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-136 Page 135 Peak Area Report Sam ple Name: 5 x QC HIGH Water N otebook Reference: PAflX-BVA-1-062 D ale A cq u ire d : 10/18/2000 3 12 Operator: MAN Data File Name: 0621042.0 Data FMe Path: D:1MSO_DATA\PAB\PASX-8VA\1-062-1V instrum ent Method Name: PABX BVA instrum ent Name: MS7 Inetrum ent Vial N um ber: 42 ComDOund * 1) 2) Com pound Neme MVL MVL-d4 R eten tio n Time (M iai 7.14 7.16 t2B 131 135 Peek A le a M* 7137 5531 Peek Ate Kabo 129 0E-2 M* The letter "m" In this column indicete that manual intagration waa performed cm that compound. Ptint Dete en d Time: 10/18/2000 12.53 Pape 1 000759 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-137 Page 136 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BVA\l-062-l\0621042-D Vial: 42 18 Oct- 2000 3:12 Operator: MAN 5 x QC HIGH Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC 0621G42.D i'\ ; 3Q00| ! ! ; I: iI i! ji 1000) j j i V. ........ Tim-. 6.10 6.20 6 30 6 40 6 50 6.60 6.70 6.60 6 90 7.00 7 10 7.20 7.30 7,40 7.S0 7.60 7.70 7 80 7.90 M V L _____________________________________ Amount : 0.00 _____________ i I o n : 131 (7.137) ; Resp: 7137 620 6.30..... 640 6.50 6 60 S.70 6 80 6.90 7 00 7.10 7 20 7.30 7 40 7.50 7 60 7 70 7.B0 790 0 6 2 1 0 4 2 . D PABX BVA.M Wed O c t I B 1 2 : 5 3 : 1 8 2 0 0 0 Page 2 000766 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-138 Page 137 Peak Area Report Sam ple N n * : S xQC MID Water N otebook Reference: PABX-BVA-1-062 Data A cquired: 10/18/2000 3:31 O p a n to r: MAN Data File Name: 0621043 D Data Fila Path: D:\MSD_DATA\PAB\PA8X-8VAW-062-1V Instrum ent Method Name: PABX BVA Instru m e n t Name: MS7 Instrum ent Vial Number: 43 Comoound # it 2) Compound Name MVL MVL-d4 R e te n tio n DnaiMmJ ten 7 13 131 7.16 135 Peak Area M" 2277 5024 Peak Area Ratio 453.2E-3 W The letter "m" in this column mdlcMee that manual integration was performed on that compound. Print Date and Time: 10/18/2000 12.53 Page 1 000761 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-139 Page 138 Quantitation Report (QT Reviewed) Data File Acq On Sample Mbc D:\MSD_DATA\PAH\PABX-BVA\1-062-1\0621043.D Vial: 43 18 0ct~2000 3:31 Operator: MAN S x QC MID Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundance ....... ... TIC. 0621043 0 i 2000; !1 !i oooi i I / / L T- I 7 , T-- I I ' ' ' I 1 I ' T ' ' ' 1 r'-l . " > >-- - n 1 1 - n -- ! ' T l I-- Time- 6 10 6.20 6.30 640 6.50 6.60 670 6.B0 6 90 7.00 7 10 7 20 7 30 7 40 750 7 60 7 70 760 7S0 MVL _______________________ Amount :_______0.00_________ Ion: 131 (7.134) " ... ......~ ! Resp: 2277 ;i i! I\ ! 6 20 6.30 J5.40 6,50, 6 60 6.70 6.80 630 7.D0 7 10 7 20 7.30 7.40 7.S0 7.B0 7.70 7 . 750 'L -d~4_______ _________ Amount :_______0.00_______ Ion: 135 (7 . T i ) ....... . . ...................... j Resp: 5024 i 1'I'' 6.20 6.30 640 6.50 6.60 6.70 6.80 6.90 7 00 7 10 7.20 7.30 7 40 7 50 7.60 7.70 7 80 7.90 0 6 2 1 0 4 3 .D PABX_BVA. M Wed O ct 18 1 2 : 5 3 : 3 8 2 000 Page 2 000762 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-140 Page 139 Peak Area Report Sample Nam: 5 x QC LOW Water N otebook Reference: PABX-BVA-1-062 Dale A cq u ire d : 10/1B/2000 3:50 O perator: MAN Data File Name: 0621044.D Data File Path: D:\MSD_0ATA\PAB\PABX-BVA\i -062-1\ Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Instrum ent Vial N um ber: 44 ComDOund # 1) 2) Comoound Nam* MVL MVL-d4 R eten tio n Time (Min i 7.13 7.16 ion 131 135 Peak Area M* 665 4124 Peak Area Ratio 168.5E-3 M* * The letter "fn" in Itnx column indicele that manual mleyreoon was performed on that compound P nm D ate and Tima 10/18/2000 12:53 Page 1 0007G 3 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-141 Page 140 Quantitation Report (QT Reviewed) Data File Acq On Sample M BC D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621044.D Vial: 44 18 Oct 2000 3:50 Operator: MAN 5 x QC LOW Water Inet : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance j 2000 TIC: 0621044.0" A 1500 lOOOj soo| I r ,=n,4- r".7 I: \~T, --T' TI !\ i\ 'j ' ' ' 'H' '""'''T '"'--' ! ' T" 1! ' ''lT' : - ' 1t't" --r'r*T~' " Time--> 6.10 6.20 6.30 640 6.50 6 60 670 6 B0 6.90 7 00 710 7.20 7.30 7.-40 7SO 7,60 7.70 7.60 7 90 MVL Amount : 0.00 0 6 2 1 0 4 4 .D PABX BVA.M Wed O c t 18 1 2 : 5 3 : 5 7 2000 Page 2 Primedica-Worcester Project number: PABX-BVA 418-018 .-PAGE 1-142 P age 141 Peak Area Report Sample Name: 5 QC LLOQ Water Notebook Reference: PAfiX-BVA-1-062 Date A cquired: 10/18/2000 4 10 O perator: MAN Date File Name: 0021045.0 Instrum ent Name: MS7 Date FMo Path: D: VMSDJOATAiPABVPAaX-BVAVI -062-1V Instrum ent Vial N um ber: 45 Instrum ent Method Name: PA8X_BVA Compound # 1) 2) Compound Name MVl MVL-d4 R e te n tio n Time Min.l 7.14 7.16 ton 131 135 B tA fe ftrg j eai 314 4172 752.8E-4 M- The letter `rvT in ttiii column indicate mt manual integration wea performed on that compound. Print Date and Time: 10/1&/2000 12:54 Page 000765 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-143 Page 142 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621045.D Vial: 45 18 Oct 2000 4:10 Operator: MAN 5 x QC LLOQ Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundare j 2000i TIC: 062104S.D ! ! 1500J |i j 1000; j 500 iU Jv1i l ! Tima-* 6.10 6J20 6.30 640 6.S0 6.60 670 6.B0 6.90 7.00 710 7.20 730 7.40 7.50 7.60 7.70 7.B0 7.90 MVL Amount : 0.00 Ion: 131 (7.138) ' R esp: 314 I ___ _ ______ . J Al _ I 6.20 6.30 6.40 6 50 6.60 670 6.80 6.90 700 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 ^^IVL-d4 Amount : 0.00 TM Ion: 135 (7.162) ! R e s p : 4172 ft I -, I-,-', , .- m-.- r , ......... r r-----"'i-' . . . . ; ...........................................................................T '-- ........... r -.-r'i ......................................rrr| ! ! 6.20 6.30 6.40 6.50 660.... 6.70 6.B0 6.90 7 00 7.10 7.20 7.30 7.40 7.50 7.60.... 7.70 7JO. ...7JO........ I 0 6 2 1 0 4 5 .D PABX_BVA.M Wed O c t 18 1 2 : 5 4 : 1 7 2 0 0 0 Page 2 00076 Primedica-Worcester Project number: PABX-BVA 418-018 :PAGE 1-144 Page 143 Peak Area Report Sam pia Name: 5 x QC HIGH Plasma N otebook Reference: PABX-BVA-1-062 Date A cquired: 10/18/2000 4:29 O perator. MAN Data FU* Name: 0621048.D Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1-062-1\ Insbum ent Method Name: PABX-BVA In stru m e n t Name: MS7 In stru m e n t V ial N um ber: 48 Com bound t V 21 gWMHfflPUJyanw MVL. MVL-d4 R e te n tio n liaw-iMiuJ 7 13 7.18 Isn 131 135 P e a k A te a M* 7218 5671 Peak Area Ratio 127.3E-2 M* The lattar "m" in tuia coturni indicates that manual m ta p ta ta waa pertormad on that compound. Pnnt D ata and Time: 10/18/2000 12:54 Page 1 000767 Primedica-Worcester Project number: PABX-BVA 418-018.-PAGE 1-145 Page 144 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621046.D Vial: 46 18 Oct 2000 4:29 Operator: MAN 5 x QC HIGH Plasma Inat : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) burntTM* .......... i 4000; 30ooiI I 2000;i 1000; " .............. ~' j \TIC: 0621046.0 !!'; I! \ !\ .Tune- 6.10 6,20 6.30 6<0 6.60 .fee ' 6.7Q 6.60 " e . - - 7.00 ' 7~10 A20 ' 7 30 740 7 50 7 60* 770 T.B0 7.90 MVL Amount : 0.00 0 6 2 1 0 4 6 . D PABX BVA.M Wed O c t 18 1 2 : 5 4 : 3 6 2 0 0 0 Page 2 00096ft Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-146 Page 145 Peak Area Report Sample Name: 5 x QC MID Plasma Notebook Reference: PABX-8VA-1 -062 Date A c q u ire d : 10/10/2000 4:49 O perator: MAN Date File Name: 0621047.D Date Fite Path: D.\M$0_DATA\PAB\PABX-BVA\1<0621\ Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Inetrument Vial N um ber: 47 Compound # U 2) C om pound Napie MVL M Vl-d4 R eten tio n T im * (Min.) 7 14 7.ie log 131 135 Peak Area W 2502 5132 Peak Ansa Ratio 4B7.5E-3 M" The letter "m" in tht column indicele thel menuel in tag ra S o n wee p erform ed on that com pound. Print D ata an d Time: 10/16/2000 12:54 Page 1 000769 Priniedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-147 Page 146 Q u a n tita tio n R eport (QT R e v ie w e d ) D ata P ile Acq On Sam ple M ise D:\M SD__D ATA \PA B\PABX -BVA \l-062-l\0621047.D V ia l: 47 18 O ct 2000 4 :4 9 O p e r a t o r : MAN S x QC MID P la s m a In st : MS7 MS I n t e g r a t i o n P a r a m s : R T E IN T .P Q u a n t M e th o d : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE I n t e g r a t o r ) ^buodance ii ' 2000! TIC: 0621047.D t l\ '1 1 ! 10001 ;| !i !/ \\ Tn-> 6.10 6 20 6.30 6.40 6.50 6.60 6.70 6.60 6.90 7 00 7.10 7.20 7 30 7,40 7.50 7.60 7 70 7.B0 7.90 MVL Am ount: 0 .0 0 Io n : 131 (7.136) R esp: 2502 : i\ ' 6.20__ 6 30 6.40 650 6.60 6 70 6.60 8.90 7.00 7.10 720 7,30 7 40 7.50 7 60 7 70 7 60 7 90 -d 4 Amount : 0.00 " ' yI o n : 135 ( 7 .1 6 0 ) R esp: 5132 A I\ I 6.20 6.30 6 40 6.50 6.60 6 70 6 60 6.90 7 00 7,10 7 20 7.M 7.40 7.50 7.60 7.70 7-BO 7.90 0621047.D PABX_BVA.M Wed Oct 18 12:54:55 2000 Page 2 000779 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-148 Page 147 Peak Area Report Sample Neme: 5 x QC LOW Ptaema N otebook Reference: PABX-BVA-1-062 Data A cq u ire d : 10/18/2000 5:08 O p e ra to r MAN Data File Name: 0621048.0 Data File Path: D:1MSD_DATA\PAB\PABX-BVA\1 -062-H Instru m e n t Method Name: PABX BVA In stru m e n t Name: MS7 Instrum ent V ial N um ber: 48 ComDound # 1) 2) Compound Nrrw MVL MVL-d4 R e te n tio n Tim stlm J 7.13 7.16 te " 131 135 PuaK Area M* 744 3350 Pxfc A m a R a t 222.1E-3 M* Th* letter "m" in this column Indice! that manual integration we* performed on the! compound Print Oate and Time-10/18/2000 12:55 Page 1 000771 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-149 Page 148 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621048.D Vial: 48 18 Oct 2000 5:08 Operator: MAN 5 x QC LOW Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M IRTE Integrator) pJwxtanoT....." i 1500; : ............. TIC 0621048 D fi I ......................... 1000! I! !I 500 !\ / !' Time-> 6.10 620 6 .30 6.40 .... ! V .'i-ri-P 'r-i-i-ff-i-J-FTa i'i .'T'-."r'....... - r : .'T '---- 1 . i-i-ri-- "| I 6.50 6.60 6.70 _6.80 6.90 7.00 7.10 7.20 7.30 7 40 7.50 7.60 7.70 | '. 7-B0 7,90 MVL " ~ A m ount: 0.00 0621048.D PABX_BVA.M Wed Oct 18 12:55:14 2000 Page 2 000772 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-150 Page 149 Peak Area Report Sam ple Name: 5 xControl P lu m s Notebook Reference: PASX-BVA-1-062 Data A cquired: 10/18/2000 5:28 Operator: MAN Data File Name: 0621Q49.D Data File Patti: O:\MSD_OATA\PA8\PABX-eVA\1 -062-1\ Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Instru m e n t Vial Number: 49 Comoound # 1) 2) Comoound Name MVl MVL-cW R e te n tio n Time fMirvi 7.13 7.16 las 131 135 Peak Area M* 396 3344 Peak Area Rabo 118.4E-3 U - * Tha teller V m this cotumn mdicalee that manual integration eras performed on that compound. Pnnt Date and Time 10/18/2000 12:55 Pttqe 1 000773 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-151 Page 150 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-0S2-1\0621049.D Vial: 49 18 Oct 2000 5:28 Operator: MAN 5 x Control Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) ADunaanc*.... : 1I 1S00 i! ;> ; 1000! '~ TC. 0621040.0 I)I I ... 6-20 6.30 6.40 6.50 6.60 6-70 6.60 6.90 7.00 7.10 JT.2D 7.30 7.40 7.50 7.60 7.70 7.80 7JO 0621049.D PABX BVA.M Wed Oct 18 12:55:34 2000 Page 2 000774 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-152 Page 151 Peak Area Report S a m p * Nam: 6 QC HIGH W ater Notebook Rateranca: PABX-BVA-1-082 Data A cquired: 10/18/2000 5 47 O p e ra to r MAN Data FHe Name: 06210S0.D Data F I* Patti: D:\MS0_DATA\PAB\PABX-8VA\1-062-1 1 Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Instrum ent V ial Num ber: 50 Congrui * 1) 2) Compound Name MVL M V l-d4 R eten tio n Tim e (Min.) 7.14 7.17 iss 131 135 Peak Area M* 7184 5456 Peak Area Ratto 131.6E-2 M* The letter "m" in thia column inctoceh* thet manual integration was partomad on that compound. Print Date and Time: 10/18/2000 12:55 Page 1 000775 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-153 Page 152 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621050.D Vial: 50 18 Oct 2000 5:47 Operator: MAN 6 x QC HIGH Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance 4000; TIC: 06210S0.D |\ 3000i lj 1 200oj ! 1000-' i i r rr*me~> 6.10 6.20 6.30 6.40 6 50 MVL i I o n : 131 (7.138) I Reap: 7184 6.80 6.70 6.80 6.90 (: I\ !y 7.0 7 10 7.20 Amount : 0.00 l \ 20 6.30 6.40 6.50 6.S0 -d4 ilon: 135 (7.166) \ Resp: 5458 6.70 6 90 6 90 7.00 __ 710 7.20 7.30 Amount: 7.40 7,50 0.00 7.60 ......7.70 7.60 7.90 --n-- ' I'` 7.806.20 6,30 6.40 6.50 5.60 6 70 5.80 690 700 7.10 7.20 7.X 7 40 7JO 7.60 770 7,90 0621050.D PABX BVA.M Wed Oct 18 12:55:53 2000 Page 2 000776 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-154 Page 153 Peak Area Report S am ple Mama: 6 x QC MID Water Notebook Reference: PABX-8VA-1-062 Date A cq u ire d : 10/18/2000 6:07 Operator. MAN Data File Name: 0621051 0 Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1-062-1\ Inatrument Method Name: PA8X_BVA Inatrum ent Name: MS7 In a tru m e nt Vial Num ber. 51 ComDound # 1) 2) ComDound Name MVL MVL-dd R eten tio n Time IMIn.l 7.13 7 16 22 131 135 Peak Aree M* 2958 m 6314 Peak Area Ratio 468.5E-3 W The letter "m" In ime column indicate that manual integration wee pertormad on that compound Print Date and Time; 10/1B/2000 12:56 Page i 000777 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-155 Page 154 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621051.D Vial: 51 18 Oct 2000 6:07 Operator: MAN 6 x QC MID Water Inst : MS7 MS Integration Pararne: RTEINT.F Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance j | 3000j ! 2000j 1000 TIC: 0621051.D /, A I I I----- --- ,,r ,, F.'-'iif ,-rTi'W T ii.. -- '-' 6.10 6.20 6.30 6 40 6.50 660 6.70 6.80 6.90 7.00 7,10 7.20 .. 7,30 7.40 7,50 7 60 7 70 7.80 7.90 MVL _____ _______________________ Amount : ___ 0 -00_______________ j IonY 131 (7.134) ...... j Reap: 2958 .20 6,30 6.40 6.50 6.60 -d4 Ion: 135 (7.162! ;Resp: 6314 6.70 6 60 . 6.90.....7,00.......7.10 7.20 7 30 ________ Amount:______ A l\ 7.40 7.50 0.00 7.60 7 70 7 60 7.90 ! 6.20 6.3 6.40 6.506.60 6.70 6.60 6.9Q 7JO 7.10 J I 1 '' I 1 * 1 1 , " T " ' 1- " ' ' r ' ' T ' 1 ' 1 7.20. 7.30 7 40 7.50 7.60 7,70 7.B0 790 -t 0621051.D PABX_BVA.M Wed Oct 18 12:56:12 2000 Page 2 000778 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-156 Page 155 Peak Area Report Sample Name: 6 x QC LOW Water N otebook Reference: PABX-BVA-1-062 Date A cquired: 10/18/2000 6:26 Operator: MAN Data File Name: 0621052.0 Data File Path: O:\MSD_DATA\PAB1PABX-BVAVI-062-1\ Inatrum ent Method Name: PABX_BVA Inatrument Name: MS7 Inatrum ent Vial Number: 52 Comoound # 1) 2) Compound Name MVL MVL-CJ4 R e te n tio n T im e ton Peak Aree M* Peak Area Ratio 7.13 131 1038 167.9E-3 7.16 135 61B7 M* Th lattar V In mis column indicates that manual Integralien was pertormad on that compound. Print Date and Time: 10/18/2000 12:56 Pepe 1 000779 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-157 Page 156 Quantitation Report (QT Reviewed) Data File Acq On Sample Miac D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621052.D Vial: 52 18 Oct 2000 6:26 Operator: MAN 6 x QC LOW water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abunane* ! 30001 TIC: QS21QS2.0 j) !I ;; ` li Il S 2000! ji li h rnt! /1 ir rrime-> 6.10 MVL ....... 6.20 6.30 11 T*1 T-I T I 1 r n ' i I I I F | . V . ' .T ' 1' I l r ^ , I . I " '- n --I-- I V - ' 1 I-- I . r -r - ~ T T ! - | -- I I . 6 40 6 50 6.60 6 70 6 80 6 90 7.00 7 10 7 20 7.30 7 40 7 50 7.60 7 70 7.60 7 90 ... ~ "Amount: 0.00 [ j 0621052.D PABX BVA.M Wed Oct 18 12:56:32 2000 Page 2 000780 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-158 Page 157 Peak Area Report Sam ple Name: 6 X QC LLOQ Water N otebook Reference: PABX-BVA-1-062 Date A cq u ire d : 10/18/2000 6:46 O perator: MAN Data File Name: 0621053 D Data File Path: D:\MSD_DATA\PAB\PABX-BVA\1-062-1l Instrument Method Name: PA0X_BVA Inatrum ant Name: MS7 In stru m e n t Vial N um ber: 53 Compound 1| 2) Compound Name MVL M V l-d4 R e te n tio n Tim e (Min.) 7.14 7.16 Ion 131 13S Peak At m M* 462 m 6048 Peak Area Rabo 7639E-4 W The letter "m" tn tht* column tndtcetee thet menuet Integration wee performed on that compound Print Date and Time: 10/18/2000 12:56 Page 1 000781 Primedica-Worcester Project number: PABX-BVA 418-018 :PAGE 1-159 Page 158 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621053.D Vial: 53 18 Oct-2000 6:46 Operator: MAN 6 x QC LLOQ Water Inst : MS7 MS integration Params: RTEINT.P Quant Method : D:\HPCKEM\1\METH0DS\PASX_BVA.M (RTE Integrator) Abundan 3000I TIC: 0621053.D 2X0 I ^ --n-- .-*; '..... . r, r, ,.. --| ..... .-r-- --- -- i- Tim- 6.10 6 20 8.30 6.40 6.50 6.60 6.70 6.80 6.30 TOO 7,10 7.20 7.30 7.40 7.80 7 60 7.70 7.80 7-90 MVL Amount: 0.00 0621053.D PABX BVA.M Wed Oct 18 12:56:51 2000 Page 2 000782 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-160 Page 159 Peak Area Report S am ple Nam: S X OC HIGH Plasma Notebook Reterence: PABX-BVA-1-062 Date A c q u im i: 10/18/2000 7:05 Operator: MAN Date Fite Name: 0621054.D Date Fite Path: O:\MSO_DATA\PABV3ABX-BVAU-062- 1\ Instrum ent Method Name: PABX_BVA In stru m e n t Name: MS7 Instru m e n t V ial Num ber: 54 Compound # i 2) Compound Name MVL MVL.-44 R e te n tio n Ito iM lB J 7.13 7 18 Ion 131 135 Peak Area M* 4406 3506 Peak Area Ratio 125.8E-2 HT The letter Hm** In Itila column indicatea that manual integration wae performed on that compound. Print Date and Tima: 10/18/2000 12.57 Pag* i 000783 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-161 Page 160 Quantitation Report (QT Reviewed) Data File Acq On Sample MiSC D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621054.D Vial: 54 18 Oct 2000 7:05 Operator: MAN 6 x QC HIGH Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) K&urxtenct ; 25coi ;2000 TTC: 0621054D isooj 1000 i \900 rrrr.-q rv T p p iTim-> S to 6 20 6 30 6 40 6 90 6 60 6.70 680 6 90 7.00 710 7 2C 7.30 7.40 7 50 MVL _________ ____________ ' Amount :.... 0.00 Ion: 131 (7.132) Resp: 4406 7.60 7.70 7.90 7 90 ___________ \ 6.20 6.30 6 40 6.50 6 60 L-d4 Ion: 135 (7.159) i Resp: 3508 6.70 6.60 660 7 00 7.10 7.20 7.30 Amount : 7.40 7.50 0.00 7.60 7.70 7 80 . 7SO . /V ! 6.20 6.30 . 6.40 650 6.60 6 70 6.60 6.90 7.00 7.10 7.20 7.30 7 40 7.90 7.60 7.70 7J"~^7.S0 ' 0621054.D PABX BVA.M Wed Oct 18 12:57:10 2000 Page 2 000784 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-162 Page 161 Peak Area Report Sample Name: 6 x QC MID Plasma N otebook Reference: PABX-BVA-1-062 Date Acquired: 10/18/2000 7 25 Operator: MAN Date Fite Name: 0621055.D Date File Path: 0 \MSD_0ATA\PAB\PABX-BVAVI-0e2-1\ Instrum ent Method Name: PABX_8VA Instrum ent Name: MS7 Instrum ent Vial Number: 55 Comoound # 1) 2) Comoound Name MVL MVL-d4 R e te n tio n Tim e (Min.) 7 13 7 18 ion 131 135 P ik A a s m* 2452 4988 Peak A rea Ratio 4916E-3 M* * The letter *m" in this column indicates that manual integration was performed on that compound. Print Dale and Time 10/1B/2000 12:57 Page 1 00078S Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-163 Page 162 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621055.D Vial: 55 18 Oct 2000 7:25 Operator: MAN 6 x QC MID Plasma Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) jA&undanee i 2000i; 1000: TIC: 0621065 D r, i/!; / i ! i\ i; /\ J.;: ..1r ,-T-,.I-11,..w 'i"T1 -*-""?r^ m\... . 1, -r Time-. 6.10 6.20 6.30 6.40 6.50 6.60 6.70 6 80 6.90 7.00 7.1D 7.20 7.30 7.40 7.50 7.60 7.70 7.60 7 S0 MVL Amount : 0.00 ; Ion: 131 (7.133) i Resp: 2452 i ! j A A IV r-'rr~r'[ p ~i1 ^mVL,;d4_ _` 6.20 6.X 6.40 6,50 6 6 0 6.70 6.BO 6 9 0 7 00 7.10 T.2B T . 7.40 7.50 7.60 7.70 7 .B0 78 0 Amount : 0.00 ^ " T o n : 135 (7 161) i Resp: 4988 I/__\v i 6.BO"r `T` r ` , r-r "v'11 I ............... r-' -P- 1 i 6.20 6.30 6.40 6.50 6.60 . 6 70 6 9 0 7.00 7.1 Q 7,20 7.30 7,40 7.k> 7.60 7.70 r-rT ... '.B0 7.90 0621055.D PABX BVA.M Wed Oct 18 12;57;30 2000 Page 2 00078 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-164 Page 163 Peak Area Report Sample Name: 6 * Q C L O W Ptaema Notebook Reference: PABX-aVAr1-062 Date Acquired: 10/18/2000 7 44 Operator: MAN Data File Name: 0621056.D Instrument Name: MS7 Data File Path: 0:1MSD_DATA\PA81PABX-BVA\1-062-11 Instrumen Vial Number: 58 instrument Method Name: PABX_BVA Compound 1) 2) Comoound Name MVL MVL-44 Retention Tene (Mln.1 7.14 7.16 ten 131 135 PitKftm m : 1178 5138 Peak Ares Ratio 2289E-3 IT Th toit*1**m" m th is colurrm indicato ttwii m anual in to n a tio n w a s perfo rm ed oft th a t com pound. Pim t Date and Tim e: 10/18/2000 12:57 Page 1 000787 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-165 Page 164 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621056.D Vial: 56 18 Oct 2000 7:44 Operator: MAN 6 x QC LOW Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundanc* ! 25o' j 2000j ! 1500 TIC: 0621066.D `\ Ij I! iT w -> 6.10 6.20 6.30 6.40 6.60 6 60 6 70 6 80 ~ 6.90 7.00 7 10 7.20 7 30 7.40 '7.50 ' `7 6 0 7 70 7 80 7.90 MVL " Amount: 0.00 0621056.D PABX BVA.M Wed Oct 10 12:57:49 2000 Page 2 000788 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-166 Page 165 Peak Area Report Sample Name: 6 x Control Plasma Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/16/2000 8 04 Operator M A N Data FHe Name: 0621Q57.D Instrument Name: MS7 Data File Path: D:\MSD_DATAVPABVPABX-BVAt1-082-n InstrumentVialNumber: 57 Instrument Method Name: PABX_BVA Compound * U 2) Comoound Nam* MVL MVL-d* Retention Time IMtn.i 7.13 7.16 ten 131 135 E M K A m M* 503 4373 Reek Are Recio 115.0E-3 M* * Ths letterV inthiscolumn indlcaleethatmanuel Integrationwee penormad an Ihetcompound. Print O sle and Time: 10/18/2000 12:58 Pag* 1 000789 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-167 Page 166 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-0S2-1\0621057.D Vial: 57 18 Oct 2000 8:04 Operator: MAN 6 x Control Plasma Inst : MS7 MS Integration Params : RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) ^Aundsoce i 200o! 1500- TIC: 0621057.0 ! 1000 500 fTime-> 6.10 520 6 30 6 40 6.50 6.60 6.70 6.80 JS.90 MVL.... ________________________ ..Ion:"131 (7.134) Resp: 503 1' ,. ^ , ^ r ^ ,^ rr^ r T T r ,. ... , n ...----- ; - TT-. 7.00 7.10 7,20 7.30 7.40 7.50 7 60 7.70 7.B0 7,90 Amount: 0.00 ____ _____ I )V T1 r-r .> 6 2 0 6,30 6 40 6 50 6 60 6 70 6.80 6 90 7 00 7 10 7.20 7.30 7 40 7.50 7.60 7 70 7,60 7.90 m L-d4 Ion: 135 (7.162) Amount : 0.00 ! Resp: 4373 I 6.20 6.30 6.40 6 50 6.60 6.70 6.60 6 90 7 00 7.10 7 20 7.30 7.40 7.50 7.60 7.70....7.80 7 90____j 0621057.D PABX BVA.M Wed Oct 18 12:58:08 2000 Page 2 000790 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-168 Page 167 Peak Area Report Sample Mame; 2 * BlankWalet Notebook Reterence: PABX-BVA-1-062 Date Acquired. 10/18/2000 8:23 Operator: M A N Data File Name: 0621058 0 Instrument Name: MS7 Data FilePath: D:\MSD_DATA\PAB\PABX-8VA\1-0e2-1\ Instrument VialNumber: 58 Instrument Method Name: PABX BVA Compound # 1) 2) Compound Name MVL MVL-d4 Retention Time IMIn.l 0.00 000 ton 131 135 Peak Area M* 0 0 Peak Area Baflp 3IV/0! r r The tetter I n " m th is cottane indicates that m anual integration w aa perterm ad on that componard. P rint Dale and Tima: 10/18/2000 12:58 Page 1 000731 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-169 Page 168 Quantitation Report (QT Reviewed) Data File Acq On Sample Miac D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621058.D Vial: 50 18 Oct 2000 8:23 Operator: MAN 2 x Blank Water Inst : MS7 MS Integration Parama: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M {RTE Integrator) yhbundanc* TIC: 0621058.0 A 250 / \ 200-: A |TTM-> 6 10 6.20 6 30 6.40 6.50 6.60 6.70 6.80 690 7,00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 MVL _____ Amount : Not Found____________________ Ion: 131.(O'.000) i\ !1 '-\S\ r V.,. j 1 1 ' ' ' ---------------------- : I m r1'""' t T t '-- ' I ' ' I ' ----- ------------------ -"I"' ' > I---- t v ... 6.20 6 30 6.40 6.50 6.60 6.70 6.80 6 90 7 00.. 7.10 7.20 7.30 7 40 7.50 7.60 7.70 7.80 7 90 '7L-d4 Ion: 135 (0.000) ________ * Amount: Not Found'....... /: 6 20 6 30 .6,40 6 50 6.60 6 70 6.60 6.90 7 00 7 10 7 20 7.30 7 40 7.50 7.60 7.70 7.80 700 0621058.D PABX_BVA.M Wed Oct 18 12:58:28 2000 Page 2 000792 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-170 Page 169 Peak Area Report Sample Name: 2 x Contro*Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/18/2000 8 43 Operator: M A N Data File Name: 0621059. Initniment Name: MS7 Data File Path: D:\MS0_DATA\PAB\PABX-8VA\1-062-1\ Instrument Vial Number: 59 Instrument Method Name: PASX_8VA Compound 1) 2) Compound Name MVL MVl-d4 Retention Time IMin.l 0.00 7.16 Ion 131 135 Peek Aree M* 0 5369 Peak Area Rath 000.0E-3 MT Tho le tte r Hm " in th is colum n in d icates tt>at m enu*! integration mm porform od cm th at com pound. Print Data and Time: 10/18/2000 12:56 Pag 000733 Primedica-Worcester Project number: PABX-BVA 418~018:PAGE 1-171 Page 170 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSDJDATA\PAB\PABX-BVA\l-062-l\0621059.D Vial: 59 18 Oct 2000 8:43 Operator: MAN 2 x Control Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\h p c h e m \i \m e t h o d s \p a b x _b v a .m (RTE Integrator) lihunclanca TIC:06210S9.D 20 jiI i'r~~.VT-T' ' 1:' 1' I11 !11' 1I11' 1r "''I1*.''TTl' I' ' 'I ' I'i'll-.1....">.......... .;--1.. --'"T'*""' fTime-> 6 10 6.20 6.30 6 40 6.80 6 60 870 8 80 690 7.00 710 7 20 7. 7.40 7.50 7 60 7 70 7.60 7 90 MVL Amount: Not Found.... Ion: 131 (0.000) X~A~'vv/ v / \ __ ;"~n-- '11!'1i!' 11i 1111`1 1 6-20 6.30 6 40 6.50 6.60 670 MVL-d4 W Ion: 135 (7.163) : Resp: 5369 ...... 6.80 6.90 7.00 i i I | ! 6.20 6.30 6.40 6.60 6.60 6.70 680 6.90 700 7.10 7.20 7.30 Amount : A a 7.10 7.20 7.30 7 40 7.50 0.00 7.40 7.ko A -/A -- r-r-T-- ro 7.70 7.80 790 7.60 7.70 7.BO 7.80 0621059.D PABX BVA.M Wed Oct 18 12:50:47 2000 Page 2 000794 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-172 Page 171 Peak Area Report Sample Name: 2 xSTD-1 Water Notebook Reference: PABX-8VA-1-062 Date Acquired: 10/18/2000 9 02 Operator MAN Data File Name: 0621060.D Instrument Name: MS7 Data FitePatti:D:\MSD_0ATA\PABIPABX-BVA\1-062-1\ Instrument Vial Number. SO instrument Method Name: PABX_BVA ComDound # U 2) Comootmd Name MVL MVL-dd Retention Time (Min.) 7.13 7 16 Ion 131 135 Peak Area 487 6305 Peak Area Ratio 772.4E-4 M" * T he letter "m" in thie colum n in d ic a te s th a t m anual integration w a s perform ed on th a t compound. P rint Date and Time: 10/18/2000 12:58 Page 1 000795 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-173 Page 172 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621060.D Vial: 60 18 Oct 2000 9:02 Operator: MAN 2 x STD-l Water Inst : MS7 MS Integration Parama : RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) 0621060.D PABX BVA.M Wed Oct 18 12:59:06 2000 Page 2 000796 Pnmedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-174 Page 173 Peak Area Report Sample Name: 2 x STD-2 Water Notebook Reference: PABX-6VA-1-062 Date Acquired: 10/18/2000 9:22 Operator: M A N Data FileName: 0621061.D Instrument Name: MS7 Data File Path: 0:1MSD_DATA\PAB\PABX-BVAV1-062-1\ Instrument Vial Number: 61 Instrument Method Name: PABX_BVA Compound 1) 2) Compound Name M V L MVL-ctf Retention Time fMin.) 7U 7.16 jOQ 131 135 Peat Area M* 781 m 5494 Peak Area Ratio 142.2E-3 M* T he loHor "m " in Mum co lu m n in d icated th at m anual integration w aa p a rto tm ad on th at com pound. P rint Date and Time: 10/18/2000 12:59 Paga 1 000797 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-175 Page 174 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621061.D Vial: 61 18 Oct 2000 9:22 Operator: MAN 2 x STD-2 Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HFCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC: 0621061.0 ! I2000j I! I 1000i T ',-- T' ~ ' 'I" *1" 1 ' r r ^!T- 6.10 6-20 6.30 6.40 6.50 6.60 6,70 MVL_____________________ T Ion: 131 (7.137) I Resp: 7B1 V TT ' V . - I I ! t ,"l , ,1"rj . . I , I T n --T . T p -- I ! I . , , I I I , 6.90 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 ~Amount :_______0.00 *" A l~r 7.80 7 90 ______ I\ i 6.20__ 6 30 6. 6.50 6.60 6 70 6.B0 6.90 7.00 7.10 7.20 7.30 740 7.50. 7 60 7.70 7.80 790 d4 Amount : 0.00 Ion: 135 (7.160) ; Resp: 5494 /\ I' 'r-. T , . 'v^rrrrrn 1I1111I1 '-pr-r 6.20 6.X 6.40 6.50 6.60 6.70 6-80 630 7.00 710 720 7.30 740 7.50 7.60 7.70 7.90 7.90 0621061.D PABX BVA.M Wed Oct 18 12:59:26 2000 Page 2 000799 Primedica-Worcester Project number: PABX-BVA 418-018 :PAGE 1-176 Page 175 Peak Area Report Sample Name: 2 x STD-3 Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/18/2000 941 Operator: MAN Oats PileName: 0021062.D Instrument Name: MS7 Data FilePath: O:\MSO_OATA\PAB1PABX-8VAM<062-1\ Inatrument VialNumber 62 instrument Method Name: PAJBX_BVA Compound 1) 2) Compound N w MVL MVL-04 Ratontlon Time |M)n,| 7 14 7.16 Isffl 131 135 tc 1305 6271 Ar?t Ratta 208.1E-3 M* T ha lattar " m " in this colum n indicata m at m anual in ta g ralio n warn parto rm ad o n tttat com pound. P rint Date end Tim e 10/18/2000 12 50 Page 1 000799 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-177 Page 176 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621062.D Vial: 62 18 Oct- 2000 9:41 Operator: MAN 2 x STD-3 Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHKM\1\METH0DS\PABX_BVA.M (RTE Integrator) iMDundanos j 3000! 2000i fC0621062.0 n i; /I I t**-- . 1-- I ' I ............... ' I -- ' 1 i - - - I 1 """1 ' ' '-- r ' " T - " ' I1'' " ' ' ' 1 I-- 1 1 ' I '-- -- i-- ' T -- ' I " " ] I [Tw k -> 6 10 6 20 6.30 640 6 60 6 60 670 6 90 6.90 7.00 710 7.20 7.30 7.40 7.50 7.60 770 7 80 7.90____ | MVL ........ * ______________________ A mount:______ 0 .00______________________ [ Ion; 131 (7.135! ; :! Resp: 1305 1 n ;i , ; . .. 'I 6.20 6.30 6.40 6.50 6.60 -d4 Ion: 135 (7.162) ! RDea sp: 6271 J v. .................... 6,70 6.80__6.90 7 00 710 7.20 7.30 Amount: l\ f 740 7.50 0.00 7.90 . - y . -r 7.7P 7.80 7.90 t I 620 6.30 6.40 6 50 6.60 6.70 660 6.90 7.00 7,10 7.20 7.X 740 7.50 760 770 7.80 7.90 0621062.D PABX BVA.M Wed Oct 18 12:59:47 2000 Page 2 000600 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-178 Page 177 Peak Area Report Sample Name: 2 x STD-4 Water Notebook Reference: PASX-BVA-1-062 Date Acquired: 10/18/2000 10:01 Operator M A N Data Fite Name: 0821063.D Instrument Name: MS7 Data FilePath: D:IMSD_DA7Atf>AB\PA8X-8VAt1-062-1\ InstrumentVialNumber: 83 Instrument Method Name: PABX_BVA Compound * 21)| Compound Name MVL MVL-d4 Retention Time (Min.) 7.13 7 16 Isa 131 135 Peek Area M* 1917 m 5685 Peak Araa Rat 337.2E-3 W * The letter V in thie colum n indicatet that m anual integration was perform ed on that compound P rint Date and Time: 10/18/2000 12:59 Page t 000S01 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-179 Page 178 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSDJ3ATA\PAB\PABX-BVA\1-062-1\0621063.D Vial: S3 18 0ct~2000 10:01 Operator: MAN 2 x STD-4 Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M IRTE Integrator) Itbundanos 3000; j 2000! ;1 1000 TIC: 0621063.0 i\ !i a !| /! ;\ /V hT-> 610 6.20 6.30 640 6.50 6.60 6.70 6.60 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.60 730 MVL Amount: 0.00 | Ion: 131 (7.134) 1 Resp: 1917 A ' 6.20 6.30 6 40 6.50 6.60 6 70 6.80 6 90 7.00 7 10 7.20 7.30 7.40 7.50 7 60 7.70 7.60 7.90 /L-d4 Amount: 0.00 Ion: 135 (7.161) Resp: 5685 /\ 6.20 6 30 6.40 6.50 6.60 6.70 6.B0 6 90 7 00 7.10 7.20 7.30 7.40 7.80 7.60 7.70 7.60 7 90 t 0 6 2 1 0 6 3 .D PABX_BVA.M Wed O c t 10 1 3 : 0 0 : 0 6 2 0 0 0 Page 2 000802 Primedica-Worcester Project number. PABX-BVA 418-018:PAGE 1-180 Page 179 Peak Area Report Sample Name: 2 x STD-5 Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/18/200010:20 Operator: MAN Data File Name: 0621064.D Data FHe Patti: D:\MSD_DATA\PABVPASX-BVA\1-06M\ Instrument Method Name: PABX-BVA Instrument Name: MS7 Instrument Vial Number: 64 Compound 1) 2) Compound Nam MVL MVL-04 Retention Time IMIn.i 7.13 7.16 jog 131 135 P aakA raa M* 2410 4094 Pak Ama Batto 58.7E-3 M* * Th* I tu r "m" in thia column indicata* that manual Intoerabon m i pttfwrwwl on that compound. Pnnt Date and Time: 10/18/200013:00 Page 1 000803 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-181 Page 180 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621064.D Vial: 64 18 Oct 2000 10:20 2 x STD-5 Water Operator: MAN Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) |4bundane TIC: 0621064 D 2000! 1500 10005001 [\ ! I jI ! V ' ' 1 i r~T ' .1---- ' i ------I-----------r p, p -- , i . , , i . I-T| . . . . i i 1 .......... . ;---r-- p -- 6.10 6.20 _ 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7 30 7 0 7 50 7.60 770 7 80 MVL ... ... ........ Amount: 0.00________________ f Ion:'131 (7.135) ....... ................ . i Reap: 2410 790 !A -- T-- T- ----- 1........................ , , - ,-r , I'. ----- - r ----------------I--- --- 1 ' ' ' ' fT . . . I . 6.20 6 30 6.40 6.50 6.60 6.70 6.60 6.90 7.00 7.10 7.20 7.30 7.40 7.50 4 -d4 Ion: 135 (7.158) Amount : 0.00 jResp: 4094 ---- I |.,P - T ,I 7.60 7.70 7.B0 7.90 I I I 5.20 6.30 r . . , - - r r . - T - p t t r - f ----------r ' - T - T - l ----- ' i 6.40 6.50 6.60 6.70 6 80 6.90 7.00 !\ IV I 7,10 7.20 7,30 vI 7.40 r. 7.50 I . ; I I 1 ! ........... 'i - r 7,60 7.70 7.00 7.90 0 6 2 1 0 6 4 . D PABX BVA .M Wed O ct 10 1 3 : 0 0 : 2 6 2000 Page 2 000804 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-182 Page 181 Peak Area Report Sample Name: 2 x STD-0 Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/102000 10:40 Operator: MAN Data File Name: 0621065.D Data File Path: D:\MSD_DATA\PAB\PABX-BVAVI-0e2-n Instrument Method Name: PABX_BVA inatrument Name: MS7 Instrument Vial N um ber 65 Compound * 1) 2) Compound Name MVL MVL-d4 Retention Time (Min,) 7 13 7.16 ten 131 135 Peek Area M* 5207 5437 Peak Area Ratio 957.7E-3 M" a The letter "m" in thre column indicates that manual inteershon was performed on that compound. Print Date and Time: 10/18/2000 13:00 Page 1 0 0 0 8 BS Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-183 Page 182 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621065.D Vial: 65 18 Oct 2000 10:40 Operator: MAN 2 x STD-6 Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) bundartca i i 3000) !j 200! TIC: 0621065.0 A l\ !\ |\ 100Q j! I; ' W H .( I , f T *' I | I I' r f 1 r - [ I I i t ; n I r 7T" (Tww-> 6.10 6.20 6.30 5 40 6.50 660 6.70 6 80 M V L _________ :___________________ !.Ion: "131 (7.134) iReep: 5207 -i-JUrrw- ,- p , FT| . , , . | ' ^ 6.90 7.00 7 10 7.20 Amount: ................: ......................-- ; > 7.30 7,40 7,50 7,60 7.70 7.60 7.90 0.00----- ----------------------------------------------------------------------------------- J ' ' ' I ' *1 ' -- 6.20 6 30 <Q 6.50 6 60 -d4 m , Ion: 135 (7.161) |Resp: 5437 1 ' I-- " 1 ! ' 1 1 'T T 6.70 6.80 650 7.00 7.10 7 .a 730 Amount : A 7,40 7,50 7.60 7.70 7.80 7 90 0.00 _________________ i\ -n T " -T -, . Iri i "'"i 620 6.30 6.4Q 6 SO 6.60 6.70 6.60 .6,90 7.00 7 10 7 20 7.30 7 40 7.50 7.60 7.70 7 80 7,90 0 6 2 1 0 6 5 .D PABX BVA.M Wed O c t 18 1 3 : 0 0 : 4 5 2 0 0 0 Page 2 000806 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-184 Page 183 Peak Area Report Sample Name: 2 1 STD-7 Wafer Notebook Reference: PABX-8VA-1-062 Date Acquired: 10/18/2000 10:59 Operator: MAN Data File Name: 0621066.D Data File Patti: D \MSD_0ATA\PAB\PAaX-BVAU-062-1\ instrument Method Name: PABX_BVA Instrument Name: MS7 Instrument Vial Number: 66 Comoound # ii 2) Comoound Nemo MVL MVL-dd Retention Tim (Min.} 7.14 7.16 Ion 131 135 Peak Area M* 11012 6581 Peak Area Ratio 167.8E-2 M* * The letter "m" m this column indicates m et manual mtepration was performed on that compound. Print Date and Time: 10/18/200013.00 Page 1 000807 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-185 Page 184 Quantitation Report tQT Reviewed lo tt in o Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BVA\l-062-l\0621066.D Vial: 66 18 0ct~2000 10:59 Operator: MAN 2 x STD-7 Water Inst : MS7 MS Integration Pararne : RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator! lAbundanc TIC: 0621066.D -wooj j 2000! I iI Y .... . i' 6.10 6.20 6.30 6.40 MVL________ !' Ion: 131 17.135) ; Resp: 11012 !\ 6 80 6.90... 7.00 7 10 7 20 7 30 7.40 7 50 7.60 7,70 7.80 7.90 Amount : 0.00 i\ l\ _i 6.20 6.30 6.40 6.50 6.60 6.70 6.80 630 7 00 7,10__ 720 7.33 7.40 7.50 7.60 7.70 7,80 7.90 l7L-d4 _____ Amount : 0.00 Ion: 135 (7.159) Resp: 6561 / V. 6.20 630 6.40 6.50 6.60 6.70 5.80 6 5 0 7.00 7.10 7.20 730 7.40 7.50 7 60 7 70 7.80 7,90 0 6 2 1 0 6 6 .D PABX_BVA.M Wed O c t 18 1 3 : 0 1 : 0 4 2 0 0 0 Page 2 000308 418-018:PAGE 1-186 FINAL REPORT GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR ARGUS STUDY 418-018 Primedica Project Number: PABX-BBA PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 000809 418-018:PAGE 1-187 G P rimedica FINAL REPORT GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR ARGUS STUDY 418-018 Primedica Project Number: PABX-BBA Submitted to: PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 Submitted by: Primedica-Worcester 57 Union Street Worcester, MA 01608 Primedica Report # PABX-BBA-01-145 Page 1 of 187 Issue Date: November 7, 2001 Project Scientist Bioanalytical Chemistry Department Primedi ca-W orcester 00810 OPrtm edo Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-188 Page 2 Primedica Argus Study 418-018 COMPLIANCE STATEMENT This project was conducted in compliance with the Good Laboratory Practice Regulations, 21CFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision/ recommendation on compliance with principles of good laboratory practice. Official Journal of European Communities: Legislation 32 (No. L 315; 28 October): 1-17. There were no deviations from the aforementioned standards, which affected the quality or integrity of the project or the interpretation of the results in this report. Mark A. Netsch, B.A. / Date Project Scientist O O O S11 GPRTM F.nO Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-189 Page 3 Primedica Argus Study 418-018 QUALITY ASSURANCE STATEMENT The following are the inspection dates and report dates of QAU audit/inspections for GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA PLASMA FOR ARGUS STUDY 418-018, Project Number PABX-BBA. Critical Phases 1.Laboratory Procedure 2. Raw Data 3. Draft Final Report Date Inspected 01/23/2000 03/29/2001 04/19/2001 Date Report Subm itted to Study Director Management 04/27/2001 01/29/2001 04/27/2001 04/03/2001 04/27/2001 04/26/2001 The Final Report GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA PLASMA FOR ARGUS STUDY 418-018, report number PABX-BBA-01-145, was reviewed for compliance with the Good Laboratory Practice Regulations, 21CFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision/ recommendation on compliance with principles of good laboratory practice. Official Journal of European Communities: Legislation 32 (No. L 315; 28 October): 1-17, on 11/07/01. The results as presented accurately reflect the raw data. Quality Assurance ui Date 000812 GPRTMFDTf.A Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-190 Page 4 Primedica Argus Study 418-018 TABLE OF CONTENTS Page No. COMPLIANCE STATEMENT................................................................... 2 QUALITY ASSURANCE STATEMENT...........................................................................3 LIST OF TABLES AND APPENDICES.............................................................................5 CONTRIBUTING PERSONNEL.................................................................. 6 ANALYTICAL STANDARD CHARACTERIZATION/STABILITY..............................7 ARCHIVAL STORAGE...................................................................................................... 8 SUMMARY...........................................................................................................................9 Sample Receipt......................................................................................................... 9 Method Summary...................................................................................................... 9 Calibration Standard Accuracy and Precision......................................................... 9 Quality Control Sample Accuracy and Precision.................................................. 10 Study Sample Concentrations................................................................................. 10 CONCLUSIONS..................................................................................................................10 060813 GPRI/WnTfA Primedica-Worcester Project Number: PA B X -B B A Final Report 418-018:PAGE 1-191 Page 5 Primedica Argus Study 418*018 LIST OF TABLES AND APPENDICES Page No. TABLE 1 Summary of Back-Calculated Mevalonic Acid Lactone Calibration Standard Concentrations......................................................................................... 12 TABLE 2 Summary of Interpolated Mevalonic Acid Lactone QC Standard Concentrations.................................................................................... 13 TABLE 3 Summary of Mevalonic Acid Lactone Least-Squares Linear Regression Constants and Analysis Dates................................................................................. 14 TABLE 4 Summary of Mevalonic Acid Lactone Concentrations in EDTA Rat Plasma Study Samples............................................................................................ 15 APPENDIX A Representative Chromatographic Data - Batch No. PABX-BBA-1-047-1............23 000814 O PrIMFOICA Primedica-Worcester Project Number: PAJBX-BBA Final Report 418-018:PAGE 1-192 Page 6 Primedica Argus Study 418-018 CONTRIBUTING PERSONNEL Project Scientist................................................................ Director, Analytical Chemistry......................................... Analytical Chemist............................................................ Report Coordinator........................................................... ....................... Brenda L. Brooks 000815 G P rtaafdfa Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-193 Page 7 Primedica Argus Study 418-018 ANALYTICAL STANDARD CHARACTERIZATION/STABILITY Analytical Standard: Common Name: Physical Description: Lot Number: Storage Conditions: Expiration Date: Date Received: Amount Received: Supplier: Internal Standard: Common Name: Physical Description: Lot Number: Storage Conditions: Expiration Date: Date Received: Amount Received: Supplier: DL-Mevalonic acid lactone Mevalonic acid lactone or Mevalonolacetone or MVL White crystals 47300/1 50498 5 3 C, desiccate, store under nitrogen September 13, 2002 (Assigned by Primedica) September 13, 2000 5 grams Aldrich Chemical Co. DL-Mevalonolacetone-4,4,5,5-d4 MVL-d4 Clear liquid U-295 22 5C September 11, 2002 (Assigned by Primedica) September 11,2000 0.1 gram Cambridge Isotope Laboratories Characterization and Stability: The characterization of the analytical standard and internal standard is the responsibility of the Supplier, as is the method of synthesis, fabrication or derivation and stability determination. 000*16 OPRIM FOTfA Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-194 Page 8 Primedica Argus Study 418-018 ARCHIVAL STORAGE Archival Storage: The original Final Report and raw data will be maintained for a minimum period of five years following submission of the final report in the Primedica-Worcester Archives in Worcester, MA. After five years, the Sponsor will be contacted for disposition instructions. Archival materials will be indexed by Primedica-Worcester Report # PABX-BBA-01-145. 000817 G PR T M F.D F.A Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-195 Page 9 Primedica Argus Study 418-018 SUMMARY: Sample Receipt: Samples in this project were received in good condition from Primedica Argus Laboratories in four shipments between the dates of December 5, 2000 and February 13, 2001. The samples were stored at <-20C until the time of analysis. Method Summary: Samples in this project were analyzed according to the method described in Primedica Validation Report # PABX-BVA-01-62. The method involves the conversion of mevalonic acid (MVA) to mevalonic acid lactone (MVL) and the analysis of a liquid/liquid extract by gas chromatography coupled with mass spectrometry (GC/MS) using positive chemical ionization (PCI). Calibration and quality control standards were prepared in water because mevalonic acid is a naturally occurring compound in plasma and concentrations vary with diet. The internal standard, MVL-d4, was used for quantification. The method was validated for the extraction of mevalonic acid lactone from 100 pL EDTA rat plasma samples over a concentration range of 10.0 ng/mL to 250 ng/mL. The complete description of the assay and summaries of its performance during the validation can be found in the report cited above. Sample analyses for this project were initiated on January 2, 2001 and completed on February 15, 2001. See Table 3 for the analysis dates of each batch. A copy of representative raw chromatographic data from the first batch of sample analyses is provided in Appendix A. Calibration Standard Accuracy and Precision: Duplicate and bracketing seven-point calibration curves were prepared and analyzed with each batch of samples in an analytical run. For each analytical run, a l/X2-weighted least-squares linear regression was performed on the combined calibrant data set. Calculated correlation coefficients (r) for the regressions performed were >0.996 (see Table 3). Summaries of back-calculated calibration standard concentrations are provided in Table 1. Mean accuracy, expressed as %bias, ranged between 97.9% and 102.7% across concentrations and analytical runs. In addition, precision, as measured by % Relative Standard Deviation (%RSD), ranged from 2.2% to 4.9% across concentrations and analytical runs. I 000*1* O P rtmrdtca Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-196 Page 10 Primedica Argus Study 418-018 Quality Control Sample Accuracy and Precision: Summaries of interpolated Quality Control sample concentrations are provided in Table 2. Accuracy, as measured by %bias, ranged between 93.6% and 95.6%. Precision, as measured by %RSD, ranged from 2.9% to 8.9% across all concentrations. Study Sample Concentrations: Samples where no peak was detected or the interpolated concentration was less than 10.0 ng/mL before adjustments for dilution factors, the assay lower limit of quantification, were labeled as "BQL" (below quantitation limit). Some samples reported as BQL were analyzed with a dilution but due to limited sample volume could not be reanalyzed. The detection limit for these samples is the lower limit of quantification (10.0 ng/mL) multiplied by the dilution factor. One samples where the interpolated concentration was greater than 250 ng/mL before adjustment for the dilution factor, the assay upper limit of quantification, was labeled as "AQL" (above quantitation limit). This sample could not be reanalyzed due to limited sample volume. The result for this sample was estimated to be greater than 12,500 ng/mL (250 ng/mL multiplied by the dilution factor of 50). Interpolated concentrations for all remaining samples are expressed in ng/mL. Refer to Table 4 for concentrations of mevalonic acid lactone in rat plasma study samples. CONCLUSIONS: All analytical results were within acceptable limits with the exception of one value reported as an AQL estimate due to insufficient sample available for reanalysis. ooflais Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-197 Page 11 TABLES 00082 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-198 Page 12 TABLE 1 SUMMARY OF BACK-CALCULATED MEVALONIC ACID LACTONE CALIBRATION STANDARD CONCENTRATIONS Batch ID PABX-BBA-1-047-1 PABX-BBA-1-054-1 PABX-BBA-1-056-1 PABX-BBA-1-058-1 PABX-BBA-1-062-1 PABX-BBA-1-065-1 PABX-BBA-1-067-1 PABX-BBA-1-069-1 PABX-BBA-1-072-1 PABX-BBA-1-075-1 10.0 NR 10.1 10.4 9.82 10.3 9.68 9.85 9.68 9.73 NR 10.4 10.1 9.37 9.94 9.14 10.7 10.0 NR NR 9.97 Theoretical Nominal Concentrations (ng/mL) 20.0 30.0 50.0 90.0 150 19.9 29.6 48.6 94.9 20.0 NR 48.9 91.6 152 150 19.8 30.3 48.4 88.4 19.3 30.1 51.0 91.5 150 155 19.8 30.9 49.0 87.8 20.0 30.2 51.2 89.4 150 150 21.3 30.7 48.1 89.4 20.6 30.9 48.1 87.0 152 152 20.2 30.5 48.6 89.2 20.5 31.0 50.0 91.2 153 151 19.6 30.5 46.7 85.6 18.6 30.9 47.6 93.8 149 163 19.4 30.2 46.8 87.0 22.9 31.4 53.1 92.0 148 152 19.0 29.6 52.2 90.8 20.6 32.4 49.6 89.6 155 147 19.7 30.4 48.5 92.1 NR NR 51.3 92.0 144 159 NR 34.1 46.1 91.0 145 19.1 31.0 45.5 88.6 163 250 249 245 246 251 248 254 249 248 242 244 255 255 235 245 239 240 239 246 249 251 Mean Std. Dev. n %RSD %Nominal 9.95 0.396 16 4.0% 99.5% 20.0 0.972 18 4.9% 100.1% 30.8 1.05 18 3.4% 102.7% 49.0 2.02 20 4.1% 97.9% 90.1 2.37 20 2.6% 100.2% 152 5.08 20 3.3% 101.3% 247 5.54 20 2.2% 98.6% NR = Calibration standard excluded from regression, data not reported 000*21 Prixnedica-Worcester Project Number: PABX-BBA 418-018 :PAGE I-199 P a g e 13 TABLE 2 SUMMARY OF INTERPOLATED MEVALONIC ACID LACTONE QC STANDARD CONCENTRATIONS Batch ID Theoretical Nominal Concentrations (ng/mL) 25.0 70.0 200 PABX-BBA-1-047-1 22.1 53.6* 74.3 67.2 199 184 PABX-BBA-1-054-1 23.5 24.9 65.4 70.0 183 187 PABX-BBA-1-056-1 22.5 22.9 65.2 65.9 184 182 PABX-BBA-1-058-1 24.2 23.3 68.1 68.2 189 191 PABX-BBA-1-062-1 23.0 24.1 64.2 65.5 187 188 PABX-BBA-1-065-1 21.9 26.5 64.1 64.0 184 192 PABX-BBA-1-067-1 22.4 28.6 66.8 68.4 183 182 PABX-BBA-1-069-1 23.5 23.2 66.4 63.1 187 188 PABX-BBA-1-072-1 21.1 69.1 200 23.2 69.8 192 PABX-BBA-1-075-1 29.3 23.9 61.6 62.5 184 179 Mean Std. Dev. n %RSD %Nominal 23.9 2.13 19 8.9% 95.6% 66.5 3.02 20 4.5% 95.0% 187 5.46 20 2.9% 93.6% * = Failed to meet the acceptance criteria and determined to be a statistical outlier using Grubbs' Test. Value excluded from summary statistics 000422 Primedica-Worcester Project Number: PABX-BBA 418-018.-PAGE 1-200 Page 14 TABLE 3 SUMMARY OF MEVALONIC ACID LACTONE LEAST-SQUARES LINEAR REGRESSION CONSTANTS AND ANALYSIS DATES Batch ID Slone PABX-BBA-1-047-1 0.0067118 PABX-BBA-1-054-1 0.0062552 PABX-BBA-1-056-1 0.0061528 PABX-BBA-1-058-1 0.0060042 PABX-BBA-1-062-1 0.0061041 PABX-BBA-1-065-1 0.0064579 PABX-BBA-1-067-1 0.0064673 PABX-BBA-1-069-1 0.0063214 PABX-BBA-1-072-1 0.0064268 PABX-BBA-1-075-1 0.0069212 Interceot Correlation Coefficient fri Analysis Date 0.014046 0.99956 01/02/01 -0.007087 0.99953 01/17/01 -0.007974 0.99965 01/19/01 -0.006094 0.99909 01/23/01 -0.006886 0.99954 01/26/01 -0.008650 0.99802 02/01/01 -0.002970 0.99698 02/07/01 0.00018303 0.99789 02/07/01 0.0090496 0.99919 02/13/01 -0.01975 0.99652 02/15/01 000*23 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-201 Page 15 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal # -Groun #-Generation Concentration ('ns/ml.'l Batch ID 10902-GROUP 1-F0 10903-GROUP 1-F0 10904-GROUP 1-F0 10905-GROUP 1-F0 10906-GROUP1-F0 10907-GROUP 1-F0 10908-GROUP 1-F0 10909-GROUP1-F0 10910-GROUP1-F0 10911-GROUP 1-F0 10912-GROUP 1-F0 10913-GROUP 1-F0 10914-GROUP 1-F0 17.3 PABX-BBA-1-054-1 23.7 PABX-BBA-1-054-1 26.3 PABX-BBA-1-054-1 26.0 PABX-BBA-1-054-1 36.0 PABX-BBA-1-054-1 205 PABX-BBA-1-054-1 36.5 PABX-BBA-1-054-1 32.1 PABX-BBA-1-054-1 20.7 PABX-BBA-1-054-1 35.1 PABX-BBA-1-054-1 19.8 PABX-BBA-1-054-1 34.2 PABX-BBA-1-054-1 23.3 PABX-BBA-1-054-1 10901-GROUP 1-FI 10902-GROUP 1-FI 10903-GROUP1-FI 10904-GROUP1-FI 10905-GROUP 1-FI 10906-GROUP 1-FI 10907-GROUP1-FI 10908-GROUP 1-FI 10909-GROUP 1-FI 10910-GROUP 1-F 1 10911-GROUP 1-FI 10913-GROUP1-FI 37.9 36.8 BQL (DF = 10, <100 ng/mL) 40.8 44.3 BQL (DF = 10, <100 ng/mL) 146 32.7 25.2 42.8 16.1 31.6 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 PABX-BBA-1-054-1 10915-GROUP2-F0 10916-GROUP2-FO 10917-GROUP2-FO 10918-GROUP2-F0 10919-GROUP2-FO 10920-GROUP2-F0 10921-GROUP2-F0 10922-GROUP2-F0 10923-GROUP2-F0 10924-GROUP2-F0 10925-GROUP2-F0 10927-GROUP2-F0 10928-GROUP2-F0 2300 PABX-BBA-1-058-1 1550 PABX-BBA-1-058-1 1130 PABX-BBA-1-058-1 1500 PABX-BBA-1-058-1 1850 PABX-BBA-1-058-1 1140 PABX-BBA-1-058-1 99.7 PABX-BBA-1-054-1 522 PABX-BBA-1-058-1 159 PABX-BBA-1-054-1 432 PABX-BBA-1-058-1 1780 PABX-BBA-1-058-1 218 PABX-BBA-1-054-1 2060 PABX-BBA-1-058-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 000S24 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-202 Page 16 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES A n im a l # -Group #-Generation Concentration Ine/mLl Batch ID 10915-GROUP2-F1 10916-GROUP2-F1 10917-GROUP2-F1 10918-GROUP2-F1 10919-GROUP2-F1 10920-GROUP2-F1 10921-GROUP2-F1 10922-GROUP2-F1 10924-GROUP2-F1 10925-GROUP2-F1 10927-GROUP2-F1 10928-GROUP2-F1 29300 AQL (estimated 23,200) 15100 17000 24000 12900 671 168 326 17400 821 17100 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-062-1 PABX-BBA-1-062-1 PABX-BBA-1-058-1 PABX-BBA-1-062-1 PABX-BBA-1-058-1 PABX-BBA-1-054-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-062-1 10929-GROUP3-F0 10930-GROUP3-F0 10931-GROUP3-F0 10932-GROUP3-F0 10933-GROUP3-F0 10934-GROUP3-F0 10935-GROUP3-F0 10937-GROUP3-F0 10938-GROUP3-F0 10940-GROUP3-F0 10941-GROUP3-FO 10942-GROUP3-F0 158 PABX-BBA-1-047-1 1670 PABX-BBA-1-058-1 1440 PABX-BBA-1-058-1 1220 PABX-BBA-1-058-1 1440 PABX-BBA-1-058-1 8 6 6 PABX-BBA-1-058-1 1530 PABX-BBA-1-058-1 1410 PABX-BBA-1-058-1 451 PABX-BBA-1-058-1 1380 PABX-BBA-1-062-1 416 PABX-BBA-1-058-1 327 PABX-BBA-1-058-1 10929-GROUP3-F1 10930-GROUP3-F1 10931-GROUP3-F1 10932-GROUP3-F1 10933-GROUP3-F1 10934-GROUP3-F1 10935-GROUP3-F1 10937-GROUP3-F1 10940-GROUP3-F1 10942-GROUP3-F1 156 25500 28500 9770 19300 9430 13900 13400 12700 327 PABX-BBA-1-047-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor * = Estimated value, DF = > 12,500 ng/mL 000*25 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-203 Page 17 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal # -Group ^-Generation Concentration fne/mLl Batch ID 10943-GROUP4-F0 10944-GROUP4-F0 10945-GROUP4-F0 10946-GROUP4-F0 10947-GROUP4-F0 10948-GROUP4-F0 10949-GROUP4-F0 10950-GROUP4-F0 10951-GROUP4-FO 10952-GROUP4-F0 10953-GROUP4-F0 10954-GROUP4-F0 10955-GROUP4-F0 10956-GROUP4-F0 133 172 1590 2460 1600 161 760 1730 1560 99.1 1160 1000 203 24000 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-047-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-047-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-047-1 PABX-BBA-1-058-1 10945-GROUP4-F1 10946-GROUP4-F1 10947-GROUP4-F1 10949-GROUP4-F1 10950-GROUP4-F1 10951-GROUP4-F1 10953-GROUP4-F1 10954-GROUP4-F1 25300 22900 16300 20500 19000 20300 18000 19100 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 11029-GROUP11-F0 11031-GROUP 11-F0 11032-GROUP 11-F0 11033-GROUP 11-F0 11034-GROUP11-F0 11035-GROUP 11-F0 1103 6-GROUP 11-F0 11037-GROUP 11-F0 1103 8-GROUP 11-F0 61.4 PABX-BBA-1-047-1 59.7 PABX-BBA-1-047-1 22.0 PABX-BBA-1-047-1 27.5 PABX-BBA-1-047-1 797 PABX-BBA-1-058-1 37.3 PABX-BBA-1-047-1 48.0 PABX-BBA-1-047-1 38.0 PABX-BBA-1-047-1 42.6 PABX-BBA-1-047-1 11029-GROUP 11-FI 11031-GROUP 11-FI 11033-GROUP11-F1 11036-GROUP 11-FI 11037-GROUP 11-FI 17.4 PABX-BBA-1-047-1 11.4 PABX-BBA-1-047-1 20.2 PABX-BBA-1-047-1 17.8 PABX-BBA-1-047-1 15.7 PABX-BBA-1-047-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 000826 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-204 Page 18 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal # -GrouD #-Generation Concentration (ne/mLl Batch ID 11040-GROUP12-FO 11041-GROUP12-FO 11042-GROUP 12-FO 11044-GROUP 12-FO 11045-GROUP 12-FO 11046-GROUP 12-FO 11047-GROUP12-FO 11048-GROUP12-F0 11049-GROUP 12-FO 11050-GROUP12-FO 11051-GROUP12-FO 11052-GROUP12-FO 15.5 24.8 22.3 31.5 15.5 18.9 22.5 BQL (DF = 1, <10 ng/mL) 14.0 14.9 24.2 21.9 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 11040-GROUP12-F1 11044-GROUP 12-F1 11045-GROUP 12-F1 11046-GROUP 12-F1 11047-GROUP12-F1 11050-GROUP12-F1 11051-GROUP 12-F1 11052-GROUP12-F1 52.5 PABX-BBA-1-056-1 54.4 PABX-BBA-1-056-1 31.6 PABX-BBA-1-056-1 190 PABX-BBA-1-056-1 87.3 PABX-BBA-1-056-1 33.7 PABX-BBA-1-056-1 55.6 PABX-BBA-1-056-1 31.0 PABX-BBA-1-056-1 11053-GROUP 13-FO 11054-GROUP 13-FO 11055-GROUP 13-F0 11056-GROUP 13-FO 11057-GROUP 13-FO 11058-GROUP13-F0 11059-GROUP 13-FO 11060-GROUP 13-FO 11061-GROUP 13-FO 11062-GROUP13-FO 11063-GROUP 13-FO 11064-GROUP 13-FO 11065-GROUP 13-FO 11066-GROUP 13-FO 28.3 524 BQL (DF = 1, <10 ng/mL) 28.8 20.8 29.4 36.7 15.0 19.2 23.8 22.9 18.0 66.9 17.3 PABX-BBA-1-056-1 PABX-BBA-1-062-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 11053-GROUP13-F1 11054-GROUP13-F1 11056-GROUP13-F1 11058-GROUP13-F1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 32.0 199 41.3 64.0 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 000827 Primedica-Worcester Project Number: PABX-BBA 418-018 .`PAGE 1-205 Page 19 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal # -GrouD #-Generation Concentration ine/mLi Batch ED 11059-GROUP13-F1 11060-GROUP 13-F1 11062-GROUP 13-F1 11063-GROUP13-F1 11066-GROUP 13-F1 BQL (DF = 2, <20 ng/mL) 185 71.1 194 51.9 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 11501-GROUP1-FO 11502-GROUP1-F0 11503-GROUP l-FO 11504-GROUP1-F0 11505-GROUP 1-F0 11506-GROUP1-FO 11507-GROUP1-FO 11508-GROUP1-FO 11509-GROUP1-FO 11510-GROUP 1-FO 11511-GROUP1-F0 11512-GROUP1-F0 11513-GROUP1-FO 11514-GROUP1-F0 27.1 30.2 26.9 BQL (DF = 2, <20 ng/mL) 107 2 2 .8 17.8 25.5 19.0 18.9 28.0 27.2 2 0 .8 26.4 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 11515-GROUP2-FO 11516-GROUP2-FO 11517-GROUP2-FO 11518-GROUP2-FO 11519-GROUP2-FO 11520-GROUP2-F0 11521-GROUP2-FO 11523-GROUP2-FO 11524-GROUP2-FO 11525-GROUP2-F0 11526-GROUP2-FO 11527-GROUP2-FO 11528-GROUP2-F0 173 PABX-BBA-1-065-1 161 PABX-BBA-1-065-1 92.8 PABX-BBA-1-065-1 160 PABX-BBA-1-065-1 135 PABX-BBA-1-065-1 413 PABX-BBA-1-072-1 135 PABX-BBA-1-065-1 190 PABX-BBA-1-065-1 32.7 PABX-BBA-1-065-1 146 PABX-BBA-1-065-1 89.6 PABX-BBA-1-065-1 356 PABX-BBA-1-072-1 111 PABX-BBA-1-065-1 11529-GROUP3-FO 11530-GROUP3-F0 11532-GROUP3-F0 11533-GROUP3-F0 126 PABX-BBA-1-067-1 429 PABX-BBA-1-072-1 1180 PABX-BBA-1-072-1 821 PABX-BBA-1-072-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 000828 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-206 Page 20 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal # -Grouo #-Generation Concentration (ne/mL) Batch ID 11534-GROUP3-FO 11535-GROUP3-F0 11536-GROUP3-FO 11537-GROUP3-FO 11538-GROUP3-FO 11539-GROUP3-FO 11540-GROUP3-F0 11541-GROUP3-FO 11542-GROUP3-FO 11543-GROUP3-F0 128 PABX-BBA-1-067-1 8210 PABX-BBA-1-072-1 375 PABX-BBA-1-072-1 141 PABX-BBA-1-067-1 411 PABX-BBA-1-072-1 2530 PABX-BBA-1-072-1 173 PABX-BBA-1-067-1 3680 PABX-BBA-1-072-1 311 PABX-BBA-1-072-1 347 PABX-BBA-1-072-1 11544-GROUP4-FO 11545-GROUP4-FO 11546-GROUP4-FO 11547-GROUP4-FO 11549-GROUP4-FO 11550-GROUP4-F0 11551-GROUP4-FO 11552-GROUP4-FO 11553-GROUP4-FO 11554-GROUP4-FO 11555-GROUP4-FO 11556-GROUP4-FO II557-GROUP4-FO 6970 1550 9720 164 82.1 494 483 457 308000 ** 389 648 320 541 PABX-BBA-1-072-1 PABX-BBA-1-072-1 PABX-BBA-1-075-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-072-1 PABX-BBA-1-072-1 PABX-BBA-1-072-1 PABX-BBA-1-075-1 PABX-BBA-1-072-1 PABX-BBA-1-072-1 PABX-BBA-1-072-1 PABX-BBA-1-072-1 11630-GROUP11-F0 11632-GROUP11-FO 11633-GROUP11-F0 11634-GROUP11-F0 11635-GROUP11-F0 11636-GROUP 11-F0 11637-GROUP 11-FO 11638-GROUP11-F0 11639-GROUP11-FO 48.1 PABX-BBA-1-067-1 25.4 PABX-BBA-1-067-1 25.8 PABX-BBA-1-067-1 29.6 PABX-BBA-1-067-1 80.8 PABX-BBA-1-067-1 40.3 PABX-BBA-1-067-1 38.7 PABX-BBA-1-067-1 35.8 PABX-BBA-1-067-1 38.3 PABX-BBA-1-067-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor ** = Value was confirmed 000829 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-207 Page 21 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES A nim al # -Grouo #-Generation Concentration fne/ml.l Batch ID 11640-GROUP 12-FO 11641-GROUP12-FO 11642-GROUP 12-FO 11643-GROUP12-FO 11644-GROUP 12-FO 11645-GROUP 12-FO 11646-GROUP12-FO 11647-GROUP12-FO 11648-GROUP12-FO 11649-GROUP12-FO 11650-GROUP 12-FO 11651-GROUP12-FO 11653-GROUP12-FO 41.3 PABX-BBA-1-067-1 34.2 PABX-BBA-1-067-1 47.6 PABX-BBA-1-067-1 14.6 PABX-BBA-1-067-1 25.8 PABX-BBA-1-067-1 17.5 PABX-BBA-1-067-1 26.5 PABX-BBA-1-067-1 35.5 PABX-BBA-1-067-1 102 PABX-BBA-1-067-1 19.7 PABX-BBA-1-067-1 27.3 PABX-BBA-1-072-1 24.6 PABX-BBA-1-067-1 38.1 PABX-BBA-1-067-1 11654-GROUP 13-F0 11655-GROUP13-F0 11656-GROUP 13-F0 11657-GROUP 13-F0 11658-GROUP13-F0 11659-GROUP 13-F0 11660-GROUP13-F0 11661-GROUP13-F0 11662-GROUP13-FO 11663-GROUP13-FO 11664-GROUP 13-FO 11665-GROUP 13-FO 11667-GROUP13-FO 32.4 30.5 39.5 11.9 11.8 BQL (DF = 1, <10 ng/mL) 38.1 31.2 16.8 42.0 28.0 BQL (DF = 1, <10 ng/mL) BQL (DF= 1, <10 ng/mL) PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 11501/11506-GROUP 1-FI 11504-GROUP 1-FI 11502/11507-GROUP 1-FI 11505-GROUP 1-FI 15.4 BQL (DF = 4, <40 ng/mL) 19.2 BQL (DF = 3, <30 ng/mL) PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 11508/11503-GROUP1-F1 11509/11510-GROUP1-FI 11512/11511-GROUP1-F1 11513/11514-GROUP1-F1 26.4 PABX-BBA-1-069-1 25.8 PABX-BBA-1-069-1 23.3 PABX-BBA-1-069-1 20.5 PABX-BBA-1-069-1 11515/11524-GROUP2-F1 11517/11520-GROUP2-F1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 380 393 PABX-BBA-1 -072-1 PABX-BBA-1-072-1 000S30 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-208 Page 22 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES A n im a l # -GrouD #-Generation Concentration fne/mL) Batch ID 11519/11518-GROUP2-F1 11521/11516-GROUP2-F1 11523/11527-GROUP2-F1 11525/11528-GROUP2-F1 11526-GROUP2-F1 275 PABX-BBA-1-072-1 239 PABX-BBA-1-069-1 214 PABX-BBA-1-069-1 152 PABX-BBA-1-069-1 49.0 PABX-BBA-1-069-1 11536/11537-GROUP3-F1 11538/11534-GROUP3-F1 11540/11530-GROUP3-F1 11542-GROUP3-F1 11543/11529-GROUP3-F1 242 PABX-BBA-1-069-1 395 PABX-BBA-1-072-1 193 PABX-BBA-1-069-1 243 PABX-BBA-1-069-1 340 PABX-BBA-1-072-1 11547-GROUP4-F1 11552-GROUP4-F1 11630/11633-GROUP 11-FI 11632/11636-GROUP 11-F1 11635/11637-GROUP11-FI 11638-GROUP11-F1 11639/11634-GROUP 11-FI 51.3 339 21.7 24.0 BQL (DF = 2, <20 ng/mL) BQL (DF = 2, <20 ng/mL) 21.4 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 11640/11641 -GROUP 12-F1 11643/11647-GROUP 12-F 1 11646/11644-GROUP12-F1 11648/11651-GROUP 12-F 1 11650-GROUP 12-F 1 11653/11642-GROUP 12-F 1 23.3 BQL (DF = 4, <40 ng/mL) 14.7 21.9 22.4 BQL (DF = 2, <20 ng/mL) PABX-BBA-1-069-1 PABX-BBA-1-072-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-072-1 PABX-BBA-1-069-1 11654/11662-GROUP 13-F1 11656/11655-GROUP 13-F1 BQL (DF = 2, <20 ng/mL) BQL (DF = 2, <20 ng/mL) PABX-BBA-1-069-1 PABX-BBA-1-069-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 000S31 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-209 Page 23 APPENDIX A REPRESENTATIVE CHROMATOGRAPHIC DATA - BATCH NO. PABX-1-047-1 000832 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-210 Page 24 Peak Area Report Sample Nam: 1x Blank Notebook Retereoce: PABX-BBA-1-047 Date Acquired: 1/2/01 15:29 Operator Data Fila Nam: 0471001.D Instniment Nam: MS7 Data File Path: O:\MSD_DATA\PAB\PABX-BBA\1-047-1\ btstrument Vial Number 1 Instrument Method Nam: PABX_BBA Comoound # 1) 2) Comoound Name MVL MVL*d4 Retention Time (Min.) 0.00 0.00 ion 131 135 Peak Area M* 0 0 Peak Area Rabo #DIV/0! M* The letter " n r In brie cohjmn IndicatBe that manuel Integration was performed on that compound. Print Date and Tene: 1/18/01 14:44 Page 1 000833 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-211 Page 25 D ata F ile A cq On Sam ple M ise D :\M S D D A T A \P A B \ P A B X -B B A \ l-0 4 7 -l\0 4 7 10 0 1.D V ia l: 1 2 Jan 2001 l x Blank 15 :2 9 O p erator: Inst : MS7 M V P________________ Io n : 13 1 (0.000) Amount : Not Found 0471001.D PABX BBA.M Thu Jan IB 14:44:41 2001 Page 2 000634 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-212 Page 26 Peak Area Report Sample Name: 1 X STD 1 Notebook Reifence: PABX-BBA-1-047 Data Acquired: 1/2/01 15.48 Operator: Data File Name: 0471002.D Instrument Name: MS7 Data File Patti: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number. 2 kistrument Method Name: PABX_6BA Comoound K 2) ComDOund Name MVL MVL-CW Retention Time (Min.) 7.02 7.04 ton 131 135 Peak Area M* 155 1282 Peak Area Rabo 120.9E-3 M* The tetter *m" In this column Indicates that manual Integration teas performed on that compound. Prinl Dale and Time: 1/18/01 14:44 Page 1 0IM3S Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-213 Page 27 Data F ile Acq On Sample M ise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471Q02.D V ial: 2 2 Jan 2001 1 5 :4 8 O p erator: 1 X STD 1 Inst : MS7 MVL_______________________________________________ Amount:________ 0 .0 0 Ion : 131 17.0 19 )' 0471002.D PASX BBA.M Thu Jan 18 14:45:12 2001 Page 2 000836 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-214 Page 28 Peak Area Report Sample Name: 1 X STD 2 Notebook Reference: PABX-BBA-1-047 Date Acquired: 172/01 16:08 Operator Data FileName: 0471003.D kutniment Name: MS7 Data File Path: D:WSD_DATA\PAB\PABX-BBA\1 -047-1\ Instrument Vial Number 3 Inhument Method Name: PABX.BBA ComDOund * 1) 2) ComDound Name MVL MVL-44 Retention Time (Min.) 7.02 7.04 ion 131 135 Peak Area M* 213 1441 Peak Area Rado 147.8E-3 M* The letter "m" In this coturni Indicate that manual kitagrabon m s performed on that compound. Ptmt Date and Time: 1/18/01 14:45 Page 1 000837 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-215 Page 29 Data F ile Acq On Sample M ise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471003.D V ia l: 3 2 Jan 2001 16:08 O perator: 1 X STD 2 In st : MS7 MVL________________________________________________Amount :________ 0.00 . Io n : 131 (7.020) 0471003.D PABX BBA.M Thu Jan 18 14:45:31 2001 Page 2 000838 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-216 Page 30 Peak Area Report Sample Name: 1 X STO 3 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2AJ1 16:28 Operator Data File Name: 0471004.0 kismiment Name: MS7 Data File Path: D:'WSO_OATA\PAB1PABX-BBAM -047-1\ Instrument Vial Number. 4 Instrument Method Name: PABX_BBA Comoound # 1) 2) ComDound Nam MVL MVL-cM Retention Time (Mln.i 7.02 7.04 Ion 131 135 Peak Area M* 242 1138 Peak Area Ratio 212.7E-3 M* - The lattar "m" In thiscolum n Indie s e n th at manual Integration mbs performed on that com pound. Print Date and Time: 1/18AJ1 14:45 Page 1 Primedica-Worcester Project Number: PABX-BBA 418-018'.PAGE 1-217 Page 31 Daca F i l e Acq On Sample Mise D:\MSD DATA\ PAB\PABX-BBA\ 1 - 0 4 7 - 1 \ 0471004.D V i a l : 4 2 Jan 2001 16 :2 8 O p erator: 1 X STD 3 Inst : MS7 0471004.D PABX BBA.M Thu Jan 18 14:48:51 2001 Page 2 000846 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-218 Page 32 Peak Area Report Sample Name: 1 X STD 4 Notebook Reference: PABX-BBA-1-047 Date A cquired: 1/2/01 16:47 O p e ra to r: Data Hie Name: 0471 OOS.D Data Hie Patti: D:\MSD_DATA\PAB\PABX-BBA\1-047-11 kistmment Method Name: PABX.BBA Instniment Name: MS7 Instniment Vial Number. 5 ComDound # 1) 2) Comoound Name MVL MVL-4 Ritentlon Time fMm.l 7.02 7.04 ton 131 135 Peak Area IT 480 1412 Peak Area Ratio 339.9E-3 M` - The letter "m" in this column Indlcatee that manual Integration w as performed on that compound. Pnnt Dale and Time: 1/18/01 14:46 Page 1 000841 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-219 Page 33 Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471005.D V ia l: 5 2 Jan 2001 1 X STD 4 16:47 O perator: In st : MS7 MVIi________________________________________________Amount :________0.00 Ion: 131 (7.020) 0471005.D PABX BBA.M Thu Jan 15 14:46:11 2001 Page 2 000842 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-220 Page 34 Peak Area Report Sample Name: 1 X STD S Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 17:07 Operator. Data Rie Name: 0471006.D Instrument Name: MS7 Data Rie Patti: D:\MSD_DATA\PAB\PABX-BBAU-047-n Instrument Vial Number 6 Instrument Method Name: PABX_BBA Comoound * 1) 2) Comoound Name MVl MVL<)4 Retention Time (Mln.l 7.02 7.04 tea 131 135 Peak A m M* 790 1213 Peak Area Ratio 651.3E-3 M* a The letter "mBIn this column indicates that manual Integration wee p a r t mad on that compotmo. Print Dale and Time: 1/18/01 14:46 Page 1 000843 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-221 Page 35 Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471006.D V ial: 6 2 Jan 2001 1 7 :0 7 O p era tor: 1 X STD 5 Inst : MS7 I n t e g r a t i o n Param s: RTEINT.P a n t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) M V L _________________________________________ Amount:________ 0,00 Io n : 131 <7.017) R e s p : 790 A __JL in 'l l - 6.20 d4 6.30 6.40 on: 135 (7.044) j^ s p : 630 6.60 6.70 6.00 6.90 7.00 Amount: 7.10 7.20 0.00 7.30 7.40 7.50 7.60 7.70 7.80 7.90 i ! ' I1 1' 1 I ......... I I r. I 1 'I 111 I... T I.... I ......................! .... ..... I... II, 6180 6.30__JM0 6.50 6.60 8.70 6.80 6.90 7.00 7 10 7.20 7.30 7.40 7.50 7.60 7.70 7.60 7.90 0471006.D PABX BBA.M Thu Jan 18 14:46:30 2001 Page 2 000844 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-222 Page 36 Peak Area Report Sample Name: 1 X STD 6 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 17:27 Operator Data File Nome: 04710Q7.D ktsbumenl Name: MS7 Data Hie Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-U Inatniment Vial Number. 7 Instrument Method Name: PABX_BBA Compound 4 H 2) Compound Nam MVL WVL-04 Retention Time Min.) 7.02 7.04 km 131 135 Peak A m a M* 1257 1218 Peak'Area Ratio 103.2E-2 M* The letter "m" In IN e column Indicatee th at menue! integration erne performed on that compound. Print Dale and T u : 1V18/01 14:46 Page 1 000845 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-223 Page 37 Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471007.D V ia l: 7 2 Jan 2001 1 7 :2 7 O p erator: 1 X STD 6 In st : MS7 I n t e g r a t i o n Params : RTEINT. P an t Method : D: \HPCHEM\1 \METHODS\ PABX_BBA. M (RTE I n t e g r a t o r ) M V L _______________________________________ Amount :_____________0 .00 Ion : 131 (7.018) 0471007.D PABX BBA.M Thu Jan 18 14:46:50 2001 Page 2 000S46 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-224 Page 38 Peak Area Report Sampta Nam: 1 X STD 7 Notebook Reference: PABX-BBA-1-047 Bate Acquired: 1/201 17:47 Operator. Date Fite Name: 0471008.D Data File Path: O:\MSCLOATA\PABiPABX-8BAU-047-n Instrument Method Name: PABX.BBA Instrument Name: MS7 Instrument Vial Number 8 Comoound # 1) 2) Comoound Name MVL MVL-4 Retention Time fMin.l 7.02 7.04 Ign 131 135 P u k Area M* 2166 1264 Peek Area Ratio 168.7E-2 M* = The laner *m* kithiscolumn jndlratai thatmanual kilegialienwas partarmad on thatcompound. Print Date and Tin e: 1/18/01 14:47 Page 1 000*47 Primedica-Worcester Project Number: PABX-BBA 418-018 :PAGE 1-225 Page 39 Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1- 0 4 7 - 1 \0471008 .D V i a l : 8 2 Jan 2001 1 7 :4 7 O p era tor: 1 X STD 7 Inst : MS7 M V L _______________________________________________Amount :________0.00 Ion: 131 (7.01B) 1 Resp : 2166 y\ . . / L i f r *M ' ' ' 1 i 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7 10 7.20 730 7.40 7.50 7.60 7.70 7.80 7.90 MVL-d4 Amount : 0.00 on: 135 (7.042) 1284 .A 6-20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 720 7.30 7.40 7.50 7.60 7.70 7.80 7.90 0471008.D PABX BBA.M Thu Jan 18 14:47:10 2001 Page 2 000*48 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-226 Page 40 Peak Area Report Sample Nam: 1 X C O N TROL Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 18:07 Operator. Date Rte Nam: 0471009.0 Instrument Nam: MS7 Data Hie Path: O:\MSO_DATA\PAB\PABX-BBA11-047-U InstrumentVial Number 9 Instrument Method Name: PABX.BBA Compound 1) 2) Compound Name MVL MVL-4 Retention Time (Mln.1 0.00 7.04 Ion 131 135 Peak Area M* 0 1087 Peak Area Ratio OOO.OE-3 M* T h e letter "m" in tlita column Indicate th at menuet Intejrettun wee performed on that compound. Print Date and Time: 1/18/01 14:47 Page 1 000*49 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-227 Page 41 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471009.D Vial : 9 2 Jan 2001 18:07 Operator: 1 X CONTROL Inst : MS7 MVL_______________ Ion: 131 (0.000) Amount : Mot Found 0471009.D PABX BBA.M Thu Jan 18 14:47:30 2001 Page 2 000850 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-228 Page 42 Peak Area Report Sample Name: 1 X O C L O W N o te b o o k R e fe re n c e : PABX-BBA-1-047 Data Acquired: 1/2/01 18:26 Operator Data File Name: 0471014.D Instrument Name: MS7 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number: 14 Instrument Method Name: PABX_BBA Comoound * U 2) Comoound Name MVL MVL-CM Retention Time (Min.) 7.02 7.04 ten 131 135 Peak Area M* 184 1135 Peak Area Ratio 162.1E-3 M* The Iettar "m* In th is column indicates th at manual kitai stlo n ms performed on that compound. Print Dale and Time: 1/18/01 14:48 Page 1 oooa r / Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-229 Page 43 Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\l-047-l\0471014.D Vial: 14 x12 Jan QC 2001 LOW 18:2 Operator: Inst : MS7 0471014.D PABX BBA.M Thu Jan 18 14:49:08 2001 Page 2 000852 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-230 Page 44 Peak Area Report Sample Name: 10930-GROUP3-FO Notebook Reference: PABX-BBA-f-047 Date Acquired: 1/2A31 18:46 Operator. Data FileName: 0471025.0 Inatrument Name: MS7 Data File Peth: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ Inatrument Vial Number: 25 Inatrument Method Name: PABX.BBA Comoound t H 2) Comoound Name MVL MVL-d4 Retention Time fMin.l 7.02 7.04 Ion 131 135 Peak A m M* 6106 706 Peak Area Ratio B64.9E-2 M* T h e letter *m' In dii cokimn Indi cete th at m anual Inla y alkm we p e rt nm l on that compound. Print Oate and Time: 1/18701 14:53 Page 1 000353 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-231 Page 45 Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\l-047-l\0471025.D Vial: 25 2 Jan 2001 1B:46 Operator: 10930-GROUP3-FO Inst : MS7 M V L __________ ______ _________________ Amount :______ OJIO 'ion: 131 {7.015) !Resp: 6106 0471025.D PABX BRA.M Thu Jan 18 14:53:42 2001 Page 2 000854 Priniedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-232 Page 46 Peak Area Report Sample Name: 10931-GROUP3-FO Notebook Retarence: PABX-BBA-1-047 Date Acquired: 1/2/01 19:06 Operator Data FileName: 0471026.0 Instrument Name: MS7 Data FilePath: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number 26 Instrument Method Name: P A B X B B A Comoound # 1) 2) Comoound Name MVL MVL-d4 Retention Time (Min.) 7.01 7.04 ton 13t 135 Peek Area M* 5583 567 Peak Area Ratio 984.7E-2 M* The letter *m' In this co tu rn i Indicete that manual Integration earn performed on that compound. Preti Date and Time: 1/18/01 14:53 Page 1 000855 Priraedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-233 Page 47 Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\1-047-1\0471026.D Vial: 26 2 Jan 2001 19:06 Operator: 10 931-GROUP3 -FO Inst : MS7 k3 Integration Params: RTEINT.P ^uant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0471026.D PABX BBA.M Thu Jan IB 14:54:02 2001 Page 2 000851 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-234 Page 48 Peak Area Report Sample Name: 10932-GROUP3-FO Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 19:26 Operator: Data Ble Name: 0471027.D Instrument Name: MS7 Data Ble Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1V Instrument Vial Number: 27 Instrument Method Name: PABX_BBA Comoound # 1) 2) Comoound Narqe MVi MVL-cW Retention Time (Mln.1 7.02 7.04 Ion t31 135 Peak Area IT 5389 792 Peak Area Ratto 680.4E-2 M* The letter*m* Inthiscolumn Indicatesmat manual integrationm s performed en mat compound. Print Dale and Tane: 1/18/01 14:54 Page X 000857 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-235 Page 49 U u aU L ^L B L JlU lt VU* R U O U / Data File Acq On Sample Mise D:\MSD DATANPAB\PABX-8BA\ 1-047-110471027.D Vial: 27 2 Jan 2001 19:26 Operator: 10932-GROUP!-FO Inst ; MS7 Integration Params: RTEINT.P ant Method : D :NHPCHEMN1NMETHODSN PABX_BBA.M (RTE Integrator) 0471027.D PABX BBA.M Thu Jan IB 14:54:22 2001 Page 2 000*58 Primedica-Worcester Project Number: PABX-BBA 418-018 :PAGE 1-236 Page 50 Peak Area Report Sample Name: 10933-GROUP3-FO Notebook Reference: PABX-8BA-1-47 Dele Acquired: 1/2/01 19:45 Operator Data File Name: 0471028.0 Instrument Name: MS7 Data File Path: D:\MSD_DATA\PABV'ABX-BBAVt -047*1\ Instrument VtaiNumber 28 Instrument Method Name: PABX.BBA Comoound # 1) 2) Comoound Name MVL MVL*d4 Retention Time (Min.) 7.02 7.04 Ion 131 135 Peak Area M* 7053 515 P*ak A n a Ratio 137.0E-1 M* a The letter am a In this column Indlcelee th at m anual Integration performed on ttwt compound. Phnt Date and Time: 1/18/01 14:54 Page 1 0085S Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-237 Page 51 ftevicneu/ Data File Acq On Sample Hiac D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471028.D Vial: 2B 2 Jan 2001 19:45 Operator: 10933 -GROUP3 -FO Inst : MS7 Integration Params: RTEINT.P ant Method : D :\HPCHEM\l\METHODS\PABZ_BBA.M (RTE Integrator) 0471028.D PABX BBA.M Thu Jan 18 14:54:41 2001 Page 2 000860 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-238 Page 52 Peak Area Report Sample Name: 10934-GROUP3-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/21 20:05 Operator Data FileName: 0471029.D kiatrument Name: MS7 Data Rie Patti: D:\MSD_DATAIPAB\PA8X-BBA\1 -047-11 Inatrument Vial Number 29 Initnjment Method Name: PABX.BBA Comoound 4 i 2) Comoound Name MVL MVL-04 Retention Time fMin.l 7.01 7.04 Ion 131 135 Peak Area M* 3213 646 Peak Area Ratio 497.4E-2 M` - The letter *m* In thle column Indicete m at manual Integration w m partormed on m at compound. Print Date and Time: 1/18/01 14:54 Page 1 000SG1 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-239 Page 53 yuancrcation Report iy: jteviewea: Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471029.D Vial: 29 2 Jan 2001 20:05 Operator: 10934-GROUP3-FO Inst : MS7 Integration Pararas: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0471029.D PABX BBA.M Thu Jan 18 14:55:01 2001 Page 2 000862 Primedica-Worcester Project Number: PABX-BBA 418-018 :PAGE 1-240 Page 54 Peak Area Report Sample Name: 10935-GROUP3-FO Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 20:25 Operator: Data Ble Name: 0471030.D kwtrument Name: MS7 Data File Path: D:\MSD_DATA\PAB\PABX-BBAM-047-1\ kiatrument VialNumber 30 Instrument Method Name: PABX_BBA ComDound 1) 2) Comoound Name MVL MVL-d4 Retention Time iMIn.l 7.02 7.04 Ion 131 135 E a s M f f M* 6310 596 Peak Area Ratio 105.9E-1 M* * The letter V In Itila column indcales th a t manual Integration wee performed on that co n fo u n d . Print Dale and Time: 1/18/01 14:55 Page 1 000863 Primedica-Worcester Project Number. PABX-BBA 418-018:PAGE 1-241 Page 55 uuantitation Report iut Reviewear Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471030.D Vial: 30 2 Jan 2001 20:25 Operator: 10935-GRQUP3-FO Inst : MS7 Integration Pararne : RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL____________________ _____ ______ _____ Amount : 'ion: 131 (7.016) Reap: 6310 0.00 6.20 6.30 6-40 6.50 6.60 70 6.80 6.90 7. 7.10 720 7.30 7.40 '7.50 ` 7.60 7.70 7.80 7.90 ' '' '! M',T.-d4 Amount: 0.00 0471030.D PABX BBA.M Thu Jan 18 14:55:20 2001 Page 2 000SS4 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-242 Page 56 Peak Area Report Sample Name: 10937-GROUP3-FO Notebook Reference: PA8X-SBA-1-047 Date Acquired: 1/2/01 20:45 Operator: Data File Name: 0471031.D Instrument Name: MS7 Data File Path: D:\MSD_DATA\PA8\PABX-BBAU-047-1\ tnatrument Vial Number: 31 Instrument Method Name: PABX_BBA Compound # U 2) C o m sound Name MVL MVL-d4 Retention Time (Mln.t 7.02 7.04 2 131 135 Peak Area M* 6228 992 Peak Area Ratio 627.8E-2 M' > The letter "m" In this column inrticen that m enuel Integration was performed on that compound. Print Date and Time: 1/18/01 14:55 Page 1 000865 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-243 Page 57 yuanticacion Report lY i itev iew eu : Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471031.D Vial 31 2 Jan 2001 20:45 Operator 10937-GROUP3-FO lost MS 7 k3 Integration Params: RTEINT.P Plant Method : D:\HPCHEW\l\METHODS\PABX_BBA.M (RTE Integrator) MVL Amount : 0.00 0471031.D PABX BBA.M Thu Jan 18 14:55:40 2001 Page 2 000866 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-244 Page 58 Peak Area Report Sample Name: 10940-GROUP3-FO Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 21:04 Operator. Data FileName: 0471032.0 Instrument Name: MS7 Data Frie Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number: 32 Instrument Method Name: PABX.BBA Comoound # i) 2) Comoound Name MVL MVL-d4 Retention Time fMin.1 7.02 7.04 isa 131 135 Peak Arai M* 7133 602 Peak Area Ratio 118.5E-1 M* The letter *m* In this column indicstas H ut manual In tsy ah o n was performed on trial compound. Print Date and Time: 1/18/01 14:55 Page 1 000867 Primedica-Worcester Project Number. PABX-BBA 418-018:PA G E 1-245 Page 59 K tipU X L \\Jl K t t v i e w e u j Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471032.D Vial: 32 2 Jan 2001 21:04 Operator: 10940-GROUP3-FO Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\KETHODS\PABX_BBA.M IRTE Integrator) MVL Amount : 0.00 0471032.D PABX BBA.M Thu Jan IB 14:56:00 2001 Page 2 000868 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-246 Page 60 Peak Area Report Sample Name: 10930-GROUP3-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/D1 2124 Operator. Data File Name: 0471033.D Inetrument Name: MS7 Data File Path: D:\MSD_DATA\PAB\PABX-8BA\1-047-1\ Inetniment Vial Number 33 Instrument Method Name: PABX_BBA Comoound # U 2) Comoound Name MVL MVL-d4 Retention Time (Min.) 7.02 7.04 ton 131 135 Peak Area M* 124493 779 Peak Area Ratio 159.8E+0 M* m The letter *m* In this column kwHcats t th at manual Integration w as performed on that compound. Print Date and Time: 1/18/01 14:56 Page 1 000869 Primedica-Worcester Project Number: PABX-BBA 4 1 8 -0 1 8 .-PAGE 1-247 Page 61 Quantitation Report ((jr Reviewed) Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\1-047-1\0471033.D Vial: 33 2 Jan 2001 21:24 Operator: 10930-GROUP3-FI Inst : MS7 Integration Params: RTEINT.P Plant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) HVL________________________ ____ _______ Amount :_______ 0.00 fon: 131 (7.016) :Reap: 124493 0471033.D PABX BBA.M Thu Jan IB 14:56:20 2001 Page 2 000870 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-248 Page 62 Peak Area Report Sample Name: 10931-GROUP3-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 112/01 21:44 Operator: Data FileName: 0471034.D Instrument Name: MS7 Data Hie Path: O:\MSO_DATAVPAB1PABX-BBA\1-047-1\ Instrument Vial Number. 34 Instrument Method Name: PABX.BBA Comoound * 1) 2) Comoound Name MVL MVL-d4 Retention TimefMin.l 7.02 7.04 km 131 135 Peak Area M* 148288 815 Peak A m Ratio 181.9E+-0 M" T h e letter *m ' In th is cokim n indicslas th a t manual Integration mbs performed o n th at compound. Prinl Date and Time: 1/18/01 14:56 Page 1 000871 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-249 Page 63 Quantitation Report IQT Reviewed) Data File Acq On Sample Mise D:\MSO DATA\PAB\PABX-BBA\l-047-l\0471Q34 .D Vial: 34 2 Jan 2001 21:44 Operator: 10931-GROUPS-FI Inst : MS7 Integration Params: RTEINT.P ant Method : D :\HPCHEM\ 1\METHODS\PABX_BBA.M (RTE Integrator) TIC: 0471034.D M V L __ ________________________________ Amount :_______ 0 00 ion : 131 (7.016) 1 Resp: 148288 Amount: 0.00 0471034.D PABX BBA.M Thu Jan 18 14:56 :39 2001 Page 2 000872 Primedica-Worcester Project Number: PABX-BBA 4 1 8 -0 1 8 :PA G E 1-250 Page 64 Peak Area Report Sample Naim: 10932-GROUP3-F1 Notebook Reference: PABX-BBA-1-047 Data Acquired: 1/2/01 22:03 Operator Data FileNairn: 0471035.0 Instrument Name: MS7 Data Rie Path: D:\MSD_DATA\PAB\PABX-BBAU-047-1\ Instrument VialNumber 35 Instrument Method Name: PABX.BBA Comoound # 1) 2) Comoound Name MVL MVLJ4 Retention Time <Mln.) 7.02 7.04 las 131 135 Peak Area M* 44011 655 Peak Area Ratio 671.9E-1 M* m The letter "m" In this column Indicates th a t manual Integration m b performad on that compound. Print Date and Time: 1/18/01 14:56 Page 1 000873 Primedica-Worcester Project Number: PABX-BBA 418-018:P A G E 1-251 Page 65 uuantitation eporc ihoc Keviewecw Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471035.D Vial: 35 2 Jan 2001 22:03 Operator: 10932-GROUP3-Fl Inst : MS7 Integration Pararne : RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BHA.M (RTE Integrator) TIG. 047103533 MVL_____________________________________ Amount :______ 0 .00 Ion: 131 (7.016) Resp: 44011 0471035.D PABX BBA.M Thu Jan 18 14:56:59 2001 Page 2 000874 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-252 Page 66 Peak Area Report Sample Name: 1X Q C MIO Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 22:23 Operator: Data File Name: 0471012.D toatrament Name: MS7 Data File Patti: D:\MSD_DATA\PAB\PABX-B8A\1-047-1\ Instrument Vial Number 12 Instrument Method Name: PABX.BBA Comoound # 1) 2) Comoound Name MVL MVL-d4 Retention Time iMin.l 7.02 7.04 Ion 131 135 Peek Are M* 522 101B Peak Area Ratio 512.BE-3 M* b The letter "m" in this column Indicete that manual Integration waa parfom tad on that compound. Print Date and Time: 1/18/01 14:48 Page 1 000875 Primedica-Worcester Project Number: PABX-BBA 418-018:P A G E 1-253 Page 67 U u a u u ib a tiu ii w a b i b u Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-l\0471012.D Vial: 12 2 Jan 2001 22:23 Operator: 1 X QC MID Inst : MS7 Integration Parame: R7EINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL _ __ ______ __________ __________ Amount :______ 0.00 ! ion:" 131 (7.016) Reap: 522 A K ;.... i1 'i...... '1! '1 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 720 7.30 7.40 7.50 7.60 7.70 7.80 7.90 MVL-d4 Amount : 0.00 135 (7.043) ^ K s p : 1018 JL_ . . ,.............. 1r-i-p'F .' 1' 1 ' ''1'''1 ' '<1 '' '' 1* ''I' ''1'..... I11 1? 1 ''' 1'1 1' 11'' '1... . 1 6.20 6.30 6.40 6.50 6.60 6 70 6 80 6.90 7.00 7.10 720 7-30 7.40 7,50 7.60 7.70 7.B0 7.90 0471012.D PABX BBA.M Thu Jan 18 14:48:29 2001 Page 2 000876 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-254 Page 68 Peak Area Report Sample Name: 10933-GROUP3-F1 Notebook Reterence: PABX-BBA-1-047 Date Acquired: 1/2/01 22:43 Operator Data File Name: 047103S.D Inatniment Name: MS7 Data Hie Path: D:\WSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number 36 Instrument Method Name: PABX_BBA Comoound # 1) 2) Comoound Name MVL MVL-d4 Retention Tim (Min.) 7.02 7.04 ion 131 135 Peak Area M* 106656 m 783 Peak Area Ratio 136.2E+0 M* The letter'm' Inthis cokimn IndlcalaePlatmanual Imagrstionwaa performedon thatcompound. Prinl Date and Time: 1/18/01 14:57 Page 1 000877 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-255 Page 69 Uuancitation Report IUT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471036 .D Vial: 36 2 Jan 2001 22:43 Operator: 10933-GRODP3-Fl Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) Arasi*:;- '~ TIC:0471036.0 MVL Ion: 131 (7.016) Resp: 1066S6 MVT,-d4 Amount : __0.00 Amount: 0.00 0471036.D PABX BBA.M Thu Jan IB 14:57:19 2001 Page 2 000878 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-256 Page 70 Peak Area Report Sample Nam: 10934-GROUP3-F1 Notebook Relerenee: PABX-BBA-1-047 Date Acquired: 1/2/01 23:02 Operator Data File Name: 0471037.D Instrument Name: MS7 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number 37 Instrument Method Name: PABX_BBA Comoound t H 2) Comoound Name MVL MVL-d4 Retention Time (Min.) 7.01 7.04 ten 131 135 Peak Atea M' 46551 659 Peak Area Ratio 706.4E-1 M* The letter "m* In this column Indlcstas Biat menuet Integration aies performed on that compound. Print Dale and Tane: 1/18/01 14:57 Page 1 000*73 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-257 Page 71 Data File Acq On Sample Mise D:\MSD DATANPABNPABX-BBANl-047-1\0471037 .D Vial: 37 2 Jan 2001 23:02 Operator: 10934-GROUP3-F1 Inst : MS7 > Integration Params: RTEINT.P tant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) TTC:047137.D " MVL___________________ __________________ Amount :______ 0 .00 . Ion: 131 (7.12) MVL-d4 Amount: 0.00 ! 0471037.D PABX BBA.M Thu Jan 18 14:57:38 2001 Page 2 000880 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-258 Page 72 Peak Area Report Sample Name: 10935-GROUP3-F1 Notebook Reference: PABX-BBA-1 -047 Date Acquired: 1/2/01 23:22 Operator Data File Name: 0471038.D Inetniment Name: MS7 Data File Path: D:\MSD.DATA\PAB\PABX-6BA\1 -047-1N Instrument Vial Number 38 instrument Method Name: PABX_BBA Comoound * 1) 2) Comoound Name MVt MVL-4 Retention Time fMln.l 7.02 7.04 ion 131 135 Peak Area M* 65000 694 Peak Area Ratio 950.8E-1 SI* The letter*m* inthiscolumn Indicatesthatmanual integrationaresperformedon thatcompound. Print Date and Time: 1/18/01 14:57 Page 1 000881 Primedica-Worcester Project Number: PABX-BBA 418-018:P G E 1-259 Page 73 Data F ile A c q On Sample M ise D:\MSD D A TA\PAB\PABX-BBA\l-047-l\047103B.D V ia l : 38 2 Jan 2001 23:22 O perator: 1 0 9 3 S-GROUP3-Fl Inst : MS7 MVL A m ount: 0.00 M V L - d 4 ______________________________________A m o u n t :________ O^OO, ->n: 1 3 5 ( 7 . 0 4 0 ) 6.20 6.30 6.40 6.50 6.60 6.70 8.80 6JO 7.00 7.10 7.20 730 7 40 7.50 7.80 7.70 7J0 T M 0471030.D PABX BBA.M Thu Jan 18 14:57:58 2001 Page 2 000882 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-260 Page 74 Peak Area Report Sample Name: 10937GROUP3-F1 Notebook Reference: PA8X-BBA-1-047 Data Acquired: 1/2/01 23:42 Operator Data File Name: 0471039.D Instrument Name: MS7 Data File Patti: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1V kistrument Vial Number 39 Initmment Mettiod Name: PABX_BBA ComDOund # 1) 2) Compound Name MVL MVL-04 Retention Time /Mln.i 7.02 7.04 ion 131 135 Peak Area M* 76789 751 Peak Area Ratio 102.2E+0 M* * The M a r *m`m ilecoiiimnindicatesthatmanual Integrationm performed on thatcompound. Print Date and Time: 1/18/01 14:58 Page 1 000813 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-261 Page 75 D ata F ile A c q On Sample M ise D :\M S D D A T A \P A B \P A B X -B B A \1 -0 4 7 -1 \0 4 7 1 0 3 9 .D V i a l : 39 2 Ja n 2001 23:42 O perator: 1O937GRO0P3-F1 In s t : MS7 MVL__________ __ __________________________________ A m o u n t :________ 0 . 0 0 ' Io n : 131 '17.017) MVL-d4 Amount: 0.00 0471039.D PABX BBA.M Thu Jan IB 14:5B:1B 2001 Page 2 000884 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-262 Page 76 Peak Area Report Sample Name: 10940-GROUP3-F1 Notebook Reference: PABX-BBA-1-047 Dale Acquired: 1/3/01 0:01 Operator: Data File Name: 0471040.D Instrument Name: MS7 Data File Path: D:1MSD_DATA\PAB\PABX-SBA\1 -047-1\ Instrument Vial Number. 40 Instmmenl Method Name: PABX_BBA ComDOund * 1) 2) Comoound Nam MVl MVL-d4 Retention Time (Min.1 7.02 7.04 ton 131 135 Peak Area M* 73060 712 Peak Area Ratio 102.6E.fO M* a The lettsr *m* In this column Indicates that manual Integration was performed on that com pound. Print Date and Time: t/18/0t 14:58 Page 1 000885 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-263 Page 77 Data F ile A c q On Sample M ise D:\MSD DATA\PAB\PABX-B B A \1 - 0 4 7 - 1 \ 0 4 7 1 0 4 0 . D V i a l : 40 3 Jan 2001 00:01 O perator: 10940-GROUP3-F l In s t : MS7 I n te g r a tio n Param s: RTEINT.P a n t M ethod : D:\HPCHEM\1\METH0DS\PABX__BBA.M (RTE I n t e g r a t o r ) MVL WVL-d4 Amount: 0.00 Amount: 0.00 0471040.D PABX BBA.M Thu Jan IB 14:58:38 2001 Page 2 000881 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-264 Page 78 Peak Area Report Sample Name: 10929-GROUP3-F0 Notebook Reference: PABX-88A-1-047 Oate Acquired: 1/3/01 0:21 Operator Data Rie Name: 0471041.0 instrument Name: MS7 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ instnimenl Vial Number: 41 Instrument Method Name: PABX.BBA Compound # i) 2) Comoound Name MVL MVL-4 Retention Time (Min.) 7.02 7.04 !ss 131 135 Peak Area M* 1190 1108 Peak Area Ratio 107.4E-2 M- The lattar in" In this column indicates that manual integration a s psrtorm sd on that com pound. Prinl Dale and Tene: 1/18/01 14:58 Page 1 000887 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-265 Page 79 D ata F ile A c q On Sample M ise D:\MSD D A TA \PA B \PA B X -B B A \l-047-l\0471041.D V i a l : 41 3 Jan 2001 00:21 O perator: 10929-GROUP3-F0 In s t : MS7 MVL_________ ___ ______________________________ Amount : ______________0 . 0 0 Io n : 131 (7.017) 0471041.D PABX BBA.M Thu Jan 18 14:58:58 2001 Page 2 000888 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-266 Page 80 Peak Area Report Sample Nam: 10938-GROUP3-F0 notebook Reference: PABX-BBA-1 -047 Data Acquired: 1/3/01 0:41 Operator. Data File Name: 0471042.D Instrument Name: MS7 Data Rie Rath: D:\MSD_0ATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number 42 Instrument Method Name: PABX_BBA Comoound # 1) 2) Comoound Name MVL MVL-d4 Retention Time (Min.) 7.02 7.04 Ion 131 135 Peak Area M* 1604 917 Peak Area Ratio 174.9E-2 M` m The tener "m" In this column Indicates th a t manual Integration was performe d on that compound. Print Date and Time: 1/18/01 14:59 Page 1 000889 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-267 Page 81 y u a iiL ik O L u u u tw * i v o i c * c u > Data File A c q On Sample M ise D:\MSD D A TA \PA B \PA X -8B A \l-047-l\0471042 .D V ia l: 42 3 Ja n 2001 00:41 O perator: 10938-GROUP3-F0 Inst : MS7 I n te g r a tio n P a rs e s : RTEINT. P L a n t M ethod : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL___________________ ____ Io n : 131 (7 .0 1 6 )" , R e s p : 1604 ___________ Amount : 0.00 0471042.D PABX BBA.M Thu Jan 18 14:59:17 2001 Page 2 (J00890 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-268 Page 82 Peak Area Report Sample Name: 10942-GROUP3-FO Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3AJ1 1:00 Operator Data FileName: 0471043.D Instrument Name: MS7 Data File Path: D:\MSD_DATAVPAB\PABX-BBA\1-047-1\ Instrument Vial Number 43 Instrument Method Name: PABX_BBA Compound * 1) 2) Comoound Nam MVL MVL-d4 Retention Tim (Min.) 7.02 7.04 Ion 131 135 Peak Area W 1938 777 Peak Area Ratio 249.4E-2 U ' w The letter "m" In this cahim n indicates thst manual tntsgration was pertorm sd o n that compound. Print Date and Time: 1/1B/01 14:59 Page 1 000891 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-269 Page 83 y u a n tita tio n tteport R eview ed/ Data F ile A cq On Sample M ise D :\M SD D A TA \PA B \PA B X -B B A \1-047-1\0471043.D V i a l : 43 3 J a n 2001 1:00 O perator: 1 0 9 4 2 - GROUPS-F0 In st : MS7 I n t e g r a t i o n Param s: RTEINT.P a n t M eth o d : D:\HPCHEM\l\MKTHODS\PABX_BBA.M (RTE I n t e g r a t o r ) ism:-::: " TIC:04710400 MVL Amount: 0.00 0471043.D PABX_BBA.M Thu Jan 18 14:59:37 2001 Page 2 000892 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-270 Page 84 Peak Area Report Sample Name: 10941-GROUP3-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 1:20 Operator: Data File Name: 0471044.D Instrument Name: MS7 Data File Path: D:\MSD_DATA\PAB\PABX-B8A\1-047-1\ Instrument Vial Number At Instrument Method Name: PABX.BBA Comoound # H 2) Comoound Name MVL MVL-04 Retention Time (Min.) 7.01 7.04 Ion 131 135 Peak Area M* 4575 1656 Peak Area Ratio 275.9E-2 Ma 3 The letter "m" In this column indicates m et manual Integrettori wee performed on that compound. Print Dale and Time: 1/18/01 14:59 Page 1 000893 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-271 Page 85 v u au tjik ab iu u n ep u ic tv* n e v ^ e v e u i Data F ile A c q On Sample M ise D:\MSD D A TA \PA B\PA B X -B B A \l-047-l\0471044.D V i a l : 44 3 Jan 2001 1:20 O perator: 10941-GROOP3-FO Inst : MS7 I n te g r a tio n Pararne: RTEINT.P a n t M e th o d : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL Amount: 0.00 0471044.D PABX BBA.M Thu Jan 18 14:59:57 2001 Page 2 000894 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-272 Page 86 Peak Area Report Sample Name: 10929GROUP3-F1 Notebook Reference: PASX-BBA-1-047 Date Acquired: 1/3A1 1:40 Operator: Data FileName: 0471045.D Data File Path: D:\MSD_DATA'PAB'PABX-B8AVI-047-1\ Instrument Method Name: PABX.BBA kiatiument Name: MS7 Instrument Vial Number: 45 ComDound 1) 2) Comoound Name MVL MVL-d4 Retention Tima IMin.l 7.02 7.04 ton 131 135 Peak Area M* 1254 1183 Peak Area Ratio 106.0E-2 M* > The tettar " n r In this column ktdlcatas that manual In te a iatk ai n o s partorm ad o n that compound. Print Dale and Time: 1/18/01 15:00 Page 1 000995 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-273 Page 87 l. w * asm cm / Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047 -1 \0 4 7 1 0 4 5 . D V ia l: 45 3 J a n 2001 1:40 O perator: 1 0 9 2 9GROUP3- F l Inst : MS7 j Integration Params: RTEINT.P t M eth o d ; D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) I ..... .. 11 1 .... .... I ..... ! 1 1 1I .... r i*t *i , j"` r , .................. ! . . )'. . f I . .- T | I-I 'I 1 I I 1 I ' ' ' ' I r f > ' " H "1 ' "T 6.20 6.30 6.40 S.SO 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7230 7.40 7.50 7.60 7.70 7.BO 7.00 0471045. D PABX BBA.M Thu Jan 18 15:00:16 2001 Page 2 000896 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-274 Page 88 Peak Area Report Sample Name: 10942-GROUP3-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 1:59 Operator: Data FileName: 0471046.D Data Rie Path: D:\MSD_DATA\PAB\PABX-8BA\1-047-1\ Instrument Method Name: PABX_BBA Instrument Name: M S 7 Instrument Vial Number: 46 Comoaund # 1) 2) Comoound Nam MVL MVL-d4 Retention Time IMin.) 7.02 7.CM ton 131 135 Peak Area M* 2074 882 Peak Area Ratio 235.1E-2 M* - The letter `m ' in pile column Indicates thet manual Integration w as performed on that compound. Print Date and Time: 1/18/01 15:00 Page 1 000897 Primedica-Worcester Project Number: PABX-BBA 418-018:P A G E 1-275 Page 89 Data F ile Acq On Sample M ise D:\MSDDATA\PAB\PABX-BBA\1-047-1\0471046.D V i a l : 46 3 Jan 2001 1:5 9 10942-GROP3-F1 O perator: Inst : MS7 MVL _ ___________________________________ Io n : _131 (7 .0 15 ) ; Reap: 2074 Amount :_________0 00 0471046.D PABX BBA.M Thu Jan IB 15:00:36 2001 Page 2 000898 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-276 Page 90 Peak Area Report Sam ple Nam e: 1 X QC HIGH N otebook R eference: PABX-BBA-1-047 Date Acquired: 1/3/01 2:19 Operator. Data File Nam e: 0471010.D Data File Path: D:'WSD_0ATA\PAB\PABX-BBAM-047-1\ Instrument Method Name: PABX_BBA Instrum ent Name: MS7 Instrum ent Vial N um ber. 10 Comoound # h 2) C om o o u nd Nam MVL MVL-04 Retention Time (Min.) 7.02 7.04 ion 131 135 Peak Area M* 1470 1091 Peak Area R at 134.7E-2 IT * The tetter "m" m this colum n Indicates that manual Integration wea performed on that com pound. Print Dale and Time: 1/18/01 14:47 Page 1 000899 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-277 P age 91 y u a u c i.u a k .iO u u- w-*- <ve -u < Data F ile Acq On Sample M ise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471010.D V i a l : 10 3 Jan 2001 2:19 O perator: 1 X QC HIGH In a t : MS7 I n t e g r a t i o n Params: RTEINT.P an t Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL________________________________________________Amount : __ io n : 131 (7.016) : Resp: 1470 Q.J30 0471010.D. PABXJ3BA.M Thu Jan 18 14:47:49 2001 Page 2 000900 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-278 Page 92 Peak Area Report Sample Name: 10945-GROUP4-F0 N otebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 2:39 O p e ra to r: Data Fite Name: 0471047.D Data File Path: O:\MSD_DATA\PAB\PABX-BBA\1-047-1\ instrum ent Method Name: PABX_BBA Instrum ent Name: MS7 tnatrum ent Vial N u m b e r 47 Comoound # V 2) C om pound Name MVL MVL-4 Retantion Time (Min.l 7.02 7.04 ton 131 135 Peak Area M* 6392 505 Peak Area Ratio 126.6E-1 M'm The tetter "m ' In this column mdtcatoa that manual tntaqration a performed on that compound. Print Date and Tune: 1/18/01 15:00 Page 1 000801 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-279 Page 93 Data F ile Acq On Sample Mise i^ uan titatiou n epuii u view eu j D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471047.D V i a l : 47 3 Jan 2001 2:39 O perator: 10945- GROUP4 - FO In st : MS7 MVL _ __ _______ _____________________Amount : T i l, Ion : 1 7016 j 0 00 0471047.D PABX BBA.M Thu Jan IB 15:00:56 2001 Page 2 000902 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-280 Page 94 Peak Area Report Sample Name: 10946-GROUP4-F0 N otebook Reference: PABX-BBA-1 *047 Date A cquired: 1/3/01 2:58 Operator: Data FHe Nam e: 047048.0 Data R ie Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX_BBA Instrument Name: MS7 Instrum ent V ial N um ber. 48 Comoound 1) 2) Comoound Name MVL MVL-4 R e te n tio n Time IMIn.i 7.02 7.04 k>n 131 135 Peak Area M* 9124 909 Peak Area Ratio 100.4E-1 M* The letter "m" In this column Indlc s teo that menual kitegrebon tees performed on thet compound. Preti Dale and Time: 1/18/01 14:44 Page 1 000903 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-281 Page 95 uuantxcacion Keport iw tievxeweuy Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1- 0 4 7 - 1\047048. D V i a l : 48 3 Jan 2001 2:58 O perator: 10 9 4 6 -GROUP4-FO In st : MS7 I n t e g r a t i o n Parama: RTEINT.P an t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) Za,f'm "" ................ .. VC: 047048.D MVL Amount: 0.00 047048.D PABX BBA.M Thu Jan IB 14:44:21 2001 Page 2 000904 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-282 Page 96 Peak Area Report Sample Name: 10947-GROUP4-F0 N otebook Reference: PABX-BBA-1-047 Date A cquired: 1/3/01 3:18 Operator: Data File Name: 0471049.D Data R ie Path: O:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX.BBA Instrum ent Name: MS7 Instrum ent Vial N um ber 49 Comoound # U 2) Compound Nam MVL MVL*d4 R e te n tio n Time M in .) 7.02 7.04 ton 131 135 Peak Area M* 9253 986 Peak Area Ratio 936.5E-2 M' - The lattar `m* In this column Indice!** th a t manual Integration o a a performed a n that compound. Prinl Dale and Tane: 1/18/01 15:01 Page 1 000905 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-283 Page 97 Requit iw* Data F ile Acq On Sample Mise D:\MSD OATA\PAB\PABX-BBA\1-047-1\0471049.D V i a l : 49 3 Jan 2001 3:1B 10947-GROUP4-F0 O p erator: Inst : MS7 I n t e g r a t i o n Param s: RTEINT. P a n t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL________________________________________________Amount : r Io n : 131 (7 .~019 ) t Resp: 9253 0.00 0471049.D PABX BBA.M Thu Jan 18 15:01:16 2001 Page 2 000906 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-284 Page 98 Peak Area Report Sample Name: 10949-GROUP4-F0 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 3:37 O p e ra to r: Data File Neme: 0471050.D Data File Path: O:\MSD_DATA\PAB\PABX-8BA\1-047-1\ Instrument Method Name: PABX_BBA k is tn im e n t Name: MS7 kis tru m e n t Vial N um ber: 50 Comoound # U 2) C om oound Name MVL MVL-4 R e te n tio n Time (Min.) 7.01 7.04 bn 131 135 Peak Area M* 4358 947 Peak Area Ratio 460.2E-2 M* The letter *m* In th is colum n indicele* the! manuel Integration wee performed on th at compound. Print Date and Time: 1/18/01 15:01 Page 1 000907 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-285 Page 99 uuanci t a t i on Keporc lyi' Keviewea; Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047- 1 \ 0 4 710 50 .D V i a l 3 Jan 2001 3:37 O p erator 10 9 4 9 -GR0UP4- F0 In st 50 MS7 k3 I n t e g r a t i o n Params: RTEINT.P l i a n t Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL Amount : 0.00 0471050.D PABX BBA.M Thu Jan 18 15:01:36 2001 Page 2 000908 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-286 Page 100 Peak Area Report Sample Name: 10950-GROUP4-F0 Notebook Reference: PABX-BBA-1-047 Dale A cquired: 1/3/D1 3:57 Operator. Data R ie Name: 0471051.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ Instrument Method Name: PABX^BBA Instrum ent Name: MS7 Instrum ent Vial N u m b e r 51 ComDound * 1) 2) Com ootm d Name MVL MVL-4 R e te n tio n Time fMin.l 7.02 7.04 Ion 131 135 Peek Area M* 17179 1993 Peak Area Ratio B62.0E-2 M* * The tetter "m* in th coturno tndlcstes th et manual tntograden area performed on that compound. P m l Date and Time: 1/18/01 15:01 Page 1 000909 Primedica-Worcester Project Number: PABX-BBA yu an cicatio n Keport u ii Kevreweaj Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471051.D V i a l : 51 3 Jan 2001 3:57 O perator: 1 0 9 5 0 -GROUP4-F0 In st : MS7 I n t e g r a t i o n Params: RTEINT.P an t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) 418-018:PAGE 1-287 Page 101 MVL _______________________________________________Amount :_________0.00 ` "ion: 13 i" " n .0 7 ) : Resp: 17179 135 (7.044) 1993 6.20 6 .30 6.40 6.50 6.60 6.70 6.90 6.90 7.00 7,10 7,20 7.30 7.40 7.50 7.60 7.TO 7.B0 7.90 I 0471051.D PABXBBA.M Thu Jan 18 15:01:56 2001 Page 2 000910 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-288 Page 102 Peak Area Report Sample Name: 10951-GROUP4-F0 Notebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 4:17 O perator Data File Name: 0471052.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ kiatnim ent Method Name: PABX_BBA kiatrum ent Name: MS7 Inatnim ent Vial N u m b e r 52 ConriDound * U 2) Com oound Name MVL MVL-4 R eten tio n Time (Min.) 7.02 7.04 ten 131 135 Peak Area M* 18058 2193 Peak Area R at 823.4E-2 M* The letter *m' in thia column Indlcatae th at manual mtagratlon waa partormed on th at com pound. Print Data and Time: 1/18/01 15:02 Page 1 000911 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-289 Page 103 U u an titatio n Report ea tiv i Review Data F ile Acq On Sample N isc D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471052.D V i a l : 52 3 Jan 2001 4 :17 O perator: 10951-GROUP4-FO Inst : MS7 I n t e g r a t i o n Pararne: RTEINT.P an t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL Amount: 0.00 0471052.D PABX BBA.M Thu Jan IB 15:02:1 2001 Page 2 000312 Primedica-Worcester Project Number: PABX-BBA 418-018 .-PAGE 1-290 Page 104 Peak Area Report Sam ple Name: 10953-GROUP4-F0 N otebook Reference: PABX-BBA-1-047 Date A cquired: 1/34)1 4:36 Operator: Data File Name: 0471053.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX.BBA Instrum ent Name: MS7 Instrum ent Vial N um b e r S3 Comoound # 1) 2) Comoound Nam* MVL MVL-4 R e te n tio n Time (Min.) 7.02 7.04 ton 131 135 Peek Aree M* 12921 1584 Peak Aree Ratio B15.7E-2 M* - The letter "m ` In this colum n Indicates that manual Integration w ee perform ed on m et compound. Print Data and Time: I/1B/01 15:02 Page 1 000913 Primedica-Worcester Project Number: PABX-BBA 4 1 8 -0 1 8:PA G E 1-291 Page 105 U U O U U i L A L i U l KffpUll fttVieWttJJ Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471053.D V i a l : 53 3 Jan 2001 4:36 O p erator: 1 0 9 5 3 -GROUP4-F0 Inst : MS7 I n t e g r a t i o n Params: RTEINT. P a n t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL_______________________________________________Amount :________ 0.00 Io n : 1 3 l " ( 7 `. 015) ! Reap: 12921 0471053.D PABX BBA.M Thu Jan 18 15:02:35 2001 Page 2 000914 Primedica-Worcester Project Number: PABX-BBA 418-018:P A G E 1-292 Page 106 Peak Area Report Sam ple Name: 10954-GROUP4-F0 N otebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3A01 4:56 Operator. Data File Name: 0471054.0 Data File Path: D:\MSD_DATA1PAB\PABX-BBA\1-047-1\ Instrum ent M ethod Name: PABX_BBA Instrument Name: MS7 Instrum ent Vial Number. 54 Comoound # U 2) Comoound Nam MVL MVL-d4 R e te n tio n Time MIn.l 7.02 7.04 ton 131 135 Peek Area M* 11975 1860 Peek Arne Ratio 637.0E-2 M * - The letter 'm 'lf i this column Indicates that menus! integration teas performed on that compound. Print Date and Time: 1/18/01 15:02 Page 1 000915 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-293 Page 107 U U d U L ^ LO U X U ll K c p o i. L lu x n c v x c d e u i Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-0 4 7 - 1 \ 0471054.D V i a l : 54 3 Jan 2001 4:56 O perator: 1 0 9 5 4 -GR0UP4-FQ Inst : MS7 I n t e g r a t i o n Pararne: RTEINT.P a n t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) TIC: 0471054D MVL ____________________________________ i Ion: 131 (7.017) Amount :________ 0.00 0471054.D PABX BBA.M Thu Jan 18 15:02:55 2001 Page 2 000916 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-294 Page 108 Peak Area Report Sample Name: 10945-GROUP4-F1 Notebook Reference: PABX-0BA1-O47 Date A cquired: 1/3/01 5:15 O p e ra to r. Data Rte Name: 0471055.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX.BBA Instrum ent Name: MS7 kistrum ent Vial Number: 55 Comoound # 1) 2) C om oound Name MVL MVL-4 Retention Tim e (Min.) 7.01 7.04 Ion 131 135 Peak Area M* 285656 1720 Peak Area Ratio 166.2E+0 M* mThe Iettar *m * in th i* column indicates that manual integration w m perform ed on that compound. Print Date and Time: 1/18/01 15:03 Page 1 000B 17 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-295 Page 109 W UaIlt.luat..LOn ttepuX L vyi RB V itw cuJ Data F ile Acq On Sample Mise D:\MSD nATA\PAB\PABX-BBA\l-047-l\0471055.D V i a l : 55 3 Jan 2001 5:15 O perator: 10945-GROOP4-F1 In st : MS7 I n t e g r a t i o n Params: RTEINT.P a n t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL _ _ ____ __ _________ __________ Amount :______ 0.00 : 'ion: 131 "(7.013) ' R esp: 2B58S6 MVTj-d4 Amount: 0.00 0471055.D PABX BBA.M Thu Jan IS 15:03:15 .1001 Page 2 000918 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1-296 Page 110 Peak Area Report Sample Name: 10946-GROUP4-F1 N otebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 5:35 O perator Data File Name: 0471056.D Data File Path: D:\MSD_DATA\PABtPABX-BBA\1-047-1\ Instrum ent Method Name: PABX_8BA Instru m e n t Name: MS7 b is tru m e n t VUri N u m b e r 56 Comoound # 1) 2) C om pound Name MVL MVL-d4 R e te n tio n Time IMIn.l 7.02 7.04 Ion 131 135 Peak Area M' 380925 2454 Peak Area Ratio 155.2E+0 M * at The letter *m * In thle column Indicetee that m anual Integration wee performed on that compound. Phot Date and Time: 1/18/01 15.03 Page 1 000919 Primedica-Worcester Project Number: PABX-BBA 418-018:P A G E 1-297 P age 111 U u a u u iu a u iu u ficpwi c v* A e ie e u i Data F ile Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\1-047-1\0471056.D V i a l : 56 3 Jan 2001 5:35 O perator: 10946-GROUP4- F l In st : MSI I n t e g r a t i o n Param s: RTEINT.P 0 * " t Method : D:\HPCHEMU\METHODS\PABX_BBA.M (BTE I n t e g r a t o r ) MVL M'rr,-d4 Amount: 0.00 Amount: 0.00 0471056.D PABX BBA.M Thu Jan 18 15:03:35 2001 Page 2 000920 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-298 Page 112 Peak Area Report Sample Name: 10947-GROUP4-F1 N otebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 5:55 O perator Data R e Name: 0471057.0 Data Fite Path: D:\MSD_OATA\PAB\PABX-BBA\1-047-1\ Instrum ent M ethod Name: PABX_BBA tastrum ent Name: MS7 Instrum ent Vial N u m b e r 57 Comoound 1 U 2) Com oound Name MVL MVL-4 R e te n tio n Time (Min.) 7.01 7.04 ten 131 135 Peek Area j 180712 1824 Peek Aree Ratio 990.7E-1 M* * The letter am * in this column Indicata* that manual Integration mi performed on that compound. Prmt Date and Time: 1/18/01 15:03 Page 1 000921 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-299 Page 113 Data F ile Acq On Sample Mise u u a n tita tio n Report tu i Reviewed/ D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471057 .D V i a l : 57 3 Jan 2001 5:55 10947-GR0CJP4- F l O p erator: In st : MS7 MVL_ __ _ .00^_________________________________Amountj________0 i Ion: 13 17.012) ; Reap: 1B0712 MVL-d4 Amount : 0.00 0471057.D PABX BBA.M Thu Jan 18 15:03:55 2001 Page 2 000922 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-300 Page 114 Peak Area Report Sample Name: 10949-GROUP4-F1 Notebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 6:14 Operator. Data FUe Name: 0471056.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\t-047-l\ instrum ent Method Name: PABX.BBA Instrument Name: MS7 kistm m e nt Vial N um ber. 58 Comnound * 1) 2) C om oound Name MVL MVL-04 R e te n tio n Tim e fMin.1 7.02 7.04 SQ 131 135 Peak Area M230169 1560 Peak Area Ratio 147.5E+0 M* * The le tte r' m" In this column indicates that manual Integration was performed on thet compound. Print Date and Time: 1/18/01 15:04 Page 1 000923 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-301 Page 115 y u a n t it a t io n Report (U't Reviewed/ Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1 \04 71058 .D V i a l : 58 3 Jan 2001 6:14 10949-GROUP4-F1 O perator: In st : MS7 I n t e g r a t i o n Params: RTEINT.P Lant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE I n t e g r a t o r ) MVL_______________________________________ __ Ion: 131 (7.017) R esp: 230169 Amoun t : _0^00 0471058.D PABX BBA.M Thu Jan 18 15:04:15 2001 Page 2 000924 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-302 Page 116 Peak Area Report Sam ple Name: 2 X OC LOW N otebook Reference: PABX-BBA-1-047 Date A cquired: 1/3701 6:34 Operator: Data File Name: 0471015.0 Data Fife Path: D:\MSD_DATA\PAB1PABX-BBA\1-047-1\ Instrum ent Method Name: PABX_BBA Instrum ent Name: MS7 Instrum ent Vial Num ber. 15 Comoound # K 2) Com pound Name MVL MVL-d4 R e te n tio n Time /Min.) 7.02 7.04 Ion 131 135 Peek Area M* 502 1343 Peak Area Ratio 373.8E-3 M* a The tettar -m * In this column indicates ttua manual kuayratiun m n performed an that compound Print Date and Time: 1/18/01 14:49 Page 1 000925 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-303 Page 117 Q uantita txo n Report (QT Reviewed) Data F ile Acq On Sample M ise D:\MSD DATA\PAB\PABX-BBA\1 - 0 4 7 - 1 \ 0 4 7 1 0 1 5 . D V i a l : IS 3 Jan 2001 f i : 34 O p erator: 2 X QC LOW Inst : MS7 I n t e g r a t i o n Paratns : RTEINT.P m an t Method : D: \HPCHEM\ 1 \METHODS\PABX_BBA. M (RTE I n t e g r a t o r ) MVL_____________________ _ I " io n : "13 (7.016) Resp : 502 __________________Amount :_________0.00 ________________________ A . .. k 620 620 6.40 M VL-d4 f ^ n : 135 (7.043) ^ sp : 1343 6.S0 6.60 6.70 6.80 6.90 7 00 Amount: 7.10 720 0.00 730 7.40 7.50 7.60 7.70 7.80 7.90 j 'i A 720 I ' 1 1 ' I ' ' ' I ' ' ' 1 I ' ' ' ' I ' r ' ' I ' ' ' ' ' ' ' ' ' I ' ' ' ' I ' ' ' ' F ' ' ' ' I ' ' ' ' I ' ' ' ' 1 ' ' ' ' I ' ' ' ' I ' ' I ' ' ' I ' ' ' ' l ' 6.20 6.30 6.40 6.50 6.60 6.7D 6.80 6.90 7.00 7.10 720 7.40 7.50 7.60 7.70 7.80 7.90 0471015.D PABX BBA.M Thu Jan 18 14:49:28 2001 Page 2 000926 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-304 Page 118 Peak Area Report Sample Name: 10950-GROUP4-F1 Notebook Reference: PABX-BBA-1-047 Data A cquired: 1/3/01 6:54 O perator Data File Name: 0471059.D Data File Path: D:\MSD_DATA\PAB\PABX-BBAM-047-1\ instrum ent Method Name: PABX_B8A tnatrum ent Name: MS7 Inatrum ent Vial N um ber: 59 Comoound * 1) 2) Com oound Name mvl MVL-04 R e te n tio n tecTime iMin.J Peek Area M' 7.01 131 298676 7.04 135 2623 Peak Area Ratio 113.9E+0 *f* * The letter ` m* In m ie colum n inrjlcetee that manuel integration ma performed on that com pound Print Date and Time: 1/18/01 15:04 Page 1 000927 Priinedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-305 Page 119 Q u a n tita tio n Report (QT Reviewed) Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-B B A \l-0 4 7-l\D 4 710 59 .D V i a l : 59 3 Jan 2001 6:54 O perator: 109S0-GROUF4-F1 In st : MS7 I n t e g r a t i o n Pararas: RTEINT.P a n t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M IRTE I n t e g r a t o r ) TIC: 0471059.D MVL_ ________________________________________Amount :_______ 0.00 ' 'io n ':"3 1 "(7.0 15) : Resp: 298676 0471059.D PABX_BBA.M Thu Jan 18 15:04:35 2001 Page 2 000928 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-306 Page 120 Peak Area Report Sample N a n : 109S1-GROUP4-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 7:13 O p e ra to r Data File Name: 0471060.D Data File Path: D:\MS0_DATA\PA8\PABX-BBA\1>047-1\ Instrument Method Name: PABX-BBA Instru m e n t Name: MS7 In s tru m e n t V ial N u m b e r 60 ComDOund * 1) 2) C om oound Name MVL MVL-d4 R e te n tio n Tima IMln.) 7.02 7.04 tea 131 135 Peak A n a M* 292458 2139 Peak Area Ratio 136.7E+0 M* mThe latter *m a In this eokim n Indicate! that manual Integration w as perform ed on that compound. Print Date and Time: 1/18/01 15:04 Page 1 000929 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-307 P age 121 Quantitation Report IUT Reviewed) Data File Acq On Sample Mise D:\MSD DATAIPAB\PABX-B8A\1-047-1\0471060.D Vial: 60 3 Jan 2001 7:13 Operator: 10951-GROP4-F1 Inst : MS7 l', Integration Params: RTEINT.P tant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M RTE Integrator) MVL__________ ______________________ Amount : Ion: *131 (7.017) ; Resp: 292458 0 -00 0471060.D PABX BBA.M Thu Jan 18 15:04:54 2001 Page 2 000330 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-308 Page 122 Peak Area Report Sam ple Name: 10953-GROUP4-F1 Notebook Reference: PABX-B8A-1-047 Dale A cq u ire d : 1/3A31 7:33 O p e ra to r: Data File Name: 0471061.D Data File Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX_BBA Instrum ent Name: MS7 Instrum ent Vial Num ber. 61 Comoound # 1) 2) C om pound Name MVL MVL-4 R a te n tio n Time fMln.j 7.02 7.04 ton 131 135 Peak Area M' 241386 2064 Peak Area Ratio 115.66*0 M' The letter "m" m this column Indlcatea that manual imogretkm m e performed on th at compound. Print Date and Time: 1/18701 15:05 Page 1 000931 Primcdica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-309 P age 123 Quantitation Report IQT Reviewed! Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471061.D Vial: SI 3 Jan 2001 7:33 Operator: 10953-GROUP4-Fl Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) T1C:0471061D M V L ____ __________________ __________ Amount :______ 0.00 : "n: 131 (7.016) IResp: 2413S6 0471061.D PABX BBA.M Thu Jan IB 15:05:14 2001 Page 2 000932 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-310 Page 124 Peak Area Report Sample Hama: 10954-GROUP4-F1 N otebook Reterance: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 7:53 O p e ra to r Date File Name: 0471062.D Date File Patti: 0 :'iMSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX.BBA Instrument Name: MS7 Instrument Vial N u m b e r 62 Comoound # 1) 2) Comoound Name MVL MVL-d4 R e te n tio n Time (Min.) 7.02 7.04 ten 131 135 Peak Area M* 226425 1644 Peak Area Ratio 122.8E+0 M* > The tener 'm ' In M e column indicates that m anual Integration eras performed on that com pound. Print Dale and Time: 1/18/01 15:05 Page 1 000933 Primedica-Worcester Project Number: PABX-BBA 418-Q18:PAGE 1-311 Page 125 Uuancication Report ivi neviemui Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\l-047-l\0471062.D Vial: 62 3 Jan 2001 7:53 Operator: 10954-GR0UP4-Fl Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) TIC: 0471062.0 MVL Amount: 0.00 0471062.0 PABX BBA.M Thu Jam IB 1S:05:34 2001 Page 2 000934 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-312 Page 126 Peak Area Report Sample Nam: 10943-GROUP4-F0 N otebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 8:12 O p e ra to r: Data File Name: 0471063.D Data R ie Path: O:\MSO_DATA\PAB\PABX-BBA\1 -047-1\ Instrument Method Name: PABX_BBA Instrum ent Name: MS7 k u tram ent Vial Number. 63 Comoound # U 2) C om oound Name MVL MVL-4 Retention Time (Min.l 7.02 7.04 Ion 131 135 Peak Area M* 1753 1939 Peak Area Ratio 904. IE-3 M* * The letter *m" tn this column Indicates th at manual integration area performed on th at com pound. Print Date and Tette: 1/18/01 15:05 Page 1 000935 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-313 Page 127 UUdnU.LdLlUII Kdpux L WI Rdvxeweui Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471063 .D Vial: 63 3 Jan 2001 8:12 Operator: 10943-GROUP4-FO Inst : MS7 Integration Parants: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL __________________________________Amount : '" l o h : `l31 (7.016) _0.00 0471063.D PABX BBA.M Thu Jan 18 15:05:53 2001 Page 2 000936 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-314 Page 128 Peak Area Report Sample Name: 10948-GROUP4-F0 Notebook Reference: PABX-BBA-1 -047 Date A cq u ire d : 1/3/01 8:32 O p e ra to r: Data File Name: 0471064.D Data File Path: D:\MSCLDATA\PAB\PABX-BBA11-047-1\ kiatium ent Method Name: PABX_BBA Instrument Name: MS7 Instrum ent Vial N u m b e r 64 Cofnoound # 1) 2) ComDOund Name MVL MVL-cW R eten tio n Time (Min.i 7.02 7.04 131 135 Peak Area M* 2188 1996 Peak Area Ratio 109.6E-2 M* The letter V in this column Indicates that manual Integra tion area performed on that co mpound. Print Oate and Time: 1/18/01 15:06 Page 1 000937 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-315 Page 129 uuantltation Report iv neieeui Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\04 71064 -D Vial: 64 3 Jan 2001 8:32 Operator: 10948-GROUP4-FO Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) i.ifijB .c- " 710:047106433* MVL_________ _____________________ ____ Amount :________0.00 I Ion: 131 (7*017) 0471064.D PABX BBA.M Thu Jan 18 15:06:13 2001 Page 2 000938 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-316 Page 130 Peak Area Report Sam ple Name: 10952-GROUP4-F0 N otebook Reference: PABX-0BA-1-O47 Oete A cq u ire d : 1/3/01 8:52 O p e ra to r: Data File Name: 0471065.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX_BBA Instru m e n t Nam e: MS7 tostrum ent Vial N u m b e r 65 Comoound # 1) 2) Comoound Name MVL MVL*d4 R ale n ti on Time IMIn.l 7.01 7.04 ton 131 135 Peak Area M* 1778 2617 Peak Area Ratio 679.4E-3 M* * The Iettar *m* in this column hxHc*tss m at manual integration was performed on that compound. Pnnt Date and Time: 1/18A)1 15:06 Page 1 000939 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-317 P age 131 V U a n C IC ttU U U Ite p u iL t v * **=v i c n e u ; Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471065.D Vial: SS 3 Jan 2001 8:52 Operator: 10952-GROUP4-FO Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator! 0471065.D PABX BBA.M Thu Jan 18 15:06:32 2001 Page 2 000940 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-318 Page 132 Peak Area Report Sample Nam: 10955-GROUP4-FO N otebook Reference: PABX-BBA-1 -047 Data Acquired: 1/3/01 9:11 Operator: Data File Name: 0471066.0 Data File Path: O:\MSD_DATA\PAB\PABX-BBA\1-Q47-1\ Instrum ent Method Name: PABX_BBA Instrum ent Name: MS7 Instrum ent V ial Num ber. 66 Comoound 1) 2) Comoound Nam MVL MVL-cM R e te n tio n Time (Min.i 7.02 7.04 ion 131 135 Peak Aree M' 3322 2412 Peek Area Ratio 137.7E-2 M` - The le tte r'm ' In thte column Indicates that manual integration wee pertarm ed on that compound. Prinl Date and Tme: 1/1B/01 15:06 Page 1 000941 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-319 Page 133 Daca File Acq On Sample Mise D:\MSD OATA\PAB\PABX-8BA\1-047-1\0471066.D Vial: 66 3 Jan 2001 9:11 Operator 10955-GRODP4-F0 Inst MS 7 ; Integration Params: RTEINT.P tant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) M V L _________ ____ __ _______ _________ Amount : _____ 0^0 0 "ln: 13i <7~.08) *' 0471066.D PABX BBA.M Thu Jan 18 15:06:52 2001 Page 2 000942 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-320 Page 134 Peak Area Report Sample Name: 10956-GROUP4-F0 Notebook Reference: PABX-BBA-1*047 Date A c q u ire d : 1/3/01 9:31 O p e ra to r: Data File Name: 0471067.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ Instrument Method Name: PABX.BBA kistrum ent Name: MS7 Instrum ent V ial N um ber: 67 ComDOund * 1) 2) ComDound N airn MVL MVL-d4 R e te n tio n Time fMin.) 7.01 7.04 Ion 131 135 Peek Area M* 357223 2339 Peak Aree Ratio 152.7E+0 M* * The letter *m" In this colum n indlcatee that manual Integration w et performed on th a t com pound. Print Date and Time: 1/18/01 15:07 Page 1 000943 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-321 Page 135 VU d.uuj.uat.xuu ite^iuxL vyi n e u e n e u j Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471067.D Vial: 67 3 Jan 2001 9:31 Operator: 10956-GROUP4-F0 Inst : MS7 Integration Pararas: RTEINT.P ant Method : D :\HPCHEMU\METHODS\PABX_BBA.M (RTE Integrator) MVL____________________ ________ : Ion: 131 (7.014) : Resp: 357223 Amount:______ 0 .00 MVL-d4 Amount: 0.00 0471067.D PABX BBA.M Thu Jan 18 15:07:11 2001 Page 2 000944 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-322 Page 136 Peak Area Report Sam ple Nam: 10944-GROUP4-F0 N otebook Reference: PA8X-BBA-1-047 Date Acquired: 1/3/01 9:51 Operator: Data File Name: 0471068.D Data File Patti: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX_BBA Instrum ent Name: MS7 Instrum ent Vial Number. 66 ComDound f U 2) Compound Name MVL MVL-d4 R e te n tio n Time Otfln.i 7.02 7.04 ion 131 135 Peak Area M* 2815 2414 Peak Area Ratio 116.6E-2 M" - The letter ` m* In this column kidlcstsi that msnusl Integration woe perlorm ed on thst compound. Print Date and Time: 1/18/01 15:07 Page 1 000945 Prixnedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-323 Page 137 g u a itv .^ i.ai> ^ u it l. w * n c t i c c u i Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471068.D Vial: 68 3 Jan 2001 9:51 Operator: 10944-GR0UP4-FO Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL___________ _________________________ Amount :______ 0.00 Ion: 131 (7.016) Resp: 2815 0471060.D PABX BBA.M Thu Jan 18 15:07:31 2001 Page 2 000946 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-324 Page 138 Peak Area Report Sampla Name: 11029-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 10:11 Operator. Data Rie Name: 0471069.D Data Rie Path: D:\MSD_DATA\PAB\PABX-8BA\1-047-1\ Instrument Method Nsme: PABX_BBA Instrument Name: MS7 Instrument Vial Number 69 Comoound # 1) 2) Comoound Name MVL MVL*d4 Retention Time (Min.l 7.02 7.04 {on 131 135 Peak Area M* 781 1833 Peak Area Ratio 426.1E-3 M* The tetter *m" in this column ndicetea that manual integration ease performed on that compound. Print Date and Time: 1/16/01 15:07 Page 1 000947 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-325 Page 139 yuantxtation Report Kevxeweaj Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471069.D Vial; 69 3 Jan 2001 10:11 Operator: 11029-GR0UP11-FO Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) MVL ______ _______ __________________ Amount ;______ 0.00 Ion: "13 "(7.016) ;Resp: 781 0471069.D PABX BBA.M Thu Jan IB 15:07:51 2001 Page 2 000948 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-326 Page 140 Peak Area Report Sample Name: 11031-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Dale Acquired: 1/3AJ1 10:31 Operator Data Rie Name: 0471070.D Instrument Name: MS7 Data Rie Path: D:\MSD_DATA1PAB\PABX-BBA\1-047-1\ Instrument Vial Number. 70 Instrument Mathod Name: PABX.BBA Comoound i) 2) Comoound Name MVL MVL-cW Retention Time (Min.l 7.02 7.04 Ion 131 135 Peak Area NT 710 1713 Peak Area Ratio 414.5E-3 M* The letter "m * In this column kidicstes Ourt manual Integration teas performed on that compound. Print Date and Time: 1/18/01 15:08 Page 1 000949 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-327 Page 141 Quantitation Report lui Keviewem Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471070 .D Vial: 70 3 Jan 2001 10:31 Operator: 11031-GR0P11-FQ Inst : MS7 i3 Integration Params: RTEINT.P tant Method : D:\HPCHEM\l\KETHODS\PABXJBBA.M (RTE Integrator) MVL :~Ton: 131 (7.017) ! Resp: 710 _____Amount :______ 0 00 0471070.D PABX BBA.M Thu Jan 18 15:08:10 2001 Page 2 000950 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-328 Page 142 Peak Area Report Sample Name: 2 X Q C MID Notebook Reference: PA8X-BBA-1-047 Date Acquired: 1/3/01 10:50 Operator: Oata FiteName: 047101^.0 InstrumentName: M S 7 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number: 13 Instrument Method Name: PABX_BBA Comoound # 1) 2) Comoound Nam MVL MVL-d4 Retention Time (Mln.l 7.02 7.04 Ion 131 135 Peek Area M* 737 1584 Peak Area Ratio 465.3E-3 M* The letter "m* in this column indicate! that manual krtagration wms performed on that compound. Pnnl Date and Time: 1/18/01 14:48 Page 1 00951 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-329 Page 143 uuancicacion Report. lux neieeu/ Data File Acq On Sample Mise D:\MSD DATA\ PAB\PABX-BBA\1- Q47-1\0471013.D Vial: 13 3 Jan 2001 10:50 Operator: 2 X OC MIU Inst : MS7 Integration Params: RTEINT.P ant Method : D :\HPCHEM\1\METHODS\PABX_BBA.M (RTE Integrator) MVL_________ ______ __ . _________ ______Amount :______ 0 .00 Ion: 131 P". 01S) 0471013.D PABX BBA.M Thu Jan IB 14:48:48 2001 Page 2 000352 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-330 Page 144 Peak Area Report Sample Name: 11032-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 11:10 Operator Data Rie Name: 0471071 .D Data Rie Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX-BBA Instrument Name: MS7 instrument Vial Number: 71 Compound # i> 2) ComDOund Name MVL MVL-44 Retention Time (Min.) 7.02 7.04 km 131 135 Peak Area M* 2BS 1766 Peak Area Ratio 161.4E-3 M* * The letter "m` In thia column indicates that manual Integration was portormad on M at compound. Print Date and Time: 1/18/01 15:08 Page 1 000953 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-331 Page 145 Data File Acq On Sample Mise Uuantitation sport (UT Kevieweaj D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471071.D Vial: 71 3 Jan 2001 11:10 Operator: 11032-GBOUP11-F0 Inst : MS7 MVL____________ ____________________ Amount :___ Ion: 131 (770151" Resp: 285 0.00 0471071,D PABX_BBA.M Thu Jan 18 15:08:30 2001 Page 2 000954 Primedica-Worcester Project Number: PABX-BBA 418-018.-PAGE 1-332 Page 146 Peak Area Report Sample Name: 11033-GROUP11-F0 Notebook Reference: PA8X-BBA-1-047 Date Acquired: 1/3/01 11:30 Operator: Data Rie Name: 0471072.0 fcistniment Name: MS7 Data File Patti:O:\MSD_DATA\PAB\PABX-BBA\1 -047-11 Instrument Vial Number 72 Instrument Method Name: PABX_B8A Compound 4 1) 2) C o m D on nd Name MVL MVl-64 Retention Time (Min.) 7.01 7.04 ton 131 135 Peak Area M* 481 2422 Peak Area Ratio 198.6B-3 M - rnThe letter *m * In IM a column Indicete* that manual Integration araa performed on that compound. Print Date and Tane: 1/18/01 15:08 Page 1 000955 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-333 Page 147 yuauhikakiuti kW1 Keie*eu/ Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471072.D Vial: 72 3 Jan 2001 11:30 Operator: 11033-GR0UP11-FO Inst : MS7 Integration Parants : RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL Ion: 131 (7.012) Amount:______ 0.00 135 <7.040) 2422 620 6.30 6.40 --- --^ ' I ...... ..... "T" I ,r 6.60 6.70 6.60 I ' ' * l ' ' T' ' -n~-' - - I ' I 1 1 <" 7.00 7.10 720 7.30 7.40 7.50 7.60 7.70 7.90 0471072.D PABX BBA.M Thu Jan 18 15:08:49 2001 Page 2 000956 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-334 Page 148 Peak Area Report Sample Nam-. 11034-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 11:50 Operator Data File Name: 0471073.0 Initniment Name: MS7 Data Rie Path: D:\MSD_0ATA\PAB1PABX-BBAV1-047-1\ Inatniment Vial Number: 73 Instrument Method Name: PABX.BBA Compound 1) 2) ComDound Name MVL MVL-44 Retention Time (Min.1 7.01 7.04 Ion 131 135 Peek Aree M' 12941 2613 Peak Area Ratio 49S.3E-2 M* a The letter ' m" In this cokimn IndlcatM that manual Integration m a perform ed on that compound. Print Date and Time: 1/18/01 15:09 Page 1 000957 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-335 Page 149 yuar.cicac.ian Report w i nevieweai Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\ 0471073.D Vial: 73 3 Jan 2001 11:50 Operator: 11034-GROUF11-F0 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) TIC:471073.D MVL Amount: 0.00 0471073.D PABX BBA.M Thu Jan 18 15:09:09 2001 Page 2 000958 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-336 Page 150 Peak Area Report Sample Nam: 11035-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 12:10 Operator: Data FileName: 0471074.D Data FilePath: 0:1MS0_DATA1PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX.BBA Instrument Name: MS7 kistrument Vial Number 74 Comoound # 1) 2) Comoound Nam MVL MVL-4 Retention Time (Min.) 7.02 7.04 ton 131 135 Peak Area M* 647 2448 Peak Area Ratio 264.3E-3 M' The tener "m" Inthiscokimn indicatesthatmanual Integrationmas parformad on thatcompound. Print Date and Tine: 1/18/01 15:09 Page 1 000959 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-338 Page 152 Peak Area Report Sample Name: 11036-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 12:30 Operator. Data File Name: 0471075.0 Instrument Name: MS7 Data File Path: D:\MS0_0ATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number: 75 Instrument Method Name: PABX_BBA Compound # 1) 2) Compound Name MVl MVL-d4 Retention Time (Min.) 7.02 7.04 ton 131 135 Peak Area M* 526 1566 Paak Area Ratio 335.9E-3 M* a The letter*m* Inthis column indicate*met menual Inlegation wee pertormed on thatcompound. Print O ale and Time: 1/ 18/01 15:09 Page 1 000961 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-339 Page 153 uuancication Keporc \yi neviewea; Data File Acq On Sample Mise D:\MSD DATA\ PAB\PABX-BBA\1 -047 -1\ 0471075 .D Vial: 75 3 Jan 2001 12:30 Operator: 1103 6-GROUP11-F0 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\KETHODS\PABX_BBA.M (RTE Integrator) .nemes '' ' TIC:710710' 0471075.D PABX BBA.M Thu Jan 18 15:09:49 2001 Page 2 000962 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-340 Page 154 Peak Area Report Sample Name: 11037-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 12:50 Operator Data Rie Name: 0471076.0 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-U Instrument Method Name: PABX.BBA Instrument Name: MS7 hstrument Vial Number: 76 ComDound 1) 2) Comoound Name MVL MVL-4 Retention Time IMIn.l 7.02 7.04 Ion 131 135 Peak Area M* 542 2016 Peak Area Ratio 268.8E-3 M ** The letter *m * in thie colum n Indleatet ttw manual Integration w ee perform ed on mm compound. * Print Dale and Time: 1/18/01 15:10 Page 1 000963 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-341 Page 155 Uuantitation Report lU'i Kevieweaj Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\1-047-1\0471076.D Vial: 76 3 Jan 2001 12:50 11037-GROUP11-F0 Operator: Inst : MS7 kS Integration Params: RTEINT.P Lant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator! MVL__________ _________________ "ion: 131 {7 7IS) Amount : _ __ 0.0_0 0471076.D PABX BBA.M Thu Jan IB 15:10:09 2001 Page 2 000964 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-342 Page 156 Peak Area Report Sample Name: 11038-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 13:10 Operator Data File Name: 0471077.0 tnatrument Name: MS7 Data File Path: D:1MSD_DATA1PAB1PABX-BBA11 -047-11 Instrument Vial Number 77 Instrument Method Name: PABX_BBA Comoound 1 1) 2) Comoound Name MVL MVL-cM Retention Time (Min.) 7.02 7.04 ton 131 135 Peak Area M" 602 2007 Peak Area Ratio 300.0E-3 U ' a The tetter "m" in this coturni tndlcatee that manual Integration was performed on that compound. Print Date and Time: 1/1B/01 15:10 Page 1 000965 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-343 Page 157 quantitation Keport Kevieweuj Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471077 .D Vial 3 Jan 2001 13:10 11038"GROP11-F0 Operator Inst MS7 G integration Params : RTEINT.P |uant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL _ _ _____ __________________ Amountj_____ 0.00 . on: 131 p.o'lfif IResp: 602 0471077.D PABX BBA.M Thu Jan IB 15:10:29 2001 Page 2 000966 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-344 Page 158 Peak Area Report Sample Name: 11029-GROUP11-F1 Notebook Reference: PABX'BBA-1-047 Date Acquired: 1/3/01 13.30 Operator Data Rie Name: 0471078.D Instrument Name: MS7 Data File Path: D:\MSD_0ATA\PAB\PABX-BBA\1 -047-1\ Ustrument Vial Number 78 Instrument Method Name: PABX_BBA Comoound # 1) 2) Comoound Nam* MVL MVL*d4 Retention Time (Min.) 7.02 7.04 Ion 131 135 Peak Area M* 189 1448 Peak Area Ratio 130.5E-3 M* * The tetter V in this column indicate* m et manual Integration was perform ed on that compound. Print Date and Time: 1/18701 15:10 Page 1 000967 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-345 Page 159 Uuantitation Keport lU'i Kevieweaj Data File Acq On Sample Mise D:\MSD DATA\PAB\PAB.X-BBA\1 -04 7-1 \Q4 71078 .D Vial: 78 3 Jan 2001 13:30 Operator: 11029-GROUP11-F1 Inst : MS7 1 Integration Parants: RTEINT.P kuant Method : O:\HPCHEM\l\METHODS\PABX BBA. iOTl? TIC:047107B.D MVL______________________________ _ ion: 131 (7.016) Resp: 189 ._ J iu n o u n t :________ 0 . 0 0 0471078.D PABX BBA.M Thu Jan 18 15:10:48 2001 Page 2 000968 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-346 Page 160 Peak Area Report Sample Name: 11031-GROUP11-F1 Notebook Reference: PA8X-BBA-1-047 Date Acquired: 113/01 13:49 Operator Data FileName: 0471079.D Instrument Name: MS7 Data FHe Path: D:\MSD_DATA\PAB\PABX-8BA\1 -047-1\ Instrument Vial Number. 79 Instrument Method Name: PABXJ3BA ContDound * 1) 2) ComDOund Name MVL MVL-4 Retention Time fMin.l 7.02 7.04 Ion 131 135 Peek Aree M* 146 1617 Peak Area Ratio 902.9E-4 M ` - The letter *m * In this column Indicates that manual intagration teas performed on that compound. Print Date and Time: 1/18/01 15:10 Page 1 000969 Primedica-Worcester Project Number: PABX-BBA 418-018 .'PAGE 1-347 Page 161 Data File Acq On Sample Mise Quantitation Report IQT Reviewed! D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471079.D Vial: 79 3 Jan 2001 13:49 Operator: 11031-GR0UP11-Fl Inst : MS7 MVL _ __________ _____________ Amount :______ 0.00 7 T o n f is i" (7.019) I Resp: 146 0471079 .D PABX BBA.M Thu Jan 18 15:11:08 2001 Page 2 000970 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-348 Page 162 Peak Area Report Sample Name: 11033-GROUP11-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 14:09 Operator Data File Name: 0471080.D Instrument Name: MS7 Data File Path: D:'MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Number 80 Instrument Method Name: PABX_BBA ComDound # U 2) Compound Name MVL MVL-d4 Retention Time (Min.) 7.02 7.04 ten 131 135 Peak Arsa M* 293 1957 Peek Area Ratio 149.7E-3 M* * The letter "m " In th is colum n indicates th a t menuet integration ma partormed on that compound. Print Dale and Time: 1/18/01 15:11 Page 1 000971 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-337 Page 151 UUantiULlUU Repua. IWI nevieweui Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471074.D Vial: 74 3 Jan 2001 12:10 Operator: 11035-GROUP11-F0 Inst : MS7 Integration Params: RTEINT.P ant Method : D :\HPCHEM\ l\MSTHODS\ PABX_BBA.M IRTE Integrator) MVL Amount: 0.00 0471074.D PABX BBA.M Thu Jan 18 15:09:29 2001 Page 2 000960 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-349 Page 163 Data File Acq On Sample Mise Quantitation Report iyi' Reviewed) D:\MSD DATA\ PAB\ PABX-BBA\ 1-04 7 -1\ 0471080 .D Vial: 80 3 Jan 2001 14:09 Operator: 11033-GROUP11-FI Inst : MS7 MVL ___________________________________ Amount :______ 0 .00 Ion:' 131 <7 J016 ) i Reap: 293 0471080.D PABX BBA.M Thu Jan 18 15:11:28 2001 Page 2 000972 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-350 Page 164 Peak Area Report Sample Name: 11036-GROUP11-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 14:29 Operator Data File Name: 0471081.D kistrumcnt Name: MS7 Data File Path: D:\MS0_0ATA'PAB'PABX-BBAM-047-1\ Inatrumenl Vial Number 81 Instrument Method Name: PABX_BBA Comoound f 1) 2) Compound Nam MVL MVL-d4 Retention Time iMIn.) 7.02 7.04 ton 131 135 Peak Area M* 260 1943 Peak Area Ratio 133.8E-3 M* - The letter"m" m thiscolumn Indicatesthatmenus! integrationwas performed on thatcompound. Print Date and Time: 1/18/01 15:11 Page 1 000973 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-351 Page 165 Data File Acq On Sample Mise Quantitation Report IQT Reviewed] D:\MSD DATA\PAB\PABX-BBA\l-047-l\04710Bl .D Vial: 81 3 Jan 2001 14:29 Operator: 1103 6-GROUP11- FI Inst : MS7 MVL _______ _________________________Amountj______0.00 '"`ion': 131 (7~018> " ' ... " Resp: 260 0471081.D PABX BBA.M Thu Jan 18 15:11:47 2001 Page 2 009374 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-352 Page 166 Peak Area Report Sample Name: 11037-GROUP11-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 14:49 Operator Data File Name: 0471082.D Data File Path: D:\MSD_0ATAVPAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX.BBA Instrument Name: MS7 Instrument Vial Number: 82 Comoound # 1) 2) Comoound Name MVL MVL-4 Retention Time (Min.l 7.02 7.04 ton 131 135 Peek Area M* 187 1562 Peek Area Ratio 119.7E-3 M* - The letter 'r r r In this column Indicates that manual Integration eras performed on that compound. Print Date and Time: 1/18/01 15:11 Page 1 000975 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-353 Page 167 Data File Acq On Sample Mise Quantitation Report (QT Reviewed) D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471082.D Vial: 82 3 Jan 2001 14:49 Operator: 11037-GR0UP11-FI Inst : MS7 M V L __________ _________________________ Amount :_____ 0.0p ~~ Ion: 131 (7.018) Resp: 187 0471082.D PABX BBA.M Thu Jan 18 15:12:07 2001 Page 2 060 376 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-354 Page 168 Peak Area Report Sample Nam: 2 X Q C HIGH Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 15:09 Operator Data Rie Name: 0471011 .D Initrument Name: MS7 Data Rie Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument VkalNumber 11 Instrument Method Name: PABX_BBA Comnound # 1) 2) Comoound Name MVL MVL-04 Retention Time fMin.) 7.02 7.04 Ion 131 135 Peek Aree M* 1B74 1498 Peak Area Ratio 125.1E-2 M* - The letter "m"In this colum n indteatee that manual integration m i performed on that compound. Print Date and Time: 1/18/01 14:40 Page 1 000977 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-355 Page 169 Data F ile Acq On Sample Mise U iu an tieatian neputi. K evieoeu i D:\MSD DATA\PAB\PABX-BBA\l-047-l\0 4 7 1 0 1 1 .D V i a l : 11 3 Jan 2001 I S : 09 O perator: 2 X QC HIGH In st : MS7 0471011.D PABX BBA.M Thu Jan IB 14:4B:09 2001 Page 2 000978 Priniedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-356 Page 170 Peak Area Report Sample Name: Z X BLANK Notebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 15.29 O p e ra to r Data File Name: 0471016.0 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX_BBA Instrum ent Name: MS7 k is tru m e n t Vial N u m b e r 16 Compound * U 2) Com oound Name MVL M V l*d 4 R e te n tio n Tim e (Min.) 0.00 0.00 Ion 131 135 Peak Area M* 0 0 Peak Area Ratio #DtV/0! M' m The letter "m* In thte coktmn indicete* m et menue! integration was performed on that compound. Print Dale and Time: 1/1B/01 14:49 Page 1 000979 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-357 P age 171 Data F ile Acq On Sample Mise y u a n tita c io n Keporc ly r Kevxeweo; D:\MSD DATA\PAB\PABX - B B A \l-0 4 7 -l\0 4 7 10 16 .D V i a l : 16 3 Jan 2001 1 5 :2 9 O p erator: 2 X BLANK In st : MS7 MVL Amount : Not Found 0-471016.D PABX BBA.M Thu Jan 18 14:49:40 2001 Page 2 000980 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-358 Page 172 Peak Area Report Sample Name: 2 X CONTROL Notebook Reference: PABX-BBA-1-047 Date A cquited: 1/3/01 15:49 O perator Data R ie Name: 0471017.D Data R ie Path: D:\MSD_0ATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX.BBA Instrum ent Name: MS7 Instrum ent Vial N um b e r 17 Comoound# 1) 2) Compound Name MVL MVL-d4 Retention Time flltn.) 0.00 7.04 Ion 131 135 Peak Area M* 0 1533 Peak Anta Retto OOO.OE-3 M * The letter "m" In this colum n Indicete* that manual Integration was performed on that compound. Pnnl Dale and Time: 1/18/01 14:49 Page 1 000981 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-359 Page 173 Data F ile Acq On Sample Ml EC U u an titatio n xeport iut Kevieweai D:\MSD DATASPAB\PABX-BBA\1-0 4 7 - 1 \ 0 4 7 1 0 1 7 .D V i a l : 17 3 Jan 2001 15 :4 9 O perator: 2 X CONTROL Inst : MS7 M V L ______ ____ ______ ___ ; Ion: 131 (O'.OO) ' ' Amount j_Npt_ Found " ...................." 0471017.D PABX BBA.M Thu Jan 18 14:50:07 2001 Page 2 000982 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-360 Page 174 Peak Area Report Sample Name: 2 X STD 1 Notebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 16:09 O p e ra to r. Data FUe Name: 0471018.0 Data R ie Path: D:\MSD_DATA\PABtPABX-BBA\1-047-1\ Instrum ent Method Name: PABX_BBA Instrument Name: MS7 Instrum ent Vial N u m b e r 18 Comoound # 1) 2) Com oound Nome MVL MVL-d4 R eten tio n Time (Min.) 7.02 7.04 Ion 131 135 Peak Area M* 138 1688 Peek Area Ratio B17.5E-4 M* - The letter "m` in th*e colum n Indica ta that manual Integration wee performed on that compound. Print Data and Time: 1/18/01 14:50 Page 1 000983 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-361 Page 175 Q u an titatio n Report (UT Reviewed) Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471018 . D V i a l : 18 3 Jan 2001 2 X STD 1 16:09 O perator: Inst : MS7 I n t e g r a t i o n Pararne: RTEINT.P an t Method : D:\HPCHEM\l\METHODS\PABX_BBA.M IRTE I n t e g r a t o r) MVL _________ I o n :' 3 1 '(7.0 17) _______________________Amount : _ 0 ._00 0471018.D PABX BBA.M Thu Jan 10 14:50:27 2001 Page 2 000984 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-362 Page 176 Peak Area Report Sample Name: 2 X STD 2 N otebook Reference: PABX-BBA-1-047 Date A cquired: 1/3/01 16:29 Operator: Data File Name: 0471019.D Data File Path: D:\MSD_DATA\PAB\PABX-B8A\1-047-1\ instrum ent Method Name: PABX.BBA Instrum ent Name: MS7 Instrum ent Vial N u m b e r 19 Comoound # 1) 2) Comoound Name MVL MVL-d4 R eten tio n Time IMin.l 7.02 7.04 Ion \3\ 135 Peak Area M* 22B 1538 Peak Area Ratio 148.2E-3 M * The letter *m ' in this column indicates that manual integration was performed on that compound. Print Date and Time: 1/18/01 14:50 Page 1 000985 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-363 Page 177 Data F ile Acq On Sample Mise Quantitation Report 1QT Reviewed) D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471019.D V i a l : 19 3 Jan 2001 1 6 :2 9 O perator: 2 X STD 2 In st : MS7 MVL_______ __________ .___ ____ _____ Amount :______ 0.00 " ion: 13i (iToaoT i Resp: 220 0471019.D PABX BBA.M Thu Jan 18 14:50:47 2001 Page 2 oooyffG Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-364 Page 178 Peak Area Report Sample Name: 2 X STD 3 Notebook Reference: PABX-BBA-1-047 Date A cquired: 1/3AJ1 16:49 Operator: Data File Name: 0471020.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-11 ku tru m en t Method Name: PABX-BBA In itru m e n t Name: MS7 Instrum ent Vial Number: 20 Comoound # 1) 2) Comoound Name MVL MVL-cK R eten tio n Time (Min.) 7.02 7.04 Ion 131 135 Peak Area M* 386 1311 Peak Area Ratio 294.4E-3 M* s The letter "m Htn th is co lu m n m d icete e th a t m anual in tegration w aa p e rto rm e d o n th at com p o u n d . Print Date and Tm e: 1118/01 14:50 Page 1 000987 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-365 Page 179 Data File Acq On Sample Mise Uuantitation tteport nevieveai D:\MSDDATA\PAB\PABX-BBA\l-047-l\0471020.D Vial: 20 3 Jan 2001 16:49 Operator: 2 X STD 3 Inst : MS7 0471020.D PABX_BBA.M Thu Jan 18 14 :51:06 2001 Page 2 000988 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-366 Page 180 Peak Area Report Sample Name: 2 X STD 4 N otebook Reference: PABX-BBA-1-047 Date A cquired: 1/3/01 17:09 O p e ra to r Data File Name: 0471021.D Data File Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABX_BBA tnatrum ent Name: MS7 Instrum ent V ial N u m b e r 21 Comoound f i) 2) Com oound Nam MVL MVL-d4 R e te n tio n Tim e fMIn. 7.02 7.04 Ion 131 135 Peek Aree M* 565 1651 Peak Area Ratio 342.2E-3 M ' * The letter "m * In m is column Indicates that manual Integration m e pertonned on that com odate. Print Date and Time: 1/18/01 14:51 Page 1 000989 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-367 Page 181 Data File Acq On Sample Mise Quantitation Keport tur vieweai D:\MSD DATAIPAB\PABX-BBA\1-047-1\0471021.D Vial: 21 3 Jan 2001 17:09 Operator: 2 X STD 4 Inst : MS7 0471021.D PABX BBA.M Thu Jan 18 14:51:26 2001 Page 2 000990 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-368 Page 182 Peak Area Report Sample Name: 2 X STO S Notebook Reference: PABX-BBA-1-047 Date A cquired: 1/3/D1 1729 O perator Data File Name: 0471022.0 Data File Path: O:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX_BBA feislrum ent Name: MS7 In itn jm e n t Vial Num ber. 22 Comoound # i) 2) C om oound Name MVL MVL-d4 Retention Time fMin.l 7.02 7.04 ton 131 135 Peak Aree M' 936 1488 Peak Area Ratio 629.0E-3 M* * T h e letter 'm " tn th is c o lu m n In d lc e te e th a t m en u s! Integration erne pe rfo rm e d o n m at com pound. Print Dale and Time: 1/18/01 14:51 Page 1 000991 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-369 Page 183 Data File Acq On Sample Mise Quantication Report (QT Reviewed) D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471022.D Vial: 22 3 Jan 2001 17:29 Operator: 2 X STD 5 Inst : MS7 N V L _____ _________ Ion: 1377.09) Resp: 93 ______ .. . Amount : __ 0.00 0471022.D PABX BBA.M Thu Jan 18 14:51:47 2001 Page 2 000992 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-370 Page 184 Peak Area Report Sample Name: 2 X STD 6 N otebook Reference: PABX-BBA-1 -047 Date A cq u ire d : 1/3/01 17:48 O perator Data File Name: 0471023.0 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Inatrument Method Name: PABX_8BA Inatrum ent Name: MS7 Inatrument Vial N um b e r 23 Compound # U 2) Compound Name MVL MVL-d4 R eten tio n Tim e (Min.) 7.02 7.04 Ion 131 135 Peak Area M* 1415 1389 Peak Area Ratio 101.9E-2 M* * The tetter *m ' In th ie cak m tn In d icates th a t m an u al Integration w aa p artem iad o n th a t c o m p o u n d . Print Date and Time: 1/18/01 14:52 Page 1 000993 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-371 Page 185 Q u a n tita tio n Report IQT Reviewed) Data F ile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471023.D V i a l : 23 3 Jan 2001 1 7 :4 8 O perator: 2 X STD 6 Inst : MS7 i I n t e g r a t i o n Params: RTEINT.P huant Method : D:\HPCHEM\1\MKTK0DS\PABX_BBA.M (RTE I n t e g r a t o r ) M V L ____________ _______________________________ Amount :__________0.00 Io n : 13 1 H . 020)" 0471023.D PABX BBA.M Thu Jan 18 14:53:03 2001 Page 2 000994 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-372 Page 186 Peak Area Report Sample Name: 2 X STD 7 Notebook Reterance: PABX-BBA-1-047 Date A cquired: 1/3/01 18:08 O perator Data R ie Name: 0471024.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ Instrum ent Method Name: PABX_BBA kiatrum ent Name: MS7 Instrum ent Vial Num ber: 24 Comoound 4 1) Z) C om pound Name MVL MVL-d4 R e te n tio n Time /Min.) 7.02 7.04 Jen 131 135 Peak Area M* 2396 1444 Peak Area Ratio 165.96-2 M ' . The tetter *m* m this column indicates that manual kitagtetlan wee performed on that compound. Print Dale and Time: 1/18/01 14:53 Page 1 000995 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-373 Page 187 Data F ile Acq On Sample Mise y u a n t it a t io n Report (QT Reviewed) D:\MSD DATA\PAB\PAEX-BBA\l-047-l\0471024 -D V i a l : 24 3 Jan 2001 I S : 08 O p erator: 2 X STD 7 Inst : MS7 MVL _____________________________________________Amount :________0.00 Ion: 131 (7.017) : Resp: 2396 PABX BBA.M Thu Jan IB 1 4 :5 3 :2 3 2001 Page 2 000996 418-018:PAGE 1-374 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Study Title Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) in the Mevaionic Acid/Cholesterol Challenge and NOEL Investigation Study in Rats Analytical Laboratory Report Title Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) Fluorochemical in Rat Sera Samples Data Requirement Not Applicable Author 3M Environmental Laboratory Study Completion Date July 06, 2001 Performing Laboratories Sera Analyses 3M Environmental Laboratory Building 2-3E-09, 935 Bush Avenue St. Paul, MN 55106 Sera Extractions Pace Analytical Services, Inc.--Tier2 1700 Elm Street SE, Suite 100 Minneapolis. MN 55414 Project Identification 3M Medical Department Study: T-6295.25 Argus In-Life Study: 418-018 Analytical Report: FACT TOX-169 3M Laboratory Request No. E01-0180 Total Number of Pages 68 3M Environm ental Laboratory .'St Environmental Laboratory- 000997 Page 1 Page 1 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 418-018:PAGE 1-375 Analytical Report: F A C T -T O X -16 9 LR N -E01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 This page has been reserved for specific country requirements. 3 M Environm ental Laboratory 3\1 Environmental Laboraton- Page 2 000998 Page 2 418-018:PAGE 1-376 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 GLP Compliance Statement Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 Analytical Laboratory Report Title: Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) Fluorochemical in Rat Sera Samples Study Identification Numbers: T-6295.25, FACT TOX-169, LRN-E01 -0180 This study was conducted in compliance with United States Food and Drug Administration (FDA) Good Laboratory Practice (GLP) Regulations 21 CFR Part 58, with the exceptions in the bulleted list below. Exceptions to GLP compliance: There were two study directors for this study; one for the in-life phase and one for the analytical phase. The in-life study phase was considered to end at the generation and shipment of specimens. The analytical study phase was considered to start at the receipt of these specimens for analysis. Since the technical performance of each phase was entirely separate, no effect is expected from this exception. Automated data collection systems have not been validated as required according to 21 CFR 58.130(c). The hard copy printout is considered the raw data. The stability of the reference standard and internal standard were not determined. Deanna J. Luebker, M.S., Study Director 6L-t0 7 1 . ^ Marvin T. Case, D.V.M., Ph.D., Sponsor Representative l! u ln I Date Date j l Kristen J. Hansen, Ph.D., Principal Analytical Investigator JlcrJL C, 7<W I Date -- ' _______________________G 6 / 6 s~ / o / William K. Reagen, Ph.D., Laboratory Manager Date 3M Environm ental Laboratory 3M Environmental Laboratori' Page 3 000999 Page 3 418-018:PAGE 1-377 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 3M Medical Department Study: T-6295.25 Analytical Report: FACTTOX-169 LRN-E01-0180 GLP Study-- Quality Assurance Statement Analytical Laboratory Report Title: Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) Fluorochemical in Rat Sera Samples Study Identification Numbers: T-6295.25, FACT TOX-169, LRN-EOI -0180 This study has been inspected by the 3M Environmental Laboratory Quality Assurance Unit (QAU) as indicated in the following table. The findings were reported to the study director and laboratory management. Inspection Dates Phase Date Reported to Management Study Director 2/14/01 In-phase 2/16/01 2/16/01 2/28/01 In-phase 2/28/01 2/28/01 3/12/01-3/13/01 Data 3/14/01 3/14/01 4/20/01,4/23/01-4/24/01 Data 4/24/01 4/24/01 5/22/01-5/24/01 Draft report 5/24/01 5/24/01 QAU Representative -JI-OI Date 3M Environm ental Laboratory 3.1/ Environmental Laboratory Page 4 001000 Page 4 418-018:PAGE 1-378 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Table of Contents Analytical Report: FACT-TOX-169 LRN-EO1-01-80 Analytical Report: FACTTOX-169 LRN-E01-0180 GLP Compliance Statem ent..........................................................................................................3 GLP Study-- Quality Assurance Statem ent................................................................................4 Study Personnel and Contributors................................................................................................7 Introduction and Purpose.................................................................................................... ..........8 Test System ................................................................................................................................8 Specimen Collection and Analysis......................................................................................... 9 Specimen Receipt and Maintenance.......................................................................................... 9 Chemical Characterization of the Reference Substance........................................................ 10 Method Summaries................................................................................................................... . 11 3M Environmental Laboratory................................................................................................. 11 Preparatory Method............................................................................................................. 11 Analytical Method.................................................................................................................. 11 Analytical Equipment............................................................................................................ 12 Deviations................................................................................................................................... 13 Data Quality Objectives and Data Integrity................................................................................13 Data Summary, Analyses, and Results........................................................................................ 13 Summary of Quality Control Analyses Results.....................................................................14 Statement of Data Quality........................................................................................................14 Summary of Sample Results.................................................................................................... 15 Statistical Methods and Calculations........................................................................................... 15 Statement of Conclusion................................................................................................................15 Appendix A: Control Matrices and Test Article.......................................................................... 16 Appendix B: Protocol and Deviations........................................................... .............................. 17 Appendix C: Extraction and Analytical Methods.........................................................................3 Q ETS-8-4, Extraction of Fluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry, (15 pages)..................................................................31 ETS-8-5, Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry, (11 pages)........................ 4 6 Appendix D: Data Summary Tables.............................................................................................. 5 7 Appendix E: Data Spreadsheets.................................................................................................... 5 5 Appendix F: Example Calculations.............................................................................................. 63 Appendix G: Interim Certificate of Analysis............................................................................... 64 Appendix H: Report Signature Page........................................................................................... 6 8 3 M Environm ental Laboratory 5M Environmental Laboratory Page 5 O O iO O i Page 5 418-018:PAGE 1-379 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 List of Tables Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Table 1. Female Rat FO Population Demographics for Study (Argus 4 1 8 -0 1 8 )............... 8 Table 2. Characterization of the Analytical Reference Substance in Study FACT TOX169........................................................................................................................................ 10 Table 3. Target Ions Monitored in 3M Laboratory Analyses.................................................... 12 Table 4. Determinations of the LOQ in the Analyses of PFOS Extracts............................... 14 Table 5. Characterization of the Control Matrices Used for Sera Analyses in Study FACT T O X -169...................................................................................................................16 Table 6. Characterization of the Test Article in Study FACT T O X -1 6 9 ..................................16 Table 7. TOX 169 Data Summary of PFOS Concentration-- Rat Serum (pg/mL).............. 5 7 3 M Environm ental Laboratory 3 \l Environmental Laboratori- Page 6 001002 Page 6 418-018:PAGE 1-380 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 3M Medical Department Study: T-6295.25 Analytical Report: FACT TOX-169 _______________________________________________________________ LRN-E01-0180 Study Personnel and Contributors Study Director Deanna J. Luebker, M.S. 3M Corporate Toxicology (Medical Department) Building 220-2E-02 St. Paul, MN, 55144 Analytical Chemistry Laboratories S era Analyses 3M Environmental Laboratory (3M Lab) Kristen J. Hansen, Ph.D., P rincipal A nalytical Investigator Sponsor 3M Corporate Toxicology 3M Medical Department Building 220-2E-02 St. Paul, MN, 55144 Marvin T. Case, D.V.M., Ph.D., Sponsor Representative S era Extractions Pace Analytical Serv ces, Inc. Tier2 Facility 3M Lab Contributing Personnel Lisa A. Clemen Rhonda Dick* Marlene M. Heying* Harold O. Johnson Kelly J. (Kuehlwein) Swartout* Bob W . Wynne* 'Contract lab professional service employees Location of Archives All original raw data, protocol, and analytical report have been archived at the 3M Environmental Laboratory. The test article and analytical reference standard reserve samples, as well as the specimens pertaining to the analytical phase of this study are archived at the 3M Environmental Laboratory. Reserve samples of the control article and vehicle components are stored at the testing facility (Argus). 3 M Environm ental Laboratory 3M Environmental Laboratory- Page 7 001003 Page 7 418-018:PAGE 1-381 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 Introduction and Purpose The purpose of the study is to determine the presence and concentration of perfluorooctanesulfonate (PFOS) in rat sera samples taken in accordance with the 3M protocol, "Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) in the Mevalonic Acid/Cholesterol Challenge and NOEL Investigation Study in Rats." Female FO generation rats were given PFOS by oral gavage and maternal and offspring serum samples were collected for analysis at specified intervals. The intent of the study was to determine the maternal and fetal effect of the co-administration of mevalonic acid or cholesterol given with PFOS doses, as well as to better define the PFOS `no observed effect level' (NOEL). This study was initiated 31 January 2001. T e st System Thirteen groups of female rats were used as the test system. Group 1, Group 2 and Group 5 were control groups that were given either Tween 80, mevalonic acid, or cholesterol only. Groups 3, 4, and 6 -- 13 were administered PFOS doses in 0.5% Tween 80 (test article), either alone or in combination with mevalonic acid or cholesterol. Female rats were given the test article beginning 42 days prior to cohabitation, and continuing through day 20 of presumed gestation (Caesarean section group), day 24 of presumed gestation (rats assigned to natural delivery that do not deliver a litter) or day 4 postpartum (rats that deliver a litter). The test system species and strain selected was the Crl:CD*(SD) IGS BR VAF/Plus rat received from Charles River Laboratories, Inc., permanently identified using a Monel self-piercing ear tag. Only F0 generation rats were identified with ear tags. F1 generation rats were identified and evaluated as part of litters, but were not individually tagged, and all parameters were evaluated in terms of the litter (see Table 1). Table 1. Female Rat F0 Population Demographics for Study (Argus 418-018) Dosage Groups Population Selected for Study Dosage Vehicle Control Group 1 Mevalonic Acid Control G roup 2 Group 3 Group 4 Cholesterol Control Group 5 Group 6 Group 7 G roupfl Group 9 Group 10 G roup 11 Group 12 Group 13 2B 28 28 28 28 28 28 20 20 20 20 28 28 0.5% Tween 80 mevalonic acid 1.6 mg/kg PFO S + mevalonic acid 2 mg/kg PFOS + mevalonic acid cholesterol 1.6 mg/kg PFO S + cholesterol 2 mg/kg PFO S + cholesterol 0.4 mg/kg PFOS 0.8 mg/kg PFO S 1 mg/kg PFOS 1.2 mg/kg PFO S 1.6 mg/kg PFO S 2 mg/kg PFOS 3 M Environm ental Laboratory 3.\t Environmental Laboratori Page 8 001004 Page 8 3M Medical Department Study. T-6295.25 3M Medical Department Study: T-6295.25 418-018:PAGE 1-382 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 Specim en C ollection and Analysis In the analytical study reported here, 235 sera specimens collected from female FO rats and F1 pooled rat litters were sent to the 3M Environmental Laboratory and analyzed for PFOS. The number of sera specimens collected for analyses in the analytical phase of this study are presented below. Specimens Collected from Study Groups 1 through 13: Sera Specim ens-- 54 specimens (FO Females, G D21, Caesarean section) Sera Specim ens-- 54 specimens (F1 pooled litter, G D21, Caesarean section) Sera Specim ens-- 78 specimens (FO females, day 5 of lactation, natural delivery) Sera Specimens-- 49 specimens (F1 pooled litter, day 5 of lactation, natural delivery) Blood specimens were centrifuged. Serum was then harvested and shipped to the 3M Environmental Laboratory frozen and on dry ice. Sera samples were extracted beginning on 05 February 2001 using an ion pairing reagent and methyl-ferf-butyl ether (MtBE). Sample extracts were analyzed using high-pressure liquid chromatography-electrospray/tandem mass spectrometry (HPLC-ESMSMS) in the multiple response monitoring mode. PFOS levels were quantitated by external calibration versus an extracted curve. Analytical details are included in this report. Specimen Receipt and Maintenance The 3M Environmental Laboratory received rat serum specimens collected at Argus Research Labs. All specimens were received frozen in good condition on dry ice and were immediately transferred to storage at -20C 10C. Specimens were transported to Pace Tier2 frozen on ice packs. After extraction at Tier2, the specimens and the extracts were returned cold on ice packs. Control matrices used in sera analyses performed during TOX-169 were obtained from commercial sources and are presented in Appendix A (see Table 5). Samples analyzed at the 3M Environmental Laboratory will be maintained for a period of 10 years and will be stored at the laboratory at -20C 10C. 3M EnvironmentalLaboratory 3M Environmental Laboraron Page 9 001005 Page 9 3M Medical Department Study: T-6295.25 3M Medical Department Study. T-6295.25 418-018:P AGE 1-383 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Chemical Characterization of the Reference Standard Potassium Peril uorooctanesulfonate (KPFOS) CAS Number: 2795-39-3 Chemical Formula: CgF^SC^'K* Molecular Weight: 537.9 Chemical characterization information on the reference substance potassium perfluorooctanesulfonate used in this study is presented in tabular form below. Table 2. Characterization of the Analytical Reference Substance in Study FACTTOX-169 R eference S u b stan ces Chemical Name KPFOS (Potassium Perfluorooctanesulfonate)* Source 3M C orporate Toxicology. Medical D epartm ent E xpiration D ate 8/31/01 Storage Conditions Chemical Lot Number Frozen <-10C 217 Physical Description W hite crystalline powder Purity 86.9% Target analyte is PFOS, C.FuSOs' TBO--To be determined THPFOS (Tetrahydrooerfluorooctanesulfonic acid) TCFI00017-055 SynOuest Labs 2005 -20C Q75-91 W hite powder TBD* 3M EnvironmentalLaboratory .'M Environmental Laboratory Page 10 001006 Page 10 418-018:PAGE 1-384 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Method Summaries Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 Following is a brief description of the methods used during this analytical study by the 3M Environmental Laboratory. Detailed descriptions of the methods used in this study are located in Appendix C. 3M Environm ental Laboratory PreparatoryMethod ETS-8-4.2, "Extraction of Fluorochemical Compounds from Serum for Analysis using HPLCElectrospray/Mass Spectrometry" Analytical serum samples were extracted using an ion-pairing extraction procedure. An ion pairing reagent was added to a laboratory sample and the analyte-ion pair was partitioned into MtBE. The MtBE extract was then removed and put onto a nitrogen evaporator until dry. Each extracted laboratory sample was reconstituted in 1.0 mL of methanol and filtered through a 3 mL plastic syringe attached to a 0.2 pm nylon filter into glass autovials. AnalyticalMethod ETS-8-5.2, "Analysis of Potassium Perfluorooctane-sulfonate or other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" The analyses were performed by monitoring one product ion selected from a single primary ion characteristic of a particular fluorochemical using HPLC-ESMSMS. For example, molecular ion 499, selected as the primary ion for PFOS (CBFi7S03-) analysis, was fragmented to produce ion 99 (F S 0 3-). The characteristic ion 99 was monitored for quantitative analysis. 3M Environmental Laboratory 3\1 Environmental Laboraron' Page 11 001007 Page 11 418-018:PAGE 1-385 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 AnalyticalEquipment The following is representative of the settings used during the analytical phase of this study. Liquid Chrom atograph: Hewlett-Packard Series 1100 Liquid Chromatograph system Analytical column: Keystone BetasilTM C,( 2x50 mm (5 pm) Column temperature: Ambient Mobile phase components: Component A: 2mM ammonium acetate Component B: methanol Flow rate: 300 pL/min Injection volume: 10 pL Solvent Gradient: 10.0 minutes (minutes) 0.0 1.0 5.5 7.5 8.0 %B 10% 10% 95% 95% 10% Mass Spectrometer: Micromass API/Mass Spectrometer Quattro IITM Triple Quadrupole system Software: Mass LynxTM 3.4 Cone Voltage: 60 V Collision Gas Energy: 4 0 -4 5 eV Mode: Electrospray Negative Source Block Temperature: 150C 10C Electrode: 2-spray Analysis Type: Multiple Reaction Monitoring (MRM) Table 3. Target Ions Monitored in 3M Laboratory Analyses T a rg e t A n alyte P rim a ry Ion (a m u ) P ro d u c t Io n (a m u ) PFOS THPFOS 499.0 427.0 99.0 BO.O 3M EnvironmentalLaboratory 3M Environmental Laboratory Page 12 001008 Pa& 12 418-018:PAGE 1-386 3M Medical Department Study: T-6295.25 3M M edical D ep artm en t Study: T -6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 A nalytical R eport: FACT TO X -169 L R N -E Q 1-0180 D e v ia tio n s Deviations from the original protocol and methods are included in the Appendix B. Data Quality Objectives and Data Integrity The following data quality objectives (DQOs) were indicated in the protocol for this study: Linearity: The coefficient of determination (rz) equal to or greater than 0.990 using 1/x weighting. Limits of Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the calibration curve, defined as the lowest standard that is both two times the matrix blank and is calculated within 30% of the expected concentration. Acceptable Precision: Quality control samples are required to meet 25% precision standards, provided they are quantitated within the selected calibration range. Confirmatory Methods: If a confirmatory method is used, an amendment to this protocol should be written. Demonstration of Specificity: PFOS identification will be substantiated by chromatographic retention time (approximately 7.3 minutes), by the characteristic primary ion (499) and the characteristic product ion (99). Data Summary, Analyses, and Results Data quality objectives for the analytical phase of this study outlined in the 3M Environmental Laboratory protocol for FACT TOX-169 (see Appendix B) were met with the exceptions noted in this report. 3M Environmental Laboratory 331 Environmental Laboraton.' Page 13 001009 Page 13 418-018:PAGE 1-387 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 Summary of Quality Control Analyses Results Linearity: The coefficient of determination (r2) of the standard curve was 0.990. Calibration Standards: Quantitation of the target analytes was based on linear regression analysis (1/x weighted) of an extracted rat sera matrix curve run prior to each group of samples. High or low points on the curve may have been deactivated to provide a better linear fit over the curve range most appropriate to the data. Low curve points with peak areas less than two times that of the extraction blanks were deactivated to disqualify a data range that may have been significantly affected by background levels of the analyte. Occasionally, a single mid-range curve point that was an obvious outlier may have been deactivated. Quantitation of each analyte was based on the response of one specific product ion using the multiple response-monitoring mode of the instrument (see Appendix C, Analytical Methods). Limits of Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the calibration curve (defined as the lowest valid standard within 30% of the theoretical value), and is at least two times the analyte peak area detected in the matrix blanks. (See below.) Table 4. Determinations of the LOQ in the Analyses of PFOS Extracts Analyte Method LOQ PFOS 0.00494 pg /mL Blanks: All blanks were below the lower limit of quantitation for the compounds of interest. Precision: Precision was determined by analysis of extracted calibration check standards, which were reproducible to within 25% over the valid linear range. Instrument Precision: Instrumental precision was determined by replicate (n=7) injections of a single serum extract. Peak area was reproducible to within 2%. Internal Standards: The internal standard (THPFOS) was added to all samples and standards. THPFOS was not used for quantitation, but was used to monitor for gross instrument failure. The surrogate response of each analytical run was verified to determine that it did not vary more than 50% from the mean within each analytical run. Samples with deviant internal standard response were confirmed by reanalysis and are marked in the table of results. Statement of Data Quality It is not possible to verify true recovery of endogenous analyte from tissues without radio-labeled reference material. The only measurement of accuracy available at this time, extracted calibration curves and multi-level continuing calibration verifications (CCVs), indicate that the sera data are quantitative to 25% or greater. 3M EnvironmentalLaboratory 5 \t Environmental Laboratori' Page 14 001010 Page 14 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 418-018:PAGE 1-388 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Summary of Sample Results PFOS results have been corrected for purity of the analytical reference material. Samples from Control Animals: Low levels of PFOS were often detected in the sera of the control animals. These levels were significantly lower than those found in the low dose test animals. Samples from Dosed Animals: In general, PFOS levels found in the sera of the test animals increased with dose level. Detailed sample data tables are presented in Appendices D and E. Statistical Methods and Calculations Statistical methods were limited to the calculation of means and standard deviations. See Appendix F for example calculations used to generate the serum sample data in FACT TOX169. Statement of Conclusion Under the conditions of the present studies, the fluorochemical PFOS was observed in the sera of all FO generation rats dosed with the test article and in all F1 generation rats from FO dosed rats. 3M EnvironmentalLaboratory 3M Environmental Laboratori' Page 15 OOlOii Page 15 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 418-018:PAGE 1-389 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Appendix A: Control Matrices and Test Article Table 5. Characterization of the Control Matrices Used for Sera Analyses in Study FACT TOX-169 Control Matrix Source TCR # Expiration Date Storage Conditions Chemical Lot # Physical Description Rat Serum Sigma Chemicals 99062-9 01/01/2010 Frozen -20C 17H9306 Rat Serum Rat Serum Sigma Chemicals 99062-31 01/01/2010 Frozen -20C 17H9306 Ral Serum Rat Serum Sigma Chemiitals 99131-9 01/01/2010 Frozen -20C 19H89291 Rat Serum Rat Serum Sigma Chemicals 99131-11 01/01/2010 Frozen -20C 19H89291 Rat Serum Table 6. Characterization of the Test Article in Study FACT TOX-169 Location 3M Laboratory Test Article Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity Potassium Perfluorooctanesulfonate 3MCorporateToxicology, Medical Department 8/31/01 FrozenS-10C 217 White crystallinepowder 86.9% 3M Environmental Laboratory 5 \t Environmental Laboratory' Page 16 001012 Page 16 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Appendix B: Protocol and Deviations 418-018-.PAGE 1-390 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 3M EnvironmentalLaboratory 3 \ t Environmental Laboratory Page 17 001013 Page 17 3M Medical Department Study: T-6295.25 3M Environmental Technology and Services PO Box 33331 St. Paul. MN 55133-3331 612 778 6442 418-018:PAGE 1-391 Analytical Report: FACT-TOX-169 3M Phase Title Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) in the Mevalonic Acid/Cholesterol Challenge and NOEL Investigation Study in Rats PHASE PROTOCOL Perform ing Laboratories 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Pace Analytical Services, Inc. 1700 Elm Street SE, Suite 100 Minneapolis, MN 55414 Laboratory Project Identification FACT-TOX Number TOX-169 3M Toxicology Study Number T6295.25 Argus In-life Study Number 418-018 3M EnvironmentalLaboratory M Environmental Laboratori Page 1of9 061014 Page 18 3M Medical Department Study: T-6295.25 418-018.PAGE 1-392 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol FACT-TOX-169 Study Identification Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) in the Mevalonic Acid/Cholesterol Challenge and NOEL Investigation Study in Rats Sponsor 3M Corporate Toxicology 3M Medical Department Building 220-2E-02 St. Paul, MN 55144 Sponsor Representative Marvin T. Case D.V.M., Ph.D. 3M Corporate Toxicology 3M Medical Department Building 220-2E-02 St. Paul, MN 55144 Telephone: (651)733-5180 Study D irector Phase Locations Principal Analytical Investigator (PAI) Deanna J. Luebker M S. 3M Corporate Toxicology 3M Medical Department Building 220-2E-02 St. Paul, MN 55144 Telephone: (651)737-1374 Kristen J. Hansen, Ph.D. Analytical Testing Laboratories 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN55106 Proposed Phase Tim etable Analyses Start Date Analyses Termination Date Pace Analytical Services, Inc. 1700 Elm Street SE, Suite 100 Minneapolis, MN 55414 January 20,2001 February 28,2001 3M EnvironmentalLaboratory i.U E n v ir o n m e n ta l L a b o r a to r y Page 2 of9 001015 Page 19 3M Medical Department Study: T-6295.25 418-018:PAGE 1-393 A nalytical Report: FA C T -T O X -169 L R N -E 01-0180 Protocol F A C T -T O X -1 69 1. Study Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) in the Mevalonic Acid/Cholesterol Challenge and NOEL Investigation Study in Rats 2. Purpose The analytical phase of this study is designed to evaluate levels of PFOS in sera specimens of FOgeneration female rats exposed and FI generation rat pups potentially exposed to PFOS in Argus In-life Study 418-018. All sera samples will be extracted at Pace Analytical Services Inc. Tier II facility. All sera samples will be analyzed at the 3M Environmental Laboratory. Data quality objectives for this study are indicated in section 12. 3. Regulatory Compliance This study will be conducted in accordance the United States Food and Drug Administration Good Laboratory Practice Regulations for Non-clinical Laboratory Studies, Final Rule 21 CFR Part 58. 4. Quality A ssurance The 3M Environmental Laboratory Quality Assurance Unit will audit the study conduct, raw data, and final report to determine compliance with Good Laboratory Practice Regulations, this protocol, and 3M Environmental Laboratory Standard Operating Procedures. 5. Reference Standard For FACT-TOX-169 Potassium Perfluorooctanesulfonate (KPFOS), C8F17SO3K, CAS 2795-39-3 6. Test System Thirteen dose groups o f female rats were given the test article by oral (gavage) route. Within each group, one blood sample was collected from six female FOon day 21 of gestation (at Caesarean sectioning), from six female FOon day 5 of lactation (after natural delivery), from six FI pooled litters on day 21 of gestation (at Caesarean sectioning), and from six FI pooled litters from day 5 of lactation (after natural delivery). There are 312 sera specimens sent to the 3M Environmental Laboratory. See following table for a description of all samples received. 3M Environmental Laboratory 3M Environmental Laboratory Page 3 of 9 001016 Page 20 3M Medical Department Study: T-6295.25 418-018:PAGE 1-394 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol F A C T -T O X -169 Number of Specimens per Dosage group and Dosage Levels D osage Group and Level 1 Vehicle Control 2 M evalonic Acid C o n tro l 3 1.6 m g/kg PFOS + M evalonic Acid 4 2.0 m g/kg PFOS + M evalonic Acid 5 Cholesterol C o n tro l 6 1.6 m g/kg PFOS C holesterol T 2.0 m g/kg PFOS + Cholesterol B 0.4 m g/kg PFOS 9 0.8 m g/kg PFOS 10 1.0 m g/kg PFOS 11 1.2 m g /k g PFO S 12 1.6 m g/kg PFOS 13 2.0 m g/kg PFOS F0 R ats from C aesarean S e c tio n Day 21 6 6 6 6 6 6 6 6 6 6 6 6 6 F1 R at P u p s from C aesarean Section Day 21 6 6 6 6 6 6 6 6 6 6 6 6 6 F0 Rats from Lactation Day 5 6 6 6 6 6 6 6 6 6 6 6 6 6 F1 R at P u p s from Lactation Day 5 6 6 6 6 6 6 6 6 6 6 6 6 6 7. Specimen Receipt Sera specimens will be received from Argus Research Laboratories, Inc. at the end of the in life phase on January 2001 (estimated). Sera; specimens were shipped by overnight courier and were received frozen on dry ice. Specimens were identified with sample numbers (lab request numbers or numbers assigned by Argus Research Laboratories, Inc.) upon receipt, registered with the 3M Environmental Laboratory and transferred to a freezer for storage. All specimens sent to the 3M Environmental Laboratory were received and tracked according to the Sample Tracking System Standard Operating Procedure. Detail.1of specimen inspection for damage, receipt, storage, identification, chain of custody protocols, and data will be presented in the phase report. 8. Preparatory Methods ETS-8-4, "Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemicals Compounds from Serum for analysis using HPLC-Electrospray/Mass Spectrometry" Analytical samples are extracted using an ion-pairing extraction procedure. An ion-pairing reagent is added to a laboratory sample and the analyte ion pair is partitioned into MtBE. The MtBE extract is then removed and put onto a nitrogen evaporator until dry. Each extracted laboratory sample is reconstituted in 1.0 mL of methanol and filtered through a 3 cm3 plastic syringe attached to a 0 .2 pm nylon filter into glass autovials. 3M Environmental Laboratory 3M Environmental Laboratory Page 4 of 9 001017 Page 21 418-018:PAGE 1-395 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol F A C T -T O X -169 9. Analytical Methods ETS-8-5, "Analysis ofPotassium Perfluorooctanesulfonate or other Fluorochemicals in Serum Extracts using HPLC-Electrospray/Mass Spectrometry" Analysis is performed by monitoring one or more product ions selected from a single primary ion characteristic of a particular fluorochemical using HPLC-ESMSMS. For example, molecular ion 499, selected as the primary ion for PFOS (CgFnSC>3-) analysis, is fragmented further to produce ion 99 (FS03-). The characteristic ion 99 is monitored for quantitative analysis. Minor modifications to the preparatory and analytical methods may be implemented at the discretion of the PAL 10. Data Quality Objectives 10.1 Linearity The coefficient of determination (r2) of the standard curve must be equal to or greater than 0.990 using 1/x weighting. 10.2 Lim its o f Q uantitation (LOQ) The LOQ will be equal to the lowest acceptable standard in the calibration curve, defined as the lowest standard that is both 2 times the matrix blank and is calculated within 30% of the expected concentration. 10.3 A c c e p ta b le P re c is io n Quality control samples are required to meet 25% precision, provided they are quantitated within the selected calibration range. 10.4 U s e o f C o n firm a to ry M e th o d s If a confirmatory method is used, an amendment to this protocol will be written. 10.5 D e m o n s tra tio n o f S p e c ific ity , etc. PFOS identification will be substantiated by chromatographic retention time (approximately 7.3 minutes), by the characteristic primary ion (499) and the characteristic product ion (99). Minor modifications to the Data Quality Objectives may be implemented at the discretion ofthe PAI. 3M Environmental Laboratory 3M Environmental Laboratory Page 5 of 9 001018 Page 22 418-018:PAGE 1-396 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol F A C T -T O X -169 11. Sub-Contracted Analysis Pace Analytical Services, Inc. (Tier II Facility) will be performing the extraction of serum specimens for analyses. All extracted samples will be shipped to the 3M Environmental Laboratory for analysis. Pace Analytical Services, Inc. will follow all applicable 3M Environmental Laboratory Standard Operating Procedures and the following Pace Analytical Inc. Standard Operating Procedures. PSS-Admin-01 Quality System PSS-DC-05 Lab Notebooks, Logbooks, and Phone Logs PSS-OPS-Ol Use and Maintenance of Ultra-Turrax T25 Homogenizer PSS-OPS-02 Raw Data PSS-OPS-03 Hazardous Waste Disposal PSS-OPS-04 Use and Maintenance of N-Evap Analytical Evaporator PSS-OPS-05 Use of Compressed Gases in the Laboratory PSS-OPS-06 Use and Maintenance ofNANOpure II Water Purification System PSS-OPS-07 Cleaning Non-disposable Volumetric Pipettes PSS-OPS-08 Laboratory Calculations PSS-OPS-13 Use and Maintenance of Lab Refrigerators, Freezers, and Ovens PSS-OPS-15 Glassware Cleaning PSS-OPS-16 Use and Maintenance of the Fisher Scientific Isotemp Freezer PSS-OPS-31 Use and Maintenance of Shakers PSS-OPS-32 Operation, Maintenance and Calibration of Laboratory ETS-9-40 Operation and Maintenance of Shakers and Vortex Mixers (3M SOP) PSS-OPS-37 Operation, Maintenance, and Calibration of pH Meters and pH Electrodes PSS-MTR-01 Calibration of Certified Weight Set PSS-MTR-02 Calibration of Certified Thermometers/Thermocouples PSS-MTR-06 Verification of Calibration ofthe Eppendorf Pipettor PSS-MTR-08 Verification of Calibration and Use of Analytical Balances PSS-MC-01 Purchasing of Laboratory Supplies PSS-ARC-02 Disposition of Archive Materials PSS-DM-03 Statistical Evaluation of Data 12. Statistical Analysis Statistical methods will be limited to the calculation of means and standard deviations. Examples ofthe calculations used in the analyses will be included in the phase report. 13. Report A report of the analyte results will be prepared by 3M Environmental Laboratory. The report will include, but not be limited to, the following, when applicable: 13.1 Name and address of the facility performing the phase. 13.2 Dates upon which the phase was initiated and completed. 13.3 A statement of compliance by the PAI addressing any exceptions to Good Laboratory Practice Standards. 3M Environmental Laboratory 3M Environm ental L aboratory Page 6 of 9 001019 Page 23 3M Medical Department Study: T-6295.25 418-018:PAGE 1-397 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol F A C T -T O X -169 13.4 Objectives and procedures as stated in the approved phase protocol, including any amendments to the original phase protocol. 13.5 Identity, purity, stability and the solubility of the reference standard under the conditions of administration. 13.6 A description of the methods used to conduct the test(s). 13.7 A description of the specimens. 13.8 A description of any circumstances that may have affected the quality or the integrity of the data. 13.9 The name of the PAI and the names of other scientists, professionals, and supervisory personnel involved in the phase. 13.10 A description of the transformations, calculations, or operations performed on the data, a summary and analysis of the analytical chemistry data, and a statement of the conclusions drawn from the analyses. 13.11 Statistical methods used to evaluate the data, if applicable. 13.12 The signed and dated reports of each of the individual scientists or other professionals involved in the phase, if applicable. 13.13 The location where raw data and the phase report are to be stored. 13.14 A statement prepared by the Quality Assurance Unit listing the dates that study inspections and audits were made and the dates of any findings reported to the Study Director and Management. 13.15 If it is necessary to make corrections or additions to a final report after it has been accepted, the changes will be made in the form of an amendment issued by the Study Director. The amendment will clearly identify the part of the final report that is being amended, the reasons for the amendment, and will be signed by the Study Director. 14. Location of Raw Data, Records, and Final Report Original data or copies thereof, will be available at 3M Environmental Laboratory to facilitate audits of the study during its progress and before acceptance of the phase report. When the phase report is completed, all original paper data, including those items listed below will be retained in the archives of 3M Environmental Laboratory. All corresponding training records, calibration records, instrument maintenance logs, standard operating procedures, equipment procedures, and methods will be retained in the archives of the 3M Environmental Laboratory. 14.1 The following raw data and records will be retained in the study folder in the archives according to 3M Environmental Laboratory Standard Operating Procedures. 14.1.1 Approved protocol and amendments 14.1.2 Study correspondence 14.1.3 Shipping records 14.1.4 Raw data 14.1.5 Approved final report (original signed copy) 14.1.6 Electronic copies of data 14.2 The following supporting records will be!retained separately from the study folder in the archives according to 3M Environmental Laboratory Standard Operating Procedures: 3M Environmental Laboratory 3M E nvironm ental L aboratory Page 7 of 9 001020 Page 24 3M Medical Department Study: T-6295.25 418-018.-PAGE 1-398 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol FACT-TOX-169 14.2.1 Training records 14.2.2 Calibration records 14.2.3 Instrument maintenance logs 14.2.4 Standard Operating Procedures, Equipment Procedures, and Methods 15. Data/SampleRetention Ail sera specimens remaining after the analytical phase is completed will be sent to and maintained by: Dave Ehresman 3M Corporate Toxicology 3M Center Bldg 236-0C-148 SL Paul, MN 55144 Telephone (651) 733-5070 Fax (651) 737-4754 16. ProtocolAmendmentsanddeviations Planned changes to the protocol will be in the form of written amendments signed by the Study Director and the Sponsor's Representative. Amendments will be considered as part of the protocol and will be attached to the final protocol. All changes to the protocol will be indicated in the phase report. Any other changes will be in the form of written deviations, signed by the Study Director and filed with the raw data. 17. A ttachments 17.1 A ttac h m e n t A : Material Safety Data Sheets (MSDS) for Reference Standard 3M Environmental Laboratory SM Environmental Laboratory Page 8 of 9 001021 Page 25 3M Medical Department Study: T-6295.25 18. Signatures 418-018:PAGE 1-399 Analytical Report: FACT-TOX-169 Protocol F A C T - T d < m E 0 1- 180 Y y i, ^ t & v - _______________ M arvin T Case, D . V.M., Ph.D ., S p on sor R epresentative j/% . ` Date 1 D eanna J. Luebker^M .S., Study D irector l (slLq D ate -- -- ~ ~ ______________________ K risten J. Hansen, Ph.D ,, Principal A nalytical Investigator 01 / z o lfa i D ate 3 M Environm ental Laboratory 5.U E n v ir o n m e n ta l L a b o r a to r \' Page 9 of 9 001022 Page 26 418-018:PAGE 1-400 3M Medical Department Study: T-6295.25 Record of Deviation A nalytical Report: FA C T -T O X -169 L R N -E 01-0180 t. Id e n tific a tio n Study / Project No. FA C T-TO X -169, Lim s # E 01-0180, Argus 418-018 D eviation Type Gsop Q M ethod Q Equipment Procedure (Check one) X P rotocol Q Other: Document Number FACT-TO X-169 Date(s) o f occurrence 02/01/01 II. D escription: Required Procedure/process: T he protocol states that 312 specim ens w ill be received at the Environm ental Laboratory. Actual Procedure/process: 235 specim ens w ere received at the Environm ental Laboratory. III. A ctions Taken: (such as amendment issued, SOP revision, etc.) This deviation w as written. Recorded B y D ate IV. Im pact on S tudy / P roject N o adverse im pact on the outcom e o f this study. 01 )L 05-/2H [0 Authorized B y (Study D irector /P ro je c t Lead) XJ%. C*- AWta. * <l Date . s /^ /o / D<itc.hsr Desnrta, Lueger Mw/i* Cwt. 3M Environmental Laboratory Form ETS-4-8.0 3)1*2,?+*/ Deviation No. (assigned by Study Director or Projott Lead at the end o f study or project) 3M E n vironm en tal Laboratory' 001023 Pas * 27 418-018:PAGE 1-401 3M Medical Department Study: T-6295.25 Record of Deviation Analytical Report: FACT-TOX-169 LRN-E01-0180 I. Id e n tifica tio n Study / Project No. FACT-TOX-169, Lims # EQ1-0180, Argus 418-018__________ ____ Deviation Type SOP ~jMethod Equipment Procedure (Check one) Protocol Other: Document Number ETS-8-4.2 Date(s) of occurrence 2/15/01 -2/21/01 II. D e s crip tio n : Required Procedure/process: 1) Section 10.1.2 and 10.1.3.2 Prepare two water blanks and two matrix blanks for each study. 2) Section 11.1.4 Prepare and spike one standard curve for each study. 3) Section 1 1.1.2 Do not extract less than Q.5mL of matrix. 4) Section 11.1.2 If most sample volumes are less than 1 .0 mL, extract standards with matrix volumes equal to the sample volumes. __ ______ ___ ______ __ Actual Procedure/process: 1) Six water blanks and six method blanks were added due to the large number of samples. 2) Four extracted curves were prepared - three with initial and final volumes of 0.5 mL and one with initial and final volumes of 1.0 mL. 3) Many samples were extracted with an initial volume less than 0.5 mL. Refer to the attached Sample volume/weight sheet for initial volume information.______ ______ _____ 4) For most samples, <0.5 mL of sera was available. However, curves were not prepared with an initial or final volume under 0.5 mL. III. A ctio n s Taken: (such as amendment issued, SO P revision, etc.) This deviation was written. Recorded By Date Kelly Swartout % A jrx ^ - n iii's IV. Im p act on S tudy / P roject 02/21/01 a ja a lo ) 1 & 2) Additional QC is an improvement and will not adversely affect this study. 3) Because the method is not characterized for volumes <0.5 mL, samples with initial volumes less than 0.5 mL will be flagged in the data. 4) This course of action was taken because the method has not been characterized for extractions <0.5mL. Robustness of the curves will not be adversely afflicted. ___ fc h OX/CL^lQi Authorized By (Study Director / Project Lead) Date 3M Environmental Laboratory Form ETS-4-3.0 OJlJov Deviation No. (assigned by Study Director or Project Lead at the end o f study or project) Q.ftc/tor Krr#- 0e.aiv\<y. L u t t e r 3M E nvironm ental Laboratory 001024 Page 28 418-018:PAGE 1-402 3M Medical Department Study: T-6295.25 Record of Deviation Analytical Report: FACT-TOX-169 LRN-E01-0180 Study / Project No. Deviation Type (Check one) Document Number /. Id e n tific a tio n SOP "ETMethod Equipment Procedure Protocol DVOther: Date(s) of occurrence II. D escription: Required Procedure/process: / } ( &yrzz*nn( ^ M S - 7 oe - 2 L z z ^ 2 - D J . -- = f e - <y f c c fnLr-i / t t j j t r j i . U n //, fr /s t* >T '-7i c c s - s . : j d / f e t n ? M - ... h s J z 4 j y ./ j^ > .W h JEDO..._______________ Actual Proceduirrec//pprrouGcCebs5s.: g. /jAc^- - .25/2- ________r_/_n__Z__,_/_____tT - c_ ^ ___z& o JL 3 /2 0 laJ ../ / V t J A s / l/ n s / n >>r?s77o^> ^ j b U i b y ' ^ j t 2 2 ^ ? J ____ .. / y j / a s ~ _________ III. A ctions Taken: ______________ (such a s am endm ent issued, S O P revision, e tc .)___________________ / s n e t? c /.j ? t# 2 L // Q J Recorded By Date f-Jc a dv<*A. IV. Im padt dfb&tudy / Project t,h OV/sx/i/ i>f>ctuir Mtrv Cott, i-tuda Dkitc^or CtQiTna L u e b & r K&CiA^AnCi-xC&GtL-tjb&^l / 3 M Environmental LaboratoryfC. 7~AE. t / b i b ) i b * / Deviation No. _______ 3_______ Form ETS-4-8.0 ` vt^H/ / (assigned by Study Director or Project Lead at the end o f study or project) 3 \I Environmental Laboratori' 001025 Page 29 418-018:PAGE 1-403 3M Medical Department Study: T-6295.25 3M Medical Department Study:T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-01BO Appendix C: Extraction and Analytical Methods This appendix includes the following methods: ETS-8-4.2, Extraction of Fluorochemical Compounds from Serum for Analysis Using HPLCElectrospray/Mass Spectrometry, (15 pages) ETS-8-5.2, Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry, (11 pages) 3M Environmental Laboratori' 001026 Page 30 3M Medical Department Study: T-6295.25 418-018:PAGE 1-404 Analytical Report: FACT-TOX-169 LRN-E01-0180 3M Environmental Laboratory M ethod E x t r a c t io n o f F l u o r o c h e m ic a l c o m po u n d s f r o m S e r u m f o r A n a l y sis U sin g H P L C -E l e c t r o sp r a y /M a ss Sp e c t r o m e t r y M ethod N um ber: E T S-8-4.2 A doption D ate: 03/01/99 E f f e c t iv e D a te : L<2-1 u 1 Approved By: Laboratory M anager -hU------------- - Group Leader o ^ /c j/ o/ D ate 0 'O -/ O 1 D ate 1.0 Scope and Application L i S co p e: This m ethod is for the extraction o f fluorochem ical com pounds from serum. 1.2 A p p lic a b le c o m p o u n d s : F lu oroch em ical surfactants or oth er flu orin ated com p o u n d s. ?& tl\(7 1.3 m X 0 M a tr ic e s : R abbit, rat, b o v in e, m onkey, and hum an serum or o th er fluids as d esig n a ted in the validation report. a 0 22..00 Summary of Method 1 02.1 This perform ance-based m ethod describes the procedure for extracting perfluorooctane- su lfon ate (P F O S ) or oth er flu oroch em ical surfactants from serum , o r o th er fluids, u sin g an ion pairing reagent and methyl-ierr-butyl ether (M tBE). An ion pairing reagent is added to the sam ple and the analyte-ion pair is partitioned into M tB E . T he M tB E extract is rem oved and put o n to a nitrogen evaporator until dry. Each extract is recon stitu ted in 1.0 m L o f m ethanol, then filtered through a 0 .2 pm nylon filter using a 3-m L plastic 3M Environmental Laboratori ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 1 of 15 001027 Page 31 3M Medical Department Study: T-6295.25 418-018:PAGE 1-405 Analytical Report: FACT-TOX-169 LRN-E01-0180 i 2.2 syringe into glass autovials. (A pplication o f this m ethod to seven fluorochem icals w as demonstrated: PFOS, PFO SA, PFO SAA, EtFO SE-O H , PFO SEA, M 556, and a surrogate standard (see 3 . 0 Definitions}). T hese sam ple extracts are analyzed follow ing m ethod E T S -8-5.2 or other appropriate m ethod. 3 .0 Definitions_______________________________________________________________ 3.1 PFOS: perfluorooctanesulfonate (anion o f potassium salt) CgF^SOj' 3 .2 P F O S A : p erflu o ro o cta n e sulfonylaxnide C 8F 17S 0 2N H 2 3 .3 P F O S A A : p erflu orooctan e su lfonylam ido (eth yl)acetate C gF i7S 0 2N (C H 2 C H 3)C H 2C 0 j ' 3 .4 EtFO SE-O H : 2(N -ethylperfluorooctane sulfonam ido)-ethyl alcohol C 8F i7S 0 2N (C H 2C H 3)C H 2C H 20 H 3 .5 P F O S E A : p erflu o ro o cta n e su lfon yl ethylam ide C 8F n S 0 2N (C H 2CH3)H 3 .6 M 5 5 6 : C 8F 17S 0 2N (H )(C H 2C O O H ) 3 .7 Surrogate standard TH PFO S: 1H -1H -2H -2H perfluorooctane sulfonic acid 4.0 Warnings and Cautions_____________________________ ;______________________ 4.1 Health and safety warnings 4.1.1 U se universal precautions, especially laboratory coats, goggles, and gloves when handling animal tissue, which may contain pathogens. 5.0 Interferences____________________________________________________________ 5.1 There are no interferences know n at this time. 6.0 Equipment _____________________________________________________________ 6.1 T he follow in g equipm ent is used w hile performing this m ethod. Equivalent equipm ent is acceptable. 6.1.1 V ortex m ixer, V W R , V ortex Genie 2 6 .1 .2 Centrifuge, M istral 1000 or IEC 6.1.3 Shaker, Eberbach or VW R 6 .1 .4 N itrogen evaporator, Organomation 6.1.5 Balance ( 0.100 g) 7.0 Su and Materials___________________________________________________ 7.1 G loves 7.2 E ppendorf or disposable pipettes, plastic or glass (or equivalent) 7.3 Polypropylene bottles, capable o f holding 250 mL and 1 L (N algene or equivalent) 3M Environmental Laboratory' ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 2 of 15 001028 Page 32 3M Medical Department Study: T-6295.25 418-018:PAGE 1-406 Analytical Report: FACT-TOX-169 LRN-E01-0180 7.4 Volum etric flasks, glass, type A 7.5 4 0 mL glass vials (I-CH EM or equivalent) 7.6 Centrifuge tubes, polypropylene, 15 mL 7.7 Labels 7.8 B ottle-top D ispenser - 3.0 to 10.0 mL or a 5 -1 0 mL graduated pipette, glass 7.9 Syringes, capable o f measuring 5 pL to 50 pL 7.10 Graduated pipettes 7.11 Syringes, disposable plastic, 3 mL (B& D or equivalent) 7.12 Syringe filters, nylon, 0.2 pm , 25 mm 7.13 Timer 7.14 Crimp cap glass autovials and caps 7 .15 Crimpers N ote: Prior to using glassw are and bottles, rinse 3 times with m ethanol and 3 tim es with water. R inse syringes a minimum o f 9 tim es with m ethanol, 3 rinses from 3 separate vials. 8 .0 Reagents and Standards__________________________________________________ 8.1 R eagent grade w ater, M illi-Q TM , Nanopure II or equivalent 8.2 Sodium hydroxide (N aO H ), J.T Baker or equivalent 8.3 Tetrabutylammonium hydrogen sulfate(TBA), Kodak or equivalent 8.4 S odium carbonate (Na^COg), J.T. Baker or equivalent 8.5 Sodium bicarbonate (NaH CO a), J.T. Baker or equivalent 8.6 M ethyl-T -B utyl Ether, O m nisolv, glass distilled or HPLC grade 8.7 M ethanol, O m nisolv, glass distilled or HPLC grade 8.8 Serum or blood, frozen from supplier 8.9 F luorochem ical standards 8.9.1 KPFOS (PFO S) (3M Specialty Chemical Division), m olecular w eight = 538 8 .9 .2 PFOS A (3M Specialty Chemical D ivision), molecular w eight = 499 8 .9 .3 PFO SA A +H (PF O SA A ) (3M Specialty Chemical D ivision), m olecular w eight = 585 8 .9 .4 EtFO SE-O H (3M Specialty Chemical D ivision), m olecular w eight = 570 8 .9 .5 PFO SE A (3M Specialty Chemical D ivision), molecular w eight = 527 8.9.6 M 556 (3M Specialty Chemical D ivision), molecular w eight = 557 8 .9 .7 S u rrogate standard: 4 -H , p erflu orooctan e su lfon ic acid (1 -H , 1-H , 2 -H , 2 -H C gFuSO H , [TH PFO S]), molecular weight = 428 8 .9 .8 Other fluorochem icals, as appropriate 3M Environmental Laboratori' ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 3 of 15 001029 Page 33 3M Medical Department Study: T-6295.25 418-018:PAGE 1-407 Analytical Report: FACT-TOX-169 LRN-E01-0180 8.10 8.11 R eagent preparation N O T E : W h en preparing different v o lu m es than listed in reagen t, standard, o r surrogate preparation, adjust accordingly. 8.10.1 IO N sodium hydroxide (NaOH): W eigh approximately 2 0 0 g N aO H . Pour into a 1000 m L beaker containing 5 0 0 mL water, m ix until all solids are dissolved. Store in a 1 L polypropylene (or equivalent) bottle. 8 .1 0 .2 1 N sodium hydroxide (NaO H ): D ilute 10 N N aO H 1:10. M easure 10 raL o f 10 N N aO H solution into a 100 mL volum etric flask and bring to volum e using water. Store in a 125 m L polypropylene (or equivalent) bottle. 8 .10.3 0.5 M tetrabutylammonium hydrogen sulfate (TBA): W eigh approxim ately 169 g o f T B A into a 1 L volum etric containing 500 mL water. Adjust to pH 10 using approxim ately 4 4 to 54 m L o f 10 N N aO H . (A dd the last 1.0 m L o f N aO H slow ly, as the pH changes abruptly.) D ilute to volum e w ith w ater. S tore in a 1 L polypropylene (or equivalent) bottle. 8 .1 0 .3 .1 T B A requires a check prior to each use to ensure pH = 10. Adjust as needed using 1 N N aO H solution. 8 .1 0 .4 0 .2 5 M so d iu m carb on ate/sod iu m bicarbonate buffer (N a jC O j/N a H C 0 3): W eigh ap proxim ately 2 6 .5 g o f so d iu m carbonate (N a 2C O j) and 2 1 .0 g o f so d iu m bicarbonate ( N a H C 0 3) into a 1 L volum etric flask and bring to v o lu m e w ith w ater. Store in a 1 L polypropylene (or equivalent) bottle. Standards preparation 8 .1 1 .1 Prepare PFOS standards for the standard curve. 8 .1 1 .2 Prepare other fluorochem ical standards, as appropriate. M ulticom ponent fluorochem ical standards are acceptable (for exam ple, one working standard solution containing approximately 1.00 pg/m L o f PFO S, PFOS A , PFO SA A , PFO SEA , M 556 and EtFOSE-OH). 8 .1 1 .3 W eigh approxim ately 100 m g o f PFOS into a 100 m L volum etric flask and record the actual w eig h t. F o r standards w ith KT or other salts, m ultiply b y a correction factor. For exam p le, the m olecular w eight o f KPFOS = 5 3 8 , and the m olecular w eight o f PFOS = 499. Calculate the correction factor by using the follow ing equation: m olecular wt. PFOS ( 499 ) m olecular wt. KPFOS (538) 0.9275 ( Correction factor ) 8 .1 1 .4 Bring to volum e w ith m ethanol for a stock standard o f approxim ately 1000 pg/m L. 8 .1 1 .5 D ilute the stock solution w ith methanol for a working standard 1 solution o f approximately 5 0 pg/m L. lQ 0 0 fig /m L x 5 m L = 50lglmL 100 mL 8.11.6 D ilu te w ork in g standard 1 w ith m ethanol fo r a w o rk in g standard 2 so lu tio n o f approx. 5.0 pg/m L. 3.\f Environmental Laboratory ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 4 of 15 001030 Page 34 3M Medical Department Study: T-6295.25 418-018:PAGE 1-408 A nalytical Report: FA C T -T O X -169 L R N -E 01-0180 f 5 0 /g / m L x .lQ m L ,'' = 5.0 lig /m L { 100/nL ; 8 .1 2 8 .1 1 .7 D ilute working standard 1 with methanol for a working standard 3 solution o f approx. 0.50 pg/m L. 50Hg t mL x 1.0mL =0.500 fig ImL 100 mL Surrogate T H PFO S stock standard preparation 8.12.1 W eigh approxim ately 5 0 -6 0 m g o f surrogate standard 1 -H .l-H , 2-H , 2-H , CgFnSO sH (TH PFO S) into a 50 mL volum etric flask and record the actual w eight. 8 .1 2 .2 Bring to volum e with m ethanol for a surrogate stock o f approxim ately 10001200 pg/m L. 8 .1 2 .3 Prepare a surrogate working standard. Transfer approximately 1 m L o f surrogate stock to a 10 m L volum etric flask and bring to volum e with m ethanol for a working standard o f 100 pg/m L. Record the actual volum e transferred. 9 .0 Sample Handling_________________________________________________________ 9.1 A ll sam ples are received frozen and m ust be kept frozen until the extraction is perform ed. 9 .2 A llo w sam p les to thaw at r o o m tem perature or in lukew arm w ater prior to extraction . 10 .0 Quality Control_______________________________________________________ 10.1 S o lv en t B lan k s, M eth od B lan k s a n d M atrix B lanks 1 0 .1 .1 Solvent Blanks: A n aliquot o f 1.0 m L m ethanol is used as a solvent blank. 1 0 .1 .2 M ethod Blanks: Follow ing this procedure, extract tw o 1.0 m L aliquots o f water and use as m ethod blanks. 10.1.3 Matrix Blanks: Matrix blanks are prepared from one of three sources: l) a study control matrix from a study control animal received with each sam ple set; 2) a com m ercially obtained sam ple o f the sam e species as the study animals; or 3) a surrogate matrix, also obtained com m ercially, but o f a different species than the study animal (e.g . if rabbit is used to generate standard curves and C C V s for a m onkey or rat sera study). The matrix to use depends on what matrix is used for the curve. 1 0 .1 .3 .1 Study control matrix curve-- I f the study control matrix is used for the curve, prepare tw o (2) matrix blanks using the study control matrix. 1 0 .1 .3 .2 C om m ercially obtained (sam e sp ecies) matrix curve-- I f the curve is prepared using com m ercially obtained matrix in the sam e sp ecies as the stu d y anim al, prepare tw o matrix blanks u sin g this; sam e com m ercially available matrix. 3M Environmental Laboratori' ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 5 of 15 001031 Page 35 3M Medical Department Study: T-6295.25 418-018:PAGE 1-409 Analytical Report: FACT-TOX-169 LRN-E01-0180 10.1.3.3 Surrogate matrix curve--If a surrogate matrix is used for the curve and continuing calibration verification samples: a) prepare two (2) matrix blanks using the surrogate matrix. b) Also, prepare a matrix blank with each set of matrix spikes (a set is a matrix spike/matrix spike duplicate at one level) using a commercially available matrix of the same species as the study animals. 10.2 Matrix spikes 10.2.1 Study Control Matrix Curve No matrix spikes are prepared. 10.2.2 Commercially obtained matrix curve (same species) No matrix spikes are prepared. 10.2.3 Surrogate matrix curve Matrix spikes are necessary if matrix is not available in the same species as the study animal and a surrogate matrix is used for the curve and CCV samples (e.g., rabbit sera may be used to generate standard curves for a monkey sera study, due to measurable levels of endogenous analyte in the monkey sera). Prepare and analyze matrix spike and matrix spike duplicate samples to verify the accuracy of the extraction for target analytes. 10.2.4 Prepare each MS and MSD at two (2) levels (usually a low and mid-range concentration) using a sample chosen by the analyst, typically sera from a control group animal received with each sample set. 10.2.4.1 If there is not sufficient sera available from the control group, prepare matrix spikes using commercially available matrix from the same species as the study animal. 10.2.5 Prepare one matrix spike and matrix spike duplicate at each level listed in 10.2.4 per 20 samples, with a minimum of 2 matrix spikes per level per batch. If a batch includes more than 20 samples, additional spikes may be prepared at the same, low, or high range levels. 10.2.6 If more than 25% of the samples are <LOQ, two matrix spikes should be prepared at approximately 2-5 times the expected LOQ. 10.2.7 If the majority of the samples are at or above the high range of the curve, additional matrix spikes should be prepared to approximate sample concentrations. 10.3 Continuing calibration verifications (CCVs) 10.3.1 Prepare continuing calibration verification samples for instrument stability verification during analysis. Prepare and analyze CCVs for every assay run, regardless of the matrix type used to prepare the standard curve. 10.3.2 Prepare each continuing calibration verification from the same matrix used to prepare the initial curve (i.e., either the study control matrix, commercial matrix of same species, or surrogate matrix). SM Environmental Laboratori' ETS-8-4.2 Extraction of Fluorochcmicals from Serum Page 6 of 15 001032 Page 36 3M Medical Department Study: T-6295.25 418-018:PAGE 1-410 Analytical Report: FACT-TOX-169 LRN-E01-0180 10.3.3 The expected concentrations of the CCVs will fall within the range of the initial calibration curve; typically, the low to mid-range of the curve. 10.3.4 Prepare, at a minimum, two continuing calibration verifications, each at a different concentration, per group of 10 samples. For example, if a sample set = 34, eight (8) checks are prepared; four (4) at a low range and four (4) at a mid-range concentration. 10.3.5 Additional continuing calibration verifications may be included at the same, low, mid or high-range concentration. 11.0 Calibration and Standardization_________________________________ 11.1 Prepare matrix calibration standards 11.1.1 Transfer 1mL of appropriate serum to a 15 mL centrifuge tube. If commercially available sera is used, prepare two matrix blanks, using the volume of sera available for most study animals. 11.1.1.1 Depending upon study goals or if analyte-free commercial sera is not available, a surrogate matrix may be used for generation of the calibration standard. For example, rabbit sera may be used as a surrogate matrix for rat studies if quantitation of extremely low levels of the analyte is required. 11.1.2 If most sample volumes are less than 1.0 mL, extract standards with matrix volumes equal to the sample volumes. Do not extract less than 0.50 mL of matrix. Record each sample volume on the extraction sheet. 11.1.3 While preparing a total of thirteen aliquots in 15-mL centrifuge tubes, mix or shake sera between aliquots. 11.1.4 Two 1-mL aliquots, or other appropriate volume, serve as curve matrix blanks. Typically use the standard concentrations and spiking amounts listed in Table 1at the end of this section to spike one standard curve, for a total of nine standards, two matrix blanks, and two method blanks. 11.1.5 Refer to validation report ETS-8-4.0 & ETS-8-5.0-V-1, which lists the working ranges and the Linear Calibration Range (LCR) for calibration curves. 11.1.6 See Section 13.0 to calculate actual concentrations of PFOS in calibration standards. 11.2 To each standard, blank, continuing check, and sample add an appropriate amount of surrogate working standard to achieve a constant concentration :hat falls within the calibration curve range of 2.5 ng/mD-1000 ng/mL. Usually, samples are spiked for a constant concentration in all samples at approximately 500 ng/mL or other concentration as determined by the analyst. 11.3 Extract spiked matrix standards following 12.6-12.16 of this method. Use these standards to establish each initial curve on the mass spectrometer. SM Environmental Laboratory' ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 7 of 15 001033 Pa8e3? 3M Medical Department Study: T-6295.25 418-018:PAGE 1-411 Analytical Report: FACT-TOX-169 LRN-E01-0180 T able 1 A pproxim ate spiking am ounts for standards and spikes U sin g 1.0 m L o f m atrix W orking standard (approx, cone.) Approx, final con e, o f \lL analyte in matrix - - Blank 0.500 ppm 10 0.005 ppm 0.500 ppm 20 0.010 ppm 5.00 ppm 5 0.025 ppm 5.00 ppm 10 0.050 ppm 5.00 ppm 20 0.100 ppm 50.0 ppm 5 0.250 ppm 50.0 ppm 10 0 .5 0 0 ppm 50.0 ppm 15 0 .7 5 0 ppm 50.0 ppm *ppm=pg/mL 20 1.00 ppm 12.0 Procedure_______________________________________________________________ 12.1 Obtain frozen samples and allow to thaw at room temperature or in lukewarm water. 12.2 Label a 15 mL polypropylene centrifuge tube with the study number, sample ID, date and analyst initials. See attached worksheet (Attachment A or similar worksheet) for documenting the remaining steps. 12.3 Vortex mix for 15 seconds, then transfer 1.0 mL or other appropriate volume to the 15 mL polypropylene centrifuge tube. 12.4 Return unused samples to freezer after extraction amounts have been removed. 12.5 Record the initial volume on the extraction worksheet (Attachment A or similar worksheet). 12.6 Spike all samples, blanks and standards, that are ready for extraction with surrogate standard as described in 11.2. 12.7 Spike each calibration standard matrix with the appropriate amount of standard as described in 11.1, or Table 1 in that section, for the calibration curve standards. Also prepare matrix spikes if necessary and continuing calibration standards. 12.8 Vortex mix the standard curve samples, matrix spike samples, and continuing calibration samples for 15 seconds. 12.9 Check to ensure the 0.5 M TBA reagent is at pH 10. If not, adjust accordingly. 12.10 To each sample, add 1 mL 0.5 M TBA and 2 mL of 0.25M sodium carbonate/sodium bicarbonate buffer. 12.11 Using an bottle top dispenser or 5-10 mL graduated glass pipette, add 5 mL methyl-ierrbutyl ether. 12.12 Cap each sample and put on the shaker at a setting of 300 rpm for 20 minutes. 3M Environmental Laboratory- ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 8 of 15 001034 Page 38 3M Medical Department Study: T-6295.25 418-018:PAGE 1-412 Analytical Report: FACT-TOX-169 LRN-E01-0180 12.13 Centrifuge for 20 to 25 minutes at a setting of 3500 rpm or until layers are well separated. 12.14 Label a fresh 15 mL centrifuge tube with the same information as in 12.5. 12.15 Remove 4.0 mL of the organic (top) layer to this clean 15 mL centrifuge tube. 12.16 Put each sample on the analytical nitrogen evaporator until dry, approximately 1 to 2 hours. 12.17 Add 1.0 mL of methanol to each centrifuge tube using a graduated pipette. 12.18 Vortex mix for 30 seconds, or longer if needed. 12.19 Label a 1.5 mL glass autovial (or low-volume autovial when necessary) with the study number, animal number and gender, sample timepoint, matrix, final solvent, extraction date, and analyst(s) performing the extraction. 12.20 Attach a 25 mm, 0.2 pm nylon mesh filter to a 3 mL syringe and transfer the sample from step 12.18 to this syringe. Filter into the labeled autovial. 12.21 Cap and store extracts at room temperature or at approximately 4 C until analysis. 12.22 Complete the extraction worksheet (Attachment A or similar worksheet) and include in the study binder. 13.0 Data Analysis and Calculations__________________________________________ 13.1 Calculations 13.1.1 Calculate actual concentrations of PFOS, or other applicable fluorochemical, in calibration standards using the following equation: _________ m L o f std x concentration o f std ( fig / m L )_________ mL o f std + mL o f surrogate std + initial matrix volume ( m L ) Final concentration ( ftg / mL ) o f PFOS in matrix 14.0 M e t h o d P e r f o r m a n c e ________________________________________________________________ 14.1 The method detection limit (MDL) is analyte and matrix specific. Refer to MDL report for specific MDL and limit of quantitation (LOQ) values (see Attachments B and C). At the discretion of the PAI, MDL may not be defined for some studies. 14.2 The following quality control samples are extracted with each batch of samples to evaluate the quality of the extraction and analysis. 14.2.1 Method blanks and matrix blanks. 14.2.2 If surrogate matrix is used for curve and QC, matrix spike and matrix spike duplicate samples to verify extraction efficiency. 14.2.3 Continuing calibration check samples to determine the continued accuracy of the initial calibration curve. 14.3 Refer to section 14 of ETS-8-5.2 for method performance criteria. 3M Environmental Laboratori' ETS-8-4.2 Extraction of Fluorochcmicals from Serum Page 9 of 15 001035 Page 39 3M Medical Department Study: T-6295.25 418-018:PAGE 1-413 Analytical Report: FACT-TOX-169 LRN-E01-0180 15.0 Pollution Prevention and Waste Management_______________________ 15.1 Dispose of sample waste, flammable solvent waste and used glass pipette waste using proper methods and by placing each in their designated waste container. 16.0 Records_________________________________________________________________ 16.1 Complete the extraction worksheet attached to this method (or a similar worksheet), and include in the study binder. 17.0 Attachments____________________________________________________ _ 17.1 Attachment A, Extraction worksheet (a similar worksheet may be substituted for Attachment A) 17.2 Attachment B, MDL/LOQ values and summary 17.3 Attachment C, Ion Pair Standard Curves worksheet (a similar worksheet may be substituted for Attachment C) 18.0 References_________________________________________________________ 18.1 The validation report associated with this method is ETS-8-4.0 & 5.0-V-l. 18.2 FACT-M-3.1, "Analysis of Serum or Other Fluid Extracts for Fluorochemicals using HPLC-Electrospray Mass Spectrometry" 18.3 ETS-8-5.2, "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" 19.0 Affected Documents_____________________________________________________ 19.1 ETS-8-5.2, "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" 20.0 Revisions_______________________________________________________________ Revision Number 1 2 Reason For Revision Section 12.21 Changed to include sample storage at room temperature. Section 12.13 Added the shaker speed. Section 12.17 Final volume is 1.0 mL; not adjusted for initial volumes less than 1.0 mL. Section 1.1, 2.1 Eliminate `potassium' from perfluorooctanesulfonate 8.9.1 Add K to PFOS 8.9.3 Add H to PFOSAA 10.2-10.2.4, 14.2.2 Add information about using surrogate matrix, clarify control matrix and spiking the curve, change spikes to every 20 samples 10.3 Clarify purpose of continuing calibration checks 11.1 Added wording about control animal sera, using and not using commercially available tissue In general, make less specific for equipment/supplies. Substitute water for Milli-Q, Nalgene or equivalent bottle may be used. Revision Date 04/02/99 3M Environmental Laboratory ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 10 of 15 001036 Page 40 418-018:PAGE 1-414 3M Medical Department Study: T-6295.25 A nalytical Report: FA C T -T O X -169 Extraction Worksheet ETS-8-4.2 L R N -E 01-0180 Studv # Matrix Box # Wk/Dav Date Spiked/ Analyst: Surrogate Std actual Mg/mL # FC-Mix approx. 0.5 Mg/mL # FC-Mix approx. 5 Mg/mL # FC-Mix approx. 50 Mg/mL # Comments Verified by: MS MSD -----.- - -- -- - -- - -- - -- - -- - -- - -- - -- - Blank___________ OEL2___________________________________ amount =________________mL___________ -Simm-ExtractionMfilhod_________________________ :................................... . Vortex 15 sec. Date & initials 1 1 Pi,agtieJd?ma>5fflLlsE-yilti appropriai surrogale or FCMix_____ -..... volume Pioette 1 mL of 0.5 M TBA oH 10. pH = Std. # mL Pipette 2 mL of 0.25 NajCOV0.25M NaHCO, buffer Std. # Dispense/pioette 5 mL of methvl-t-butvl ether TN-A- Dispenser ID #: Shake 20 min. Shaker ID #: Shaker speed: -Cauiifiisc 2Q-2 mio.____ C a u riftg cB f;________________________ Qatrifugg spcgfli- Rcmpyg,,a 4.mL aliaupt of organic lavec---------------------------------------------------------------- ...Pulon MitregcnEyawraiot to dryness------------------------------------------------------------------ Add methanol Volume mL TN-A- VfiUCXJO 5fC. Filter usine a 3mL B-D svrinee with a 0.2um filter into a 1.5 mL auiosamole vial_________ Com. Cal. Verifications used same matrix as for std curve. Appendix A: Extraction Worksheet 3M Environmental Laboratory ETS-8-4.2 Page 11 of 15 001037 Page 41 3M Medical Department Study: T-6295.25 418-018:PAGE 1-415 A nalytical Report: FA C T -T O X -169 L R N -E 01-0180 MDL/LOO values for rabbit serum Compound MDL LOQ Linear Calibration Range (LCR) (ng/mL) (ng/mL) Approximate concentrations to be used for preparing the Standard Calibration Curve PFOS 1.74 5.55 5 ng/mL-1000 ng/mL PFOSA 1.51 4.79 5 ng/mL-1000 ng/mL PFOSAA 3.46 20.5 5 ng/mL-1000 ng/mL EtFOSE-OH 11.4 36.2 5 ng/mL-1000 ng/mL M556 6.03 19.2 5 ng/mL-1000 ng/mL PFOSEA 5.71 18.2 5 ng/mL-1000 ng/mL MDULOQ values in rat, bovine, monkey, and human serum, and monkey plasma were not statistically determined. Two curves in each of these matrices were extracted and analyzed with the rabbit serum curves to determine equivalence. Responses in the rat, bovine, monkey, and human were equivalent to the rabbit responses, therefore, their MDL and LOQ will be the same values as determined in rabbit serum. Please see LOQ Summary and MDL study in ETS-8-4.0 & 5.0-V-l for further information. Appendix B: MDL7LOQ Values and Summary ETS-8-4.2 Extraction of Fluorochemicals from Serum 3M Environmental Laboratory- Page 12 of 15 001038 Page 42 3M Medical Department Study: T-6295.25 418-018.PAGE 1-416 Analytical Report: FACT-TOX-169 LRN-EOl-0180 Compound: PFOS Prepared range Rabbit Scrum of standards (ng/mL) Full Range 0.995 - 978 Low Curve 4.94 - 248 High curve 97.8 - 978 1X 0.995 - 978 Compound: PFOSA Prepared range Rabbit Serum o f standards (ng/mL) Full Range 0.993-976 Low Curve 4.93-97.6 High curve 24.8-976 1/X 0.993-976 Com pound: F FOSAA Prepared range Rabbit Serum o f standards (ng/mL) Full Range 0.991 -974 Low Curve 4.92-247 High curve 49.2-974 1/X 0.991 -974 LCR from curve (ng/mL) 24.8 - 978 4.94 - 248 97.8 - 978 4.94 - 978 LCR from curve (ng/mL) 4.93-976 4.93 -97.6 24.8-978 4.93-976 LCR from curve (ng/mL) 24.7-974 9.74-247 97.4-974 9.74-974 % Recovery Range 83-108 85-104 85-106 94-111 % Recovery Range 88-103 87-105 93-102 94-103 % Recovery Range 81-111 97-107 85-108 95-115 RSD Range 4.67-11.0 5.34-12.0 4.84-9.80 4.60-10.5 RSD Range 5.10-14.7 9.85-14.7 5.08-13.9 5.10-14.5 RSD Range 4.18-10.6 6.38-21.8 4.33-12.5 4.11-23.2 Appendix B: MDL/LOQ Values and Summary ETS-8-4.2 Extraction of Fluorochemicals from Serum 3M Environmental Laboratory Page 13 of 15 001039 Page 43 3M Medical Department Study: T-6295.25 418-018.PAGE 1-417 A nalytical Report: FA C T -T O X -169 L R N -E 01-0180 Compound: EtFQSE-OH Prepared range Rabbit Serum of standards (ng/mL) Full Range 0.993 - 976 Low Curve 4.93 - 97.6 High curve 49.3 - 976 1/X 0.993 - 493 Compound: PFOSEA Prepared range Rabbit Serum of standards (ng/mL) Full Range 0.993 - 976 Low Curve 4.93 - 248 High curve 49.3 - 976 1/X 0.993 - 976 Compound: M556 Prepared range Rabbit Serum of standards (ng/mL) Full Range 0.993 - 976 Low Curve 4.93 - 97.6 High curve 97.6 - 976 1/X 0.993 - 976 LCR from curve (ng/mL) 49.3 - 976 9.76-97.6 97.6 - 976 9.76 - 976 LCR from curve (ng/mL) 24.8 - 976 9.76 - 248 49.3 - 976 9.76 - 976 LCR from curve (ng/mL) 24.8 - 976 9.76 - 97.6 97.6 - 976 9.76 - 976 % Recovery Range 77-110 97-107 90-109 8 6 -111 % Recovery Range 96-106 91-110 86-106 95-117 % Recovery Range 88-106 100-105 81-111 97-110 RSD Range 11.2-25.5 14.1-21.3 11.5-19.6 1 1 . 1 -2 1 . 2 RSD Range 10.1-16.2 11.8-19.5 10.2-18.2 10.1-19.1 RSD Range 4.82-17.9 5.95-18.2 5.11-9.74 4.77-19.5 Appendix B: MDL/LOQ Values and Summary ETS-8-4.2 Extraction of Fluorochemicals from Scrum 3M Environmental Laboratori Page 14 of 15 001040 Page 44 3M Medical Department Study: T-6295.25 418-018:PAGE 1-418 Analytical Report: FACT-TOX-169 LRN-E01-0180 E xam ple Ion Pair Standard Curves - Fluids Analyses) i Prep date(s): Study number: Final solvent and TN: Analyte(s): Study Animal M atrix: Blank fluid/identifier: !R*>rd matrix used to reran- curve-. ;.o.e.. `Rahi^!u' ID *X X X }| Method/revision: Target analyte(s): FC mix std approx. 0.500 ppm: FC mix std approx. 5.00 ppm: FC mix std approx. 50.0 ppm; [ Record the standards used lu prepari- Ilk- oLtraeied ctiru-s. | Surrogate std approx. 100 ppm: Actual concentrations of standards in the FC mix in methanol PFOS PFOSA PFOSAA EtFOSE PFOSEA M556 Std cone Std cone Std cone Std cone Std cone Std cone pg/mL pg/mL Mg/mL pg/mL Hg/mL Mg/mL 0.500 0.507 0.532 0.501 0.521 0.501 0.500 0.507 0.532 0.501 0.521 0.501 5.00 5.07 5.32 5.01 5.21 5.01 5.00 ^1 f 5D7 5.32 5.01 5.21 "1 5.01 5.00 B-- 50.0 1 ! 50.1 J 50.0 S L sat1 / I 3.1 1 | 5 2 l 1 o n Jl A i JL 50.0 50.1 53.2 50.1 R 52.1 50.1 50.0 50.1 53.2 50.1 52.1 50.1 All Ain't spiked mL 0 .0 10 0 .0 20 0.005 0 .0 10 0 .0 20 0.005 0 .0 10 0.015 0 .0 20 All Final voi mL 1.015 1.025 1.0 10 1.015 1.025 1.0 10 1.015 1.0 2 0 1.025 Calculated concentrations of standards in the sera matrix: Rabbit PFOS PFOSA PFOSAA EtFOSE PFOSEA M5S6 Surrogati: Final cone Final cone Final cone Final cone Final cone Final cone Std cone ng/mL ne/mL ng/mL ng/mL ng/mL ng/mL ne/mL AU Amt spiked mL Validated ranges - approximate concentrations Serum PFOS PFOSA PFOSAA Rabbit 5.00-1000 5.00-1000 5.00-1000 ng/mL ng/mL ng/mL Bovine Estimates only. Use values for rabbit. Rat Estimates only. Use values for rabbit. Monkey & Plasma Estimates only. Use values for rabbit. Human Estimates only. Use values for rabbit. EtFOSE-OH 5.00-1000 ng/mL PFOSEA 5.00-1000 ng/mL M5S6 5.00-1000 ng/mL Attachment C: Ion Pair Standard Curves ETS-8-4.2 Extraction of Fluorochcmicals from Serum 3M Environmental Laboratory Page 15 of 15 001041 Page 45 3M Medical Department Study: T-6295.25 418-018:PAGE 1-419 A nalytical Report: F A C T -T O X -169 L R N -E 01-0180 3M Environmental Laboratory Method Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/M ass Spectrometry Method Number: ETS-8-5.2 Author: Lisa Clemen, Kris Hansen Approved By: Laboratory Manager Adoption Date: 03/01/99 Revision Date: I 110 1 O /cJ/ Date / Group Leader Technical Reviewer Date ?1_ - o (. o ( Date Exact Copy 1.0 Scope and Application____________________________________________________ 1.1 Scope: This method describes the analysis of serum extracts for fluorochemical surfactants using HPLC-electrospray/mass spectrometry. 1.2 Applicable Compounds: Fluorochemical surfactants or other fluonnated compounds, or other ionizable compounds. 1.3 Matrices: Rabbit, rat, bovine, monkey, and human serum, or other fluids as designated in the validation report. * Word 6/95 3M Environmental Laboratory ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 1 of 11 001042 Page 46 3M Medical Department Study: T-6295.25 418-018:PAGE 1-420 Analytical Report: FACT-TOX-169 LRN-E01-0180 2.0 Summary of Method____________________________ __________________________ 2.1 Although supported by a validation for most commonly used matrices, this is a performance-based method. Careful attention should be paid to method QC as there is great variability in sera. This method describes the analysis of fluorochemical surfactants extracted from serum or other fluids, using HPLC-electrospray/mass spectrometry, or similar system as appropriate. The analysis is performed by monitoring a single ion characteristic of a particular fluorochemical, such as the perfluorooctanesulfonate (PFOS) anion, m/z= 499. Additionally, samples may be analyzed using a tandem mass spectrometer to further verify the identity of a compound by detecting daughter ions of the parent ion. 3.0 Definitions_______________________________________________ _ 3.1 Atmospheric Pressure Ionization (API): The Micromass Quattro II and Ultima triple quadrupole systems allow for various methods of ionization by utilizing various sources, probes, and interfaces. These include but are not limited to: Electrospray Ionization (ESI), Atmospheric Pressure chemical Ionization (APcI), Thermospray, etc. The ionization process in these techniques occurs at atmospheric pressure (i.e., not under a vacuum). 3.2 Electrospray Ionization (ES, ESI): a method of ionization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These charged droplets are produced by the application of a strong electrical field. 3.3 Mass Spectrometry, Mass Spectrometer (MS), Tandem Mass Spectrometer (MS/MS): The API Quattro II and Ultima triple quadrupole systems are equipped with quadrupole mass selective detectors. Ions are selectively discriminated by mass to charge ratio (m/z) and subsequently detected. A single MS may be employed for ion detection or a series (MS/MS) for more specific fragmentation information. 3.4 Conventional vs. Z-spray probe interface: The latest models of Micromass triple quadrupole systems (post 1998) utilize a "Z-spray" conformation. The spray emitted from a probe is orthogonal to the cone aperture. In the conventional confonnation it is aimed directly at the cone aperture, after passing through a tortuous pathway in the counter electrode. Though the configuration is different, the methods of operation, cleaning, and maintenance are the same. However, Z-spray components and conventional components are not compatible with one another, but only with similar systems (i.e., Z-spray components are compatible with some other Z-spray systems, etc.) 3.5 Mass Lynx Software: System software designed for the specific operation of these Quattro II triple quadrupole systems. Currently MassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details see the manual specific to the instrument (Micromass Quattro II or Ultima triple quadrupole MassLynx or MassLynx NT User's Guide). 4.0 Warnings and Cautions______________________________________________ 4.1 Health and Safety Warnings: 4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe employs a voltage of approximately 5000 Volts. Word 6/95 5M Environmental Laboratory ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 2 of 11 001043 Page 4 7 3M Medical Department Study: T-6295.25 418-018:PAGE 1-421 Analytical Report: FACT-TOX-169 LRN-E01-0180 4.1.2 When handling samples or solvents wear appropriate protective gloves, eyewear, and clothing. 4.2 Cautions: 4.2.1 Do not operate solvent pumps above capacity of 400 bar (5800 psi) back pressure. If the back pressure exceeds 400 bar, the HPLC will initiate automatic shutdown. 4.2.2 Do not run solvent pumps to dryness. 5.0 Interferences___________________________________________________________ 5.1 To minimize interferences when analyzing samples, Teflon should not be used for sample storage or any part of instrumentation that comes in contact with ihe sample or extract. 6.0 Equipment______________________________________________________ 6.1 Equipment listed below may be modified in order to optimize the system. Document any modifications in the raw data as method deviations. 6.1.1 Micromass Quattro II or Ultima triple quadrupole Mass Spectrometer equipped with an electrospray ionization source 6.1.2 HP1100 or Agilent low pulse solvent pumping system, solvent degasser, column compartment, and autosampler 7.0 Supplies and Materials___________________________________________________ 7.1 Supplies 7.1.1 High purity grade nitrogen gas regulated to approximately 100 psi (House air or nitrogen system) 7.1.2 HPLC analytical column, specifics to be determined by the analyst and documented in the raw data. 7.1.3 Capped autovials and capped 15 mL centrifuge tubes 8.0 Reagents and Standards_________________________________________________ 8.1 Reagents 8.1.1 Methanol, HPLC grade or equivalent 8.1.2 Milli-QTM water, all water used in this method should be Milli-QTM water or equivalent, and may be provided by a Milli-Q TOC Plus system or other vendor 8.1.3 Ammonium acetate, reagent grade or equivalent 8.2 Standards 8.2.1 Typically two method blanks, two matrix blanks, and eighteen matrix standards are prepared during the extraction procedure. See ETS-8-4.2. 3M Environmental Laboratory' ETS-8-S.2 Analysis of Serum Extract Using ES/MS Page 3 of 11 001044 Page 48 3M Medical Department Study: T-6295.25 418-018-.PAGE 1-422 Analytical Report: FACT-TOX-169 LRN-E01-0180 9.0 Sample Handling____________________________________ ____________________ 9.1 Fresh matrix standards are prepared with each analysis. Extracted standards and samples are stored in capped autovials or capped 15 mL centrifuge tubes until analysis. 1 9.2 If analysis will be delayed, extracted standards and samples can be refrigerated at approximately 4 C, or at room temperature, until analysis can be performed. 10.0 Quality Control_________________________________________ 10.1 Solvent Blanks, Method Blanks and Matrix Blanks ,________ 10.1.1 Solvent blanks, method blanks and matrix blanks are prepared and analyzed at least once during the course of the study to determine if contamination occurred during sample prep. 10.1.2 Analyze at least one solvent blank prior to each calibration curve. 10.1.3 Matrix blanks should be analyzed with each sample list that includes undiluted extracts. 10.2 Matrix Spikes 10.2.1 If curves and method QC are prepared in a surrogate matrix (e.g. curves in rabbit sera, samples are monkey sera), matrix spikes and matrix spike duplicates are prepared in blank sample matrix (e.g., monkey sera) and analyzed to verify extraction efficiency. 10.2.2 If curves and method QC are prepared in the same matrix as samples, no additional matrix spikes are required. 10.3 Continuing Calibration Verifications (CCVs) 10.3.1 Continuing calibration verifications are analyzed to verify the continued accuracy of the calibration curve. 10.3.2 Analyze two calibration standards (one at each of 2 levels) after every one to ten samples, with a minimum of two per batch and always finishing an injection sequence with at least two calibration standards. 11.0 Calibration and Standardization_________________________________________ 11.1 Analyze the extracted matrix calibration standards prior to each set of extracts. The curve will be plotted by linear regression, weighted 1/x, not forced through zero, using MassLynx or other suitable software. 11.2 If the curve does not meet requirements, perform routine mainteniince, reextract samples, or reanalyze the standard curve. 11.3 For purposes of accuracy when quantitating levels of analyte at the limits of the curve range, it may be necessary to use either the low end or the high end of the calibration curve rather than the full range of the standard curve. Example: when attempting to quantitate approximately 10 ppb of analyte, it may be beneficial to generate a calibration curve consisting of the standards from 5 ppb to 100 ppb rather than the full range of the curve (5 ppb to 1000 ppb). This will reduce inaccuracy attributed to linear regression weighting of 3 \f Environmental Laboratori.' ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 4 of 11 001045 Page 49 3M Medical Department Study: T-6295.25 418-018:PAGE 1-423 Analytical Report: FACT-TOX-169 LRN-E01-0180 high concentration standards. It is also acceptable to break the linear range into a low curve and a high curve. If this is done, no more than one point should be used in common between the curves. For example, the low curve may include the following points: 1, 5, 25, 100, 250 ppb and the high curve may include the points: 250, 500, 750, 1000, 1250 ppb. 12.0 Procedures__________________________________________________________ _ 12.1 Acquisition Set up 12.1.1 On the MassLynx main page, set up a sample list name. Save the list as instrument designator letter, last 2 digits of test year-mo-day, and a letter that will increase through the alphabet with each additional list for that day. Example Sample List: IYYMMDDa or DO10712a I=instrument name (D for "Davey") YY=year of test (01) MM=month of test (07) DD=day of test (12) a=first sample list (run) of the day (the next sample list will end with `b,' the next `c' and so on.) 12.1.2 Assign a filename using the instrument designator letter, the last 2 digits of year-moday, and a 3-digit sequential file number that starts with 1:md increases by one for each filename. Example File Name: IYYMMDD### or DO10712001 I=instrument name (D for "Davey") YY=year of test (01) MM=month of test (07) DD=day of test (12) ###=3-digit sequential file number starting with 1through 999 (001) Also, as part of the sample list, assign a method (MS) for acquiring, an inlet file, a bottle number, an injection volume and sample descriptions. 12.1.3 To create a method, click on Method Editor button in the MS Status Pane and select SIR (Single Ion Recording) or MRM (Multiple Reaction Monitoring). Set Ionization Mode as appropriate and mass to 499 or other appropriate masses. Also set the acquisition start and stop times. Save acquisition method. If MS/MS instruments are employed, additional product ion fragmentation information may be collected. See Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information and MRM. 12.1.4 Typically the analytical batch run sequence begins with solvent blanks and a set of extracted matrix standards. 12.1.5 Sample extracts are analyzed with two CCVs injected every one to ten samples. Solvent blanks should be analyzed periodically to monitor possible analyte carryover and are not considered sample extracts but may be included as such. 3M Environmental Laboratory' ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 5 of 11 001046 Page 50 3M Medical Department Study: T-6295.25 418-018-.PAGE 1-424 Analytical Report: FACT-TOX-169 LRN-E01-0180 12.2 Using the HPLC 12.2.1 Set up sample tray according to the sample list prepared in Section 12.1.1. 12.2.2 Set-up the HPLC to the following conditions or at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook: 12.2.2.1 Sample size = 10 pL injection 12.2.2.2 Inject/sample = 1 12.2.2.3 Cycle time = 10.0 minutes 12.2.2.4 Flow rate = 300 pL/min 12.2.2.5 Mobile Phase (program) Time 0.00 min. 1.00 min. 5.50 min. 7.50 min. 8.00 min. MeOH 10% 10% 95% 95% 10% 2.0 mM Ammonium acetate (in H20) 90% 90% 5% 5% 90% 12.3 Instrument Set-up 12.3.1 Refer to ETS-9-24 for more details. 12.3.2 Check the solvent level in reservoirs and refill if necessary. 12.3.3 Check the stainless steel capillary at the end of the probe. Use an eyepiece to check the tip. The tip should be flat with no jagged edges. If the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary. The probe should be checked weekly. 12.3.4 Set HPLC pump to "On". Set the flow to 10 - 500 pL/min or as appropriate. Observe droplets coming out of the tip of the probe. 12.3.5 Turn on the nitrogen. A fine mist should be expelled with no nitrogen leaking around the tip of the probe. Readjust the tip of the probe if no mist is observed. 12.3.6 The instrument uses these parameters at the following settings. These settings may change in order to optimize the response: 12.3.6.1 Drying gas 250-400 liters/hour 12.3.6.2 ESI nebulizing gas 10-15 liters/hour 12.3.6.3 HPLC constant flow mode, flow rate 10 - 500 pL/min 12.3.6.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the HPLC is operating correctly.) 12.3.7 Carefully guide the probe into the opening. Insert probe until it will not go any further. Connect the voltage cables to the probe. 3M Environmental Laboratory ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 6 of 11 001047 Page 51 3M Medical Department Study: T-6295.25 418-018:PAGE 1-425 Analytical Report: FACT-TOX-169 LRN-E01-0180 12.3.8 Print the tune page, MS file, HPLC parameters, sample list and the Microsoft Word summary page and store in the study binder with a copy taped into the instrument log. 12.3.9 Click on start button in the MassLynx main page (this may vary among MassLynx versions, see appropriate MassLynx USER'S GUIDE). Ensure start and end sample number includes all samples to be analyzed. 13.0 Data Analysis and Calculations__________________ ______________________ 13.1 Calculations: 13.1.1 Calculate matrix spike percent recoveries using the following equation: __ % Recovery = -O--b-s-e-r-v-e-d--R--e-s-u-l-t-----M--a-t-r-i-x-B--la--n-k--R-e--s-u-l-t x 100 Spiking Level 13.1.2 Calculate percent difference using the following equation: %,, D,,,if,,ference = E--xp-e-c-t-e-d--C-o--n-e-.-----C--a-l-c-u-l-a-t-e-d--C-o-n--e-. x 10(1 Expected Cone. 13.1.4 Calculate actual concentration of PFOS, or other fluorochemical, in matrix (flg/mL): ( Cone, o f PFOS Calc, from Std. Curve {n g / m L ) x Dilution Factor ) {( In itia l V olum e o f M a trix (m L ) + m L o f S u rro g a te S ta n d a rd ) | Final Volume ( mL ) J 1gg 1OOO ng 14.0 Method Performance___________________________________________________ 14.1 Method Detection Limit (MDL) and Limit of Quantitation (LOQ) are method, analyte, and matrix specific. Please see ETS-8-4.2, Attachment B, for a listing of current validated MDL and LOQ values. 14.2 Solvent Blanks, Method Blanks, and Matrix Blanks 14.2.1 Solvent blanks, method blanks, and matrix blanks values must be below the lowest active standard in the calibration curve. 14.3 Calibration Curves 14.3.1 The coefficient of determination value for the calibration curve must be 0.990 or better. 3\ i Environmental Laboratory ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 7 of 11 00104 Page 52 3M Medical Department Study: T-6295.25 418-018:PAGE 1-426 Analytical Report: FACT-TOX-169 LRN-E01-0180 14.3.2 All active calibration curve points must be within 25% of the theoretical value with the exception of the LOQ point, which may deviate up to 30%. 14.3.3 Calibration standards with peak areas less than two times the curve matrix blank must be deactivated to disqualify a data range that may be affected by background levels of the analyte. 14.3.4 Low or high curve points may be deactivated to optimize a linear range appropriate to the data. 14.3.5 Not including low or high points dropped to optimize the linear range, curve points may be deactivated if they deviate more than 25% from the theoretical value when the curve is evaluated over a linear range appropriate to the data. 14.3.6 One point below the LOQ may remain active even if it deviates more than 30% or has a peak area less than two times the matrix blank; however, the LOQ will be defined at the lowest point with acceptable deviation. 14.3.7 A valid calibration curve must contain at least 5 active points above and including the LOQ. 14.4 Matrix Spikes 14.4.1 The average matrix spike percent recoveries should be within 30% of the spiked concentration. Recoveries outside of this range should be discussed in the report. 14.5 Continuing Calibration Verifications (CCVs) 14.5.1 Continuing calibration samples within the linear range of the run must show a percent recovery within 25% of the spiked concentration. If a CCV is outside of this recovery, subsequent data should not be accepted. Acceptable data must be bracketed by the curve and passing CCVs. 14.6 If criteria listed in this method performance section isn't met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the appropriate logbook. 14.7 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report. 15.0 Pollution Prevention and Waste Management____________________________ 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass pipette waste is disposed in broken glass containers located in the laboratory. 16.0 Records________________________________________________________________ 16.1 The first page of each data packet generated for a study must have the following information included either in the header, in the footer or hand written on the page: study or project number, instrument, sample matrix and time point, date, and analyst. 16.2 A data packet includes the following: data review summary form, MassLynx quantify compound summary report for each target analyte, quantify calibration report for each target analyte (curve), method report, Word document listing set-up parameters, tune 3M Environmental Laboratori ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 8 of 11 001049 Page 53 3M Medical Department Study: T-6295.25 418-018:PAGE 1-427 Analytical Report: FACT-TOX-169 LRN-E01-0180 method report, MassLynx scanning method report, HPLC method report, sample list, and quantity sample report (chromatogram). 16.3 For each analysis, after printing the tune method report, Word document listing, set-up parameters, MassLynx scanning method report, and sample list, copy and tape into the instrument runlog. The original is maintained in the data packet. 16.4 On each page of the quantify compound report, quantify calibration report (curve), and quantify sample report (chromatogram), the following information must be included either in the header, the footer, or hand written on the page: study or project number, instrument, method, calibration (the method and calibration are usually assigned the same name), analyst, and date. 16.5 The analyst must date and initial the first page in a packet as long as their initials and date are electronically included on each page. If initials and date are no; electronically included, they must date and initial each page. 16.6 Summarize data using suitable software (e.g., Excel) for inclusion in the final report. See Attachment A for an example of a summary spreadsheet. 16.7 Back up electronic data to appropriate medium. Record the file name and location of backup electronic data in instrument log book. 17.0 Tables. Diagrams. Flowcharts, and Validation Data______________________ 17.1 Attachment A: Data summary spreadsheet. 18.0 References______________________________________________________________ 18.1 FACT-M-4.1, "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry 18.2 ETS-9-24.0, "Operation and Maintenance of the Micromass Atmospheric Pressure Ionization/Mass Spectrometer Quattro II triple quadrupole Systems" 18.3 The validation report associated with this method is ETS-8-4.0 & 5.0-V-l. 19.0 Affected Documents_____________________________________________________ 19.1 ETS-8-4.2, "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry" 3.\( Environmental Laboratori' ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 9 of 11 001050 Page 54 3M Medical Department Study: T-6295.25 418-018`.PAGE 1-428 Analytical Report: FACT-TOX-169 LRN-E0I-0180 20.0 Revisions________________________________________________________ Revision Number. 1 Reason For Revision Section 6.1.2 Clarification of HP1100 system components. Section 11.1 Average of two curves, not standard values, are used for plotting linear regression and added the 1/x weighting of the: curve. Section 12.2.2.4 Clarification of solvent ramp. Section 17.1 Changed from attachment B to A. Revision Date 04/02/99 2 10.1 Clarified when blanks are run. 10.2 Added instructions for when surrogate matrix is used. 10.3 Specify requirements for CCVs. 11.1 Requires only a calibration curve before the samples. 11.2 Clarify what to do if curve does not meet requirements. 11.3 Allow to truncate the curve. 12.1.1 Clarify sample list ID. 12.1.3 Clarify typical run. 12.2 Changes to mobile phase gradient and specify flow rate. 14.3 Change acceptable limits. Add specifics for acceptance of calibration curve. 14.3.3 When to deactivate calibration standards based on blank response. 14.3.5 When to deactivate calibration standards within the linear range. 14.4 Modifies evaluation and use of matrix spikes. 14.5 Describes evaluation and use of CCVs. 14.5.1 Evaluation of CCVs. 16.0 Add requirements for records and documentation. Attachment A: Summary Spreadsheet ETS-8-5.2 Analysis of Serum Extract Using ES/MS 3M Environmental Laboratori' Page 10 of 11 001051 Page 55 3M Medical Department Study: T-6295.25 418-018 .-PAGE 1-429 Analytical Report: FACT-TOX-169 LRN-E01-0180 Laboratory Study # ' Study: Test Material: Matrix/Final Solvent: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Filename: R-Squared Value: Slope: Y Intercept: Date of Extraction/Analyst: Date of Analysis/Analyst: Group Dose Sample# Concentration ue/mL Initiai Vol. Dilution Factor Final Cone. ue/mL Slone: Taken from linear reercssion equation. Groun/Dose: Taken from the studv folder. Sample#: Taken from the studv folder. Concentration (uu/mLl: Taken from the MassLvnx integration summary. Initial Volume (mL): Taken from the studv folder. Dilution Factor: Taken from the studv folder. Final Cone. Cue/mL): Calculated bv dividing the initial volume from the concentration Attachment A: Summary Spreadsheet ETS-8-5.2 Analysis of Serum Extract Using ES/MS 3.\f Environmental Laboratory Page 11 of 11 001052 Page 56 418-018:PAGE 1-430 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Appendix D: Data Summary Tables Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Table 7. T O X 169 Data Summary of PFOS Concentration--Rat Serum (jjg/mL) Tlm epolnt Fem ale F0 Gestation Day 21 Pups F I Gestation Day 21 Female F0 Lactation Day 5 Pups F I Lactation Day 5 Group Group 1 0.5% Tween 80 0.00706 0.00280 (n=6) <LOQ* (n=6) 0.0184 0.00869 (n=6) 0.0299 0.0198 (n=6) Group 2 mevalonic acid 0.0183 0.0234 <n=6> 0.0782 0.0836 (n=6) 12.7 27.4 (n=6) 3.33 3.14 (n=6) Group 3 1 .6 mg/kg PFOS + mevalonic acid 55.8 8.87 (n=6) 207 25.6 (n=6) 127 24.7 (n=6) 12418.7 (n=3) Group 4 2 mg/kg PFOS + mevalonic acid 82.4 22.6 (n=6) 251 55.9 (n=6) 189 482 (n=6) N/A Group 5 cholesterol 0.0251 0.00396 <n=6) 0.0828 0.0315 (n=6) 0.688 1.62 (n=S) 0.1880.199 (1=6) Group 6 1.6 mg/kg PFOS + cholesterol 79.9 155 (n=6) 154 322 (n=6) 104 14.6 (n=6) 109 7.85 (n=2) Group 7 2 mg/kg PFOS + cholesterol 371 214 <n=6) 166 50.4 (n=6) 145 7.59 (n=6) 155 (n=1) Group 8 0.4 mg/kg PFOS N/A N/A 272 18.7 362 3.84 (n=6) (n=6) Group 9 0.8 mg/kg PFOS G roup 10 1 mg/kg PFOS N/A N /A N/A 426 6.92 53.1 27.7 (n=6) (n=6) N /A 52.3 * 2 6 ,0 84.4 17.5 (n=6) (n=6) Group 11 1.2 mg/kg PFOS N/A N/A 86.0102 147 162 (n=6) (n=*6) Group 12 1.6 mg/kg PFOS 142 24.5 (n=6) 142 332 (n=6) 169 32.1 (n=6) N/A Group 13 2 mg/kg PFOS 125 19.8 (n6) 170 38.1 (n -6 ) 134 26.6 (n=6) 138 (n=1) ` LO O -- Lim it at Q uantitation = 0 .0 0 4 9 4 pg/m L N /A-- Not applicable N O TE : Results are expressed as group averages the standard deviation associated with the group. N O TE : It is not possible to verify true recovery of endogenous analyte front tissues without ra d o-labated reference m aterial. T h e only m easurem ent of accuracy available at this tim e, extracted calibration curves and m ulti-level continuing calibration verifications (C C V s ), in dicate that the sera data are quantitative to 25% . 3 \f Environmental Laboratory- 001053 Page 57 3M Medical Department Study: T-6295.25 3M Medical Departm ent Study: T -6295.25 Appendix E: Data Spreadsheets 418-018:PAGE 1-431 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FA C T T O X -1 69 L R N -E 0 1 -0 1 8 0 3.U E n viro n m en ta l L aboratory' 001054 PageM 418-018:PAGE 1-432 3M Medical Department Study: T-6295.25 FACT-TOX-169 Analytical Report: FACT-TOX-169 Argus 418*018 LRN-E01-0180 Study: Eni-0180 FACT-TOX-169. Argus 418-018. One Generation Reproducaoa Study of PFOS-Mevalaaic AcxVChoklierai Challenge end NOEL Investigation in Riu Product NumbeiiTeii Subitane*): NA Muni: Rii Seri Method/Revision: ETS-MJ4ETS-8-5T! Analytical Equipment Synem Number Soup 020199, Amelia062498 IftstruRient Sofrwaie/Version: Maasiynx 3.4 Blenime: See Below R-Squared Value: See Attachmenls Slope: Y-lniercepc Dates of Extraction/Analyst: Dates of Analysis/Analyst Date of Data Reduction/Analyst See Attachmeau See Auachmenu 02/1M)l-02/20/01 RWW/KJS 02/27/01.03/ISA)l, 03/26/01 MMH 02/28/01.03/164)1, Q3/27AH MMH Sample Data Boa 40I-005A-E R AT SERA F0 GD21 Group Dom Group 1 0.31 Tween K0Control Group 2 Mevalonic Acid Control Group 3 1.6 mg/kg PFOS + Mevalonic Acid Group 4 2 mg/kg PFOS * Mevalonic Acid Group S Cholesterol Control Group 16 mg/kg PFOS Cholestemi Group 7 2 mg/kg PFOS Cholesterol Group U 1.6 mg/kg PFOS Group 13 2 mg/kg PFOS Sample 4 10901F 109OEF 10903F lOXUF* I0905F* I0907F 109IF 109I6F* 10917F I09I8F I0919F 10920F 1U930F* 10931F 10932F 10933F 10934F 1935F* I945F 10M6F I947F* Ujy49F 10930F 109IF UNS8F 10959F 10960F* 10961F* I962F* 10963F* 10972F* 10974F* 1097JF 10976F 10978F 109B0F I0983F 10987F* I09KAF 109WF 10990F 1099IP U040F" II044F11045F* 11046F 1HM7F II050F USJF I1054F I1056F U058F I1060F 11U62F* Initiai Vol. mL 0.70 0.50 0.71 0.43 0.43 0.73 0.6 0.48 0.35 0.73 033 060 0.30 0.J0 0.55 a63 0.69 0.36 0.70 071 a io 0.61 065 0.55 05ft 0250 0.37 0.29 0.43 047 0.45 0.38 053 0.55 0.52 072 0.71 0.20 064 051 063 11*7 0.39 043 0.40 0.65 056 0 63 0.64 0.81 0.70 0.73 058 0.43 TOPFOS Verified NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Confirmed Low NA Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low NA NA NA NA NA NA Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Law Confirmed Low Confirmed Low Confirmed Low Confirmed Low NA NA NA Confirmed Low Confirmed Low Confirmed Law Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low PFOS DUuttoo Fader l J 1 1 l 1 1 1 1 1 I 1 400 400 400 400 400 400 1000 1000 1000 1000 1000 1000 1 1 l 1 l l 1000 1000 IU00 1000 toon in m 1000 1000 1000 30) sorai iono inxi 1000 tono term 1000 10) 1000 mno 1000 1000 1 1000 PFOS Cene. ng/mL 836 no 603 2.25 3.73 5.26 39.3 3.97 6.24 4.45 4.31 6.34 33.0 69.3 75.0 69.0 88.9 52.9 62.9 38.2 7.15 73.9 48J 46.2 19.2 11.8 9.16 6.59 9.7R 11.2 44.4 24.3 51.9 43.8 38.6 467 71.6 87.1 78.8 64.6 64.6 9 6.2 TOO 64.7 61.5 71J 75J 78.7 74.2 92.0 77.8 84.3 77.8 69J Filename SOI0227020 S0102Z7021 SO10227022 S010227023 S01Q227024 SO10227025 SO10227029 S010227Q30 5010227031 S01Q227002 SO10227033 S010227034 A010315Q20 A0IO315Q2I A0I0315022 A010315023 A01Q315CQ4 AD10315025 A10315029 A010315000 A01Q315031 AL0315032 A01Q315033 A010315034 S010227035 SO10227036 SO10227037 SO10227038 A01Q315018 A010315019 AO10315035 Am03lS036 A010315037 A010315Q38 A01Q315042 A010315043 A010315044 A010315045 AO10315046 A01Q326016 A010326017 A 01032M I1J AGUE15030 A0I03I5Q3I AD10315055 AO10315036 A0I0315G57 A010315Q38 A01Q315QS9 AOIO31SQ60 AA10315061 A0I0315062 A010315063 A010315064 ConcewratMM <tfPlt)S ug/mL or % Rae 0.0119 <LOQ (0.00194 ug/mL) 0.00*38 <LOQ (0.00194 ug/mL) <LOQ (0.00194 ug/mL) 0.00/21 0.0/35 0.0124 00113 <LOQ (0.00194 ug/mL) <LOQ (0.00194 ug/mL) 0.0106 707 55 545 438 51 58 7 89R 53 9 715 121 74 3 83 9 11.0:30 0.K35 0.0148 U.K27 0.0:27 0.0138 98 7 639 98 0 796 742 648 101 436 123 633 5t3 41U J79 131) 154 nu 135 125 116 114 111 112 134 161 Mean pros ug/mL 000706 0.0183 55.R 82.4 0.0251 79.9 371 142 125 uso StcL Dev. MS/MSD RPD 396 0.00280 128 00234 15.9 8.87 27.4 216 15.8 0.00396 19.4 13.5 7.R 214 17.2 24.5 15.9 198 PFOS Pcrftuoforctineiulionaie * Ealtmiied concentration*, minai volume below0.5 mL. LAC 02/22/01 Date Emerod/By: Due Verified/ By: 03AH/D1.03/22/01 LAC/03/284)1 UH 04/06/01 hoi 3M Environmental Laboratorio 001055 Page 59 418-018:PAGE 1-433 3M Medical Department Study: T-6295.25 FACT-TOX-169 Analytical Report: FACT-TOX-169 Argus 418-018 LRN-E01-0180 Study: 014)180 FACT-TOX-169, Arius 418-018. One Generation RepnxUxoon Study of PFGS-Mcvakuuc Aad/Cbefcaterol Chalknje and NOEL Invesdcaikm in Rjos Product NurebeifTeal Substance): NA Matrix: Rat Sera Meiitnd/Revuion: ETS4W.2 A ETS-8-5.2 Analytical Equipment System Number Soup 020199, Amelia 062498 Instrument Softwane/Venion: Masslynx 3.4 Filename: Sae Below R-5qlured Value: SWipe: See Attachments See Attachmeats Y-lntercept See Attachments Dates of Extraction/Analyst: Dates of Analysis/Analyst 02/13/01-02/204)1 RWW/KJS 02/21/01.03/2001, MMH Dale of Data Reduction/Analyst 0311,03/21/01.3/2SWI MMH Sample Data Box #01-005A-E R A T SERA PUPS F l CD21 Group Date Group 1 0_5%Tween 80 Cimimi Croup 2 Mevaimic Acid Conimi Group 3 l.6mg/k| PFOS * Mevalonic Acid Group 4 2 mg/k| PFOS -*Mevalonic Acid Group S Cholesterol Control Sample # 10901P10902P* 109Q3P* 10904P* 10905P* I0907P* 915P* ID916P* 10917P* 10918P* 10919P* 10920P* 10930P* 10931P* KW32P* 10933P* 10934P* 10935?* 10945P* 10946P* 10947P" ID949P* v10m95\0?P** 10y58P* 10959P* 10960P* I0961P- Initial Voi. mL 0.14 ami 0.13 0.05 0.18 no6 0.24 o. m 0 1* 0.06 0.09 0.03 0.19 0.23 0.22 0.15 ai9 0.21 0.18 0.16 0.13 0.07 0J0 0.23 0.10 0JI 0.11 0.28 THPFOS Verified NA NA NA NA NA NA NA NA NA NA NA NA NA NA 2nd analysis OK 2nd analysis OK NA NA 2nd analysa OK Confirmed Low Confirmed Low Confumed Low 2nd analysis OK 2nd analysis OK NA NA NA NA PFOS Dilution Factor 1 1 1 1 1 1 1 1 1 1 40) 400 400 400 400 400 400 4(1) 400 400 400 4on i i i i PFOS *C%o/nmeL. 2.96 (1470 4.20 o.no 3.75 1.77 40.4 1.30 19.6 3.62 15.9 non IU2 99.1 120 90.7 102 954 119 905 83.3 60.4 131 137 14.0 1M 9.88 20.1 Filename SAI0228112 9010228113 S0I0228 U4 901022*115 SO10228 lift SD10228II7 S010228I2I 9010228122 9010228123 9010228124 SO10228125 901022*126 A010320016 A10320017 A01328016 A0I0328017 AOIO32U20 A010320021 601032*018 6010320023 Am0320n24 Amo32oai3 A0I032*019 A010328020 501022*127 SO1022812* SOIG228129 S0I0228130 Coueamtresiu of PFOS U|FmL or 7 Ree <LOQ <0.00491 ug/mLl <LOQ (0.00*91 uj/raL) <LOQ (0.00491 og/reL) <LOQ (0.00491 ug/mL) <LOQ (0.00*91 ug/raU <LOO (0.00491 uiMtL) 0.16* <LOQ (0.00491 ug/mLl 0.109 <LOQ (0.00491 u|/mL) 0.177 <LOQ (0.05491 urAnL) 214 172 219 242 214 1*2 265 226 256 345 175 239 0.140 0.0341' Q.089V ao74: Mean PFOS UR/mL. RSD SkL Dev. MS/MSD RPO <LOO 0.0782 NA NA 107 0.0*36 12J7 207 23.6 22J 251 35.9 Group 6 1.6 m*/k* PFOS Cholesterol Group 7 2 mc/kf PFOS Cholesterol 10962P* 0963P- I972P* 10974P* 1097SP* 10976P* 10978P* I0980P* 1098SP* 10987P* I09**P* 10989P* lOWOP* 1099 IP* 0.28 0.31 0.13 026 0.09 0.16 0.16 0.07 0.24 0.18 0.21 0.06 0 14 024 NA NA Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low NA Confirmed Low Confirmed Low Confirmed Low i 23.1 S10228I34 an*2/. i 17.1 S01022813S 0.055:- 400 43-S A0I0320Q31 140 400 85.2 A010320Q32 131 400 Mi) AU10320(03 196 400 575 A010320Q34 145 400 4*3 AJ)10320003 121 400 33.9 AU10320036 194 400 73-3 601(021)037 126 400 67J Am0320n3S 150 400 *5.1 6010320042 162 400 39.6 A010320043 264 4tn 35.4 AO10320044 151 400 80 2 AUKl32nvtS 134 0.0818 154 166 3*.fl 0.0315 20.9 32.2 304 304 Group 12 1.6mg/kj PFOS Group 13 2mg/kf PFOS nwop* U044P* 11045P* IM46P* 11047P* 11050P* l1053P* 11054P* U0S6P* U158P* U060P* 1I062P* D8 0.18 0.15 0.25 0.11 0.21 0.11 0.03 008 0.29 0.17 0 20 Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low Confirmed Low NA Confirmed Low Confirmed Low Confirmed Low 4U> 39.2 A01032fXM6 1% 400 57.1 Afi)0320047 127 400 47.7 AO1(132004* 177 400 69.8 A010320049 112 400 46.4 Afll(020050 169 400 61* AOIO320Q51 120 44X1 41.7 6010320055 152 400 28.4 AI0320056 227 400 39.4 6010320057 197 400 913 ADKOMQ5* 126 400 593 A01Q320QS9 140 400 887 AD10320060 177 23.4 142 33.2 22.4 170 3*. 1 PFOS Perflunmnciancsuifonaie * Estimated concentration*, initial volume below0.5 ml--LAC02/22/QI Date finiered/By: Date VenTied/ By: 0307/01. I3/23I LAC/04/03/01 IH 04/06/01 ho) 3.1/ Environmental Laboratory 001056 Page 60 418-018-.PAGE 1-434 3M Medical Department Study: T-6295.25 FACT-TOX-l Analytical Report: Siudy: EOl-OiaO Pmdect Nurtm-fTeUSubeLancel. Argus 418-018 FACT-TOX-169. Affai 41(4)11.OnaG an u m Riymrhiwn Stpyni PTOS>Mev*tate AnhirtriananW Okaltef t e NOELti NA team ed SoftwanWar faiSm ET3-I-4.2 * m - l- J J Atei* 061491. S<mp020199 M teyB 3.4 SeeBdow R-Sqvnd Vate: Slope: FACT-TOX-169 LRN-E01-0180 DatoUEnraoteAMly: Duca o Aaaiysii/AteysC Due o EUJUduoteAjulysL- 02/13-01-02/2001 RWW/KJ5 031/OI.0V1MD1. OSAMI. OSAMI.03/3001 MMN 0WI2I. 0VIMD1.0V27l.0V9I, W2I MMH Sample Data Boa BOI-0QSA-E RAT SERA FOLDS_______________ _____ Gnep tkm (imp 1 0.3*Tween SOOverol Gmaf 2 Mcvafcrtc Add C i t e Crampi 1.6*fefPF0S * Mcnlranc Aed Crampi lraytiPFOS f Mvalerte And Grana 3 Q rtcam t Craeml Grampi (Arnesi PFOS Onleilte Gnraa 7 2mpfc|PFOS Orteaieml (roup 1 0.4 p ro s Crimp 9 a a maAaPFos Cramp 10 i maAaPros (ramp 11 1.2mgAt PTOS Cramp 12 1.6m pt| PFOS (ramp U 2mAgPP0S Sample 1W06F* 1Q9Q9F 1091IF' 11301F* I13GQF 11303F* 10921F 10922F* 10913P* 11513F" listar* 115171* 10929F* I093BF 10942F* IIS36F IIS37F* 11S38F* 11544F 11545F 11544F* 1JJ47F* 11549F 11S50F* 10966F* I0IM7P* 10968F* 11531F 11339F 11361F* IOV77F 10979F 10913F* 1157F* 11373F 11574F 10995F10997F* tisaaF 11517F* U5UFM5R9F 1CMWF 11000F* 11003? 1I60F* 11602F 11603F* 11009F 11012F noijp11612F 11614F 11617F 1101VF* 11021F* 11023F* U621F* U624F* 11623F* 11031F* 11032F* 11033F Il630F 11634F* 11639F* 110*1F 11049F* 11640F* 11641F* 11642F 11643F 11039F 11634F 11633F* 11636F* U6STF* I165BF Irate VU. mL 0.36 062 0.39 0.43 0.32 0.36 0J3 0.44 043 032 0.13 0.23 0.44 OJO 0 3 0.65 0.11 0.41 0JB 100 044 010 0.66 0.21 0*9 0.14 0.43 0-65 0J0 0.43 0J3 QJ3 031 QJ7 0.33 0.60 040 0.40 0.64 0.46 Q.40 1.00 0J3 a47 0J3 029 0.63 0.37 0.64 0J3 0. Q.M 071 1163 0.41 0 3 a33 0J3 0.4 0.43 0.31 0J7 0J3 0J2 Q2i 0.13 0J9 0J2 an 0.12 1.00 0J6 0J1 0J2 0J1 aii ou 072 Turros V teU NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 2nd t e t e OK NA NA NA NA t e analy* OK NA NA NA NA NA CarBtnte Hifh NA NA NA NA 2ndteyaii OK NA NA NA NA NA NA NA NA NA 2ad t e t e OK t e t e t e OK t e U te ok 2nd a t e t e OK t e t e t e OK teanahakOK 2nd ateyiu OK OnAiraradLo 2M ateyau OK t e t e t e OK t e t e t e ok 2nd t e t e OK 2nd a t e t e OK t e rauBytiaOK t e t e t e OK te t e t e ok NA 2ndtev>uOK 2nd ateyi OK 2ndt e t e OK t e t e t e OK t e te y ra OK t e t e t e OK t e t e t e OK te te y m i OK t e t e t e OK t e te i u 0K t e t e t e ok t e t e t e 0K t e t e t e OK t e t e t e ok t e t e t e OK t e t e t e ok t e t e t e ok t e t e t e OK t e mlraii OK rros radar I 1 1 t l 1 1 500 1 20 20 20 1230 1230 1230 1230 230 1230 1230 2300 1230 230 2500 1230 1 1 V 30 1 1 1230 1230 230 liso 1230 1230 1230 1230 1230 1230 1230 1230 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 tooo 1000 1000 1000 1000 90 1000 1000 1000 1000 1000 1000 1000 1000 1000 (000 1000 1000 1000 2000 2000 2000 3000 2000 2000 r r o s FUrtarara Orne. 7.4B SOI0301020 13.7 S01030102I 4.42 SOI0301022 140 5010301023 IJ3 S0IO3Q1O24 3.23 SO10301023 27.4 AOI31AOI8 60.2 A010326OIV 6.66 A010316019 44.3 A0IO3I6O29 13.1 A0I0116O30 27.3 AOI0316031 36.2 A01326020 64.9 A010326021 31.7 A010324022 69.6 AQ10326023 (2.6 A0I032S0Z2 36.1 A010326024 60.1 A0I0326025 79.3 AOI0326029 3I.B A0I0326030 1B A010328023 42.5 A010326031 34.8 A010326032 11.2 A0I03IAO20 1.71 A0I0316O21 ia A010316022 31.1 A0I0316O47 29.6 A0IO316O23 10.1 A0103I6024 49.5 A010326033 33-0 A0IO326034 124 A010328023 35.7 A010326035 49.8 A010126036 48.0 A010326017 46.0 AG10326031 43.2 A010326042 (0.3 A0IO326O43 54 1 A01032644 44.4 A01032604S 118 A01O326O46 9.61 A01Q328029 7.89 A010328030 10.4 A01032B03I 18.1 A0I032I032 16.3 A01032U33 7.31 AQI0328Q14 23.3 A010328035 21.2 AOI03I6073 Z3.4 A0103280JT 36.0 AD1032S031 4<L2 AOI0330017 21.6 A01033001B 21.1 A0I330019 19.8 A0I0330020 26.7 A0I0330021 20.3 A0I0330022 16.1 AOumao7 28.7 A0I0330023 340 A0IO330024 26.2 A010330023 40.4 A010330026 4(6 A010330030 24.9 A010330031 9.20 A010330011 12.4 A01Q33003) 613 A010330034 2a i A010330033 24 1 AOI0330034 120 AOI0330037 985 A0I0330038 37.7 A010330039 412 AOI0330043 14 3 ADIOS30044 211 A010330045 m A01033004A 537 AOI0330O4T f FOS Pdlterncuraetenraw * E te te d onpoMUUicna. l te te w teP 0 SwL LACQS/H! Caracaralrvdara w rros at/m lar 18 Pt -0208 00220 <LOQ(000494 atte-) acni aoi7o 0.0146 0.0320 68.5 0.0133 271 142 138 103 162 102 134 115 146 131 199 167 270 161 207 0.0228 0.0122 0.0253 3.99 0.0592 0 0223 112 79.6 100 i21 113 too 144 135 157 147 139 147 17.5 16.8 1(9 649 23.1 19.8 34.3 40.0 4 4 .1 43.0 51.3 34 2 38.5 310 76.2 61.5 IW 637 89.4 97.0 76.3 93.3 88.9 70.1 140 195 IR 201 120 176 142 162 913 III 141 149 Moan rros uteL 0.0184 12.7 127 189 0688 104 143 ru 42.6 32.3 16.0 169 134 BSD SU. Dvt. MS/MSDRTD 47.2 0.00849 216 274 19.5 24.7 25.5 48.2 235 t.62 140 14.6 3.24 7.39 690 1*7 16.2 6.92 498 26.0 11.9 10. 190 321 199 26.6 DaaeEtemVBr Daae Vanite I r OSAMI.03/Z2/01 LAC/OSAftOl.MOSAl KJH 040*01 Ini SM Environmental Laboratori 001057 Page 61 418-018:PAGE 1-435 3M Medical Department Study: T-6295.25 Analytical Report: FACT TOX-169 7 FACT-TOX-169 7 A rgus 418-018 LRN-EOI -0180 Study: E01-0180 FACT-TOX-169, Argus 411-O lt, One Generation Reproducson Study otPFOS-Mevalonic A d d Q o k c st:ro i Challenge and NOEL Investigation in Rats Product NumbofTesi Substance): NA Mania: Ru Sen Method/Revisioa: E T S -fM J i t CTS-8-5.2 Analytical Equipment System N um ber Soup 020199 Insuument Sofiware/Venion: M aislyn* 3.4 Filename: See Below R-Squared Value: See Attachaient! Slope: See Attachments Y-Ifiteicqn: See Atuchm eau Dates o f ExtracikmWAnalyse O2/13/01-Q2/2Q/D RWW/KJS Dates oi Analyiu/Analyst: 03WJ1.03/21I. 03/26AJ1. OV2AOI. (13/3001 MMH Date or Data Reduction/Analyst: 03/13/01,03/22/01.03/278)1,03/294)1.04/02)l MMH Sample Data Box 01-003A-E RAT SERA PUPS FI LD5 Group Dow Group 1 0 5 % Tween 80 Coatrol S anpto# 1U906F* 10909F* Initial Voi. mL .3 (1.04 Varillad NA 2nd Analysis OK PFOS Dilution F acte r 1 1 PFOS Coac. nf/m L no 0.00 FUro a m a S0103080I8 A01UQ210I9 C o a ca se ra tm of PFOS liW . ot % Rae 1.0366 <LOQ (C.00494 ug/fnL) M ean PFOS ug/m L RSD S ld. Dcv. MS/MSD RPD G roup! Mevalonic Acid Control I09U F * 11501/6F U302/7F 11503/8F 1092IF* I0922F* 10923F* ao8 0.95 050 0.64 0.20 0.31 0.06 NA NA NA NA NA NA 2nd Analysis OK 1 0 0 0 S01Q308020 <LOQ(T.00494 ug/raL> l 388 S010308021 ').0408 l 24.9 $010300022 >.0497 27.1 $010308023 .0423 0 0299 1 20.6 SO138024 a 103 l 6.62 9010308023 0.0214 l 1.78 A0I032I029 <LOQ (0.00494 ug/mL) 66.2 0.0198 II5I5/24F 1.00 NA 100 49.8 AOJ32136 498 U516/21F* 0.41 NA 100 30.5 AO1CO21037 7.09 94.4 I15I7/20F 0.76 NA 100 33.7 A010321038 4.43 3.33 3.14 Group 3 16 mg/kg PFOS * Mevalonic Acid IQ929F* 1094IF * 11536/37F* a io 0.05 a io NA NA NA 20 51.6 AOI032142 103 200 34.8 A010321043 139 200 64.7 AIQ371044 129 13.1 124 18.7 G roupS 10966F* 0.13 NA 1 2.96 A0L032130 <LOQ (0 00494 ug/mL) Cholesterol Control 10967F* 023 NA I 18.1 S0I0308034 0.0787 M969F* a io NA 1 7.84 S0I038035 0.0784 115JS/69F 066 NA 355 S01030S036 1538 U559/70F 1.00 NA 166 SO10308037 1166 106 11561/68F 1.00 NA l 80.5 S01030803 0805 0.188 (1.199 Group 4 I0979F* 0.11 NA 200 618 A01O32104S 114 7.22 1.6 mg/kg PFOS Choi. 10983F* 012 NA 200 61.8 4010321046 103 109 785 G ro u p 7 2 m ^ k g 9 -Choi. 1UW7F* 0.30 NA 500 93.3 A10321047 155 NA NA G roups I0999F' au NA 200 17.7 AO10326048 311 0.4 m g/kf PFOS IIOOOF- 0.23 NA 200 48.4 A010326049 42.1 110Q3F" 0.09 NA 200 17.3 A01Q326050 38.4 HOttF* 0.47 NA 200 75.4 AO1O321051 32.1 IIM 2/9F * 0.28 NA 200 501 A01032105 35.8 10.6 Group 9 I160V4F* 11009F* 0.17 006 NA NA 200 31.2 A0I032605I 36.7 200 18.9 A010326055 18.9 36.2 3.84 0.8 mg/kg PFOS 110I2F* 0.15 NA 20U 532 A0IO32IO58 70.9 1I0I3F* U(J7 NA 200 21.1 A01032A056 21.1 U612/13F 0-30 NA 4on 73.6 A01Q32106U 58.9 116J4/16F* 0.35 NA 400 518 A0IQ32I06I 60.4 52.1 11617/19F* 0.21 NA 400 46.3 A0I0321062 88.2 53.1 27.7 G roup 10 110I9F* 028 NA inoo 27.0 A0I0326057 96.3 1 mg/kg PFOS u raiF - 020 NA 1000 131) A01032/058 65.0 1123F* 026 NA 1000 22.9 AIQ326059 88.2 11621/22F* I1624/29F' 1I625/28P 034 0 14 OTO NA NA NA iaon 3 1 0 A010326O6O 91.0 ino 85.8 A010321070 61.3 toou 733 AOtraaanai IOS 207 *4.4 17.3 G ro u p 11 1I031F* 0.08 NA 100 382 A01Q330048 477 t 1.2 mg/kg PFOS 1I032F* UQ33F* 0.16 0.30 NA NA tono 1000 IOS A/U0326063 28.8 A 010326064 55.4 76.0 11036F* 015 NA IOOO 9.05 A010326068 13 I1037F* 11634/39F* 0.17 031 NA NA IOOO tono 15.7 A01Q326O69 273 AOHH26m> 715 48.8 m 147 162 C ro u p 13 2 meflsg PFOS I1QS9F* 0.09 NA 20) 613 A01U3260T7I 138 NA NA PFOS Periluonanctanesulfonaie * Estimated concentrations, m isal volume haiow 0.} mL. LAC D2/224J1 a 4 point curve u used few quaiuiia&ao. KIHO4/O3/01 Date Entemd/By: 03/22/01.03/23/Dl LA C / 03/21/01. 04O31 KJH Dam Verified/ By: 04/06/01 hoj 3M Environmental Laboratory 001058 Page 62 418-018-.PAGE 1-436 3M M edical Department Study: T -6295.25 3M Medical Department Study: T-6295.25 Appendix F: Example Calculations A nalytical Report: F A C T -T O X -169 LR N-EO l-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Formula Used for Sera Analyses in Study FACT TOX-169 A R (ng/m L) x D F x FV (mL) x 1.0 pg = R ep orted C o n cen tra tio n (p g/m L ) E V (mL) 1000 ng Calculation Used for Group 3, FO, GD21, Animal ID 10930F 5 3 .0 ng/m L x 4 0 0 x 1,0 mL x 1.0 pg 0.3 mL 1000 ng = 70.7 pg/mL A R -- Analytical result from M assLynx summary DF-- Dilution factor FV-- Final extract volum e (1 .0 mL unless otherw ise noted) EV-- Volum e o f sera extracted 5 \i Environmental Laboratori' 001059 Page 63 418-018:PAGE 1-437 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Appendix G: Interim Certificate of Analysis SM Environmental Laboratori oaiofio Page 64 418-018:PAGE 1-438 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 _ . . . . . . _ LRN-E01-0180 Centre Analytical Laboratories, Inc. ( 1 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Revision 1(9/7/00) Centre Analytical Laboratories COA Reference #: 023-018A 3M Product: PFOS, Lot 217 Reference#: SD-018 __________________ Purity: 86.9% ________ T est Name Specifications Purity' Result 86.9% Appearance White Crystalline Powder Conforms Identification NMR Positive Metals (ICP/MS) I. Calcium 1. 0.005 wt./wt% 2. Magnesium 2. 0.001 wt/wt.% 3. Sodium 3. 1.439 wt./wt.% 4. Potassium2 4. 6.849 wt/wt.% 5. Nickel 5. <0.001 wt/wt.% 6. Iron 6. 0.005 wt/wt.% 7. Manganese 7. <0.001 wt/wt.% Total % Impurity (NMR) 1.93 wt/wt.% Total % Impurity 8.41 wt/wt.% (LC/MS) < Total % Impurity None Detected (GC/MS) Related Compounds - POAA 0.33 wt/wt% Residual Solvents (TGA) None Detected Purity by DSC Not Applicable' Inorganic Anions (IC) 1. Chloride 1. <0.015 wt/wt.% 2. Fluoride 2. 0.59 w t/w t.% 3. Bromide 3. <0.040 wt/wt.% 4. Nitrate 4. <0.009 wt/wt.% 5. Nitrite 5. <0.006 wt/wt.% 6. Phosphate 6. <0.007 wt/wt.% 7. Sulfate4 7. 8.76 wt/wt.% Organic Acids 5 (IC) l. TFA l. <0.1 wt/wt% 2. PFPA 2. <0.1 wt/wt.% 3. HFBA 3. 0.10 wt/wt.% 4. NFPA 4. 0.28 wt/wt.% Elemental Analysis": 1. Carbon 1. Theoretical Value = 17.8% 1. 12.48 wt/wt.% 2. Hydrogen 2. Theoretical Value = 0% 2. 0.244 wt/wt.% 3. Nitrogen 3. Theoretical Value = 0% 3. 1.74 wt/wt.% 4, Sulfur 4. Theoretical Value = 5.95% 4. 8.84 wt/wt.% V. 5. Fluorine 5. Theoretical Value = 60% 1 5. 54.1 wt/wt.% COA023-018A 3M Environmental Laboratori' Page 1 o f3 001061 Page 65 418-018:PAGE 1-439 3M Medical Department Study: T-6295.25 A nalytical Report: FA C T-TO X -169 L R N -E 01-0180 Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 Phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS C entre A nalytical L aboratories C O A R eference #: 023-018A Date o f Last Analysis: 08/31/00 Expiration Date: 08/31/01 Storage Conditions: Frozen <-10C R e-assessm ent Date: 08/31/01 'Purity = 100% - (sum o f m etal im purities, 1.45% +L C /M S im purities, 8.41% +Inorganic Fluoride, 0.59% +N M R im purities, 1.93% +organic acid im purities, 0.38% +PO AA, 0.33% ) T otal im purity from all tests = 13.09% Purity = 100% - 13.09% = 86.9% 2P o ta ssiu m is ex p e c te d in th is salt form and is therefore n o t co n sid ered an im purity. 3P u rity b y D S C is g e n e r a lly n o t a p p lica b le to m aterials o f lo w purity. N o endotherm w as observed for this sam ple. 4S u lfu r in th e sa m p le ap p ears to b e co n v erted to SO4 and h e n c e d etected u sin g the inorganic anion m ethod conditions. T he anion result agrees w ell w ith the sulfur determ ination in the elem ental analysis, lending confidence to this interpretation. B ased o n the resu lts, th e SO4 is n o t con sid ered an im purity.56 5T F A HFBA NFPA PFPA Trifluoroacetic acid H eptafluorobutyric acid N onofluoropentanoic acid Pentafluoropropanoic acid 6T h eo retica l v a lu e c a lc u la tio n s b ased o n th e em p irical form ula, C*;F 17S OaTC* (M W = 5 3 8 ) This w ork w as conducted under E PA G ood Laboratory Practice Standards (40 C FR 160). COA023-018A 3 \ t Environmental Laboratori' Page 2 of 3 001062 Page 66 418-018:PAGE 1-440 3M Medical Department Study: T-6295.25 A nalytical Report: FA C T-TO X -169 LRN-E01-0180 Centre Analytical Laboratories, Inc, k 3048 Research Drive State College, PA 16801 A k Phone:(814)231-8032 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS C entre A nalytical L aboratories C O A R eference #: 023-018A L C /M S Purity Profile: Im purity C4 C5 C6 C7 T otal w t/w t % 1.22 1.33 4.72 1.14 8.41 N ote: T he C 4 and C6 values w ere calculated using the C4 and C6 standard calibration curves, respectively. The C5 value w as calculated using the average response factors from the C 4 and C6 standard curves. L ikew ise, the C7 value w as calculated u sing the average response factors from the C6 and C8 standard curves. I. Prepared By: jbrtvid S . B eU D ate Scientist, C entre v ^ ily tic a l Laboratories // R eview ed By: ft /J o h n Flaherty v D ate Laboratory M anager, Centre A nalytical Laboratories V. COA023-018A 3X Environmental Laboratory Page 3 o f3 001063 Page 67 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Appendix H: Report Signature Page 418-018 :PAGE 1-441 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Deanna J A u ^ k e r ^ . S . , StuyDirector ( / Date I Marvin T. Case, D.V.M., Ph.D., Sponsor Representative LX Date Kristen J. Hansen, Ph.D., Principal Analytical Investigator leoi Date William K. Reagen, Ph.D., Laboratory Manager <3 Date 3M Environmental Laboratom 001064 Page 68 418-018:PAGE 1-442 P F O S C oncentrations in R at L ivers Southern R esearch Institute Study A 344.1.4 001065 418-018:PAGE 1-443 M ethods W e first prepared co n tro l tissu e h o m o g e n a te b y h o m o g e n iz in g rat liv er from con trol rats 1 :5 (b y w e ig h t) w ith D I w a ter. T h is w a s u se d to p rep are th e c a lib r a tio n stan d ard s. W e sp ik ed in k n o w n am ounts o f test article (P F O S ) and internal standard (P F O C ) into aliq u otes o f the b lan k tissu e h o m o g n ea te. A blank + int. std. and a b la n k - int. std w ere prepared w ith each calibration curve. T he sam p les w ere also h o m o g en ized (1:5) b y w eig h t w ith D I w ater. D ilu tio n s m ad e to any o f th ese sam p les w ere w ith u nspiked blank control liver h o m o g en a te prepared above. T hen an equal am ount o f internal stanard (P F O C ) w a s add to ea ch sam p le. T hen to both the sam p les and standards w e added 1 m L o f D I w ater, 1 m L o f 0 .2 5 /0 .2 5 M carbonate/bicarbonate buffer, and 500 pL o f ion pairing reagent (0.5 M tetrab u tylam m on iu m h y d ro x id e) adjusted to pH 10.5 w ith so d iu m h y d ro x id e. T he solution w as m ixed briefly and the w e added 2.5 m L o f ethyl acetate. T his m ixture w as shaken for 1 hr on a platform m ixer. A fter an hour the sam p les are taken o f f and centrifuged at 2 5 0 0 rpm for 5 m in. T he ethyl acetate layer is rem oved and evaporated to dryness under dry nitrogen. T he residue is reconstitued and filtered through a syringe filter into autosam pler vial for an alysis. W e then perform H PL C M S /M S usin g a M R M experim ent on the sam p les and standards. 001066 Southern Research Insitiute Study A344.1.4 P F O S in R at Livers File N am e S am p le N am e M easured C one. (ng/m L) 6/28/2004 6/28/2005 6/28/2006 6/28/2007 6/28/2008 6/28/2009 6/28/2010 6/28/2011 6/28/2012 6/28/2013 6/28/2014 6/28/2015 6/28/2017 6/28/2018 6/28/2019 6/28/2021 6/28/2022 6/28/2054 6/28/2024 6/28/2025 6/28/2026 6/28/2028 6/28/2029 6/28/2030 6/28/2032 6/28/2033 6/28/2034 6/28/2036 F0 10906 control F0 10909 control F0 10913 control F0 11501 control F0 11504 control F0 11505 control F1 10906 control F1 10909 control F1 10911 control F1 11501 control F1 11502 control F1 11503 control F1 10999 0.4 mg/kg F1 11000 0.4 mg/kg F1 11001 0.4 mg/kg F1 11600 0.4 mg/kg F1 11602 0.4 mg/kg F1 11601 0.4 mg/kg F1 11049 1.6 mg/kg F1 11640 1.6 mg/kg F1 11641 1.6 mg/kg F1 11642 1.6 mg/kg F1 11644 1.6 mg/kg F1 11648 1.6 mg/kg F1 11059 2 mg/kg F1 11654 2 mg/kg F1 11655 2 mg/kg F1 11656 2 mg/kg -5.582 1.553 -3.786 -0.2463 10.29 15.86 -9.268 -10.07 -8.904 -1.42 -10.5 -10.27 157.5 273.7 168.4 139.7 215 327.9 1245 541.7 601.9 539 592.7 650.1 660.3 632.2 665.2 604.6 6/28/2037 6/28/2038 6/28/2040 6/28/2041 6/28/2043 6/28/2044 6/28/2045 6/28/2047 6/28/2048 6/28/2050 6/28/2051 6/28/2052 6/28/2053 6/28/2055 6/28/2056 F0 11004 0.4 mg/kg F0 11600 0.4 mg/kg F0 11601 0.4 mg/kg F0 11603 0.4 mg/kg F0 11640 1.6 mg/kg F0 11643 1.6 mg/kg F0 11642 1.6 mg/kg F0 11645 1.6 mg/kg F0 11646 1.6 mg/kg F0 11659 2 mg/kg F0 11664 2 mg/kg F0 11657 2 mg/kg F0 11658 2 mg/kg F0 11059 2 mg/kg F0 11654 2 mg/kg 510.2 388.2 496.5 455 921 1342 1025 989.4 964.2 1389 1087 1196 1356 1080 1162 *Pup and Dam samples were mixed together during preparation BQL = below quantitation limit, < 12.5 ng/mL OR ng/g C alcu lated C one, (ng/g) BQL BQL BQL BQL 1,265 2,001 BQL BQL BQL BQL BQL BQL 29,745 96,373 101,752 43,318 71,429 98,027 116,800* 191,752 477,698 148,895 302,398 249,080 143,232 356,169 199,460 281,865 54,801 42,824 45,509 48,533 122,230 113,249 87,086 113,921 114,581 159,015 159,034 133,931 151,171 110,148 161,501 A PPEN D IX J H ISTO PA TH O LO G Y REPORT 001068 418-018:PAGE J-l RESEARCH PATHOLOGY SERVICES, INC. 438 East Butler Avenue, New Britain, PA 18901 Phone: 215-345-7070 Fax: 215-345-4326 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS PROTOCOL 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 HISTOPATHOLOGY REPORT SUBMITTED TO: Raymond G. York, Ph.D. , D.A.B.T. Associate Director of Research Argus Research Laboratories, Ine. 905 Sheehy Drive, Building A Horsham, PA 19044 SUBMITTED BY: * o - I?, W. Ray Brown, D.V.M., Ph.D. Veterinary Pathologist December 13, 2002 001069 418-018:PAGE J-2 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS PROTOCOL 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 HISTOPATHOLOGY REPORT TABLE OF CONTENTS Page REPORT Method.................................................................................................................................... 1 Results.............................................................................................................. 3 Summary.................................................................................................................................4 Quality Assurance Unit Statem ent..................................................................................... 5 001070 418-018:PAGE J-3 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS PROTOCOL 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 HISTOPATHOLOGY REPORT METHOD: Microscopic examination was made of the heart and thyroid of two male and two female F1 generation Crl:CD(SD) IGS BR VAF/Plus pups of a one-generation reproduction study. The dosage group identification and treatment of the groups of rats are listed below. Group 1 Identification Vehicle Control Number of Animals 28 1st Dosage RODI H20 2ni* Dosage '/ j hr + 10 minutes after 1st 0.5% Tween 80 3rd Dosage 5 hr + 30 minutes after 2nd RODI H20 2 Mevalonic Acid Control 3 1.6 mg/kg PFOS + Mevalonic Acid 4 2 mg/kg PFOS + Mevalonic Acid 5 Cholesterol Control 6 1.6 mg/kg PFOS + Cholesterol 7 2 mg/kg PFOS + Cholesterol 8 0.4 mg/kg PFOS 9 0.8 mg/kg PFOS 28 28 28 28 28 28 20 20 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/mg Cholesterol 500 mg/mg Cholesterol 500 mg/mg Cholesterol NA NA 0.5% Tween 80 1.6 mg/kg PFOS 2 mg/kg PFOS 0.5% Tween 80 1.6 mg/kg PFOS 2 mg/kg PFOS 0.4 mg/kg PFOS 0.8 mg/kg PFOS 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Add 500 mg/kg Mevalonic Add NA NA NA NA NA 10 1 mg/kg PFOS 20 NA 1 mg/kg PFOS NA 11 1.2 mg/kg PFOS 12 1.6 mg/kg PFOS 20 28 NA NA 1.2 mg/kg PFOS 1.6 mg/kg PFOS NA NA 13 2 mg/kg PFOS 28 NA 2 mg/kg PFOS NA The test and control articles were considered 100% pure for the purpose of dosage calculations. Mevalonic acid was dosed twice daily. The F0 generation female rats were given the test article daily beginning 42 days before cohabitation (maximum 14 days) and continuing through Day 20 of pre sumed gestation (rats assigned to Caesarean-sectioning), Day 24 of presumed Research Pathology Services, Inc. -1 001071 418-018:PAGE J-4 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS PROTOCOL 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 HISTOPATHOLOGY REPORT gestation (rats assigned to natural delivery that do not deliver a litter) or Day 4 post partum (rats that deliver a litter). Rats in the process of delivering were not given the test/control article. Male rats were used only as breeders and were not consid ered part of the test system. F1 generation pups were not directly given the test article, but were possibly exposed to the test article during maternal gestation (in utero exposure) or via maternal milk during the lactation period. Dosages of the FO generation rats were adjusted for the most recently recorded body weight and given at approximately the same times each day. On Day 5 postpartum, pups were sacrificed and the heart and thyroid were collected and preserved in 10% neutral buffered formalin. The in-life portion of the study, necropsies, and recording of the gross observations at necropsy were per formed by the staff of Argus Research Laboratories, Inc. Representative sections of the heart and thyroid of one male and one female F1 generation pup from the vehicle control group (Group 1) and 2 mg/kg PFOS group (Group 13) were routinely processed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin for microscopic examination. Upon completion of the. project, all raw data (remaining wet tissue, paraffin blocks, microscopic slides and histology records) will be returned to Argus Research Laboratories, Inc. for archiving. Research Pathology Services, Inc. -2 001072 418-018:PAGE J-5 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS PROTOCOL 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 HISTOPATHOLOGY REPORT RESULTS: The type and incidence of the histomorphologic observations in the heart and thyroid of the male and female pups from dams of the vehicle control and 2 mg/kg PFOS dosage groups are shown below. Dose Group: Sex: Dam Number: Number of Puds Examined: 1 M 10909 1 THYROID: No. Examined No. Normal 1 1 HEART: No. Examined No. Normal 1 1 13 M 11059 1 1 1 1 1 1 F 10909 1 13 F 11059 1 11 11 11 11 No microscopic changes were observed in the heart or thyroid of the male and female pups exposed to 2 mg/kg of PFOS. The sections examined were all within normal histologic limits. Research Pathology Services, Inc. -3 001073 418-018:PAGE J-6 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATIO N IN RATS PROTOCOL 418-018 S P O N S O R 'S STUDY NUMBER: T-6295.25 HISTOPATHOLOGY REPORT SUMMARY: Microscopic examination was made of the heart and thyroid from one male and one female F1 generation pup from dams of each of the vehicle control group (Group 1) and 2 mg/kg PFO S group (Group 13). No microscopic changes were seen in the heart or thyroid of the male and fe male pups exposed to 2 mg/kg of PFOS. R esearch Pathology S ervices, Inc. -4 001074 418-018:PAGE J-7 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE A ND NOEL INVESTIGATIO N IN RATS PROTOCOL 418-018 S P O N S O R 'S STU D Y NUMBER: T-6295.25 HISTOPATHOLOGY REPORT QUALITY ASSURANCE UNIT STATEMENT All aspects of the tissue processing, microscopic slide preparation, histopathologic evaluation and report preparation for the study listed above have been performed according to the Standard Operating Pro cedures of Research Pathology Services, Inc. and were audited in accordance with the procedures established by the Quality Assurance Unit of Research Pathology Services, Inc. in compliance with the Good Laboratory Practice regulations specified in the protocol. Study Design as Modification of: US Food and Drug Administration )1994). Intemation Conference on Harmonisation; Guide line on detection of toxicity to reproduction for medicinal products. Federal Register, September 22,1994, Vol. 59, No. 183. US Food and Drug Administration. Good Laboratory Practice Regulations: Final Rule. 21 CFR Part 58. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laoratory practive. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Or dinance Number 21, March 26,1997. Quality Assurance Unit study-based inspections were performed as shown below. There were no devia tions from the protocol, Standard Operating Procedures and/or appropriate Good Laboratory Practice regu lations noted during the conduct of the study. The summary report of QA inspections is included in the final re port submitted to the Study Director on December 13, 2002. Dates of Inspection Study Phase 04/09/01 04/09/01 04/24/01 05/01/01 06/07/01 06/08/01 07/13/01 07/17/01 07/25/01 07/27/01 07/30/01 08/03/01 08/08/01, 10/17/01 12/13/02 Master Schedule Pre-initiation Trimming Embedding Microtomy Staining Histopathology Data Entry Data Verification Data Processing Report Preparation Pre-submission Audit Draft report Final report Dates Reported to Date Reported Manaaement to Study Director 04/30/01 04/30/01 04/30/01 05/31/01 06/30/01 06/30/01 07/31/01 07/31/01 07/31/01 07/31/01 07/31/01 08/08/01 08/08/01, 10/17/01 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 12/13/02 4 Karen W. Harkins, BS Quality Assurance Unit 3- 15 Date R esearch Pathology Services, Inc. -5 001075 APPENDIX K HISTORICAL CONTROL DATA <m 076 SUMMARY OF REPRODUCTIVE INDICES C rl:C D (SD )IG S B R RATS DAY 21 C-SECTION 418-018:PAGE K-l PERIOD JUNE 1998 - SEPTEMBER 1999 NUMBER OF STUDIES 13 NUMBER OF RATS: TESTED PREGNANT FOUND DEAD ABORTED DELIVERED 252 235 1 0 0 NUMBER OF RATS PREGNANT AT CAESAREAN-SECTIONING 234 NUMBER OF RATS WITH SINGLE CONCEPTUS LITTER: LIVE RESORBED ABORTED 0 0 0 RANGE/STUDY MEAN or % MEAN or % % PREGNANT 93.4 (87.5-100) AVERAGE # CORPORA LUTEA 17.6 (16.4-19.1) AVERAGE # IMPLANTATIONS 14.9 (14.0-16.3) AVERAGE LITTER SIZE AVERAGE # LIVE FETUSES AVERAGE # DEAD FETUSES 14.5 (13.6-15.9) 0.0 - AVERAGE # RESORPTIONS 0.5 (0.1-1.4) AVERAGE # EARLY RESORPTIONS AVERAGE # LATE RESORPTIONS 0.5 0.0 (0.1-1.4) (0-0.1) 001077 418-018:PAGE K-2 SUMMARY OF REPRODUCTIVE INDICES C rl:C D (SD )IG S B R RATS DAY 21 C-SECTION MEAN or % RANGE/STUDY MEAN or % AVERAGE % DAMS WITH ANY RESORPTIONS 39.3 (12.5-71.4) AVERAGE % DAMS WITH ALL CONCEPTUSES RESORBED 0.0 - AVERAGE % DAMS WITH ONE OR MORE LIVE FETUSES 100.0 ~ AVERAGE SEX RATIO, (% MALES/LITTER) 49.4 (43.0-57.0) AVERAGE FETAL BODY W EIGHT (G) 5.28 (5.10-5.48) AVERAGE FOR MALES (G) 5.40 (5.15-5.63) AVERAGE FOR FEMALES (G) 5.12 (4.92-5.29) AVERAGE % DEAD OR RESORBED CONCEPTUSES/LITTER 3.3 (0.8-8.8) 00107# SUMMARY OF MATERNAL NECROPSY OBSERVATIONS C ri:C D (SD )IG S BR RATS -DAY 21 C-SECTION 418-018:PAGE K-3 PERIOD JUNE 1998 - SEPTEMBER 1999 # STUDIES # RATS TESTED # RATS PREGNANT # RATS DIED # RATS ABORTED # RATS DELIVERED # RATS WITH 100% RESORPTION 13 252 235 1 0 0 0 OBSERVATIONS LIVER Left lateral lobe, tan area UTERUS Moderate hydrometra INGUINAL AREA Subcutaneous, tan mass on right side R A N G E /S T U D Y N% N % 1 0.40 0-1 (0-4.0) 1 0.40 0-1 (0-4.2) 1 0.40 0-1 (0-4.5) 001079 SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS Crl:C D (SD )IG S BR RATS -DAY 21 C-SECTION 418-018:PAGE K-4 PERIOD JUNE 1998 - SEPTEMBER 1999 # STUDIES INCLUDED # LITTERS EXAMINED # LIVE FETUSES EXAMINED 13 234 3383 ALTERATION EYE(S) Eye bulges depressed JAWS Micrognathia HINDPAW Extra digit present TAIL Threadlike N% RANGE /STUDY N% L 1 0.43 F 1 0.03 0-1 (0-4.2) 0-1 (0-0.3) L 1 0.43 F 1 0.03 0-1 (0-4.2) 0-1 (0-0.3) L 1 0.43 F 1 0.03 0-1 (0-4.5) 0-1 (0-0.3) L 1 0.43 F 1 0.03 0-1 (0-4.3) 0-1 (0-0.3) L: LITTER INCIDENCE F: FETAL INCIDENCE 001080 418-018:PAGE K-5 SUMMARY OF FETAL SOFT TISSUE ALTERATIONS C h :C D (S D )IG S B R RATS DAY 21 C-SECTION PERIOD JUNE 1998 - SEPTEMBER 1999 # STUDIES INCLUDED 8 # LITTERS EXAMINED 180 # FETUSES EXAMINED 1245 BRAIN ALTERATION Dilation of lateral ventricles L F HEART Septal defect L F VESSELS Umbilical artery, descends to left of urinary bladder Innominate artery, absent Aorta passes dorsal to the trachea and esophagus Left carotid rises to the right of the right carotid Right carotid rises to the right of the right sub clavian Pulmonary artery, small Semilunar valve, small Ductus arteriosis attached to the left subclavian Great vessels, transposed L F L F L F L F L F L F L F L F L F LUNGS Apical and cardiac lobes, absent L F KIDNEY(S) Pelvis, slight dilation Pelvis, marked dilation L F L F RANGE/STUDY N% N% 2 1.11 0-2 (0-8.3) 2 0.16 0-2 (0-1.2) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 3 1.67 0-2 (0-9.1) 3 0.24 0-2 (0-1.2) 2 1.11 0-2 (0-8.3) 2 0.16 0-2 (0-1.2) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 2 1.11 0-1 (0-4.3) 2 0.16 0-1 (0-0.7) 1 0.56 0-1 (0-4.2) 1 0.08 0-1 (0-0.6) 1 0.56 0-1 (0-4.5) 1 0.08 0-1 (0-0.7) 1 0.56 0-1 (0-4.5) 1 0.08 0-1 (0-0.7) L: LITTER INCIDENCE F: FETAL INCIDENCE 001081 418-018:PAGE K-6 SUMMARY OF FETAL SKELETAL ALTERATIONS Crl:C D (SD )IG S B R RATS -DAY 21 C-SECTION PERIOD JUNE 1998 - SEPTEMBER 1999 # STUDIES INCLUDED 8 # LITTERS EXAMINED 180 # FETUSES EXAMINED 1332 ALTERATION SKULL Eye socket: small Mandibles: short VERTEBRAE Thoracic: Centrum, bifid : Centrum, fused to arch : 4 present Lumbar: Fused : 8 present : Centrum, fused to arches RIBS Cervical Rib(s) present One or more, wavy One or more, incompletely ossified (hypoplastic), or not ossified Fused 4 present R A N G E /S T U D Y N% N% L 1 0.56 F 1 0.08 L 1 0.56 F 1 0.08 0-1 (0-4.2) 0-1 (0-0.6) 0-1 (0-4.2) 0-1 (0-0.6) L 4 2.22 F 4 0.30 L 1 0.56 F 1 0.08 L 1 0.56 F 1 0.08 L 2 1.11 F 2 0.15 L 1 0.56 F 1 0.08 L 1 0.56 F 1 0.08 0-1 (0-4.8) 0-1 (0-0.6) 0-1 (0-4.8) 0-1 (0-0.6) 0-1 (0-4.8) 0-1 (0-0.6) 0-1 (0-4.8) 0-1 (0-0.6) 0-1 (0-4.8) 0-1 (0-0.6) 0-1 (0-4.8) 0-1 (0-0.6) L 12 6.67 F 14 1.05 L 6 3.33 F 8 0.60 L 2 1.11 F 3 0.22 L 1 0.56 F 1 0.08 L 1 0.56 F 1 0.08 0-5 (0-20.8) 0-6 (0-3.5) 0-2 (0-9.5) 0-3 (0-1.8) 0-2 (0-9.1) 0-3 (0-1.8) 0-1 (0-4.2) 0-1 (0-0.6) 0-1 (0-4.8) 0-1 (0-0.6) L: LITTER INCIDENCE F: FETAL INCIDENCE 001082 418-018:PAGE K-7 SUMMARY OF FETAL SKELETAL ALTERATIONS Crt:CD(SD)IG S B R RATS -DAY 21 C-SECTION ALTERATION MANUBRIUM Duplicated STERNEBRAE One or more incompletely ossified or not ossified Asymmetric Duplicated XIPHOID Duplicated HINDLIMB Extra digit present Femur, bent L: LITTER INCIDENCE F: FETAL INCIDENCE R A N G E /S T U D Y N% N% L 1 0.56 F 1 0.08 0-1 (0-4.5) 0-1 (0-0.6) L 4 2.22 F 4 0.30 L 2 1.11 F 2 0.15 L 1 0.56 F 1 0.08 0-2 (0-9.1) 0-2 (0-1.2) 0-1 (0-4.5) 0-1 (0-0.6) 0-1 (0-4.5) 0-1 (0-0.6) L 1 0.56 F 1 0.08 0-1 (0-4.5) 0-1 (0-0.6) L 1 0.56 F 1 0.08 L 1 0.56 F 1 0.08 0-1 (0-4.5) 0-1 (0-0.6) 0-1 (0-4.8) 0-1 (0-0.6) 001083 SUMMARY OF FETAL OSSIFICATION SITES SKELETAL AVERAGES Crl:CD(SD)IGSBR RATS DAY 21 C-SECTION 418-018.-PAGE K-8 PERIOD: JUNE 1998 - SEPTEMBER 1999 # STUDIES INCLUDED 8 # LITTERS EXAMINED 180 # FETUSES EXAMINED 1331 SKELETAL AVERAGES HYOID VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS (pairs) STERNUM MANUBRIUM STERNAL CENTERS XIPHOID FOREPAWS (Calculated as average per limb) CARPALS METACARPALS DIGITS PHALANGES HINDPAWS (Calculated as average per limb) TARSALS METATARSALS DIGITS PHALANGES FETUS/LITTER MEAN RANGE/STUDY 0.99 (0.96-1.00) 7.00 13.07 5.92 3.00 7.44 13.05 1.00 3.99 1.00 (13.03-13.14) (5.85-5.97) (7.11-7.63) (13.02-13.12) ~ (3.98-4.00) -- 0.00 4.00 5.00 8.28 (3.99-4.00) (8.04-8.52) 0.02 4.80 5.00 6.21 (0-0.07) (4.73-4.87) - (5.81-6.54) 001084 APPENDIX L STATEMENT OF THE STUDY DIRECTOR 001085 418-018-.PAGE L -l Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215)443-8587 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACED/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 STATEMENT OF THE STUDY DIRECTOR. This final report accurately reflects the raw data obtained during the performance of the study. No deviations from the U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations; Final Rule3, the Japanese Ministry of Health and Welfare (MHW) Good Laboratory Practice Standardfor Safety Studies on Drugsb and the Organisation for Economic Co-operation and Development (OECD). The Revised OECD Principles o f Good Laboratory Practices'^ occurred that affected the quality or integrity of the study, with the exceptions in the bulleted list bellow: There were two study directors for this study; one for the in-life phase and one for the analytical phase. The in-life study phase was considered to end at the generation and shipment of specimens. The analytical study phase was considered to start at the receipt of these specimens for analysis. Since the technical performance of each phase was entirely separate, no effect is expected from this exception. Automated data collection systems have not been validated as required according to 21 CFR 58.130(c). The hard copy printout is considered the raw data. The stability of the reference standard and internal standard were not determined. if * ' IkDfJDZ- Raymond G. York, Pfr.D., )ABT Associate Director of Restiarch and Study Director Argus Research Laboratories, Inc. Date a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standardfo r Safety Studies on Drugs. Ordinance No. 21, March 26, 1997, Japan. c. Organisation for Economic Cooperation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final], 01<J8<> APPENDIX M QUALITY ASSURANCE STATEMENT 001087 418-018:PAGE M-l Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215)443-8587 QUALITY ASSURANCE STATEMENT Argus Protocol: 418-018 Study Director: Raymond G. York, Ph.D., DABT Sponsor's Study Number: T-6295.25 The protocol, critical phases, raw data and final report were inspected by the Quality Assurance Unit (QAU), to assure conformance with: U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standardfor Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles o f good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. The undersigned indicate that the report is an accurate representation of the raw data. Data provided by the Sponsor or a subcontractor were not audited by the Primedica Argus Quality Assurance Unit. The draft protocol for this study was audited for adherence to the appropriate regulations(s), as stated above, on 27 JUN 00, 31 JUL 00, 15 AUG 00 and 21 AUG 00. The QAU inspection and report audit dates are listed below: Inspection Phase Inspection Date(s) Date(s) Findings Date(s) Findings Submitted to Study Submitted to Director Management Test Article Preparation Blood Collection Caesarean-Sectioning Tissue Collection Natural Delivery/Litter Dam/Litter Sacrifice Milk Collection 04 OCT 00 31 OCT 00 31 OCT 00 31 OCT 00 02 NOV 00 06 NOV 00 06 NOV 00 11 OCT 00 31 OCT 00 03 NOV 00 03 NOV 00 03 NOV 00 08 NOV 00 08 NOV 00 11 OCT 00 31 OCT 00 03 NOV 00 03 NOV 00 03 NOV 00 08 NOV 00 08 NOV 00 001088 418-018:PAGE M-2 Observations Inspection Phase In-Life Raw Data Formulations Raw Data Report Tables Report Text Revised Report Inspection Date(si 18 JAN 01, 19 JAN 01, 22 JAN 01 to 26 JAN 01 and 29 JAN 01 to 31 JAN 01 01 FEB 01, 05 FEB 01, 07 FEB 01 to 09 FEB 01 and 12 FEB 01 to 19 FEB 01 25 FEB 01 to 28 FEB 01 25 FEB 01 to 26 FEB 01 and 23 MAR 01 to 29 MAR 01 17 MAR 01, 30 MAR 01 and 02APR 01 to 03 APR 01 to 10 APR 01, 12 APR 01, 13 APR 01 and 16 APR 01 and 17 APR 01 18 APR 01 04 MAY 01 and 07 MAY 01 17 JUL01 10 OCT 01 30 OCT 01 31 JAN 02 04 FEB 02 27 MAR 02 10 DEC 02 Date(s) Findings Date(s) Findings Submitted to Study Submitted to Director__________ Management 01 FEB 01 01 FEB 01 19 FEB 01 19 FEB 01 01 MAR 01 29 MAR 01 03 APR 01 16 APR 01 17 APR 01 18 APR 01 07 MAY 01 17 JUL01 10 OCT 01 31 OCT 01 31 JAN 02 04 FEB 02 27 MAR 02 10 DEC 02 01 MAR 01 29 MAR 01 03 APR 01 16 APR 01 17 APR 01 18 APR 01 07 MAY 01 17 JUL01 10 OCT 01 31 OCT 01 31 JAN 02 04 FEB 02 27 MAR 02 10 DEC 02 Mafthew J. Vaneman, B.S. Date Manager, Regulatory Compliance \ n s m i t n 6 i h n n it> t c Cynthia M. Kelsch Quality ssurance Associate and Principal Auditor 001089 Date