Document ZnV6Mgxrer66v6brG38Y10zJ0

frR -2 6 _ $ 1 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 000433 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 1-10 n. 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 -11 m. RESULTS m -i A. Mortality m -i B. Clinical Observations C. Necropsy Observations 0434 ii m -i m -2 SUBJECT PAGE D. Terminal Body Weights and Organ Weights and Ratios of Organ Weights to Terminal Body Weights ID-2 E. Body Weights and Body Weight Changes HI-3 F. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values III-5 G. Mating and Fertility EU-7 H. Caesarean-Sectioning and Litter Observations HI-7 I. Natural Delivery and Litter Observations HI-7 J. Clinical Observations from Birth to Day 5 Postpartum and Necropsy Observations ni-8 K. Blood and Tissue Analyses HI-9 L. Histopathology of Hearts and Thyroids F I Generation Pups HI-27 REFERENCES DI-28 APPENDIX A - REPORT FIGURES Figure 1. Body Weights - Fo Generation Female Rats - Groups 1,8 through 13 A -l Figure 2. Body Weights - Fo Generation Female Rats - Groups 2,3 and 4 A-2 Figure 3. Body Weights - Fo Generation Female Rats - Groups 5,6 and 7 A-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 iii 000435 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 - FoGeneration 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 00043G 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 000437 V 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 - F I 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) 1-1 to 1-444 APPENDIX J - HISTOPATHOLOGY REPORT J-l to J-7 APPENDIX K - HISTORICAL CONTROL DATA K -lto K-8 APPENDIX L - STATEMENT OF THE STUDY DIRECTOR L-l APPENDIX M - QUALITY ASSURANCE STATEMENT M -l to M-2 000438 vi 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 no observed-effect-level (NOEL). A. Methods'* 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). 000439 The dosage groups and dosing schedule were as follows: 418-018.PAGEI-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 H20 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 H20 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid | | Not applicable | Not applicable 1 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 0.5% Tween 80 Mevalonic Acid Cholesterol PFOS PFOS PFOS PFOS 0 0 1000 500 0.4 0.8 1 1.2 0 0 100 100 0.08 0.16 0.20 0.24 PFOS PFOS 1.6 2 0.32 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. 0004.40 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 o f 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 000441 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 o f 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. o O \ *j ,< 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. 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 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 000443 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 000444 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 000445 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 1 2 ,1 3 ,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 o f 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. 000446 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 T3 levels 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 T4 were 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 U0044'?' 418-018:PAGE 1-10 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 T3and 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). / Associate Director of Research & 6 ^ 72Date / (e Associate Director of Research and Study Director loQZ- Date 000448 418-018:PAGE II-1 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.7. Regulatory Compliance The requirements of the U.S. Food and Drug Administration (FDA)(,) 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)(2), the Japanese Ministry of Health and Welfare (MHW)(3) and the European Economic Community (EEC)W. 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 U -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 000450 418-018:PAGE IL-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 DLb 4 (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 0 0 -1 8 NOV 00 09 OCT 00 PM -16 OCT 00 AM 16 OCT 00 PM - 23 OCT 00 AM 31 OCT 0 0 -0 3 NOV 00 05 NOV 00 - 09 NOV 00 17 NOV 00 31 OCT 0 0 - 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. OOOhS I 418-018:PAGE n-4 A . 17. Records M aintained 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 N um ber 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 000452 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 (R O D IH 2O). 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. V 000453 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 | Homogeneity" Concentration 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 Frozen Frozen Room temperature Testing Facility Archives 01 SEP 00 01 SEP 00 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 o f study: premating, gestation and postnatal. Each sample was divided into two aliquots, one o f 2 mL and one o f 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. 000^54 418-018:PAGE II-7 E.2. A nalytical 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/PIus 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/PIus 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. 0004S5 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 Arrival3 Weight (g) at Study Assignment Replicate 1 180 27 JU N 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 Shipment 2 150 150 29 FEB 00 20 MAR 00 71 days 72 days 245 - 339 286 - 344 473 - 767 537 - 748 452 - 773 593 - 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 ,1 2 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. 000456 418-018:PAGE n-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 e ts e q . 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 G u id e f o r the C a re a n d Use o f L aboratory A n im als^ . 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). 000457 418-018:PAGE H-10 G.6. Feed Rats were given a d lib itu m 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 a d libitu m 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. 000453 H. Methods H .l. Dosage Administration The dosage groups and dosing schedule were as follows2. 418-018:PAGE n i 1 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 R O D I H 20 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 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 9 0.8 mg/kg PFOS Not Applicable 0.8 mg/kg PFOS 10 1 mg/kg PFOS Not Applicable 1 mg/kg PFOS 11 1.2 mg/kg PFOS Not Applicable 12 1.6 mg/kg PFOS Not Applicable 13 2 mg/kg PFOS Not Applicable 1.2 mg/kg PFOS 1.6 mg/kg PFOS 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. 000459 418-018:PAGE 11-12 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 G roup Identification 1 Vehicle Control M evalonic Acid 2 C o n tro l 1.6 m g/kg PFOS + 3 M evalonic Acid 4 2 mg/kg PFOS + M evalonic Acid 5 Cholesterol Control 1.6 m g/kg PFO S + 6 C h o lestero l 2 mg/kg PFOS + 7 Cholesterol 8 0.4 mg/kg PFOS 9 0.8 mg/kg PFOS 10 1 m g/kg PFO S 11 1.2 m g/kg PFO S 12 1.6 m g/kg PFO S 13 2 m g/kg PFO S Number of Rats 28 28 29 28 28 28 28 20 20 20 20 28 28 Assigned Rat Numbers Replicate 1 Replicate 1 C aesarean -sectio n in g Natural Delivery 10901 - 10905,10907, 10908,10910,10912 10915 - 10920, 10925, 10928 10930-10935, 10937, 10940 1 0 9 4 5 -1 0 9 4 7 ,1 0 9 4 9 10951,10953,10954 10958- 10965 10972, 10974- 10976, 10978, 10980- 10982 10906,10909, 10911, 10913,10914 10921 - 10924, 10926, 10927 10929,10936,10938, 10939, 10941, 10942 10943, 10944, 10948, 10952,10955,10956 10957, 10966- 10970 10971, 10973, 10977, 10979,10983, 10984 10985, 10987- 10993 Not applicable Not applicable Not applicable Not applicable 11040, 11044- 11047, 11050- 11052 11053, 11054, 11056, 11058, 11060, 11062, 11063, 11066 10986, 10994- 10998 10999- 11008 11009- 11018 11019-11028 11029- 11038 11039, 11041 - 11043, 11048, 11049 11055, 11057, 11059, 11061, 11064, 11065 Replicate 2 Natural Delivery 11501 -11 5 1 4 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. 000460 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 u tero 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 APPENDIX D, items 8 and 9. U004C1 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)3. 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 sacrifice3. Feed consumption values were recorded weekly to cohabitation, on DGs 0 ,7 ,1 4 ,2 1 and 25 (if necessary) and DLs 1 and 5 (rats assigned to natural delivery)b. 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. F I 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. 00046 418-018:PAGE 11-15 H.6. Gross Necropsy* H.6.a. Fo Generation Rats Assigned to Caesarean-Sectioning Eight rats from Groups 1 to 7 ,1 2 and 13 (Replicate 1) were sacrificed by carbon dioxide asphyxiation on DG 21. Blood and liver samples were collected from these rats6. 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 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. 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 recordedc. 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*5. 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 APPENDDC D, item 12. c. See APPENDIX D, item 13 d. See APPENDIX D, item 14. 000463 418-018:PAGE 11-16 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 APPENDIX D, item 12. ^ 000464 418-018:PAGE 11-17 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. 00Q465 418-018:PAGE 11-18 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 o f 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 APPENDIX D, item 16. 000466 418-018:PAGE 11-19 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, triglycerides* Plasma <-20C Primedica Worcester Mevalonic acid (Groups 1,2, 3 ,4 ,1 1 , 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, triglycerides* Pooled Plasma <-20C Primedica Worcester Mevalonic acid (Groups 1 ,2 ,3 ,4 ,1 1 , 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, Plasma <-20C Primedica Worcester triglycerides* 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. T he first priority w as analysis for total cholesterol and glucose. T he second priority w as for total and fre e T 3, T 4 a n d T S H le v e ls . T h e th ird p r io r ity w a s a n a ly s is f o r L D L , H D L a n d tr ig ly c e rid e s . 000467 418-018:PAGE 0-20 Sample Storage/Shipping Conditions Recipient Analysis Pups Replicate 1 Serum - aliquot 1 Serum - aliquot 2 <-70C <-70C Sponsor AniLytics PFOS* Total cholesterol, glucose, total & free T3, T4,TSH (Groups 1, 11, 12,13), LDL, HDL, triglycerides6 Plasma <-70C Primedica Worcester Mevalonic acid (Groups 1,2, 3,4 , 11,12 and 13) Pups - Replicate 2 Serum - aliquot 1 <-70C Sponsor PFOS* 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 Liver - 1/3 pooled livers Gluteraldehyde <-70C Sponsor Sponsor Possible EM 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 females per litter0 Gluteraldehyde Sponsor Possible EM and light microscopy Pooled Thyroids*1 10% neutral buffered formalin Testing Facility Possible histopathology 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 o f one male and one female FI generation pup from each o f 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 o f 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 P rim e d ic a A rg u s A u to m a te d D a ta C o lle c tio n a n d M a n a g e m e n t S ystem and the V ivarium T e m p e ra tu re a n d R e la tiv e H u m id ity M o n ito r in g S y ste m . All data were tabulated, summarized and/or statistically analyzed using the P rim ed ica A rg u s A u to m a ted D a ta C o llectio n a n d M a n a g em en t S ystem , the V ivariu m T em peratu re a n d R ela tive H u m idity M o n ito r in g S ystem , M ic r o s o ft E x c e l [part o f Microsoft Office 97 (version SR-2)] and/or The SA S S ystem (version 6.12). 000468 418-018:PAGE 11-21 The following schematic represents the statistical analyses of the data: Type of Test8 L Parametric A. Bartlett's Testc I Significant at p<0.001 I Not Significant II. Nonparametricb A. Kruskal-Wallis Test (<75% ties) Significant at /j^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 in. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilities are reported as either p < 0 .0 5 or p<0.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. 0004C9 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 and the Analysis of Variance(12), when appropriate [i.e., Bartlett's Test was not significant (p>0.001)]. If the Analysis of Variance was significant (p<0.05), Dunnett's Test(13) 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 Test(I4) 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 Test(16) 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 Test(14). u<m?o 418-018:PAGEm-l 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 .0 1 ) 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 0004?! 418-018:PAGE ID-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 (p < 0.05 or p<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 (p < 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 (p < 0 .0 5 or p<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 (p<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 (p<0.05 or p<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 p<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 000472 418-018:PAGEm-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 (><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 (><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 (><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 (><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 (><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 (7<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 (><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 (>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 000473 418-018:PAGE IH-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 p<0.01) in Groups 9 ,1 0 , 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<0.05 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. 000474 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 (p < 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 ><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 p < 0 .0 l ) 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 /?<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 (><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<0.0 1) 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 (p < 0.05 or ><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 (><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). 000475 418-018:PAGE m -6 F.2. Gestation Absolute and relative feed consumption values were significantly reduced (p< 0.05 or p < 0 .0 l ) 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 {p < 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 .0 5 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 o f 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 (p<0.01) in Groups 6 and 7 on DGs 0 to 7 and significantly increased (p < 0.05 or p<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 (p < 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 .0 l) 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). 000476 418-018:PAGE ffl-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 Facility3, 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). 000477 418-018:PAGE IH-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 p<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 (p<0.05 or p<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 p<0.01) in Groups 1 2 ,1 3 ,4 ,6 and 7. The numbers of surviving pups per litter were significantly reduced (p< 0.05 or p<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 (p < 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 ,1 0 ,1 1 ,1 2 and 13 (0.4,0.8, 1, I. 2,1.6 and 2 mg/kg/day PFOS alone, respectively) were also significantly reduced (p < 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 (p<0.01) in Group 8 (0.8 mg/kg PFOS alone) and significantly reduced (p<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 (p<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 p<0.01 in Group 7. 000478 418-018:PAGE ffl-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 ,1 0 ,1 1 , 12 and 13, reflecting the single occurrence o f 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/dL) of PFOS alone Param eter Cholesterol G lu co se L D L -D ir H D L -D ir Group 1 (V ehicle) G roup 12 84.3 13.3 (8 ) 78.1 10.5 98.9 + 11.7 (8 ) 104.9 7.8 7.1 5 .0 (8 ) 10.8 4 .4 43.3 1 9 .0 (8 ) 43.5 11.2 1 . 6 mg/kg G roup 13 (8 ) 72.3 6 .3 (8 ) 96.3 18.0 (8 ) 11.3 3 .6 (8 ) 41.0 11.7 2 mg/kg (8 ) (8 ) (8 ) (8 ) Triglycerides 313.5 233.6 (8 ) 194.3 65.2 (8 ) 201.0 87.5 (8 ) M evalonic 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. 000479 418-018:PAGE III-10 Thyroid Hormone Values of PFOS a one P aram eter Total T4 ug/dL Total T 3 ng/dL FreeT 3 p g/m L Group 1 0 .5 8 4 + 0 .4 8 2 73.979 21.234 0.579 0.209 (V ehicle) (8 ) (8 ) (8 ) G roup 12 1 . 6 mg/kg 0 . 0 0 0 0 .0 0 0 ** (8 ) 47.503 10.268** (8 ) 0.140 0.117** (7) G roup 13 2 mg/kg 0 . 0 0 0 0 .0 0 0 ** (8 ) 42.440 8.271** (8 ) 0.157 0.161** (7) TSH n g/m L 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 (p < 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 L D L -D ir H D L -D ir Group 1 (V ehicle) G roup 12 1 . 6 mg/kg G roup 13 ____ 2. m frg ____ 3.0 0.38 (8 ) 2 . 6 0 .2 2 * (8 ) 2.4 0.26** (8 ) 0.5 0 .2 7 (8 ) 0.7 0.24 (8 ) 0 . 6 0 .1 2 (8 ) 0.5 0.12 (8 ) 0 . 6 0 .1 0 (8 ) 0.5 0.06 (8 ) Triglycerides 8 . 0 1 .6 6 (8 ) 7.7 0.71 (8 ) 6 . 6 0.82 (8 ) The clinical chemistry parameter of low density lipoprotein (LDL) was significantly increased (p<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 Param eter Cholesterol G lu co se L D L -D ir H DL-D ir Triglycerides M evalonic Acid Group 2 (M evalonic) Group 3 1 . 6 mg/kg Group 4 2 mg/kg 92.5 14.6 (8 ) 91.0 21.4 (8 ) 88.9 23.1 (8 ) 108.5 11.9 (8 ) 108.1 15.4 (8 ) 106.0 14.0 (8 ) 2.9 3.6 (8 ) 11.8 9.0** (8 ) 9.6 5.0* (8 ) 32.4 20.0 (8 ) 35.1 12.1 (8 ) 47.8 16.2 (8 ) 453.9 307.8 (8 ) 305.9 + 85.1 (8 ) 243.6 270.8 (8 ) 1663.8 4 1 5 .6 (8 ) 1369.8 240.1 (8 ) 1482.5 5 2 1 .9 (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 (p<0.05 or /?<0.01), compared to the vehicle control group. Significantly different from control group; p < 0.05. Significantly different from control group; /?<0.01. 0004S0 418-018:PAGE ffl-11 Thyroid Hormone Values of PFOS plus Mevalonic Acid Parameter Total T4 ug/dL Total T3 ng/dL FreeTj pg/mL TSH ng/mL Group 2 (Mevalonic) 0.414 0.510 (8 ) 73.571 14.314 (8 ) 0.514 0.203 (7) 2.627 1.177 (6 ) Group 3 1 . 6 mg/kg Group 4 2 mg/kg 0.003 0.007* (8 ) . **0 . 0 0 0 0 000 (8 ) 54.043 6.670** (8 ) 46.408 8.895** (8 ) 0.313 0.146* (8 ) 0.184 0.170** (8 ) 1.889 0.391 (7) 2.375 1.086 (8 ) FreeT4 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 Group 4 (8 ) 2.4 0.30** (8 ) 0 . 8 0 . 1 2 ** (8 ) 0.6 0.05 (8 ) 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 Chemistry Values (mg/dL) of PFOS plus Cholesterol Parameter Cholesterol Glucose LDL-Dir HDL-Dir Group 5 (Cholesterol) Group 6 1 . 6 mg/kg Group 7 89.6 19.6 (8 ) 79.3 9.4 (8 ) 77.9 11.5 100.4 20.2 (8 ) 98.5 18.5 (8 ) 106.4 17.9 8.5 5.2 (8 ) 9.9 3.9 (8 ) 1 2 . 0 8 .6 28.0 11.5 (8 ) 41.0 13.6 (8 ) 46.6 18.4 2 mg/kg (8 ) (8 ) (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 .0 \. 000461 418-018:PAGE HI-12 Thyroid Hormone Values of PFOS plus Cholesterol Parameter Total T4 ug/dL Total T3 ng/dL FreeT3 pg/mL Group 5 0.540 0.529 76.900 21.899 0.694 0.256 (Cholesterol) (8 ) (8 ) (7) Group 6 1 . 6 mg/kg 0.013 0.035** 52.031 9.331** 0.235 0.123** (8 ) (8 ) (8 ) Group 7 2 mg/kg . **0 . 0 0 0 0 000 (8 ) 49.020 8.527** (8 ) 0.246 0.180** (8 ) TSH ng/mL 2.544 0.759 (5) 1.452 0.691 (6 ) 2.013 0.854 (6 ) FreeT4 ng/dL 0.281 0.125 (7) 0.134 0.086** (8 ) 0 . 1 1 0 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 Values (mg/gm) o f 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 .1 0 7.7 0.72** 1 . 6 mg/kg Group 7 (8 ) 2.7 0.21 (8 ) 0 . 8 0.18** (8 ) 0.6 0.09 (8 ) 7.5 0.70** 2m &^g (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 000432 418-018:PAGE III-13 PFOS Levels in DG 21 Dam Sera and Liver Sera* Liver* 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 mgflcg 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 o f cholesterol and low density lipoprotein (LDL) were significantly increased (p < 0.05 or /?<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 0004S3 418-018:PAGE III-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 Parameter Total T4 ug/dL Total T3 ng/dL Free T3 pg/mL Group 1 (Vehicle) 0 . 0 0 0 0 .0 0 0 (7) 0.423 0.845 (4) 0 . 0 0 0 0 .0 0 0 (0 ) Group 12 0 . 0 0 0 0 .0 0 0 0 . 0 0 0 0 .0 0 0 0 . 0 0 0 0 .0 0 0 1 . 6 mg/kg Group 13 2 mg/kg (8 ) 0 . 0 0 0 0 .0 0 0 (8 ) (8 ) 0.925 1.147 (6 ) (1 ) 0 . 0 0 0 0 .0 0 0 (0 ) TSH ng/mL 0 . 0 0 0 0 .0 0 0 (0 ) 0 . 0 0 0 0 .0 0 0 (0 ) 1.160 0 . 0 0 0 (1 ) FreeT4 ng/dL 0.050 0.000 (1 ) . **0 . 0 0 0 0 000 (3) . **0 . 0 0 0 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 (Vehicle) 3.4 0.38 (8 ) . 0 0 0 0 0 . 0 0 0 0 (8 ) 0.01 0.009 (8 ) Group 12 1 . 6 mg/kg 3.4 0.38 (8 ) . 0 0 0 0 0 . 0 0 0 0 (8 ) 0 . 0 2 0.006* (8 ) Group 13 3.2 0.34 0 . 0 0 0 0 .0 0 0 0 0.02 0.015* 2 mg/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 .0 \. 000464 418-018:PAGE III-15 Clinical C emistry Va ues (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 (8 ) 29.4 5.3 (8 ) 13.9 1.2 (8 ) 36.4 8.7 (8 ) Group 3 1 . 6 mg/kg 68.3 13.4** 69.6 6.2 48.3 8.1** 16.1 3.0 (8 ) (8 ) (8 ) (8 ) 31.8 7.1 (8 ) Group 4 2 mg/kg 63.1 11.7 (8 ) 74.5 12.3 46.9 9.4** 15.5 1.9 25.1 8.1** (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 (p<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 Parameter Total T4 ug/dL Total T3 ng/dL FreeT3 pg/mL TSH ng/mL Group 2 (Mevalonic) 0 . 0 0 0 0 .0 0 0 (7) 2.810 4.867 (3) (0 ) (0 ) Group 3 1 . 6 mg/kg Group 4 2 mg/kg 0 . 0 0 0 0 .0 0 0 (8 ) 0 . 0 0 0 0 .0 0 0 (8 ) 1.096 1.379 (7) 2.928 2.747 (6 ) 0 . 0 0 0 0 .0 0 0 (1 ) 0 . 0 0 0 0 .0 0 0 (2 ) (0 ) (0 ) Free T4 ng/dL 0.070 0.000 (1 ) . **0 . 0 0 0 0 000 (2 ) . **0 . 0 0 0 0 000 (4) Liver cholesterol, HDL and triglyceride levels were significantly increased (p < 0 .0 5 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 Values (mg/gm) of PFOS plus Mevalonic Acid Parameter Cholesterol LDL-Dir HDL-Dir Triglycerides Group 2 3.6 0.49 . 0 0 0 0 0 . 0 0 0 0 0.01 0.009 5.4 1.25 (Mevalonic) (8 ) (8 ) (8 ) (8 ) Group 3 1 . 6 mg/kg 4.5 0.48** (8 ) . 0 0 0 0 0 . 0 0 0 0 (8 ) 0.04 0.019** (8 ) 8 . 0 1.16** (8 ) Group 4 5.1 0.82** 0 . 0 0 0 0 .0 0 0 0 0.03 0.018* 9.6 2.39** 2 mg/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. UQ04S5 418-018:PAGE III-16 Clinical C emistry Values (mg/dL' o f 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 T4was 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 plus Cholesterol Parameter Total T4 ug/dL Total T3 ng/dL Free T3 pg/mL TSH ng/mL Group 5 (Cholesterol) Group 6 1 . 6 mg/kg Group 7 2 mg/kg 0 . 0 0 0 0 .0 0 0 (8 ) 0 . 0 0 0 0 .0 0 0 (7) 0 . 0 0 0 0 .0 0 0 (6 ) 1.143 1.306 (6 ) 0.760 1.389 (5) 0.653 1.132 (3) 0 . 0 0 0 0 .0 0 0 (2 ) 0 . 0 0 0 0 .0 0 0 (2 ) 0 . 0 0 0 0 .0 0 0 (1 ) 0 . 0 0 0 0 .0 0 0 (0 ) 0.040 0.000 (1 ) 0 . 0 0 0 0 .0 0 0 (0 ) Free T4 ng/dL 0.037 0.015 (3) . **0 . 0 0 0 0 000 (4) . **0 . 0 0 0 0 000 (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 (Cholesterol) Group 6 1 . 6 mg/kg Group 7 2 mg/kg 3.6 0.29 (8 ) 4.4 0.40** (8 ) 4.0 0.68 (8 ) . 0 0 0 0 0 . 0 0 0 0 (8 ) . +0 0 0 0 0 . 0 0 0 0 (8 ) 0 . 0 0 0 0 .0 0 0 0 (8 ) 0.02+0.019 (8 ) 0.03 0.010 (8 ) 0.04 0.023 (8 ) 5.5 0.73 (8 ) 7.8 1.23** (8 ) 7.1 1.15** (8 ) PFOS concentrations were significantly increased (p<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<0.01. 0Q048G 418-018:PAGE HI-17 PFOS Levels in Pooled DG 21 Fetal Sera and Livers Sera* Liver* Parameter (ug/mL) (ppm) Group 1 (Vehicle) Group 2 (Mevalonic) 0.0 0.00 (6) 0.1 0.08 (6) N.A. 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 .0 3 (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 2 mg/kg 169.8 37.91** (6) N.A. N.A. = Not Analyzed 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 (/?<0.05 or /?<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; p<0.01. 0004S7 418-018:PAGE HI-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 T4levels in lactating dams administered 0.4 (free T4 only), 0.8, 1, 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced (p<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 Parameter Total T4 ug/dL Total T3 ng/dL FreeT3 pg/mL Group 1 1.454 0.660 74.674 18.977 0.638 0.297 (Vehicle) (19) (19) (19) Group 8 0.4 mg/kg 0.808 0.414 GO) 72.912 13.470 GO) 0.698 0.246 (1 0 ) Group 9 0.596 0.443* 63.780 6.684 0.588 0.152 0 . 8 mg/kg Group 10 (8 ) 0.728 0.243** (8 ) 62.348 13.180 (8 ) 0.552 0.230 1 mg/kg (9) (9) (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 mg/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) FreeTi ng/dL 0.788 0.219 (19) 0.462 0.114** GO) 0.279 0.060** (8 ) 0.277 0.095** (9) 0.260 0 .1 2 2 ** (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. 000488 418-018:PAGEHI-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 0 . 8 mg/kg 2.4 0.59 (17) 0.05 0.093 (17) 0.23 0.130 (17) Group 10 2.5 0.54 0.00 0.004 0.20 0.093 1 mg/kg Group 11 (19) 2.5 0.83 (19) 0.05 0.106 (19) 0.26 0.135 1 . 2 mg/kg (18) (18) (18) Group 12 1 . 6 mg/kg Group 13 2.6 0.79 (17) 2.9 1.08 0.01 0.038 (17) 0.03 0.054 0.21 0.104 (17) 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) 1 1 . 8 1 2 .0 2 (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 Values (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 Acic Total T4 Total T3 FreeT3 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. 000469 418-018:PAGE HI-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 (p<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 [Blood] [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 20.8 4.4 2.8 48.6 11.1 1 . 6 mg/kg (2 0 ) (1 2 ) (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 /?<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 Parameter Total T4 ug/dL Total T3 ng/dL Free T3 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* 2 mg/kg (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. 000490 418-018:PAGE 111-21 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) Group 6 2.3 0.69 0.01 0.036 0.20 0.115 1 . 6 mg/kg (2 0 ) (2 0 ) (2 0 ) Group 7 2.3 0.61 0 . 0 0 0 .0 0 0 0.19 0.078 2 mg/kg (19) (19) (19) Triglycerides 7.6 1.68 (17) 10.1 4.68 (2 0 ) 11.0 4.83* (19) PFOS Levels in DL 5a Dam Sera and Livers Sera" Liver6 Parameter Group 1 (ug/mL) 0.0 + 0.01 (ppm) 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 Group 11 12 Group 12 1.6 mg/kg Group 13 2 mg/kg 52.3 26.02* (6) 86.0 10.22* (6) 169.0 32.12* (6) 134.0 26.60* (6) N.A. N.A. 110.2 13.43* (5) 145.8 20.18* (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. 000491 418-018:PAGE ffl-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) (1 2 ) (1 2 ) (1 2 ) Group 8 111.9 + 11.3 244.5 60.1 29.1 7.4 0.4 mg/kg (15) (15) (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 m/k (3) (3) (3) HDL-Dir 29.8 7.9 (1 2 ) 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 (p<0.01), compared to the vehicle control group. The thyroid stimulating hormone (TSH) was significantly increased (p < 0.05 or /?<0.01) at 1 and 1.6 mg/kg PFOS, but not in a dosage-dependent fashion. Thyroid Hormone Values of PFOS alone Parameter Total T ug/dL Total T3 ng/dL Group 1 0.543 0.221 54.383 17.965 (Vehicle) (1 2 ) (1 2 ) Group 8 . **0 . 0 0 0 0 000 55.777 18.934 0.4 mg/kg GO) (9) Group 9 . **0 . 0 0 0 0 000 48.825 17.720 0 . 8 mg/kg (8 ) (8 ) Group 10 1 mg/kg 0.017 0.050** (9) 47.710 8.613 (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 2 mg/kg . **0 . 0 0 0 0 000 (3) 32.777 12.162 (3) Free T3 pg/mL 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 1 0 0.006** (6 ) . **0 . 0 1 0 0 010 (3) Liver triglyceride and LDL levels were decreased or significantly decreased (p<0.05 or p<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. UOCH92 418-018:PAGE IEL-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 0.4 mg/kg 3.9 0.48 (2 0 ) . **0 . 0 0 0 0 0022 (2 0 ) 0.09 0.031 (2 0 ) Group 9 3.8 0.30 . **0 . 0 0 0 0 0000 0.09 0.038 0 . 8 mg/kg (17) (17) (17) Group 10 4.0 0.83 . **0 . 0 0 0 0 0000 0.08 0.035 1 mg/kg Group 11 (18) 3.8 0.62 (18) . **0 . 0 0 0 0 0000 (18) 0.09 0.037 1 . 2 mg/kg (17) (17) (17) Group 12 1 . 6 mg/kg Group 13 3.5 0.32 GO) 3.8 0.62 0.003 0.0095 (1 0 ) 0 . 0 0 0 0 .0 0 0 0 0.11 0.032 (1 0 ) 0.09 0.017 2 mg/kg (4) (4) ________ w ________ 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* GO) 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 0.4 mg/kg 3.9 0.83 (2 0 ) 0 . 0 0 0 0 .0 0 0 0 (2 0 ) 0.08 0.043 (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) (1 0 ) (1 0 ) 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* (1 0 ) 7.4 1.54* (4) Levels of cholesterol, low density lipoproteins (LDL) and triglycerides in the blood were significantly increased (p < 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; p<0.05. Significantly different from control group; p<0.01. 000493 418-018.-PAGE III-24 Clinical Chemistry Values (mg/dL) 0 'PFOS plus Mevalonic Aicid Parameter Cholesterol Glucose LDL-Dir HDL-Dir Triglycerides Group 2 (Mevalonic) Group 3 1 . 6 mg/kg Group 4 2m /kg___ 115.4 7.6 (1 2 ) 157.0 23.7** (7) 0 . 0 0 0 .0 0 0 (0 ) 241.4 88.4 (1 2 ) 273.3 65.1 (7) 0 . 0 0 0 .0 0 0 (0 ) 35.3 7.5 (1 2 ) 50.8 20.0* (7) 0 . 0 0 0 .0 0 0 (0 ) 35.4 9.0 (1 2 ) 33.0 4.9 (7) 0 . 0 0 0 .0 0 0 (0 ) 193.2 69.9 (1 2 ) 302.3 62.7** (7) 0 . 0 0 0 .0 0 0 (0 ) Mevalonic Acid 335.3 229.2 (1 1 ) 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 Tj FreeT3 TSH Parameter ug/dL ng/dL pg/mL ng/mL Group 2 0.486 0.227 83.420 31.720* 0.497 0.490 0.430 0.212 (Mevalonic) (5) (5) (3) (2 ) Group 3 . **0 . 0 0 0 0 000 70.633 24.823 0 . 0 0 0 .0 0 0 0.680 0 . 0 0 0 1 . 6 mg/kg Group 4 2 mg/kg (3) (3) (0 ) (1 ) 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) 08) (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 000494 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 (Mevalonic) 3.6 0.69 (18) ' 0.003 0.0096 (18) 0.09 0.026 (18) 13.0 7.38 (18) Group 3 1 . 6 mg/kg 3.3 0.68 (1 0 ) 0.008 0.0253 GO) 0.09 0.037 GO) 13.6 5.38 GO) Group 4 2 mg/kg NO VALUES FOR THIS GROUP Levels of low (LDL) and high (HDL) density lipoproteins were significantly increased (><0.05 or /?<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 (><0.05) in the pups from dams that were treated with cholesterol and 2 mg/kg PFOS. Clinical Chemistry Values (mg/dL) of PFOS plus Cholesterol Parameter Cholesterol Glucose LDL-Dir HDL-Dir Group 5 (Cholesterol) 117.8 9.1 (1 2 ) 263.8 56.4 (1 2 ) 26.8 1 2 . 2 (1 2 ) 28.7 7.7 (1 2 ) Group 6 1 . 6 mg/kg 130.0 18.5 (8 ) 247.8 48.9 (8 ) 41.0 11.4* (8 ) 39.2 3.6** (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 ><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 Parameter Total T4 ug/dL Total T3 ng/dL Free T3 pg/mL Group 5 0.323 0.273" 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 Group 7 (3) 0 . 0 0 0 0 .0 0 0 (3) 25.900 0.000* (1) 0 . 0 0 0 0 .0 0 0 2 mg/kg (1 ) (1 ) (0 ) TSH ng/mL 1.380 0.000* (1 ) 0 . 0 0 0 0 .0 0 0 (1 ) 0 . 0 0 0 0 .0 0 0 (0 ) FreeT4 ng/dL 0.064 0.027 (5) . *0 . 0 0 0 0 000 (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 'Jver 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 2 mg/kg (1 1 ) 4.0 0.14 (4) (1 1 ) 0 . 0 0 0 0 .0 0 0 0 (4) (1 1 ) 0.10 0.037 (4) (1 1 ) 13.5 5.16 (4) * Significantly different from control group; p < 0 .0 5 . ** Significantly different from control group; ><0.01. a. Group 5 significantly different from Group 1; ><0.01 000495 418-018:PAGE ID-26 Liver triglyceride levels were significantly reduced (p<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 . 0 0 0 0 .0 0 0 0 (8 ) 0 . 0 0 0 0 .0 0 0 0 (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 Sera* Liver* 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 .0 0 245.2 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 .0 5 . ** Significantly different from control group; p<0.01. 000496 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. 000497 418-018:PAGE III-28 REFERENCES I. 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. F e d e r a l R e g is te r , 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). G o o d L a b o r a to ry P r a c tic e S ta n d a rd f o r S a fe ty S tu d ies on D ru g s, MHW Ordinance Number 21, March 26, 1997. 4. European Economic Community (1989). C o u n c il d e c isio n on 2 8 J u ly 1 9 8 9 on th e acceptance b y the E uropean E conom ic Com m unity o f an O E C D decision/recom - m en d a tio n o n c o m p lia n c e w ith p r in c ip le s o f g o o d la b o ra to ry p r a c tic e . 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 V ivo R e p ro d u c tiv e a n d M u ta g e n ic ity T ests. 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). E m b ry o a n d F e ta l D e v e lo p m e n ta l T o x icity (T e r a to lo g y ) C o n tro l D a ta in th e C h a rles R iv e r C rl:C D B R R at. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research.) 8. Institute of Laboratory Animal Resources (1996). G u id e f o r the C a re a n d U se o f L a b o r a to r y A n im a ls. 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. S ta tis tic a l M e th o d s, 6th Edition, Iowa State University Press, Ames, pp. 240-241. II. Sokal, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. B io m etry, W.H. Freeman and Co., San Francisco, pp. 370-371. 12. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. S ta tistic a l M e th o d s, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 000498 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. B io m etry, 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). N o n p a r a m e tr ic S ta tis tic s f o r th e B e h a v io ra l S c ie n c e s, McGraw- Hill, New York, pp. 96-104. 000499 APPENDIX A REPORT FIGURES ( ooa&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) BODY WEIGHTS Fo GENERATION FEMALE RATS Figure 1 - Groups 1,8 -1 3 450 GROUP 1 425 .CJ 400 GROUP 8 375 GROUP 9 350 0 H- X 325 0 LU 5 300 275 II GROUP 10 . x- GROUP 11 X- GROUP 12 418-018:PAGE A-l oC 250 o o( / I 225 CJ GROUP 13 *p < 0 .0 5 (from group 1) 200 I i 15 22 2*9 36 4^a i W 4 k t I i 10 11 12 13 14 1*5 16 17 18 l'9 20 21 ^ I k **p < 0 .0 1 (from group 1) DAY OF STUDY DAYOF GESTATION DAYOF 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 2 - Groups 2,3,4 450 - - -425 GROUP 2 400 375 350 0 h- 1 325 O 300 275 o 250 o o 225 ot n N 200 ,,w V X-S 'M I:i --a -- GROUP 3 GROUP 4 *p < 0 .0 5 (from group 2) **p < 0 .0 1 (from group 2) HU DAYOF GESTATION DAYOF LACTATION a. Last value recorded before cohabitaton. 418-018:PAGE A-2 PROTOCOL 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 5,6,7 450 425 400 375 350 0 S 325 LU 300 GROUP 5 ---a -- GROUP 6 GROUP 7 #p<0.05 (from group 1) *p < 0 .0 5 (from group 5) +*p<0.01 (from group 5) 275 418-018:PAGE A-3 000503 250 / 225 / 200 I 15 22 ^9 36 4^a DAY OF STUDY DAYOF GESTATION DAYOF LACTATION a. Last value recorded before cohabitaton. APPENDIX B REPORT TABLES 000504 PROTOCOL 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 18 VEHICLE CONTROL 0.4 9 0.8 10 11 12 1 1.2 1.6 PRECOHABITATION (DAY 1 OF STUDY TO THE DAY iOF COHABITATION): MAXIMUM POSSIBLE INCIDENCE 1176/ 28 840/ 20 840/ 20 840/ 20 840/ 20 1176/ 28 MORTALITY Q0000 0 LOCALIZED ALOPECIA: TOTAL UNDERSIDE LIMBS 61/ 4 14/ 2 47/ 2 29/ 2 0/ 0 29/ 2 0/ 0 0/ 0 0/ 0 14/ 2 0/ 0 14/ 2 26/ 1 0/ 0 26/ 1 32/ 1 0/ 0 32/ 1 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 0/ 0 0/ 0 0/ 0 5/ 1 0/ 0 5/ 1 28/ 2 3/ 1 25/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 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/ X 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 418-018:PAGE B-l o c/v W PROTOCOL. 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 21: CLINICAL OBSERVATIONS - 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 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/ 5 50/ 3 0/ 0 23/ 2 0/ 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/ I 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 CHROMORHINORRI1EA 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/ I 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). ooc o a 13 2 610/ 28 0 36/ 3 36/ 3 5/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 418-018:PAGE B-2 PROTOCOL 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 3) : CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 18 VEHICLE CONTROL 0.4 9 0.8 10 11 1 1.2 12 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 15/ 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 = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. 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/ 0 418-018:PAGE B- o 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 CHROMORHINORR1IEA 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/ 1 TIP OF TAIL MISSING 0/ 0 0/ 0 39/ 1 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 0/ 0 1/ 1 0/ 0 SCANT FECES 0/ 0 1/ 1 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. sosooo 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 7/ 5##a 2/ 2 11/ 7 CHROMORHINORRHEA 3/ 3##a 2/ 2 5/ 2 INCISORS: MISSING/BROKEN 3/ 3 4/ 2 3/ 2 PTOSIS 0/ 0 0/ 0 30/ 2 LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE NECK 33/ 3 10/ 1 23/ 2 0/ 0 22/ 3 9/ 1 5/ 2 12/ 1 5/ 1 5/ 1 0/ 0 0/ 0 CHROMODACRYORRHEA 1/ la 0/ 0 2/ 1 TIP OF TAIL MISSING 0/ 0 0/ 0 22/ 1 DEHYDRATION 0/ 0 0/ 0 5/ 1 RALES 0/ 0 0/ 0 5/ 1 EMACIATION 0/ 0 0/ 0 2/ 1 SCANT FECES 0/ 0 0/ 0 1/ 1 MOUTH : SCAB 3/ 1 4/ 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, tttt Significantly different from the Group 1 value (p<0.01). 6OS0bO 418-018:PAGE B-5 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 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 DECREASED MOTOR ACTIVITY SOFT OR LIQUID FECES 3/ 1 1/ 1 1/ la 1/ la 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 LACTATION : 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 ooc (A O PROTOCOL 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 47/ 6 44/ 5 11/ 2 3/ 1 13/ 2 13/ 2 0/ 0 0/ 0 12/ 1 12/ 1 0/ 0 0/ 0 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 17/ 1 0/ 0 17/ 1 8/ 1 8/ 1 0/ 0 2/ 1 2/ 1 0/ 0 CHROMORHINORRHEA 0/ 0 1/ 1 0/ 0 DECREASED MOTOR ACTIVITY 1/ la 0/ 0 0/ 0 PALE IN APPEARANCE 1/ la 0/ 0 0/ 0 COLD TO TOUCH 1/ la 0/ 0 0/ 0 SWOLLEN SNOUT 1/ 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 11562 wao moribund sacrificed on day 16 of study. 418-018:PAGE B-7 ooc Cft K4 H PROTOCOL 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 601/ 27 604/ 28 610/ 28 MORTALITY 00 0 LOCALIZED ALOPECIA: TOTAL NECK LIMBS UNDERSIDE BACK 77/ 5 22/ i 59/ 4 14/ 1 0/ 0 52/ 5 0/ 0 41/ 4 28/ 2 14/ i 54/ 5 0/ 0 40/ 4 22/ 1 2/ 1 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 32/ 2 0/ 0 32/ 2 17/ 2 16/ 2 7/ 1 9/ 2 9/ 2 0/ 0 CHROMODACRYORRH EA 3/ 1 2/ 1 8/ 2 CHROMORHINORRHEA 0/ 0 0/ 0 2/ 2 CHEST OR LEFT FORELIMB: ULCERATION 0/ 0 11/ 1 0/ 0 CHEST ; SCAB 0/ 0 4/ 1 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/ i 3/ i RED PERIVAGINAL SUBSTANCE 2/ 2 1/ 1 0/ 0 INCISORS: MISALIGNED 4/ 1 4/ I 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. 418-018:PAGE B-8 000S12 PROTOCOL 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 0 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 20 20 20 20 20 28 a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. 13 2 28 0 28 418-018:PAGE B-9 coo yi ** PROTOCOL 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 N1 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. 418-018:PAGE B-10 frtsooo PROTOCOL 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 QHOVd:8lO'8Ifr o 0 Cft 01 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) 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 1 1.2 12 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 MEAN+S.D. 14. 3 + 0.7 14.9 + 1.9 15.3 + 1.2 LIVER (%) MEAN+S.D. 3 . 4 + 0.2 3.7 + 0.5 3.7 + 0.3 DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION: RATS TESTED N 19 20 17 18 18 14 6 TERMINAL BODY WEIGHT MEAN+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 MEAN+S.D. 1 3 . 2 + 1.8 1 3 . 8 + 1.5 13.6 + 1.1 1 3 . 7 + 1.4 14.5 + 1.5 13.2 + 1.2 13.1 + 1.0 LIVER (l) MEAN+S.D. 4 . 1 + 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* ALL 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. 9TS000 418-018:PAGE B-I2 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 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 MEAN+S.D. 416.6 + 24.6 LIVER MEAN+S.D. 15.3 + 2.1 LIVER (t) MEAN+S.D. 3.7 + 0.4 DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION RATS TESTED N 18 TERMINAL BODY WEIGHT MEAN+S.D . 319.5 + 18.5 LIVER MEAN+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. 418-018:PAGE B-13 ooc c/1 H PROTOCOL 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 MEAN+S.D. 441.0 + 30.6 LIVER MEAN+S.D. 14.1 + 1 .3 LIVER (!) MEAN+S.D. 3.2 + 0.2 DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION: RATS TESTED N 17 TERMINAL BODY WEIGHT MEAN+S.D. 328.0 + 31.9 LIVER MEAN+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. 01) . 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL 8 403.0 + 38.3* 14.3 + 0.8 3.6 + 0.4* 8 387.5 + 3 1 . 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. 8TS000 418-018:PAGE B-14 PROTOCOL 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 lGROUP DOSAGE 1MG/KG PFOS 1 VEHICLE 1CONTROL 8 0 .4 RATS TESTED N 28 20 BODY WEIGHT (G) DAY 1 MEAN+S.D. 215.4 + 10.0 215.0 + 12.7 DAY 8 MEAN+S.D. 229.8 + 12.1 236.2 + 11.0 DAY 15 MEAN+S.D. 238.5 + 14.8 247.5 + 13.8 DAY 22 MEAN+S.D. 249.9 + 17.6 257.4 + 12.9 DAY 29 MEAN+S.D . 255.3 + 17.5 262.4 + 15.0 DAY 36 MEAN+S.D. 262.6 + 18.5 271.4 + 16.3 DAY 42a MEAN+S.D. 265.6 + 2l .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 + 17.5 418-018:PAGE B-15 000519 PROTOCOL 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 MEAN+S.D. 216.2 + 11.3 215.6 + 12.1 DAY 8 DAY 15 DAY 22 DAY 29 DAY 36 DAY 42b MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D . MEAN+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 l 28) a 258.1 + 19.5 [ 28) a 261.3 + 23.1 I 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. La3t 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 91-9 3 0 Vd:810-8117 U00520 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 4 (PAGE 3): BODY WEIGHTS - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 1 * CHOLESTEROL RATS TESTED N 28 28 BODY WEIGHT (G) DAY i MEAN+S.D. 214.8 + 10.1 215.7 + 10.5 DAY 8 MEAN+S.D. 232.8 + 11.4 231.5 + 10.8 DAY 15 MEAN+S.D . 242.8 + 14.3 240.8 + 12.4 DAY 22 DAY 29 DAY 36 DAY 42b MEAN+S.D . MEAN+S.D. MEAN+S.D. MEAN+S.D. 254.2 + 15.4 ( 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 l I = 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 (p<0.01). 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 000921 PROTOCOL 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 i VEHICLE CONTROL 8 0 .4 9 0 .8 RATS TESTED N 28 20 20 BODY WEIGHT CHANGE (G) DAYS 1 - 8 MEAN+S.D . *14.5 + 8.6 +21.2 + 6.6** +17.7 + 6.4 DAYS 8 - 15 MEAN+S.D . +8.7 + 5.2 +11.2 + 7.5 +11.6 + 5.2 DAYS 15 - 22 MEAN+S.D. +9.4 + 6.9 +10.0 + 6.8 +7.8 + 4.9 DAYS 22 - 29 MEAN+S.D. +7.4 + 5.9 +4.9 + 5.9 +4.6 + 6.8 DAYS 29 - 36 MEAN+S.D . +7.3 + 4.7 +9.0 + 5.7 +7.6 + 5.2 DAYS 36 - 42a MEAN+S.D . +3.0 + 8.4 +4.7 + 7.6 +1.6 + 9.1 DAYS 1 - 42a MEAN+S.D. +50.2 + 17.3 +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 (p<0.05). (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* +4.2 + 4.2* -0.1 + 6.0 +1.5 + 6.8 +44,5 + 10.4 +39.2 + 16.4* 418-018:PAGE B-18 000522 PROTOCOL 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 418-018:PAGE B-19 CO + o CO 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 BODY WEIGHT CHANGE (G) DAYS 1 - 8 MEAN+S.D. +13.4 + +15.9 + 7.6 DAYS 8 - 15 MEAN+S.D. +10.6 + 5.7 +8.8 + 6.6 DAYS 15 - 22 MEAN+S.D. +8.9 + 5.5 +9.0 + 5.4 DAYS 22 - 29 MEAN+S.D. +8.3 + 6.5 +4 .1 + 4.7** DAYS 29 - 36 MEAN+S.D. +8.0 + 5.9 +4 .5 + 4.2** DAYS 36 - 42b MEAN+S.D. +2.1 + 6.9 +3.2 + 6.5 DAYS 1 - 42b MEAN+S.D. +51.4 + 13.7 +45.6 + 16.6 DAYS = DAYS OF STUDY a. Excludes values Cor 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 + 3.7* +3.3 + 5.0 +50.8 + 10.6 CZS0OO PROTOCOL 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 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL RATS TESTED N 2B 28 BODY WEIGHT CHANGE (G) DAYS 1 - 8 MEAN+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 MEAN+S.D. MEAN+S.D. MEAN+S.D . MEAN+S.D . MEAN+S.D . +10.7 + 4.8 [ 27] a +10.3 + 7.0 ( 27) a +7.7 + 7.8 l 27] a +3.4 + 7.4 1 27] a +60.7 + 16.8# ( 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 [ ) = 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. tt Significantlydifferent from the Group 1 value (p<0.05). * Significantly different from the Group 5 value (p<0.05). ** Significantlydifferent from the Group 5 value (p<0.01). 7 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 +42.5 + 14.0** 418-018:PAGE B-20 ooc ctno PROTOCOL 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 MEAN+S.D. 275.0 + 21.6 281.6 + 17.0 274.0 + 22.1 DAY 1 MEANfS.D. 283.3 + 20.4 291.2 + 18.5 281.1 + 23.3 DAY 2 MEAN+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 MEAN+S.D. 293.6 + 19.9 304.0 + 19.8 290.4 + 23.7 DAY 6 MEAN+S.D. 295.4 + 20.2 306.2 + 20.1 289.8 + 22.0 DAY 7 MEAN+S.D. 298.9 + 20.0 308.4 + 19.2 292.5 + 21.7 DAY 8 MEAN+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** szsooo 418-018:PAGE B-21 PROTOCOL 418-018: OHE 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 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 ii MEAN+S.D. 313.8 + 21.1 324.2 + 19.5 308.6 + 23.2 DAY 12 MEAN+S.D. 318.2 + 22.3 329.8 + 19.2 312.9 + 24.7 DAY 13 MEAN+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 MEAN+S.D. 333.9 + 23.0 347.1 + 20.9 332.4 + 25.3 DAY 16 MEAN+S.D. 343 .1 + 24 .1 357.0 + 22.6 343.3 + 26.0 DAY 17 DAY 18 DAY 19 DAY 20 DAY 21 MEAN+ S .D . MEAN+S.D . MEAN+S.D. MEAN+S.D. MEAN+S.D. 356.0 + 26.6 370.1 + 28.6 381.9 + 29.9 396.2 _+ 31.3 410.6 + 31.1 370.4 + 23.0 383.6 + 23.9 395.8 + 24 .4 410.3 + 26.0 421.2 + 28.9 ( 18] a 357.0 + 26.4 371.2 + 26.1 384 .6 + 28.0 395.6 + 26.2 408.5 + 25.3 ( 12]1a DAY = DAY OF GESTATION I 1 = 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). 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 + 22.2 1 12] a 11 1 .2 20 18 302.2 + 19.9 306.8 + 22.5 309.3 1 19.7 316.2 + 19.4 322.4 + 21.2 333.3 + 21.3 347.9 + 22.5 362.7 + 23.7 373.2 + 25.3 387.1 + 27.0 394.5 + 31.7 i 15] a 12 13 i .6 2 28 28 25 27 296.4 + 18.4** 291.2 + 19.4** 299.1 + 19.2** 294.4 + 18.8** 304.3 + 19.4** 299.2 + 18.4** 310.4 + 18.2* 302.9 + 19.0** 319.1 + 19.0* 313.3 + 19.4** 328.7 + 19.6* 323.4 + 19.3** 343.4 + 20.6 358.5 + 20.3 373.0 + 22 .8 385.1 + 25.0 398.3 + 26.1 [ 20] a 337.3 + 20.0** 352.1 + 21.0* 365.6 + 21.5* 380.0 + 23.3 397.5 + 30.2 I 21] a 9ZS000 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-6295.2S) 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 MEAN+S.D. 275.3 + 19.0 DAY 1 MEAN+S.D. 283.0 + 18.8 DAY 2 MEAN+S.D. 286.3 + 18.9 DAY 3 MEAN+S.D . 287.0 + 18.9 DAY 4 MEAN+S.D . 289.0 + 18.9 DAY 5 MEAN+S.D. 290.8 + 19.3 DAY 6 MEAN+S.D . 292.9 + 19.0 DAY 7 MEAN+S.D . 296.0 + 17.9 DAY 8 MEAN+S.D . 299.0 + 18.2 DAY 9 MEAN+S.D . 302.6 + 19.4 DAY 10 MEAN+S.D . 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 000527 PROTOCOL 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 4): MATERNAL BODY WEIGHTS - 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 (G) DAY ii DAY 12 MEAN+S.D. MEAN+S.D. 311.7 + 18.5 313.5 + 18.3 294.4 + 24.9** 299.0 + 25.2* 300.7 1 20.5 303.3 + 21.0 DAY 13 DAY 14 DAY 15 DAY 16 DAY 17 DAY 18 DAY 19 DAY 20 DAY 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 318.3 + 18.6 I 26) b 322.8 + 18.8 [ 26] b 330.6 + 18.8 ( 26] b 339.4 + 18.8 1 26] b 351.0 + 2 0 . 7 [ 26] b 363.4 + 20.8 [ 26] b 379.4 + 24.9 ( 26] b 390.9 + 24.7 ( 26] b 403.5 + 2 4 . 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 l 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 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). 12 of gestation (death was attributed to an intubation 8ZS000 418-018:PAGE B-24 PROTOCOL 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 MEAN+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 MEAN+S.D. 299.4 + 22.6 272.6 + 18.5** DAY 4 MEAN+S.D. 302.3 + 21.4 274.2 + 18.3** DAY 5 MEAN+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 MEAN+S.D. 309.8 + 24.0 281.5 + 20.5** DAY 8 MEAN+S.D . 315.1 + 25.0# 283.8 + 21.0** DAY 9 MEAN+S.D . 317.7 + 24.6# 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. II Significantly different from the Group 1 value (p<0.05). ** 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** 6ZS00 418-018:PAGE B-25 PROTOCOL 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 67 1.6 + CHOLESTEROL RATS TESTED N 27a 28 PREGNANT N 25 28 MATERNAL BODY WEIGHT (G) DAY 11 MEAN+S.D . 329.0 + 24.2# 295.8 + 21.4" DAY 12 MEAN+S.D . 332.9 + 25.1# 300.8 + 22.6" DAY 13 MEAN+S.D . 335.7 + 26.0# 305.6 22.1** DAY 14 MEAN+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 MEAN+S.D . 357.3 + 26.8# 329.0 + 26.4** DAY 17 MEAN+S.D . 370.2 + 26.2# 342.4 + 26.8" DAY 18 MEAN+S.D . 383.2 + 27.0 357.2 + 29.2** DAY 19 MEAN+S.D . 397.1 + 28.6 370.1 + 31.7" DAY 20 MEAN+S.D . 410.3 + 29.6 383.6 + 32.6** DAY 21 MEAN+S.D . 428.0 + 31.1# l 24]b 392.0 + 31.6** [ 23] b 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. (I Significantly different from the Group 1 value (p<0.05) . ** Significantly different from the Group 5 value (p<0.01). 2 + CHOLESTEROL 28 27 292.8 + 18.7** 296.3 + 20.4** 300.8 + 21.5** 305.4 + 21.0** 314.4 1 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** t 19] b ocsooo 418-018:PAGE B-26 PROTOCOL 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 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 1 1.2 12 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 + 2 6 . 8 + 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 l 18] a +86.2 + 7.6 l 12] a +133.4+13.7 ( 12 )a +78.4 + 25.6 [ 12] a +132.6 + 29.6 [ 12) a +79.9 + 16.6 t 15] a +128.3 + 23.5 t 15] a +87.2 + 15.3 l 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 l value ** Significantly different from the Group 1 value (p<0.05). (p<0.01). 418-018:PAGE B-27 000531 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 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 MEAN+S.D . +20.7 + 6.1 +13.4 + 7.4** +12.7 + 7.8** DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 2 1 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 + 18.3 [ 25] c +27.6 + 8.2 +88.8 + 2 3 . 8 [ 19 Jc +130.1 + 3 0 . 1 ( 19] c DAYS = DAYS OF GESTATION ( J = 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 (death was attributed to an intubation accident). c. Excludes values for dams that delivered. ^ ** Significantly different from the Group 2 value (p<0.01). 418-018:PAGE B-28 ccc ui U K 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 7 (PAGE 3): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED N 5 CHOLESTEROL CONTROL 27a 6 1.6 + CHOLESTEROL 28 PREGNANT N 25 28 MATERNAL BODY WEIGHT CHANGE (G ) DAYS 0 - 7 MEAN+S.D. +25.0 + 10.4 +16.4 + 8.9** DAYS 7 - 14 MEAN+S.D. +30.9 + 6.9 +28.6 + 8.2 DAYS 14 - 21 DAYS 0 - 2 1 MEANtS.D. MEAN+S.D. +86.5 + 14.9 [ 241b +142.8 + 19.8 1 24)b +86.4 +18 . 5 ( 23 ]b +129.7 + 22.8* 1 23)b 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 (pc0.05). ** 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 1 191 b +128.5 + 15.4* [ 191b 418-018.PAGE B-29 000S33 PROTOCOL 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 1CONTROL 8 0.4 9 0.8 10 ii 1 1.2 12 1.6 RATS TESTED N 19 20 20 20 20 20 PREGNANT N 19 20 17 19 18 17 DELIVERED A LITTER N 19 20 17 19 18 17 BODY WEIGHT (G) DAY I MEAN+S.D. 3 d .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 MEAN4-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 B-30 000534 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 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 MEAN+S.D . 299.3 +24 . 8 301.4 + 28.6 300.8 + 20.0 DAY 5 MEAN+S.D. 319.5 + 18.5 315.8 + 23.4 ( 12] b 297.0 + 14.1 ( 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- c o Cfl Ci tfl 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 1) : 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 MEAN+S.D . 312.4 + 24.3 294.8 + 24.8` 295.4 + 22.6* DAY 5 MEAN+S.D. 328.0 + 31.9 303.9 + 19.0* [ 13] b 280.8 + 15.6** [ 7]b DAY = DAY OF LACTATION I 1 = 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 (p<0.05). ** Significantly different from the Group 5 value (p<0.01). 418-018:PAGE B-32 c o V0 I CJ 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 9 (PAGE 1) : BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL 8 0.4 9 0.8 10 ii 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 ANALYS1ES N 19 20 17 18a 18 14a 6a BODY WEIGHT CHANGE (G) DAYS I - 5 MEAN+S.D. +17.4 + 1 1 . 1 +15.7 + 15.4 +4.3 + 25.8* +4.0 + 10.S* +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 1 value (p<0.05). ** Significantly different from the Group 1 value (p<0.01). 418-018:PAGE B-33 coo C/l u 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 - S MEAN +S .D . +20.2 + IS.3 +7.7 + 15.5* +1.7 + 11.2 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). 418-018:PAGE B-34 ooo Cfl Q 00 PROTOCOL 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 prior to day 5 of lactation. * Significantly different from the Group 5 value (p<0.05). 6CS000 418-018:PAGE B-35 PROTOCOL 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 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE 1CONTROL 8 0.4 RATS TESTED N 28 20 FEED CONSUMPTION (G/DAY) DAYS 1 - 8 DAYS 8 - 15 MEAN+S.D. MEAN+S.D. 19.6 + 3.0 l 26) a 19.0 + 1.8 19.6 + 1.9 ( 19) a 19.0 + 3.6 DAYS 15 - 22 DAYS 22 - 29 MEAN+S.D. 10.9 + 2 .3 MEAN+S.D. 18.8 + 2.0 1 9 . 4 + 1.8 ( 19) a 1 9 . 4 + 2.0 DAYS 29 - 36 MEAN+S.D. 18.8 + 2 .1 DAYS 36 - 42b MEAN+S.D. 18.3 + 2 .1 20.2 + 2.2* [ 17] a 19.4 + 2.1 DAYS 1 - 42b MEAN+S.D. 18.9 + 1.8 19.6 + 1.0 9 0.8 20 19.1 + 1.9 i 19) a 1 8 . 9 + 1.5 ( 19] a 19.2 + 1.6 [ 18] a 19.0 + 2.5 t 19) a 1 8 . 3 + 2.1 [ 18] a 18.2 + 2.6 1 9 . 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 t 19] a 19.4 + 1.8 [ 19] a 19.2 + 1.5 1 8 . 7 + 1.8 19.1 + 1.4 [ 17] a 18.8 + 1.7 ( 19] a 18.3 + 1.7 [ 19] a 17.4 + 2.2 l 19] a 18.6 + 1.3 [ 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 I 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 17.8 + 1.3* [ 27] a 418-018:PAGE B-36 000540 PROTOCOL 4 18-018 : 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 2 I : ABSOLUTE FEED CONSUMPTION VALUES (G/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/DAY) DAYS 1 - 8 DAYS 8 - 15 MEAN+S.D. MEAN+S.D. 19.9 + 1.4 [ 26) b 20.1 + 2.2 19.2 + 1.9 19.0 + 3.4 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. 20.1 + 2.2 [ 27] b 20.0 + 1.9 19.4 + 2.2 1 26] b 18.8 + 2.1 19.8 + 1.7 18.8 + 2.5 I 27] b 18.0 + 2.4** ( 27] b 18.4 + 2.2 17.5 + 2.3* [ 271b 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 1 9 . 5 + 1.4 [ 27] b 18.8 + 2.1 19.0 + 1.9 t 27] b 18.4 + 1.7** ( 26] b 18.3 + 2.5 [ 26] b 17.4 + 2.3* 18.7 + 1.5* 418-018:PAGE B-37 ooc Cfl PROTOCOL 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 67 1.6 + CHOLESTEROL RATS TESTED N 28 28 FEED CONSUMPTION (G/DAY) DAYS 1 - 8 MEAN+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 MEAN+S.D . MEAN+S.D . MEAN+S.D. MEAN+S.D . MEAN+S.D . MEAN+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 1 9 . 7 + 2. 1# 1 27] b 19.7 + 1. 7 [ 27] b 18.6 + 1.3 18.8 + 2.1 17.9 + 1.5** 18.3 + 1.4** [ :23] a 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. # Significantly differentfrom theGroup1 value(p<0.05). Htt Significantly differentfrom theGroup1 value(p<0.01). ** Significantlydifferent from the Group 5 value (p<0.01). 2 + CHOLESTEROL 28 19.0 + 1.5 18.9 + 1.9 18.9 + 2.1 1 27] a 17 7 1 1.9** 18.2 + 2.4** [ 25] a 17.3 + 1.6** 18.4 + 1.6** 418-018:PAGE B-: 000542 OJ 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 1 VEHICLE CONTROL 8 0.4 RATS TESTED N 28 20 FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 DAYS 8 - 1 5 MEAN+S.D. MEAN+S.D. 88.2 + 14.0 ( 26!1a 80.9 + 4 .9 8 7 . 0 + 8.3 ( 19]a 78.4 + 14.4 DAYS 15 - 22 DAYS 2 2 - 2 9 MEAN+S.D. 77.7 + 6.3 MEAN+S.D. 74.9 + 5.3 77.1 + 6.8 ( 19) a 7 4 . 6 + 6.2 DAYS 29 - 36 MEAN+S.D. 72.6 + 6.4 DAYS 36 - 42b MEAN+S.D. 69.1 + 5.4 76.6 + 6.9 ( 17) a 7 0 . 7 + 5.3 DAYS 1 - 42b MEAN+S.D. 77.2 + 4 .6 77.6 + 5.8 9 0.8 20 85.3 + 7.8 [ 19) a 79.4 + 4.9 [ 19) a 77.4 + 3.8 ! 18) a 74.5 + 7.4 [ 19) a 70.6 + 5.2 [ 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 ( 19] a 82.7 + 8.8* 77.4 + 6.9 75.1 + 7.8 [ 19) a 68.8 + 6.7 ( 19] a 78.8 + 6.3 [ 19] a 85.8 + 6.6 78.6 + 4.9 77.9 + 3.4 [ 17 ]a 74.1 + 6.5 I 19) a 70.2 + 5.9 [ 19) a 65.8 + 6.7 [ 19] a 75.4 + 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 73.2 + 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 1 27) a 71.1 + 6.0* 68.2 + 4.9* [ 26] a 63.9 + 4.3** ( 27] a 7 3 . 4 + 3.8* [ 27] a 418-018:PAGE B-39 000543 PROTOCOL 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. MEAN+S.D. 89.5 + 6.9 [ 26) b 85.6 + 6.6ft# 85.8 + 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 + 7.0ft l 27] b 78.8 + 6.2ft 7 4 . 6 + 6.5 ( 26) b 70.4 + 6.8 80.3 + 4.5# 76.9 + 7.9** I 27) b 71.6 + 6.2** I 27] b 71.9 + 6.3 67.3 + 5.6 [ 27) b 75.7 + 5.3** [ 27] b DAYS = DAYS OF STUDY I ) = 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. ft Significantly different from the Group 1 value (p<0.05). ftft Significantly different from the Group 1 value (p<0.01). * 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 86.8 + 5.6 t 27] b 78.9 + 7.7** 76.9 + 7.5** [ 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 oc o Cft PROTOCOL 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 S CHOLESTEROL CONTROL 6 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 D A Y S 8 - 15 DAYS 15 - 22 DAYS 2 2 - 29 D A Y S 29 - 36 DAYS 36 - 42c DAYS 1 - 42c MEAN+S.D. M E A N + S .D . M E A N + S .D . M E A N + S .D . MEAN+S-D. MEAN+S.D. 80.8 + 5.2 ( 27]a 80.2 + 6.9 1 27] b 76.2 + 6.3 t 27] b 77.9 + 8.0## [ 25] a ,b 71.9 + 6.5 [ 27] b 78.6 + 4.7 l 27] b 79.0 + 4.0 76.7 + 6.9 71.4 + 3.9** 71.5 + 3.6** [ ;23] ai 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. (ttt 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). 7 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** l 25] a 67.6 + 6.5** 75.5 + 5.7* 418-018:PAGE B-41 000545 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 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 MEAN+S.D. 22.9 + 2.2 DAYS 14 - 21 MEAN+S.D. 24.3 + 2.2 DAYS 0 - 2 1 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 [ 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 a3 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** 19.0 + 2.1** 18.6 + 2.2** ( 26] a 22.0 + 2.1 21.2 + 2.7 21.0 + 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 418-018:PAGE B-42 c o C/t PROTOCOL 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 MEAN+S.D. 21.6 + 1.8 18.2 + 2.3*+ 19.1 + 2 . 1 ** 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 1 26] b 23.0 + 1.8 [ 26] b 20.9 + 1.9** 25.0 + 2.4 ( 22] d 21.5 + 2.0** I 22] d 20.8 + 2.0** [ 25] C 23.8 + 3.6 [ 24] d 2 1 . 5 + 2.2** [ 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 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). 12 of gestation (death was attributed to an intubation 418-018`.PAGE B-43 000S47 PROTOCOL 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. 22.9 + 2.9 ( 23) b 2 4 . 3 + 2.7 [ 24 ]b 2 5 . 4 + 2.9 ( 23) c 2 4 . 3 + 2.6 [ 23) C 18.9 + 2.2** 21.2 + 2.1** t 27) b 2 4 . 6 + 2.1 ( 26] C 21.5 + 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 dam3 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 000548 PROTOCOL 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 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/KG/DAY) DAYS 0 - 7 MEAN+S.D. 77.2 + 5.7 76.3 + 5.0 72.2 + 5.4* 7 2 . 3 + 7.2* DAYS 7 - 14 DAYS 1 4 - 2 1 DAYS 0 - 2 1 MEAN+S.D. 7 3 . 4 + 4.2 MEAN+S.D. 6 6 . 8 + 5.4 MEAN+S.D. 7 1 . 9 + 4.0 72.2 + 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 I 11]a,b 70.6 + 8.3 68.1 + 9.0 [ 12] b 71.2 + 6.9 ! 12] b 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 70.3 + 5.2** 73.7 + 6.8 68.6 + 5.4** 71.8 + 6.3 69.0 + 6.6** [ 26] a 72.7 + 11.4 6 8 . 9 + 5.8 1 15] b 70.4 + 4.5 [ IS] b 72.1 + 5.8* I 20] b 70.8 + 3.6 1 20] b 70.7 + 7.5 [ 20] a, b 71.0 + 5.6 [ 20] a ,b 6*S000 418-018:PAGE B-45 PROTOCOL 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 MEAN+S.D. 75.4 + 6.1 66.3 + 6.4** 6 8 . 4 + 8.2** DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. 74.9 + 6.0 1 24)b,c 67.9 + 4.8 1 25:1b, c 72.5 + 4.0 [ 25)b,c 71.5 + 3.4* 71.8 + 5.9 [ 21] d 7 0 . 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 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). 12 of gestation (death was attributed to an intubation ossoor 418-018 :PAGE B-46 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 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. 76.7 + 7.8 ( 23] b 74.8 + 6.7 [ 24 )b 6 6 . 9 + 6.4 ( 23) C 72.2 + 5.7 ( 23) C 69.1 + 6.0++ 7 1 . 8 + 5.1 l 27) b 7 1 . 2 + 4.4+ [ 23) C 70.6 + 3.5 ( 231 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<0.05). ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 28 27 67.5 + 7.2** 71.8 + 5.3 t 26] b 7 2 . 6 + 6.0+* ( 19] C 70.4 + 5.3 [ 19] C 418-018:PAGE B-47 c Cfl PROTOCOL 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 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 IB 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/DAY) DAYS 1 - S MEAN+S.D. 28.8 + 4.7 ( 18Jb 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). ZS S 0 6 & 418-018:PAGE B-48 PROTOCOL 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 1 - 5 MEAN+S.D. 28.4 + 5.3 20.2 + 5.6** ( lllc 13.9 + 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 PROTOCOL 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 31 : 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 17 20 19 INCLUDED IN ANALYSES N 17 13b 7b FEED CONSUMPTION (G/DAY) DAYS 1 - 5 MEAN+S.D. 3 1 . 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 r 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 15 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED PREGNANT DELIVERED A LITTER N N N 1 VEHICLE CONTROL 19 19 19 a 0.4 20 20 20 9 0.8 20 17 17 10 11 1 1.2 12 1.6 20 20 20 19 18 17 19 18 17 13 2 20 19 19 INCLUDED IN ANALYSES N 19 20 17 18a 18 14a 6a FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 5 MEAN+S.D. 92.8 + 13.7 [ 18] b 92.3 + 32.1 77.5 + 15.0* 83.0 + 29.G* 78.5 + 15.3* 65.1 + 18.9** 60.7 + 26.6 DAYS - DAYS OF LACTATION l ] = NUMBER OF VALUES AVERAGED a. Excludes values for dam3 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<0.05). ** Significantly different from the Group 1 value (p<0.01). sssooo 418-018:PAGE 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 IS (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** ( lllc 46.9 + 6.2** DAYS = DAYS OF LACTATION I ] = 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). 9SS000 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 4 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 prior to day 5 of lactation. ** Significantly different from the Group 5 value (p<0.01). 418-018:PAGE B-53 (fl c/v nI PROTOCOL 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 1 1.2 12 1.6 13 2 RATS IN COHABITATION N 28 20 20 20 20 28 28 DAYS IN COHABITATION a MEAN+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 20/ 20 17/ 19 19/ 20 18/ 19 25/ 27 27/ 28 (%) ( 96.4) (100.0) ( 89.5) ( 95.0) ( 94.7) ( 92.6) ( 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 (%) K 3.6) 0( 00.0) 0( 00.0) 1< 5.0) 0( 00.0) 1( 3.7) 1( 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 rats 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 ooc Cfl Cft 00 PROTOCOL 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 28a DAYS IN COHABITATION b MEAN_fS .D . 2.8 + 2.2 2.0 1 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) 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) K 3.6) 27/ 28 ( 96.4) SSQ D O 418-018:PAGE B-55 PROTOCOL 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 MEAN+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 N (%) 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) 1( 3.6) 27/ 28 ( 96.4) O9S00) 418-018:PAGE B-56 PROTOCOL 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 (%1 RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N CORPORA LUTEA MEAN+S.D . IMPLANTATIONS MEAN+S.D . LITTER SIZES MEAN+S.D . 1 VEHICLE CONTROL 9 8( 88.9) 8 1 6 . 1 + 1.6 1 4 . 9 + 1.2 1 3 . 5 + 1.3 12 1.6 8 8(100.0) 8 15.4 + 2.7 14.0 + 2.9 13.0 + 2.8 13 2 8 8(100.0) 8 18.9 + 5.1 15.2 + 1.8 14.9 + 1.7 T9S0bO \ 418-018:PAGE B-57 LIVE FETUSES N MEAN+S.D . 108 1 3 . 5 + 1.3 DEAD FETUSES N MEAN+S.D. 0 0.0 + 0.0 RESORPTIONS MEAN+S.D . 1.4 + 1.1 EARLY RESORPTIONS N MEAN+S.D . 11 1 .4 1.1 LATE RESORPTIONS N MEAN+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 418-010: 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 MEAN+S.D . LITTER SIZES MEAN+S.D . 2 MEVALONIC ACID CONTROL 8 8(100.0) 3 1.6 + MEVALONIC ACID 8 8(100.0) 8 19.1 + 3.3 15.4 + 1.1 13.6 + 2.7 8 17.5 + 4.1 15.5 + 2.4 13.5 + 3.1 4 2 + MEVALONIC ACID 8 8(100.0) 8 18.2 + 2.6 16.1 + 1.0 13.6 + 2.4 LIVE FETUSES N MEAN+S.D. DEAD FETUSES N RESORPTIONS MEAN+S.D . EARLY RESORPTIONS N MEAN+S.D . LATE RESORPTIONS N MEAN+S.D . DAMS WITH ANY RESORPTIONS N (%) c DAMS WITH ALL CONCEPTUSES DEAD OR RESORBED i . DAMS WITH VIABLE FETUSES o PLACENTAE 10 APPEARED NORMAL N(%) N <%) N(t) 109 1 3 . 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 1 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 PROTOCOL 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 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 MEAN+S.D . IMPLANTATIONS MEAN+S.D . LITTER SIZES MEAN+S.D. 5 CHOLESTEROL CONTROL 8 8(100.0) 8 17.8 + 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) 8 18.1 + 6.8 13.2 + 4.4 11.5 + 3.9 LIVE FETUSES N MEAN+S.D. DEAD FETUSES N RESORPTIONS MEAN+S.D . EARLY RESORPTIONS N MEAN+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 1 4 . 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 + 3.9 0 1.8 + 1.4 14 1.8 + 1.4 0 6{ 75.0) 0( 0.0) 8(100.0) 8(100.0) 418-018:PAGE B-59 000S63 PROTOCOL 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 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE iCONTROL LITTERS WITH ONE OR MORE LIVE FETUSES N 8 IMPLANTATIONS MEANj_S .D . 14.9 + 1.2 LIVE FETUSES N MEAN+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 MEAN+S.D . 9. ! + 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* +9S000 418-018:PAGE B-60 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 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 MEANfS.D. t 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 + 17.3 418-018:PAGE B-i 00Q565 Os PROTOCOL 418-018: OHE 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 MEAN+S.D . 5 CHOLESTEROL CONTROL e 15.9 + 2.2 6 1.6 + CHOLESTEROL 8 14.6 + 4.2 7 2 + CHOLESTEROL 8 13.2 + 4.4 LIVE FETUSES N MEAN+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 MEAN+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 000566 PROTOCOL 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 18 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 MEAN+S.D. 22.9 + 0.3 22.6 + 0.5 9 10 11 12 0.8 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 N 279 232 257 303 275 243 274 PER DELIVERED LITTER MEAN+S.D . 14.7 + 2.3 1 6 . 2 + 1.8 15.1 + 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) 1( 5.3)* K 5.6)* 1( 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 % 100.0 100.0 100.0 100.0 100.0 100.0 100.0 N/N 19/ 19 20/ 20 17/ 17 19/ 19 18/ 18 17/ 17 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 000567 PROTOCOL 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 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 MEAN+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) 22.2 + 0.4* 4 2 + MEVALONIC ACID 20 19( 95.0) 19(100.0) 22.0 + 0.4** IMPLANTATION SITES N PER DELIVERED LITTER MEAN+S.D . 263 1 4 . 6 + 2.0 251 13.9 + 2.2 289 15.2 + 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 for rats that died or were moribund sacrificed prior to lactation. b. Calculated as the time (in days) elapsed between confirmed mating (arbitrarily defined aa 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 coo 00 PROTOCOL 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 Nil) DELIVERED LITTERS N (%) DURATION OF GESTATION b MEAN+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) 2 2 . 0 + 0.4*+ 1 2 + CHOLESTEROL 20 19( 95.0) 19(100.0) 22.3 + 0.4** IMPLANTATION SITES N PER DELIVERED LITTER MEAN+S.D . 241c 15.1 + 1.4 I 16) C 288 1 4 . 4 + 1.7 291 15.3 + 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) K 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 o C/1 c* PROTOCOL 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 18 VEHICLE CONTROL 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 MEAN+S.D. 265 300 247 287 253 231 252 13.9 + 2.6 15.0 + 2.3 1 4 . 5 + 2.3 15.1 + 2.3 14.0 + 2.9 13.6 + 2.8 1 3 . 3 + 2.5 LIVEBORN MEAN+S.D. 13.7 + 2.7 1 4 . 6 + 2.4 1 4 . 4 + 2.4 15.0 + 2.3 1 4 . 0 + 2.9 1 3 . 4 + 2.9 1 3 . 2 + 2.5 N (%) 260( 98 .1) 291( 97.0) 24 5 ( 99.2) 285 ( 99.3) 252( 99.6) 227( 98.3) 251( 99.6) STILLBORN MEAN+S.D. N (%) 0.3 + 0.6 5< 1 .9) 0.4 + 0.6 9( 3.0) 0.1 + 0.3 2( 0.8) 0 . 1 + 0.4 2( 0.7) 0.0 + 0.2 1( 0.4) 0 . 2 + 1.0 4( 1.7) 0.0 + 0.2 K 0.4) UNKNOWN VITAL STATUS N 0 00 0 0 0 0 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2- 5 N/N (%) 1/260 ( 0.4) 3/2911 1.0) 1/245 ( 0.4) 4/285 ( 1.4) 6/252 ( 2.4) 13/227( 5.7) 56/251(22.3)" N/NU) 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)** VIABILITY INDEX a % 97.3 97.6 93.1 88.8 81.7 49.3 17.1 N/N 253/260 284/291 228/245 253/285 206/252 112/227** 43/251** D AY(S) = DA 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). sooo 418-018:PAGE B-66 o PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) T A B L E 20 (PAGE 2): LITTER O B S E R VATIONS (NATURALLY D E L I V E R E D PUPS) - SUMMARY - FI G E N E R A T I O N 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 DAY 2 DAY 3 MEAN+S.D. MEAN+S.D. MEAN+S.D. 13.7 + 2.7 14.6 + 2.4 14.4 + 2.4 15.0 + 2.3 14.0 + 2.9 1 3 . 4 + 2.9 13.2 + 2.5 13 .5 + 2.5 14 .4 + 2.4 13.9 + 2.2 13.8 + 3.0 12.9 + 3.2 8.0 + 5.7* 3.4 + 4.4** 13.4 + 2.5 14 .3 + 2.4 13.6 + 1.8 13.5 + 4.0 12.1 + 3.3 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 MEAN+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** PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY lb MEAN+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 46.6 + 15.2 t 18] C 46.3 + 15.1 [ 18] C 46.5 + 15.3 [ 18] C 49.6 + 14.6 48.5 + 15.0 48.5 + 14.6 47.7 + 15.9 46.0 + 21.3 [ 14 ]C 46.1 + 27.4 [ 14 ]C 46.1 + 26.7 ( 14 ]C 39.1 + 25.1 [ 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 ( 5]C DAY = DAY POSTPARTUM ( ) = NUMBER OF VALUES AVERAGED a. A v e r a g e n u m b e r of live pups per litter, i n c l u d i n g litters w i t h n o s u r v i v i n g pups. b. I n c l u d e s p u p s b o r n alive, found d e a d d a y 1 postpartum. c. E x c l u d e s v a l u e s for l i t t e r s w i t h no s u r v i v i n g pups. * Significantly different from the Group 1 value (p<0.05). ** S i g n i f i c a n t l y d i f f e r e n t from the G r o u p 1 v a l u e (p<0.01). 418-018:PAGE B-67 0005; x PROTOCOL 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 18 VEHICLE CONTROL 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 POOLED PUP WEIGHTS/LITTER (GRAMS) DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+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 [ 11] a 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 [ 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** 6 3 . 5 + 37.4** 68.3 + 38.8** [ 18] a [ 13] a [ 5] a +42.5 + 12.4 +36.3 + 12.7 +28.5 + 17.5 +25.9 + 11.8*1+7.9 + 26.5** -3.2 + 33.0** -9.1 + 35.7* [ 18] a [ 13] a [ 5] a AVERAGE PUP WEIGHT (GRAMS) DAY 1 MEAN+S.D . 6.4 + 0.4 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 + 0.9 DAY 5 MEAN+S.D. 9.8 + 1.1 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN+S.D . +3.4 + 0.9 5.9 + 0.6* 5.9 + 0.6* 5.8 + 0.8** 5.7 + 0.6** 6.4 + 0.5* 6.4 0.8** 6.1 + 0.8** 5.8 + 0.7** 7.0 + 0.7** 6.9 + 1.0** 6.7 + 0.9** 6.2 + 1.0** 7.8 + 0.9* 7.8 + 1.2* t 18] a 7.5 + 1.1** 6.8 + 1.1** [ 18] b t ll] a 8.6 + 1.0** 8.5 + 1.4** 8.1 + 1.3** 7.4 1.4* ( 18] a +2.7 + 0.7* +2.5 + 0.9* +2.2 + 0.8** +1.7 + 1.2* ( 18] a 5.4 + 0.5** 5.4 + 0.5** [ 14] a 5.8 + 0.8** [ 14] a 6.3 + 1.0** [ 14] a 7.1 + 1.1** 1 13] a +1.7 + 1.0** [ 13] a 5.3 + 0.5** t 18] a 5.2 + 0.8** [ 11] a 5.9 + 1.1** [ 7] a 6.5 + 1.4** [ 6] a 7.4 + 1.5** ( 5] a +1.5 + 1.2** t 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). 418-018:PAGE B-68 000572 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 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 MEAN+S.D. 237 13.2 + 2.2 233 1 2 . 9 + 2.3 LIVEBORN MEAN+S.D. N (%) 13.0 + 2.2 234( 98.7) 12.6 + 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 PRES UMED CANNIBALIZED DAY 1 DAYS 2- 5 N / N (%) N / N (%) 1/234 1 0.4) 2/2331 0.8) 22/227( 9.7) 111/205 ( 54.1) VIABILITY INDEX a % N/N 98.7 231/234 41.4 94/227+ DAY(S) = DAY(S) POSTPARTUM a. Number of live pups on day 5 postpartum/nuraber 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 14.1 + 1.4 268( 99. 2) 0.1 + 0.3 2 ( 0 .7) 0 93/268( 34.7)** 172/175( 98.3)** 1. 1 3/268** 418-018:PAGE B-69 c Cfl u i PROTOCOL 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 MEAN+S.D . DAY 2 MEAN+S.D . DAY 3 MEAN+S.D . DAY 4 MEAN+S.D . DAY 5 MEAN+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 MEAN+S.D. 56.2 12.4 49.9 + 10.8 DAY 2 DAY 3 DAY 4 DAY 5 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 56.4 + 12.2 55.8 + 12.3 55.8 + 12.3 55.8 + 12.3 55.0 + 18.0 [ 15) C 49.8 + 12.4 t 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). . o 48.0 + 15.6 54.6 + 41.1 [ 91c 23.3 + 32.5** I 5] c 55.6 + 50.9 I 3]C 50.0 + 70.7 { 2JC 418-018:PAGE B-70 * PROTOCOL 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 MEAN+S.D. 90.6 + 11.6 DAY 3 MEAN+S.D. 99.7 + 14.4 DAY 4 MEAN+S.D. 112.1 + 15.5 DAY 5 MEAN+S.D. 123.9 + 17.1 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN+S.D. +40.9 + 8.2 AVERAGE PUP WEIGHT (GRAMS) DAY 1 MEAN+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 S MEAN+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** l 12] a 63.8 + 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** i 12] a 7.7 + 1.8** ( 12] a +2.0 + 1.4** [ 12] a 4 2 + MEVALONIC ACID 19 57.1 + 22.2** ( lG]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 + 3 3 . 5 [ 2] a 5.2 + 0.3** [ 16] a 6.8 + 5.4** [ 81a,b 5.1 + 1.1** ( 5] a 5 . 8 + 2.2** i 3] a 6.7 + 2.8** [ 2] a +1.3 + 2.6* l 2] a 418-018:PAGE B-71 0Q057S PROTOCOL 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 6 1.6 + CHOLESTEROL DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 17 20 PUPS DELIVERED (TOTAL) N MEAN+S.D . 247 1 4 . 5 + 2.2 271 13.6 + 2.1 LIVEBORN MEAN+S.D. N (%) 14.4 + 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 ) = 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 5 value (p<0.05). ** Significantly different from the Group 5 value (p<0.01). 7 2 + CHOLESTEROL 18 261 14.5 + 1.7 1 4 . 4 + 1.8 259( 99.2) 0.1 + 0.3 2 ( .8) 1 27/258 ( 10.5)** 194/231( 84.0)** 14.3 37/258** 418-018:PAGE B-72 o a PROTOCOL 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 MEAN+S.D . DAY 2 MEAN+S.D . DAY 3 MEAN+S.D . DAY 4 MEAN+S.D. DAY 5 MEAN+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 1 3 . 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. MEAN+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 t 19] C 41.1 + 26.8 [ 14] C 44.4 + 25.0 [ 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 t 7] C 48.6 + 33.8 t 7] C 48.0 + 21.4 t 5] c 418-018:PAGE B-73 000577 PROTOCOL 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 MEAN+S.D. 90.9 + 12.5 DAY 2 MEAN+S.D. 100.7 + 1 8 . 9 DAY 3 MEAN+S .D . 108.7 + 16.8 DAY 4 MEAN+S.D. 123.4 + 17.7 DAY 5 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN+S.D. MEAN+S.D. 134.4 + 20.3 [ 16] b +44.4 + 11.8 [ 16] b AVERAGE PUP WEIGHT (GRAMS) DAY 1 MEAN+S.D . 6.3 + 0.5 DAY 2 MEAN+S.D. 7.1 + 0.8 DAY 3 MEAN+S.D . 7.7 + 0.8 D&Y 4 /A DAY 5 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN+S.D . MEAN+S.D. MEAN+S.D. 8.8 + 0.9 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** [ 14] a 62.5 + 32.4** [ 13] a 73.1 + 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 3 2 . 6 + 27.1** [ 7] a 33.9 + 29.3** [ 7] a 47.9 + 26.2** [ 5] a -24.3 + 24.1** [ 5] a 5 . 3 + 0.4** 4 . 9 + 0.5** I 14] a 5.6 + 0.6** l 7]a 5.8 + 0.8** [ 7] a 6.6 + 0.6** [ 5] a +1.0 + 0.2** t 5] a 418-018-.PAGEB-74 000S78 PROTOCOL 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 NOT NURSING N/N i/i 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 i/i i/i 9 10 11 12 0.8 1 1.2 1.6 17 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. oo 3 418-018:PAGE B-75 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 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 3 1 . 6 + MEVALONIC ACID 18 4 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 oc o PROTOCOL 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 TSSOOO PROTOCOL 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 1 1.2 12 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 1 12 8 4 8 13 17 43 91 2 214 3 6 11 16 39 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( 0.0) PUP INCIDENCE N(%) 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 o oo Cfl 00 (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 22 (PAGE 2): NECROPSY OBSERVATIONS - SUMMARY - FI GENERATION PUPS DOSAGE GROUP DOSAGE MG/KG PROS 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 418-018.`PAGE B-79 000583 PROTOCOL 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 S 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)** 32 ( 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 (%) 17(100.0) 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 (p<0.01). Complete necropsies were not performed on pups in which 418-018:PAGE B-80 000584 PROTOCOL 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 0 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 ( i- 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. SSS'OOO 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-6295.25) TABLE 23 (PAGE 2) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 VEHICLE CONTROL RAT If 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-018:PAGE B-82 c/l 00 a PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR S STUDY NUMBER: T-6295.25) T A B L E 23 (PAGE 3) C L I N I C A L O B S E R V A T I O N S - I N D I V I D U A L D A T A - Fo G E N E R A T I O N F E M A L E R A T S DOSAGE GROUP 2 MEVALONIC ACID CONTROL RAT # DESCRIPTION P o o (ft QO -J 10915 10916 10917 10918 10919 10920 10921 10922 10923 10924 10925 10926 10927 10928 DG ( 1) D G ( 4- 9) D G ( 13- 21) DG( 10- 12) D G ( 17 ) DS( 10 ) D S ( 34 ) D S ( 41- 45) D G ( 0- 2) D G ( 4- 5) DG ( 10 ) D G ( 13 ) D G ( 15 ) D G ( 17 ) D G ( 17 ) D G ( 19 ) D G ( 17 ) D G ( 19 ) D G ( 15 ) D G ( 18 ) DS( 37- 40) DS( 43- 44) DG ( 0) DS( 45 ) DG ( 5) D G ( 18 ) DS( DG ( DG ( DG ( DG ( DG ( DG ( DG( 30 ) 11 ) il ) 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, M O D E R A T E PERIORAL SUBSTANCE EXCESS SALIVATION CHROMORHINORRHEA RED PERIVAGINAL SUBSTANCE RED, SLIGHT PERI O R A L 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. O b s e r v a t i o n c o n f i r m e d at necropsy. DL DAY OF LACTATION 418-018:PAGE B-83 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) T A B L E 23 (PAGE 4): C L I N I C A L O B S E R V A T I O N S - I N D I V I D U A L D A T A - Fo G E N E R A T I O N F E M A L E RATS DOSAGE GROUP 2 MEVALONIC ACID CONTROL RAT 0 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) D G ( 14- 17) D G ( 15 ) DG ( DG ( DG ( DG ( 17 ) 11 ) 18 ) 18 ) D G ( 12- 21) D L ( 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. O b s e r v a t i o n c o n f i r m e d at necropsy. DL = DAY OF LACTATION 418-018:PAGE B-84 oc o Cft 00 00 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) T A B L E 23 (PAGE 5) C L I N I C A L O B S E R V A T I O N S - I N D I V I D U A L D A T A - Fo G E N E R A T I O N FEMALE RATS DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID R A T II DESCRIPTION 10929 10930 10931 10932 10933 10934 10935 10936 10937 10938 10939 10940 . 10941 10942 11529 11530 11531 11532 11533 DS( 40- 42) DS( 40- 44) D G ( 0- 3) DG( 7 ) DG ( 7) DG ( 9) D G ( 13 ) D G ( 15 ) DG ( DG ( DS( DG ( DS( 9- 20) 17 ) 1) 17 ) 2) D G ( 13 ) D G ( 19 ) DG ( 19- 21) DL ( 4) D G ( 13- 21) D G ( 18- 21) D L ( 1- 5) D L ( 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: S C A B (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. O b s e r v a t i o n c o n f i r m e d at necropsy. DL = DAY OF LACTATION Gssoob 418-018:PAGE B-85 PROTOCOL 418-018: OWE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-G295.25) T A B L E 23 (PAGE G ) : C L I N I C A L O B S E R V A T I O N S - I N D I V I D U A L D A T A - Fo G E N E R A T I O N F E M A L E RATS DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID R A T ft DESCRIPTION 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 DL ( 2) D S ( 28- 32) D S ( 14- 18) D S ( 19- 23) DG ( 12 ) DS ( 4) DS ( 4) D S ( 4- 6) DS ( 8- 12) D S ( 12 ) D S ( 12 ) D S ( 12 ) DL ( 3) DL ( 2) D G ( 13 ) D G ( 1G ) DS ( 41- 44) DG ( 0) D G ( 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 ooo C/T CD O 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 23 (PAGE 7) CLINICAL OBS!ERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 4 2 MG/KG PFOS t MEVALONIC ACID RAT It DESCRIPTION Cj J r '*'' C/1 u 10943 10944 10945 10946 10947 10948 10949 10950 10951 10952 10953 10954 DL( 3 ) DL( 5 ) DS( 32 1 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) 1X3( 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 CHROMORH INORRHEA 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 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 V co Jpm5 to 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 1 DS( 18 ) DG ( DG ( 1) 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 PROTOCOL 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 It DESCRIPTION 11556 11557 DL( DG ( DG ( DL( 1) 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 o JA CJ 418-018 :PAGE B-89 PROTOCOL 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 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 11550 11559 c 11560 11561 oo 11562 4P 1 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 ( - 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-018:PAGE B-90 PROTOCOL 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 It DESCRIPTION 11568 11569 11570 11571 DS ( 25 ) DS ( 25- 40) DS ( 42- 44) DG ( 0- 21) DL ( 1- 4) DL ( 2 ) 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 c o o \ in 418-018:PAGE B-' 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 12): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL RAT II DESCRIPTION 10971 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- 45P'"' DG ( 0- 20) " DG ( 4- 20) DG( 7- 20) DL ( l- 2) DL ( 1- 5) DL ( l- 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 DL = DAY OF LACTATION a . Observation conffirmed at necropsy. 96S000 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 ft 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 418-018:PAGE B-93 PROTOCOL 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 14) CLINICAL OD SERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 2 MG/KG PFOS + CHOLESTEROL RAT H DESCRIPTION 10985 10986 10987 10988 10989 10990 10991 10992 10993 10994 10995 10996 10997 10998 11586 11587 11588 11589 11590 11591 11592 1*593 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 88S000 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 tt DESCRIPTION 11595 DL( 1 ) SACRIFICED DUE TO NO SURVIVING PUPS 11596 DG ( 19- 20) LOCALIZED ALOPECIA: BACK DL( 1- 3) LOCALIZED ALOPECIA: BACK a DL( 3 ) SACRIFICED DUE TO NO SURVIVING PUPS 11597 DG ( 1- 22) LOCALIZED ALOPECIA: UNDERSIDE DG( 13- 22) LOCALIZED ALOPECIA: LIMBS DL ( 1- 4) LOCALIZED ALOPECIA: LIMBS DL( 1- 4) LOCALIZED ALOPECIA: UNDERSIDE 11598 DS ( 37- 38) INCISORS: MISSING/BROKEN DL( 3 ) SACRIFICED DUE TO NO SURVIVING PUPS .V 11599 DL( 2 ) SACRIFICED DUE TO NO SURVIVING PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION .a. O b s e r v a t i o n c o n f i r m e d at necropsy. 418-018:PAGE B-95 oo o CA PROTOCOL 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 U 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) MO 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-018:PAGE B-96 009000 PROTOCOL 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 N 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-018:PAGE B-97 000601 PROTOCOL 418-010: 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 ft 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( i- 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 PUPS SWOLLEN SNOUT INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS 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 000602 PROTOCOL 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 19): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 11 1.2 MG/KG PFOS RAT ft DESCRIPTION 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 DS 1 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-018:PAGE B-99 000603 PROTOCOL 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 ) DGI 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 DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-018:PAGE B-100 000604 PROTOCOL 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 ft 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) DSl 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 DL * DAY OF LACTATION a. Observation confirmed at necropsy. S09000 418-018:PAGE B-101 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 24 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 VEHICLE CONTROL RAT NUMBER 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 DAY OF PREGNANCY NECROPSY STATUS DG 21 DG 21 DG 21 DG 21 DG 21 DL 5 DG 21 DG 21 DL S 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 OBSERVATIONS a 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-102 909000 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 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. Cosi 10926 DG 12 P 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 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. 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 CM). 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. b. Appeared normal for their developmental ages. 418-018:PAGE B-103 PROTOCOL 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 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 c o 8- PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 24 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 3 1.6 1 MEVALONIC ACID 10929 10930 10931 10932 10933 10934 10935 10936 DL 5 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DS 2 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. 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 DL 5 p ALL TISSUES APPEARED NORMAL. 11529 DL 5 P ALL TISSUES APPEARED NORMAL. A 11530 DL 5 p ALL TISSUES APPEARED NORMAL. 11531 DG 25 NP ALL TISSUES APPEARED NORMAL. o <7> 11532 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. O 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 PROTOCOL 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. 418-018:PAGE B-106 co . 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 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. c 10955 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. o 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. r 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 PROTOCOL 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 o a (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 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 o 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 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 DS 16 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. 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 PROTOCOL 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. cc 10984 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. o 11572 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS o ALL TISSUES APPEARED NORMAL. p 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. 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-629S.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. 11582 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 o h* C/1 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 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 10986 DG 21 DL 3 P P ALL TISSUES APPEARED NORMAL. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 10987 10988 10989 10990 10991 10992 10993 10994 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DL 3 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. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10995 10996 DL 5 DL 2 P P ALL TISSUES APPEARED NORMAL. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS, STOMACH: CONTAINED PUP TISSUE. ALL OTHER TISSUES APPEARED NORMAL. 10997 10998 11586 DL 5 DG 25 DL 3 P NP p ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 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. 900 418-018 :PAGE B-112 PROTOCOL 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 o m p PROTOCOL 418-018: OHE 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 S 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 o o a 00 PROTOCOL 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 PROTOCOL 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 o o to PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) 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 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 DL 5 DS 81 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DG 25 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 DL 5 P NP P P P P P P P P P NP 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-117 CtoV PROTOCOL 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 1.6 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 11048 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 DL 5 P ALL TISSUES APPEARED NORMAL. 11050 DG 21 P ALL TISSUES APPEARED NORMAL. 11051 DG 21 P ALL TISSUES APPEARED NORMAL. 11052 DG 21 P ALL TISSUES APPEARED NORMAL. c 11640 DL 5 P ALL TISSUES APPEARED NORMAL. O 11641 DL 5 P ALL TISSUES APPEARED NORMAL. Q 11642 DL 5 P ALL TISSUES APPEARED NORMAL. 11643 DL 5 P ALL TISSUES APPEARED NORMAL. 11644 DL 5 P ALL TISSUES APPEARED NORMAL. (0 N 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 PROTOCOL 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 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 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 2 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. OO 11064 DL 3 p SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. o 11065 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 1* 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 PROTOCOL 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 i 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 5 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-121 oco o CA PROTOCOL 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 1) : 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 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) . AB 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-122 000626 PROTOCOL 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 BS. 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) . AB S . WT. = ORGAN WEIGHT. REL. t 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. 418-018:PAGE B-123 0006257 PROTOCOL 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 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 10938 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 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. 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 000628 PROTOCOL 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 AB S . REL. W T . % TBW DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943a P -- - 14 .51 10944 P 282 . D 11.92 10945 P 415. C 16.51 10946 P 421. C 17.22 10947 P 385. C 14 .26 10948a P -- - 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 -- - 16.30 10956a P -- - 16.88 11544a P -- - 13.74 11545a P 11546a P -- -- - 18.81 - 14.56 11547 P 299. D 14.89 11548 NP 11549a P 11550a P 11551a P -- -- -- - 17.01 - 13.74 - 15.29 11552 P 11553a P 11554a P 11555a P 310. -- -- -- D 14.55 - 16.80 - 13 .22 - 13.15 11556a P --11557a P -- - 13.73 - 16.09 -- 4.23 3.98 4.09 3.70 -- 4.54 3.92 4.06 -- 4.05 3.94 -- -- -- -- -- 4.98 -- -- -- 4.69 -- -- -- -- -- 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-125 000629 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 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 AB S . REL. W T . % TBW DOSAGE GROUP 5 CHOLESTEROL CONTROL 10951 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 14.48 *- o 00 00 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 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. 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 000630 PROTOCOL 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 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 AB S . 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) . 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. 000631 31-9 3 0 V d :8lO-8lfr PROTOCOL 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 000632 PROTOCOL 410 018; 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. WT. % 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 . 33 3 . 322 . 325. 334 . 340 . 325. 344 . 303 . 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) . AB 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 000633 PROTOCOL 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 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 ABS. 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 418-018:PAGE B-130 000634 PROTOCOL 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. REL. W T . % 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 ALL WEIGHTS WERE RECORDED IN GRAMS (G) . ABS. 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 S 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. ooo o CJ Cfl 418-018:PAGE B-131 PROTOCOL 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 II) : 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 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) . 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 oo 418-018:PAGE B-132 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 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 AB S . REL. W T . % 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 11046a 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 analyses. 418-018-.PAGE B-133 000637 PROTOCOL 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 1166"6 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) . AB 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-134 000638 PROTOCOL 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 IS 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 11509 11510 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. 282. 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-135 000G39 PROTOCOL 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 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 . 24 3 . 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. 63 70 11 81 418-018:PAGE B-136 O O C 40 PROTOCOL 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. 418-018:PAGE B-137 00C41 PROTOCOL 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 4): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 IS 22 29 36 42a 49 56b 63 70 77 81 RAT It 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 DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 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 . 220 . 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 . 287. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. La3t value recorded before cohabitation. b. First value recorded after cohabitation. 418-018:PAGE B-138 00G42 PROTOCOL 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 U 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 190 . 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 . 266 . 273 . 205. 294 . 306 . 250 . 263. 263 . 271. 237. 258. 257. 265 . 226 . 236. 254 . 250 . 266 . 290. 291. 308 . 256 . 263 . 283 . 289 . 268 . 270. 288 . 296 . 241. 252 . 255 . 255 . 270 . 284 . 296 . 286 . 249. 263 . 280 . 276. 308. 238 . 249. 263 . 256 . 245 . 260. 267 . 256 . 269. 296. 290. 295. 244 . 247. 251. 259 . 265 . 272 . 284 . 280 . 267 . 278 . 290 . 292 . 277 . 292 . 285 . 294 . MORIBUND SACRIFICED ON DAY 16 OF STUDY 274 . 286. 284 . 292 . 247 . 258 . 257 . 270 . 240 . 244 . 244 . 252 . 241 . 248 . 260. 253 . 262. 254 . 271. 273 . 234 . 236 . 244 . 247 . 280 . 290. 296. 310 . 238. 247. 253. 253 . 263 . 270. 286 . 289. 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 Q0Q643 PROTOCOL 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 8 15 22 29 36 42a 49 56b 63 20 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 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 000644 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 7): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 IS 22 29 36 42a 49 56b 63 70 77 81 RAT It 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. Laat value recorded before cohabitation. b. Firot value recorded after cohabitation. 418-018:PAGE B-141 000645 PROTOCOL 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 H 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 . 249. 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 oco & o PROTOCOL 410-018: 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 9): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 RAT It 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 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 S T U D Y a. Last value recorded before cohabitation. b. Firot value recorded after cohabitation. 56b 322. 63 326. 70 316. 77 317. 81 322. 418-018:PAGE B-143 000647 PROTOCOL. 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.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 tt DOSAGE GROUP 10 1 MG/KG PFOS 11019 11020 11021 11022 11023 11024 11025 11026 11027 11020 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 . 256. 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 000G48 PROTOCOL 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 RAT It DOSAGE GROUP 11 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 203 . 228 . 227 230 248 214 . 251 . 264 275 272 206 . 233 . 247 257 273 216. 231. 250 260 265 206 . 223 . 235 246 254 212 . 231. 241 261 268 209. 223 . 233 237 249 213 . 249. 255 267 267 195. 214 . 219 234 236 212 . 238 . 247 254 258 202 . 210 . 224 232 231 237. 245 . 263 267 271 221. 222 . 239 248 252 223 . 243 . 255 272 275 210. 228 . 243 251 269 239. 254 . 257 257 273 219. 231 . 231 241 249 223 . 236 . 239 248 250 208 . 219. 217 228 231 228 . 253 . 257 258 262 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 36 42a 49 1.2 MG/KG PFOS 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. 56b 63 70 322 . 318 . 306. 77 305. 81 308 . 418-018:PAGE B-145 000C49 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 12): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 8 15 22 29 36 42a 49 56b 63 RAT It DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11040 11041 11042 11043 11044 11045 11046 11047 11048 21049 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. 288 . 281. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 70 277. 77 81 275 . 272 . 418-018:PAGE B-146 000C50 PROTOCOL 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 15 22 29 36 42a 49 56b 63 70 77 81 RAT K DOSAGE GROUP 13 2 MG/KG PFOS 11053 11054 11055 11056 11057 11058 11059 11060 11061 11062 11063 11064 11065 11066 1X654 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 ooorsi, PROTOCOL 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 . 2 94 . 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. 289. 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. 418-018:PAGE B-148 000652 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 2): 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 RAT 0 10901 P 10902 P 10903 P 10904 P 10905 P 10906 P 10907 P 10900 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 DOSAGE GROUP 1 312 . 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. 202 . 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. VEHICLE CONTROL 352. 337. 380. 385. 362. 394 . 379. 363 . 349. 388 . 374 . 340. 370. 312. 320. 411. 366 . 305 . 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 . 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. 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 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-149 0Q06S3 PROTOCOL 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 It 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 1092B 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 93000 PROTOCOL 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 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 ft 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. 402. 388. 411. 426. 452. 346. 358. 366. 383. 386 . 393 . 420. 443 . 345 . 362 . 375. 391. 379. 394 . 412 . 426. 372 . 388 . 398. 422. 356. 370. 385. 393. 340 . 349 . 362 . 385. 359. 375. 381. 400. 376 . 390. 405 . 428. DEATH WAS ATTRIBUTED TO AN INTUBATION1 ACCIDENT 398. 412. 433 . 351. 367. 386. 408. 304 . 317. 332. 340. 390. 407. 427. 436. 368. 374 . 386. 393. 380. 404 . 413 . 422. 371. 387 . 405 . 406. 358 . 366 . 372. 386. 361. 375. 378. 387. 306 . 298. 295. 302. 304. 304. 340. 355. 376. 382. 339. 346. 355. 369. 388. 402. 418. 421. 358. 433. 384. 403. 363. 370. 384 . 393. 377. 390. 408. 416. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 24 25 302 . 307. 418-018 :PAGE B-151 000C55 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 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 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 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 9 266 . 289. 267. 282 . 252 . 296. 255 . 284 . 307. 249. 308 . 260. 291. 265. 328. 256 . 294 . 272 . 314. 290. 290. 275. 281. 241. 320. 272 . 299. 328 . 10 274 . 295. 274 . 283 . 250. 293 . 258. 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. sfcooo 418-018:PAGE B-152 PROTOCOL 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 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 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 25 256 . 256. 267 . 267. 418-018:PAGE B-153 000C57 PROTOCOL 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 tt 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. 2B4 . 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 . 418-018:PAGE B-154 8S2000 PROTOCOL 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 tt 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 . 2B0 . 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. 2S4 . 373 . 330. 388 . 359. 374. 353. 385. 361. 388 . 360. 354. 396. 385. 349 . 405 . 276 . 380 . 3 76. 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 . 418-018:PAGE B-155 000059 PROTOCOL 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 9): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 RAT If 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. 292 . 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 000G60 PROTOCOL 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 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. 394 . 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 . 3 92. 394 . 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 21 378. 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. 22 436. 314 . 405. 296. 23 314 . 294 . 24 312 . 293 . 25 318. 302. 418-018:PAGE B-157 000G61 PROTOCOL 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 i RAT tt DOSAGE GROUP 6 10971 P 10972 P 10973 P 10974 P 1097S P 10976 P 10977 P 10970 P 10979 P 10980 P 10981 P 10982 P 10903 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 . 2 236 . 293 . 249. 280 . 269. 200. 256 . 284 . 282 . 265 . 251. 259. 272 . 287. 223 . 287. 266. 269. 272. 297 . 262 . 314 . 282 . 284 . 275. 268 . 282. 259. 3 242 . 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 . 4 245. 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 . 56 78 1.6 MG/KG PFOS + CHOLESTEROL 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 o ooo & to 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 . 2 93 . 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. 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-629S.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 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 22 23 418-018:PAGE B-159 U00C63 PROTOCOL 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 II 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 DOSAGE GROUP 7 257. 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 . 258257. 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. 2 MG/KG PFOS + CHOLESTEROL 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. 2 90. 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. *99000 418-018:PAGE B-160 PROTOCOL 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 0 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. 3 66. 368. 308 . 378. 347. 347. 394 . 382. 379. 354 . 346. 266. 361. 423 . 359. 3 97. 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 25 274 . 283. 418-018:PAGE B-161 000C65 PROTOCOL 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 3456 RAT ft 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 10 11 12 291. 334 . 296. 304 . 312. 313 . 328. 313. 343 . 317 . 320. 344. 279. 318. 295. 344 . 300. 289. 326. 331. 297. 331. 298 . 314 . 307. 324. 327. 319. 348. 328. 328. 350. 281. 322. 301. 341. 298 . 292. 330. 338. 303 . 338 . 303 . 307 . 312 . 337. 330. 321. 352. 332. 334 . 355. 285 . 329. 306. 346. 309. 303 . 342 . 339. 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 U00C6G PROTOCOL 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 RAT H 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 , 3 69, 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 . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY DAY OF PRESUMED GESTATION 20 379. 422 . 392. 403 . 416. 422 . 397. 417 . 444 . 403. 423. 449. 352. 430 . 386. 442 . 380. 386. 419. 444 . 21 389 428 401 414 434 412 430 459 423 438 469 350 399 457 395 409 429 446 418-018:PAGE B-163 c oo 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 17) : MATERNAL BODY HEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT tt DOSAGE GROUP 9 0.8 MG/KG PFOS 11009 P 11010 P 11011 P 11012 P 11013 P 11014 NP 1101S 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 . 250 . 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 000G6S PROTOCOL 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 PREGNANCY STATUS DAY 13 14 15 16 17 18 19 RAT 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 DOSAGE GROUP 9 274 . 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 . 0.8 MG/KG PFOS 328. 403 , 398 344 , 337 330. 417 . 426 . 366. 349. 371 339. 302 , 368. 300, 416. 354 , 403 . 388. 383 359. 379. 385. 356 388 . 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. 22 289. 339. 306 . 23 296. 312 . 24 294 . 308 . 25 298. 297. 690000 418-018-.PAGE B-165 PROTOCOL 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 i 2 3 4 5 6 RAT ft 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. 272 . 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 c> o PROTOCOL 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 It 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 21 398. 419. 402. 303 . 386. 404 . 381. 402. 377. 436. 369. 421. 396. 393 . 404 . 444 . 377. 381. 414 . 400. 433 . 412. 304 . 420. 399. 411. 381. 402. 412. 454 . 395. 370. 414 . 22 310. 403 . 23 307. 24 311. 25 309. 418-018:PAGE B-167 c 8 C> PROTOCOL 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 21): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT (1 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 . 250. 268 . 270 . 237 . 243 . 245. 272 . 277. 282 . 253 . 302 . 286 . 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 . 3IB . 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 00S72 PROTOCOL, 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 H 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 . 394 . 401. 366. 446 . 374. 408. 327. 287 . 361. 401. 431. 433. 394 . 363. 22 290. 23 24 288 . 291. 2S 300. 418-018:PAGE B-169 0Q0673 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 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 000G74 PROTOCOL 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 tf 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 11040 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. 4X0. 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 . 374 . 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. 418-018:PAGE B-171 000C75 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 25): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 RAT tt DOSAGEIGROUP 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 000G76 PROTOCOL 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 tt 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 24 25 264 . 266 . 274 . .eboo 418-018:PAGE B-173 PROTOCOL 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 H 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 c o o 00 PROTOCOL 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 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 . 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-175 000G79 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 28 (PAGE 3): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT H 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 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 Q00C80 PROTOCOL 4 1 8 - 0 1 8 : 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 N DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 10944 10940 10952 10955 10956 11544 11545 11546 11547 11548 11549 11550 11551 115S2 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 PUPS 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 ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-177 QOOCbX PROTOCOL 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 11560 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 oc on N 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 28 (PAGE 6): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT It DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 10973 10977 10979 10983 10904 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 o o c ocr U 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 7): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT It 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 LACTATION DUE TO NO SURVIVING PUPS 317 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 277 . 282. 291 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 250 . 261. NOT PREGNANT 279 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 340 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 312 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 303 . 290. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 290. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 336. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 312. 297. 272 . 262 . 309. 299. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 291. SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 282 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 278. 275 . 287 . SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 278 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 418-018:PAGE B-180 c o & 05 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 28 (PAGE 8) : MATERNAL BODY WEIGHTS - LACTATION INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 rat # DOSAGE GROUP B 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-181 oo a oo Cfl PROTOCOL 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): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT K 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 418-018:PAGE B-182 Ci 00 ' a PROTOCOL. 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) T A B L E 28 (PAGE 10): M A T E R N A L B O D Y W E I G H T S - L A C T A T I O N - I N D I V I D U A L D A T A - Fo G E N E R A T I O N FEMALE R ATS DAY 1 5 R A T tt DOSAGE G R O U P 10 1 MG/KG PFOS 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 11627 n e 28 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. A L L W E I G H T S W E R E R E C O R D E D IN G R A M S (G) . DAY = DAY OF LACTATION 418-018:PAGE B-183 oo o o 00 PROTOCOL 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 HEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 RAT K DOSAGE GROUP 1 11029 11030 ]1031 11032 11033 11034 11035 11036 1103V 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 oo o 00 00 PROTOCOL 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 11019 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 o c car l'ROTOCOL, 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 13) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT It 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 2 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 1 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 o n <x> PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'1S 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 (I 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. 148 . 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 . 111. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF STUDY a. La3t value recorded before cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. 418-018:PAGE B-187 000691 PROTOCOL 418-018: OWE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-62 95. 25) TABLE 29 (PAGE 2) : FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 418-018:PAGE B-188 CCTO\ DAYS 1- 8 8- 15 15- 22 22 - 29 29- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT H 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 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 . 99. 106 . 97. 112 . 107 . 119. 119. 94 . 104 . 122 . in. 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. 269000 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-G295.2S) TABLE 29 (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 U DOSAGE: GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID }0929 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 . i n . 132. 135 . 138 . C 133 . 121. 123 . i n . 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 C69000 PROTOCOL 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 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 ft 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. ALL WEIGHTS WERE RECORDED IN GRAMS (G) . DAYS = DAYS OF STUDY a. Last value recorded beEore cohabitation. b. First value recorded after cohabitation. c. Spilled feed precluded the calculation of this value. d. Value was not recorded. 418-018:PAGE B-190 000694 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 29 (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 II DOSAGE GROUP 5 CHOLESTEROL CONTROL 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 1155B 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 thi value. 418-018:PAGE B-191 000G9S PROTOCOL. 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 tt 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 000S96 PROTOCOL 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 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 . 111. 96 . 113 . 108 . 114 . 107. 96. 106. 111. 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. 418-018:PAGE B-193 000G97 PROTOCOL 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): 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 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 . 139. 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 . 1i7 . 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. 869000 418-018:PAGE B-194 PROTOCOL 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 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 It 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. 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-195 c o Ci PROTOCOL 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 It 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 thi3 value. 418-018 :PAGE B-196 oo o6 PROTOCOL 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 tt 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 . Ill . 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 Q00701 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 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 . 143 . 120. 135 . 132 . 101. 106. 94. 97. 59. 130. 131. 137 . 137. 120. 101. 144 . 134 . 140. 132 . 132 . 101. 143 . 137. 141 . 130. 127. 100. 126. 126. 127. 131. 135. 94 . 138 . 136. 139 . 125 . 128 . 105 . 147. 131. 139. 143 . 119. 100. 136. 130 . c c 131. 93 . 122 . 116 . 116 . 129. 116. 97. 116 . 115 . 120 . 125 . 117 . 91. 125 . 127 . 138 . 132 . 126. 101. 143 . 130. 128 . 134 . 116. 95. 133 . 122 . 122. 120. 109. 99. 137 . 131. 138 . 135 . 121. 93 . 155. 127. 124 . 133 . 127. 102 . 156 . 160. 163 . 146. 148. 116. 142 . 138 . 153 . 128. 121. 100 . 148 . 136 . 140 . 123 . 132 . 97 . 128 . 126. 117. 114 . 133 . 91. 173 . 154 . 160. 146 . 179. 128 . 137. 131. 128 . 116 . 118 . 113 . 144 . 141. 135 . 130. 140 . 115. 103 . 98 . 104 . 95 . 108 . 90. 149. 154 . 162 . 147. 133 . 123 . 129. 129. 119. 126. 105. 91. 149. 150. 141 . 133 . 149. 113 . 148. 146. 141. 142 . 151. 108 . 158. 112 . 121. 115 . 116. 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. 20^000 418-018:PAGE B-198 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 29 (PAGE 13): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1- 8 8- 15 35- 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. Ill. 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 in. 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. Spilled feed precluded the calculation of this value. 418-018:PAGE B-199 <300703 PROTOCOL 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 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 H 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 waa not recorded. 418-018:PAGE B-200 000704 PROTOCOL 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 - 3 4 1 4 - 2 1 21 - 25 RAT It 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 154 . 163 . 160 . 185. 132 . 138 . 157 . 173 . 150 . 152. 148 . 184 . 184 . 180. 155 . 155. 159. 166. 134 . 106. 57. 138. 154 . a 162 . 188 . 191. a 153 . 155 . 176 . 166 . 169. 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 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-201 00070S 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 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 # 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 wag not recorded. b. Spilled feed precluded the calculation of this value. 418-018:PAGE B-202 PROTOCOL 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 4) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 1 4 1 4 - 2 1 2 1 - 2 5 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 11540 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 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 000707 PROTOCOL 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 cc ^ o 00 PREGNANCY STATUS DAYS 0 - 7 7 - 1 4 1 4 - 2 1 21 - 25 RAT tt 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 . 153 . 168 . 152 . 159 . 184 . 160 . 165 . 198 . 185 . 210. 215. 174 . 171. 191. 153 . 153 . 172 . 183 . 178 . 207. 161. 183 . 203 . 144 . 142 . 167 . a a 174 . 170 . 165. 153 . 160 . 166 . 152 . 184 . 164 . 193 . 178 . 126 . 142 . 161. 191. 196 . 176. 140 . 127. 149. 69. 191 . 177. 162. MORIBUND SACRIFICED ON DAY 16 OF STUDY a 203 . 192 . 159. 156 . 156. 155 . 156 . 168 . 178 . 153 . 114. 74. 176 . 182 . 207. 135 . 148 . 152 . 161 . 167. 167. 132 . 146 . 146 . 186 . 183 . 203 . 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 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 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 tt 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 000709 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 7) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT t) 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 ooorio PROTOCOL 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 H 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 . 149. 133 . 166 . 161. 170 . 168 . 147. 163 . 155 . 180 . 165. 177 . a 153 . 170 . 169. 148 . 161 . 151. 169 . 153 . 147 . 155. 180. 141. 374 . 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. TT -'.OOO 418-018:PAGE B-207 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSORS 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 000712 PROTOCOL 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 H 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 . 145 . 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 000713 PROTOCOL 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 11) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 1 4 H - 21 21 - 25 RAT It 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 PROTOCOL 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 c e 8 PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT U 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. 73 . 73 . 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-211 PROTOCOL 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 tt 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 . 145 . 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 . 63. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value wag not recorded. b. Spilled feed precluded the calculation of this value. 418-018:PAGE B-212 000716 PROTOCOL 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 H DOSAGE GROUP 1 VEHICLE CONTROL 10906 10909 10911 10913 10914 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 121. 113. 116. a 119. 86 . 104 . 98 . 150. 148 . 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. TiOOO 418-018 :PAGE B-213 PROTOCOL 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 ft 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 8T 1000 418-018:PAGE B-214 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 31 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT (t DOSAGE GROUP 3 L.C M G /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 value. 418-018:PAGE B-215 000713 PROTOCOL 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 4) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT U 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 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 1 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 1 OF LACTATION DUE TO NO SURVIVING PUPS ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION 418-018:PAGE B-216 %* 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 5): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT U DOSAGE GROUP 5 10957 1096G 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 i 139. 99. 129. NOT PREGNANT 124 . 118. 124 . 97. 131. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION CHOLESTEROL CONTROL 418-018:PAGE B-217 0007Z I 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 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 0007322 PROTOCOL 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 It DOSAGE GROUP 7 2 MG/KG PFOS -t CHOLESTEROL 10986 10994 10995 10996 10997 10998 115B6 11587 11588 11589 11590 11591 11592 11593 11594 11595 11596 11597 11598 11599 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 56 . SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 83 . NOT PREGNANT 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 53. SACRIFICED ON DAY 5 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 67. 39. 50. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF 1ACTATION DUE TO NO SURVIVING PUPS 61. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 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-219 000723 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 31 (PAGE 8) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT # DOSAG 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. 145 . 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 000724 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 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-221 000725 PROTOCOL 418-01B: 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 tt 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 0 0 0 /c 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 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-223 000727 PROTOCOL 438-018: 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 12): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 RAT K DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11041 11042 1i04 3 11048 1X049 11640 11641 11642 11643 11644 1164 5 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 821000 418-018 :PAGE B-224 PROTOCOL 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 % PROTOCOL 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 RAT # DAYS IN COHABITATION 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 2 2 3 i 1 3 4 i 3 3 3 1 13 4 2 1 4 5 5 2 1 3 1 1 1 1 1 1 M = MATED C = CONFIRMED P = PREGNANT VEHICLE CONTROL 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 NP P p P P P P P p p p p p p p p p O C '000 418-018:PAGE B-226 PROTOCOL 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 H 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 DAYS IN COHABITATION 1 2 3 4 4 2 3 6 4 4 2 i 12 2 2 3 3 1 1 3 2 3 1 2 i 4 1 1 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 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 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 TVS. ooo 418-018:PAGE B-227 PROTOCOL 4X8-018: OHE 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 FERTIL1TY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID RAT 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 1M FOUND DEAD ON DAY 2 OF STUDY 3M 3M 3M 2M 3M 3M 4M iM iM iM iM 4M iM iM iM 2M iM iM iM 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 ooo V Cj 0 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 PROTOCOL 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 RAT ft DAYS IN COHABITATION 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 1 4 3 3 2 5 3 3 3 12 3 1 3 4 1 1 1 3 4 2 4 1 i 2 2 1 2 4 M = MATED C = CONFIRMED P = PREGNANT 2 MG/KG PFOS + MEVALONIC ACID 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 NP P P P P P P P P P 418-018:PAGE B-229 V C IO O O PROTOCOL 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 S 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 W M OOO PROTOCOL 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 RAT K DAYS IN COHABITATION 10971 10972 10973 10974 10975 10976 10977 10978 10979 10900 10981 10982 10983 10984 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 3 3 1 2 1 3 3 3 3 4 1 1 4 4 1 1 2 2 1 1 2 4 i i 1 1 2 i M = MATED C = CONFIRMED P = PREGNANT 1 .6 MG/KG PFOS + CHOLESTEROL 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 p p 418-018:PAGE B-: K> 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) TABLE 32 (PAGE 7): MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 7 RAT II DAYS IN COHABITATION 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 3 3 3 3 1 4 3 4 2 3 2 3 2 2 1 i 5 2 2 1 13 i i i 1 i 3 4 M MATED C = CONFIRMED P = PREGNANT 2 MG/KG PFOS + CHOLESTEROL 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 NP p p p p p p p p p p p p p p 418-018:PAGE B-232 000736 PROTOCOL 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 RAT It DAYS IN COHABITATION 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 3 3 2 4 3 3 3 4 3 4 3 2 i 2 2 4 1 3 3 1 M MATED C = CONFIRMED P = PREGNANT 0.4 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 P P P P P P P P P P P P P P P P P V'OOO 418-018:PAGE B-233 PROTOCOL 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 RAT # DAYS IN COHABITATION 11009 11010 non 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 3 4 2 3 4 14 4 1 3 5 1 1 1 2 2 4 3 1 4 1 M = MATED C = CONFIRMED P = PREGNANT 0.8 MG/KG PFOS MATING STATUS M M M M M DID NOT MATE 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 PREGNANCY STATUS P P P P P P P NP P NP P P P P P P P P P scrooo 418-018:PAGE B-234 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 32 (PAGE 10) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 10 1 MG/KG PFOS RAT tf DAYS IN COHABITATION MATING STATUS MATING DATE 11019 14 M C 11020 3M c 11021 i M c 11022 6 M c 11023 3 M c 11024 1M c 11025 5 M c 11026 4 M c 11027 iM c 11028 3 M c 11620 2 M c 11621 1M c 11622 1M c 11623 4 M c 11624 2M c 11625 1M c 11626 2 M c 11627 3 M c 11628 1 M c 11629 2 M 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 418-018:PAGE B-235 00079 PROTOCOL 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 RAT It DAYS IN COHABITATION 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 4 14 4 2 2 i 2 3 1 3 2 2 1 2 1 1 1 i 2 1 M = MATED C * CONFIRMED P PREGNANT 1.2 MG/KG PFOS MATING STATUS M DID NOT MATE 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 PREGNANCY STATUS P P P P P P P P P P NP P P p p P p P P o?:oon 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 RAT It DAYS IN COHABITATION 11039 11040 11041 11042 11043 11044 11045 11048 11047 11048 11049 11050 11051 11052 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 14 I 4 4 2 1 1 3 3 4 5 1 3 1 3 3 2 1 2 1 1 1 4 1 13 4 3 1 M = MATED C = CONFIRMED P = PREGNANT 1.6 MG/KG PFOS MATING STATUS 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 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 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 PROTOCOL 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 13) : MATING AND FERTILITY AND DAYS IN COHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 13 RAT tt 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 i 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 00742 PROTOCOL 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 il 4 IS 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 8 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 IS 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 1 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 0 4 04 0 0 0 il 3 14 11 5 16 5 9 14 0 0 0 0 0 0 0 0 0 5 9 14 8 11 19 6 9 15 0 0 0 0 0 0 0 0 0 6 9 15 11 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 0 0 0 1 0 1 0 0 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 11 0 0 0 2 0 2 0 0 0 9 4 13 9 4 13 7 12 19 0 0 0 0 0 0 0 0 0 7 12 19 7 12 19 PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-018:PAGE B-239 000743 PROTOCOL 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 tt 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 ii 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 00 01 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 00 010 i 0 0 0 10 5 15 10 5 15 7 6 13 0 0 0 1 i 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 1 0 0 0 7 8 15 11 8 19 11 5 16 0 0 0 0 0 0 0 0 0 11 5 16 11 S 16 5 8 13 0 0 0 1 0 1 0 0 0 6 8 14 8 12 20 ii 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 0 0 1 0 1 0 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 11 18 7 11 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 17 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 a 9 17 PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-018:PAGE B-240 000744 PROTOCOL 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 FETUSE5 DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA RAT tt 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 1098S 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 11 16 10 4 14 0 0 0 1 1 2 0 0 0 11 5 16 11 5 16 X230000000001231 34 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 11 19 6 3 9 0 0 0 2 1 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 ii 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 ii 15 0 0 0 0 0 0 0 0 0 4 11 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 8 14 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. S fclO O 418-018:PAGE B-241 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 34 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS NUMBER OF LIVE POOLED FETAL FETUSES BODY WEIGHT (G) -- - (CONCEPTUSES DEAD OR RESORBED RAT # TOTAL TOTAL N N DOSAGE GROUP 1 VEHICLE CONTROL 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 14 1 7.1 13 1 7.7 15 1 6.7 15 1 6.7 14 1 7.1 17 1 5.9 15 1 6.7 16 4 25.0 DOSAGE GROUP 2 MEVALONIC ACID CONTROL 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 16 2 12.5 17 1 5.9 14 4 28.6 14 0 0.0 15 0 0.0 IS 6 40.0 16 0 0.0 16 1 6.2 DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10930 13 10931 13 10932 15 10933 16 10934 9 10935 12 1 10937 11 1 10940 19 62.00 65.26 74.42 82.94 40.86 64.44 55.49 86.00 15 2 13.3 14 1 7.1 16 1 6.2 18 2 11.1 17 8 47.0 12 0 0.0 13 2 15.4 19 0 0.0 418-018:PAGE B-242 000746 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 34 (PAGE 2): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS NUMBER OF LIVE FETUSES RAT tt TOTAL DOSAGE GROUP 4 10945 10946 10947 10949 10950 10951 10953 10954 16 15 15 8 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 NN 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 1 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 5.9 418-018:PAGE B-243 0077 PROTOCOL 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 1098S 10987 10988 10989 10990 10991 10992 10993 14 ii 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 Nt 2 MG/KG PFOS + CHOLESTEROL 14 0 0.0 15 4 26.7 16 2 12.5 3 0 0.0 14 1 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 1 6.7 17 3 17.6 11 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 6.2 418-018:PAGE B-244 000748 PROTOCOL 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 tf 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 CLa 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 It CLa 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 t) CL3 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 E A A A A A/ A A A A A A A/ E A A A A E A A E A A/ A E E E A E 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 E = EARLY RESORPTION L = LATE RESORPTION A A AAA A A AA A AAAAA A A A AAA A E EAEAAA AA AAA A AAAAAA '7 II DENOTES POSITION OF CERVIX A AA CLs CORPORA LUTEA/OVARY GfrtOOO 418-018:PAGE B-245 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 35 (PAGE 2) : FETAL VITAL STATUS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS FETUS K 1 DOSAGE GROUP 4 RAT # CL3 10945 11/ 6 10946 12/ 6 10947 11/ 5 10949 11/10 10950 10/ 7 10951 10/13 10953 10/ 5 10954 9/10 A A A E A A E L DOSAGE GROUP 5 RAT # CL9 10958 8/ 7 10959 10/10 10960 11/ 8 10961 11/ 5 10962 8/12 10963 11/ 8 10964 10/10 10965 10/ 3 A A A A A A A A DOSAGE GROUP 6 RAT ft CLs 10972 11/12 10974 7/11 10975 6/ 6 10976 8/ 8 10978 11/ 8 10980 11/11 10931 7/ 7 10982 8/ 9 A A A A A A A A A = ALIVE D = DEAD 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 2 MG/KG PFOS + MEVALONIC ACID 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 A A A A A A A A A A/ A A A A A E A A A A A A A/ E E E E E E A E A A A E A A A A A/ A A A A A E E A E A A A 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 E A A A A A/ A E A A A A A CHOLESTEROL CONTROL 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 A A A/ A A A A A A A E 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 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 A A A A E A A A A A A/ A A A 1.6 MG/KG PFOS + CHOLESTEROL 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 O S ',0 0 0 418-018: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 H 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 ft 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 # CL9 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 It 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 /" D E N O T E S P O S I T I O N O F C E R V I X C L s = C O R P O R A L U T E A / O V A R Y 418-018:PAGE B-247 T S '000 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 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 234 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 B 15 14 14 li 15 10 6 68 89 6 11 37 74 96 69 4 11 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:PAGEB-248 000752 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 1 MF NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 1921 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 11 5 8 9 3 7. 7 3 8 6 4 4 4 6 95 95 95 95 85 75 75 75 11 3 11 3 11 3 11 3 85 85 85 85 WAS ATTRIBUTED1TO AN INTUBATION ACCIDENT 6 11 6 11 6 11 6 11 75 75 75 75 68 68 68 68 69 69 69 69 93 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 000753 PROTOCOL 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 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 MF NUMBER OF LIVE PUPS AT COMPLETION OF' DAY POSTPARTUM 234 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 il 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 i2 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 IMPLAN TATIONS N 13 15 14 15 17 15 14 10 17 14 11 13 14 9 15 13 16 16 418-018:PAGE B-250 OOO S 4 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.2S) 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 234 MF MF MF 5 MF 112 -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-251 000 ;55 PROTOCOL 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 5): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP S 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 234 MF MF MF 5 MF 10957 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11566 11569 11570 11571 23 11 0 il 29 22 15 0 15 78 23 15 0 15 87 22 15 0 15 87 22 16 1 17 88 23 17 0 17 89 23 14 0 14 10 4 23 18 0 18 7 il 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 11 55 23 14 0 14 86 23 16 0 16 88 23 12 0 12 75 23 11 0 11 38 29 78 87 87 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 11 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 88 75 38 29 78 87 87 87 88 10 4 7 il 12 4 87 4 10 59 S5 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-252 000/56 i'ROTOCOL 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 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 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 10971 10973 10977 10979 10983 10984 11572 31573 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 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 34 76 75 a4 12 3 79 64 49 26 84 48 88 66 97 96 66 39 3 il 2 li 65 23 54 41 84 12 3 67 12 31 26 84 48 166 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 13 418-018:PAGE B-253 000 *c/1v 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 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 2 34 MF MF MF 5 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 mi 14 10 15 17 87 78 78 58 12 4 76 76 76 310 5 88 11 4 48 97 -10 4 44 il 4 41 74 144 -11 4 14 1-223 -53 25 -1 -133 11 "* .- -43 -il 4 ---1--23 15 -1 -- -- 32 --- -- -- 33 -11 4 -- --1-- -23 15 -1 --32 -- -- --33 -11 4 ------23 15 ---32 --- M = MALE F = FEMALE ( ) NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. I ) = 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 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 M1 F NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 10999 11000 11001 11002 11003 11004 1100S 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) 1 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 6 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 s i PROTOCOL 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 (DAYSi N LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN 1 M[ F NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 MF MF MF 5 MF 11009 11010 11011 11012 11013 11014 11015 11016 11017 11010 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 11 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 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 IMPLAN TATIONS N 14 20 16 16 16 18 13 12 15 13 15 18 13 15 17 13 13 418-018:PAGE B-256 coo 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 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 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 14 15 17 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 11 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 a7 5 12 67 62 5 12 66 10 6 89 77 5 10 94 84 3 11 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 13 418-018:PAGE B-257 c e $ I* 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 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 M1 F NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 2 34 MF MF 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 54 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 IMPLAN TATIONS N 16 17 16 16 15 14 18 17 19 7 13 16 14 15 16 16 14 16 418-018:PAGE B-258 000762 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 12): HATURAL 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 M1 F NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 234 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 ii 77 77 32 88 58 88 48 85 64 38 47 56 88 67 53 95 6 10 -- . 21 88 46 88 15 74 -- 37 -2 46 31 57 52 85 7 -- . 2i 88 46 88 13 74 -37 -i 46 257 52 84 5 -- 21 78 46 88 12 74 -37 -i 46 257 42 84 5 - __ 21 78 46 88 -2 64 -37 -i 46 -57 42 84 M = MALE F - FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLAN TATIONS N 17 16 15 10 15 15 17 12 14 15 12 13 15 18 13 12 14 418-018:PAGE B-259 3oc o i t) 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 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 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 11055 11057 11059 11061 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 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) 1 14 10 15 16 10 14 14 14 12 13 14 6 14 15 13 16 11 16 IS 58 45 11 4 79 73 68 59 4 10 46 32 -2 -- 95 12 94 86 63 6 10 22 22 -- il 2 22 1- -- 54 48 46 1- -- -- 12 -- 21 --- 23 -- 2- -- il 2 ---- 44 48 46 ------ -1 --- 11 -- -- -- 11 2 ---- 44 47 46 ------ -1 --- 11 -- -- -- il 2 --- -- 44 47 46 --- -- --- -1 ---- -- M = MALE F = FEMALE ( ) = NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. TOTAL IMPLANTATIONS N 15 11 18 16 11 14 14 15 12 16 15 14 15 15 13 16 12 17 15 418-018:PAGE B-260 c oo a PROTOCOL 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 11S04 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 94.8 07.1 108.0 112.3 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 94.9 111.2 107.9 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 96.7 134.0 127.2 117.5 111.1 146.5 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-: o X (A N> 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 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 11520 84.2 92.0 99.6 115.9 122.0 80.3 88.9 92.6 107.3 119.9 89.9 96.5 107.3 119.5 132.7 87.0 91.0 95.4 103.1 108.0 MORIBUND SACRIFICED ON DAY 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 95.7 103.3 116.1 131.5 144.9 74.0 82.2 93.2 107.1 120.6 93.0 102.8 116.6 130.0 146.7 90.3 96.7 108.7 121.8 134.7 79.0 90.1 97.7 106.7 117.3 73.8 80.5 89.5 101.4 110.8 90.1 93.2 101.5 115.1 129.9 47.0 54.4 53.6 63.0 71.8 NOT PREGNANT 94.4 103.5 117.1 131.5 145.3 79.2 88.2 97.8 110.6 119.7 85.8 91.9 103.0 112.3 126.4 80.7 87.3 94.6 107.0 123.6 75.9 84.3 97.5 109.8 124.1 93.6 104.9 112.2 123.9 132.5 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-262 ooc Ch a PROTOCOL 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 PUPS 95.1 104.3 64.5 71.6 77.9 85.4 NO SURVIVING PUPS 37.1 42.4 55.5 63.0 93.4 135.8 31.2 35.2 NO SURVIVING PUPS 9.6 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 oc 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 37 (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-264 768 ooo 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 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 1 1 1 .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 98.3 111.9 147.7 104.3 113.7 106.1 163.6 142.1 120.3 117.5 130.3 137.4 173.5 157.2 152.3 127.0 148.1 107.6 125.0 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 recorded. 418-018:PAGE B-265 000769 PROTOCOL 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 G): 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 PUPS 4.2 NO SURVIVING PUPS 29.7 36.4 39.8 71.8 77.0 83.3 68.6 78.0 85.2 NO SURVIVING PUPS NO SURVIVING PUPS NO SURVIVING PUPS 33 .1 36.5 40.0 91.8 105.1 112.9 81.6 94.0 102.2 NO SURVIVING PUPS 81.8 92.4 99.6 9.2 10.1 10.0 60.2 65.6 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. 000 418-018:PAGE B-266 o PROTOCOL 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 PUPS 81.2 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 38.6 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. 000 418-018:PAGE B-267 *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 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 88 .0 80 .9 59 .3 81..6 77..4 86..1 84 .i 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 .1 91..8 77..5 96,.3 119..5 121,.0 97 ,1 100 .7 131..i 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 ,i 147 .0 74 .1 82 .2 94 ,.6 118..3 80,.7 122 .5 a 117 ,.5 129 .8 118..1 119 .0 124 .6 115 .5 92 .0 118..6 14 6..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 1[G). POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. a. Value was not recorded. 000 418-018:PAGE B-268 'i (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 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 1 1 1 ,5 93 ,9 84 ,6 89..7 73 .5 80 .7 93 .8 84 .7 111 .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. i 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 1(G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-269 000773 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 Group 1 2 3 4 5 a. Dosage Number Administered Identification Date Day o f Study* Early or Late Rat Number 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 -1 4 2 10901 - 10914 10 NOV 00 DG 19 2 10913 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 1.6 mg/kg 16 SEP 00 DS 19 2 10930 PFOS + Mevalonic Acid 22 SEP 00 28 SEP 00 10 OCT 00 DS 9 DS 15 DS 43, DG 0 3 11529- 11543 2 11529- 11543 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 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 Cholesterol 28 SEP 00 DS 15 2 11558 - 11571 Control 10 NOV 00 DG 18 2 10967 DS is used as an abbreviation for day o f study. DG is used as an abbreviation for day of (presumed) gestation. DL is used as an abbreviation for day of lactation. 000774 PROTOCOL 418-018: OHE 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 11020 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 92.7 68.8 80.5 124.9 137.9 91.9 90.1 99.1 109.3 96.1 108.4 73.9 80.0 103.0 114.2 77.7 87.7 73.9 78.5 100.2 1 1 1 .0 113.0 121.8 104.9 113.1 NO SURVIVING PUPS 90.7 102.4 93.1 105.0 107.8 133.0 140.5 100.5 90.1 153.8 96.2 115.9 116.3 87.8 122.4 94.4 81.0 123.9 125.4 117.5 111.0 116.3 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-270 c oo -i Cfl PROTOCOL 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 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 1 1 1 .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 45.1 119.1 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 88.1 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 41.4 131.1 116.9 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. 418-018:PAGE B-: oc o i to 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 3V (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 B 2 .6 70.4 43.9 NOT PREGNANT 75.6 77.9 33.2 NO SURVIVING PUPS NO SURVIVING PUPS 17.3 90.2 54.1 86.8 30.2 60.9 NO SURVIVING PUPS 67.0 8.6 51.6 20.3 66.8 40.1 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 PUPS 90.1 48.7 78.7 DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G) . POOLED LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. 418-018:PAGE B-272 000777 PROTOCOL 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 DATA - 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 c <*3 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 38 (PAGE 1): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP It 1 2 3 4 5 6 7 B 9 10 11 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 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 F A F A M A M A MA M A MA MA M S FA 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 F A FD 4 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 FM 5 FD 2 M A M A M A FA FA FA FA FA FA FA M A M A MA MA MA M A M A FA FA FA FA FS M A M A M A M A MA M A M A M A M A FA FA FA FA FA FA FS MA MA M A M A MA M A FA FA FA FA FA FA FA FA FA 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 FD 3 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 F A F A F A FM 2 M A MA MA M A MA M A M A M A M A FA FA FA FA FA FA MA MA M A M A MA M A M A M A M A FA M A M A M A M A M A MA M A MA FA FA FA FS FS M A M A FA FA FA FA FA FA MA MA MA M A M A MA M A MA M A FA FA FA FA FA FA M A M A MA M A M A M A MA FA FA FA FA FA FA FA 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 1 M A MA MA M A M A M A M A FA FA FA FA M A M A M A M A MA M A FA 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. 418-018:PAGE B-274 000779 PROTOCOL 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 2): 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 il 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER tt 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 MA MA MA MA MA MA MA MA MA MA MA FA FA FA MA MA M A MA M A MA MA M A FA FA FA FA FA MORIBUND SACRIFICED ON DAY 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT M A M A M A M A M A M A FA FA FA FA FA FA FA FA FA FA FA M A M A MA M A M A MA M A FA FA FA FA FA M A M A M A MA M A MA Ms FA FA FA FA FA FA FA FA FS MA MA MA M A M A MA FA FA FA FA FA FA FA FA FA MA MA 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 i MA MA M A M A M A MA 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 to A MA MA FA FA FA FA FA FA FA FA MA MA MA MA MA FA FA FA FA FA FA MA M A M A M A MA MA MA MA MA FA FA FA FA MA MA MA MA MA MA MA MA FA FA FA FA Fs MA MA MA MA MA MA MA FA FA FA FA MA M A M A M A MA MA M A 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. 418-018 :PAGE B-275 oo o *! or o PROTOCOL. 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 It 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 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 2 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 i 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 X 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 MA M A M A 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 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:PAGEB-276 000761 PROTOCOL 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 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 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 1 MD 2 MD 2 MD 2 MM 2 MM 2 MM 2 MM 2 MM 2 MM 2 MM 3 FM 2 FM 5 FM 3 MM 2 MD 1 MD 1 MD 1 MD 1 MD 1 FM 4 FM 3 FD 1 FD 1 FD 1 FD 1 FD 1 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 FD 2 MD 2 MD 2 MD 1 M S FD 2 FD 2 FD 2 FD 2 FM 2 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 M S FM 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 MD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 MM 2 MM 2 MM 2 MM 2 MD 1 FM 2 FM 2 FM 2 FD 2 FD 2 FD 1 FD 1 MD 1 MD 1 MD 1 MD 1 MD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 MM 5 MM 5 MM 2 MM 2 MM 2 MD 1 F A FM 4 FD 3 FM 3 FD 2 FD 2 FD 2 FM 2 FD 1 NOT PREGNANT MM 2 MM 2 MM 2 MM 2 MD 2 MD 2 FM 2 FD 2 FD 2 FD 1 FD 1 FD 1 FD 1 FD 1 MD 3 MD 3 MM 3 MD 2 MD 1 FM 4 FM 3 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 1 MM 3 MM 2 MD 1 MD 1 MD 1 MD 1 FM 2 FD 1 FD 1 FD 1 FD 1 FD 1 M A M A MD 3 MM 2 MD 1 MD 1 MD 1 MD 1 FD 4 FM 2 FD 1 FD 1 FD 1 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 FD 2 FD 2 FD 2 FD 2 MD 2 MD 2 MD 2 MD 2 MM 2 FD 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 1 FD 1 MD 3 MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 MD 1 MD 1 MD 1 MD 1 MD 1 MD 1 MD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 MD 1 MD 1 MD 1 MD 1 MD 1 MD 1 MD 1 FD 1 FD 1 FD 1 FD 1 FD 1 FD 1 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 c cr N PROTOCOL 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 5): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP It 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 K MATERNAL DOSAGE GROUP S CHOLESTEROL CONTROL 10957 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 M A MA FA FA FA FA FA FA FA FA FA MA MA M A MA MA M A MA FA FA FA FA FA FA FA FA MA M A MA MA MA M A M A MA FA FA FA FA FA FA FA MA M A MA MA MA MA MA M A 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 M A MA MA MA MA MA MA MA MA FA FA FA FA MA M A MA MA MA M A MA FA FA FA FA FA FA FA FA FA FA FA NOT PREGNANT MA MA MA MA M A MA MA M A 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 MA MA MA MA FA FA FA FA FA FA FA FA FA NOT PREGNANT MA MA MA MA MA 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 MA MA MA MA MA MA MA FA FA FA FA FA MA M A MA 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. 418-018:PAGE B-278 ooc >i 05 CJ 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 38 (PAGE 6): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 5 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS PUP It 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 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 i 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 1 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 i 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 i MA MA MA MA 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 1 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 1 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 1 M s FD 5 FM 3 FD 2 FD 2 FD 2 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-279 c vi CO PROTOCOL 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 7) : 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 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 1 MD i MD 1 MD 1 MD 1 FD 1 FD i FD i FD 1 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 FD 1 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 i 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 1 MD 1 MD 1 MD 1 FM 2 FD i FD 1 FD 1 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" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-280 c G? C/T PROTOCOL 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 # 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 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 MA MA MA MA MA MA MA M A M A M A FA FA FA FA FS MA M A MA M A FA FA FA FA FA FA FA FA FA FA MA MA MA MA MA MA MA MA MA MA FA FA FA FA FA MA MA 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 MD i 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 MA MA MA MA MS FA FA FA FA FA FA FA FA MA MA MA MA MA FA FA FA FA FA FA FA FA FA FA FA FA MA M A M A MA MA MA 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 M A MA MA MA MA FA FA FA FA FA FA FA FA FA FA MA M A M A M A MA MA 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 MA MA MA MA MA MA MA FA FA FA FA FA MA MA MA MA MA MA MA FA FA FA FA FA FA FA FA FA MA M A M A M A FA FA FA FA FA Fs MA MA MA MA MA MA MA MA MA FA FA FA FA FA MA MA MA MA MA MA MA M A 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 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-281 c GC a PROTOCOL 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 It 1 2 3 4 S 6 7 8 9 10 ii 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER 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 MA MA FA FA FA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA MA FA MA MA MA MA M A M A FA FA MA M A MA FA FA MA M A FA FA FA M A MM 2 M A F A F A M A M A MD 1 F A F A FA FA FA FA FA FA FA FA FA FA FA FA FA FA MA MA MA FA FA FA FA FA FA FA MA M A FA FA FA M A M A M A M A MM 3 M A M A MA FA FA MA M A FA FA FA M A M A M A MD 2 F A F A F A F A F A FD 4 MA FA 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 FA FA FA FA FA FA FA FA FA FA FA FA FA FA F A F A F A F A F A FD 2 FS 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. 418-018:PAGE B-282 000787 PROTOCOL 418-010: 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 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER It 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 MA MA MA MA MA MA MA MA FA FA FA FA FA FA FA MA M A M A MA MA 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 M A M A M A M A 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 MA MA MA MA MA MA MA M A MA 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 MA MA MA MA MA 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 FS 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 MA MA MA MA 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-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-283 o GO 00 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 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 il 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER 0 MATERNAL DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 11030 11031 11032 11033 11034 11035 1IQ36 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 1 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 1 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 M A 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 MA M A MA M A MA MA M A MA FA FA FA FA FA FA FA MA MA MA MA MA MA 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 M A MA M A MA MA M A 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-ALIVE, S-STILLBOEN, U-UNCERTAIN, D-DIED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-018:PAGE B-284 0007 C* C PROTOCOL 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 il 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER H 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 I 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 MA M A M A MA MA M A MA M A 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 X 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 X 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 CDi PROTOCOL 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 # 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 1 MD 1 F A F A F A F A F A F A MD 3 MM 2 MD 2 MD 1 MD 1 MD 1 MD 1 MD 1 FM 2 FD 2 FD 1 FD 1 FD 1 MD 1 MD 1 MD 1 MD 1 MD 1 MD 1 MD 1 MD 1 FM 2 FM 2 FD X FD 1 FD 1 FD 1 MD 1 MD 1 FD 1 FD 1 FD 1 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 1 MD 1 MD 1 MD 1 FD 2 FD 2 FD 1 FD 1 FD X 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 i MD X MD 1 MD 1 MD 1 M S FM 2 FM 2 FD 1 FD 1 FD 1 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-286 oo CNDi PROTOCOL 418-010: 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 TO 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 11542 2 1/11 PUPS: NO MILK IN STOMACH. 1 15/15 PUPS: NOT NURSING. 4 2 MG/KG PFOS + MEVALONIC ACID 10944 11547 4 1/ 1 PUP: NOT NURSING. 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. 418-018:PAGE B-287 000792 PROTOCOL 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 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 10967 3- 5 4- 5 1/11 PUPS: TIP OF TAIL BLACK.b 1/15 PUPS: RIGHT EYE, EXOPHTHALMOS.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 000793 PROTOCOL 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 3) : CLINICAL OBSERVATIONS FROM BIRTH TO DAY S 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. 115B7 11588 2 1/13 PUPS: NOT NURSING. 1 13/14 PUPS: NOT NURSING. 11589 11590 4 1/ 1 PUP: NOT NURSING. 1 15/16 PUPS: NOT NURSING. 2 5/15 PUPS: COLD TO TOUCH. 11592 X 15/15 PUPS: NOT NURSING. 11593 2 3 4- 5 1/12 PUPS: 1/ 7 PUPS: 1/ 6 PUPS: TAIL MISSING. TAIL MISSING. TAIL MISSING.b 11596 2 1/14 PUPS: COLD TO TOUCH. NOT NURSING. 11598 2 2/15 PUPS: 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 000794 PROTOCOL 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 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 1 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 BASE.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 000795 PROTOCOL 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 9) : 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 NURSING. 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-: co to 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 VEHICLE CONTROL 10906 5 10909 1 5 10911 4 5 10913 2 5 10914 5 11501 1 5 11502 1 5 C 11503 5 O 11504 3 M 5 11505 11506 11507 5 5 5 11508 1 5 11509 11510 11511 5 5 5 Complete necropsies were not performed on pups in whicl individual pup clinical observations table (Table 39) 1 ALL PUPS APPEARED NORMAL. 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. I PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 1 PUP: STILLBORN. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 2 PUPS: STILLBORN. TONGUE PROTRUDED. ALL OTHER TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 418-018:PAGE B-292 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 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 1 VEHICLE CONTROL (CONT.) 1X512 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 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11517 11518 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. o CA .P 11519 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. >i 11520 5 ALL PUPS APPEARED NORMAL. (ft 11521 5 ALL PUPS APPEARED NORMAL. r* 11523 5 ALL PUPS APPEARED NORMAL. 11524 5 ALL PUPS APPEARED NORMAL. 11525 5 ALL PUPS APPEARED NORMAL. 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. 418-018:PAGE B-293 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 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 000799 PROTOCOL 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 3 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. 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. 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 S 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. 008000 418-018:PAGE B-295 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 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. ALL 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 1 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 MILK 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. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 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 OOOOl PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6290.25) 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 1 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 MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11546 1 11 PUPS : FOUND DEAD. NO 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 MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 4 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11550 1 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 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 40 (PAGE 7): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS 08000 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. ALL 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 1 PUP: FOUND DEAD. NO MILK 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. ALL 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. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 10 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER CJ 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 PROTOCOL 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 8) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE LITTER NUMBER DAY POSTPARTUM OBSERVATIONS a 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. c o 11565 5 ALL PUPS APPEARED NORMAL. c 11567 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. Qt' 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. 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 PROTOCOL 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 6 1.6 MG/KG PFOS + CHOLESTEROL LITTER NUMBER 10971 DAY POSTPARTUM 1 2 OBSERVATIONS a 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 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. o ocr vi 10979 10983 10984 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. 3 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 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 PROTOCOL 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 6 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 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11576 5 ALL PUPS APPEARED NORMAL. oO' o tf) & o 11577 11578 11579 2 13 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 1 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 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. 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 PROTOCOL 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 e 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. ALL 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. I 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 c o cor PROTOCOL 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 MILK 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: STILLBORN. ALL TISSUES APPEARED NORMAL. 2 6 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL 3 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL 11587 2 12 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11588 1 1 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. rib ^ooo 418-018:PAGE B-303 PROTOCOL 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 B PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11591 1 1 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. Wo o 05 ' w CO 11593 11594 11596 1 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 2 15 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 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 PROTOCOL 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 7 2 MG/KG PFOS + CHOLESTEROL (CONT.) 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. oo 5 ALL PUPS APPEARED NORMAL. o 11004 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS Q 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 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 40 (PAGE 15): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST SUBSTANCE 8 0.4 MG/KG PFOS (CONT.) LITTER NUMBER 11600 DAY POSTPARTUM 1 5 OBSERVATIONS a 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 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. ALL TISSUES APPEARED NORMAL. 4 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11604 5 ALL PUPS APPEARED NORMAL. -s.'-' 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 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11012 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. TT9000 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 11016 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. 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. c CoO ro 11018 11611 11612 11613 11614 11615 11616 11617 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. 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. 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 PROTOCOL 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 POSTPA1 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. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11027 5 ALL PUPS APPEARED NORMAL. co 0HU5 11028 11620 11621 11622 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. ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 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. 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. 4I8-018:PAGE B-308 PROTOCOL. 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 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 MILK 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 S 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. oo 11032 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. AUTOLYSIS PRECLUDED FURTHER EVALUATION. o 5 ALL PUPS APPEARED NORMAL. P0 3 11033 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL. 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 PROTOCOL 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. ALL TISSUES APPEARED NORMAL. 5 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. ALL OTHER PUPS APPEARED NORMAL. o 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 PROTOCOL 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 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 i PUP: FOUND DEAD. NO MILK 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. O 11042 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. a ALL OTHER TISSUES APPEARED NORMAL. 2 11 PUPS: FOUND DEAD. NO MILK IN STOMACH. r-A K' ALL OTHER TISSUES APPEARED NORMAL. rUfc CD 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. 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 PROTOCOL 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. oo O ^ 1-* n! 11643 11644 11645 2 5 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. AITTOLYSIS PRECLUDED FURTHER EVALUATION. 5 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL OTHER PUPS APPEARED NORMAL. 1 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL OTHER PUPS APPEARED NORMAL. 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 PROTOCOL 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. 418-018:PAGE B-313 0ooc5 00 PROTOCOL 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 23): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS MATERNAL DOSAGE GROUP TEST S U B S T A N C E 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 11061 5 ALL PUPS APPEARED NORMAL. 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 II PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 11654 2 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 5 ALL PUPS APPEARED NORMAL. 11655 4 1 PUP: FOUND DBAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 11656 1 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 ALL PUPS APPEARED NORMAL. 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 necropBy. 418-018:PAGE B-314 o o o tis PROTOCOL 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 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. c r ig 11661 11662 11663 1 11 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 3 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 2 3 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 3 2 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 5 PUP APPEARED NORMAL. 1 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 2 13 PUPS: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 11664 1 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. 418-018:PAGE B-315 PROTOCOL 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 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 APPENDIX C PROTOCOL AND AMENDMENTS 000822 418-018:PAGE C-l O Primedica Argus Research Laboratories. Inc. 9Q5 Sheehv Drive. Building A Horsham. PA 1944 Telephone: (215) 443-8710 Telefax: (215)443-8587 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 00023 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 o f 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. 000824 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 H20). Supplier and lot identification of Tween 80 to be documented in the raw data. Reverse Osmosis Membrane Processed Deionized Water (RODI H20). 000S25 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 (-20 C). 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. 000526 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. G00S27 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. D IS P O S IT IO N : 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 Facility11'3'; 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. 000828 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. FI Generation: Pups will not be individually identified during lactation; all parameters will be evaluated in terms of the litter. 0000^9 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 o f Laboratory Animals(4). 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 6 4 F (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. 00030 418-018:PAGE C-9 Water: Protocol 418-018 Page 9 W ater 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. W ater 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. 000831 418-018 :PAGE C-10 F1 Generation Pups: Protocol 418-018 P age 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. 0Q0S32 418-018:PAGE C-ll Dosage Levels. Concentrations and Volumes: Protocol 418-018 P a g e 11 j 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" dosage Dosing Schedule 2" dosage Vi 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 3* dosage 5 hr 30 minutes after 2" RODI H ,0 500 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 (mL/kg) Argus Batch Number RODI H ,0 0 0 5 B-418-018-A(Dav.Month.Year! 0.5% Tween 80 0 0 5 B-418-018-B(Dav.Month.Year) Mevalonic Acid 1000 100 5- B-418-018-C(Dav.Month.Year) Cholesterol 500 100 5 B-418-018-D(Day.Month.Year) PFOS 0.4 0.08 5 B-418-018-E(Dav.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-H(Dav.Month.Year) PFOS 1.6 0.32 5 B-418-01 B-l(Dav.Month.Year) PFOS 2 0 40 5 B-418-018-J(Day.Month.Year) The test ana control articles will be considered 100% pure tor tne purpose ot dosage calculations. ' Mevalonic acid will be dosed twice daily. 000533 418-018:PAGE C-12 TESTS. ANALYSES AND MEASUREMENTS - Fo GENERATION: Protocol 418-018 P ag e 12 Viability: All Periods: At least twice daily. Clinical Observations and/or General Appearance: Acclimation Period: Dosage Period: Day of Sacrifice: Maternal Behavior: At least once. 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). Once. 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: Sacrifice: Weekly to cohabitation. Daily during presumed gestation and on Day 1 postpartum (rats assigned to natural delivery). 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. 000634 418-018:PAGE C-13 Mating Performance: Protocol 418-018 P age 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.) 000S35 418-018:PAGE C-14 Fetal Observations: Protocol 418-018 P ag e 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. 000836 418-018:PAGE C-15 Protocol 418-018 P ag e 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. 000837 418-018:PAGE C-16 NECROPSY - Fo GENERATION: Protocol 418-018 P ag e 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-Sectionino: 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. 000838 418-018:PAGE C-17 Protocol 418-018 P ag e 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 (s-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 (s-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 (s-70C ) until shipment for possible analysis. The median liver lobe will be frozen and stored (s-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. 000S39 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. 000^40 418-018: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 autolysis), 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. 000841 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 Puds 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. 000842 418-018:PAGE C-21 Protocol 418-018 P a g e 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 (s-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. 000S43 418-018 .'PAGE C-22 PROPOSED STATISTICAL METHODS*6'12': 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 II. Nonparametric0 A. Bartlett's Test0 A. Kruskal-Wallis Test ($75^0 ties) Significant at p<0.001 Not Significant Significant at p<;0.05 Not Significant Nonparametric Analysis of Variance Dunn's Test Significant at psO.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. 000844 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. W ater and Bedding Analyses. Packing and/or Shipment Lists. 000S45 418-018:PAGE C-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. 000846 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 7, 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 Crl:CDBR 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 o f Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski, E. (1964). Frbemethode 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. OOOS47 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 rrz) C ______________ Dena C. Lebo, V.M.D. Chairperson, institutional Animal Care and Use Committee FOR THE SPONSOR J l Deanna J. Lueb Study Monitor John Butenhoff, Ph.D., DABT, CIH Alternate Study Monitor S 7 s & / s cc<\ Date 2 3- At/fr - Z/P&O Date 000848 418-018:PAGE C-27 ATTACHMENT 1 STUDY SCHEDULE 000&49 ATTACHMENT 1 418-018:PAGE C-28 Protocol 418-018 P ag e 1 of 2 SCHEDULE3 REPLICATE 1b 22 AUG 00 Animal Receipt - Acclimation Begins (Female Rats). 29 AUG 0 0 - 12 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 -1 6 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 O CT 00 23 OCT 00 First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. 31 O CT 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 ATTACHM ENT 1). Q00S5Q 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 O CT 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. 000L51 418-018.-PAGE C-30 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 000852 418-018-.PAGE C-31 DATA SHEET 3M Center St. Paul, Minnesota 55144-1000 1-800-364-3577 or (612) 737-6501 (24 hours) Copyright, 1998, Minnesota Mining and Manufacturing Coapany. All rights reserved. Copying and/or downloading of this inforaation for the purpose of properly utilizing 3M products is allowed provided that: 1) the inforaation is copied in full with no changes unless prior agreeaent is obtained froa 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: PC-95 FLUORAD 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 ZF-0002-1044-1 ISSUED: January 29, 1998 SUPERSEDES: Noveaber 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=l (Bulk) 0S 7 -8 (0 .1% Aqueous) N/D N/D APPEARANCE AND ODOR: Light colored, free flowing powder. 000S53 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately USDS: F C -95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 3. FIRE AMD EXPLOSION HAZARD DATA 418-018:PAGE C-32 PAGE 2 FLASH POINT:................... None FLAMMABLE LIMITS - LEL:.. N/A FLAMMABLE LIMITS - UEL:.. N/A AUTOIGNITION TEMPERATURE:..... N/A EXTINGUISHING MEDIA: Water, Carbon dioxide, Dry chemical, Foaa SPECIAL FIRE FIGHTING PROCEDURES: Wear full protective clothing, including helaet, self-contained, positive pressure or pressure deaand breathing apparatus, bunker coat and pants, bands around aras, waist and legs, face aask, 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 0006S4 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately MSDS: PC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 5. ENVIRONMENTAL INFORMATION (continued) 418-018:PAGE C-33 PAGE 3 accept cheaical waste. ENVIRONMENTAL DATA: 96-Hr. Aquatic Fish LC50, Fathead Minnow(Pimephales proaelas)=38 mg/1, Bluegill Sunfish(Lepoais macrochirus)*68 ag/1, Rainbow Trout(Salmo gairdneri)11 ag/1; 48-Hr. EC50, Daphnia Magna * 50 ag/1; COD=.004 q /q ; BQD20 - Nil. REGULATORY INFORMATION: Volatile Organic Coapounds: N/A. VOC Less H20 & Exeapt Solvents: N/A. Since regulations vary, consult applicable regulations or authorities before disposal. U.S. EPA Hazardous Haste Number * None (Not U.S. EPA Hazardous). This product complies with the cheaical registration requirements of TSCA, EINECS, COSL, AICS, MITI and Korea. EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes 6. SUGGESTED FIRST AID EYE CONTACT: Inmediately 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. Hash contaminated clothing before reuse. INHALATION: If signs/symptoms occur, remove person to fresh air. If signs/symptoms continue, call a physician. IF SWALLOHED: Drink two glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION EYE PROTECTION: Avoid eye contact. Wear vented goggles. 000655 AbDreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 3DS: FC*95 FLUORAD Brand Fluorocheaical Surfactant anuary 29, 1998 7. PRECAUTIONARY INFORMATION (continued) 418-018:PAGE C-34 PAGE 4 KIN PROTECTION: Avoid skin contact. Hear appropriate gloves when handling this Material. A pair of gloves Made froa the following Material(s) are reconoended: butyl rubber. Use one or more of the following personal protection iteas as necessary to prevent skin contact: head covering, coveralls. Protective garaents (other than gloves) should be aade of either of the following Materials: polyethylene/polyvinylidene chloride (Saranex). lECOMMENDED VENTILATION: Use with appropriate local exhaust ventilation. Use in a wellventilated area. Provide sufficient ventilation to maintain eaissions below recommended exposure limits. If exhaust ventilation is not adequate, use appropriate respiratory protection. IESPIRATORY 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. Mash 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 O Q Q l.S S SOS: FC-95 FLUORAD Brand Fluorocheaical Surfactant anuary 29, 1998 EXPOSURE LIMITS (continued) 418-018:PAGE C-35 PAGE 5 NGREDIENT VALUE UNIT TYPE AUTH SKIN* OTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 TWA 3M Y SKIN NOTATION: Listed substances indicated with 'Y' under SKIN refer to he potential contribution to the overall exposure by the cutaneous route .ncluding mucous aeabrane and eye, either by airborne or, acre particularly, >y direct contact with the substance. Vehicles can alter skin absorption. SOURCE OF EXPOSURE LIMIT DATA: 3M: 3M Recommended Exposure Guidelines 8. HEALTH HAZARD DATA IYE CONTACT: Mild Eye Irritation: signs/symptoms can include redness, swelling, pain, and tearing. SKIN CONTACT: Mild Skin Irritation (after prolonged or repeated contact): signs/symptoms can include redness, swelling, and itching. May be absorbed through the skin and persist in the body for an extended time. INHALATION: May be harmful if inhaled. May be absorbed by inhalation and persist in the body for an extended time. Single overexposure, above recommended 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. Hay be harmful if swallowed. MUTAGENICITY: Hutagenicity assays indicate the product is not mutagenic. 000&57 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately HSDS: PC-95 FLUORAD Brand Fluorocheaical 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, ainformation obtained from database may not be as current as the 0G0S58 information in the MSDS available directly from 3M. *w fcwbw >*-*w 418-018:PAGE C-37 AMK OU^F M C T O - i)^AJ 3050 Sbtuc Slmst PRODUCT t: C8503 NAME: CHOXtESTEROIfittmLouis. MtcsounS9103 USA 'A SHEET, Valid 8/2000 - TbBphBrtkBttWZi-SSSSMaiAini-s? d 07/25/2000 15:03 F*x800-325-S052* (314)771-5757 emsil: no-atd8s>ai.eBn- i.iigms-iioricfl.eom 3050 Spruce Street ;t. Louis, MO 63103, US Technical Phone: 314 771 5765 Emergency Phone: 314 771 5765 rax: 800 325 5052 fftM -t d f o A - 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-BETA-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 SREATHING GIVE ARTIFICIAL RESPIRATION. 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. nftr SECTION 8. - - - - - - EXPOSURE CONTROLS/PERSONAL PROTECTION- - - - - A / t l U C S " CHEMICAL SAFETY GOGGLES. COMPATIBLE CHEMICAL-RESISTANT GLOVES. NIOSH/MSHA-APPROVED RESPIRATOR. Page 1 't't art c a mmi t t t d to the S u c c c u 0/ our C m i o m c r i , Empt v j t and 418-018:PAGE C-38 PRODUCT I ; C 8503 3050 Spruce street NAME: CHOLESTEROI^mLow*.Missouri63103USA SAHBCY AAffA SHEET, V a l i d 8 / 2 0 0 0 - fc0rfaft<Mk52i-a956*(3i)77''-576S d 07/25/2000 15:03 ____ _________________ FaxM M C S* 314)77'.5757 mart- wg.alrtgfial m m _______ www.aigma-aidnen.eem SHOWER ANp 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. CTION 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 1 0 . ---- --------- -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 000.60 IARC CANCER REVIEW :ANIMAL INADEQUATE EVIDENCE IMEMDT 10, 99,1976 Page 2 ^ < re committed to * i.ct.ji { ou, Emalejtfi ni *1 ' ' T.rknolo,, try 5 .,,iff. - w--6^ 418-018:PAGE C-39 rnwrgp PRODUCT k BFrrM .wmw 9Uau apniua #: C8503 NAME: CHOLESTEROlBamtLou.M*eun63103USA SHEET, Valid 8/2000 - TMiNl>MP-52i-6956 (3U) T71-5765 07/25/2000 15:03 R * O M a M . 0 U ) 771-5757 mil: cta-ldaai.corn THanr.QTiaTF. p.UTngwrr---- TMEMnX-11 -9^. | yy^ttgma-Aiancn.eom IARC CANCER REVIEW .-GROUP 3 IMSUDL 7,161,1987 NOHS 1974: H2D 80865; NIS 8; TNF 1321; NOS 10; TNE 13980 NOES 1983: KZD 80865; NIS 5; TNF 4091; NOS 11; TNE 41411; TFE 30220 EPA GENETOX PROGRAM 1988, 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 S2GMA-ALDRICH CO. LICENSE GRANTED TO MAKE UNLIMITED PAPER COPIES FOR INTERNAL USE ONLY 000S61 Page 3 V t a r c c o m m i , ; t o t k c S v c c c x j o / 0 u r C u c t e m c r s . F . n k t e j t r* and X h a r c h o l Jt'i thfuojfli Li/c Science. Teelinoloj, and I t t v i t * . 418-018:PAGE C-40 J SIG 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 S00 325 5052 Emergency Phone: 414 273 3850 Ext.5996 Section 2 Composition/lnformation on Ingredient Substance Name DL-MEVALONIC ACID LACTONE Formula Synonyms C6H1003 Section 3 - Hazards Identification CAS # 674-26-0 SARA 313 No For additional information on toxicity, please refer to Section 1 1 . Section 4 First Aid Measures Oral Exposure If swallowed, wash out mouth wrth water provided person is conscious. Call a physician. Inhalation Exposure 11 inhaled, remove to fresh air. Hnot breathing give artificial respiration. If breathing is difficult, give oxygen. Dermal Exposure In case of contact, immediately w ash skin with soap and copious amounts of water. Eye Exposure In case of contact, immediately flush eyes with co p e u s am ounts of water for at least 15 minutes. Section 5 - Fire Fighting Measures Flash Point: 235 *F 113 *C Autoignition Temp: N/A Flammability: Extinguishing Media Suitable Water soray. Carpon oioxide. dry chemical powder, or appropriate foam. N/A 000862 418-018:PAGE C-41 Firefighting Protective Equipment W ear self-contained breathing apparatus and protective clothing to prevent contact with skin and eyes. Section 6 - Accidental Release Measures Procedurefs) of Personal Precaution^) W e a r respirator, chemical safety goggles, rubber boots, and heavy rubber gloves. Methods tor Cleaning Up Absorb on sand or vermiculite and place in closed containers tor disposal. Ventilate area and wash spill site alter matenal pickup b complete. Section 7 Handling and Storage Handing 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/MSHA-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 Physieal/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 TernDorature or Priro 5 mmHg 000863 Sigma Chemical - M4667 Page 2 Sigme-Aldrich Corporation rwin.eemo-oMhch.com 418-018:PAGE C-42 V olatile* VOC Content Water Content Solvent Content Evaporation Rate Viscoaity Partition Coefficient Decomposition Temp. Flesh Point *F Flesh 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 Protea from moisture. Conditions to Avoid Protea from moisture. Materials to Avoid Strong oxidizing agents. Hazardous Decomposition Products Hazardous Decomposition Product* 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 irntating to mucous membranes and upper respiratory tract. M ultiple Routes May be harmlul 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 with 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 Sigm a-Aldrich Corporation <"*n-agin*-aidncti.eom 000564 418-018:PAGE C-43 Non-Hazardous for Transport This substance is considered to be non-hazardous tor transport. IATA Proper Shipping Name: None Non-Hazsrdous for Air Transport Non-hazardous for air transport Section 15 - Regulatory information Unifad States Regulatory information SARA 313 Usted: No Section 16 - Other Information Warranty The above information s 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 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. Sigma Chemical - MA667 Page 4 Sigm e-Aldrich Corporation nrw.agme-aWheh.cmi G00S65 418-018:PAGE C-44 ATTACHMENT 3 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE 000866 418-018:PAGE C-45 ATTACHMENT3 Protocol 418-018 Version: 418-018(16 AUG001 Page 1of 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 W ater 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 0G0S67 418-018:PAGE C-46 ATTACHMENT 3 Protocol 418-018 Version: 418-018(16 AUG00) 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 50C = 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 TE ST 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 80C - 5 C 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 TE ST 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 000868 418-018:PAGE C-47 ATTACHMENT 3 Protocol 418-018 Version: 418-018(16 AUG00) 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 TE ST 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 TE ST 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 TE ST 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 000869 418-018:PAGE C-48 ATTACHMENT3 Protocol 418-018 Version: 418-018(16 AUG001 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 TE ST 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). 0QQS70 418-018:PAGE C-4'9 ATTACHMENT3 Protocol 418*018 Version: 418*018(16 AUG001 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 by: . Date: ^ i-^ o c -rc o Clarification: Y eW S ee attached clarification form.) Initials/Date : X t/co 000871 418-018:PAGE C-50 ATTACHMENT 4 TISSUE COLLECTION AND PROCESSING 000872 418-018-.PAGE C-51 ATTACHMENT 4 Protocol 418-018 P a g e 1 of 1 Tissue Collection and Processing Sample Type Units Day 21 Gestation Fetuses Blood Pooled/litter Aliquots 1/2plasma 1/4 serum 1/4 serum Storage/Shipping Conditions <-20 <-20 <-20 Liver Pooled/litter 1liver 1/3 all livers 1/3 all livers Gluteraldehvde. RT <-20 <-20 Dav 21 Gestation Dams Blood 4 mL/dam 1/3 all livers 1/2p la s m a 1/4 serum 1/4 serum Liquid nitrogen. <-70 <-20 <-20 <-20 Liver each dam right lateral lobe median lobe 0.3-0.5 cm section remainder Liquid nitrogen. <-70 <-20 Gluteraldehyde. RT r ro 1 VI VI Day 5 Lactation Dams Milk 100 pL/dam Blood 4 mL/dam 1/2plasma 1/4 serum 1/4 serum <-20 <-20 <-20 Liver each dam right lateral lobe median lobe 0.3-0.5 cm section Remainder Liquid nitrogen. <-70 <-20 Gluteraldehyde. RT <-20 Heart each dam Gluteraldehyde. RT Dav 5 Postpartum Pups Blood Pooled/sex/litter 112 plasma 1/4 serum 1/4 serum <-20 <-20 <-20 Liver Pooled/sex/liiter 1/3 all livers 1/3 all livers <-20 <-20 1/3 all livers Liquid nitrogen. <-70 Hearts 2male. 2 female, per Gluteraldehyde. RT sex per litter PWL = Pnmedica Worcester Laboratories Tot chol = Total cholesterol RT = Room Temperature EM = Electron Microscopy Shipped to Analvsis PWL 3M AniLytics ?M 3M AniLytics 3M Mevalonic acid PFOS LDL. HDL. tot chol. triel Possible EM PFOS LDL. HDL. tot chol. trisl Possible Biochemical PWL 3M AniLytics 3M 3M 3M Mevalonic acid PFOS LDL. HDL. tot chol. trielv Possible Biochemical PFOS Possible EM Ant Lyncs LDL. HDL. tot chol. trielv AniLytics PWL 3M Am Lvtics 3M 3M 3M Total cholesterol Mevaiomc acid PFOS LDL. HDL. tot chol. triel Possible Biochemical PFOS Possible EM Am Lvtics 3M LDL. HDL. tot chol. trielv Possible EM & light microscopy PWL Mevalonic acid 3M PFOS AniLytics LDL. HDL. tot chol. triel 3M PFOS AniLytics LDL. HDL. tot chol. triel 3M Possible Biochemical 3M Possible EM & light rrucroscopv trigl = Triglyceride 000873 OPwmedica. 418-018:PAGE C-52 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 000874 Any revisions made to this finalized amendment must be made by subsequent amendment. Reason for Change: 418-018:PAGE C-53 Protocol 418-018 Amendment 1 Page 2 M )gna C. Lebo, V.M.D. Date Deanna J. LuebKer, M.S. Chairperson, Institutional Animal Care Study Monitor and Use Committee Date John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor Any revisions made to this finalized amendment must be made by subsequent amendment. 000875 O Primedica 418-018:PAGE C-54 Argus Research Laboratories. Inc. 90S 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 - 2 2 November 2000 1. Alternate Study Monitor (page 1 of the protocol): The telephone number for John Butenhoff. Ph.D.. DABT, CIH 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. T4 and TSH levels, LDL, HDL and T .triglycerides. The first priority will be analysis for total cholesterol and glucose. The second priority will be analysis for total and free 3 T4 and TSH levels. The third priority will be analysis for LDL. HDL and triglycerides. 000376 Any revisions to this finalized amendment must be made by subsequent amendment 418-018.-PAGE C-55 The liver and serum samples will be shipped to: Protocol 418-018 Amendment 2 Page 2 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 1 mL 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 pL) 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 T3,T4 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 (<-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 000877 Any revisions to this finalized am endment 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 Puds (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. __ ueg xjeorge. Dearlove, Ph.D., DABT Associate Director of Research 3 Date Ryftond G. Associate Directo) Study Director ., DABT Date search Chairperson, Institutional Animal Care and Use Committee jdjLJG L& ^J2i Deanna J. Lueb'ker, M.S. Study Monitor J . i ` /? * '/? < * * * John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor 000878 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 11), 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 o f the above analyses. Remaining samples will be retained at Primedica Worcester Laboratories or the Testing Facility until notification from the Sponsor for disposition. 00087 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, T4 and 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 Tj, 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. Associate Dire Study Director Dena C. Lebo, V.M.D. Date Deanna J. Luet Chairperson, Institutional Animal Care Study Monitor and Use Committee J- John Butenhoff, Ph.D., DABT, CIH Alternate Study Monitor Date J ? fr/ 0 / dM j/A / Date 000S80 Any revisions made to this finalized amendment must be made by subsequent amendment. OPmmedica 418-018:PAGE C-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 Pups (page 10 of the protocol): [Effective Date: 21 August 2000] Day 1 o f lactation (postpartum) is defined as the day o f birth and is also the first day on which all o f 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 F 1 Generation, Body Weights. 2. Scheduled Sacrifice - FI Generation Pups (page 20, 21, page 1 o f ATTACHMENT 4 and item 5 o f 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 o f 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 o f one male and one female F 1 generation pup from each o f 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 000881 Any revisions m ade to this finalized am endm ent must be m ade by subsequent am endm ent. Reason for Change: This change was made at the request o f the Sponsor. 418-018:PAGE C-60 Protocol 418-018 Amendment 4 Page 2 Gewge . Dearlove, Ph.D., DABT Associate Director o f Research Date Associate Director oTResearch and Study Director Date _______________________ . leresa H. WoodArd, D.V.M. Date Chairperson, Institutional Animal Care and Use Committee Study Monitor y w i--1. 7 ~. John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor Date 0008S2 Any revisions made to this finalized amendment must be made by subsequent amendment. 418-018:PAGE C-61 90S S h tc iif Drive, Bldg. A Horsham, PA 19044 Telephone;(215) 443-87(0 Tekfx (215)443-9587 ARGUS RESEARCH Charles River Laboratories Discovery and Developm ent 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 o f 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 o f 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 o f 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. 0008S3 Any revisions to this finalized am endm ent must be made by subsequent am endm ent Reason for Change: This change was made at the request o f the Sponsor. 418-018:PAGE C-62 Protocol 418-018 Amendments Page 2 -George E. Dearlove, Ph.D., DABT Associate Director o f Research Date 'OEjZ& ZO Zz- Raymond G. fork, Ph.D., DABT Date Associate Direct f Research and Study Director [ j - (. r d u + .J Hpbti- Theresa H. W oodard, D.V.M. Date Deanna J. Luel er, M.S. Chairperson, Institutional Animal Care Study Monitor and Use Committee . / S fc O O ? Date o7-/0 John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor 0008S4 Any revisions to this finalized amendment must be made by subsequent amendment. 418-018:PAGE C-63 905 SheehyDrive,BMg.A Horsham,PA 19044 Telephone:(215) 443-6710 Tekfioc (2IS)443-8587 ARGUS RESEARCH Charles Hiver Laboratories Discovery and Developm ent 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 o f 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 o f 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. 000885 Any revisions to this finalized amendment must be made by subsequent amendment. 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. 4 )0 1a fa - jeor^p E. Dearlove, Ph.D., DABT , Associate Director o f Research Date G. York, AssocWe Director and Study Director c. Dena C. Lebo, V.M.D. Member, Institutional Animal Care and Use Committee Date Deanna J. Lueb! Study Monitor /O i t / o ; John Butenhoff, Ph.D., DABT, CIH Date Alternate Study Monitor Unlu/oZK Date 000886 Any revisions to this finalized amendment must be made by subsequent amendment 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 C om puter 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 H PLC Column:Genesis C8 (Jones Chromatography), 2.1 mm x 50 mm, 4p Column Temp.: 35 C Injection Voi.: 10 pL M obile Phase (A): 2 mM Ammonium Acetate in ASTM type I water M obile Phase (B): Methanol Flow Rate: 0.3 mlVmin. 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 C[g etc.) can be used, provided equivalent chromatography is obtained. Ions monitored: Analyte PFOS Mode Transition Monitored negative 499 99 Approximate Retention Time (min) 4.40 000887 X -305 8 Research Drive State College, PA 16801, USA YT: 800.281.3279 F; 814.272.1019 Btygen.com 418-018:PAGE C-66 Proven Results. On a day-to-day basis, the retention times may vary slightly depending un 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 pL) of each calibration standard (prepared in MeOH) into the LC/MS/MS. b. Use linear, 1/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 resflA ^hll outside these GQ0888 3 0 5 8 Research Drive State College, PA 16801, USA T: 8 0 0 .2 8 1 3 2 1 9 F: 814,272.1019 exygen.com RES E ARC H 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 piL) 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. 00889 ygen.com APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 00090 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 INCDATE SPEC. COLLECTED: 11654 DATE SPEC. RECEIVED: RAT DATE REPORTED: 418018 PUPS LD5 AGE: 13 SEX: B 0035745 02/ 02/2001 02/13/2001 RESULTS UNITS REFERENCE: RANGE ARGUS CHEM PROFILE CHOLESTEROL-----GLUCOSE LDL-DIRECT h d l -d i r e c t 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 REPORT 000891 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 O C T 00 21 O C T 00 01 NOV 00 17 N O V 00 02 SEP 00 28 SEP 00 11 O C T 00 12 O CT 00 21 OCT 00 01 NOV 00 01 NOV 00 02 NOV 00 17 N O V 00 Day o f Study DS 8 D S 15 DS 24 DS 44 DG 8 DG 20, DL 2 DG 21, DL 2 DS 5 D S 15 DS 44, DG 0 DS 45, DG 0 D G s 9, 11 D G 21 DG 6 DS 50, DGs 4, 5, f DL 1 Dosage Number A dm inistered Early or Late 2 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 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 o f 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 1 mg/kg PFOS Dosage Not A dm inistered Cholesterol PFOS Date 28 SEP 00 01 O C T 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. 000S92 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 Identification 1.6 mg/kg PFOS 13 2 m g/kg PFO S Date 01 NOV 00 07 NOV 00 07 NOV 00 Day o f Study DS 65 D G 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. 000893 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 + M evalonic Acid 2 mg/kg PFOS + M evalonic 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 -1 1 5 1 2 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 m g/kg PFOS + M evalonic Acid 2 mg/kg PFOS + M evalonic 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 m g/kg PFO S + M evalonic Acid Date 04 NOV 00 04 NOV 00 19 N OV 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. 000894 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 m g/kg PFOS + M evalonic 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 D G 21 D G 21 DL5 DL5 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 o f the following dams. Dosage Group 5 8 Identification Cholesterol Control 0.4 mg/kg PFOS Date 22 NOV 00 22 NOV 00 Day o f 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. 000S95 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 + M evalonic Acid Date 25 NOV 00 25 NOV 00 Day o f Study DL5 DL5 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. v Raytnoiicl G. York, Ph. Associate Director of Resi and Study Director /(? b e d 1Q Z . Date 000896 APPENDIX E CERTIFICATE OF ANALYSIS 000897 418-018:PAGE E-l Certificate Of Analysis FC-95, Lot217 M arch 9, 2000 Richard M. Payfer This sample was analyzed using LC/MS, lH-NMR, lJF-NMR, and elemental analyses techniques. The results o fthese tests show due sample to contain the following weight percent composition: CaFsSOsTT cjFissnr GtfnSOsT ^i3sosir CtFwSOs-K" CsFnSOsTT CsFisSOsTT CjoFsiSOalT uFaSbjT^ CFrCda'lT SFsCsFj^l^ 0.04% 03% 131% uo%. 53t% U93S 90.49% 539*5 OIF55 53455 0.0S% 035% Additionally, the isomer distribution o f the sample was determined using ,9F-NMR techniques and found to contain foe following mole percent composition: CF3(CF2V SQ 3-K (Normal chain, where x is mainly 7) CF3(CF2VCF(CF3HCF2V SCb" K7 (Internal monometbyl branch, where x+y is mainly 5, and x * 0, y * 0) (CFs^CFKCFjV S O j-K 1 (Isopropyl branch, where x is mainly 5) CxF2*t i-C F (C F 3)-S O j'K ' (Alpha branch, where x is mainly 6) (CF3)3O (C F2V S 0 3'K t (trbutyl branch, where x is mainly 4) CF3-((jF2)x-C(CF3M CF2V SO3' (Internal gem-dimethyl branch, where x+y is mainly 4, and x * 0) 70.5% 17.1% 103% 1.6% 0.2% 0.2% 000898 418-018:PAGE E-2 SIG M A 3050 Sprue* Straat SaintLouis.Mrssoun63103 USA Taiaphon* 600-521-6956 (314) 771-5765 Fax 800-325-5062(314)771-5757 mail:ig-aldOsiaJ.com wwwjigma-aidhcti.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 SIGHA-ALDR1CH, INC. WARRANTS THAT ITS PRODUCTS 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 MAT APPLY. PLEASE SEE REVERSE SIDE OF THE INVOICE OR PACKING SLIP. SIGMA BRAND PROOUCTS ARE SOLD EXCLUSIVELY THROUGH SIGMA-ALDRICH, INC. 000899 W t a r c c o T n m i f t * ^ 10 *^ * S w e c e j j o f o u r C w s i o m e n . and APPENDIX F HOMOGENEITY AND CONCENTRATION ANALYSIS REPORT 000900 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 PROTECT Study Protocol Number: 418-018 Exygen Study Number: 023-069 Total Pages: 21 000901 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. mny R. Dicker Principal Investigator Exygen Research Date X L .'U'LYIU Deanna J. Luebker,\JW.S. Sponsor Representative 3M Corporate Toxicology Exygen Research i l / c n frQ D O ^ Date 000902 Page 2 of 21 418-018:PAGE F-3 Exygen Study N o.: 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. P ro to c o l R e v ie w 2. Extraction, F o rtific a tio n 3. Raw D ata R eview 4. Draft Report R ev iew 5. Final R eport R ev iew D ate InsDected D ate R eported to Exygen M anagement D ate Reported to Study D irector and SDonsor M anagem ent 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 A *-- ___ A ngela' M organ Q u ality A ssu ran ce A uditor / & fox- D ate Exygen Research 000903 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: --jS-IPtduY______ Emily R. Erecker Scientist Exygen Research tol&irL__ Date Exygen Research Facility Management: 30-C1T-O7Date Study Director, Argus Research: Sponsor Representative, 3M: 3M Corporate Toxicology Exygen Research /y-zUoL' -OZ. Date Date 000904 Page 4 of 2 1 418-018:PAGE F-5 E xygen Study N o.: 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 000905 Page 5 of 21 418-018:PAGE F-6 Exygen Study N o.: 023-069 PRO.IECT 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 000906 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.................................................................. 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 11 Exygen Research 000907 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 PFO S... 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 000908 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. 000309 Exygen Research 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 PFOS Exveen Control No. 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 (C&FnS03~) O CsF-t/S------O II o Note: The neutral molecule and standard form from which PFOS (anion) is derived, is potassium perfluorooctanesulfonate [C8F 17SO3K], 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 S tandards 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. 000910 Exygen Research Page 10 of 21 418-018:PAGE F -ll 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 000911 Exygen Research Page 11 of 21 418-018:PAGE F-12 Exygen Study No.: 023-069 HPLC CoIutmv.Genesis Cg (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: Analyte 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 000912 Page 12 of 21 418-018:PAGE F-13 ExygenStudyNo.: 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 ue 1000 ng Where: DF = Dilution Factor Equation 3 calculated the percent recovery. Equation 3: Recovery (%) = (anal, found (ue/mL) 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) = 1380 - 41.35451 721.773 = 1.85 ng/mL Exygen Research Page 13 of 21 418-018:PAGE F-14 Exygen Study No.: 023-069 From equation 2: Analyte found (pg/mL) = (1.85 ng/mL x 200,000) x 1 fig 1000 ng = 370 |ig/mL From equation 3: % Recovery = 370 ug/mL x 100 400 ng/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 000914 Page 14 of 21 418-018:PAGE F-15 Exygen StudyNo.: 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 000915 Page 15 of 21 418-018:PAGE F-16 Exygen Study No.: 023-069 TABLES Exygen Research 000916 Page 16 of 21 418-018:PAGE F-17 Exygen StudyNo.: 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 of 6M (0.4 mg/mL) 28.AUG.00 5 o f 6B (0.4 mg/mL) 28.AUG.00 1 of 2 ( 0 mg/mL) 18.SEP.00 1 of 2 (0.08 mg/mL) 18.SEP.00 1 of 2 (0.16 mg/mL) 18.SEP.00 1 of 2 (0.20 mg/mL) 18.SEP.00 1 o f 2 (0.24 mg/mL) 18.SEP.00 1 of 2 (0.32 mg/mL) 18.SEP.00 1 o f 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 of 2 (0 mg/mL) Ol.NOV.OO 1 of 2 (0.08 mg/mL) Ol.NOV.OO 1 of 2 (0.16 mg/mL) Ol.NOV.OO 1of 2 (0.20 mg/mL) Ol.NOV.OO 1 of 2 (0.24 mg/mL) Ol.NOV.OO 1 of 2 (0.32 mg/mL) Ol.NOV.OO 1 of 2 (0.40 mg/mL) Ol.NOV.OO 1 of 2 (0 mg/mL) 15.NOV.OO 1 of 2 (0.08 mg/mL) 15.NOV.OO 1 of 2 (0.16 mg/mL) 15.NOV.OO 1 of 2 (0.20 mg/mL) 15.NOV.OO 1 of 2 (0.24 mg/mL) 15.NOV.OO 1 of 2 (0.32 mg/mL) 15.NOV.OO 1 of 2 (0.40 mg/mL) 15.NOV.OO 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 Analyte Peak Found Area (ng/mL) 1380 1.85 1522 2.05 1551 2.09 ND ND 995 1.32 2156 2.93 714 0.932 867 1.14 1079 1.44 1385 1.86 1495 2.01 1423 1.91 1313 1.76 91 NQ 1002 1.33 2046 2.78 743 0.972 863 1.14 1054 1.40 1332 1.79 98 NQ 1001 1.33 2082 2.83 802 1.05 826 1.09 1145 1.53 1481 1.99 Analyte Found (Mg/mL) 371 410 418 ND 66.1 146 186 229 288 372 403 383 352 NQ 66.5 139 194 228 281 358 NQ 66.5 141 211 217 306 399 Sample Cone. (Mg/mL) 400 400 400 - 80 160 200 240 320 400 400 400 400 80 160 200 240 320 400 - 80 160 200 240 320 400 Ree. (%) 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 000917 Page 17 of 21 418-018:PAGE F-18 Exygen Study No.: 023-069 FIGURES Exygen Research 000918 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 000919 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-1M Sm (Mn, 3x2) 15-Feb-200214:35:32 LC/MS/MS #7 MRM o f 1 Channel ES- Exygen Research 000920 Page 20 of 21 418-018:PAGE F-21 Exygen Study No.: 023-069 Figure 3. Chromatogram Representing Dosing Solution Sample (Exygen ID: 0200283, Set: 021402A) 0200283, DFs 200,000 021402A-109 Sm {Mn. 3x2) 100~i 15-f=et>-200215:39:02 LC/MS/MS #7 4.96 1380 MRM o f 1 Channel E5499 > 99 9.36e3 Area - ........ .......i* .,n ............................ 1.00 1.50 2.00 2.50 3.50 4.00 Time 6.00 6.50 7.00 7.50 8.00 Exygen Research *>00921 Page 21 of 21 APPENDIX G TEMEPRATURE AND RELATIVE HUMIDITY REPORTS 000922 ARGUS 418-018:PAGE G-l Tem perature and Relative Hum idity Report Location: Room 17 Protocol N um ber 418018 Range o f Dates: 22-Aug-2000 14:44 to 12-Nov-2000 08:59 Target Range: Species: rat Total Number of Days: Total Number o f Hours: Total Number o f Data Points: T em perature 64'F to 79F 83 1962.0 1956 Relative Humidity 30% to 70% 83 1962.0 1956 Mean ( SD): Maximum: Median: Minimum: Number o f Points in Range (%): Number o f Points High (%): Number o f 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: DATE: 3-24f'Of 000923 ARGUS 418-018 :PAGE G-2 Tem perature D eviations Report Location: Room 17 Protocol N u m b er 418018 Range of Dates: 22-A ug-2000 14:44 to 12-Nov-2000 08:59 Tem perature Target Range: Species: rat Date 07-Sep-2000 07-Sep-2000 T im e 06:00 07:00 Temp. 63.8L 63.8L 64F to 79F Date Tim eTem p. H = Value out o f 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: 000924 ARGUS 418-018:PAGEG-3 Relative H um idity Deviations Report Location: Room 17 Protocol Number: 418018 Range o f Dates: 22-Aug-2000 14:44 to 12-Nov-2000 08:59 Hum idity Target Range: Species: rat Date 04-0ct-2000 Tim e 22:00 R.H. 70.6 H 30% to 70% Date Tim e H = V alue out o f range - High L = Value out o f range - Low R.H. = R elative Hum idity (%) R eport G enerated: 30-Nov-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: 000925 ARGUS 418-018:PAGE G-4 Tem perature and Relative Hum idity Report Location: Room 18 Protocol Num ber 418-018 Range o f Dates: 05-Sep-2000 14:00 to 03-Dec-2000 13:59 Target Range: Species: rat Total Number o f Days: Total Number o f Hours: Total Number o f Data Points: Tem perature 64F to 79F 90 2135.75 2129 Relative Hum idity 30% to 70% 90 2135.75 2129 Mean ( 1 SD): Maximum: Median: Minimum: Number o f Points in Range (%): Number o f Points High (%): Number o f 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: 5 ^ C S ( 000926 418-018:PAGE G-5 ARGUS 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 Tim e 05-0ct-2000 01:00 R.H. 70.2 H 30% to 70% Date Tim e R.H. H = Value out o f range - High L = Value out o f range - Low R.H. = Relative Hum idity (%) Report 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: 000927 APPENDIX H BIOANALYTICAL REPORTS (ANILYTICS) 000928 418-018:PAGE H -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: 12/08/2000 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 CHOLEST MG/DL 95 79 82 77 79 97 103 62 84.3 13.3 GLUCOSE MG/DL 117 116 89 87 99 90 94 99 98.9 11.7 LDL-DIR MG/DL 12 10 7 7 15 1 2 3 7.1 5 HDL-DIR MG/DL 44 61 65 51 54 13 18 40 43.3 19 TRIG MG/DL 322 151 116 176 156 771 559 257 313.5 233.6 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 + 000929 418-018:PAGE H-2 J f v l X J L 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: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Sgec. No. 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 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 + 000930 418-018:P A G E H -3 V jL J L - il. JL X \ A J 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 + 000931 418-018 .PAGE H-4 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 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 + 0Q0932 418-018.PAGE H-5 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 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 CHOLEST MG/DL 89 79 83 81 63 72 94 62 77.9 11.5 GLUCOSE MG/DL 95 109 99 145 100 109 84 110 106.4 17.9 LDL-DIR MG/DL 16 7 30 12 7 10 1 13 12 8.6 HDL-DIR MG/DL 61 61 37 69 49 34 12 50 46.6 18.4 TRIG MG/DL 111 95 196 48 140 294 632 91 200.9 190.1 Reference Range 48 - 74 - 0- 0- 3- 96 319 0 0 104 Page 5 + 000933 4 1 8 -018-.P A G E 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 PG/ML 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 000934 418-018:PAGE H-7 x 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: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT Accession Sgec. No. 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 + 00 .57 3.41 1.9 2.5 000935 418-018:PAGE H-8 XJL JL^CZ. J . X v A ^ 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 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 + 00 .57 3.41 1.9 2.5 000936 418-018:PAGE H-9 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: 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 T4,TOTAL T3.,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 000937 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 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 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- 7 100 0 .57 3.41 1.9 2.5 / Approved----------- -------- D a t e -- - Page 10 of 10 000938 418-018:PAGE H - l l j L . J I JL I 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: 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 + 000939 418-018:PA G E H -12 M 4 l JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client: AKGUS 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 ii i Ol 13 1 1 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 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 + 000940 418-018.P A G E H -13 4 t JL X W K / 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 - 74 - 0- 0- 3- 96 319 0 0 104 Page 3 + 000941 418-018:PA G E H -14 SJ, ^ j J i JL X w k 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: 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 + 000942 418-018:PAGE H-15 f v 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: 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 i i 1o 1u Sex Age CHOLEST GLUCOSE LDL-DIR HDL-DIR TRIG MG/DL MG/DL MG/DL MG/DL MG/DL 6F 6F 6F 6F 57 152 3 45 62 81 176 12 53 96 45 163 6 38 42 69 209 3 57 84 56.8 15.3 162.8 26.9 4.5 4.1 43.3 10.1 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 + 000343 418-018:PAGE H-16 ^Ji-LJl J. 200 Girard Street, Suite200, Gaithersburg, MD 20877 I NCORP ORATED 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 Sjsec. No. B 0030576 11004 B 0030577 11005 B 0030578 11006 B 0030579 11007 B 0030580 11008 Mean S.D. Group 6 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 + 000944 418-018:PAGE H-17 1i Ji i. J L W k / 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 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 T4,TOTAL T3,TOTAL FREE T3 UG/DL NG/DL PG/ML 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 1.927 .648 94.718 .908 8.185 .156 TSH NG/ML 1.18 3.92 1.26 0.00 1.66 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 + 00 .57 3.41 1.9 2.5 000945 418-018-.PAGEH-18 ^LAJL^CL 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: 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 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 + $00 418-018:PAGE H-19 ^ 4 A I. ^ l J . 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 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 000947 418-018:PAGE H-20 V L i - 1 --/ X JL J L \~ S K y 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORP ORATED 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 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 + 000948 418-018:PAGE H-21 2 i -Ji J . 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: 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 T3,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 0030569 10995 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 + 000949 418-018:PAGE H-22 ^ 1 v * JL i. w K 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: 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 37 - 51i00n0n- 00n - Approved------ -----------Page 12 of 12 -57 3.41 Date 21O..59K - -- 000950 418-018-.PAGEH-23 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: 12/14/2000 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 1F 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 48 96 Page 1 + 000951 418-018:PAGE H-24 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: 12/14/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 LID5 MILK Species: RAT Accession Spec. No. ID 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 + 000952 418-018.-PAGEH-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 + 000953 418-018:PAGE H-26 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: 12/14/2000 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 Gp Sex Age 6F 6F 6F CHOLEST HG/DL 0 6 7 4.3 3.8 B 0031059 10995 B 0031060 10997 Mean S.D. Group 7 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 + 000954 418-018:PAGE H-27 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 0031070 11008 Mean S.D. Group 8 Gp Sex Age 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 48 96 Approved -- ____ Page 5 of 5 D a t e --- cJ-L2q./ 000955 418-018:PAGE H-28 JL i . 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: 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 + 000956 418-018:PAGE H-29 1 JL I v A / 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 PUPS GD21 Species: RAT Accession Spec. No. ID 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 + 000957 418-018:PAGE H-30 \L L ~ A L A A 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: 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 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 96 74 319 Page 3 + 0 0 0- 30 125 000958 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 Sgec. No. 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 - 74 - 0- 0- 3- 96 319 0 0 125 Page 4 + 000959 418-018:PAGE H-32 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 Sgec. No. 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 - 74 - 0- 0- 3- 96 319 0 0 125 Page 5 + 000960 418-018:PAGE H-33 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 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 000961 418-018:PAGE H-34 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. i5 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 3.41 1.9 2.5 000362 418-018:PAGE H-35 ^3 JL 1 \ 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: 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 + 00 .57 3.41 1.9 2.5 000963 418-018:PAGE H-36 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 T4fTOTAL 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 + 0 0 .57 3.41 1.9 2.5 000964 418-018:PAGE H-37 <L / 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: 12/08/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT Accession Sgec. No. 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 3- 50 - 0- 7 100 0 .57 3.41 1.9 2.5 Approved -- Page 10 of 10 D a t e -- 000965 418-018:PAGE H-38 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 Spec. No. ID 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 1B8 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 + 000966 418-018:PAGE H-39 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 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 + 000967 418-018:PAGE H-40 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 Spec. No. ID 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 96 74 319 Page 3 + 0 0 0- 30 125 000968 418-018:PAGE H-41 i J i JL IvU 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 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 + 000969 418-018.PAGE H-42 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 Spec. NO. ID 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 + 000970 418-018:PAGE H-43 i L * l JL 1 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: 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 + 00 .57 3.41 1.9 2.5 000971 418-018:PAGE H-44 * J . 1VA/ 200 Girard Street, Suite 200, Gaithersburg, MO 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: 12/12/2000 Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LD5 Species: RAT Accession Spec. No. ID 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 000972 418-018:PAGE H-45 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 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 *37 - 50 inn Page 8 + 0f\ - .57 ^ A1 15.9* - 000973 418-018:PAGE H-46 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 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 + Q0974 418-018:PAGE H-47 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 7 100 0 .57 - 1.9 - 3.41 2.5 Approved -- -- Page 1 of 10 Dat. -- -/HJ- 000975 418-018:PAGE H-48 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: STUDY: REPORT TYPE: T 3 J Tt) fn, fr*), a f L AUDIT DATE: REPORT TO MANAGEMENT: AUDIT #: ) " 77'Oi Oh oL3 DATE SUBMITTED TO CLIENT ^ i. i /i i \ ri y ' it 900976 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 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 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F CHOLEST MG/DL 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 + 000977 418-018:PAGE H-50 c/ N SS 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 i1(9. 11 11 11 Accession Sgec. No. B 0035616 11628 B 0035617 11629 Mean S.D. Group 10 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 + Q00978 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 Sgec. No. 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 + 0Q0979 418-018:PAGE H-52 </ 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: 02/01/2001 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 + 0009S0 418-018-.PAGEH-53 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 0035565 11547 B 0035566 11552 Mean S.D. Group 4 O0 Sex Age CHOLEST MG/DL 4F 4F 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 + 0009S1 418-018.-PAGEH-54 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 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 Gp Sex Age 6F 6F 6F 6F 6F 6F 6F 6F CHOLEST MG/DL 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 + 000982 418-018:PAGE H-55 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 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 y Approved----- .T-------- Page 7 ''of 7 Date -- 000983 418-018:PAGE H-56 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: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 DAMS LD5 Species : RAT Accession Spec. No. i6 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 1 + 000984 418-018 :PAGE H-57 SJJ-*JL JL x 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: 02/02/2001 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 P 11 P 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 P 12 P 12 P 12 P 12 F 12 P 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 000985 418-018:PAGE H-58 M i LJl 1 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: 02/02/2001 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 000986 418-018:PAGE H-59 1 a -Jl J . 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: 02/02/2001 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 T4,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 + 418-018:PAGE H-60 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 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 2.5 418-018:PAGE H-61 ^4 a JL X w W 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: 02/02/2001 Date Reported: 02/13/2001 Client No. 1028 Study: 418018 DAMS LD5 Species: RAT Accession Sjaec. 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 Page 6 of 6 mtr P009S9 418-018:PAGE H-62 ^4L*dl A A 200 Girard Street, Suite200, Gaithersburg, MD 20877 I NCORP ORATED 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 + 000990 418-018.PAGE H-63 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 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 000991 418-018:PAGE H-64 J . .X V --A * J 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 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 + 000992 418-018:PAGE H-65 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 B 00003355889976 1111558821 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 090993 418-018:PAGE H-66 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 Sgec. No. 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 Gp Sex Age 7P 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 B 0035915 11600 B 0035916 11601 B 0035917 11602 B 0035918 11603 B 0035919 11604 B 0035920 11605 B 0035921 11606 Reference Range 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 + 000994 418-018:PAGE H-67 Jm JLL X J L v A 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 DAMS LD5 Species: RAT Accession Sgec. No. 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 S 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 000395 418-018 .PAGE H-68 \JJLk~JLL 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 Spec. No. ID 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 l 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 + 00099b 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 Sgec. No. Gp Sex Age B 0035742 11648/651 12 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 + 00 20 80 3125 U0937 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, IHC. 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 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 + 3S8 418-018 :PAGE H -71 cA N fi INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. Date Collected: 905 SHEEHY DRIVE Date Received: 02/ 02/2001 BUILDING A HORSHAM. PA 19044(215) 443-8710 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 Approved Page' 4 of Date 418-018:PAGE H-73 M j / J l 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 PUPS LD5 Species: RAT Accession Sgec. No. 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 + ih u o o o 418-018:PAGE H-74 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: AKGUS 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 OiOOl 418-018:PAGE H-75 ^ J ' v f JL J L V A 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 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 74 319 00 Approved -- Page 3 of 3 20 80 Date 00X002 418-018 .PAGE H-76 I j j _ 2 v A J 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: AR6I/& STODY: REPORT TYPE: o/if/n, n, % rftzrz men AUDIT DATE: REPORT TO MANAGEMENT: AUDIT #: &'}?-0) 0)~0i$ \ 418-018:PAGEH- I vf X Iv A J 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: CH0L STUDY: (Jfiotf AUDIT DATE: REPORT TO MANAGEMENT: o) AUDIT #: 001004 418-018:PAGE H-78 S J^U L*dJL JL I V v 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: 02/07/2001 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 l + 0 0 00 001005 418-0i8: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 Sgec. No. 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 OCIOOS 418-018:PAGE H-80 X 200 Girard Street, Suite 200, Gaithersburg, MO 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: 02/07/2001 Date Reported: 10/10/2001 Client No., 1028 Study: 418018 F0 C/S GD21 Species: RAT Accession Sgec. No. 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- 0- 0- 00000 Page 3 + 001007 418-018:PAGE H-81 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 H(O2R1S5)HAM4,43-P8A71019044- 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 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 00 00 Page 4 + 0 0 0 0 OCIOOS 418-018:PAGE H-82 \J*JL^LL J . i . w k J INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 F a x :3 0 1 -9 7 7 -0 4 3 3 C lie n t: ARGUS RESEARCH LABORATORIES, 9 05 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . Date C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 1 0 /1 0 /2 0 0 1 C lie n t No. 1028 S tu d y : 4 1 8 0 1 8 F0 C /S GD21 S p e c ie s : RAT A ccession Spec. No. i5 B 0037203 B 0037204 B 0037205 B 0037206 B 0037207 B 0037208 B 0037209 B 0037210 10985 10987 10988 10989 10990 10991 10992 10993 Mean S.D. Group 7 Gp S ex Age 7F 7F 7F 7F 7F 7F 7F 7F L D L -D IR MG/GM 0.46 0.78 0.81 1.03 0.65 0.75 0.92 0.92 .79 .1 7 8 H D L -D IR MG/GM 0.45 0.59 0.63 0.69 0.51 0.57 0.69 0.69 .603 .0 9 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 R eferen ce Range 0- oooo o 00 Approved -- Page 5 o f 5 D a te 001009 418-018:PAGE H-83 C lie n t: " JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 3 0 1 -9 2 1 -0 1 6 8 F a x :3 0 1 -9 7 7 -0 4 3 3 ARGUS RESEARCH LABORATORIES, 9 0 5 SHEEHY DRIVE BUILDING A HORSHAM. PA 1 9 0 4 4 (215) 443-8710 INC . D ate C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 Date R eported: 1 0 /3 0 /2 0 0 1 C lie n t No. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP1 S p e c ie s :: RAT A ccession 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 S ex Age 1F 1F 1F 1F 1F 1F LD L-D IR MG/GM 0.00 0.00 0.06 0.00 0.00 0.00 .0 1 .024 H D L -D IR 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 .1 2 TRIG MG/GM 9.43 12.82 10.39 14.78 8.18 16.39 1 1 .9 9 8 3.208 B 0036813 B 0036814 B 0036815 B 0036816 B 0036817 B 0036818 B 0036819 B 0036820 B 0036839 11019 11021 11023 11024 11025 11026 11027 11028 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 1 1 .0 5 9.13 1 0 .1 2 7.56 9.96 9.02 1.249 R e fe ren c e Range 0- 000 Page 1 + 00 00 OOlOiO 418-018:PAGE H-84 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 C lie n t: ARGUS RESEARCH LABORATORIES, 905 SHEEHY D R IV E BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 1 0 /3 0/2 0 01 C lie n t No. 1028 S tu d y : 4 18 0 18 F0 LD5 REP1 S p e c ie s : RAT A ccession Sgec. No. B 0036821 B 0036822 B 0036823 B 0036824 B 0036825 B 0036826 B 0036827 B 0036828 B 0036840 11029 11031 11032 11033 11034 11036 11037 11038 11035 Mean S.D. Group 11 Gp S ex Age 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F 11 F L D L -D IR MG/GM 0.15 0.31 0.33 0.01 0.00 0.00 0.00 0.08 0.10 .1 0 9 .131 H D L -D IR 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 T R IG 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 R e fe re n c e Range 13 F 0.19 00 0.45 00 3.26 00 9.58 00 Page 2 + (JC lO ll 418-018:PAGE H-85 C lie n t: INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921 -0168 Fax: 301 -977-0433 ARGUS RESEARCH LABORATORIES, 905 SHEEHY D R IV E BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 1 0 /3 0 /2 0 0 1 C l i e n t No., 1028 S tu d y : 418 0 18 F0 LD5 REPI S p e c ie s : RAT A ccession No. B 0036834 11055 B 0036835 11057 B 0036836 11061 B 0036837 11064 B 0036838 11065 Mean S.D. G roup 13 l 0 OoTnH 1 Gp S ex Age 13 F 13 F 13 F 13 F 13 F L D L -D IR MG/GM 0.00 0.00 0.00 0.13 0.00 .053 .085 H D L -D IR 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 T R IG MG/GM 22.98 15.80 19.26 11.96 15.61 15.865 4.837 B 0036763 B 0036764 B 0036765 B 0036766 B 0036767 10921 10922 10923 10924 10927 Mean S.D. Group 2 2F 2F 2F 2F 2F 0.11 0.00 0.14 0.00 0.00 .0 5 .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 1 1 .7 3 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 .1 1 6 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 R e fe re n c e Range 0- 000 Page 3 + 0 0 00 ^01012 418-018:PAGE H-86 C lie n t: omcs INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MO 20877 301 -921 -0168 Fax: 301 -977-0433 ARGUS RESEARCH LABORATORIES, 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 1 0 /3 0 /2 0 0 1 C l i e n t No.. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REPI S p e c ie s : RAT A ccession Sgec. No. B 0036772 B 0036773 B 0036774 B 0036775 B 0036776 B 0036777 10943 10944 10948 10952 10955 10956 Mean S.D. Group 4 Gp Sex Age 4F 4F 4F 4F 4F 4F L D L -D IR MG/GM 0.00 0.00 0.00 0.13 0.00 0.00 .022 .053 HDL--D IR MG/GM 0.26 0.37 0.42 0.48 0.23 0.31 .345 .096 CHOLEST MG/GM 2.95 6.56 3.51 5.52 2.89 3.27 4.117 1.542 TRIG MG/GM 18.57 14.80 1 5 .1 5 13.06 9.21 10.87 13.61 3.333 B 0036778 B 0036779 B 0036780 B 0036781 B 0036782 B 0036783 10957 10966 10967 10968 10969 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 9.60 7.95 10.65 7.51 8.80 9.03 8.957 1 .1 5 9 B 0036784 10971 B 0036785 10973 R eferen ce 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 + 00101J 418-018:PAGE H-87 C lie n t; INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, M D 20877 3 0 1 -9 2 1 -0 1 6 8 Fax: 301-977-0433 ARGUS RESEARCH LABORATORIES, 905 SHEEHY D R IV E BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . Date C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 1 0 /3 0/2 0 01 C lie n t No. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP1 S p e c ie s : RAT A ccession Sgec. NO. B 0036786 10977 B 0036787 10983 B 0036788 10979 B 0036789 10984 Mean S.D. Group 6 Gp S ex Age 6F 6F 6F 6F LDL-D IR MG/GM 0.00 0.01 0.16 0.00 .028 .065 H D L -D IR 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 1 1 .7 6 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 B 0036796 B 0036797 B 0036798 B 0036799 10999 11000 11001 11002 11003 R e fe re n c e 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 + 0C1014 418-018:PAGE H-88 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 05 SHEEHY DRIVE BUILDING A HORSHAM. PA 1 9 0 4 4 (215) 443-8710 IN C . Date C o lle c te d : Date R eceived: 0 2 /0 7 /2 0 0 1 Date R eported: 1 0 /3 0 /2 0 0 1 C lie n t No. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 R E P I S p e c ie s : RAT A ccession Sgec. No. B 0036800 11004 B 0036801 11005 B 0036802 11006 B 0036803 11007 B 0036804 11008 Mean S.D. Group 8 Gp Sex Age 8P 8F 8F 8F 8F L D L -D IR MG/GM 0.00 0.00 0.00 0.00 0.15 .015 .047 H D L -D IR 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 .1 5 5 T R IG MG/GM 10.48 11.49 7.95 7.79 8.06 8.986 1.386 B 0036805 B 0036806 B 0036807 B 0036808 B 0036809 B 0036810 B 0036811 B 0036812 11009 11010 11011 11012 11013 11015 11016 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 R eferen ce Range 00 Approved -- Page 0- 0 0 yQ u z z i____ of 6 D a te -JS/SS& l 418-018.PAGE H-89 -f # -Ji i . x V . / L / INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 F a x :3 0 1 -9 7 7 -0 4 3 3 C lie n t: ARGUS RESEARCH LABORATORIES, 9 0 5 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2/0 7 /2 0 0 1 D ate R eported: 10/1 0/2 0 01 C l i e n t No.. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. B 0036600 B 0036601 B 0036602 B 0036603 B 0036604 B 0036605 B 0036606 B 0036607 B 0036608 B 0036609 B 0036610 B 0036611 B 0036612 B 0036613 11501 11502 11503 11504 11505 11506 11507 11508 11509 11510 11511 11512 11513 11514 Mean S.D. Group 1 Gp Sex Age 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F 1F L D L -D IR 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 H D L -D IR 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 .7 1 6 .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 B 0036713 B 0036714 B 0036715 B 0036716 B 0036717 B 0036718 11620 11621 11622 11623 11624 11625 11626 R eference 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 00 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 + 001016 418-018:PAGE H-90 Mi L J l x I v U INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 05 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 1 0 /1 0/2 0 01 C lie n t No. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession 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 L D L -D IR MG/GM 0.00 0.00 0.00 0 0 H D L -D IR MG/GM 0.16 0.15 0.15 .133 .027 CHOLEST MG/GM 3.30 1.83 1.86 2 .1 1 6 .459 TRIG MG/GM 59.25 7.40 10.48 14.211 16.528 B 0036722 B 0036723 B 0036724 B 0036725 B 0036726 B 0036727 B 0036728 B 0036729 B 0036730 11630 11632 11633 11634 11635 11636 11637 11638 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 R eference 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 + 001017 418-018:PAGE H-91 %-L J - J A . X x v A ^ INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MO 20877 301-921-0168 Fax:301-977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 0 5 SHEEHY D R IVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 1 0 /1 0 /2 0 0 1 C l i e n t No., 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. B 0036733 B 0036734 B 0036735 B 0036736 B 0036737 B 0036738 B 0036739 B 0036740 B 0036741 B 0036742 B 0036743 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 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 L D L -D IR 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 H D L -D IR 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 B 0036745 B 0036746 B 0036747 B 0036748 B 0036749 B 0036750 B 0036751 B 0036752 B 0036753 11654 11655 11656 11657 11658 11659 11660 11661 11662 11663 R eferen ce 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 + oc1018 418-018:PAGE H-92 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 05 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . Date C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R ep o rted : 10/ 10/2001 C l i e n t No., 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. B 0036754 11664 B 0036755 11665 B 0036756 11667 Mean S.D. G roup 13 Gp Sex Age 13 F 13 F 13 F L D L -D IR MG/GM 0.00 0.08 0.00 .015 .03 H D L -D IR 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 B 0036615 B 0036616 B 0036617 B 0036618 B 0036619 B 0036620 B 0036621 B 0036622 B 0036623 B 0036624 B 0036625 B 0036626 11515 11516 11517 11518 11519 11520 11521 11523 11524 11525 11526 11527 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 R eference Range 0- 000 Page 4 + 00 00 001013 418-018:PAGE H-93 J i v l JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 0 5 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . Date C o lle c te d : D ate R eceived: 0 2/0 7 /2 0 0 1 Date R eported: 1 0 /1 0/2 0 01 C l i e n t No.. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. B 0036627 B 0036628 B 0036629 B 0036630 B 0036631 B 0036632 B 0036633 B 0036634 B 0036635 B 0036636 B 0036637 B 0036638 B 0036639 B 0036640 11529 11530 11532 11533 11534 11535 11536 11537 11538 11539 11540 11541 11542 11543 Mean S.D. Group 3 Gp Sex Age 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F L D L -D IR 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 H D L -D IR 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 B 0036641 B 0036642 B 0036643 B 0036644 B 0036645 B 0036646 B 0036647 11544 11545 11546 11547 11549 11550 11551 R eferen ce Range 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 + 001020 418-018:PAGE H-94 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 05 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2 /0 7 /2 0 0 1 D ate R eported: 10/1 0/2 0 01 C l i e n t No.. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. B 0036648 B 0036649 B 0036650 B 0036651 B 0036652 B 0036653 11552 11553 11554 11555 11556 11557 Mean S.D. Group 4 Gp Sex Age 4F 4F 4F 4F 4F 4F L D L -D IR MG/GM 0.03 0.00 0.00 0.00 0.00 0.00 .006 .013 H D L -D IR MG/GM 0.14 0.1 1 0.06 0.12 0.12 0.08 .1 0 5 .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 1 1 .8 3 15.948 6.227 B 0036654 B 0036655 B 0036656 B 0036657 B 0036658 B 0036659 B 0036660 B 0036661 B 0036662 B 0036663 B 0036664 11558 11559 11561 11563 11564 11565 11567 11568 11569 11570 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 R eference Range 0- 000 Page 6 + 0 0 0 0 OVlOi.1 418-018:PAGE H-95 ^ rf * JL X V A ^ INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MO 20877 301-921-0168 Fax:301-977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 05 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2/0 7 /2 0 0 1 Date R eported: 1 0/1 0 /2 0 0 1 C l i e n t No,. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. B 0036665 B 0036666 B 0036667 B 0036668 B 0036669 B 0036670 B 0036671 B 0036672 B 0036673 B 0036674 B 0036675 B 0036676 B 0036677 B 0036678 11572 11573 11574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 Mean S.D. Group 6 Gp S ex Age 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F 6F L D L -D IR 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 H D L -D IR 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 B 0036680 B 0036681 B 0036682 B 0036683 B 0036684 B 0036685 11586 11587 11588 11589 11590 11591 11592 R eference 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 1 8 .1 7 7.11 7.80 13.17 00 Page 7 + 001022 418-018:PAGE H-96 > 4 4 L * J l JL i V V j INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 05 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . D ate C o lle c te d : D ate R eceived: 0 2/0 7 /2 0 0 1 Date R eported: 10/1 0/2 0 01 C lie n t No. 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. B 0036686 B 0036687 B 0036688 B 0036689 B 0036690 B 0036691 B 0036692 11593 11594 11595 11596 11597 11598 11599 Mean S.D. Group 7 Gp Sex Age 7F 7F 7F 7F 7F 7F 7F L D L -D IR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 H D L -D IR MG/GM 0.14 0.16 0.14 0.14 0.19 0.22 0.16 .153 .033 CHOLEST 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 B 0036694 B 0036695 B 0036696 B 0036697 B 0036698 B 0036699 B 0036700 B 0036701 B 0036702 11600 11601 11602 11603 11604 11605 11606 11607 11608 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 .1 1 5 .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 1 0 .1 5 7.80 9.19 5.41 8.55 7.826 1.846 R eference Range 0- 000 Page 8 + 00 00 001023 418-018:PAGE H-97 I cyiW dm cs INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921-0168 Fax: 301 -977-0433 C lie n t: ARGUS RESEARCH LABORATORIES, 9 0 5 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 (215) 443-8710 IN C . Date C o lle c te d : Date R eceived: 0 2/0 7 /2 0 0 1 Date R eported: 1 0/10/2001 C l i e n t No., 1028 S tu d y : 4 1 8 0 1 8 F0 LD5 REP2 S p e c ie s : RAT A ccession Sgec. No. 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 L D L -D IR MG/GM 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 0 H D L -D IR 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 .9 1 1 .243 TRIG MG/GM 1 1 .9 8 6.11 16.42 7.18 8.63 6.32 9.22 9.19 7.77 9.202 3.243 R eference Range 0- o- 0- o00oo -iSM&MUA pproved _____ ______________ ----- D a te Page 9 o f 001024 418-018:PAGE H-95 ^ v * JL J L 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 + 001025 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 Sgec. No. 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 CHOLEST 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 00 00 Page 8 + 0 0 0 0 001026 418-018.PAGE H-97 % L 1 1 \A ^ 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 1 9 0 4 4 - (215) 443-8710 Date Collected: Date Received: 0 2 / 0 7 / 2 0 0 1 Date Reported: 1 0 / 1 0 / 2 0 0 1 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 TRIG MG/GM MG/GM 2.06 1.73 2.36 1.65 1.97 1.76 1.73 2.17 1.77 11.98 6.11 16.42 7.18 8.63 6.32 9.22 9.19 7.77 1.911 .243 9.202 3.243 Reference Range 00 0 0 0- 0 0- 0 Approved -- Page 9 of 9 Date SjJiilLL 001027 4 1 8 -018.-P A G E 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 FI C/S GD21 Species: RAT Accession Spec. No. ID 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 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 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 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 0.02 0.02 0.01 0.03 0.03 0.02 0.02 0.02 .021 .006 3.28 3.21 3.70 3.18 4.32 3.26 3.32 3.36 3.454 .385 5.06 5.13 6.04 5.95 7.90 5.18 6.16 5.48 5.863 .931 Reference Range 00 00 Page l + 0 0 00 001028 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 Fi C/S GD21 Species: RAT Accession Sgec. No. 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 0- 0- 0- 00000 Page 2 + 001029 418-018:PAGE H -100 o VZH qHYTICS 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 FI C/S GD21 Species: RAT Accession Sjoec. No. 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 00 Page 3 + 0 0 00 001030 418-018:PAGE H-101 " v A. X 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) 4431-8710 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. I 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 Reference Range 0- 0- 0- 00 00 0 Page 4 + 001031 418-018-.PAGE H -102 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 00 00 00 Approved -----Page 5 o / Date UC1032 418-018 .-P A G E H -1 0 3 MmJLL JL 1 V V J 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCOR PORA 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: 02/07/2001 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 + q0 \ 0 5 3 418-018:PAGE H-104 Z f. J, X v A J 200 Girard Street, Suite 200, Gaithersburg, MD 20877 I NCORP ORATE 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: 02/07/2001 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 00 00 Page 2 + o0 00 001034 418-018.PAG E 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: 0 2 / 0 7 / 2 0 0 1 Date Reported: 10/ 10/2001 Client No. 1028 Study: 418018 Fl LD5 REP1 Species: RAT Accession Sgec. No. 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 10.3K 3.964 Reference Range 0- 0- 0- 00000 Page 3 + 00X035 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 905 SHEEHY DRIVE BUILDING A HORSHAM, PA 19044(215) 443-8710 INC. 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 o0 Page 4 + 0 0 00 001036 418-018:PAGE H-107 </ m INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIESf 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 REP1 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 CHQLEST 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 0- 0- 0- 00000 Page 5 + 001037 418-018:PAGE H -108 c/ m INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301 -921 -0168 Fax: 301 -977-0433 Client: ARGUS RESEARCH LABORATORIES, IHC. 9Q5 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 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 + 001038 418-018:PAGE H-109 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-068 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: 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REP1 Species: RAT Accession Sgec. No. 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 + OGlOoS 418-018:PAGE H-110 c /I 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: 02/07/2001 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 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 7M 7M 8F 8F 8F 8F 8F 8F 8F 8F 8F 8F 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 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 + 001.040 418-018:PAGE H -l 11 JL X 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: 10/10/2001 Client No. 1028 Study: 418018 Fl LD5 REPI Species: RAT Accession Sgec. No. B 0037054 10999 B 0037055 11000 B 0037056 11001 B 0037057 110Q2 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 0- 0 0- 0 Page 9 + 0 0 00 001041 418-018-.PAGE H-112 zJM S 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 REP1 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 0- 0- 0- 00000 Approved ----------,)-d-------------- nate -J2jjsJ*L Page 1-0 of 10 001042 418-018:PAGE H-113 JL 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: 02/08/2001 Date Reported: 10/10/2001 Client No,, 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Sgec. No. 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 1P 1P 1F 1P 1F 1F 1F 1F 1F 1F 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 + 001043 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 FI LD5 REP2 Species: RAT Accession Sgec. No. 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- 000 Page 2 + 0 0 00 001044 418-018:PAGE H-115 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 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 0- 0- 0- 00000 Page 3 + 001045 418-01&PAGE H-l 16 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 3 0 1 -9 2 1 -0 1 6 8 F a x :3 0 1 -9 7 7 -0 4 3 3 Client: ARGUS RESEARCH LABORATORIES, INC. 9 0 5 SHEEHY DRIVE BUILDING A HORSHAM, PA 1 9 0 4 4 - (2 1 5 ) 4 4 3 -8 7 1 0 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 Sjsec. 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 + 001046 418-018:PAGE H-117 V -L JL~dL -L x V A - / INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 3 0 1 -9 2 1 -0 1 6 8 F a x :3 0 1 -9 7 7 -0 4 3 3 Client: ARGOS 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 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 + 001047 418-018:PAGE H-118 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 3 0 1 -9 2 1 -0 1 6 8 Fax: 301-977-0433 Client: ARGUS RESEARCH LABORATORIES, INC. 905 SHEEHY DRIVE BUILDING A HORSHAM. FA 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 0- 0- 0- 00000 Page 6 + 001048 418-018:PAGE H-119 c/ m INCORPORATED 200 Girard Street, Suite 2 00 , Gaithersburg, MD 20877 3 0 1 -9 2 1 -0 1 6 8 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 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 0- 000 Page 7 + 0 0 0 0 001049 418-018:PAGE H-120 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: 02/08/2001 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 FI LD5 REP2 Species: RAT Accession Sgec. No. 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 0- 0- 0- 00000 Page 8 + 001050 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 Fl 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 19.35 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- 0 Page 9 + 0- 0 0- 0 001051 418-018:PAGE H-122 " JL JL INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 Fax:301-977-0433 Client; ARGUS RESEARCH LABORATORIES, IRC, 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 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 CHDLEST 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 001052 418-018:PAGE H-123 v*L J--JJL JL J L 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: 02/08/2001 Date Reported: 10/10/2001 Client No., 1028 Study: 418018 Fl LD5 REP2 Species: RAT Accession Sgec. No. 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 Gp Sex Age 8F 8F 8F 88 F F 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 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 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 00..0190 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- 000 Page 11 + 00 00 001053 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 B 0037399 0037400 1111661189 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 00 Approved -- Page 0- 0 0 /0 ___ 12 of 12 00 Date --i&pApJL 001054 418-018:PAGE H-125 11 V I i f INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0188 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 m. STUDY: HHOit AUDIT DATE REPORT TO MANAGEMENT A U D IT # 0\^QL 001055 APPENDIX I BIOANALYTICAL REPORTS (PRIMEDICA WORCESTER, SPONSOR AND SRI) 001056 418-018-.PAGEI-1 FINAL REPORT VALIDATION OF A GAS CHROM ATOGRAPHIC-M ASS SPECTROM ETRIC M ETHOD FO R THE ANALYSIS OF M EV A LO N IC ACID IN EDTA R A T PLASMA FO R PRIM ED ICA ARGUS STUDY 418-018 Primedica-Worcester Project Number: PABX-BVA PRIMEDICA ARGUS 905 Sheehy D rive, Building A Horsham, PA 19044 00105? 418-018:PAGE 1-2 O P rim edica FINAL REPORT VALIDATION OF A GAS CHROM ATOGRAPHIC-M ASS SPECTRO M ETRIC M ETHOD FOR THE ANALYSIS O F M EVALONIC ACID IN EDTA RAT PLASMA FO R PRIM EDICA ARGUS STUDY 418-018 Primedica-Worcester Project Number: PABX-BVA Submitted to: PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 Submitted by: Prim edica-W orcester 57 Union Street Worcester, MA 01608 Primedica-Worcester Report # PABX-BVA-01-62 Page 1 o f 184 Issue Date: November 7,2001 Mark A. Netsch / Date Project Scientist Department o f Analytical Chemistry Primedica-Worcester 1)01058 418-018:PAGE 1-3 C/Primedica 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 o f good laboratory practice. Official Journal o f 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 o f the project or the interpretation o f the results in this report. 5 ^/ Mark A. Netsch / Date Project Scientist f'/y/ar 001059 418-018:PAGE 1-4 GPrim ekca Primedica-Worcester Project number. PABX-BVA Page 3 Final Report_________________________ ________ For Primedica Argus Study Num ber 418-018 QUALITY ASSURANCE STATEMENT The following are the inspection dates and report dates o f QAU audit/inspections for Validation O f A Gas Chromatographic-Mass Spectrometric Method For The Analysis O f Mevalonic Acid In EDTA Rat Plasma For Primedica Argus Study 418-018, Project Number PABX-BVA. Critical Phases 1. Laboratory Procedures D ate Inspected 10/19/2000 Date Report Subm itted to Project Scientist M anagement 10/20/2000 10/23/2000 1. Raw Data 11/07/2000 04/06/2001 04/06/2001 2. Draft Final Report 03/26/2001 03/26/2001 04/04/2001 The Final Report for Validation o f A Gas Chromatographic-Mass Spectrometric Method For The Analysis O f M evalonic 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 o f good laboratory practice. Official Journal o f European Communities: Legislation 32 (No. L 315; 28 October): 1-17, on 11/7/01. The results as presented accurately reflect the raw data. ----- ------------------------ L___ Quality Assurance Auditor Date U010GG 418-018:PAGE 1-5 OPrim edta Primedica-Worcester Project number: PABX-BVA Page 4 Final Report_________________________________ For Primedica Argus Study Number: 418-018 TABLE OF CONTENTS Pape N o. COMPLIANCE STATEMENT.................................................................................................. 2 QUALITY ASSURANCE STATEMENT.............................................................................. 3 LIST OF TABLES AND APPENDICES.................................................................................. 5 CONTRIBUTING PERSONNEL.............................................................................................. 7 ANALYTICAL REFERENCE STANDARD CHARACTERIZATION/STABILITY.....8 ARCHIVAL STORAGE..................... 9 ABSTRACT................................................................................................................................. 10 INTRODUCTION.......................................................................................................................11 EXPERIMENTAL DESIG N .....................................................................................................11 O verview ...................................................................................................................................11 Materials and M ethods............................................................................................................11 General Comments................................................................................................................ 11 Validation D esign.................................................................................................................... 11 Validation D escription........................................................................................................ 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 CC'lOol S 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 o f Interpolated Mevalonic Acid Lactone QC Standard Concentrations with Inter and Intra-Day Summary Statistics............................................................. 22 TABLE 2 Summary o f Back-Calculated M evalonic Acid Lactone Calibration Standard Concentrations............................................................................................................... 24 TABLE 3 M evalonic Acid Lactone Extraction Data & Recovery Calculations..................... 25 TABLE 4 Internal Standard Extraction Data & Recovery Calculations.................................. 26 TABLE 5 Summary o f Least-Squares Linear Regression Constants and Analysis D ates.....27 TABLE 6 Summary o f QC Standard 72-Hour Room Temperature Stability in Matrix.........28 TABLE 7 Summary o f QC Standard Freeze/Thaw (3 Cycles) Stability in Matrix.................29 TABLE 8 Summary o f Interpolated M evalonic Acid Lactone Dilution QC Concentrations for Dilution Validation...................................................................................................30 001062 418-018:PAGE 1-7 C/Prim eota Primedica-Worcester Project number PABX-BVA Page 6 Final Report__________________________________For Primedica Argus Study Number 418-018 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 o f Control Blank EDTA Rat Plasma Concentrations.............................33 TABLE 12 Summary o f Unique Lots o f EDTA Plasma Concentrations................................... 34 TABLE 13 Summary o f Internal Standard Peak Areas.................................................................35 TABLE 14 Summary o f 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 001063 418-018:PAGE 1-8 OPrjmedica Primedica-Worcester Project number PABX-BVA Page 7 Final Report_________________________________ For Primedica Argus Study Number 418-018 CONTRIBUTING PERSONNEL Project Scientist........................ Research A ssociate................... Director, Analytical Chemistry Report Coordinator................... . Marie A. Netsch, B.A. .....Heidi Gagnon, B.S. James A. Jersey, Ph.D. ........Brenda L. Brooks 001064 418-018:PAGE 1-9 ^ P rimedica Primedica-Worcester Project num ber'PABX-BVA Page 8 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. D L -M evalonolacetone-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 o f the analytical standard and internal standard is the responsibility o f the Supplier, as is the method o f synthesis, fabrication or derivation and stability determination. 01065 418-018:PAGE 1-10 GPrim edta Primcdica-Worcestcr 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 w ill be maintained for a minimum period o f five years following submission o f the final report in the Primedica-Worcester Archives in Worcester, MA. After five years the Sponsor w ill be contacted for disposition instructions. Archival material w ill be indexed by Primedica-Worcester Report #PABX-BVA-01-62. 001066 418-018-.PAGE 1-11 OPrim edta 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 o f mevalonic acid in EDTA rat plasma. The procedure involves the conversion o f mevalonic acid (MVA) to mevalonic acid lactone (MVL) and the analysis o f a liquid/Iiquid extract by gas chromatography coupled with mass spectrometry (GC/MS) using positive chemical ionization (PCI). A mathematical factor to convert sample concentrations o f 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-tLj, was used for quantification. The results for the validation indicate that the method is sufficiently linear, specific, reproducible and accurate to support the analysis o f mevalonic acid in EDTA rat plasma samples. 001067 418-018:PAGE 1-12 OPrimedica Primedica-Worcester Project number PABX-BVA Page 11 Final Report_____________ ____________________ For Primedica Argus Study Number 418-018 IN T R O D U C T IO N : The objective o f this study was to develop and validate an analytical method for determining levels o f mevalonic acid (MVA) in EDTA rat plasma. EXPERIM ENTAL DESIGN: O verview : The assay described in this report employed the conversion o f m evalonic acid (MVA) to mevalonic acid lactone (MVL) and the analysis o f liquid/liquid EDTA rat plasma samples by GC/MS using positive chemical ionization. The internal standard, MVLkU, was used for quantitation. The validated assay covered the concentration range o f 10.0 ng/mL to 250 ng/mL o f MVL in EDTA rat plasma using 100 pL sample volumes. M aterials and M ethods: Refer to Appendix A for a detailed description o f materials and methods employed during the course o f this validation study. In addition, quality control (QC) standards were prepared in water at the lower limit o f quantiation (LLOQ QC, 10.0 mg/mL) using the same stock solutions used for the water QC standards. A set o f 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 o f 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. V alidation Design: V alidation Description: The assay was comprised o f five analytical runs. Three o f the five analytical runs were comprised o f bracketing seven-point water matrix calibration curves with six replicates o f water matrix quality control (QC) samples at four concentrations, six replicates o f EDTA rat plasma matrix QC samples at three concentrations and six replicates o f 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 0010G8 418-018:PAGE 1-13 O P rlmedica Primedica-Worcester Project number PABX-BVA Page 12 Final Report_________________________________ For Primedica Argus Study Number: 418-018 run. An analysis o f the method recovery was included in the third analytical run, which consisted o f an extracted calibration curve and a solvent calibration curve. The analysis o f freeze/thaw (F/T) stability and room temperature (RT) stability, both in water and matrix, was included in the fourth analytical run. The filth analytical run was a re-injection o f 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 Rim 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 Param eters 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 o f quantitation (LLOQ) o f 10 ng/mL. This level o f background concentration would significantly skew the results o f plasma QC's at the lower concentration levels. Therefore, calibration and QC standards were prepared in water matrix to evaluate precision, accuracy and linearity o f the method. The second set o f QC's was prepared at three concentration levels in EDTA rat plasma to evaluate the ability o f the method to extract and quanitate MVL equally from water or plasma matrices. The plasma QC's were also used to evaluate matrix stability. 001069 418-018:PAGE 1-14 OPrim edta 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 o f six replicates o f each QC standard concentration levels in both water and plasma. Mean peak area counts o f 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 lim its 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 o f 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 o f High QC standards in both water and plasma. The endogenous concentration o f mevalonic acid was low compared to the high QC concentration and did not make a significant contribution to the concentration results. Mean concentrations o f m evalonic 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 o f the method were evaluated at four concentrations by analyzing six replicates o f the QC standards prepared in water over the course o f 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 o f a QC standard diluted 1:5 water. Intra-day precision and accuracy was evaluated and reported as % RSD and % bias, respectively. Recovery o f 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 assinning 100% recovery. Linearity o f the method was evaluated by visual inspection o f the calibration curve and examination o f 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 o f quality control samples prepared at two levels, which were subjected to three F/T cycles, prior to their extraction and analysis. o o i o -. o 418-018:PAGE 1-15 GPrimedtca Primedica-Worcestcr 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 o f 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 o f final extracts was evaluated by re-analyzing the day-one validation extracts consisting o f duplicate calibration curves, six replicates o f 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/Plasma Extraction Equivalency: The ability o f the method to extract and quantitate mevalonic acid equivalently from water or plasma matrices was evaluated. The equivalence was evaluated by the analysis o f six replicates o f each o f three QC standard levels in water and plasma during the first three validation runs. Mean peak area counts o f 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 o f results. The comparison was also evaluated by the analysis o f six replicates o f high-QC standards in water and plasma during the first three validation runs. A t this concentration level (200 ng/mL) the endogenous level o f mevalonic acid was not a significant interference. Mean concentrations o f 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 o f results. The percent difference results were all within the acceptable limits o f 10%. The extraction o f mevalonic acid lactone from water and plasma are equivalent. Therefore, U01071 418-018:PAGEI-16 GPrimedica 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 m evalonic acid lactone concentration levels in plasma. Accuracy: Inter-day and intra-day accuracy were acceptable for the assay o f Mevalonic acid lactone in water over the concentration range o f 10.0 ng/mL to 250 ng/mL. Intra-day accuracy was evaluated by the analysis o f six replicates o f each o f 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 o f 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 o f results. Inter-day accuracy was evaluated by the analysis o f six replicates o f each o f 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 o f 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 o f results. P recisio n : Inter-day and intra-day precision was acceptable for the assay o f mevalonic acid lactone in water over the concentration range o f 10.0 ng/mL to 250 ng/mL. Intra-day precision was evaluated by the analysis o f six replicates o f each o f four QC standard levels in water during the first three validation runs. The intra-assay variance estimate o f 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 o f results. Inter-day precision was evaluated by the analysis o f six replicates o f each o f four QC standard levels in water during the first three validation runs. The inter-assay variance estimate o f 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 o f results. 001072 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 o f results. Dilution QC: The dilution QC results were acceptable. Six replicates o f the dilution QC with a pre dilution concentration o f 200 ng/mL, were diluted with water by a factor o f 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 o f 3.0%. See Table 8 for a complete summary o f results. Recovery: Recovery values were calculated by comparison o f 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 o f results. L in earity: The assay was linear for mevalonic acid lactone within the calibration range o f 10.0 ng/mL to 250 ng/mL. Figure 1 shows the 1/X2 weighted least-squares linear regression plot with the actual spiked calibration standard data points for one o f the validation runs. Linearity was also indicated by the correlation coefficients (r) obtained, which were greater than or equal to 0.998 for each o f the first three validation runs. Refer to Table 5 for a summary o f 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 o f six replicates o f 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 o f plasma. 0010 3 418-018:PAGEI-18 G P rim edta Primedica-Worcester Project number. PABX-BVA Page 17 Final Report_____________.____________________ For Primedica Aigus Study Number: 418-018 The variation o f background concentration levels in EDTA rat plasma was evaluated by the analysis o f six unique lots o f plasma during validation Day 2. The result was 8 mean concentration o f 15.8 ng/mL. See Tables 12 for complete results o f the six unique lots o f plasma. Stability: The acceptable criteria for stability evaluations were a mean bias o f <15% from the theoretical values except for the low est calibration, Low-QC and LLOQ QC standards, where the criteria were <20%. Due to the presence o f background concentrating o f 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 o f six control Plasma QC standards. Room temperature matrix stability was evaluated by analyzing six replicates o f 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 o f the results. Freeze/thaw matrix stability was evaluated by analyzing six replicates o f 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 o f the results. Six-day room temperature stability o f final extracts was evaluated by re-analyzing the duplicate calibration curves and the six replicates o f QC standards, in both water and EDTA rat plasma matrices, from the first day o f 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 o f six days when stored at room temperature. See Tables 9 and 10 for the final extract stability results. Studies o f long-term storage stability o f mevalonic acid in matrix at -20C are ongoing. 001074 418-018.-PAGEI-19 G P rimedica 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. 001075 Primedica-Worcester Project number. PABX-BVA 418-018:PAGE 1-20 Page 19 FIGURE 0010i6 Primedica-Worcester Project number PABX-BVA FIGURE 1 Day 1 Validation Calibration Curve 418-018:PAGE 1-21 Page 20 Mevalonic Acid in Rat Plasma Validation Day 1 Peak Area Ratio 0010/7 Primedica-Worcester Project number PABX-BVA 418-018-.PAGE 1-22 Page 21 TABLES 001078 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 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 23.6 70.1 202 8.91 25.0 68.7 200 9.51 25.0 67.8 196 9.38 23.9 68.3 196 9.75 24.0 67.4 195 9.92 23.9 69.8 199 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 24.4 69.9 202 9.29 24.8 68.1 195 10.3 24.1 68.4 195 9.07 23.8 68.2 193 9.10 24.8 68.0 194 10.2 24.5 67.3 195 9.64 6 -3.6% 24.4 6 -2.4% 68.3 6 -2.4% 196 6 - 2.0% 0010<3 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 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 23.9 70.6 10.1 25.2 70.1 9.49 25.4 70.6 9.96 24.6 70.0 9.84 24.8 70.0 NA 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% NA --Not Available. This sample was accidentally spilled during preparation 0010S0 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 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 N om inal 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 M ean Std. Dev. n %RSD % B ias 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% OOlOl Primedica-Worcester Project number. PABX-BVA 418-018:PAGE 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 M ean Resnonse Areas Response Areas 417.5 785 1057.5 1896 3144 4693.5 8433 1066 1981 2963 4975 8678 14279 25380 M ean Std. Dev. % RSD % Recovery 39.2% 39.6% 35.7% 38.1% 36.2% 32.9% 33.2% 36.4% 0.0327 7.4% 0C10o2 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-27 Page 26 TABLE 4 INTERNAL STANDARD EXTRACTION DATA & RECOVERY CALCULATIONS Extracted Standards Solution Standards Mean Resnonse Area Response 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 M ean Std. Dev. % RSD % Recovery 40.9% 39.3% 35.4% 42.0% 38.4% 33.6% 34.5% 37.7% 0.0327 8.7% 00103 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 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 Slone 0.0065514 0.0063508 0.0062253 0.0062800 0.0065156 Intercent 0.0113750 0.0082756 0.0067910 0.0060925 0.0112850 Correlation Coefficient (r ) 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 G 0i0d4 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 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 M ean 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 N om 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 M ean 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. 0U105 Primedica-Worcestcr Project number: PABX-BVA 418-018:PAGE 1-30 Page 29 TABLE 7 SUMMARY OF QC STANDARD FREEZE/THAW (3 CYCLES) STABILITY IN MATRIX Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 M ean Std. D ev n %RSD % Bias WATER QC'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 Nom inal 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. 001086 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 N ominal 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 M ean Std. Dev n %RSD % B ias 39.3 1.16 6 3.0% -1.8% * = D ilution QC's undiluted concentration w as 200 ng/m L (1:5 dilution perform ed p rio r to extraction and analysis) 001037 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-32 Page 31 TABLE 9 6-DAY ROOM TEMPERATURE FINAL EXTRACT STABILITY BACK-CALCULATED CALIBRATION STANDARD CONCENTRATIONS M ean n % B ias 10.0 9.87 10.2 10.0 2 0.4% Theoretical N om 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% GC1088 Pmnedica-Worcester Project number: PABX-BVA 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 M ean Std. Dev. n %RSD % Bias WATER QC'S Theoretical Nominal Concentrations (ng/mL) 10.0 25.0 70.0 200 9.89 243 68.8 198 9.17 25.1 67.7 198 10.5 25.5 68.7 197 9.75 24.4 68 3 200 10.7 24.7 67.7 198 10.1 24.3 69.9 199 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 M ean Std. Dev. n %RSD % B ias 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 OOlOvS Primedica-Worcester Project number PABX-BVA 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 RepS Rep 6 M ean Std. Dev. n %RSD Inter-Assay M ean n Std. Dev. %RSD Concentrations (ng/mL) Dav 1 Dav 2 Dav 3 16.4 16.5 17.0 1 6 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% 0C1090 Primedica-Worccster Project number PABX-BVA 418-018:PAGE 1-35 Page 34 TABLE 12 SUMMARY OF UNIQUE LOTS OF EDTA RAT PLASMA CONCENTRATIONS C oncentrations Cne/mL') L o ti Lot 2 Lot 3 Lot 4 L ot5 Lot 6 11.0 16.2 16.8 16.1 17.6 17.0 M ean Std. Dev. n %RSD 15.8 2.41 6 15.3% ^ X O 'J X Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-36 Page 35 TABLE 13 SUMMARY OF INTERNAL STANDARD PEAK AREAS Low-QC M id-QC H ig h -Q C Water QC's D avi 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 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-37 Page 36 TABLE 13 (Concluded) SUMMARY OF INTERNAL STANDARD PEAK AREAS Low-QC M id-QC H igh-Q C Plasma QC's Dav 1 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 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 M ean Area 4847 Day 2 4503 5363 6736 5184 6350 5493 4091 5157 6063 3722 5601 6044 4445 5474 6088 4231 7133 5471 5397 Dav 3 6100 6151 6067 5909 5159 6755 5413 6040 6238 6216 6049 5746 5985 5811 5615 6084 5879 5467 5927 Comparison o f Plasma OC vs W ater OC M ean Areas % Difference 0.5% 0.5% 4.4% 001033 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 M ean n % Bias % Difference vs W ater QC (T able 1) Dav 2 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 M ean n % Bias % Difference vs W ater QC (T able 1) Theoretical Nominal Concentrations (ng/mL) 25.0 70.0 200 34.5 33.4 32.4 33.0 32.2 33.2 75.7 74.3 75.3 71.8 72.7 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 Nom inal Concentrations (ng/mL) 25.0 70.0 200 34.3 33.2 33.0 33.4 34.8 35.1 76.4 75.9 76.9 75.3 78.3 78.4 198 194 202 194 194 199 34.0 6 36.0% 76.9 6 9.9% 197 6 -1.5% 0.5% 001034 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 Dav3 Theoretical Nom inal Concentrations (ng/mL) 25.0 70.0 200 Rep 1 Rep 2 Rep 3 Rep 4 Rep 5 Rep 6 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 M ean n % Bias % Difference vs W ater QC (Table 1) 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 M ean n % Bias 34.0 18 36.0% 75.7 18 8.1% 196 18 -2.0% 003.095 Primedica-Worcester Project number. PABX-BVA 418-018:PAGE 1-40 Page 39 APPENDIX A LABORATORY METHOD 001096 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-41 Page 40 PH1IMEDICA C O R P O R A T IO N W orcester, M ts u c k u u m Laboratory Method For Analysis of Mevalonic Acid in R at Plaam a By CC/M SD L M N um ber NTVAL00 Revision N um ber 00 Effective D ate N ovem ber 1,2000 P age 1 o f 12 P re p a re d By: ^ Reviewed Bv I * Oste; / ( / / A 7 Date: / / / Date: ---- / i-- j[------ 001097 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-42 Page 41 THE ANAl.VSIS MEVALONIC ACID IN RAT PLASMA BY GC/MSD LM Number: MVALOO Revision Number: 00 Erirctive Daw: November 1 .2TOO Pa* 2 of 12 1 Purpose The purpose o f this Laboratory M ethod (LM) is to describe procedures to accurately determine the concentrations o f mevalonic acid in rat plasma samples by GC/MSD. 2 Scope T he procedures provided in this LM are applicable for the quantification o f m evalonic acid in rat plasma EDTA. During the extraction mevalonic acid (MVA) is converted to mevalonic acid lactone (MVL). The validated calibration range is 10.0 ng/m L to 250 ng/m L o f mevalonic acid lactone in rat plasma EDTA. T h e tcsuIIs can be convened to the uniut o f pm ol/m L o f mevalonic acid in rat plasma EDTA using the conversion described in the calculation section below. 3 Definitions/Abbrcvialions GC: MSD: IS: MVA: MVL: MVL-d: OC: G: Gas Chromatography M ass Selective Detector Internal Standard Mevalonic Acid M evalonic Acid Lactone or DL-Mcvslonk Acid Lactone or M evalonolaclone D L -M ev alo n o lacto n e-4 ,4 ,5 ,5 -d , Quality Control Gravitational Constant4 4 M aterials 4.1 Chemicals DL-Mevakmic acid lactone. Sigma Chemical Co., approximately 97% grade, or equivalent DL-Mcvalonolacione-4 ,4 t5 ,5 -d4 .Cambridge Isolope Laboratories, Inc., 98% grade, or equivalent Ethyl acetate, J.T. Baker, HPLC grade or equivalent Methylene chloride, J.T. Baker, HPLC grade or equivalent 2-Propanol (isopropyl alcohol), EM Science, HPLC grade or equivalent Formic acid, J.T. Baker, reagent grade or equivalent Chloroform, J.T. Baker, HPLC grade ot equivalent Sodium sulfate, Aldrich, reagent grade, or equivalent Deionized water. M illiporc Milli-Q water, or equivalent Helium, Northeast Airgas, Inc., ultra high purity or equivalent Primedica-Woreester Project number PABX-BVA 418-018-.PAGE 1 4 3 Page 42 THE ANALYSIS MEVALONIC ACID IN RAT PLASMA BY GC/M5D LM Number: MVALOV Revision Number: (M Effective Date: November 1,2(I(M Page 3 of 12 lsobutane. Maihcson Gas Products, research grade or equivalent Hexanes, J.T. Baker, HPLC grade or equivalenl Pooled rat blank plasma EDTA 4.2 Consumables Disposable IS mL conical bottom polypropylene lest tubes Ptpetman and tips Eppendorf repeater and tips Disposable polyethylene transfer pipettes or equivalent G lass crim p top autosampler vials with flat bottom inserts or equivalent Polypropylene vials with o-nng sealed caps or equivalent 4.3 Equipment VW R Multi-Tube Vortexcr or equivalent Lab Industries Pipe! Dispenser or equivalent Beckman GS-6 KR centrifuge or equivalent Zymark Turbovap LV Evaporator or equivalent AND ER-182A analytical balance or equivalent JAW GC column, D B -210.30 m, 0.25 nun ID, 0.25 pm film thickneas vr equivalent Hewlett Packard 7683 injector or equivalent Hewlett Packard 6890 gas chromatograph or equivalent Hewlett Packard 5973 mass selective detector or equivalent Hewlett Packard ChcmSuuion (G1701BA, Revision B.01.00) or equivalenl Microsoft Excel 97 version SR-2 or equivalent Note: Equipment, reagents, or consumables can be substituted provided that equivalenl assay perform ance is demonstrated. 5 Procedure This method has been validated for the analysis o f mevalonic acid (as mevalonic acid lactone) in rat plasma EDTA by GCVMSD using 100 pL sample volumes. The validated concentration range is 10.0 ng/m L to 250 ng/mL o f mevalonic acid lactone in rat plasma EDTA. The conversion o f the results to pm ol/m L o f mevalonic acid in rat plasma EDTA is described in the calculations section below. N ote that the calibration curve and quality control standards arc all prepared in water, not plasma. Mevalonic acid is a naturally occurring com pound in plasma and concentrations will vary with diet. Concentrations found m pooled lots o f blank rat plasma EDTA during the m ethod validation were approximately 17 ng/mL. This background concentration level would interfere with the quantitation, especially at th e lower calibration level. This method has been validated to use water matrix calibration curves and quality control samples. The validation demonstrated that the mevalonic acid lactone calculated results for the water matrix is equivalent to rat plasma EDTA. 001039 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-44 Page 43 THE ANALYSIS MEVALONIC ACID IN KAT PLASMA BY GC/MSD LM Number: MVALOO Revision Namber: 60 EITKIiwDitr. November 1,2666 Page 4 of 12 5.1 Preparation o f Reagents Note: Other volumes than those specified may be prepared using the sam e proportions. 5.1.1 Methylene C hloridc:2-Propanol, (90:10, v:v) Solution Combine 900 m l o f methylene chloride with 100 mL o f 2-propanol and mix. Store at room temperature. The expiration d ate for this solution is one week alter preparation. 5.2 Preparation o f Interna) Standard Solutions Concentration calculations are based on assuming that the standard reference materials are 100% pure unless noted otherwise. The following standard preparation scheme is a suggested approach. Appropriate modifications to reach the targeted nominal concentrations are acceptable and will be documented in the study notebook. All standard solutions will be stored at <-20C unless noted otherwise. The expiration dates for these solutions are six months after preparation unless noted otherwise. Ongoing stability studies may change the expiration dates. 5.2.1 Internal Standard Stock (ISA) MVL-d, is a liquid at room temperature and is purchased m vials containing approximately 100 mg. W eigh the vial andMVL-cL- Transfer the MVL-d, to a 50 m L volumetric flask. W eigh the empty vial. Subtract the weights to obtain the amount o f M V L-d, in the volumetric flask. B ring to volum e with Mi)li-Q w ater and mix. The nominal MVL-d, IS concentration o f this solution is 2000 pg/mL. 5.2.2 Internal Standard Spiking Solution tra n sfe r 20.0 pL o f internal standard stock (ISA) into a 100 mL volumetric flask. Dilute to volume with Milli-Q water and mix. The nominal MVL-d, concentration o f this solution is 400 ng/mL. 5.5 Preparation o f Calibration Standard Slock Solutions The following standard preparation scheme is a suggested approach. Appropriate modifications to reach the targeted nominal concentrations arc acceptable and will be documented in the project notebook. For example, if the targeted nominal concentration is not achieved when preparing the analytical calibration slocks, the volum e o f the stock solutions used in subsequent dilutions can be modified so that the nominal calibration standard spiking solution concentrations are achieved. Standard reference materials use assumed to be 100% pure. Standard weights listed are not corrected for purity. All standard solutions will be stored at <-20C unless noted otherwise. Ongoing stability studies are establishing the expiration dates o f these solutions. 5.3.1 M VLPrim ary Stock (A) 001100 \ Primedica-Worcester Project number PABX-BVA 418-018:P A G E I45 Page 44 THE ANALYSIS MEVALONIC ACID IN RAT PLASMA BY CC/MSD l.M Number: MVAL00 Kfvkub Number: M Effertivi Bait: Novembre 2IHHI Page S u f 12 N ote that MV)., is hydroscopic. A large amount o f M V L is used and h is weighed out quickly to m inimize the effects o f absorbing m oisture out o f the air. Store the analytical reference standard under an inert gas such as nitrogen. W eigh out approximately 100 m g o f M VL and transfer it into a 100 mL volumetric flask. Bring to volume with ethyl acetate and mix. The nominal M VL concentration o f this solution is 1000 pg/mL. 5.3.2 Secondary Stock (B) Transfer 3.75 m L o f the Primary Stock (A) into a 25.0 m L volumetric flask. Bring to volume with ethyl acetate and mix. The nominal M VL concentration o f this solution is 150 pg/mL. 5.3.3 Preparation o f Calibration Standard Spiking (CSS) Solutions The following standard preparation scheme is a suggested approach. Appropriate modifications to reach the targeted nominal concentrations are acceptable and will be documented in the study notebook. All standard solutions will be stored at <20"C unless noted otherwise. O ngoing stability studies are establishing the expiration dales o f these solutions. 5.3.4 Prepare CSS solutions in 25.0 m L volumetric flasks as shown in the table below. All solutions are brought to volum e with etby) acetate. Final volumes o f these calibration standard spiking solutions can be adjusted as long as.proportionality is maintained and their preparation is properly documented in the study notebook. Spiking Solution ID CSS-1 CSS-2 CSS-3 CSS-4 CSS-5 CSS- 6 CSS-7 C alibration S tandard Spiking Solution Preparation Stock Solution Stock. Solution Spiking Solution Stock Solution V o lu m e Concentration Final Volume ID (pL) (p g/m L ) (m L) Secondary Slock B 168 150 25.0 Secondary Slock B 337 150 25.0 Secondary Stock B 505 150 25.0 Secondary Stock B 842 150 25.0 Primary Stock A 227 1000 25.0 Primary Stock A 379 1000 25.0 Primary Slock A 631 1000 25.0 Spiking Solution C oncentration (ng/m l.) 1010 2020 3030 5050 9090 15)50 25250 5.3.5 Once prepared, aliquot the Calibration Spiking Solutions (suggested approximately 0 . 6 m l. volumes) into plastic vials with o-ring sealed caps and store them at 5-20C until the lim e o f analysis. 5.4 Preparation o f Calibration Standards (STD) in Water 001101 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-46 Page 45 THil ANALYSIS MEVALONIC ACID IN RAT PI.ASMA BY GC/MSD 1.M Number: MYAluUlf Rrvbion Number: 00 Effredvr Date: November 1,200b Page 6 of 12 C alib ratio n Standard ID STD 1 STD 2 STD 3 STD 4 STD 5 STDb STD 7 Prepare calibration standards on the day o f analysis by combining 2.50 mL o f Milli-Q water, individually, with 0.0250 mL o f the respective Calibration Spiking Solution (CSS) followed by m ixing. T h e final concentration in water is listed in th e table below. W ater V olum e (m l,) 2.50 2.50 2.50 2.50 2.50 2.50 2.50 C alibration Standard Preparation Spiking Solution Spiking Spiking Solution Volum e Added Solution C oncentration (m L) ID (ng/m l,) 0.0250 0.0250 CSS-1 CSS-2 1010 2020 0.0250 CSS-3 3030 0.0250 CSS-4 5050 0.0250 CSS-5 9090 0.0250 CSS-b 15150 0.0250 CSS-7 25250 Kina) Nominal Plaim a Cone. (ng/m L ) 10.0 20.0 30.0 50.0 90.0 150 250 5.5 Preparation o f Quality Control (QC) Stock Solutions The following standard preparation scheme is a suggested approach. Appropriate modifications to reach the targeted nominal concentrations are acceptable and will be docum ented in the project notebook. Standard reference materials are assumed to be 100% pure. Standard weights listed are not corrected for purity. All standard solutions will be stored at -20C unless noted otherwise. Ongoing stability studies are establishing the expiration dates o f these solutions. 5.5.1 Primary QC Stock (AA) N ote that M VL is hydroscopic. A large amount o f M V l, is used and it is weighed out quickly to minimize the effects o f absorbing moisture out o f the air. Store the analytical reference standard uniter an inert gas such as nitrogen. 5.5.2 W eigh out approximately 100 mg o f M VL and transfer it into a 100 mL volumetric flask. Bring to volume with ethyl acetate and mix. The nominal M V l- concentration of this solution is 1000 pg/mL. Secondary QC Stock (BB) Transfer 5.00 m L o f the Primary QC Slock (AA) into a 25.0 mL volumetric flask. Bring to volume with ethyl acetate and mix. The nominal M VL concentration of this solution is 200 pg/mL. 001102 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-47 Page 46 i hi: analysis m ev a lo n ic acid in r a t plasm a by g c/m sd LM Number: MVALOO Revision Number: 00 Ellrflivt D ale November 1,2008 F ife 7 f 12 5.6 Preparation o f Quality Control Spiking Solutions (QCSS) The following standard preparation scheme is a suggested approach. Appropriate modifications to reach the targeted nominal concentrations are acceptable and will be documented in the study notebook. All standard solutions will be stored at -20*0 unless noted otherwise. Ongoing stability studies are establishing the expiration dates o f these solutions. Prepare Q CSS solutions in 25.0 m L volumetric flasks as shown in the table below. All solutions arc brought to volum e with ethyl acetate. Final volumes o f these quality control spiking solutions can be adjusted as long as proportionality is maintained and thenpreparation is properly documented in th e study notebook. Spiking Solution ID QCSS-LOW QCSS-MED QCSS-HJGH QC Standard Spiking Solution Preparation Stock Solution ID Stock Solution Volume Stock Solation Cone. Spildng Solution Final Volume Used Secondary QC Stock BB (Ml-) 3)6 (lig/mL) 200 <mL) 25.0 Secondary QC Stock BB 884 200 25.0 Primary QC Stock AA 505 1000 25.0 Spiking Solution Cone. (ug/iuL) 2525 7070 20200 S.7 Preparation o f Quality Control (QC) Standards in W ater Com bine 100 mL o f M illi-Q water, individually, with 1.00 mL o f each Q uality Control Spiking Solution (QCSS) as shown on the table below. Quality Control Standard ID Low QC Mid QC HighQC Quality Control Standard Preparation Slock Stock Solution Stock Solution Solution Concentration Spiking Volume ID (ng/m L ) (mL) OCSS-LOW 2525 1.00 QCSS-MED 7070 1.00 QCSS-H1GH 20200 1.00 Kina) Volume. (mL) 100 100 100 QC Standard Concentration (ng/mL) 25.0 70.0 200 $.7.1 O nce prepared, aliquot the OC standards (suggested approximately 1.2 mL volumes) into plastic vials with o-nng scaled caps and store them at <-20*C until the time o f analysis. 001103 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-48 Page 47 THE ANALYSIS MEVALONIC ACID IN RAT PLASMA BY GC/MSD LM Number: MVAIJO Revision Number: M Effective Date: November 1,2000 F i l l * of 12 5.8 Sample Extraction 5.8.1 Transfer 100 pL o f each standard (total o f 14), method blanks (in duplicate), control blanks (in duplicate). Quality Control Standards (duplicates at three concentration levels), and study sam ples into individual 15 m L conical bonom polypropylene test tubes. 5.8.2 Add 50.0 pL ofM illi-Q water to each method blank. 5.8.3 Add 50.0 pL o f the Internal Standard Spiking Solution to each tube except for the method blanks. 5.8.4 Vortex the tubes for a minimum o f 5 seconds. 5.8.5 Add 0.100 mL o f formic acid to each tube and vortex for a minimum o f 5 seconds. 5.8.6 Allow tubes to sit on the bench for approximately 30 m inutes to aliuw time for the conversion o f MV A to M V L 5.8.7 Add 0.800 mL o f Milli-Q water to each tube and vortex for a minimum o f 5 seconds. 5.8.8 Add 0.500 m l. o f chloroform to each lube and vortex for a minimum o f 5 seconds. Note: a pipet dispenser may be used to add this reagent. 5.8.9 Centrifuge the tubes approximately 5 minutes at approximately 3500 rpm (approximately 2500 G). Distinct layers should form in each sample tube. Centrifuge again any sample which does not have distinct layers. 5.8.10 Transfer the top (aqueous) layer into new individual 15 m L conical bottom polypropylene test tubes and discard the choroform layer. 5.8.11 Saturate the aqueous layer with sodium sulfate. (Note: approximately 0.15 grams o f sodium sulfate is plenty to saturate the aqueous layer. A measuring spoon may be used to add the sodium sulfate because the exact weight is not critical.) 5.8.12 Add 1.0 m L o f methylene chloride:2-propanol solution to each tube containing the aqueous layer and vortex for a minimum o f 5 seconds. Note: a pipet dispenser may be used to add this reagent. 5.8.13 Centrifuge the tubes approximately 5 minutes at approximately 3500 rpm (approximately 2500 G). 001104 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-49 Page 48 THE ANALYSIS MEVALONIC ACII> IN HAT PLASMA BY GC/MSD 1.M Number: MVAMM Revision Number: OS Effective Date: November 1, 2000 Page 0 of 12 3.8.14 Transfer the bottom (organic) layer into new 15 mL conical bottom polypropylene test lubes. 5.8.) S Repeat the previous three steps. 5.8.16 Evaporate the organic layer to dryness in a Turbovap at 40 5C at approximately lO psi. 5.8.17 Add 50 uL o f the ethyl acetate to each tube and vortex for a minimum o f 5 seconds. 3.8.18 Transfer inlo crimp top autosampler vials with flat bottom inserts. Extracts are stable in autosumpJcr vials stored at room temperature for up to 6 days. Note: If the extracts evaporate to dryness while waiting for analysis they m ay be reconstituted with m ore ethyl acetate before injecting. 5.9 Analytical Conditions GC/MSD analysis is performed as follows: 5.9.1 Oven Column: Initial Temperature: Initial Time: Oven Ramp: Run Time: J& W , D B -210,30 m, 0.25 m m ID, 0.25 pm film thickness 5 0 "C 1.50 minutes Rale Final Temp. Final Time (O m in.) (C) (m in.) 30.0 160 3.50 50.0 230 3.50 approxim ately 14 minutes 5.9.2 Inlet Injection Volume: Rinse Solvent: Mode: Temperature: Purge Flow; Purge Time: Gas Type: Initial Flow: Flow Ramp: Ip L Hexanes Splitless 240 C 50.0 mL/minutc 0.75 minutes Helium 3.5 mL/minutc Constant Flow Mode 001105 Primedica-Worcestcr Project number: PABX-BVA 418-018-.PAGE 1-50 Page 49 THE ANALYSIS MEVALONIC ACID IN RAT PLASMA BY GC/MSI> LM Namber: MVA1XKI k rvioi Number: SO EtTrctive Date: Nevrmbrr 1.2IHH) Pagt* IB of 12 5.9.3 Mass Selective Detector Source: Source M ode Cl Gas: Plow Control Setting: Acquisition Mode: Solvent Delay: 5.9.4 MS Quad: MS Source: MSD Transfer Line: Ions Monitored: Chemical Ionization (Cl) Positive Isobutane 20 Selective Ion M onitoring (SIM ) 6 . 0 minutes (may be adjusted to be as long as possible without eliminating the fust peak of interest) 106-C 250C 250C 5.10 5.11 5.9.4.1 G roup 1: Start time approximately 6.00' Compound Nominal Ion Dwell Retention Time' Tim e1*3 Name MVL (nsk) 131 Im sJ 100 (min.) 7.2 M V L -d, 135 1 0 0 12 1 - Dwell times may be modified to optimize performance. ' = Approximate retention tim e(RT) fo ra new column. Actual RT may vary as th e column is shortened during maintenance. T he RT is confirmed using the isotopically labeled internal standard. 3 = Start tim e may be adjusted. Analytical Run Sequence and Composition Kach analytical run will b e com prised o f bracketing water calibration curves (7 points) each with a water method blank sample (water wiih neither analyte nor internal standard spiked) and a water control blank (water with only internal standard included). Duplicate water QC standards at each o f the three concentrations will be interspersed between the calibration curves. Plasma D ilution Q C's will b e analyzed within analytical batches in which plasma samples arc analyzed at a dilution. T he dilution o f plasma samples and plasma Q C 's will be performed using water as the diluent. Calculations 5.11.1 Chromatograms will be autom atically integrated and visually inspected lo r an acceptable integration using HP ChcmStation (or equivalent). Manual integrations will be performed when necessary. 001106 Primedica-Worcestcr Project number: PABX-BVA 418-018:PAGE 1-51 Page 50 THE a n a l y s i s m e v a l o n ic a c i d jn r a t p l a s m a by c c /m s d LM Number: MVALOO Revbwa Number: 00 ' Effective Dale: November 1, TOM Pug* II uf 11 5 .I1 J Compute the I/X3-weighted, least-squares linear regression relating the peak area ratios (relative to internal standard) o f the calibration standards, to their respective nominal concentration (ng/mL o f M VL in water) using validated Excel spreadsheets (or equivalent). 5.11.3 Using the peak area ratios o f the analyte (relative to the internal standards) in the samples and the appropriate regression equation constants, determine die concentration in ng/m L o f MVL in water o r plasma for all standards and samples using validated Excel spreadsheets (or equivalent). Correct for dilutions if appropriate. 5 .11.4 If requested, the units o f ng/mL o f M VL in w ater or plasm s m ay be convened to pmol/mL o f MV A in water or plasma by multiplying by a unit conversion value {1000 ptnol g/mol ng) and dividing by the molecular w eight o f M VL (130.14 g/mol)- Below is an example showing the conversion o f a sample concentration o f 25 ng/m L o f M VL in plasma. 2 5 .0 ng m l. X lOOOpmol g X lumi = mwt ng 130.14 g 1 9 2 ptnol mL 5.12 Acceptance Criteria 5.12.1 Calibration Standards 5.12.1.) The lower limit o f quantitation (LLOQ) standard's back-calculated concentration must be within 2 0 % o f theoretical nominal concentration. 5.12.1.2 All other standards' back-calculated concentration must be within 15% o f their nominal theoretical concentrations. 5.12.1.3 A minimum of 75% of the calibration standards must meet the backcalculated concentration criteria. (Note: this limit will be rounded down when using 14 calibration standards. Thus, a maximum o f 4 standards may be rejected when analyzing a total o f 14 calibration standards in a batch.) 5.12.1.4 At least one standard at the LLOQ and at the upper limit o f quantitation (Ul-OQ) must meet the back-calculated concentration criteria. 001107 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-52 Page 51 THE ANALYSIS MEVALONIC ACJD IN RAT PLASMA BY GOMSD I.M Number: MVALOO Rrvtstoo Number: M " Effective Bate: November 1 ,2040 Page 12 of 12 5.12.2 Quality Control (QC) Standards 5.12.2.1 The lowest level Q C 's calculated concentration must be within 2 0 % o f the nominal theoretical concentration. 5.12.2.2 Ail other Q C 's calculated concentration must b e w ithin 15% o f their nominal theoretical concentration 5.12.2.3 A minimum o f 6 6 % o f all Q C 's must meet the calculated concentration criteria. 5.12.2.4 At least one QC at each concentration level m ust meet the calculated concentration criteria. 5.12.3 Blanks and Controls A control blank should have a response (peek area ratio) less than or equal to 33% o f the lowest accepted LLOQ calibration itandard The control blanks (in water) should have a response (peak area ratio) <33% o f the lowest accepted LLOQ calibration standard. 5.13 Data Reporting 5.13.1 Sample concentrations less than the lowest calibration standard will be (lagged as Rclow the Quantitation Limit (BQL). 5.13.2 Sam ple concentrations greater than the highest calibration standard will be appropriately diluted with Milli-Q water, re-extracted, and re-analyzed, provided that enough sample volume remains. I f there is insufficient sample available, the original result will be reported as Above the Quunlitation Limit (AQL). 6 Revision History Initial laboratory method. 001108 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-53 Page 52 APPENDIX B REPRESENTATIVE CHROMATOGRAMS FROM VALIDATION DAY 1 C C 1109 Primedica-Worccster Project number. PABX-BVA 418-018:PAGE 1-54 Page 53 Peak Area Report / > fui/S'** Sarapla Kama: 1x Btonk W atsr Notaboo * n afareana: PABX-BVA-1-062 Data Acquired: 10/17/200013.52 O parelD rM A N Data FHa N an a: 0621001.D Data F la Path: D:\MSO 0ATA\PAB\PABX-8VA\1-062-1\ fcuirum ent M attiod N an a: PABX_BVA inetniinant N am e M 57 Inatram ant Vial N um iM r 1 C om pound 1) 2) gmpoynf l N6/1 MVL MVL-d4 TlmtlMlnJ Jan p-- fcAraa i t 0.00 0.00 131 135 0 0 Paafc A rea Rado OiV/OI M* The idlac *ai" Pi Ihm cclumn Indioa f mm manual regiallu i ma partutiuad en ma rampow . P im i Oste and Time: 10/18/200012:39 Paqe 1 001110 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-55 Page 54 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVR\l-062-l\0621001.D Vial: 1 17 Oct 2000 13:52 Operator: MAN lx Blank Hater Inst : MS7 MS Integration Pararne: rteint.p Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTB Integrator) nc:2iaoin j iso i 180 A i 1701 * 160rJ `v , j\ f Sv" 'w ' % J ' w.t. j . , - < - ! I 1 . I j 'n ' i - p ' - r T I I I I I I I . I . I I ! . T - . r . -- < r p - r T - r i I V | n -i I , I I . I , I I . | . . r . - r . | . . inm-> aio aao 6.30 a..a,so aao 8.70 aao bjo 700 rio ran r.ao 740 r.so 7.ao 7.70 7.ao 7.90 MVL .......................... Amount: Not Found Ion: 131 (0.000) l\ V.yv'',.yi J , Ktt 64fl Iaao I J 1 1 1 , 1 1 1 a a.ito a.ao r d4 Ion: 135 (0.000) b.70 aao jbo f 00r 7.10 'r a o.... t 5 o 7.iw i a 7.ao 7.70 7.ao~r y jo __________ Amount: Not Found _________________ 1\ I \ ... / V f r * - n -r -J ----- 6.20 6.30 6 4 0 6 50 6 60 6.70 6.80 6.90 7.00 710 7.20 730 740 7.80 7.60 7,70 . 7.80 780 0621001.D PABX BVA.M Wed Oct IB 12:40:03 2000 Page 2 001111 Primedica-Worcester Project number; PABX-BVA 418-018:PAGE 1-56 Page 55 Peak Area Report Sampia N am *: 1x SW-1 W ataf Notabeni! Ratawnoa: PABX-8VA-1-082 D a li A cq u a ta: 1Q717OOOi :11 O p erato r MAN Data H a Nam e: 0621002.D Data F t Path: 0 :W S 0 OATAVPAB\PA8X-BVA\1-062-11 tnatruiiw nt M attw tf N an a: PABX_BVA In stium eat Kam a: MS7 tn a tn a m at V lalN uo ifaar 2 Compound f 1) 2) Compound Naaw MVL MVL-4 BatesILtai pmtiwmj MS 7.17 Isa 131 135 ssSj b c 250 3260 p--*1Af--.Rate 762.2E-4 M* Th lari* "m**in H*wccli*tv> inmcam that mr nri inagnW an-- pmrtanm * an Biol etm yuund. Print Oat* and Tim -107102000 12:40 P*gn 1 001112 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\FAB\PABX-BVA\l-062-l\0621002.D Vial: 2 17 0ct~2000 14:11 Operator: MAN lx Std-1 Water Inst : MS7 MS Integration Parana: rteint.p Quant Method : D:\HPCHEM\1\MBTHDDS\PABX_BVA.M (RTE Integrator) jiM ir j 1500 ........... TTC: 06210Q2.D A / 1000 500 r m - > aio 6 a MVL I11.t'i'!Im r-' "-I'. I T " 1!'" , II. Ir u IJ . I II. 36 50 6.80 6 70 660 690 700 T IP 7.20 7.30 7.40 7.50 760 7 70 760 7.90 Amount : 0.00 0621002.D PABX_BVA.M Wed Oct 18 12:40:22 2000 Page 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-58 Page 57 Peak Area Report Sam ple Name: 1x Std-2 WMr Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/170000 14:30 Operator: MAN Data FHe Name: 0621003.0 D ata FHe Path: D:\MSD_DATA\PA8\PABX-BVA\1-062.1\ tnatnim ent Method Name: PA B XJVA tnatnim cnt N a ta : MS7 Inatniroent Viet N um ber 3 Compound 1) 2) MVL MVL-4 M nbon Tim e IM kU 7.14 7.17 Is o 131 135 P w K A rw IC 587 at 3064 eaM LA M ftJM to 1423 fcr a The letter ~m ~ ki M acellam i InillBtlae tha manuel I Pnm Date and Time: 10/18/2000 1240 Pap* 1 001114 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-59 Page 58 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\HSD_DATA\PAB\PAHX-BVA\1-062-1\0621003.D Vial: 3 17 Oct 2000 14:30 Operator: MAN lx Std-2 Hater Inst : MS7 MS Integration Pararne: rteint.p Quant Method : D:\HPCHEM\1\MBTH0DS\PABX_BVA.M (RTF Integrator) f^ isss 1500 1000 TIC: 08210C3 11 II 900 T. ,r* r , t fflma 0.10 6.20 630 6.40 * 6J0 11!1" ' ' 11' M" I1- rp .^1T.-n , .. ............ I.II --n - T -- 6.00 6.70 ftBO 8JO T.00 7 1 0 720 7.30 7.40 7S0 7.00 7.70 7.00 730 MVL Amount: 0.00 Ion: 31 (7.144) Reap: 567 ! 1 I ...............I - T -- I - I - ' r T n ' . - ' | T . - n . ^ . . T . , ' I T ] ' . T r r p T T Y T i 'r 1 ' 1 1 1 | ' 1 1 I ' 1 1 | 1 ' ' ' I ' ' 6.20 030 640 600 660 0.70 6 0 0 6.00 7.00 7,10 720 7.30 740 7.50 7 6 0 7.70 7.00 7JC P I-d4 Ion: 135 (7.171) Amount: 0.00 Reap: 3984 A iv ! -- I ...... I ' ' 1' I'1' '--1' 1I --' I ' ; I . I . I--I ' . I 11I I .................... ..-- T, - r . . ! 6.20 6.30 640 6.50 6.00 6 7 0 ...6 6 0 _ 6 90 7,00 7.10 7.20 720 7 40 7.50 7,60 7 70 7.00 7.90 0621003.D PABX BVA.M Wed Oct 18 12:40:41 2000 Page 2 001115 Priinedica-Worcester Project number PABX-BVA 418-018:PAGE 1-60 Page 59 Peak Area Report Samp i* N an a: 1 Std-3 W atar Notebook Ratefanca: PABX-BVA-1-062 Date A cquim i: 10/17/2000 14:50 Operator; MAN Data FUa N an a: 0621004.D Date FHa Patti: D:1MSD_0ATA\PA8\PABX-BVA\1-062-1\ Instrum ent Method Name: PABX_BVA te a m n a n t Nai a: MS7 Instrum ent V ia l Num ber: 4 Com pound* 1) 2) Com pound Name MVl MVL.-04 Retention Tim a IMto.1 7.15 7.17 (an 131 135 P tP k K 901 SI 4234 Peak Area Ratio 212.8E-3 m* The toflo V tnthl>coturon lraho*M thatimwi< kitey eflcnw performed on ft P im i Date and Time: 10/18/200012:40 Page 1 001116 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_nATA\PAB\PABX-BVA\l-062-l\0621004.D Vial: 4 17 Oct 2000 14:50 Operator: MAN lx Std-3 Hater Inst : MS7 MS Integration Parents: rteint.p Quant Method : D:\HPCHEM\l\MBTHODS\PABX_BVA.M CRTS Integrator) jAbundme* 2000 TIC: 06210040 lOOOj / \V I y-T'.-T I, , . i-n I I I I P ' l ! T | i' < T i ; r r u | , , ! I p i 'i'H i rri | V . T T y .,.,. fan - 610 420 6.30 6.40 6.50 6.60 6.70 6.60 6 J0 7.00 7.10 73P MVL Amount: Ion: 131 (7.146) Resp: 901 a /i V . I IV | i , , , , i , - r 'l - ' > , < > p i . i , , -r 7.30 7 40 7.50 7.60 7.70 7.60 7 90 0.00 T 1 11...~n- 620 6.30 640 6.60 6.60 6.70 6.80 6.90 7.00 7.10 7 . 7.30 7.40 7.6S0 7 6 0 7.70 7.60 760 d4 ....''_____________ Amount: 0.00 Ion: 135 57.169) i Resp: 4234 A ....... ....._ L l 6.20 6.30 6.40 6.50 66 0 6 70 6.80 6.90 700 7.10 733 7.30 7.40 7.50 7.60 7.70 7.80 7.90 0621004.D PABX_BVA.M Wed Oct 18 12:41:01 2000 Page 2 001117 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-62 Page 61 Peak Area Report Sample Name: 1Std-4 Water Notebook Raterenca: PABX-BVA-1-062 Date Acqukad: 10/17/200015:0 Operator: MAN Data Fite Nam e: 0621005.0 D ate File Path: D:\MSD_DATAVPABPABX-BVAV1-062-11 btetrum eni M ethod Nam e: PABX_BVA Inatniinant Name: MS7 tnstnim ent V ial N um ber 5 Compound 1) 2) Compound Nama MVL MVL-d4 RaftMrtton Ttm alM tn.1 7.14 7.17 Joq 131 135 Peak A iaa Ifr 1285 3748 P w hftm R stft. 342.SE-3 M*TN laWafHi"HiWtcoteau Indtcaf l matm--!mtogratWrwopmformedonthat coiapuund Print Date and Time: 10/18/20001241 Pt|e 1 0G1118 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 Hater Inst : MS7 MS Integration Params: rteint.p Quant Method : D:\HPCHEM\1\METB0DS\PABX_BVA.M (RTB Integrator) Hbndw* TIC: 0621005.D 1500 ! 1 1000 500 T7-> 6.10 MVL 6.20 rT r 6 . - . . - i - r T i T T r .. ........................................ I * ............................ . . . I , . il ' . Ti 6 4 0 ...6 5 0 6.60 6.70 6.80 6 90 7.00 7.10 7.20 7 3 0 740 7.50 Amount: 0.00 7.60 7,70 7.80 7.90 I' ion: 131 (7.143) !Reap: 1285 T1 8.20 4 30 6.40 8.50 860 6.70 6.80 6.90 _ -d4 Ion: 135 (7.171) ......... ... IResp: 3749 .T rt-t l - p r 1t ------ - : 7.00 7.10 7.20 7 . Amount: r ' ' i 1' 1 i 111i i ' ' , 1i 1i 1 7.40 7.50 7.80 7.7p 7.80 780 0.00 ' .............. :AI \ i 6.20 6.30 6.40 ' 6.50 6 60 6.70 680 690 7.00 ' 7.10 7.20 7 30 7.40 7.50 7.60 ` 7.70 7 .a T * 7.90 0621005.D PABX BVA.M Wed Oct 18 12:41:20 2000 Page 2 O G llis Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-64 Page 63 Peak Area Report Sample N an a: tx SW-5 W ater Notebook R eteienca: PABX-eVA-1-002 Data Acquired: 10/17/200015:29 Operator: MAN D ata File N aan; 0621006.D Data FHe Patti: D:\MSD_DATA\PAB1PABX-8VAt1-062-1\ Instrum ent Method Name: PABX_BVA Inetum ant Name: MS7 tnatrum ant Vial N uaiber 6 9fflBWlrti 1) 2) CompoundNnn MVL MVLd4 IT Th M tar "m** NI-Ms--D--M TVwJWal 7.1 7.17 iss 131 135 Peak Area 2208 3603 Peak A rea Ratio 5B7.BE-3 Print Data end Time: 10/18/20001241 Page 1 001120 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-65 Page 64 Quantitation Report (QT Reviewed) Data Pile Acq On Sanale Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621006.D Vial: 6 17 Oct 2000 15:29 Operator: MAN lx Std-5 Water Inst : MS7 MS Integration Pararos: rteint.p Quant Method : D:\HPCHBM\l\MBTHODS\PABX_BVA.M (RTE Integrator) i 1900| TIC:0621006i) 1000* I\ I .'i-."1ir. IIII'!' n n -> 6.10 6.20 6 .X 6.40 6.30 MVL ; Ion: 131 (7.1445 i!Reap: 2208 i i v ...... r '-- 1-7'1'M " " I1" 1 I' ir.-|VTI I...... IIi iiI! iI I ....I IIIiIP' I r 6,60 6.70 8.80 6 30 7.00 7.10 7.20 730 7 40 7.S0 7.68 7.70 7 790 Amount: 0.00 ............ . ; ...... ........ . A II n VV T' I 620 6.30 6.40 6 50 660 6.70 6.80 6.80 7.0 7.10 7.20 7.30 740 7.90 7.M ' 7 60 730 d4 Amount: 0.00 t Ion: 135 (7.172) .... ........... Reap: 3693 A I\ /V n- 620 6.30 6.40 6.50 '~6.60 6.70 ' 6.C0 6.90 7.00 7.10 7.20 730 740 7.SP 7.60 7 70 7 60 7 90 0621006.D PABX-BVA.M Wed Oct 18 12:41:39 2000 Page 2 001121 Primedica-Worcester Project number PABX-BVA 418-018.PAGE 1-66 Page 65 Peak Area Report Sample Name: lx SM -6 W ater Notebook Reference: PABX-BVA-1-082 Date A cquired: 10717000015:46 O p erator MAN Date F ie Name: 0621007.0 Data File Path: O:\MSO_OATA\PABiPABX4VA11J62.1V Inabum ent Method Name: PABX_BVA Instnim ent Nam e: MS7 Instrum ent V ia l N um ber 7 Com pound* 1) 2) Compound Name MVL MVL-04 R d ifitiM Tim efM tn.) 7.14 7.17 !ss 131 135 P it i Am t 4054 m 4062 Peek Aree Retto 99&0E-3 PM itD ete and Time: 1W16I2000 12:41 Page 1 00112J2 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-67 Page 66 Quantitation Report (QT Reviewed) Data File Acq On Sanale Mise D:\MSD DATA\PAB\PABX-BVA\l-062-l\0621007.D Vial: 7 17 Oct-2000 15:48 Operator: KAN lx Std-6 Water Inst : MS7 MS Integration Parame: rteint.p Quant Method : D:\h p CHEM\1\m e t h ODS\PABX_b v a .m (RTE Integrator) ~TW f0621007.0..... 2000 f;\i 1500 II1000 500 V r> .. r 1 " :1 .I Ir-i |-rr. .,..j, rf 11-1T-fl-T'.iyr-ri"J r'r 6,10 420 6 30 6.40 650 8.60 &7D 6 60 6.90 7 J0 7 10 7.20 7.30 7,40 7.50 7.B0 7.70 7.60 730 MVL ___________ ______________________ Amount : 0.00 i Io n : 1 3 1 (7 .1 4 3 ) ! Resp: 4054 A 6.506.20 6 . 6.40 *' *T '*" r r iT r. . v r - r i - n ' N T ' - p ^ r . ' r r , ,r .V > 6.60 6.70 6.60 6.90 7JO 7,10 720 730 7.40 7X0 760 7.70 7.60 790 d4 Amount : 0.00 Ion: 135 (7.170) ; Resp: 4062 I. !\ ..J.L 6.20 6 3 0 6.40 6 50 6.60 6.70 880 6.90 7.00 7.10 7 .2 0 f 7.30 7 40 7.50 7.60 7.70 ' 7.60 ' 7.90 0621007.D PABX BVA.M Wed Oct 18 12:41:59 2000 Page 2 001123 Primedica-Worcester Project number PABX-BVA 418-018'.PAGE 1-68 Page 67 Peak Area Report Samp* N am *: 1 S U -7 Water Notebook Retarenee: PABX-BVA-1-0S2 Date Acqubad: 10/17/200016:08 O perator MAN D ata FUa Name: 0621008,D Data FUe Patti: D:\MSD_DATAIPAB1PABX-BVAV14)62-1 \ biatruatant Method Name: PA8X_BVA Instrum ent Name: MS7 In atrum entV IalN uaibar t C om pound* 1) 2) Compound Name MVL MVL-4 T im * (Mtn.) (os 7.14 131 717 13S P-- kA raa ( 6314 3766 Peak Area Ratio 1 8 7 7 E -2 W Th* l*W f "m" In OH column Indicstoo that marni l i kiioqrohon CP-- porTcrmcd on Slot m u --pu lp]. P rint Data and Time: 10/18/2000 1242 Page 1 001124 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-69 Page 68 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621008.D Vial: S 17 Oct 2000 16:08 Operator: MAN lx Std-7 Hater Inst : MS7 MS Integration Parama: rteint.p Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC: 0621006.0 2000a 10001 ! i'i... I.....I" *.... :' 810 620 6,30 8 . 660 8.60 6.70 0 00 M V L ____________________ 'Ion:' 131 (7.141) Resp: 6314 Amount:______ 0.00 ( I I i _______ j E -yr-8-r,. .,,T "' 1 ' !''>'T... ;iH 'I i' Ti'{ 6,20 's a o 640 6.50 6.50 6.70 8.80 6.80 .......... 7 10 7,20 7 . 7.40 7SD 7 7.70 7.80 7.80......... -d 4 .... Amount:" .00 Ion: 135" (7.169) Resp: 3766 r - r - r r - r - r - , n -*a| . , . i f t - v r - r T ' r ' , .r- ^ .8.20....6.30.....6 ,4 0 ..6 5 0 .....6,60. 670 6 60 690 7.00 _ 7.10 7.20 7 30 7.40 7 60 7.80 7.70 7.80 7.90 | i 0621008.D PABX BVA.M Wed Oct 16 12:42:18 2000 Page 2 001125 Primedica-Worcester Project number: PABX-BVA 418-018.PAGE 1-70 Page 69 Peak Area Report Sam p i* N am *: lx Control W an t Notebook Re m onco: PABX-8VA-1-062 Date Acquired: 10/17/200016:77 O ponter: MAN D ate R io Nomo: 0621009.D D ate FU* Patti: 0 :1MS0_ATA1PAB'PABX-BVA\1-062-1 \ Inatnim ant Mattiod Nam *: PABX.BVA trntnm w nt Mama: MS7 kM trum ont V ial N um ber 9 C om eound# 1) 2) CemDOund Name MVL MVL-04 AJn M-n- w-n- Tlm*(Mtn,l ooo 7.17 ISB 13t 135 ***-- MI 0 4595 Peak A rea Rado 000.0E-3 M* Ttw Utter "m" m on column P rint Date and Tim e. 10/18/200012:42 Pape 0C11Z6 Primedica-Worcester Project number PABX-BVA 418-018-.PAGE 1-71 Page 70 Quantitation Report (QT Reviewed) Data File Acq On Sanale Mise D:\HSD DATA\PAB\PABX-BVA\l-062-l\0621009.D Vial: 9 17 Oct~2000 16:27 Operator: MAH lx Control Hater Inst : MS7 MS Integration Params: rteint.p Quant Method : D:\HPCTEM\l\METHODS\PABX_BVA.M (RTE Integrator) &nMK* 2000 1500 !!i 1000 TIC: 0621009.D ift\ i 500 !_ !i'i " ,1.11111 M I. 1111 n 111.fi!.y ,, 11,,, 'I''''" 'in in--' 'I'.'^ -'1-- p ITune- 6.10 6.20 6.30 6.40 6.50 6 60 6.70 660 6.90 TOO 7.10 72P 730 7,40 7.50 7 60 7.70 7 BO T J MVL Amount: Wot Found f" Ion: 131 (0.000) A y\ v r\ 6.20 6 30 6.40 630 660 L-d4 m Ion: 135 (7.170) !Reap: 4595 6.70 I . | i "f f t-l I i-l I ' I I 'I ' I I ' I ' ' | i ' r T T Y T -rr I . I I I I I I I ; I I .6 6 0 7.00 7.10 7.20 7.30 7 40 7 80 7 60 7 70 7.80 760 Amount : 6.00 n 1 6 . 6 30 640 6.50. 6.60 6.70 6.80 660 700 7.10 720 7.30 7 40 760 ' 7 60 7 TO 760 ' fa iT 0621009.D PABX BVA.M Wed Oct 18 12:42:37 2000 Page 2 001127 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-72 Page 71 Peak Area Report Sam ple Nam a: lx OC HIGH W elw Notebook R etw anea: PABX-BVA-1-062 Data Acquired: 10/17/200018:47 Operator: MAN Data File Name: 0621010.D D ata File Path: DAMSO_DATA\PABV>A8X-BVA\1-062-1\ Instrum ent Method Nam a: PABXJ9VA Inatnim aat Noma: MS7 Instrum ent V ial Number: 10 Com pound* 1) 2) Compound Nama MVL WVL-d4 PnMatIMuVtrUNlmtt Tlm e(M ln.l 7.14 7.17 fe a 131 135 Peat Area { 5386 4021 Peak Area Ratio 133.4E-2 M* H ie laMar ~wi~ In tW co harm Indteetas that annual mtay ahon mas panorami en U t sempeund. P itrt Data and Time: 10/18/2000 12:42 Page 1 ^01128 Primedica-Worcester Project number PABX-BVA 418-018-.PAGE 1-73 Page 72 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\M5D_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 Pararne; RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) SKindiinc' TIC: 0821010.0 2000 1000 I 6.10 8-20 8 X MVL I '[ I' - ' l 8.40 6.60 6.60 6.70 '' ~ r...... :' .1-\ rrrt .|....r,.i.....|im'| .i.iiIIfrr 6,60 8.90 TOO 7.10 T.20 730 7.40 7.30 760 770 760 7J90 Amount: 0.00 0621010.D PABX BVA.M Wed Oct 18 12:42:56 2000 Page 2 GG1JU9 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-74 Page 73 Peak Area Report Sample N am 1x OC MID W ater Notebook R aftranca: PABX-6VA-1-062 M e Acquired: 10(17/200017:07 O p erator MAN Date File Natna: 0821011.0 Data File Path: D:\MSO_DATA\PAB\PA8X-BVM1-062-1\ Instrum ent Method Name: PABX BVA Instrum ent Name: MS7 Instrum ent V M N um ber 11 C om pound* 1) 2) Compound Name MVL MVL*d4 7.14 7.17 !aa E s s k A ^ e t FeekA m afiaillo 131 2034 m 47U.5E-3 135 4323 M *Thalartar*W *inauce <mtin in e te a ra tlh a tmeiiiwH n ta g isb o a a s pertnm iar taiithetcBwmQlind. Prim Date and Time: 10/18/2000 12:43 Pad 1 001130 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: MAM IX QC MID Water Inst : MS7 MS Integration Params: RTBIHT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTB Integrator) ^Untano 2000 TIC:0B21011.0 iA\ l\ ;I ism - I 1000' // I; ! * i 1/ \ I ! y.T. I1 I-I, I n IITT F> I I Iprm i, ,* / V. fi''I ii| IIl'P1iiii ; ' f m -n * *'!< 1 IM .........,i iii W - > g.10 6.20 S.30 6.40 6.S0 6.60 6.70 6 J0 6.90 7.00 7.10 7JD 7JO 7.40 7.SO 7.60 7.70 7.B0 700 MVL________ _____ __________________ Amoun t ".0 " " ______ Ion: 131 T7.42 ................ ....... . Reap: 2034 A / s a i a 640 6 50 6.60 -d4 Ion: 135 (7.169) Reap: 4323 ' I ...........r v ` 6.70 6.80 JA , l . . . , . r .r .r , . , . ,, 6.90 7.00 7.10 730 730 Amount: 7.B ' ' 7 3 " 7.60 7.70 7.M 0.00 7.90 1 I 670 6.30 6 40 W I\ 'I'' 660 6.70 T a p 6.90 ' 7.W 7.10 ' 7.2) 7.30 7.40 7:50 7.80 7.70 7.80 7.90 I II 0621011.D PABX BVA.M Wed Oct 18 12:43:16 2000 Page 2 001131 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-76 Page 75 Peak Area Report Sam ple Nam: 1x OC LOW Water N o te b o o k R f re n a : PABX-8 VA.1-062 D ate Acquired: 10/17000017 26 O p erato r MAN D ata FBe Name: 0621012D D M FUa Patti: D:\MSD_OATA\PAEnPAaX-BVA\1-Oe2.1\ Instrum ent M ethod Name: PABX_BVA tnmlrument Name: MS7 Instrum ent V ial N um ber 12 Compound 1) 2) C9111BM9VVLnaH6IM MVL-tM T irw tW in J la s 7.14 131 7.17 135 P-- A rea tC 680 m 4096 P ttlK l ~ T p *t" 106.06J k r The le tte r "m " in Se cokown inditiatm that n w e iim yf Hen tm patlennad on Siat cooipoiind. Print D ale and Time: 10/16/2000 12:43 Page 1 001132 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-77 Page 76 Quaaeication Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSDJDlATA\PAB\PABX-BVA\l-062-l\06210l2.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 (RTE Integrator) 0621012.D PABX BVA.M Wed Oct 18 12:43:36 2000 Page 2 001133 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-78 Page 77 Peak Area Report Sam ple Name: lx QC LLOQ W at Notebook M m m c * ; PABX-BVA-1-062 D e li Acquirap: 10/170000 17:46 O perator: MAN D a ta n te Name; 0 6210130 Oata File P atte O:W S0_0ATA\PA8\PABX-BVA\1-002-1\ Instrum ent M ettiod Name: PABX-BVA Instrum ent Naara: MS7 kw tram ent Vial Number: 13 C om oeund# 1) 2) Compound Name MVl MVL-4 nVmtOQHm - i -- -- Tim e (M ia i 7 .t4 7.17 )S 0 131 135 EnfcAas IC 354 4755 Peak Aree Ratto 744.5E-4 M* The letter I n " in Hits column M t e a t u that manual M egrallenw es performed m diet compound. P rint D a n and tim e'. 10/18/2000 12:43 Page 1 001134 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\l-062-l\0621013.D Vial: 13 17 Oct-2000 17:46 Operator: MAN lx QC IiLOQ Water Inst : MS7 MS Integration Pararne: RTBINT.P Quant Method : D:\HPCHBM\l\MBTHODS\PABX_BVA.M (RTE Integrator) ^taunkna 2000 TIC: Q621013.D A 1800 1000 I V rne-o.. 6.Q 8.20 6.30 .40 ' g - MVL _ _ _ _ _ _ i ion: 31 (7.138) Resp: 354 I i s 6.70 a a o ' eso 'rb o 7 . ' t W Amount: !I\\ t q ' ' m T J p ' r p ' r.7 o " r & ` r io 0.00 / \!... ________ ____________________ ____ T " ---1 I 'n I 1 1 . I I _.l 6.20 6.16.20 6.30 640 a . 8.80 8.70 680 6.90 7.00 7.10 T.20 7 30 7.40 730 730 " iT O Amount: 0.00 " IIcojnU:. 135 (7.170) iResp: 4755 780 7.90 i\ i\ )V i_820 830 6.40 6.50 8.80 6.70 6.80 6.90 7 .0 0 ... 7.10 7 3D ....7.30 7.40 7 30 7 60 7.70 7 t 7.90 0621013.D PABX BVA.M Wed Oct 18 12:43:56 2000 Page 2 001135 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-80 Page 79 Peak Area Report 8am pte Nanw: 1x QC HIGH Pteama Notebook Watetanoa: PABX-BVA-1-062 Date A cq u im i: 101712000 18:05 Operator: MAN Date Fite N aim : 0021014.0 Date Fite Pate: D:\MS0_0ATA\PAB\PABX-8VA\1-062-1\ Instrum ent Method Nama: PABX_BVA teabum ent Nama: MS7 hn bum ant V M N um ber 14 Compound 1) 2) Compound Nama M Vi. MVL-04 Tim a (M in.) an 7 .1 4 131 7 .1 0 135 p--HA-- !E PillArmWW 8537 130.5E-2 6540 H T -T tie ien w 'ln -lne ilscp H m im n d igu te m a tiian u elin lepraMen'iwepsrt-- 4 " -- -- d. PnM Data and Tune: 1018/2000 12:44 Pag ). QG1136 Primedica-Worcester Project number: PABX-BVA 418-018 :PAGE 1-81 Page 80 Quantitation Report (QT Reviewed) Data File .Acq On Sample i Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621014.D Vial: 14 17 Oct 2000 18:05 Operator: MAN lx QC HIGH Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Mundane 4000; 3000; nC:0621014.D !\ i\ !\ 2000; iooo; I !T r-> 8 10 6.20 6 .X 640 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 ~ ~ Amount: .0 _ _________ : Ion : 131 (7.137) i R e s p : 8537 A !\ I o.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.S0 7.60 7 70 7,60 7 90 'rL-d4 Amount :______ 0.00_____________~ m ion: 135 (7.161)..... ; Resp: 6540 !\ I\ ;v 1-- ,----- 6 20 6.30 6 40 6.50 6 60 6.70 6.80 6.90 7.00 710 7.20 7.30 740 7 50 7.80 7.70 7.80 7 90 0621014 .D PABX BVA.M Wed Oct 18 12:44:15 2000 Page 2 001137 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-82 P a g e 81 Peak Area Report SampleName: 1xQC MID Plasma Notebook Reference:PABX-BVA-1-062 DateAcquired: 10/17/2000 18.25 Operator MAN DataFiteName: 0621015.D InstrumentName: MS7 DataFilePath:O:\MSD_DATA1PAB\PABX-BVA\1-062-1\ InstrumentVialNumber: 15 InstrumentMethod Name: PA8X_BVA Compound f 1/ 2) Comoound Name MVL MVL-4 Retention Time (Mini Ion 7.14 131 7.18 135 PeakAma M* 2024 3988 Peak Ab a Ratio 507.5E-3 M* * The letter V tn this column Indicele that menuet inteqretion was pertormod on that compound. Pnnt Dale end Time: 10/18/2000 12 44 Page 1 0C1138 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\l-062-l\0621015.D Vial: 15 17 Occ 2000 18:25 Operator: MAN lx QC MID Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundance 20Q0J T1C.0621015.D 1500 1000 sool V T . V P " - .............. .. : . '' T M- I , ............ ...... " . * * 1 I1 ' ' 1 I 1 1 1 1 I 11 6.10 6.20 6.30 6.40 6.S0 6.60 6.70 6,80 6.90 7.00 7.10 7.20 7 . 7.40 7 .6O 7.60 7.70 7.80 7.90 MVL Amount: 0.00 Ion:' 131 (7.137) Resp: 2024 A i 1i \ \ 630 6 .X 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 [VL-d4 Amount: 0.00 ' Ion: 135 (7.165) Resp: 3988 !\ !\ !\ 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 0621015.D PABX BVA.M Wed Oct 18 12:44:34 2000 Page 2 001139 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-84 Page 83 Peak Area Report Sample Name: lxQC LOW Plasma Notebook Reference:PABX-6VA-1-062 DateAcquired: 10/17/200016 44 Operator: MAN DataFUaName: 0621016.D InstrumentName: MS7 DataFHs Path:D:NMS0_0ATA\PA8V>A8X-BVA\1-062-1\ InstrumentVialNumber: 16 InstrumentMethod Name: PABX_BVA ComDOund # 1) 2) w&'in<iwm MVL MVL-44 Retention Time (Min.) &Q 7 14 131 7.16 135 PeekAran W 1115 m 4700 Peak Araa Ratio 237.2E-3 M* Tit icrte r'n T in this column indicates that manual integration teas performed on that compound. Pnnt Date and Time. 10/18/2000 12:44 tag? ) 001140 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-85 Page 84 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-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\l\METHODS\PABX_BVA.M (RTE Integrator) j^toundanc* i : 2000 TIC: 0621016D A 1500 1000 ! 500 n Tim-> S to 6.20 6.30 6.40 6 80 6 0 STO 6.80 6 90 7.00 7 1 0 7.20 7 30 7.40 7 50 7.60 7.70 7 80 7 90 m v L_____________________________________ Amount: .0 r i o n : 131 (7.139) I Resp: 1115 I I..... _ A . 6o.20 6 30 6.40 6.50 6.60 6 70 6.80 6.90 7.00 7 1 0 730 7.30 7 40 7 .50 7.60___7_70 L-d4 __"_______________Amount :______ 0 .0 0 ____ Ion: 135 (7.162) : Resp: 4700 : !\ \ i\ 7M 7.90 6.20 6.30 6.40 6.50 6.60 6 70 660 6 90 7 00 710 _ 7.20 7.30 7.40 7 50 7 60 7 70 7 60 7 90 0621016.D PABX BVA.M Wed Oct 18 12:44:54 2000 Page 2 001141 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-86 P age 85 Peak Area Report Sample Name: 1ControlPlasmi Notebook Reference:PABX-BVA-1-062 DateAcquired: 10/1712000 19:04 Operator: MAN Date FUe Name: 0621017.D InstrumentName: MS7 DataFilePatti:D:\MSD_DATA\PAB\PABX-BVA\1-062-11 InstrumentVialNumber: 17 InstrumentMethod Nome: PABX_BVA Comoourtd # h 2) Comoound Name MVL MVL-4 Retention Time fMtn.) Ion 7.14 131 717 135 Peak Ama IT 577 m 4860 Peak Area Ratio 118.7E-3 M* > The lener *nT In this column Indiceu s that manual integration was pertotmed on that compound. Print Date and 1ime: 10/18/200012:45 Page 1 001142 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-87 Page 86 Quantitation Report (QT Reviewed) Data Pile 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 Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) (tbundtnce ! 2000: TIC:0621017.0r 1r.1 n j 1500: iv 11 J 1000i j\ sool / i __ / r '~T^ 1 r'; ' r'" 1 ,1'n f7*!r* hm-- 610 620 6.30 6.40 6.50 6.60 6.70 6.90 6.90 7.00 7.10 7.20 7.30 740 7.50 7.60 770 7.80 7.90 MVL Amount : 0.00 i Ion: 131 (7.138) | Resp: 577 A ;\ i I) I !\ i i\ i i I 1 i 1 ! j 1 670 630 640 6.50 660 6.70 6.80 6.90 7.00 7.10 7.20 7.30 740 750 7,60 770 7,60 7.90 X-d4 ~ ... Amount: 0.00 m Ion: 135 (7.165) Resp: 4860 A 6.20 6.30 640 '6.50''6.60 6.70 6.80 6.90 7M !\ ;V --T"" 710 7.20 7.30 7.40 7.50 7.60 7.70 780 7.90 0621017.D PABX BVA.M Wed Oct 18 12:45:13 2000 Page 2 001143 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-88 Page 87 Peak Area Report Sample Nam: 2 *OC HIGHWater Notebook Reference:PABX-BVA-1-062 DateAcquired: 10/17/200019:24 Operator: MAN DataFileName: 0621018.0 InstrumentName: MS7 Data FUePatti:D:\MSD_DATA\PA8\PABX-BVA11-062-1\ InstrumentVialNumber: 18 InstrumentMethod Name: PABX_BVA Compound!? 1) 2) Compound Name MVL MVL-04 Retention ThnelMin.) Iso 7.14 131 7.17 135 pensais M: 7314 5545 per*Area Retto 131.96.2 M* Tha tettar V in th ti column indicate that manual Iniapration w n performed an that compound. Print Date and Time: 10/18/2000 12:45 Pant 1 001144 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) TIC:062116.D f\ |t il3000; 1 12000j ii 1000- ! irtn-* 610 6.20 6.30 640 6.50 660 670 660 6.90 700 7.10 770 730 740 7.50 7.60 7.70 7,60 7.90 MVL ......... ........................... ..... ' ........~.......... Amount :________ 0 . 0 0 ~ _____________ i.ion: 131 17.140) ....... ... "" :Resp: 7314 I I I 6.20 6.30 640 6.50 660 6.70 660 6.90 7.00 7.10 7.20 730 7.40 7.50 7.60 7.70 780 7.90 L-d4 Amount: 6.00 ___ ____ m Ion: 135 (7.167) |Reap: 5545 A !\ i\ I\ _6.20_ 630 640 6,50_ 6.60 670 660 690 7.00 7.10 770 730 740 750 760 770 780 7.90 0621018.D PABX BVA.M Wed Oct 18 12:45:33 2000 Page 2 001145 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-90 Page 89 Peak Area Report Sample Kama: 2 xOC MIOWater Notebook Ratarance:PABX-BVA-1-062 DateAcquired: 10/17/2000 1943 Operator: MAN Data FiteName: 0621019.D InstrumentName: MS7 Data FilePath:D:\MS0_DATA\PAB\PA8X-8VAVt-0e2-1\ InstrumentVialNumber: 19 InstrumentMethod Name: PABX_BVA Comoound # 1) j) Compound Name MVL MVL-04 Ratonbon Tane(Min.) !&B 7.14 131 716 135 PeakArea M! 2025 m 4390 Peak Area Ratio 461.3E-3 ST e The rattar **m" in this coturno indicates that manual integration waa pertormed on that compound. Print Oats and Time 10/18/200012:45 Page 1 001146 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-91 Page 90 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BVA\l-062-l\0621019.D Vial: 19 17 0ct~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) itounaencc 200o] 1500 i ioooi TIC:0621019.0 a e, l ii :i ; i i T ' -- |l-1';1 !-- 11^ 'r`V r -. I. ''1 1 ... . I>'. . MTT ' !' Tittx--> 6.10 6.20 6.30 6 *0 6.S0 6.60 6.70 6 J0 6.90 7 00 7.10 7.20 7.30 7.40 7.50 7 60 7 70 7tO 7.90 MVL_________ ~_________________ Amount : ... 0.00______________ I Iol 131 {7.137) iResp: 2025 ~| r-ry-r, 6.20 6.30 640 6.50 660 6.70 6.60 6.90 7.00 7.10 7.20 730 7.40 7.50 760 770 7.60 7.80 (IVL-d4 Amount : .00 Ion : 135 (7.164) ... " Resp: 4390 J \A i\ : 6.20 6.30 640 650 660 6.70 6.60 6.90 7.00 7 10 720 7.30 740 7.50 760 770 780 730 0621019.D PABX_BVA.M Wed Oct 18 12:45:52 2000 Page 2 001147 Primedica-Worcester Project number: PABX-BVA 418-018 :PAGE 1-92 Page 91 Peak Area Report Sampi*Naim:2 xOC LOW Water Notebook Roterence:PABX-8VA-1-062 DataAcquired: 10/17/200020:03 Operator:MAN Data FiteNw n ;0621020.D InstrumentName: M57 DataFilePath:D:\MSD_DATAVPAB\PABX-BVA\1.06MV InstrumentVialNumber: 20 InstrumentMethod Name: PABX_BVA Compound 1) 2) Compound Naim MVL MVL-64 Ratantton Time(Min.) Ion 7.14 131 7.17 13S Peak Area M* 767 m 4380 PeakArea Ratio 175.1E-3 The latter V in th i* column Indicate that manual integration we* parformed on that compound PnntData and Time 10/1B/2000 12:46 Paga } 001148 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\l-062-l\0621020.D Vial: 20 17 Oct 2000 20:03 Operator: MAN 2 x QC l o w water Inst : MS7 MS integration Params: RTEINT.P Q uant M ethod : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE I n te g r a t o r ) ! 2000; l 1500; : 1000; ! 500; TTC: 0621020.D | !1 !] i1 i1 {/ \ Ttnw--> 6.10 6.20 630 6.40 6.50 6.60 6.70 680 6.90 7.00 7.10 720 730 7.40 7.50 7.60 7.70 780 790 MVL Amount: 0.00 i Ion: 131 (7.139) ;.Resp: 767 (\ i /i \ V ! 620 6.30 640 6.50 6.60 670 6.80 6.90 7.00 7.10 7.20 7.30 7.40 750 760 7.70 7.80 730 A flV L -d4 Amount: 0.00 W Ion: 13S (7.167) iResp: 4380 A i t i | 1} 11 j I eio' io . V " j o ' tp'' J o " t a > 7.oo_7io 720 7.30 ;.io 7.50 reo 770 7,o 7.90 0621020.D PABX BVA.M Wed Oct 18 12:46:12 2000 Page 2 001149 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-94 Page 93 Peak Area Report Sample Name: 2xQCILOQ Water Notebook Reference: PABX-BVA-1-062 DataAcquired: 10/17/200020:22 Operator:MAN Data FHeName: 0621021.D InstrumentName: MS7 DataFHe Patti:D:\MSD_DATA\PABlPABX-8VA\1-062-1\ InstrumentVialNumber: 21 instrumentMethod Name: PABX BVA csmwwii 1) 2) Comoound Name MVL MVL-d4 Retention Time (Min.1 Ion 7 14 131 7.17 135 Peak Area M* 361 m 5175 Peek AreaRatio 697.6E-4 M* The letter "m" m m ir colum n indicete th at m ertual integration wa p arto oneri on th at com pound. Pnnt Date and Time 10/18/2000 12:46 Page i 0 0 1 5 0 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-95 Page 94 Quantitation Report (QT Reviewed) Data Pile Acg On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621021.D Vial: 21 17 Oct 2000 20:22 Operator: MAN 2 x QC LLOQ water Inst : MS7 MS Integration Params: RTEINT.P 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 t 01151 Primedica-Worcester Project number: PABX-BVA 418-018.-PAGE 1-96 Page 95 Peak Area Report Sample Name: 21 OC HIGH Placma Notebook Reference:PABX-BVA-1-062 DateAcquimi: 10/17/200020:42 Operator. MAM DateFileName: 0621022.D InurnmentName: MS7 DateFilePath:D:\MSD_DATA\PAB\PABX-BVAV1-002-11 instrumentVialNumber:22 InstrumentMethod Name: PA8X_BVA Comoound * 1) 2) Comoound Name MVL MVL-d4 Retention Tima (Mln.l ISO 7.13 131 7.16 135 PeakAma M* 6581 5208 Peak Atea Ratio 126.4E-2 M* The letter "m" In itile column Indicates that manual Integration was pertOfmad en that compound. Print Date and Time: 10/1B/2000 12:46 l'ge I 001152 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\l-062-l\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: 0621022.0 I\ 2000; 1; I 1000j j{ ' ' 1 ' 1 ` - Tkt -> 6.10 620 6.30 6.40 6 50 6.60 6 70 6 80 6.90 7.00 7.10 7.20 7.30 740 MVL Amount: 0 .0 0 Io n : 131 (7.134) ; R e s p : 6581 A !i \ \ I I 7.60 7.70 7.80 790 ~ ~ I 66..2200 66..:30 640 6.50 6 60 S70 T'l a 0 6.90 7 00 7.1 7 . ' 7.30 ' 7.40 ' 7.50 Amount: 0.00 Ion: 135 T 7 .162) Resp: 5208 A !1 7.70 7.80 7.90 I\ !\ ''T '"1 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 I I 0621022.D PABX-BVA.M Wed Oct 18 12:46:51 2000 Page 2 001153 Primedica-Worcester Project number: PABX-BVA 418-018.-PAGE 1-98 Page 97 Peak Area Report Sam ple Name: 2 x OC MIO Plasma Notebook Refersnee: PABX-BVA-1-062 Dale Acquired: 10/17/2000 21:01 Operator: MAN Data File Name: 0621023.0 Data FUe Path: D:'MSD_DATA\PABVPABX-BVAM -062-1\ Instrum ent Method Nam e: PABX BVA Instrum ent Name: MS7 Instrum ent V ial Num ber: 23 Compound # 1) 2) com pound K atM MVL MVL-64 Retention Ttm tiM iDJ 7.13 7.16 Is a 131 135 PwK A rw C 2280 m 4580 497.6E-3 XT" The M ie -m** Ur Site c o lim i OTdicrteK mat manual integration w et pertermed on mar compound Pnnt Date and Time 10/18/2000 12 47 frage t 001154 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 Parants : RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) 0621023.D PABX_BVA.M Wed Oct 18 12:47:10 2000 Page 2 0 LL55 Primedica-Worcester Project number: PABX-BVA 418-018 :PAGE I-100 Page 99 Peak Area Report Sample Name: 2 x CJC LOW Ptatma Notebook Reference: PABX-BVA-1-062 Date Acquired: 1007/2000 21:21 O perator MAN Data File Name: 0621024.D Data File Path: D:\MSO_DATAVPAB1PABX-BVA\1-062-1\ Instrum ent Method Name: PABX_BVA kietrum ent Name: MS7 Inetnim ent Vial Number: 24 Compound 1) 2) Comnound Name MVL MVL*d4 R etention Tim e fMln.1 7.13 7.16 *911 131 135 Peak Area M* 996 4321 Poafr Ar>4 Rflfr? 230.5E-3 M* The letter "m" in this column indicates that manual Integration was performed on that compound. Print Oate and Time: 10/18/2000 12:47 Page 1 001156 Primedica-Worcester Project number: PABX-BVA 418-018:PA G E 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) Ifcbundai 2000 TIC: 0621024D 1500 1000 500 L _:---- ----------- r-- -- p-" !rinw-> 6 10 6 20 6.30 640 6.50 6.60 MVL j ion: 131 (7.134) !Resp: 996 ; : i--'i-- .., er v r f -- -- p >'. i-- 6.70 680 6.90 7 00 7.10 7.20 7.30 7.40 7.50 Amount: 0.00 r"' n !\ -- i 1 'i1 1 7.60 7,70 7 *0 7.90_. 6.20 630__ 6.40 6 90 6.60 6 70 6.60 6.90 700 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7,80 __-d4 ___ Amount : 0.00 Ion: 135 (7.62T IResp: 4321 A 6.20 6 30 6 4 0 __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 2 4 .D PABX_BVA.M Wed O c t I S 1 2 : 4 7 : 3 0 2 0 0 0 Page 2 O O II5 7 Pnmedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-102 P a g e 101 Peak Area Report Sampta Name: 2 x Control Plasma Notebook R etarne: PABX-BVA-1-062 Dale Acquired: 10/17/2000 21:40 Operator. MAN Data File Name; 0021025.0 Data FMe Patti: D :\M 5D.DAT A'F>AB\PABX-BVA\1.062-1\ Instrum ent Method Nam: PABX_BVA Instrum ent Name: MS7 Instrum ent V ial Num ber. 25 gmawntf i) 2) Compound Nam MVL MVLd4 Retention Tim e (Min.) 7.13 7.16 Isn 131 135 P*kAif oe 525 m 4463 Peek Aree Redo 117.8E-3 M* Th "Wi" tn m l* column indicat* that manual tatesratton wa p fto nw d on ito t cnm potaiii Pont Date and Time. 10/18/2000 12:47 Page 1 00115S Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-103 Page 102 Quantitation Report IQT Reviewed) Data File Acq On Sample Mise 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 Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) 'Abundance 2DOO< 1500-! TIC: 0621Q25D A i. ; 1000* 500 i i . . T- . . Time- 6.10 6.20 6.30 640 6.50 660 6.70 660 6.90 7.00 7.10 7.20 730 740 7.50 760 770 7.60 7.90 MVL Amount: 0.00 Ion 131 (7. Resp 525 t A ! 6.20 6.30 6.40 i bVL-d4 W Ion : 135 (7. 1Resp : 4463 Amount: 0.00 ! 6.20 6.30 6 40 7.10 7 20 7 30 7 40 7 50 7.60 7.70 7 80 730 0621025.D PABX BVA.M Wed Oct 18 12:47:49 2000 Page 2 001159 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-104 Page 103 Peak Area Report Sam ple Nam : 3 x QC HIGH W ats! Notebook Rotatene: PABX-BVA-1-062 Date A cq uim i: 10/17/2000 22:00 Operator. MAN Date FHa Name: 0621026 0 Data File Path: D:\MSD_DATA\PAB1PABX-BVA\1-062-1\ Instrum ent Method Nam : PAflX BVA Instrum ent Name: MS7 Instrum ent Vial Number: 26 Comoound # 1) 2) Compound Name MVl MVL-04 W The tettar *m** Retenbon Tima lMin.1 714 7.16 ISQ 131 135 Peak Area M l 5511 4246 Peak Area Ratio 129.BE-2 wMparfiarmed on that uwnupund Pnnt Data and Time: 10/1B/2000 12:48 Page 1 001160 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-105 Page 104 Quantitation Report (QT Reviewed) Data Pile Acq On Sample M 9 C D:\MSD_DATA\PAB\PABX-BVA\l-062-l\062l026.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) Mundane* 30001 TIC: 0621026.0 I) ii I\ lrT-- > 610 6,20 6.30 6.40 6.50 6.60 670 6.80 6.30 7.00 710 7.20 7 30 7.40 7.50 7.60 7.70 7.60 7.90 MVL Amount: "" 0.00 .... lri: 131 (7.138) Reap: 5511 *\ /V 620 6.30 6.40 6.50 6.60 6 70 6 80 8 90 7.00 7 10 720 7.30 7.40 7.50 7.60 7.70 7,60 7.90 L-d4 __________ ____ Amount: " 0.00 Ion: 135 (7.162) Resp: 4246 I 6.20 6.30 6.40 6.50 6.6 -6.70 6 60 6.90 7 00 7 10 730 7.30 7 40 7.50 7 GO 7.70 7.80 7.90 0621026.D PABX_BVA.M Wed Oct 18 12:48:08 2000 Page 2 011C1 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-106 Page 105 Peak Area Report Sam ple Name: 3 1 QC MIO Water Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/17/2000 2Z19 Operator: MAN Data File Name: 0621027.D Data FHe Path: D:\MSD_DATA\PA8>PA8X-BVA\1-062-1\ instrum ent Method Name: PABX-BVA Instrum ent Name: MS7 Instrum ent V ial Number: 27 Compound 1) 2) ompoun^ Namq MVL MVL-P4 Retention Time (MinJ 7.14 717 |SQ 131 13S Peek Area M" 2059 m 4522 Peak Area Ratio 455.3E-3 M" * 7h* tener *'m" In m u column Indicate mat manual Integration w at patiu tioad en mat compound. P in t Date and Time: 10/18/2000 12:48 Pago J. 001162 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) Abundance TC: 0621027.D ! 2000I ! P !\ 1000) i1 sooi J \ I'1' I ' ' , 1-- r r f-T'- . .. I I I I f 1-- I1 11 1f ' ' ' ' " ' ' I 1 1-- !-- >Tn-> 6 1 0 6,20 6.30 6.0 6.60 6 6 0 6 7 0 6.80 6.90 7.00 7 1 0 7.20 7 30 7.40 7 50 7.60 7 7 0 7.80 MVL Amount: 0.00 790 0621027.D PABX BVA.M Wed Oct 18 12:48:28 2000 Page 2 001163 Priraedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-108 Page 107 Peak Area Report Sample Name: 3 * QC LOW Water Notebook Reference: PABX-BVA.1-062 Date A cquired: 10/17/2000 22:39 Operator: MAN Data File Name: 0621028.0 Data FHa Path: D:\MSD_DATA\PAB\PABX-BVA\1-0e2-1\ Instrum ent Method Name: PABX_BVA bietnim ent Name: MS7 Instrum ent Vial Num ber: 28 Comoound # n 2) Compound Name MVL MVL-d4 Retention Tim e (Min.) 7.14 71B I2Q 131 135 PeeH Area JT 930 m 5310 Peek Area Ratio 175 1E-3 NT * The letter ~m" m this cohann indicate* that manual (mediation eras performed on that compound Print Date and Time 10/18/2000 12 48 Tnqe 1 001164 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-109 Page 108 Quantitation Report (QT Reviewed) Data File : D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621028.D Vial: 28 Acq On : 17 Oct 2000 22:39 Operator: MAN Sample ; 3 x QC LOW Water Inst : MS7 Misc MS Integration Params: RTEINT.P Ouant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) jtoundmee i I 2000i I1 !1 ;1 1000' TIC: 0621028.0 A l\ I fa,,7T-, i-- t .. .... i i -- frne-> 6.10 620 6.30 B4Q MVL i Ion: 131 '(7:1371 iReap: 930 |r r m , |, ,,| -- 6.50 660 6.70 6.80 ..... V - , | ^ ' l l'| i i . i | ' V i | T i l 1; ,> i r i-- , 6.90 7,00 7.10 720 730 740 750 760 7.70 Amount: 0.00 " .... ..... , 11'I 7.80 7.80 i Ion: 135 (7.165) Resp: 5310 !1 -J... 6.20 630 640 6.50 660 670 680 - 6.90 7.00 7.10 720 7.30 7.40 750 7.60 7.70 7.80 780 0621028.D PABX_BVA.M Wed Oct 18 12:48:48 2000 Page 2 001165 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-110 Page 109 Peak Area Report Swnpta N am *: 3 xQ C U .O Q W ater Notebook Raterenee: PABX-BVA-1-0S2 Date Acquired: 10/17/2000 22:58 Oparator: MAM Date Fite Nama: 0021029D Date Ftte Path: D:\MSD_DATA\PAB\PABX-BVAM-062-1\ Instrum ant Method Nama: PABX_BVA htstrum ant N an a: MS7 Inatnim ant Vial Numbar: 29 Compound 1) 2) Comoound Nama MVL M VU 44 Retention Tima (Mhl.) 7 14 7.16 la s 131 135 R ja n A n a t r 353 4789 737.1E-4 U* * Th latter "m" in Oli column indicai (hat manual integration ara partamted on that compound. Pnm Date and Tune. 10/16/2000 12 48 Page J. CO1166 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-111 Page 110 Ouantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621029.D Vial: 29 17 0ct~2000 22:58 Operator: MAN 3 x QC LLOQ Water Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundance 2000i TIC: 0621029.D I/ ;! ,50! 1 i lOOOj i l i ; 5ooI / i.-p-l i '1I!I.I| IIV llT N | .Il"t"j'!! I I1V I , r' .I fTine--> 6.10 020 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 Amount: 0.00 i Ion: 131 (7.137) j Reap: 353 i A i I; i ;\ ___ \ __________________ , 6.20 6.30 6.40 a50 660 670 60 6.90 7.00 7.10 7.20 7.30 740 7.50 7.60 7.70 7.80 7.90 4htVL-d4 Amount: 0.00 W ~ I 5 n : 135 (7.S4) jResp: 4789 \ i\ !\ I : - , i *r ''1 ''T1 I 1......... I16.20 6.30 6 40 6.50 6 60 6.70 6 00 6.90 7.00 7 10 7,20 7 30 7 40 7.50 7 60 7,70 7.80 7.90 0621029.D PABX BVA.M Wed Oct 18 12:49:07 2000 Page 2 001167 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-112 P a g e 111 Peak Area Report Sam ple Nam e: 3 x Q C HIGH Plasma Notebook Reference: PA8X-BVA-1-062 Data Acquired: 10117/2000 2 3 1 8 Operator: MAN Data File Name: 0621030.D Instrum ent Name: MS7 Data FHe Path: O:\MSD_DATA\PAB1PABX-0VA\1-062-11 bistrum ant V ial Number: 30 Instrum ent Method Name: PA8X_8VA C om pound 1) 2) Comoound Name MVL M VL44 Retention ISttieQ SilU 7.13 7.16 Is o 131 135 Peak Area M l 6347 5002 P * Area Ratio 126.9E-2 M* The loner"m " In tni column indicates met menus) intepratlon wee performed en met compound. Print Date and Tune. 10/18/2000 12 49 P/nje 1 0C11G8 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-113 Page 112 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1-062-1\0621030.D Vial: 30 17 OCt 2000 23:18 Operator: MAN 3 x QC HIGH Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method D:\HPCHEM\i\METHODS\PABX_BVA.M (RTE Integrator) ifibunoann 300oi TIC:06210D .. )\ 2000j <I !\ toooj fi I i' I r'f '" i1-- "-r.-T.,.,.i..-- 1I.... Tinw-> 6tO 620 630 6.40 6.50 6.60 670 6.80_ 690 7.(0 710 7.20 730 7.40 750 7.60 7.70 7.80 7.90 M V L .... ............ Amount : 0.00 f lori: 3 (7.134) i Resp: 6347 . I ! fl l\ i i ~T -r 6.20 6.30 6.40 6.50 6.60 6.70 6.90 690,, 7.00 7.10 7.20 7.30 7. 7.50 7.60 7.70 7.80 7.90 -d4 Amount : 0.00 Ion: 135 (7.161) Resp: 5002 A I I 6.20_630.640 ,650 660 670 6.80 6.90 7.00 7.10 7.20 730 740 7.50 7.60 7.70 790 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 (j G1169 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-114 Page 113 Peak Area Report Sample Name: 3 xQC MID Platina Notebook Rafaranca:PABX-BVA-1-062 DateAcquired: 10117/200023.37 Operator:MAN DataF8e Name: 0621031 D InstrumentName: MS7 DataFilePath:D:\MSD_0ATA\PAB\PABX-8VAH-062-1V InstrumentVialNumber.31 InstrumentMethod Name: PABX_BVA Compound 1) 2) Compound Name MVL MVLkM Retention Time IMin.) ton 7.14 131 7.16 135 fakAfP it 2485 4925 PeakArea Rath 504.6E-3 M* The letter "m* in thie eoiieim indicetee met manual intayfation w et performed en that compound. Print Date and Tune: 10/18/2000 12:49 Paqe 001170 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-BVA\l-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\l\MSTHODS\PABX_BVA.M (RTE Integrator) (Mundane* TIC:0621031.D 2000! ;i loooj I / ! '"*'''11 ' ' ',"'I I'1**1-" -"I I '-p-iI'.I ;.. I. frVna-> 610 6.20 6X 6.40 6.50 6.60 6.70 680 690 7JOO 7.10 730 7.30 740 7.90 7.60 7.70 M V L _________________________ Amount :____ 0.00 ! "on: 131 (7.137) I Reap: 2485 60 790 ---- T '"r.... . ''-- 1..... .. 1.. --- - -'*'I'11'I'--Trr-TT"!', 1 6.20 6.30 6.40 650_6.60 6.70 M O 6.90 700 7.10 7.2D 7,30 7.40 7.50 7,60 770 7,60 7 3 0 !71i-d4 Amount: .00 Ion: 135 (7.164) Resp: 4925 i I l* 6.20 630 640 650 660 670 6B0 6.90__700 710 7.20 730 7,40 7,50 7,60 7.70 7.60 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 001171 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-116 P age 115 Peak Area Report Sample Name: 3QC LOW Plasma Notebook Reference:PA8X-BVA-1-062 DafeAcquired: 10/17/200023:57 Operator MAN Oata FileName: 0621032.0 InstrumentName: MS7 Data FilePath:D \MSD_DATA\PAB\PABX-BVA\1-062.1V InstrumentVialNumber: 32 InstrumentMethod Name: PABX_BVA Comoound 1) 2) Comoound Nam MVl MVL-4 Retention Time (Min.) (on 7.13 131 7.16 135 Peak Area M* 661 3850 Peak Area Ratir 223.6E-3 W * Th* lattar Mmn in toit coturni indleaiM that manual intnratton was parlommd on that eunpound. Print Oats and Time: 10/18/2000 12:49 Page 001172 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-117 Page 116 Quantitacion Report (QT Reviewed) Daca File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621032.D T&al: 32 17 0ct~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) Abuidance TIC:0621032.0 A 1500j I 1000i I! i ;1 i 11 500; u rnm*-> 6.10 6.20 6.X 6.40 650 6.60 6.70 6.B0 690 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7. 7.90 MVL Amount : 0.00 Ion: 131 (7.134) i Reap: 861 i\ ! ! I , _ J A _ .. . r*f** I .........., -r-i D6.42V0 630 640 6.50 660 670 6.B0 6.90 7.00 710 7.20 7.30 740 7.50 7.60 770 7.80 7.80 -d4 " Ion: 135 (7.162) Amount: 0.00 Reap: 3850 A T1 6.20 6J__640 6.50 6.60 670 6.80 690 700 710 720 7.30 7.40 7.50 7.60 770 780 7.30 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 001173 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-118 Page 117 Peak Area Report SampiaNam*: 3 1 ControlPlatina Notebook Reference:PABX-8VA-1-062 DateAcqumd: 10/18/20000:16 Operator: MAN DataFiteNama: 0621033.D InatnimantNama: MS7 DateFitePath:D 1MSD_DATA\PAB1PABX-8VA\1-062-1! InxtrumantVialNumber: 33 teatrumentMethod Nama: PABX_BVA Compound 1) 2) Comoound Name MVL MVL-44 Retention Tima (Min.) Ion 7.14 131 7.16 135 P- Area J 561 m 4846 Peak AreaRatio 119.9E-3 n r a Tha tonar V In m is colum n inoicatna that m anual m tagrapon waa partaim ad on that compound. Pnnt Date and rime: 10/18/2000 12:50 rarjt* 1 001174 Primedica-Worcestcr 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\l-062-l\0621033.D Vial: 33 18 Oct 2000 00:16 Operator: MAN 3 x Control Plasma Inat : MS7 MS Integration Parana: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC:0621033.0 f: 2000; \\ i ;isooij i! I: iooo; I i /\ sooi Tn*-> 6.10 6.20 6. 640 6.30 660 670 6B0 6.90 7.00 7.10 720 7.30 740 7.S0 7.60 7.70 7.60 790 MVL Amount: 0.00 ! Ion: 131 (7.137) :Reap: 581 Ii Ii ii 620 630 6.40 6.50 6.60 6.70 680 6.90 7.00 7.10 7.20 730 7.40 7.50 7.60 7.70 760 7.30 L-d4 Amount': 0 . 0 0 ____________ 9 F . Ion: 135 (7.161) ! Reap: 4846 A !\ !\ I 620 6.30 6.40 6.50 6.60 6.70 660 690 700 7.10 720 730 7.40 7.50 7.60 7.70 7.60 7.90 0 6 2 1 0 3 3 . D PABX_BVA.M Wed O c t 18 1 2 : 5 0 : 2 4 2000 Page 2 001175 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-120 Page 119 Peak Area Report Sample Name: 4 QC HIGH Watw Notebook Referanee:PABX-8VA-1-06? DateAcquired: 10/18120000.38 Operator:MAN DataFHe Name: 0621034.0 InetrumentName: MS7 DataFHe Path:O:\MS0_DATA\PABVrABX-SVAM-062-1\ InetrumentVialNumber: 34 InetrumentMethod Name: PABX_BVA Comoound # 2) Comoound Name MVL MVL-d4 Retention Time IMin.) tan 7.14 131 716 135 PeakArea M* 4620 3568 Peak AreaRatio 129.5E-2 M* * T b t tetter NmMm th is colum n in d icate* th at m anual integration w as pertori--d on th at ecm pcuiui. Print Date and Time: 10/16/2000 12:50 Page 1 001176 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\l-062-l\0621034.D 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\l\METHODS\PABX_BVA.M (RTE Integrator) Abuntenc ! 2500 2000 TiCr0621034.0 A. isooj looo; soo- " 1' 610 6.20 6.30 6.40 6S0 660 670 6.80 6.90 7.00 7.10 720 7.X 7.40 750 7.60 770 7.80 750 MVL Amount : 0.00 i lori: 131 (7.13 9) !Resp: 4620 JnI II \ \ \\ \t '. 1I.I1' -- 11. '11I1'1 I'1; "I........... .'iTTV'-riTi>T'-'-.-,r' ' -ri-rr-- *"(->-- r-t-r-ry 6.20 6.30 640 650 6.60 6.70 6.80 690 7.00__7.10 720 7.30 7.40 7.60 7.60 7.70 7.60 790 ^jjV L-d4 Amount :_______ 0 . 0 0 ______________________ W Io n : 135 ( 7 . 6 2 ) ............................................. Resp: 3568 !i i\ 6.20 6.30 640 660 660 670 680 690 700 7.10 7.20 7.30 740 7.50 760 770 780 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 001177 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-122 P ag e 121 Peak Area Report Sampla Nam: 4xQC MIOWatar Notebook Reference:PABX-BVA-1-062 DateAcquimi: 10/180000ftSS Operator:MAN Data FileName: 0621035.0 InstrumentName: MS7 DataFHa Path:D:\MSO_DATA\PABVABX-BVA\1-062-1\ InstrumentVialNumber: 35 InstrumentMethod Name: PABX_BVA Comnound f 1) 2) Comnound Name MVL MVL-4 Retention Time/Mln.l tea 7.13 131 7.16 135 Peak Area M* 2193 m 4777 Peak Ama Rabe 459.1E-3 M* The M tar V in this n iu m n Iw ciiw tfwt menue/ iw n y itwn w m perte w w d on O ut compound. Pont Date and Tima: 10/18/2000 12:50 *<}* 1 001178 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\l-062-l\0621035.D Vial: 35 18 OCt 2000 00:55 Operator: MAN 4 x QC MID Water Inst : MS7 MS Integration Parama: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abundara 2500- 2000 1SOO TIC:0621036.0 h ii /i 1000 \ i500; ' -I-.-" f r " 'i " " I I i'T-. I " . .......p fTirm-> 6.10 6.20 6.30 6.40 6.50 6.60 670 6.80 6.90 7.00 710 7,20 7.X 740 7JO MVL Amount : Ion: 131 (7.133) Resp: 2193 760 770 7.80 790 ft !\ - /V 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 i 7.00 7.10 i > 7.20 7.X 7.- 7.50 7.60 7.70 780 7.90 MVL-d4 Amount : 0.00 " Ion: 135 (7.161) Resp: 4777 O o 0 1 I i 1 6.20 6.30 6.40 6.50 660 6.70 6.80 6.90 7.00 7.10 7.20 730 7.40 750 7.B0 7.70 780 790 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 001179 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-124 P age 123 Peak Area Report Sample Name: 4xOC LOW Water Notebook Reference:PA8X-8VA-1-062 DateAcquired: 10/16/2000 115 Operator.MAN DataFileName: 0621036.D InctnimentName: MS7 Data FilePath:D:\MSD_DATA\PAB\PABX-BVA\1-062-1\ InstrumentVialNumber:36 InstrumentMethod Name: PABX_BVA Comoound# 1) 2) Comoound Name MVL MVL44 Retention TimtfMin.] 1SQ 7.14 131 7.16 135 PaekArea M* 835 4968 PanArea Ratio 168.1E-3 M* a Tit* tatter "m" in thia eotumn M i a t H that manual Integration < m paitcirmad on that compound. Print Date and Time: 10/18/2000 12:51 Page 1 00I 1 0 Primedica-Worceter Project number: PABX-BVA 418-018:PAGE 1-125 Page 124 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\1- 062 -1\0621036 .D Vial : 36 18 0ct~2000 1:15 Operator: MAN 4 x QC LOW water Inst MS7 MS Integration Parants: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) S kb trid rc ! -I ! 20! !; i is ii " TIC: 621096.D a !' n ;I ! ....!' ---!' " -- I.III'> |l /\ ,-- IV III-- .it;fi.,,!.1.,II,.IIII,IIi.II,1-r,I trim- 6.10 6.20 6.30 6.40 650 6. 6.7P 6 80 680 7. 710 7.20 7.30 7.40 7.50 7.60 770 7 7.90 MVL Amount: ." 0 6 2 1 0 3 6 . D PABX_BVA.M Wed O c t 18 1 2 : 5 1 : 2 2 2000 Page 2 001181 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-126 Page 125 Peak Area Report Sample Name: 4 xOC LLOQ Water Notebook Reference: PABX-BVA-1-062 DateAcquired: tortB/20001:34 Operator.MAN DateFHeName: 0621037.0 ImtninwntName: MS7 DataFilePath: D:\MSD DATA\PAB\PABX-BVA\1-062-1\ tnetnimentV W Number: 37 InetrumentMethod Name: PABX_BVA Comoound# 1) 2) Compound Name MVL MVL-4 Retention Time(Min.) iSB 7.14 131 7.16 135 PeekAree M* 365 5012 Peek AreaRetio 72B.3E-4 M* The lattar *m* in thl* column indicate that manual integratten waa peflomwd on that compound. Pnnt Date ana Time: 10/18/2000 12:51 Page 1 001162 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-127 Page 126 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSDJDATA\PAB\PABX-BVA\l-062-l\0621037.D Vial: 37 18 0ct~2000 1:34 Operator: MAN 4 x QC DLOQ Water Inst : MS7 MS Integration Params: RTBINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) Mund*no> j 25001 20001 '! 15001 j i Uj TC:0621037.0 j I I i I j r "*" 11I1" I I1" I 1 - r r , II-,. .. .. III.II,I.I,1II . I.... Tim- 6.10 6.20 630 6.40 6.50 8.60 6.70 6.B0 6.90 7.00 7.10 730 7.30 740 7.50 7.00 7.70 7 . K 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 001163 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-128 Page 127 Peak Area Report SampleName:4 *OC HIGH Plasma Notebook Reference:PASX-8VA-1-062 DateAcquired:1On8/2000 1:54 OperatorMAN DataFReName: 0621038.D Inabunwnt Name: MS7 DataFilePath:D:\MS0 DATAV>AB\PABX-SVA\1-062-1\ InstrumentVialNumber.38 InstrumentMethod Name: PA8X_BVA Compound 1) 2) Compound Name MVL MVL*d4 Retention Time (Mln.l lac 7.13 131 7.16 136 Reek Aree 1 6P37 790 Peek AreeRatio 1316E-2 i r Th tetter *mMin ttwa colum n indicate that m anual tm agratton me pertorm ed m that compound. Pnnt Date and Tim: 10/18/2000 12:51 Paq* 1 001164 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\1-062-1\0621038.D Vial: 38 18 Oct--2000 1:54 Operator: MAN 4 x QC HIGH Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTK Integrator) itaundm I SOOOi OOo! i! 3000 T ic: 0621036.0 A i\ !I Ii 2000! ! 1000! 1I I- J"TTM' 1 1 1 I 1 ' i ............. I . , . r r . V u' 1 , . . . . n T . . . . . . . . . . . ................... ,. fTim*-* 6.10 620 6,30 6.40 6.SD 6.60 6.70 6.60 620 7.00 7.10 7.20 7 . 7.40 7.S0 7.60 7.70 760 7.90 MVL Amount: 0.00 I Ion: 131 (7.133) !Resp: 8937 i\ i\ iI 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6 90 7.00 710 7.20 7.30 ' 7.60 7 5 0 .....760 7,70__ 7.80 "7.90.' 7L-d4 Amount : 0.00 Ion: 135 (7.161) Resp: 6790*6 6 20 6 30 640 i' 6 50 ................- 6.60 6 70 6.60 r-- 1 1 I . I i I . I I . I I . I I ! I . r I I I I . I , . . . ,T r n .T ,., . ... . , 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 8 . D PABX BVA.M Wed O c t 18 1 2 : 5 2 : 0 1 2 0 0 0 Page 2 0011S 5 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-130 Page 129 Peak Area Report Sample Nam: 4 x OC MIO Plasma Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/16/2000 2:13 Operator: MAN Data File Name: 0621039.0 Data FBe Rath: D:\M5D_DATA\PA8\PAaX-BVA\1-062-1\ Instrum ent Method Name: PABA_BVA Instrum ent Name: MS7 Instrum ent V ial Number. 39 P w w af 1) 2) Compound Name MVL MVL-4 Retention Tlm eJM ln.) 7.13 7.16 iSQ 131 135 PoahArea M ! 2156 4477 F M K V ta Ratto 481.6E-3 M* * The lettor Tn* In thl coturni! Ind icf a met monna! tntogrotten woe pe rformed on met compound. Pnnt Dale and Time: 10/18/2000 12:52 Page 1 S60 0 1 1 Primedica-Worcester Project number: PABX-BVA 418-018:PA G E 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: HAN 4 x QC MID Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) taundanoe" I; i 2000 TIC: 0621039.D 1500 ioooj i 500,i ; ; I .Tim- 6.10 6.20 MVL 6.30 6 40 p.r.i', 650 6 60 r,- T T I T . | - | | 1 '^ . I i - f r i . I . I , f i -1 I I I I--------f , l- T ' H ' 1 " 6 70 6.60 650 7.00 7.10 7.20 7.30 7.40 .7.60 _T 6p JT.70 . 7.60 Amount: 0 . 0 0... . 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 O O lls ? Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-132 Page 131 Peak Area Report Sam ple Nh m : 4 x QC LOW Ptam a Notebook Rit m n a : PABX-BVA-1-062 M e Acquired: 10/160000 2:33 Operator: MAM Data FU* Maine: 0621040.D Data File Path : D:\MSD_DATASPABiPABX-BVAV1-062-11 instrum ent Method Name: PABX_BVA Inalnim ant Name: MS7 bmbumant Vial Number: 40 Comoovnd # 1) 2) Compound Name MVL MVL-d4 Retention Time IMInJ 7.1 7.16 Is a 131 135 PaakAm a 1156 5064 Peak A n a R at 227.4E-3 NT- The letter "m " to th u cotumn indteetem that mam aI tmagrattofi was nerlonwad an that compound P iin t Data and Time: 10/18/2000 12:52 Page 1 001X88 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 U W Plasma Inst : MS7 MS Integration Pararne: RTE1NT.P Quant Method : D:\HPCHEM\l\MBTHODS\PABX_BVA.M (RTE Integrator) TIC: 0621040.0 20001I f. ,1500 1000 i I! i; h 500 ' ! r r r -- -- I . " . . , r '- - ? I r --' I1 1 r -- ` : ' 1 -- -- > Ime- 6.10 6.20 6.30 6.40 6JO 6.60 6.70 S.ao S J0 7.00 7.10 7 20 7.30 740 750 7.60 7.70 7.60 7.90 MVL Amount : 0.00 I Ion: 131 (7.137) !Reap: 1156 fi I\ 6 2 0 6.30 6 40 6.S0 6.60 6.70 6.60 6 3 0 TOO 7.10 7 3 0 7.30 7.40 7 SO 7.60 7.70 7.80 7.90 s L -d4 ion: 135 (7.161) Amount: .... 0.00 .... ....... Reap: 5084 \ /\ / \. '-- 17- 620 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 0 6 2 1 0 4 0 . D PABX BVA .M Wed Occ 1 8 1 2 : 5 2 : 4 0 2 0 0 0 Page 2 1101109 Primedica-Worcester Project number. PABX-BVA 418-018.-PAGE 1-134 Page 133 Peak Area Report S w n pit Nam: 4 x Control P in n a Notabook Reference: PABX-8VA-1-C62 Date Acquired: 10118/2000 2 5 2 Oparator: MAN D ata File N ana: 0821041.0 Date FIte Path: O:\MSO_OATA\PAB\PABX-BVAM-062-1\ Instrum ent Method Name: PABX_BVA Instrum ent N an a: MS7 Instrum ant V ial Num bar. 41 Com aound 1) 2) Compound Name MVL MVL-d4 Retention Tima (Mln.1 7.13 7.16 &Q 131 135 PsslAwa M* SBS m 4680 Paak Area Ratio 1 25 4 E -3 M* * Tbs lattsc V in this column Indicates that manual M sgraticn ass aadoffnad on that compound. Pont Date and Time: 10/18/2000 12:52 Pape 1 001190 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 0ct~2000 2:52 Operator: MAN 4 x Control Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHBM\l\METHODS\PABX_BVA.M (RTB Integrator) ^Dundanea i 2000 TIC: 0631041 .D fi 1500 \1000 500 I Tgw--> 6.10 6-2D 6.30 640 6.50 8-60 6.70 6.80 6.80 7.00 7.10 7.20 7.30 7.40 750 7 60 770 7 80 7.90 MVL_______________ Amount: 0.00 j Ion: 131 (7.133) :Reap: 588 !\ i 6.2D 6 . 6.40 6.50 6 60 6.70 6.B0 6 90 7.00 7 10 7.20 730 740 7.50 760 7.70 7.W 7.90 -d4 Amount: 0.00 ion: 135 (7.160) Resp: 4690 !\ ; ! 6-20 6.30 640 6.50 660 6.70 6JO 6JO 7 00 7 10 7-20 7.30 7 40 7 50 ~7S0 770 7 . 7.90 I i 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 U1131 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-136 Page 135 Peak Area Report Sample Naina: 5xQCHIGHWalar : PABX-BVA-1-062 Date Acquami: 10/18/2000 312 Operator: MAN Da File Name: 0621042.0 testtumantName: MS7 Data FilePatti: D:\MSO_DATA1PAB\PABX-BVA\1-062-1\ InmtnjmentVtel Number 42 tesbument Method Name: PABX_BVA Comooundt 1) 2) 9HP9MMI Nani MVL MVL-d4 Retention Tim tffllQ J 7.14 7.16 (2D 131 135 EaMats M: 7137 5531 PtnDAWDJWo 12S.0E-2 M* * Tho latter V In this column tndleatea (hot manual Injapration waa partormart on mat Prml Date end Time HV1&2000 12:53 Pge 1 001192 Prnedica-Worccster Project number: PABX-BVA 4 1 8 -0 1 8.-PAGE 1-137 Page 136 Quantitation Report (QT Reviewed) Data File Acq On Sangle Mise D:\MSD DATA\PAB\PABX-BVA\l-062-l\0621042.D Vial: 42 18 0et~2000 3:12 Operator: MAN 5 x QC HIGH Water Inst : MS7 MS Integration Pararas: RTEINT.P Quant Method : D:\HPCHEm \i \m e t h o d S\p a b x _BVA.M (RTE Integrator) TIC:0621042.D ; 3000ji h i i 2000j 10; ji i ; -- ... i" -.... -i -rr- 'I 1 I'-'''`rT ' - r r . .-,-n irn~ 6.10 620 630 6.40 6 50 660 670 6.60 6.90 7.00 7.10 ... 7J20 7D 7.40 7.S0 7.60 7 70 7,80 7.90 MVL Amount: .od Ion: 131 (7.137) Resp: 7137 i l -I I1' .. I t ..>; .,Tp-,-r^-rr- [ 620 6 3 0 6 4 0 6.50 6.60 6.70 6.60 6 9 0 7.00 7.10 720 7,30 740 730 7.60 770 7.60 7.90 ' ' '-d4..... Ion: 135 (7.164) jResp: 5531 A m ount:________ 0 . 0 0 rt H j _____________ L V _ ......................... ' I 1--1 r ! n : ' 1 . ' i . 1 1I ' ' ! ................ .! --------1 ' I ' ' " .............. .. ' 1 ' I 6.23... 6 3 0 6.40 6.50 6 .60 6.70 6.60 6.90 7 00 7.10__ 7.20 7 3 0 7 40 7.S0 7 60 7 70 7.80 7 90 0 6 2 1 0 4 2 . D PABX BVA.M Wed O c t 1 8 1 2 : 5 3 : 1 8 2 0 0 0 Page 2 001193 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-138 Page 137 Peak Area Report Samp* Name: 5 OC MIO W ater Notebook R o ta rn e *: PABX-8VA-1-062 D a ti A cq uim i: 1(V I8/2000 3:31 Operator: MAN Data Fila N aim : 0621043.D Oata FAa Path: 0^MSD_DATAV>AB\PASX-eVA\1-062-1\ Instrum ant Method Name: PABX_BVA kutrum ant N am *: MS7 Inrtrum ant V ial Num ber. 43 Compound* 1) 2) Compound Nam* MVL MVL-4 Retention TimtIMinJ Ian 7.13 131 7.16 135 Em KA ff fiC 2277 5024 Peak Area Ratio 4532E-3 M*The1titor"m"inthitcolumninaieMnPiaiimmiti intaerallenm> ptrfonntaonbut compound. Print Date and Time: 10/18/2000 12:53 Pqe i 001194 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-139 Page 138 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621043.D Vial: 43 18 Oct 2000 3:31 Operator: MAN 5 x QC MID Water Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) TIC: 06MCM3.D 2000! /j tOCOi I I\ 1V 'T^'r 111 1I" " 1 .. !111 I1 111 1I1 1I'l'''II I' !1I 11*I :.. l- !rm - 6.10 6 .20 6.30 640 6.S0 6.60 6.70 6.B0 S3C TOO 7.10 7-20 7 J0 7 M 7 X J .6 0 770 7,30 .700. MVL ' Amount: . Ion: 131 (7.134) !Resp: 2277 !\ \ i 620 6.30 ' .4 0 6SO 660 670 6.80 630 7.00 7.10 700 7.30 7 . 7.50 730 7.70 7.K I ' 70 -d4 Amount: 0.00 W ion: 135 (7.161) iResp: 5024 .... r >-. , , r r-T ; - , - . I , ..... T , , . . - ; r r rrp-, 620 6.30 9.40 6 50 6.60 670 6.80 6.90 7.00 710 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 0621043.D PABX_BVA.M Wed Oct 18 12:53:38 2000 Page 2 001195 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-140 Page 139 Peak Area Report Sem pt* Nam: 5 * Q C LOW Wetar Notebook Rotatene*: PABX-BVA-1-082 Dote Acquired: 10rt6/2000 3:50 Operator: MAN Date FU* Nomo: 00210*4.0 Dote File Poto: D:\MSD_DATAV>ABV>ABX<6VA\1-062-1\ Inetnim ent Mothod Norm : PASX_BVA tectounont Noma: MS7 H ntnim ent Vini Number: 44 C om pound* 1) 2) Comoound Nome MVL MVL-44 Rete n tion T h ne/M ln .1 7.13 7.16 22 131 135 P e e kA im w r 685 4124 Peek Ateo Rat 168.5E-3 M* Tiw tettpc *m ' Ht mi coturno Hrdte f e thet manuel integratio n w n performed on ttie t mmpeund. Print Date end Time. 10/16/2000 12:53 Pag 001196 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-141 Page 140 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise 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 Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Abmdancc 2000 TIC: 0621044 0 1500 iooo! j soo! ! v- -T - , I 1 1 1 6.10 630 630 6.40 650 MVL Ion: 131 (7.134) Resp: 695 1 IT T - ' ' * | " ' 60 670 6.60 6.80 '-`T .:. I ' 1 " '1 ............ * T T ' n - T l -- - r - i f . T P - I TOO 7.10 7.20 7.30 740 7SO 760 7 70 7-60 7.60 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 0G1197 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-142 Page 141 Peak Area Report Sample Name: 5 x QC U.OQ Water Notebook Reference: PA8X-BVA-1-0e2 Data Acquired: 10/18/2000 4:10 Operator: MAN Data File Name: 0021045.0 Inatnimant Name: MS7 Data File Patti: D:\MSO_DATA\PABVPABX-BVA\1-062-1V bntnim ant Vial Number: 45 Instrument Method Name: PABX_BVA Comoound f 1) 2) Compound Nama MVL MVL-04 Retention Tima (Mbi.) 7.14 7.16 Ks 131 135 PeakArea M* 314 4172 Peak Aree Ratio 752.0E-4 M*The latter "m" leethi column wcwm that marniti integration partocmefl w that compound. Print Date and Time: 10/18/2000 12:54 Page 1 0C1138 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\1\METH0DS\PABX_BVA.M (RTE Integrator) tobuntfanc* i 20001 TIC: 0621045.D I I 1500j I loooj H 1i l! I sooi iI TI**> rr-, iti m II ..i.< n .rJHwi=f'-,J-rT\rrm |.. .......... .... . ffim- 6.10 620 6.30 660 6.60 6.60 670 660 6.90 7.00 7.10 7,20 7.30 7.40 7.50 7.60 7.70 7,60 MVL _______________ Amount: _ 0 . 0 0 ______ ; h: 131 (7.138) I Resp: 314 '1 7.90 i si 6.20 6.30 6.40 6.50 6.60 6 7 0 6.60 ' 6 9 0 " 7.00 7. ' 7.20 7.30 7.40 7 6 0 7,60 7 70 7.60 7.90 -d4 Amount:.. .0 Ion: 135 (7.162) ;Resp: 4172 ft i\ I !\ ! 620 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 7.70 7 80 7.90 0 6 2 1 0 4 5 .D PABX_BVA.M Wed O c t 10 1 2 : 5 4 : 1 7 2 0 0 0 Page 2 0G11S9 Primedica-Worcester Project number. PABX-BVA 418-018:PAGE 1-144 Page 143 Peak Area Report Sam ple Name: 5 * OC HK3H Ptaema Notebook Reference: PABX-BVA-1-062 D ate Acquired: 10/18/20004:28 O perator MAN Data File Name: 0621046.0 Data File Path: D:\MS0_DATA\PA8\PABX-eVA\1-062-1\ instrum ent Method Name: PA8X_BVA Inatnim ent Name: MS7 Instrum ent V ial Num ber. 48 Com pound 1) 2) Compound Narna MVL MVL-4 R etention S a n . (M h l) 7.13 7.16 Jsn 131 135 EssSAiw t t 7218 5671 PeaXAiea Ratio 127.3E-2 M* The leuer "m" In mia column Indicate s that manual i n t e g r a t i o n wm4 purtormed on that rnupinsal Pnnt Date and Tim: 10/18/200012:54 Peg* 1 o c 1200 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 Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\MBTHODS\PABX_BVA.M (RTE Integrator) ffjciundanc I 4000; TIC:0621M6.0 300o| ;I ! ;I i 2000 1000; Tim- 8.10 6.20. 6.30 6.40 680 660 670 6.60 6.90 7,00 7.10 7,20 7. 7.40 7.90 7.60 7.70 7.60 790 MVL ________ Amount : 0.00 ! 'ion: 131 (7.133) i Resp: 7218 ft I/\ nn 1I ' I 1' 1 : 1' ' 1, ' ' T--T I .-.-i-l I I . '<I I I I I 6.20 6.30 640 6.50 660 6.70 6.60 6.90 700 7.10 7,20 7.30 7.40 750 7.60 770 7.80 7.80 -d4 Amount : .00 Ion: 135 (7.160) Resp: 5671 M i 6.20 6.30__640 6.50 6.60 670 6.60 690 70 7.10 7.20 7.30 740 7.50 7.60 770 760 7.60 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 001201 Primedica-Worccster Project number. PABX-BVA 418-018.PAGE 1-146 Page 145 Peak Area Report Sample Hw m : S x O C M tt ) Ptaema Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/16/2000 4:49 Operator: MAN Data File Name: 0621047.0 Data FDa Patti: D:\MSO_DATA\PAB1PABX-BVA\1.062-1\ Inatrum ent Method Name: PABX_BVA Inebum ent Name: MS7 In a tn jifw ll Vial Number: 47 Comoound 1 U 2) Comoound Ni im MVL MVL-d4 Retention Tim a (Min.1 7.14 7.16 bo 131 135 P ff." .A ^ t c 2502 5132 Poak Ama R ate 487.5E-3 M* The letter "m" in m il column indiealaa that manual mtagrahan m peffom ad an that compound. Pnrrt Date and Time: 10/16/2000 12:54 Page 1 001202 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-147 Page 146 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621047.D Vial: 47 18 Oct 2000 4:49 Operator: MAN 5 x 0C MID Plasma Inst : MS7 MS Integration Parame: RTEINT.P Quant Method : D:\HPCHBM\l\METHODS\PABX_BVA.M (RTE Integrator) fOEdT ii i; 200o! 10001 TIC: 0621047.0 A j: !\ i- I`1:'1''I " 1 p-.-p. i11,M I11 rm-> 610 6.20 630 6*0 6.50 6.B0 6-70 6.60 6.90 7 00 7.10 7J20 7.30 7.40 7,50 7.60 770 780 7.90 MVL______________ ________ __ Amount: 0.00 . Ion: 131 (7.136) i Resp: 2502 I I 6.20 6 30 6.40 6.50 6.60 6 70 6.80 690 7.M 710 7 a i ;io-nd: 135 4 (7.160).. _______________ Amount: Reap: 5132 A i*. 7 . 7 40 ' 7.60 7 60 7 p 760 730 0.00 ........................ .1 ' ' lj1I ... r' ' ..11(11111! i; i.>Ii(<IiI 1-.;. 6.20 6.30 6 40 6 50 6.60 6 70 660 6JO 7,00 7.10 7 20 7 . 7.40 7 50 7 60 7 70 7.60 7.90 0621047.D PABX BVA.M Wed O c t 18 1 2 : 5 4 : 5 5 2 0 0 0 Page 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-148 Page 147 Peak Area Report Sample Name: S x QC LOW Plasma Notebook (M etan os: PABX-BVA-1-062 Dale Acquired: 10/18/2000 S OS O p erator MAN Data FHa Name: 0 6 2 1 0 4 )0 Data FHe Path: D:\MSD_0ATA 'f'AB\PABX-BVA\1-082-n Instrum ent Method Name: PABX_BVA m stnim ant ama: MS7 Inatrum ant V ial N um ber 48 Compound 1) 2) NT The lattar Compound Name MVL M VL44 Retention Tim elM in.l 7.13 7.16 la c 131 135 e a ilL & B C 744 3350 Peak Area Ratio 222.1E-3 va nerftitead ca that compound. Print Oats and Tima: 10/18/2000 12:55 Page 1 001204 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\062104B.D Vial: 48 18 Oct 2000 5:08 Operator: MAN 5 x oc LOW Plasma Inst : MS7 MS Integration Pararne : RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M IRTE Integrator) pw8no i ISOOj TIC:0621048.D 1000! S00| yv-vyri-r-." ;'rl -T'-,y .1.r. r , .TTr ! p Time-> 6.10 6.20 6.30 6.0 6.50 6.60 6.70_6.80 6.90. MVL " ........ .~ | Ion: 131 (7.134)... .... I Resp: 744 ;- | - .~T'y'-. . i . | i'l- iT M 7.00 7.10 7.20 7.30 7 *0 Amount:____0.0_0_ ...................... T- . i ................................. ' ' 7.50 7.60 7 70 7 80 730 ___ ______ I\ \ '"'1-- -r-,----I- r- ....... ..p -- .ir-f " I ...I ' I 6.20 6.30 6,40 6.60 6.60 670 6.80 6.90 7.00 7.10 7.20 7,30 7.60 760 7.60 7.70 7.80 7.90 -d4 Amount : 0.00 Ion: 135 (7.161) Resp: 3350 IV i 620 6.! 6 40 6.S0 6.60 6 70 6.80 69o"~ 7.00 710 7 20 7.30 7.40 7.50 760 7.70 780 790 062104B.D PABX_BVA.M Wed Oct 18 12:55:14 2000 Page 2 001205 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-150 Page 149 Peak Area Report Sam ple Name: 5 > Contrai R aim a Notebook Reference: PA8X-BVA-1-062 Data Acquired: 10/180000 5:28 Operator: MAN Data File Name: 062104B.D Data File Patti: D:\MSD_DATA\PA8\PABX-eVAU -062-1\ Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Instrum ent Vial Num ber 49 Com pound* 1) 2) Compound Name MVL MVL-d4 R etention Tim e (Mm.) 7 .1 3 7.16 tea 131 135 EsHflrw K . 306 3344 Peak Area Retto 118 4E-3 t jr The ietlsr "m" in M ill ceiumn indici e l thsi marnili iniegretien m i portoimod on that con d a n n i. Pont Date and Tima: 10/18/2000 12:55 Prtqe 1 001206 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\l-062-l\0621049.D Vial: 9 18 Oct 2000 5:28 Operator: MAN 5 x Control Plasma Inst : MS7 MS Integration Params: RTBINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M IRTE Integrator) AbundantI I ! 15001 jI ! 1000 TIC: 0621048.0 H ;I I r . - - :T W T C r-r-> M ' /\ .. / \ - .irpw .,. , , , . , ,.-,1 , |-p" . - | ..................-"P **....... ! ~ 6.10 620 6.30 6.40 6.80 6.60 t.70 6.90 6.90 7.00 7.10 7.20 7 X 7.40 7SD 7.60 7.70 7.80 7.90 m v l Amount: ' 0. 0 6 2 1 0 4 9 . D PABX BVA.M Wed O c t 18 1 2 : 5 5 : 3 4 2 0 0 0 Page 2 001207 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-152 P a g e 151 Peak Area Report Sample M u m : 6 x QC HIGH Water Notebook Reference: PABX-BVA-1-082 Date Acquired: 10/18/2000 5:47 O perator MAN Otoe FUe Name: 0021050 0 Otoa FUe Path: D:\M5D_DATA\PABIPABX-8VAtf-062-11 Inetrum ent Method Name: PABX_BVA tnetrum enl Name: MS7 Intoniment V ial Num ber. 50 C w nw w rty i) 2) Compound Name MVL MVL-44 M" The M a r "m* R d m tio fl Tune (M in.) 7.14 7.17 tan 131 135 Peek Aree W 7184 5458 Peak Aree Ratio 131.6E-2 Me pertermad on that compound. Print Date end Time: 10/18/2000 12:55 Pacte 1 001208 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-153 Page 152 Quantitation Report (QT Reviewed) Data File Acq On Sam ple Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621050.D Vial: 50 18 Oct 2000 5:47 Operator: MAN 6 x QC HIGH Water Inet : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHQi\l\METHODS\PABX_BVA.M (RTE Integrator) kbunamoa ! 4000l TIC: 0621060.0 3000-; i| ! 2000' Ii1 ii ! 1000! i\ r* !Twm~ 10 6.20 'I '-T '1T * * '1l' 6.30 6.40 650 6.60 6.70 6.80 6.90 _t1 7.0 710 V 720 ''720 7.40 750 7.60 7.70 7.00 7.90 MVL Amount 0.00 i Ion: 13 P?.138) !Reap: 7184 j ft l\ j 6.20 630 6.40 6.S0 6.60 6.70 f!r r-r 6.00 6.90 7.00 1 ! \V. 7.10 7.20 7.30 7.40 750 7.60 7.70 7.00 i 760 kVL-d4 Amount : 0.00 W Ion: 135 (7.166) !Reap: 5458 | i i I i 6.20 6.30 6.40 650 6.60 6.70 6.60 6.90 ft l\ 700 7.10 7.20 7 .X 7.40 7.50 7.60 7.70 780 7.90 11 0 6 2 1 0 5 0 . D PABX BVA.M Wed O c t 18 1 2 : 5 5 : 5 3 2 0 0 0 Page 2 001209 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-154 Page 153 Peak Area Report Sam ple Name: 6 xQC MID Watar Notebook R ehrenea: PAHX-8VA-1-062 Data Acquired: 10/18/2000 6:07 Operator: MAN Data Fila Mama: 0621051.D D ata Fila Patti: D:\MSD_DATA1PAB\PABX-BVA\1-062-1\ Inabum ant Mathod Name: PABX_BVA Inatniment Name: MS7 Inatnim ant V ial N um ber 51 Retention Compound Mama Tim a (M in.) tea Peak Area M* Peak Ama Ratio 1) MVL 7.13 131 2956 m 468.5E-3 2) M V L-d t 7.16 135 6314 KT Tke M tM V knthit ce*um* in tealM that manuel IntognMen me partorvned an that comp* Pnnt Data and Time: 10/1R/2000 12:56 Page I 0C1210 Primedica-Worcester Project number: PABX-BVA 418-018-.PAGE 1-155 Page 154 Quantitation Report (QT Reviewed) Data File Acq On Semple 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 Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) fcbuntone* 1 t;` XDOl 11 TIC: 0621051.D A i j 2000j i\ \ 1000 ii i\ !\ 610 620 6.30 6.40 6.50 60 6.70 680 6.90 700 7.10 7.20 7 30 7 . 780 7 60 7 70 7.80 780 MVL Amount: 0.00 1 ion: 131 (7.134) Resp: 2958 6.20 6.30 6.40 6 50 6.60 670 680 EOO 7.00 7.10 7.20 7,30 740 750 7.60 770 7 80 7.90 -d4 """ Amount: 0.00 Ion: 135.(7.162) Resp: 6314 A, \\ I v. 1 l|,` ,1 .1"1'''1I'1 - '*1' 111II "'T1' !II,II1I1 >I' 1J', '111 620 6 .X 6.40. JSS0 _ 6.60 670 6.60 6.90 7.00 7.10 720 7 .X 7.40 7.S0 760 ! 1''T-T--T-T-r 7 70 7 80 7.90 0 6 2 1 0 5 1 .D PABX_BVA.M Wed O c t 18 1 2 : 5 6 : 1 2 2 0 0 0 Page 2 001211 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-156 P age 155 Peak Area Report Sam ple Name: 6 x QC LOW W ater Notebook Reference: PABX-8VA-1-062 Data Acquired: 1On8/2000 8:26 Operator: MAN Date Pile Name: 0621052.D Data R ie Patti: D:\MSD_DATAVPAB\PABX-BVA\1-062-1\ Instrum ent Method Name: PABX-BVA Irwkum ent Name: MS7 Inetnim ent V ial Number: 52 Comookind # 1) 2) Comoound Name MVL MVL-da Retantton Tlm e/M ln.) 7.13 7.16 te a 131 135 E fte kA m 1039 6187 ifa k .A ffii.B ta p 1C7.9E-3 M*s The letter Tn" in this u tlwmn indtcelee that manuel integration wee pertermeU en Stai cemeeund. Print Dale am) Time: 10/18/2000 12:56 Pepe 1 001212 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-157 Page 156 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise 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 Parana : RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) Adunitene ! ""I TIC: 06210S2.D ! 2000! !j Ii ! 1000! jrm-> 6.10 620 6.30 6.40 MVL Ion: 131 (7.135) Resp: 1039 6 . r to' 6.W l|1\ ) 6' I 't.w ' T m T.20~ 7.M 7.40 7.50 7.60 ~ T 70 7.80 T _______Amount:______ 0.00_________________ .I M /\ . I J. >. I ; . , I I . I I . , I I ) I I 11' f PTTPfrffr>'.-lT >-r. - ft . f 6.20 6.30 6-40 6.50 6.60 6.70 6.B0 6.90 7 0 0 7.10 ___7_3_D___ 7.30 7 40 7.80 7 60 7.70 7.60 7.90 d4 ~ ________ _ _ _ Amount: 0.00 r,Ion: 135 (7.159) IResp: 6187 A ! 6.20 6 3 0 6.40 6 6 0 6.60 6 7 0 6 8 0 6.90 7.00 7 1 0 7.20 7.30 T.40 7.50 7,60 7 7 0 7.80 7.90 0 6 2 1 0 5 2 . D PABX BVA.M Wed O c t 1 8 1 2 : 5 6 : 3 2 2 0 0 0 Page 2 001213 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-158 Page 157 Peak Area Report Sample N aim : 6 x QC 0 .0 0 W ater Notebook Ratetanca: PABX-BVA,14362 D ateA equirad: 10/18/20006:46 Operator: MAN Data File Name: 0621053.D data Fite Pate: D:\M SD_0A7A\PABV>ABX-aVA\1-082-H Instrum ent Method Name: PABX_BVA Inamm en! ama: MS7 teatrw nant Vial Num ber: 53 Compound 11 2) Compound Nemo MVL MVL-d4 R etention Tim e IM in.l 7.14 7.16 ten 131 135 Peek Area M" 462 m 6048 Peak A rta Rabo 763.06-4 IT > The tette **mMIn itile column Indicete* that menuet integration wee performed on that compound. Print Dole end Time: 10/18/2000 12:56 Page 1 001214 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\l-062-l\0621053.D Vial: 53 18 Oct"2000 6:46 Operator: MAN 6 x QC LLOQ Water Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHBM\1\MBTH0DS\PABX_BVA.M (RTE Integrator) MVL ion: 131 (7.135T Resp: 462 Amount: 0.00 i\ l\ T-- >-!- 'r^i-1-1'11I11I">1I H '<>I.-.-rI'r-- 630 6.30 640 6 .50.....6.60 6.70 6.BO 6.90 7.00 710 7 . 7 . 7.40 "I-'-' 7.60 760 7. 7JI0' 'JI'1 790...__ L-d4 ... Amount : ... 0 -00... ......~~ ion: 135 "('7.163) ......... "TM iResp: 6048 ^ -r 630 6 .X 6.40 6.50 6.60 6.70.... 6.80 690 7.00 710 720 7.30 7 40 7.50 7.60 7.70 7.60 7.90 i 0 6 2 1 0 5 3 . D PABX BVA.M Wed O c t 18 1 2 : 5 6 : 5 1 2 0 0 0 Page 2 001215 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-160 Page 159 Peak Area Report Sam ple Name: 6 x QC HIGH Plasma Notebook Reference: PABX-BVA-1-062 Data Acquired: 10/18/2000 7:05 Operator: MAN Data File Nam e: 0621054.0 Data FHe Path: D:\MSO_DATA\PAB\PABX-BVA\1-062-U Instrum ent M ethod Name: PABX_BVA Instrum ent Name: MS7 taatnim ant V W Number: 54 Comoound # 1) 2) Comoound Name MVL MVL-4 Retention Tlm elM ln.) 7.13 7.16 IBS 131 135 Peak Area M l 4406 3506 Peak Area Rebe 125.6E-2 ITaThe lattar"m" inthisautumnindicatesthatmanual imegfulianareaperform ad anthatcreipound. Pnnt Date and Time: 10/18/2000 12.57 rag* i 001216 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-161 Page 160 Quantitation Report (QT Reviewed) Data Pile Acq On Sainple Mise D:\MSD DATA\PAB\PABX-BVA\l-062-l\0621054.D Vial: 54 18 Oct- 2000 7:05 Operator: MAM 6 x QC HIGH Plasma Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) MMMftt 22550001J TK: 0621064.!0\ 2000; isoo; i 1000 900 / ^ r T -.T -- -n - . - n - T - . - T T f n n - 1 I 6.10 K70 6 30 6 40 6.50 6.80 870 680 690 T OO 7.10 7JO 7JO 7.40 7 50 7.60 7.70 7.00 7.90 MVL Amount: 0.00 ________________ Ion: 131 (7.132) Resp: 4406 A iI _L-5L6.20 6.30 6.40 6 .50 6 60 6.70 6 80 3 0 7 00 7.10 7JO 7.30 7.40 7 6 0 7.60 7.70 7 j o ' 7 3 0 L-d4 Amount: 0.00 Ion: 135 (7.159) !Resp: 3508 iA I /V T-- 620 .6.30 6 40 6.50 6.60 6.70 6.60 6.90 7 00 7.10 7JO 7.30 7.40 7.50 7.60 7.70 7.60 790 0621054.D PABX BVA.M Wed Oct 18 12:57:10 2000 Page 2 OC1217 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-162 Page 161 Peak Area Report SampleName: BxQC MIOPlaama Notebook Reference:PABX-8VA-1-062 DateAcquired: 10/18/20007:25 Operator:MAN Date FHaName: 0821055.D bwtninianlNam*: MS7 DateFN*Path:O:\MSD_DATA\PAB\PA8X-BVA\1-082-1\ InatnimantVialNumber: 55 InetrumentMethod Name: PABX_BVA Compound# 1) 2) pmraw Nww MVL MVL-d4 Retention Time(Min.) tan 7.13 131 7.18 135 PeakAtea M* 2452 4988 PeakArea Rat 4B1.6E-3 M* T l lanar "m" in t h ii colum n indicata th a m anual integration wan parlormad an Oiat com pound. Pnnt Dale and Time: 10/18/200012:57 Page 1 001218 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-163 Page 162 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621055.D Vial: 55 18 Oct 2000 7:25 Operator: MAM 6 x QC MID Plasma Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) pbraiane TIC:0621065 0 I 2000' 1000! !I \ !\ ^ 'T ! ' I ' >~l T'-'V' 'W | .< T-- Tme-> 6.10 6-20 6.30 6.40 650 6.60 670 680 690 7.00 7.10 7.20 7.30 7.40 7.50 7,60 7.70 760 7.90 MVL .. '_ Amount:______ 0. ______ j Ion: 131 (7.133) ! Resp: 2452 iI n 1 I... I I"!'T-H t *....... . 8.20 6. 6 40 6.50 6.60 6.70 6.60 6.90 TOO 7.10 7.20 730 7.40 7.80 7.60 7.70 7.80 7.90 d4 Ion: 135 (7.161) Amount: 0.0 I ! Resp: 4988 A \ 1 7-r-pr-.-r .-pri - ^ - ,'T 'I I 6.20 6 30 6.40 6.50 6.60 670. 6 60 690 700 7.10 7.20 7 30 7.40 7.50 7.80 7.70 7.80 7.90 0621055.D PABX BVA.M Wed Oct 18 12:57:30 2000 Page 2 001219 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-164 Page 163 Peak Area Report Sample Mama: 6 itOC LOW Platina Notebook Reference:PABX-BVA-1-062 DateAcquired: 10/18/20007:44 Operator:MAN DateFHe Name: 0621056.D InstrumentName: MS7 DateFilePath:0:\MSO_DATA\PABVPABX-BVAM-062-1\ InstrumentV W Number: SS InstrumentMethod Name: PABXBVA Compound 1) 2) Compound Name MVL MVL-4 Retention TimefMln.l Ion 7.14 131 7.16 135 Pnhfttf BE 1176 5136 PeakArea Rstlo 226.9E-3 ITThelottar"n r inthi*columnmehcri-- dialmanualtntograttonwas porforvnatfontrial compound. Punt Date and Time: 10/18/2000 12:57 Page 1 001220 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\PAfl\PABX-BVA\i-OS2-X\0621056.D Vial: S6 18 0ct~2000 7:44 Operator: MAN 6 x QC LOW Plasma Inst : MS7 MS Integration Pararne: RTEINT.P Quant Method : D:\HPCHEM\l\M8TH0DS\PABX_BVA.M (RTE Integrator) 0621056.D PABX BVA.M Wed Oct 18 12:57:49 2000 Page 2 001221 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-166 Page 165 Peak Area Report Sample Name:6 xControlPlasma Not*book Rafsranca:PABX-BVA-1-062 DaleAcquired: 10/18/20008:04 Operator MAN DataFKaName: 0621057.D InstrumentName: MS7 DataFIlaPatti:D:\MS0 DATA\PABIPABX-BVAM-062-1\ InttnimenlVialNumber 57 toetrumantMethod Name: PABX_BVA Comoound 0 1) 2) Compound Name MVL MVt-d4 TimeIMIn.l ten PeakArea M* PeakArasRatio 7.13 131 503 115.0E-3 7.16 135 373 MTThaIettar"m"InmilcolumnmdteelMthmanualtnlegreUonm * ptrtprmiflonIW oiiyuund. Print Data andTtma: 10/18/2000 12:58 Page 1 001222 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-BB A PRIMEDICA ARGUS 905 Sheehy Drive, Building A H orsham , PA 19044 001223 418-018:PAGE 1-187 O P rim edica 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 H orsham , PA 19044 Submitted by: Primedica-W orcester 57 Union Street Worcester, MA 01608 Primedica Report # PABX-BBA-01-145 Page 1 o f 187 Issue Date: November 7, 2001 VJ-.Out ttfrh rk A. Netsch / Date Project Scientist Bioanalytical Chemistry Department Primedi ca-W orcester 001224 G P rtmfdtc.a Primedica-Worcestcr 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 o f good laboratory practice. Official Journal o f 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 001225 O P rimrdtca 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 o f QAU audit/inspections for GAS CHROMATOGRAPHIC-MASS SPECTROM ETRIC ANALYSIS OF MEVALONIC ACID IN EDTA PLASMA FO R ARGUS STUDY 418-018, Project Number PABX-BBA. Critical Phases 1.Laboratory Procedure 2. Raw Data 3. Draft Final Report D ate Inspected 01/23/2000 03/29/2001 04/19/2001 D ate R eport Subm itted to Study D irector M anagem ent 04/27/2001 01/29/2001 04/27/2001 04/03/2001 04/27/2001 04/26/2001 The Final Report GAS C H RO M A TO G RAPH IC-M ASS SPEC TR O M E TR IC ANALYSIS OF M EVALONIC ACID IN EDTA PLASMA FO R 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 o f good laboratory practice. Official Journal o f 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 Aui Date 001226 GPRTMFDTfA 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 STATEM ENT............................................................................... 3 LIST OF TABLES AND APPENDICES................................................................................. 5 CONTRIBUTING PERSONNEL............................................................................................. 6 ANALYTICAL STANDARD CHARACTERIZATION/STABILITY................................7 ARCHIVAL STO RA G E............................................................................................................ 8 SUMMARY.................................................................................................................................. 9 Sample R eceipt............................................................................................................... 9 Method Summary............................................................................................................ 9 Calibration Standard Accuracy and Precision.............................................................9 Quality Control Sample Accuracy and Precision..................................................... 10 Study Sample Concentrations...................................................................................... 10 CONCLUSIONS.........................................................................................................................10 001227 G P r im fo ic a Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAG E 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 o f Interpolated Mevalonic Acid Lactone QC Standard Concentrations............................................................................................................... 13 TABLE 3 Summary o f Mevalonic Acid Lactone Least-Squares Linear Regression Constants and Analysis Dates...................................................................................... 14 TABLE 4 Summary o f Mevalonic Acid Lactone Concentrations in EDTA Rat Plasma Study Samples.................................................................................................. 15 APPENDIX A Representative Chromatographic Data - Batch No. PABX-BBA-1-047-1............ 23 001228 GPRTMF.DF.A Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-192 Page 6 Primedica Argus Study 418-018 CONTRIBUTING PERSONNEL Project Scientist.................................................................... Director, Analytical Chemistry............................................ Analytical Chemist................................................................ ...........Yanira Villalobos, A.C.E. Report Coordinator............................................................... 001223 GPRTMF.DICA 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 o f the Supplier, as is the method o f synthesis, fabrication or derivation and stability determination. 001230 O P rtmfdtca 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 o f 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-BB A -01-145. 001231 GPRTA4F.nO 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 o f December 5, 2000 and February 13, 2001. The samples were stored at <-20C until the time o f 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 o f 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 o f mevalonic acid lactone from 100 pL EDTA rat plasma samples over a concentration range o f 10.0 ng/mL to 250 ng/mL. The complete description o f the assay and summaries o f 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 o f each batch. A copy o f representative raw chromatographic data from the first batch o f 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 o f samples in an analytical run. For each analytical run, a l/X 2-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 o f 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 GC1232 OPRTMEnrfA 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 o f 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 o f 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 o f 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 lim it o f 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 o f 50). Interpolated concentrations for all remaining samples are expressed in ng/mL. Refer to Table 4 for concentrations o f mevalonic acid lactone in rat plasma study samples. CONCLUSIONS: All analytical results were within acceptable limits with the exception o f one value reported as an AQL estimate due to insufficient sample available for reanalysis. 001Z 3 3 Primedica-Worcester Project Number PABX-BBA 418-018:PAGE 1-197 Page 11 TABLES 001234 Prnedica-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 PA B X -B B A -1-054-1 PA B X -B B A -1-056-1 PA B X -B B A -1-058-1 PA B X -B B A -1-062-1 PA BX-BBA-1-065-1 PA B X -B B A -1-067-1 PA B X -B B A -1-069-1 PA B X -B B A -1-072-1 PA B X -B B A -1-075-1 1M 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 20.0 29.6 NR 48.6 48.9 94.9 91.6 152 150 19.8 19.3 19.8 20.0 21.3 20.6 30.3 30.1 30.9 30.2 30.7 30.9 48.4 51.0 49.0 51.2 48.1 48.1 88.4 91.5 87.8 89.4 89.4 87.0 150 155 150 150 152 152 20.2 20.5 30.5 31.0 48.6 50.0 89.2 91.2 153 151 19.6 18.6 19.4 22.9 19.0 20.6 30.5 30.9 30.2 31.4 29.6 32.4 46.7 47.6 46.8 53.1 52.2 49.6 85.6 93.8 87.0 92.0 90.8 89.6 149 163 148 152 155 147 19.7 30.4 48.5 92.1 N R N R 51.3 92.0 144 159 N R 34.1 46.1 91.0 19.1 31.0 45.5 88.6 145 163 250 249 245 246 251 248 254 249 248 242 244 255 255 235 245 239 240 239 246 249 251 M ean Std. Dev. n %RSD % N om inal 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 001235 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-199 Page 13 TABLE 2 SUMMARY OF INTERPOLATED MEVALONIC ACID LACTONE QC STANDARD CONCENTRATIONS Batch ID Theoretical Nominal Concentrations (ng/mL) 25.0 70.0 200 P A B X -B B A -1-047-1 22.1 53.6* 74.3 67.2 199 184 P A B X -B B A -1-054-1 23.5 24.9 65.4 70.0 183 187 P A B X -B B A -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 P A B X -B B A -1-062-1 23.0 24.1 64.2 65.5 187 188 P A B X -B B A -1-065-1 PABX-BBA-1-067-1 PABX -BBA -1-069-1 P A B X -B B A -1-072-1 21.9 26.5 22.4 28.6 23.5 23.2 21.1 23.2 64.1 64.0 66.8 68.4 66.4 63.1 69.1 69.8 184 192 183 182 187 188 200 192 PABX -BBA -1-075-1 29.3 23.9 61.6 62.5 184 179 M ean Std. Dev. n %RSD % N om inal 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 G rubbs' Test. Value excluded from summary statistics 00123G 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 PA B X -B B A -1-047-1 Slone 0.0067118 PA B X -B B A -1-054-1 0.0062552 PA B X -B B A -1-056-1 0.0061528 PA BX -BBA -1-058-1 0.0060042 PA BX-BBA-1-062-1 PA BX -BBA -1-065-1 PA B X -B B A -1-067-1 0.0061041 0.0064579 0.0064673 PA B X -B B A -1-069-1 0.0063214 PA BX-BBA-1-072-1 PA B X -B B A -1-075-1 0.0064268 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 -0.01975 0.99919 0.99652 02/13/01 02/15/01 001237 Primedica-Worcestcr Project Number: PABX-BBA 418-018:PA G E 1-201 Page 15 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal # -Groun #-Generation Concentration fne/m Ll Batch ED 10902-G R O U P 1-F0 10903-GROUP1-F0 10904-G R O U P 1-F0 10905-GROUP1-F0 10906-G R O U P 1-F0 10907-G R O U P 1-FO 10908-G R O U P 1-FO 10909-G R O U P 1-FO 10910-G R O U P 1-FO 10911-G R O U P 1-FO 10912-G RO U P 1-FO 10913-G RO U P 1-FO 10914-GROUP 1-F0 17.3 23.7 26.3 26.0 36.0 205 36.5 32.1 20.7 35.1 19.8 34.2 23.3 PA BX -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 PABX-BBA-1-054-1 10901-G R O U P 1-FI 10902-G R O U P 1-FI 10903-G R O U P 1-F 1 10904-G R O U P 1-F 1 10905-G R O U P 1-FI 10906-G R O U P 1-F 1 10907-G R O U P 1-FI 10908-G R O U P 1-F 1 10909-GROUP1-F1 10910-G R O U P 1-F 1 10911-G R O U P 1-F I 10913-G R O U P 1-F 1 37.9 36.8 BQL (DF = 10, <100 ng/mL) 40.8 44.3 BQL (DF = 10, <100 ng/m L) 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-FO 10919-GROUP2-FO 10920-G R O U P2-F0 10921-GROUP2-FO 10922-G R O U P2-F0 10923-G R O U P2-F0 10924-G R O U P2-F0 10925-G R O U P2-F0 10927-G R O U P2-F0 10928-G R O U P2-F0 2300 1550 1130 1500 1850 1140 99.7 522 159 432 1780 218 2060 PABX -BBA -1-058-1 PABX -BBA -1-058-1 PABX -BBA -1-058-1 PA B X -B B A -1-058-1 PA B X -B B A -1-058-1 PA B X -B B A -1-058-1 PABX-BBA-1-054-1 PABX -BBA -1-058-1 PABX-BBA-1-054-1 PABX-BBA-1-058-1 PABX -BBA -1-058-1 PABX-BBA-1-054-1 PABX-BBA-1-058-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 001238 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 Animal # -G rouo #-G eneration Concentration (n e Jm L ) Batch ID 10915-G R O U P 2-F 1 10916-G R O U P 2-F 1 10917-GROUP2-F1 10918-GROUP2-F1 10919-G R O U P 2-F 1 10920-G R O U P 2-F 1 10921-G R O U P 2-F 1 10922-GROUP2-F1 10924-G R O U P 2-F 1 10925-G R O U P 2-F 1 10927-G R O U P 2-F 1 10928-GROUP2-F1 29300 AQL (estimated 23,200) 15100 17000 24000 12900 671 168 326 17400 821 17100 PA B X -B B A -1-058-1 P A B X -B B A -1-058-1 P A B X -B B A -1-062-1 P A B X -B B A -1-062-1 PABX-BBA-1-058-1 P A B X -B B A -1-062-1 PABX-BBA-1-058-1 PA B X -B B A -1-054-1 P A B X -B B A -1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PA B X -B B A -1-062-1 10929-GROUP3-F0 10930-G R O U P3-F0 10931-GROUP3-F0 10932-GROUP3-F0 10933-G R O U P3-F0 10934-G R O U P3-F0 10935-GROUP3-F0 10937-GROUP3-F0 10938-GROUP3-F0 10940-G R O U P3-F0 10941-GROUP3-FO 10942-G R O U P3-F0 158 1670 1440 1220 1440 866 1530 1410 451 1380 416 327 P A B X -B B A -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 PA B X -B B A -1-062-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 10929-GROUP3-F1 10 930-G R O U P 3-F 1 10931-GROUP3 -F 1 10932-GROUP3-F1 10933-GROUP3-F1 10934-G R O U P 3-F 1 1093 5-GROUP3-F1 10937-GROUP3-F1 10940-G R O U P 3-F 1 10942-G R O U P 3-F 1 156 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 BQ L = Below quantitation limit. A Q L = Above quantitation limit DF = Dilution Factor * = Estim ated value, DF = > 12,500 ng/mL 001239 Prixnedica-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 A n im a l # -Grout) #-Generation Concentration fne/mLl Batch ID 10943-G R O U P4-F0 10944-G R O U P4-F0 10945-G R O U P4-F0 10946-G R O U P4-F0 10947-G R O U P4-F0 10948-G R O U P4-F0 10949-G R O U P4-F0 10950-G R O U P4-F0 10951-GROUP4-FO 10952-GROUP4-F0 10953-G R O U P4-F0 10954-G R O U P4-F0 10955-G R O U P4-F0 10956-G R O U P4-F0 133 172 1590 2460 1600 161 760 1730 1560 99.1 1160 1000 203 24000 P A B X -B B A -1-047-1 P A B X -B B A -1-047-1 P A B X -B B A -1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-047-1 P A B X -B B A -1-058-1 PABX-BBA-1-058-1 PABX-BBA-1-058-1 P A B X -B B A -1-047-1 P A B X -B B A -1-058-1 P A B X -B B A -1-058-1 P A B X -B B A -1-047-1 P A B X -B B A -1-058-1 10945-G R O U P 4-F 1 10946-G R O U P 4-F 1 10947-G R O U P 4-F 1 10949-GROUP4-F1 10950-G R O U P 4-F 1 10951-G R O U P 4-F 1 10 953-G R O U P 4-F 1 10 954-G R O U P 4-F 1 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-G R O U P 11-FO 11031 -GROUP 11-F0 11032-G R O U P 11-FO 11033-G R O U P 11-FO 11034-G R O U P 11-FO 11035-GROUP11-FO 1103 5-G R O U P 11-FO 11037-G R O U P 11-FO 1103 8-G R O U P 11-FO 61.4 59.7 22.0 27.5 797 37.3 48.0 38.0 42.6 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-058-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 11029-GROUP 11-FI 11031-GROUP11-F1 11033-G RO U P 11-FI 1103 6-GROUP 11-FI 11037-GROUP 11-FI 17.4 11.4 20.2 17.8 15.7 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 PABX-BBA-1-047-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 001240 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 A nim al # -Groun #-Generation Concentration (ne/mL) Batch ID 11040-GROUP12-F0 11041-G R O U P 12-FO 11042-G R O U P 12-FO 11044-G RO U P 12-FO 11045-GROUP12-F0 11046-G RO U P 12-FO 11047-G R O U P 12-FO 11048-GROUP12-F0 11049-G R O U P 12-FO 11050-GROUP12-F0 11051-G R O U P 12-FO 11052-G R O U P 12-FO 15.5 24.8 22.3 31.5 15.5 18.9 22.5 BQ L (DF = 1, <10 ng/m L) 14.0 14.9 24.2 21.9 P A B X -B B A -1-056-1 PA BX -BBA -1-056-1 PA BX -BBA -1-056-1 PA BX -BBA -1-056-1 PABX-BBA-1 -056-1 PA BX -BBA -1-056-1 PA B X -B B A -1-056-1 P A B X -B B A -1-056-1 PA B X -B B A -1-056-1 PA B X -B B A -1-056-1 PA BX -BBA -1-056-1 PA B X -B B A -1-056-1 11040-GROUP12-F1 11044-G R O U P 12 -F 1 1 1045-G R O U P12-F1 11046-G R O U P 12-F 1 11047-GROUP12-F1 11050-G R O U P 12-F 1 11051 -G R O U P 12-F 1 11052-G R O U P 12-F1 52.5 54.4 31.6 190 87.3 33.7 55.6 31.0 PA B X -B B A -1-056-1 PA B X -B B A -1-056-1 PA B X -B B A -1-056-1 PA BX -BBA -1-056-1 PA B X -B B A -1-056-1 PA B X -B B A -1-056-1 PA BX -BBA -1-056-1 P A B X -B B A -1-056-1 11053-GROUP 13-F0 11054-G R O U P 13-FO 11055-GROUP 13-F0 1 1056-G R O U P13-F0 11057-GROUP 13-F0 11058-G R O U P 13-FO 11059-GROUP 13-F0 1 1060-G R O U P13-F0 11061-G R O U P 13-FO 11062-G R O U P 13-FO 11063-G R O U P 13-FO 11064-G R O U P 13-FO 11065-G R O U P 13-FO 11066-G R O U P 13-FO 28.3 524 B Q L (DF = 1, <10 ng/m L) 28.8 20.8 29.4 36.7 15.0 19.2 23.8 22.9 18.0 66.9 17.3 PA BX -BBA -1-056-1 PA BX -BBA -1-062-1 PA B X -B B A -1-056-1 PA BX -BBA -1-056-1 PA BX -BBA -1-056-1 PA B X -B B A -1-056-1 PA BX -BBA -1-056-1 PA BX -BBA -1-056-1 P A B X -B B A -1-056-1 PABX-BBA-1-056-1 PA B X -B B A -1-056-1 P A B X -B B A -1-056-1 PABX-BBA-1-056-1 PABX-BBA-1-056-1 11053-G RO U P 13-F1 1 1054-G RO U P 13-F1 11056-G R O U P 13-F1 1 1058-G R O U P13-F1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 32.0 199 41.3 64.0 P A B X -B B A -1-056-1 P A B X -B B A -1-056-1 P A B X -B B A -1-056-1 P A B X -B B A -1-056-1 0C1241 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 # -Grout) #-Generation Concentration Cne/mLl Batch ID 11059-GROUP 13-F1 11060-GROUP13-F1 11062-G R O U P 13-F1 11063-GROUP13-F1 11066-G R O U P 13 -F 1 BQL (DF = 2, <20 ng/mL) 185 71.1 194 51.9 P A B X -B B A -1-056-1 PA B X -B B A -1-056-1 P A B X -B B A -1-056-1 PABX -BBA -1-056-1 PABX -BBA -1-056-1 11501-G RO U P 1-FO 11502-GROUP 1-FO 11503-GROUP1-F0 11504-GROUP1-F0 11505-G RO U P 1-F0 11506-G RO U P 1-FO 11507-G RO U P 1-F0 11508-GROUP 1-FO 11509-GROUP 1-FO 11510-GROUP 1-FO 11511-GROUP 1-FO 1 1512-GROUP1-FO 11513-GROUP1-FO 11514-G RO U P 1-FO 27.1 30.2 26.9 BQL (DF - 2, <20 ng/mL) 107 22.8 17.8 25.5 19.0 18.9 28.0 27.2 20.8 26.4 PA B X -B B A -1-065-1 PA B X -B B A -1-065-1 P A B X -B B A -1-065-1 PA B X -B B A -1-065-1 PA BX-BBA-1-065-1 PA B X -B B A -1-065-1 PA BX -BBA -1-065-1 PA B X -B B A -1-065-1 P A B X -B B A -1-065-1 PA BX -BBA -1-065-1 P A B X -B B A -l-065-1 P A B X -B B A -1 -065-1 P A B X -B B A -l-065-1 P A B X -B B A -l-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 1 1524-GROUP2-FO 1 1525-GROUP2-FO 11526-GROUP2-FO 11527-GROUP2-FO 11528-GROUP2-FO 173 161 92.8 160 135 413 135 190 32.7 146 89.6 356 111 P A B X -B B A -l-0 6 5 -1 P A B X -B B A -l-065-1 P A B X -B B A -l-0 6 5 -1 P A B X -B B A -l-0 6 5 -1 P A B X -B B A -l-0 6 5 -1 PA B X -B B A -1-072-1 P A B X -B B A -l-065-1 P A B X -B B A -l-065-1 P A B X -B B A -l-0 6 5 -1 P A B X -B B A -l-0 6 5 -1 P A B X -B B A -l-065-1 PA B X -B B A -1-072-1 P A B X -B B A -l-065-1 11529-GROUP3-FO 115 3 0 -G R O U P 3 -F 0 11532-GROUP3-FO 11533-GROUP3-FO 126 429 1180 821 PABX -BBA -1-067-1 PA B X -B B A -1-072-1 PABX -BBA -1-072-1 PA B X -B B A -1-072-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 001242 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 # -G rouD #-Generation Concentration fne/m D Batch ID 11534-GROUP3-FO 1 1535-G RO U P3-FO 11536-GROUP3-FO 11537-GROUP3-FO 11538-GROUP3-FO 11539-GROUP3-FO 11540-GROUP3-F0 11541-GROUP3-FO 11542-GROUP3-FO 11543-GROUP3-FO 128 8210 375 141 411 2530 173 3680 311 347 PA BX -BBA -1-067-1 P A B X -B B A -1-072-1 P A B X -B B A -1-072-1 PA BX-BBA-1-067-1 P A B X -B B A -1-072-1 P A B X -B B A -1-072-1 PABX-BBA-1-067-1 PA BX -BBA -1-072-1 PA BX -BBA -1-072-1 P A B X -B B A -1-072-1 11544-GROUP4-FO 11545-GROUP4-FO 11546-GROUP4-FO 11547-GROUP4-FO 11549-GROUP4-FO 11550 -G R O U P 4 -F 0 11551-GROUP4-FO 1 1552-G RO U P4-FO 11553-GROUP4-FO 11554-GROUP4-FO 1 1555-G RO U P4-FO 11556-GROUP4-FO 11557-GROUP4-FO 6970 1550 9720 164 82.1 494 483 457 308000 ** 389 648 320 541 PA BX -BBA -1-072-1 PA BX -BBA -1-072-1 PABX-BBA-1-075-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 P A B X -B B A -1-072-1 P A B X -B B A -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-GROUP 11-F0 11632-GROUP11-FO 11633-G R O U P 11-F0 11634-GROUP 11-F0 11635-G R O U P 1 1-FO 11636-G R O U P 1 1-FO 11637-G R O U P 11-FO 11638-GROUP11-FO 11639-GROUP 11-F0 48.1 25.4 25.8 29.6 80.8 40.3 38.7 35.8 38.3 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor ** = V alue was confirm ed 001243 Primedica-Worcester Project Number PABX-BBA 418-018:PAGE 1-207 Page 21 TABLE4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal # -Grouo #-Generation Concentration (ne/mL'l Batch ID 11640-G R O U P 12-FO 11641-G R O U P 12-FO 11642-G R O U P 12-FO 11643-GROUP12-FO 11644-G R O U P 12-FO 11645-G R O U P 12-FO 11646-G R O U P 12-FO 11647-G R O U P 12-FO 11648-G R O U P 12-FO 11649-G R O U P 12-FO 11650-GROUP12-F0 11651-G R O U P 12-FO 11653-GROUP12-FO 41.3 34.2 47.6 14.6 25.8 17.5 26.5 35.5 102 19.7 27.3 24.6 38.1 P A B X -B B A -1-067-1 PABX-BBA-1-067-1 PA B X -B B A -1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PA B X -B B A -1-072-1 PABX-BBA-1-067-1 P A B X -B B A -1-067-1 11654-G R O U P 13-FO 11655-G R O U P 13-FO 11656-G R O U P 13-FO 11657-GROUP13-FO 11658-GROUP13-FO 1 1659-G R O U P 13-FO 11660-GROUP13-F0 11661-G R O U P 13-FO 11662-GROUP13-FO 11663-G R O U P 13-FO 11664-G R O U P 13-FO 11665-G R O U P 13-FO 1 1667-G RO U P13-FO 32.4 30.5 39.5 11.9 11.8 B Q L (D F = 1, <10 ng/m L) 38.1 31.2 16.8 42.0 28.0 B Q L (D F = 1, <10 ng/m L) BQL (D F = 1 , <10 ng/mL) P A B X -B B A -1-069-1 PABX-BBA-1-069-1 PA BX -BBA -1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -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 1 1501/11506-G R O U P 1-F 1 11504-G R O U P 1-FI 11502/11507-G R O U P 1-FI 11505-G R O U P 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-G R O U P 1-FI 11509/11510-GROUP1-F1 11512/11511-GROUP1-F1 11513/11514-GROUP1-FI 26.4 25.8 23.3 20.5 PABX-BBA-1-069-1 PABX-BBA-1-069-1 PABX-BBA-1-069-1 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 PA B X -B B A -1-072-1 PA B X -B B A -1-072-1 001244 Primedica-Worcester Project Number: PABX-BBA 418-018.PAG E 1-208 Page 22 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES A n im al # -Groun #-Generation Concentration Cne/mLl Batch ID 11519/11518-GROUP2-F1 11521/11516-GROUP2-F1 11523/11527-GROUP2-F1 11525/11528-GROUP2-F1 11526-GROUP2-F1 275 239 214 152 49.0 P A B X -B B A -1-072-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -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 PA B X -B B A -1-072-1 193 PA BX -BBA -1-069-1 243 PA B X -B B A -1-069-1 340 PABX-BBA-1-072-1 11547-GROUP4-F1 11552-GROUP4-F1 11630/11633-GROUP11-F1 11632/11636-GROUP 11-FI 11635/11637-GROUP 11-FI 11638-GROUP11-F1 11639/11634-G R O U P 11 -FI 51.3 339 21.7 24.0 BQL (DF = 2, <20 ng/mL) BQL (DF = 2, <20 ng/mL) 21.4 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 P A B X -B B A -1-069-1 11640/11641-G R O U P 12 -F 1 11643/11647-G R O U P 12-F 1 11646/11644-G R O U P 12-F 1 11648/11651 -G R O U P 12-F 1 11650-G R O U P 12-F 1 11653/11642-G R O U P 12-F 1 23.3 BQL (DF = 4, <40 ng/mL) 14.7 21.9 22.4 BQL (DF = 2, <20 ng/mL) P A B X -B B A -1-069-1 P A B X -B B A -1-072-1 PA B X -B B A -1-069-1 P A B X -B B A -1-069-1 PA BX -BBA -1-072-1 P A B X -B B A -1-069-1 11654/11662-G R O U P 13-F1 11656/11655-GROUP 13-F1 BQL (DF = 2, <20 ng/mL) BQL (DF = 2, <20 ng/mL) P A B X -B B A -1 -069-1 P A B X -B B A -1-069-1 BQL = Below quantitation limit. AQL = Above quantitation limit DF = Dilution Factor 001245 Prnedica-Worcester Project Number PABX-BBA 418-018-.PAGE 1-209 Page 23 APPENDIX A REPRESENTATIVE CHROMATOGRAPHIC DATA - BATCH NO. PABX-1-047-1 0C1246 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-210 Page 24 Peak Area Report Sample Name: 1x Blank Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/2/01 1529 O perator Data H ie Name: 0471001.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX.BBA Instrum ent Nam e: MS7 kietnim ant Vial N um ber 1 Comoound # 1) 2) Compound Name MVL M Vt-04 natenMon Tim a (M in.) 0.00 0.00 iSQ 131 135 * 0 0 DIVA)! M* - The lattar *m* In column Inaleam that manual tmagirto ii -- porfonrod on that compound. Print Date and Time: 1/18/01 1d:A4 Page 1 001247 Primedica-Worcester Project Number PABX-BBA 418-018:PA G E 1-211 Page 25 Data File Acq On Sample Mi sc : D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471001.D Vial: 1 2 Jan 2001 15:29 Operator: : lx Blank Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M {RTE Integrator) T1C:M7t0tJ) 0471001.D PABX BBA.M Thu Jan 18 14:44:41 2001 Page 2 001248 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-212 Page 26 Peak Area Report Sam ple Nam e: 1 X STD 1 N otebook R eferenc e : PABX-BBA-1-047 Date A cq u ire d : 1/2/0115:48 O p e ra to r D ata F ile N am e: 0471002.D Data FHe P ath: D:W SD_DATA\PAB\PABX-BBA\1-047-1\ h a tn im e n t M ethod Nam e: PABXJ9BA Instrum ent Nam e: MS7 Instrum ent V ial N um ber. 2 Com pound* 1) 2) C om pound Name MVL MVL-4 R etention Tim e (M in.) 7.02 7.04 Ion 131 135 Peak Am a M* 155 1282 Peak Arne R atio 120.9E-3 M* The letter *iw* In tN a colurw n k id lra le i m et m anuel kiNgraUon n e e perionw ed e n d ie t com pound Print Date and Time: 1/18/01 14:44 Page 1 001249 Primedica-Worcester Project Number PABX-BBA 418-018:PAGE 1-213 Page 27 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471002.D Vial: 2 2 Jan 2001 15:48 Operator: 1 X STD 1 Inst : MS7 0471002.D PABXBBA.N Thu Jan 18 14:45:12 2001 Page 2 001250 Primcdica-Worcestcr Project Number PABX-BBA 418-018:PAGE 1-212 Page 26 Peak Area Report Sam ple Nam e: 1 X STD 1 N otebook R eference: PABX-BBA-1-047 Data A cquired: 1/2/01 15:48 O perator; Data F ile Nam e: 0471002.0 Data F ile R ath: D:'WSD_DATA\PAB\PABX-BBA\1-047-1\ kia tru m e n l M ethod Nam e: PABXJJBA kistn im e n t Name: MS7 Inatrum ent Vtai N um ber. 2 Com pound* 1) 2) C om pound Name MVL MVL-4 R etention Time IMIn.1 7.02 7.04 JSfl 131 135 p-- fc * " m 155 1282 Peak Area R atio 120-9E-3 M * The lanar *m* In thle cokim n k K celee M iel m anuel n ie gieUon a ae partorm ed on a t com pound. Print Date and Time: 1/18/01 14:44 Page 1 001251 Primedica-Worcestcr Project Number PABX-BBA 418-018:PAGE 1-213 Page 27 Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471002.D Vial: 2 2 Jan 2001 15:48 Operator: 1 X STD 1 Inst : MS7 M V L _______________________________ Amount:_____ 0.00 0471002.D PASXBEAM Thu Jan 18 14:45:12 2001 Page 2 001252 Primedica-Worccster Project Number: PABX-BBA 418-018:PAGE 1-214 Page 28 Peak Area Report Sam ple Nm im : 1 X STD 2 N otebook (M a n n e * : PABX-BBA-1-047 Oate A cquired: 1 /2/0 1 18:08 O perator Data FH* N a irn : 0471003.D Data FH* P ath: D*MSD_DATA\PAB1PABX-BBAM-047-1\ kw tm m e nt M ethod Nam e: PABX_BBA b istiu m a n t Mama: MS7 to stn im e nt V ial N um ber 3 Comoound t) 2) M *Th lattar Com pound Nam* MVL MVL-04 R etention Tim e (M in.) 7.02 7.04 Jsa 131 135 p-- * -- tc 213 1441 Peek A n a Retto 147.8E-3 iwaapartonnatfflwa>cewipBMHtt. Print Dale and Time: 1/18/01 14:45 Page 1 001253 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-215 Page 29 Data Pile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-1\0471003,D Vial: 3 2 Jan 2001 16:06 Operator: 1 X STD 2 last : MS7 HVL._____________ _______________________ Amount : . Ion: 131 (7.020) ;Reap: 213 0.00 0471003.D PABX BBA.M Thu Jan 18 14:45:31 2001 Page 2 001254 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-216 Page 30 Peak Area Report Sam ple Name: 1 X STD 3 N otebook Reference: PABX-BBA-1-047 Date A cquired: 1/2 11628 O perator: Data File Name: 0471004.D Data FNe Path: DAMS0_DATA\PA8\PABX-BBA\1-047-1\ In strum ent M ethod Name: PABX_BBA kw tn im a n t Name: MS7 Instrum ent V ial N um ber 4 Comoound 9 1) 2) Com oound Nam* MVL M V L-04 R eten tio n Tim e fMln.1 7.02 7.04 tea 131 135 Peak Area M* 242 1138 21Z.7E-3 M* The le c e r'm * In th is colum n Indir al a r that m anual Integration e e s psrlo n n se cw e ia t cempeundL Print Date and Time: 1/18/01 14:43 Page 1 001255 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-217 P a g e 31 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471004.D Vial: 4 2 Jan 2001 16:28 Operator: 1 X STD 2 Inst : MS7 MVL__________________________________________________A m o u n t: r Ion: 131 7.023) 0 .0 0 0471004-D PABXBBA.M Thu Jan 18 14:45:51 2001 Page 2 001256 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-218 Page 32 Peak Area Report Sam ple N a irn : 1 X STD 4 Notaboo k R eference: PABX-BBA-1-047 Date A cquired: 1/2101 16:47 O parator. D ata R e am a: 047100S.D Data F la P ath: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ kietrum ant M ethod Nam e: PABX.BBA bm tn in ie nt Name: MS7 toetnim ent V ial N um ber. 5 Com bound 1) 2) C om oound Name MVl MVL-04 R etention Tim a (M in.) 7.02 7.04 IBB 131 135 Peek Am M* 480 1412 Peak Amo R atio 339.9E-3 M* The le n ir *m* In m il cokjm n in d iu m m at m anuel lotegralle n wee pertam iad on B it com oound. Pnnt Dale and Time: 1/18/01 14:46 Page 1 00125? Primedica-Worcester Project Number. PABX-BBA 418-018.PAGE 1-219 Page 33 Data File Acq On Sanple Mise D:\KSD DATA\PAB\PABX-BBA\1-047-1\0471005.D Vial: S 2 Jan 2001 16:47 Operator: 1 X STD 4 Inst : MS7 MVL ___ _____________________________ Amount :_____ 0.00 ! Ion: 131 (7.020) 047100S.D PABXBBA.M Thu Jan IB 14:46:11 2001 Page 2 001258 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-220 Page 34 Peak Area Report Sam ple Name: 1 X STD 5 N otebook Reference: PABX-BBA-1-047 Dele A cquired: 1 /2/01 17:07 O perator. Data FHe Name: 0471006.D Data FMa P ath: DflMSD_DATA\PAB\PABX-BBA\1-047-1\ Inatrum ent Meth od Name: PABX.BBA Inebum ent Name: MS7 Inetnim ent V ial N um ber 6 Com pound 1) 2) Ssm ew nd Nm i MVL M V L44 R etention Tim e (M in.) 7.02 7.04 te a 131 135 Peak Area M* 790 1213 Peak Area Rado 6 5 1 3 E -3 M* a The tetre* ` m* m tM e eekmeiIn d icarne a w l m anual Integrane nee p n f onm d en eue r e iQ M iC Print Date and Tine: 1/18/01 14:46 Page 1 001^59 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE I-22I Page 35 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471006.D Vial: 2 Jan 2001 17:07 Operator: 1 X STD S Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M IRTE Integrator) ........ ......... " - TKf: 0471OOe.D 0471006-D PABXBBA.M Thu Jan 18 14:46:30 2001 Page 2 012C0 Primedica-Worcester Project Number PABX-BBA 418-018:PAGE 1-222 Page 36 Peak Area Report Sam ple Nam e: 1 X STD 6 N otebook R eference: PABX-BBA-1-047 Dele A cq u in o : 1/2/01 17:27 O perator: Data F ile Nam e; 0471007.D Data F ile P ath: D:\MSD_DATA\PAB\PABX-BBA\1-047-U In stru m e n t M ethod Nam e: PABX.BBA Instrum ent Name: MS7 Instrum ent VM Num ber: 7 Comoound i U 2) Comoound Nm m MVL MVL-4 R etention Ion P eakA iea M* Peak A n e R atio 7.02 131 1257 1032E-2 7.04 135 1218 U ' The le tter 'w * In th is cosm er Indic e n e that manuel knegretlcn wee perform ed en brat cornpeundL Print Oste and Tene: 1/1B/01 14:46 Page 1 Q01Z&1 Primedica-Worcester Project Number PABX-BBA 418-018:PAGE 1-223 Page 37 DaCa File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471007.D Vial: 7 2 Jan 2001 17:27 Operator: 1 X STD 6 Inst : MS7 0471007.E PABXBBA.M Thu Jan 18 14:46:50 2001 Page 2 001262 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-224 Page 38 Peak Area Report Sample N aim : 1 X STD 7 Notebook Reference: PABX-8BA-1-047 Data Acquired: 1/2/01 17:47 O p erator Data File N aim : 0471008.D tnatnifm nt N aim : MS7 Data n c Path: D:\MSO_DATA\PAB\PABX-BBA\1-047-1\ tnatnnm nt Vial Number; 8 kiettum ent Method Nemo: PABX.BBA Comoound 1) 2) Comoound Naim MVL MVL-04 Retention Tima (Min.) 7.02 7.04 Jsn 131 135 2166 1284 Peak A im Ratio 168.7E-2 M*t The lattar *m* In m e column millratee eurt manuel integrane m e perfotmed on met rempeund. Print Date and Tane: 1/18/01 14:47 Page 1 001263 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-225 Page 39 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471008.D Vial: B 2 Jan 2001 17:47 Operator: 1 X STD 7 Inst : MS7 Integration Parants : RTEINT.P ant Method : D:\HPCHEM\l\HETHODS\PABX_BBA.M (RTE Integrator) MVL_______________________________________ Amount :______ 0.00 Ion: 131 (7.018) ! R e s p : 2166 1 ..1L : 1-- ''1111' '1' '1! 1' 620 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 720 730 7.40 7.50 7.60 7.70 7.80 7.90 MVL-d4 Amount: 0.00 mi 135 (7.042) A p : 1284 .JV ' 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.S0 7.60 770 7.B0 7.80 0471008.D PABX BBA.M Thu Jan 18 14:47:10 2001 Page 2 001264 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-226 Page 40 Peak Area Report Sam ple Name: 1 X CONTROL Notebook Roteionee: PABX-BBA-1-047 Date Acquired: 1/2/0118:07 O p erato r. Date F ile Nam e: 0471009.D Date FHe Patti: O:\MSO_DATAVPAB\PABX-BBA\1-047-1\ Inatnim enl M ethod N a m : PABX.BBA kntnim em N a m : MS7 Inatnim ant Vial N um ber 9 C om Dound* 0 2) Comoound Name MVL MVL-d4 Retention Tim e IM in-i 0.00 7.04 2 0 131 135 PtaK A rw C . 0 1067 Peak Area Ratio OOO.OE-3 M * T h e tener *m* In m e colum n Indicate te at m anual Integratio n wee perform ed on tria l compound. Print Date and Tene: 1/18/01 14:47 Page 1 001265 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-227 P age 41 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\ 0471009.D Vial: 9 2 Jan 2001 18:07 Operator: 1 X CONTROL Inst : MS7 Integration Parants: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL_______________________________________ Amount : Mot Found Ion: 131 (0.000) 0471009.D PABX_BBA.M Thu Jan 18 14:47:30 2001 Page 2 001266 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-228 Page 42 Peak Area Report Sample Name: 1 X OC LOW Notebook Reterance: PABX-BBA-1-047 D ate A c q u ire d : 1/2/01 10:26 O perator Data File Name: 0471014.D Data File Path: D:\MSD_DATA\PAB\PABX-BSA\1-047-1\ instrum ent Method Name: PABX.BBA biatrum ent Name: MS7 Instrum ent Vial N um ber 14 Comoound # U 2) Com pound Name MVL MVL-04 Retention Tim e (M in.i 7.02 7.04 ten 131 135 Peak Aree M* 184 1135 Peak Area Ratio 162.IE -3 M* D m s t a r "m " In th is colum n in d lealsa that m anual Im agialluei w as partnrm od on that com pound. Print Dale and Tme: 1/16/01 14:46 Page X V 0 12G 7 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 2 Jan 2001 IB:26 Operator: 1 X QC LOW Inst : MS7 0471014.D PABX BBA.M Thu Jan 18 14:49:08 2001 Page 2 001268 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: 1/24)1 18:46 O p erato r. Data FUe Name: 0471025.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ tnitm m ent Method Name: PABX.BBA h a tn iment Name: MS7 Inatnim ent Vial Number: 25 Comoound # 1) 2) Comoound N itn a MVL MVL-d4 Retention Tim e (M in.) 7.02 7.04 Ion 131 135 Peak Area M* 6106 706 Peek Area Ratio 864.9E-2 M* T ire tetter 'm ` In thle colum n In ik ra lca d ie t m enuet lirle g retlon o ae portonwed a n that c om pound. Print Date and Time: 1/18/01 14:53 Page 1 001269 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-231 Page 45 V Data File Acq Dn Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471025-D Vial: 25 2 Jan 2001 18:46 Operator: 10930-GROUP3-FO Inst : MS7 Integration Params: RTEINT-P 0 ^ t Method : D:\HPCHEM\l\MBTHODS\PABX_BBA.M (RTB Integrator) M V L _______ ____ ___________________ _____ A m o unt:______ 0^00 " i o n : '131 (7.015) !Resp: 6106 6.20 6.30 6.40 650 6,60 ' 6.70 6.80 6.90 7.1X1 7.10 7^0 7J0 7.40 7.50 7.60 7.70 7.80 7.90 M'ri,-d4 Amount: 0.00 0471025.D PABX_BBA.M Thu Jan 18 14:53:42 2001 Page 2 0012^0 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-232 Page 46 Peak Area Report Sample Name: 10931-GROUP3-FO Notebook Reference: PABX-B8A-1-047 Dale Acquired: 1/210119.06 O p erator: Data File Name: 0471026.D Data File Patti: D:\MS0_DATA\PAB\PABX-BBA11-047-1\ Instrum ent Method Nam e: PABX.BBA tnatrum ent Name: MS7 Inctnim ent Vial Number: 26 Comoound # it 2) Com oound Nomo MVL MVL-d4 Retention Tim e flUtfn.) 7.01 7.04 ton 131 135 Peak Area M * 5583 567 Peak Area Ratio 984.7E-2 M " * The toner "m ` In into colum n in dicarne m at m anual Intojraeon ama perform ed on m ar r om pound. Phnl Data and Time: 1/18/01 14:53 Page 1 001271 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-233 Page 47 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471026.D Vial: 26 2 Jan 2001 19:06 Operator: 10931-GROOP3-FO Inst : MS7 Integration Params: RTEINT.P it Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) MVL____ ____ ___ _____ _____ r* Ion: 131 (7.014)'" Amount : __0._00 0471026.D PABX SBA.M Thu Jan 18 14:54:02 2001 Page 2 001242 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-234 Page 48 Peak Area Report Sample Nam e: 10932-GROUP3-FO Notebook Retefonce: PABX-6BA-1-047 Date Acquired: 1/2/01 1926 Operator: Data File Nam e: 0471027.0 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Nam e: PABX_BBA Instrum ent Name: MS7 Instrum ent Vial Number: 27 Compound * 1) 2) Comoound Nam * MVL MVL-4 Retention Tim e (Mln.1 7.02 7.04 Ion 131 135 PaakA rea Iff* 5369 792 Peak Area Ratio 680.4E-2 M* The le tte r "m* In O ils cokim n Ind lcatss P is i m anual Integration w as p srtonn sd on tfie t com pound. Print Dale and Time: 1/18/01 14:54 Page 1 001273 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-235 Page 49 yuouLiuoL-AW it \w* iv e v ie v e u Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\ 0471027.D Vial: 27 2 Jan 2001 19:26 Operator: 10932-GR0UP3-F0 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BHA.M (RTE Integrator) 0471027.D PABX BBA.M Thu Jan 18 14:54:22 2001 Page 7 O O I2 7 4 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-1-047 Date Acquired: 1/2/01 19:45 O perator Data Fila Name: 0471028.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ Instnim ent Method Name: PABX_BBA Inatrument Name: MS7 Inatrum ent VW N um ber 28 Comoound # 1) 2) Comoound Name MVL MVU-4 Retention Tim e (Min.) 7.02 7.04 (on 131 135 Peak Aree M* 7053 515 Peek Aree Retto 137DE-1 M* The tener *m ' In th le colum n M lc a te e the! m enuel Inle ip a lim i ne pertonwed en ew t com pound. Pnnl Dale and Time: 1/18/01 14:54 Page 1 001275 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-237 Page 51 KCJAJLL data File Acq On Sample Mise D:\MSD DATA\PAS\PABX-BBA\l-047-l\0471028.D Vial: 28 2 Jan 2001 19:45 Operator: 10933-GROUP!-FO Inst : MS7 Integration Parana: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0471028.D PABX BBA.M Thu Jan IB 14:54:41 2001 Page 2 001276 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-238 Page 52 Peak Area Report S im ple N in w : 10934-GROUP3-FO Notebook Reference: PABX-BBA-1-047 M e Acquired: 1/21 20:05 O perator Data File Name: 0471029.D M a File Path: D:\MSD_DATA\PAB\PABX-6BA\1-047-1\ Instrum ent Method Name: PABX_BBA Instrum ent Name: MS7 Inatnim ant Vial N um ber 29 Comoound 1) 2) Comoound Name MVL MVL*d4 Retention Tim e (Min.1 7.01 7.04 to ll 131 135 Peak Area M* 3213 646 Peak Area Ratio 497.4E-2 M* - The letmr *m* In Pile column Indlceree that manual Integration nee performed en diet compound. Print Dale and Time: 1/18/01 14:54 Page 1 001277 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-239 Page 53 quantitation Report. iqi nevieweoi Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BHA\1-047-1\0471029.D Vial: 29 2 Jan 2001 20:05 Operator: 10934-GROUP3-FO Inst : MS7 Integration Params: RTEIHT.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 001278 Priniedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-240 Page 54 Peak Area Report Sample Nam e: 10935-GROUP3-FO Notebook Rateranca: PABX-BBA-1-047 Data Acquired: 1/2/01 2025 Operator: Data R ie Nam e: 0471030.D Data R ie Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ feutrument Method Nam e: PABX.BBA Inetnim efit Name: MS7 kiatnim ent Vial Number: 30 Comoound 1) 2) Comoound Norm MVt MVL-CJ4 Retention Tim e <Mln.) 7.02 7.04 fea 131 135 E&dsAsi t 6310 596 Peak Area Ratio 105.9E-1 Pnnt Dale and Tin e: 1/18/01 14:55 Page 1 0 0 1 2 -; 9 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-241 Page 55 u u a n c ita tio n K eport uji' tcevieveat Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471030.D Vial: 30 2 Jan 2001 20:2S Operator: 1093S-GROOP3-FO Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTS Integrator) MVL_________________ _____ ______ : Ion: 131 (7.016) Amount :______ 0.00 0471030.D PABX BBA.M Thu Jan IB 14:55:20 2001 Page 2 0012S0 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-242 Page 56 Peak Area Report Sam ple Nam e: 10937-GROUP3-FO N otebook R eference: PABX-BBA-1-047 Date A cquired: 1/2/01 20:45 O perator: Data F ile Nam e: 0471031.D Data File P ath: D:IM SD_DATA\PAB\PABX-BBA\1-047-1\ In stru m e n t M ethod Nam e: PABX_BBA In stn im e n t Nam e: MS7 Inatrum ent V ial N um ber 31 Compound t U 2) Com oound Name MVL MVL-4 R etention Tim e (M in.) 7.02 7.04 jo a 131 135 E sskAas m: 6228 992 Peak Area R atio 627.8E-2 M* a The lerar *m* Mthis column InWrss n Viet manual integration waa pcrtormcd on Mat compound. Print Date and Time. 1/1B/01 14:55 Page 1 001251 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-243 Page 57 yuantitatron Report lui neviexeui Data File Acq On Saitple Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471031 -D Vial: 31 2 Jan 2001 20:45 Operator: 10937-GROUP3-FO Inst : HS7 Integration Params: RTEINT.P m ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVLi Amount: 0.00 0471031.D PABX BBA.M Thu Jan 18 14:55:40 2001 Page 2 0012&2 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-244 Page 58 Peak Area Report Sam ple Nam e: 10940-GROUP3-FO N otebook R aterenca: PABX-BBA-1-047 Data A cq u ire d : 1 /2/01 21:04 O p erator. Data F ile N am e: 0471032.0 Data F ile P a tti: D:\M SD_0ATA\PAB\PABX-BBA\1-047-1\ Instrum ent M ethod N am e: PABX_BBA In stru m e n t Nam e: MS7 tn a tru m e nt V ia l N um ber. 32 Com oound # 1) 2) C om pound Name MVL M V L-d4 R etention Tim e (M in.) 7.02 7.04 131 135 Peak A rsa M* 7133 602 Peak Area R atio 118.5E-1 W~The e te r 'm * ki mlmcolumn indtcsl-- mar manual in fla tio n m n parlcrmad on Siatconyound. Print Date and Tine: 1/18/01 14:55 Page 1 0012S 3 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-245 Page 59 UUailLXLaLXUll ItepUXL lVJ (ICViewtUI Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1 -047-1\0471032.D Vial: 32 2 Jan 2001 21:04 Operator: 10940-GROUP3-FO Inst : MS7 Integration Pararne: RTEINTP ant Method : D:\HPCHEM\l\KETHODS\PABX_BBA.M (RTE Integrator) 0471032.D PABXBBA.M Thu Jan 18 14:56:00 2001 rage 2 0012o4 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-246 Page 60 Peak Area Report Sam ple Nam e: 10930-GROUP3-F1 N otebook R eference: PABX-BBA-1-0X7 D ale A cq u ire d : 1/2/012124 O perator: Data F ile Nam e: 0471033.0 D ata R ie P ath: D:\MSD_DATA1PAB\PABX-BBA\1-047-H In stru m e n t M ethod Nam a: PABX_BBA In a ln im ent Nam e: MS7 In stru m e n t V ia l N um b e r 33 Com Bound # 1) 2) C om oound Name MVL MVL-d4 R etention Tim e (Mln.1 7.02 7.04 Isa 131 135 Fm Ii & m !C 124493 779 Peak Area R atio 159-6EeO M* - The letter'm " In this column Indicates that manual Integration erne pertormed on that compound. P m t Date and Time: 1/18/01 14:56 Page 1 012S5 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-247 Page 61 Uuantitation Report lUT Reviewed) Data File Acq On Sample Mise D:\MSDDATA\PAB\PABX-BBA\l-047-l\0471033.D Vial: 33 2 Jan 2001 21:24 Operator: 10930-GROP3-F1 Inst : MS7 Integration Pararas: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BHA.M IRTE Integratori M**T. - d 4 ________________ ___________ a m o u n t ______ 000 135 (7.040) 779 11I'' 6.20 630 6.40 6.50 6.60 6.90 7.00 7.10 720 7.30 7.40 7.50 7.60 7.70 7.80 7.90 047.033.D PABX BBA.M Thu Jan 18 14:56:20 2001 Page 2 001286 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-248 Page 62 Peak Area Report Sam ple Nam e: 10931-GROUP3-F1 N otebook R eference: PABX-B BA-1-047 Date A cq u ire d : 1/2/01 21:44 O p e ra to r Data F ile Nam e: 0471034.0 Data F ile P ath: D:\MSD_DATA1PAB\PABX-BBA\1-047-1\ Instru m e n t M ethod Nam e: PABX.BBA Instrum ent Nam e: MS7 In stn im e n t V ial N um b e r 34 Com oound # U 2) C om oound Name MVL MVL-4 R etention Tim e (Mln.1 7.02 7.04 ten 131 135 P W k A ff* m 148288 815 Peak Area R atio 181.9+0 M' The letter *m* In m is coturno tnrttr etee that manual Integration wee performed on that compound. Print Date and Time: 1/18/01 14:56 Page 1 0012S7 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-249 Page 63 Quantitation Report iut Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047 -1\0472034 -D Vial: 34 2 Jan 2001 21:44 Operator: 10931-GROUP3-F1 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL M'nJ-d4 Amount: 0.00 Amount : 0.00 0471034. D PABXBBA.M Thu Jan IB 14:56:39 2001 Tage 2 0012S 8 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-250 Page 64 Peak Area Report Sam p)* Nam: 10932-GROUP3-F1 N otebook Reference: PABX-BBA-1-047 D ato A c q u in o : 1/2/01 22:03 O p erato r. Data FU* Nama: 0471035.D Data Fite P a tti: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ kia tru m e n t M ethod Name: PABX.BBA kw tn im a n t Nama: MS7 kiatrum e n t V ial N um ber: 35 Comoound * 1) 2) C om oound Name MVL MVL-4 R etention Tim e /M in.) 7.02 7.04 ten 131 135 Peak Area M* 44011 655 Peak A n a R atio 671.9E-1 te" The tettar *m" In this column indiala* that manual Integration *e* parlsnnad on teal compound. Print Date and Time: 1/18/01 14:56 Page 1 00129 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-251 Page 65 uuantitation Report mot itevieweai Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047 -1\0471035 .D Vial: 35 2 Jan 2001 22:03 Operator: 10932-GROUP3-F1 Inst : MS7 k3 Integration Pararne: RTEINT.P Puant Method : D:\HPCHEM\1\METH0DS\PABX BBA.M (RTE Integrator) TIC:0471035.0 MVL Amount: 0.00 0471035.0 PABX BBA.M Thu Jan 18 34:56:59 2001 Page 7. 001290 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-252 Page 66 Peak Area Report Sam ple Nam e: 1 X OC MIO N otebook R eference: PABX-BBA-1-047 Data A cquired: 1/2/01 2223 Opera to r: Data F ile Nam e: 0471012.D Data F ila P a th : D:\M SD_0ATA\PAB\PABX-BBA\1-047-1\ In stru m e n t M ethod Nam e: PABX_BBA Instrum ent Name: MS7 In stru m e n t V ia l N um ber 12 ComDound # 1) 2) Comoound Nam* MVL M VL44 R etention Tim a iM In .l 7.02 7.04 JOQ 131 135 Peak Area M* 522 1016 Peak Area R atio 512.8E-3 U* The Uttar *m* m thla column Indicata that manual Integration ama performed on that compound. Prml Dale and Time: 1/18/01 14:48 Pcge 1 001251 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-253 Page 67 Wuauvitauiuu Mviewcut Data File Acq On Sample Mise D:\MSD DATA\PAB\PAB2C-BBA\l-047-l\0471012.D Vial: 12 2 Jan 2001 22:21 Operator: 1 X QC MID Inst : MS7 Integration Params: RTEINT.P it Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTS Integrator) 0471012.D PAfiX bflA.K Thu Jan 18 14:48:29 2001 Page 2 001292 Primedica-Worcester Project Number: PABX-BBA 418-018 :PAGE 1-254 Page 68 Peak Area Report Sam ple Name: 10933-GROUP3-F1 N otebook Reference: PABX-BBA-1-047 Date A cq u ire d : 1/2/01 22:43 O p e ra to r Data File Name: 0471036.D Date H ie Path: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ In stru m e n t M ethod Name: PABX_B8A kia tn im e n t Nam e: MS7 kia tn im e n t V ial N um b e r 36 Comoound i) 2) C om oound Name MVL M VL-04 R etention Tim e (M in i 7.02 7.04 on 131 135 Sss&Aos m 106656 m 763 Peak Area R atio 136.2E+0 M* > The le u r "m` m this column Indicete that manual Imagielkm u s performed en that compound. Pimi Date and Time: 1/18/01 14:57 Page \ 001293 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-255 Page 69 Uuancitacion Keporc lUT 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-GROUP3 -FI Inst : MS7 5 Integration Params: RTEINT.P lant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) 710:0471036.0 MVL__________________________________Amount:______0.00 Ion: 131 (7.016) :Resp: 106656 MVL-d4 Amount: 0.00 0471036.D PABX BEA.M Thu Jan 18 14-57:19 2001 Page 2 001294 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-256 Page 70 Peak Area Report Sam ple Nam e: 10934-GROUP3-F1 N otebook R eference: PABX-8BA-1-047 Date A cq u ire d : 1 /2/01 23:02 O p e ra to r D a ta n te Nam e: 0471037.D Data F ile P a tti: D:\MSD_DATA\PAB\PABX-BBA\1-047-1V In itru m e n t M ethod Nam e: PABX.BBA tn atm m ent Nam e: MS7 tn a tru m e nt V ia l N um b e r 37 ComDOund # 1) 2) ' C om pound Nama MVL M VL-d4 R etantion Tim e fM ln .l 7.01 7.04 2 0 131 135 Peak Area y 46551 659 Peak Area R atio 706.4E-1 M** The leasr'in* In this column Intficataeeiat manual hitsgrattan m s partonnad on that compoundL Print Dale and Time: 1/18/01 14:57 Page 1 001295 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-257 Page 71 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471037 .D Vial: 37 2 Jan 2001 23 :02 Operator: 10934-GROUP3-F1 Inst : MS7 Integration Params: RTEIHT.P kant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) TIC: 0471037.D t C471037.D PABXBBA.M Thu Jan 18 14:57.^8 2001 Page 2 001296 Primedica-Worcester Project Number: PABX-BBA 418-018.-PAGE 1-258 Page 72 Peak Area Report Sam ple Nam e: 10935-GROUP3-F1 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/2/01 23:22 O perator: Data F ile Nam e: 0471038.D Data F ile P a th : D:\MS0_DATA\PAB\PABX-BBA\1 -047-1\ Ina tru m e nt M ethod N am e: PABX_BBA kistm m a nt Nam e: MS7 In a tn im e n t V ial N um ber 38 Compound 1) 2) C om oound Name MVt M VL-d4 R etention Tim e fM ln .l 7.02 7.04 Ion 131 135 Peak Area 1C 85000 B&4 Peak Area R atio 950.8E-1 II* The letter *m * In thla column Indicate* that manual Intgration area performed an that compound. Prinl Date and Time: 1/18/01 i 4:57 Page 1 G GrUrJ V Primedica-Worcester Project Number: PABX-BBA 418-018:PA GE 1-259 Page 73 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047 -1\04 71038 .D Vial: 38 2 Jan 2001 23:22 Operator: 1 0 9 3 5 -GROUF3-Fl Inst : MS7 0471038.D PABX B5A.M Thu Jan ie 14:57:-i0 2001 Paqtr 001238 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-260 Page 74 Peak Area Report Sam ple Nam e: 10937GROUP3-F1 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1 /2/01 23:42 O perator. Data F ile Nam e: 0471039.D Data F ila P ath: D:\MSD_DATA\PAB\PABX-BBAM -047-11 In stru m e n t M ethod Nam e: PABX_B8A b iatrum ent Nam e: MS7 In stru m e n t V ia l N u m b e r 39 Comoound 1) 2) C om oound Name MVL M VL-04 R etention Tim e (M in.) 7.02 7.04 fern 131 135 Peak Area M* 76789 751 Peak Area R atio 102.2E+0 M* The la ra r *m * In this column kirtlu taa that manual knay itlo n nee performed on that compound. : -kit Dale and Time: 1/18/01 1:58 Page 1 Primedica-Worcester Project Number: PABX-BBA 418-018.PA G E 1-261 P age 75 Data File Acq On Sample Hisc D:\MSDDATA\PAB\PABX-BaA\l-047-l\0471039.D Vial: 39 2 Jan 2001 23:42 Operator: 10937GROP3-F1 Inst : MS7 KVL : Ion: 131 '(7.017) .Amount:. 0 .JLQ 0471039.D PABX BBA.M Thu Jan 18 14:58:18 2001 Pace 2 O O IO O O Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-262 Page 76 Peak Area Report Sam ple Nam e: 10940-GROUP3-F1 N otebook R eference: PABX-6BA-1-047 Date Acquired: 1/3/01 0:01 O p erator: Data Fite Nam e: 0471040.D Data Fite P a th : D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ In stru m e n t M ethod Nam e: PABX_BBA b iatm m ent Nam e: MS7 In ttn im e n t V ial N um ber. 40 Comoound # 1) 2) C om oound Name MVL M VL-d4 R etention Tim e (M ln.l 7.02 7.04 Ion 131 135 Peak Area IT 73060 712 Peak Area R atio 102.6E+0 M * The tetter "m" In Pile column Indicates Piet manuel InregraMon ewe pstterm ed on that compound. Preti Dale and Time: 1/18/01 14:58 Page l 001301 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGEI-263 Page 77 Data File Acq On Sample Mise D:\MSD DATANPAB\PABX-BBA\1 -047-1\0471040.D Vial: 40 3 Jan 2001 00:01 Operator: 10940-GROUP3-F1 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL___________________________ _________ Amount : Ion: 131 (7.016) ... " 0.00 0471040.D PABX BBA.M Thu Jan 19 14:58:38 2001 Page 2 001302 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-264 Page 78 Peak Area Report Samp* N am *: 10929-GROUP3-F0 N otebook R *i* r*n c * : PABX-BBA-1-047 Data A cq u ire d : 1/3/01 021 O perator. Data R ie Nam e: 0471041JD Data R ie P e tti: 0:W ISD_DATA\PAB\PABX-BBA\1-047-1\ In stru m e n t M ethod N am *: PABX.BBA feietnim ent Nam e: MS7 Inetrum ent V ia l N um b e r 41 Comoound 1) Z) C om oound Name M Vl MVL-CJ4 R etention Tim e IM In .l 7.02 7.04 Ion 131 135 Peak Area M* 1190 1108 Peak Area R atto 107.4E-2 M * The Iettar *in * In th la c o k m i Indicata ! that manual intagialien ess partermed en that compound. Prim Date and Tme: 1/1B/01 1 <*:r n Pa,e 1 001303 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-265 Page 79 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047.1X0471041.D Vial: 41 3 Jan 2001 00:21 Operator: 10929-GROUP3-F0 Inst : MS7 ? Integration Parana: RTEINT.P L a n t Method : D:\HPCKEM\1\METH0DS\PABX_BBA.M (RTE Integrator) M71041.D PABX_BBA M Thu Jan 18 1^:58:58 2f-0J. Page 2 001304 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-266 Page 80 Peak Area Report Sam ple Name: 1093B-GROUP3-F0 N otebook R eference: PABX-8BA-1-047 Date A cq u ire d : 1/3/01 0:41 O perator. Data F ile Name: 0471042.D Data R ie P ath: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ kia tru m e n t M ethod Name: PABX.BBA In stru m e n t Nam e: MS7 kiatrum e n t V ial N um b e r 42 Comoound # 1) 2) C om oound Name MVL M V L-d4 R etention Tim e /Min.1 7.02 7.04 SB 131 135 Peak A n a M* 1604 917 Peak Area R atio 174SE-2 M - a The lener *m * In thle column indicates that manual Integration ama performed on that confound. Print Date and Time: 1/18/01 14:59 Page 1 001305 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-267 Page 81 tW* i v e i i o c u i Data Pile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\04 71042 .D Vial: 42 3 Jan 2001 00:41 Operator: 10938-GROUP3-F0 Inst : MS7 Integration Pararne : R T E I N T .P t Method : D:\HPCHEM\l\METHODS\PABX_BEA.M (RTS Integrator) 0471042. D PABX E B A .M Thu Jan .`.8 14:59:?'/ 2001 Pb^fc 2 001306 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-268 Page 82 Peak Area Report Sam ple N am *: 10942-QROUP3-FC N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 1:00 O perator. Data F ile Nam e: 0471043.0 Data FU* P ath: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ ta stn im e nt M ethod Nam e: PABX_BBA In stn im e n t Nam e: MS7 In stru m e n t V ia l N um ber: 43 Comoound # 1) 2) Com oound Nam* MVL MVI-CU R otentton T im * (Mln.1 7.02 7.04 io n 131 135 Peek Aree M* 1938 777 Peak A ra i R atio 249.4E-2 U * The n itc fm * In il column Inditelo diet manuel InMgremiii wee poeforinod on Mia compound. Print Date and Time: 1/B/OI 14:59 Page 1 001307 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-269 Page 83 Data File Acq On Sample Mise quantitation Report aeviewea; D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471043 .D Vial: 43 3 Jan 2001 1:00 Operator: 10942-GROUP3-F0 Inst : MS7 MVL Amount: 0.00 0471043. D '.UiX BBA.M Thv Jan 16 14-59:37 2001 Page 2 001308 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-270 Page 84 Peak Area Report Sam ple Nam e: 10941-GROUP3-F0 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 150 O perator: Data File Nam e: 0471044.D Data F ile P a tti: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ In strum ent M ethod Nam e: PABXJ3BA Inatm m ent Nam e: MS7 In a tru m e nt V ia l N um ber: 44 Comoound f U 2) C om pound Name MVL MVL-4 R etention Tim e (M in.) 7.01 7.04 Ion 131 135 Peak Area M* 4575 1658 Peak Area R atio 275.9E-2 M * The letter *m * In B i l l column Indicates that manual Integration ana perf ormed on that compound. Print Date and Time: 1/18/01 14:55 Page 1 0013C9 Primedica-Worcester Project Number: PABX-BBA 418-018:PA G E 1*271 Page 85 V u a itL i L a L iu u a c ^ u i c iy i R ev A cw eu / Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471044 .D Vial: 44 3 Jan 2001 1:20 Operator: 10941-GROUP3-F0 Inst : MS7 Integration Pararne: RTEINT.F ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTS Integrator) kcurawte ' "' ' TIC:0471044.0 MVL Amount : 0.00 0471044.D 'ABX BBA.M Thu Jan 18 .'4:59:57 200 Page 2 001310 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-272 Page 86 Peak Area Report Sam ple Nam e: 10929GROUP3-F1 N otebook R eference: PABX-BBA-1-047 Data A cquired: 1/3X11 1:40 O perator. Data R ia Nam e: 0471045.D Data R ia P a th : O:\MSD_DATA\PAB\PABX-BBAW -047-1\ In stru m e n t M ethod Nam e: PABX_BBA In o tn im en t Name: MS7 Ina tm m a n t V ia l N um ber 45 Comoound 4 1) 2) Comoound Nam* MVL MVL-4 R etention Tim e (M in.) 7.02 7.04 te a 131 135 P u k Ama M* 1254 1183 Peek Area R atio 106.0E-2 M * > The letaar *m * In Vila column Indicarne that manual IriiayeU tm nua partnrmad an VuH compound. Prati Dali and Time: 1/18/01 15:00 Page 1. 001311 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-273 Page 87 y u O u b A b O w A w u (\e p w ii b Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471045.D Vial: 45 3 Jan 2001 1:40 Operator: 1092 9GROUP3-Fl Inst : MS7 Integration Params: RTEINT.P t Method : D:\HPCHEM\l\METHODS\PABX BBA.M (RTE Integrator) MVL_________ ____________ __ _____ ___ __ Amount :______ 0.00 Ion: *131 (7.06) ' I ' ' I ,- r I i 1,1 I r ! - T ., ( . p r-f j I 6.20 6.30 6.40 6.50 6.60 6.70 6-BO 6,90 7.00 7.10 7.20 730 7.40 7.50 7.60 7.70 7.80 7.90 0471045.D P:\BX BBA.r Thu Jan 18 15:00:16 2001 Page 2 001312 Primedica-Worcester Project Number: PABX-BBA 418-018 .'PAGE 1-274 Page 88 Peak Area Report Sam ple Nam e: 10942-GROUP3-P1 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 1:59 O p e ra to r D ata R le Nam e: 0471046.D Data FNe P ath: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ kia tn im e n t M ethod Nam e: PABX.BBA In ctiu m e n t Name: MS7 in stru m e n t V ia l N um b e r 46 Comoound U 2) C om oound Nomo MVL MVL-4 R etention Tim e fM inJ 7.02 7.04 ton 131 135 Peak A rea M ' 2074 882 Peak A rea R atio 235.1E-2 I t " Ttie tener Vn" In thiecolunin indicetee that menuel Integreden aac performed on bie t compound. Print Date and rime: 1/1B/01 15:00 *nge 1 001313 Priniedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-275 Page 89 Daca File Acq On Sample Mise iv c p v ii l vw* * **** D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471046.D Vial: 46 3 Jan 2001 1:59 Operator: 10942-GROUP3 *FI Inst : MS7 0473.046.D PABX BBA .M Thu Jan 18 15:00:36 200 ; q e 2 001314 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-276 Page 90 Peak Area Report Sam ple Nam e: 1 X OC HIGH N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 2:19 O p e ra to r Data F ile N am e: 0471010.0 Data R ie P ath: D:\M SD_0ATA\PAB\PABX-BBA\1-047-1\ Instrum ent M ethod Nam e: P ABX.B BA tnatnw nent Nam e: MS7 Inetm m ent V ia l N um b e r 1 0 Compound # 0 2) C om oound Nmtm MVL M V L-d4 R etention Tim e /M in.) 7.02 7.04 ton 131 135 Peak Am a M* 1470 1091 Peak Area R atio 134.7E-2 M ' The te tta r-m " In this column fmftcatss mat manual integration was performed on that compound. Pnnt Date and Time: 1/18/01 14:47 Page ; 001315 Primedica-Wofcester Project Number: PABX-BBA 418-018:PAGE 1-277 P a g e 91 UUaubALauAUii iv* ftcticucui Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471010.D Vial: 10 3 Jan 2001 2:19 Operator: 1 X QC HIGH Inst : MS7 Integration Parame: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) 0471010.D i'ABX BBA.M Thu Jan 10 14:47:49 2001 Page 2 001316 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-278 Page 92 Peak Area Report S im p le Nam e: 1O945-GROUP4-F0 N otebook R eference: PABX-BBA-1-047 Date Acquired: 1/3/01 2:39 O p erator: Data File Nam e: 0471047.D Data F ile P ath: D:\MSD_DATA\PABVPABX-BBA\1-047-1\ In stru m e n t M ethod Name: PABX.BBA In stru m e n t Nam e: MS7 In stru m e n t V ial N u m b e r 47 Compound # i) 2) Com pound Nam* MVL MVL<M R otontion Tim e (M in.) 7.02 7.04 JOQ 131 135 Peak Area M* 6392 SOS Peak Area R atio 126.6E-1 M ' The tener -m * in this column Indicates met manual Integration teas pariormad on mat compouraL Print Date and Time: 1/18/01 15 00 'age 1 001317 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-279 Page 93 l/uantication nepuiL ivi nvweui Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471047 .D Vial: 47 3 Jan 2001 2:39 Operator: 1094S-GROUP4-F0 Inst : MS7 Integration Parame: RTEINT.P liant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0471047 r; pABX Thu Jan j1 15:00:'''. 2001 Page. J 001318 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-280 Page 94 Peak Area Report Sam ple Nam e: 10946-GROUP4-F0 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 2:58 O p e ra to r Data FHe Nam e: 047048.0 Data F ile P ath: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ m atrum ent M ethod Nam e: PABX-BBA kw tn un e n t Nam e: MS7 In stn im e nt V ial N um ber. 48 C om oourtd # K 2) C om ooundN am e MVL MVL-4 R etention T im e fMln.1 7.02 7.04 ten 131 135 Peak A n a M* 9124 909 Peak Area R atio 100.4E-1 M* - The lener "m* In eile cnlunei Indicare Orel manual ImepreMan eiee pertormea en a t eomecrard. P m l Date and Timr: 1/18/01 K : 4 Paar 1 001319 Primedica-Worcester Project Number: PABX-BBA 418-018:PA GE 1-281 Page 95 guancxcacion xeport vyi neviisweui Daca File Acq On Sample Mise D:\MSD DATANPABNPABX-BBAX1-047 -1N04704 B.D Vial: 48 3 Jan 2001 2:58 Operator: 10946-GROUP4-F0 Inst : MS7 Integration Parama: RTEIHT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 047046 O PAEX HBA.M Thu Jan IB 14:44:73 2001 Pat_f 2 00130 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-282 Page 96 Peak Area Report Sam ple N a m : 10947-GROUP4-F0 N otebook R eterence: PABX-BBA-1-047 Data A cquired: 1/3/01 3:18 O p e ra to r Data F ile Name: 0471049.D Data F ile P ath: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ kiatrum e n t M ethod N a m : PABX.BBA kiatrum e n t N a m : MS7 kia d u m e n l V ial N um b e r 49 Com oound* 1) Z) C om oound Nomo MVL MVL-4 R otontion Tim o (M in.) 7.02 7.04 2 2 131 135 Peak Am a M* 9253 968 Peak A m R atio 936.5E-2 M' m The le tto In th is column indic te e th a t manual Intepraden a m pertormed en diet compound. Ptinl Dale and Time: 1/18/01 15:01 Paje 1 001321 Primedica-Worcestcr Project Number: PABX-BBA 418-018:PAGE 1-283 Page 97 Vuuiuiwawxwi ivevieiieuf Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1 -047-1\0471049.D Vial: 49 3 Jan 2001 3:18 Operator: 10947-6ROUP4-F0 last : MS7 Integration Parame: RTEINT.P ant Method : D:\KPCHS4\l\METKODS\PABX_BBA.M (RTE Integrator) Mvu-d4 Amount: 0.00 0471041 > PABX BBA.M Thu Jan IB 15:01:16 2001 Page 3 001322 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-284 Page 98 Peak Area Report S am ple N am e: 10949-GROUP4-F0 N otebook R eferen ce: PABX-BBA-1-047 D ate A cq u ired : 1/3/01 3:37 O perator D ata File N am e: 0471050.D D ata File P a th : D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ inetnim ent M ethod N am e: PABX.BBA Inetnim ent Nam e: MS7 k w tru m e n t Vial N um ber. 50 C om oound* 1) 2) C om oound Name MVL MVL-4 R eten tio n Tim (Min.) 7.01 7.04 ba 131 135 P e a k A rea M* 4356 047 P eak Area Ratio 4 6 0 2 E -2 Pnnt Date and Tene: 1/18/01 15:01 Page 1 001323 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-285 Page 99 Uuantitation Report: tur Keviewea/ Data File Acq On Sample Mise D:\MSD nATA\PAB\PABX-BBA\l-047-l\0471050.D Vial: 50 3 Jan 2001 3:37 Operator: 10949-GROUP4-FO Inst : MS7 Integration Parants: RTEIMT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M IRTE Integrator) 0471G50.D PABX BBA.M Thu Jan 18 15:0'.36 2001 Page 2 001324 Primedica-Worccstcr Project Number: PABX-BBA 418-018:PAGE 1-286 Page 100 Peak Area Report Sample Ham : 10950-GROUP4-F0 Notebook Retefanca: PABX-BBA-1-047 Data Acquired: 1/3RJ1 3:57 O p erato r. Data FUa Nama: 0471051.0 Data Fila Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ tnatnim ent Method Name: PABXTJBA tnatnim ent Kama: MS7 kwtm m ent Vial N um ber 51 Comoound f 1) 2) Comoound Name MVL MVL-4 Retention Tim e (M inJ 7.02 7.04 Is a 131 135 Peek Aim M* 17179 1993 Peak Arne Ratio 862.0E-2 1 18 1Prim Date and Time: / /ri 15:01 Page 1 001325 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-287 Page 101 uuantication Keport \\fi nevieweay Data Pile Acq On Sample Mise D:\MSD QATA\PAB\PABX-BBA\l-047 -1 \0471051.D Vial: SI 3 Jan 2001 3:57 Operator: 10950-GROUP4-F0 Inst : MS7 Integration Params: RTEIN7.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0471051.D PABX BBA.M Thu Jan 8 15:03. 'i'> 2001 Pari: 2 001326 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-288 Page 102 Peak Area Report Sam p it Nam e: 10951-GROUP4-F0 N otebook R ateranca: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 4:17 O p e ra to r Data Fite Nam e: 0471052.D Date Fite P ath: D:\M S0_DATA\PAB\PABX-BBA\1-047-1\ kia tn im e n t M ethod Nam e: PABX_BBA Inatrum en N am e: MS7 Instrum ent V ia l N u m b e r 52 Com oound# 1) 2) C om oound Name MVU MVL-4 R etention T im a /M in.) 7.02 7.04 lea 131 135 P te kA re e Ml 10058 2193 Peak A rea R atio 823.48-2 M* Tha Utter `m* in ite column Indicates th at manual InregraiJaii was pertormed an that compound. Print Date and lim e : 1/18/01 1S:02 a g e 1 001327 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-289 Page 103 Data File Acq On Sample Mise yuantxtation Report iyx Kevxeweaj D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471052.D Vial: 52 3 Jan 2001 4:17 Operator: 10951-GROUP4-F0 Inst : MS7 > /1052 .D PABX BBA.M Thu Jan 18 '<5:02:16 ''.U01 Page 2 001328 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-290 Page 104 Peak Area Report Sam p Name: 1095343 ROUP4-F0 Notebook Reference; PABX-BBA-1-047 Date Acquired: 1 3 0 1 4:36 O perator Date FHe Name: 0471053.D Data FHe Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ Instrum ent M ethod Name: PABX.BBA Inatnim ent Name: MS7 Inatnim ent Vial Num ber. 53 Comoound # i) 2) Comoound Name MVl MVL-d4 R etention Tim e (Min.1 7.02 7.04 isa 131 135 PeakA rea M* 12921 1584 Peak Area Ralle B15.7E-2 M* &The letter "m" m this column Indicetee thet manual eneflradon wee performed on m et eompound. P :n t Date and Tb.ie: 1/18/01 15:02 P. . 1 0C13A9 Primedica-Worcester Project Number: PABX-BBA 418-018.-PAGE 1-291 Page 105 VuautiLALiuu Reputi tut n e v i B w e u / Data File Acq On Sample Mise D:\HSD DATA\PAB\PABX-BBA\l-047-l\0471053.D Vial: S3 3 Jan 2001 4:36 Operator: 109S3-GROUP4-F0 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M IRTE Integrator) 047105 0 PABX_BSA.M Thu Jan JB 15:02:35 2001 Pag< 001330 Primedica-Worceiiier Project Number: PABX-BBA 418-018:PAGE 1-292 Page 106 Peak Area Report Sam ple Name: 10954-GROUP4-f0 Notebook Retefence:PABX-BBA-1-047 Date Acquired: 1/3/01 4:56 O p erator: Dato R ie Name: 0471004.0 Data FUe Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent M ethod Name: PABX.BBA m atnirnent Name: MS7 Instrum ent V W Num ber. S4 Comoound # O 2) Compound Name MVL MVL*d4 Retention Tim e (M in.) 74)2 7.04 lOQ 131 135 Peak Area M* 11975 1880 Peak Area Ratio 637.0E-2 M*- The Iettar "m` In Otis column Indicates Sat manuel kaeqroMon wee pertermeo on that compound. P m t Date and 1 ime: 1/18/01 10:02 'age 1 001331 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-293 Page 107 Vuautlkabiwu ncpulk w * ivcyiewcui Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471054.D Vial: 54 3 Jan 2001 4:56 Operator: 10954-GROUP4-F0 Inst : MS7 k; Integration Pararas: RTEINT.P Lant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0`. '.054.D PABX BBA.M Thu Jan 18 15:02:55 2001 'age 2 001332 Primedica-Worcester Project Number: PABX-BBA 418-018.PAGE 1-294 Page 108 Peak Area Report Sample Nam e: 10945-GROUP4-F1 Notebook Reference: PABX-BBA-1-047 Data Acquired: 1/301 5:15 O p erator: Data R ie Nam e: 0471055.D Data File Path: D:1MSD_DATA\PAB\PABX-BBA\1-047-1\ kietnim ent Method Nam e: PABX.BBA kietnim ent Name: MS7 kiatnim ent Vial Number: 55 Comoound 21)) Comoound Name MVL MVL-4 Retention Time iMin.l 7.01 7.04 Sea 131 135 Peak Area M* 2B5856 1720 Peak Area Ratio 1662E+0 M*Tho totter 'in" in ihto column Indksateo that m partermedon that corraround. P rin t Date and T in s: i/18/01 15:03 Page 1 001333 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-295 Page 109 Data File Acq On Sample Mise UUttUCJ.U4l.aUXl Kttpuau VWJ ttevxeweuu D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471055.I> Vial: 5S 3 Jan 2001 5:15 Operator: 10945-GROUP4-F1 Inst : MS7 MVL _ _ ___ _______________________ Aiaount :______ 0.00 , Ion: *131 "(7.03) ' Reap: 285856 0471055.D PABXBBA.M Thu Jan 18 15:03:15 1001 Page : 001334 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-296 Page 110 Peak Area Report Sample Name: 10946-GROUP4-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 5:35 Operator: Data R ie Name: 0471056.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Inatrum ent Method Name: PABX_BBA tnatnim ent Name: MS7 tnatnim ent Vial Num ber. 56 Comoound # U 2) Comoound Name MVL MVL-04 R ete n tio n Tim e fM ln.) 7.02 7.04 Ion 131 135 Peak Area M* 380925 2454 Peak Area Ratio 155.2-r0 M*>Thelattar'm*InPilacolumnIndir-alaathatmanualtnlayatlon a m parrermadonWatcompound. Print Ppte and Time: /18/01 15:03 Page 1 001335 Primedica-Worcester Project Number: PABX-BBA 418-018:PA GE 1-297 Page 111 U uauk A u ai.ju ij jiepoi.b. w* K * ic su j Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471056.D Vial: 56 3 Jan 2001 5:35 Operator: 10946-GROUP4-F1 Inst : MS7 Integration Params : RTEINT.P t Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) pn^j-da Amount: 0.00 04710S6.D PABX BBA.M Thu Jan 18 15:'\'*:35 200 .-ge 2 001336 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-298 Page 112 Peak Area Report Sam ple Nam e: 10947-GROUP4-F1 N oteb o o k R o te n n e o : PABX-BBA-1-047 D ate A eq u irad : 173/01 5:55 O p*rato n D ate Fite N am e: 0471057.D D ate Fite P a th : D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent M ethod N ame: PABX_8BA h a tn im e n t Nam e: MS7 k w tn im e n t Vial N u m b er. 57 Compound * 1) 2) MVlC o m p o u n d N am e MVL-4 R eten tio n Tim# (MlnJ 7.01 7.04 ton 131 135 PatLAas 1C 180712 1824 Peek Arne Redo 990.7E-1 M* The tettar *m" In thta eofcimn Indicata* that manual tntay ebon d m performed on that compound. Print Date and Tene; 1/18AH 15:03 lge 1 001337 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-299 Page 113 Data File Acq On Sample Mise yuantltacion xeporc ivi Reviewedi D:\MSD DATA\PAB\PABX-BHA\1-047-1\0471057.D Vial: 57 3 Jan 2001 5:55 Operator: 1 0947-GR0UP4-FI Inst : MS7 0471057.D PABX BBA.M Thu Jan 18 15:'13:55 2001 )age 2 001338 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-300 Page 114 Peak Area Report S am ple Kama: 10949-GROUP4-F1 N otebook R ataran ca: PABX-BBA-1-047 Data A cquired: O perator 6:14 Data R ie Name: 0471058.0 D ata R ie P ath : D:\MSD_DATA\PAB\PABX-BBAU-047-1\ Instnim ofit M ethod Name: PABX.BBA Instrum ent Name: MS7 Instnim ant Vial N um ber. 58 Comoound * 1) 2) C om oound Name MVL MVL-d4 R eten tio n Tim e (Mln.t 7.02 7.04 js a 131 135 E rkA m m: 230169 1560 P eak A rea Ratio 147.5E+0 M* > The lattar *m" In this column Indcalas that manual integration aaa partormed en that confound. P m t Oats and Time: 1/18/01 15:04 Page 001339 Primedica-Worcester Project Number PABX-BBA 418-018-.PAGE 1-301 Page 115 Ouantication Report iut Hevieweai Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-07-l\0471058.D Vial: 58 3 Jan 2001 6:14 Operator: 10949-GROP4-F1 Inst : MS7 a3 Integration Parama: RTEINT.P liant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL________________ __________ _________ _____________0. 00 Ion: 131 (7.017) : Reap: 230169 0471058.D PABXBBAM Thu Jan 18 *:04:IS 2CMJ Page ; 001340 Primedica-Worcester Project Number: PABX-BRA 418-018:PAGE 1-302 Page 116 Peak Area Report S am p le Nam *: 2 X OC LOW N o teb o o k R efe re n ce: PABX-BBA-1-047 D ata A cq u ired : 1/3/01 6:3 Operator: D ata Fit* Nam e: 047101S.D D ata FUa P a th : O:\MSD_DATA\PAB\PABX-BBAW-047-1\ In stru m en t M ethod N am e: PABX_BBA Inatrum ent N am e: MS7 In stru m en t Vial N um ber. 15 Comoound 1) 2) Com oound Name MVL MVL-d4 R eten tio n Tim * /Min.1 7.02 7.04 )on 131 135 P e a k A rea M* 502 1343 Peak Arsa Ratio 373.8E-3 M* The Uttar *m* In ttrt* column Indcalas Otamanual Sway aMon was partncmad en Biat tompound. 8/01Prim Dale and Time: /! 14:49 Pape /. 001341 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-303 Page 117 Quantitation Report IQT Revieweai Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\ 1-047-1\0471015.D Vial: IS 3 Jan 2001 6:34 Operator: 2 X QC LOW Inst : MS7 Integration Pararas: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0471015.D PABX BBA rt Thu Oan 18 1- :49:28 ? . U 1 Page 2 001342 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-304 Page 118 Peak Area Report Sample Name: 10950-GROUP4-F1 Notebook Retatene*: PABX-BBA-1-047 D ett Acquired: 1/3/01 6:54 Operator: Onta Re Neme: 0471059.0 InetniiMnt Name: MS7 Dele FU* Patti: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrument Vial Num ber 59 Instrument Method Name: PABX..BBA Comoound 1) 2) Compound Name MVL MVL-d4 Retention TkneiMln.i 7.01 7.04 lOQ 131 135 Peak Are* Ml 296676 2623 Peak Area Ribo 113.9E+0 M*mThe lerar *m* In eu* column In tftemss taer manuel Imserellen wee performed on Bist eemecund. Pnnl Date raid Time: 1/16/01 15:04 Page : 001343 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-305 Page 119 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1X0471059-D Vial: S9 3 Jan 2001 6:54 Operator: 10950-GROUP4-F1 Inst : MS7 Integration Params: RTBINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) K V L _ _____ ________________ _____ Amount :________ 0.00 Io n i'T i l ` (7 .0 1 5 ) ; Resp: 298676 047105T.O PABX BBA.M Thu Jar '8 15:04:15 2001 001344 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-306 Page 120 Peak Area Report Sample N airn : 10951-GROUP4-F1 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 7:13 O perator Date File Nam e: 0471060.0 Date Fite Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX_BBA InstrumentName: MS7 biatnim ent Vial N um ber 60 Comoound * 1) 2) Comoound Name MVL MVL-d4 Retention Time (Min.1 7.02 7.04 ion 131 135 Peak Area M* 292458 2139 Peak A n a Ratio 136.76+0 M' The tetter *m* Hi this cokimn Indicates that manual Integration was performed on that compound. Prr. Date and Time: 1/18/01 i: :04 Page 1 001C45 Primedica-Worcester Project Number. PABX-BBA 418-018-.PAGE 1-307 Page 121 Quantitation Report IUT Reviewed) Data File A cq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471060 .D Vial: 60 3 Jan 2001 7:13 Operator: 10951-GROOP4-F1 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHBM\1\METHQDS\PABX_BBA.M (RTE Integrator) MVL__________ __________________________ Amount:______ 0-00 i" Ion: 131 (7017) 0 4 'i 1 0 6 0 .D ->ABX BBA.M Thu Jan 18 04:S4 2001 Page 2 001346 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-308 Page 122 Peak Area Report Sam ple Nam e: 1095JGROUP4-F1 Notebook Rateranee: PABX-BBA-1-047 Date Acquired: 1/3/01 7:33 O p erato r. Data FHe Nam e: 0471061.0 Data Fila Path: O:\MSD_DATA\PAB\PABX-BBA\1-047-1\ tnatn/m ent M ethod Nam e: PABX_BBA tiatn im en t Name: MS7 hatnim ent Vial N um ber 61 Comoound t U 2) Comoound Nomo MVL MVL-4 Rtendon Tim e fM ln.l 7.02 7.0 jon 131 135 Peak Area 241386 2084 Peak Area Ratio 115.8E+0 M* - The te n * *m * In thle column Ind u that menuel In ag rerlan wee pirto im ed on e m compound. Print Date ri*--.' Time: 1/18T . 15:05 age 1 001347 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-309 Page 123 Quantitation Report IQT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PABNPABX-BBA\ 1-047-l\0471061.D Vial: 61 3 Jan 2001 7:33 Operator: 10953-GROUP4-F1 Inst : MS7 Integration Pararne : RTEINT.P Lant Method : D:\HPCHEM\1\METH0DS\PA8X_BBA.M (RTE Integrator) M V L ____________________ ___________ Amount :_____ 0.00 " 'ion':'131 (7.016) !Resp: 241386 0471 PC 1 .D PAP'< BBA.M Thu Jar IB 15:ir :\4 2001 p> : 2 001348 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-310 Page 124 Peak Area Report Sample Name: 109S4-GROUP4-F1 Notebook R aleienee: PABX-BBA-1-047 Data Acquired: 1/3/01 7:53 O perator Data FHe Name: 0471062.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Inatrum ent Method Name: PABX.BBA kiatrum ent Name: MS7 tnatmment Vial Num ber. 62 Com ooim d f U 2) Comoountf Name MVl MVLhM Retention Tim e fM ln.) 7.02 7.04 Ion 131 135 Peak Area IT 226425 1844 Peek Area Ratio 122.8E+0 M* mThe Iettar V In Oita column Indiearee met manual Integration m s parlormad on that compound. Print Date and Tene: 1/18/01 15:05 Page 1 001349 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-311 Page 125 uuinucacion Report tui nevieweu/ Data File Acq On Sample Mise D:\MSD DATA\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\1\METH0DS\PABX_BBA.M (RTE Integrator) MVL._____ "ion: 3 "(T. O'bT :Resp: 226425 _____________________ A m o u n tj_________0 0 0 _______________________________________________ _________ *i i 04 V ' 362. D PABX BBA.M Thu Jan 18 15:0 34 2001 age 2 001350 Primedica-Worcester Project Number. PABX-BBA 418-018:PAGE 1-312 Page 126 Peak Area Report Sam ple Name: 10943-GROUP4-F0 Notebook Reference: PABX-BBA-1-047 Data Acquired: 1/3/01 8 :t2 O p erator: Data File Name: 0471063.D Date File Path: D:\MSD_0ATAtPAB\PABX-BBA\1-047-1\ Instrum ent M ethod Name: PABX.BBA Instrum ent Name: MS7 Instrum ent Vial Number: S3 Comoound i) 2) Comoound Nomo MVL MVL-4 Retention Time (M in.l 7.02 7.04 Ion 131 135 p m .K.Am | 1753 1939 e n k iM .R a 4 f 904.1E-3 It* The letter *m* In this cdumn Indicates bier manual imaprebon area pertenned en thet compound. Print Date and Time: 1/18/0n 15:05 Page 001351 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-313 Page 127 S/uincu.auuu nepuiu Data Pile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471063.D Vial: 63 3 Jan 2001 8:12 Operator: 10943-GROUP4-F0 Inst : KS7 )Integration Parame: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator M V X . _______________ ________________ Amount ;.... 0.00 ' `on:'"l3 (7.016) * .... * 0471063.D PABXBBA.M Thu Jan '8 15:05:53 2001 Fv,t 2 001352 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-314 Page )?8 Peak Area Report Sample Name: 10948-GROUP4-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 8:32 O perator Date File Hama: 0471064.0 Data File Path: D:\MSD_DATA\PAB\PABX-BBA11-047-1\ Instrum ent Method Name: PABX.BBA bietram ent Nam e: MS7 kiatnim ant Vial N um ber 64 Comoound 1) 2) Compound Name MVL M VL-04 Retention Time (M in.) 7.02 7.04 te a 131 135 Peak Area ( 2188 1996 Peak Area Ratio 109.6E-2 h r The M a r *m * In M * column kuNcaac Oiat manual kia yaU un ama parlanim il an Star compound. Pnr. 'ale and Tmc: 1/18/01 1f:( Page 1 001353 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-315 Page 129 yuantitacion neport tui evimeui Data File : D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471064 .D Viali 4 Acq On 3 Jan 2001 8:32 Operatori Sample : 10948-GROUP4-F0 Inst : MS7 Mise Integration Paratasi RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVXj_______ ______ __________________ Amount :______ 0.00 f Ioni 131' (7".017) 0471064.D -ABXBBA Th>Jan 18 :,:06:13 . ' vi Page 7 001354 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-316 Page 130 Peak Area Report Sample Name: 10952-GROUP4-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 8:52 O p erator Data File Name: 0471065.D D aU FHe Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX.BBA kratrum ent Neme: MS7 kiatnim ent Vial N um ber 65 Comoound 4 0 2) Comoound Name MVL MVL-4 Retention Tim e fM in.l 7.01 7.04 Jen 131 135 PeakArea M* 1778 2817 Peak Area Ratio 679.4E-3 M* - The tetter "m* in eke column indieelea diet manual Integration ama performed an met compound. P m t Date a:.d Time: 1/18/0' 15:06 -ge 1 001355 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-317 Page 131 Data File Acq On Sample Mise uuancitatxou itepuiw \vi wsv*oeu/ P:\MSD DATA\PAB\PABX-BHA\1-047-1\0471065.D Vial: 65 3 Jan 2001 8:52 Operator: 10952-GROUP4-F0 Inst : MS7 MVL __ ___ ________ ________________ Amount :______ 0 00 ! Ion :~131 (7. Oisl 0471CS.D PAP'". BBA.M Thu Jen 18 15:'u.32 2001 Pf . 2 001356 Primedica Worcester Project Number: PABX-BBA 418-018:PAGE 1-318 Page 132 Peak Area Report Sample Name: 10955-GROUP4-F0 Notebook Roteranee: PABX-BBA-1-047 Date Acqutrad: 1/3/01 9:11 O perator Date FUe Name: 0471066.D Date FNc Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1V Inatnim ent Method Name: PABX_BBA Inatnim ent Name: MS7 Inatnim ent Vial Number. 66 Com pound 1) 2) Comoound Name MVL MVL-d4 Retention Tim e (M in.) 7.02 7.04 i2 a 131 135 Peak Area M* 3322 2412 Peak A iea Ratio 137.7E-2 M* The Mear *m* m eus column indcalos teal manual integration m e pwtarmad on teal compound. Pnnt Dale and Time: 1/18/01 15.06 Pag.- 1 001357 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-319 Page 133 DaCa File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471066.D Vial: 66 3 Jan 2001 9:11 Operator: 10955-GROUP4-F0 Inst : MS7 MVL ______________ _______ ____ ____ Amount:______ 0_.00 ' ion: 31 (7.6*18)" . >71066.1' PABX BBA .M <u Jan 18 75:06:5.? -001 Page 001358 Primedica-Worcester Project Number. PABX-BBA 418-018:PAGE 1-320 Page 134 Peak Area Report Sample Name: 10956-GROUP4-F0 Notebook Reference: PABX-88A-1-047 Date Acquired: 1/3A31 9:31 O perator. Date File Name: 0471067.D Date File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX.BBA butrum enl Nam e: MS7 hwtrument V ial N um ber. 67 Comoound * 1) 2) Com oound Name MVL M VI-4 Retention Tima /M in.) 7.01 7.04 jOQ 131 135 Peek Area M* 357223 2339 Peak Area Ratio 152.7E+0 M* Tbs tedar "m" in this cokimn Indicates that manual tnteysthm was performad on that compound. Prmt Date and Time: V i b /01 15:07 Pag. 001359 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-321 Page 135 W uaiit-.L L ai.A U ti R e p til l tWL R e v i e i e u t Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471067.D Vial: 7 3 Jan 2001 9:31 Operator: 10956-GROOP4-FO Inst : MS7 Integration Parants: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) M V L _____________________ _____________ Amount :______ 0.00 ; Ion: 131 (7.014) . Resp: 357223 r.'.71067.1. PABX BBA.M Tnu Jan 18 15:07:11 iOOl Page . 001360 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-32? Page 136 Peak Area Report Sam ple Name: 10944-GROUP4-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 173/01 9:51 O perator Data File Name: 0471068.D Data File Pat: O:\MSO DATA\PAB\PABX-BBA\1-047-1\ Instrum ent M ethod Name: PABX BBA Inatnim ent Name: MS7 Instrum ent Viel Num ber 68 ComBOund # U 2) Comnound Name MVL MVL-d4 Retention Tim e flW n.l 7.02 7.04 Is s 131 135 Peak Aree JT 2815 2414 Peak Area Ratto 116.6E-2 ht* b The letter-oT In this column Indlr otot that menuet Integration nee performed on thet compound. Print Dale and Time: 1/1B/01 15:07 Page 1 001361 Priinedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-323 Page 137 V*"41 Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471068.D Vial: 6 3 Jan 2001 9:51 Operator: 10944-GROUP4-F0 Inst : MS7 Integration Parama: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0771068.D '.BX BBA.h Thu rn 18 15 07:31 2001 Page 2 001362 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-324 Page 138 Peak Area Report Sample Nam e: 11029-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 10:11 O perator Data R ie Nam e: 0471069.D Data R le Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ kiatrum ent M ethod Nam e: PABX.BBA In itn im en t Name: MS7 Inetrvm ent Vial N um ber 69 Comoound t 1) 2) Comoound Name MVL M VL44 R etention Tim efM irU 7.02 7.04 Ion 131 135 Peak Aree M* 781 1833 Peak Ama Rado 426.1E-3 M* The M a r *nr in UUa cokjmn M ic e a e e<at ewnuel Inagretlen wee performed en UM compound. Print Date and Time: 1/1 SOI 15:07 Page 1 001363 Primedica-Worccster Project Number: PABX-BBA 418-018:PAGE 1-325 Page 139 wuantitation Report lUi itevieweaj Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-07-l\0<71069.D Vial: 69 3 Jan 2001 10:11 11029-GROUP11-FO Operator Inst MS7 J Integration Params: RTEINT.P bant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M IRTE Integrator) M V L ___ _____________________ ______ Amount :______ 0.00 IonT"131 17.016) Resp : 781 0471069. '> PABX E' ' M rhu Jan ?r .5:07:* 2001 Page 001364 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-326 Page 140 Peak Area Report Sample Nam e: 11031-GROUP11-F0 Notebook Reference: PABX-BBA-1-047 D a li A cquired: 1/31 10:31 O perator. Data R ia Nam e: 0471070.D Data R ia Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Inatnim ent Method Nam e: PABXJ3BA Inatnim ent Name: MS7 kiatrum ant Vial Num ber. 70 Comcound f 1) 2) Com oound Name MVL MVL-4 Retention Tim e (M in.) 7.02 7.04 le a 131 135 Peak Amo M* 710 1713 Peak Area Ratio 4 14 .5E -3 M*Theletter*m* InthiscoturniInillisun matmanualIntegrationneeparformoOendialcompound. Print Date and Tme: , i /01 15:08 Page 01365 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-327 P age 141 Quantitation Report iur Kevieweoj Data File Acq On Sample Mise D-\MSD DATA\PAB\PABX-BBA\ 1-047-1X0471070 .D Vial: 70 3 Jan 2001 10:31 Operator: 11031-GROUP11-F0 Inst : MS7 Integration Parama: RTEWT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.H (RTE Integrator) 047107C 0 PABX RBA.M Thu Ji , lB 15:08- 2001 Pag.. 2 001366 Primedica Worcester Project Number: PABX-BBA 418-018:PAGE 1-328 Page 142 Peak Area Report Sam ple Name: 2 X QC MIO Notebook Retetenca: PABX-BBA-1-047 Date Acquired: 1/3101 1050 O p erato r: Data File Name: 0471013.D Date File Path: D:\MSD_DATA\PAffPABX-BBA\1 -047-1\ biatnim ent Method Name: PABX_BBA kutnim ant Name: MS7 Inatnim ent V M Num ber. 13 Comoound 1) 2) Comoound Name MVL MVL-4 R etention Tim e (M in.) 7.02 7.04 Ion 131 135 Peak Ama M* 737 15S4 Peak Area Ratio 4 6 5 J E -3 M ' The letter *m* In Ulte column Indicate that manual Intepratlorneaa partermao an that com pcw L I rttit Date and True: 1/1B/01 48 Pr' lt 1 001367 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-329 Page 143 uuantitaticm nepuiu tu* rc*c c u / Data File : D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471013 .D Vial: 13 Acq On 3 Jan 2001 10:50 Operator: Sample 2 X QC MID Inst : MS7 Mise MVL : Ion: 131 ("7.015) :Reap: 737 _ __ Amount:_____ 0.00 L '471013.'.' PABX p: M Tin Jan 1 34:48:48 2001 Page >. 001368 Primedica- Worcester Project Number: PABX-BBA 418-018:PAGE 1-330 Page 144 Peak Area Report Sam ple N a rra : 11032-GROUP11-F0 N otebook R ataranca: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 11:10 O p e ra to r Data FHaNam e: 0471071 JO Data nie P ath: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1V In stru m e n t M ethod Nam e: PABX.BBA btetm m ent N am e: MS7 kiatrum ent V ial N um b e r 71 Comoound * 1) 2) C om oound Name MVL MVL-4 R etention T im * /M in.) 7.02 7.04 Ion 131 135 Peak Area 285 1766 Peak Area R atio 161.4E-3 M* MThe latter "m" In mis column indicates mat manual Integration was performed an that compound. P m t Dale and Time: 1/18/01 15:08 Pagr 001369 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-331 P age 145 yuancitacion Report itfi nevieweaj Data Pile Acq On Sample Mise D:\MSD DATA\PAB\PABX*BBA\1- 047 -1\0471071.D Vial: 71 3 Jtn 2001 11:10 Operator: 11032-GR0UP11-P0 Inst : MS7 Integration Parana : RTEINT.P ant Method : D:\HPCHEM\l\KETHODS\PABX_BBA.M (RTE Integrator) 0471071. '> PAfiX EBA.M ^hu Jan lc- .5 :0B:30 101 Page 001370 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-332 Page 146 Peak Area Report Sam ple Nam e: 11033-GROUP11-FO N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 11.30 O p e ra lo r. Data R ie Name: 0471072.D Data R ie P ath: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ In stru m e n t M ethod Nam e: PABX.BBA ktstrum e n t Nam e: MS7 In a tn im a flt V ia l N um ber. 72 Comoound # 1) 2) C om oound Name M Vl MVL-4 R etention Tim e fM in.) 7.01 7.04 is a 131 135 P w hA rea 481 2422 Peak Area R atio 198.6E-3 r mThe letter *m* tn thle cotumn indicete that manuel In tesatien a s performed an that compound. Pnrt Dale and Time: 1/18/01 15:08 pa. 1 001371 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-333 Page 147 u w fve<rxeeuf 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-GROUP11-F0 Inst : MS7 Integration Params: RTEXKT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BHA.M (RTE Integrator) KVL___________________ ______________ Amount :_____ 0.00 ; Ion:* 131 (7.012) 0*71072.D PAEX BBA. Th1 Jan 18 06:49 2*"l Page 2 001372 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-334 Page 148 Peak Area Report S am ple Nam e: 11O34-GROUP11-F0 N oteb o o k R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 11:50 O p e ra to r D ata F ile Nam e: 0471073.D D ata F ile P ath: D:\MSD_DATA\PAB\PABX-8BA\1-047-1\ In stru m e n t M ethod Nam e: PABX_BBA In stru m e n t Nam e: MS7 kia tn im e n t V ia l N u m b e r 73 Com oound t 1) 2) II* The lenir C om oound Name MVL M V l-44 R etention Tim e W in .) 7.01 7.04 te a 131 135 Peak A rsa II* 12941 2613 Peek Am a R atio 495.3E-2 1In this column Indicate manual liHisllon wa pertnrmaJ on Siat compound. rm t Date and Time: 1/18/Q1 15:09 ?age 1 001373 Primedica-Worcester Project Number: PABX-BBA 418-018.PAGE 1-335 Page 149 Data File Acq On Sample Mise uuantitation Report itri nevieweaj D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471073.D Vial: 73 3 Jan 2001 11:50 Operator: 11034-GROUP11-F0 Inst : MS7 M V L _____________ _________ __ _ Amount ;______0.00 ; `ion: 131 (7.013) " ~ ..... . 0471073.J PABX BBA.M ''.i Jan li 15:09:O' 7001 Page ' 001374 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 O perator Data FRe Name: 0471074.D Data FHe Path: O:\MSD_DATA\PAB\PABX-BBAVl-047-1\ Instrum ent Method Name: PABX.BBA Instrum ent Name: MS7 Instrum ent Vial Number. 74 Compound 1) 2) Comoound Name MVL MVL*d4 Retention Time iM In.l 7.02 7.04 ton 131 135 Peak Area M* 647 2446 Peak Ama Ratto 264.3E-3 M* The le tT *m* In this column indicates that manual Inla y aMcnnee performed en that compound. Print Date ir.ri Tme: 1/1B/01 15:09 "age 1 001375 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-337 Page 151 V ju a n tiu a ix im jiepuxi. vvx ivcv c e u i Data File Acq On Sample Mise D:\MSDDATA\FAB\PABX-BBA\l-047-l\0471074.D Vial: 74 3 Jan 2001 12:10 Operator: 11035-GROUP11-P0 Inst : MS7 Integration Parane : RTEIMT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 04r. 074.D PABXBBA.M Thu J.\n 18 l r '79:29 2001 2 001376 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-338 Page 152 Peak Area Report Sam ple Nam e: 11036-GROUP11-F0 N otebook R eferance: PABX-BBA-1-047 Date A cq u ira d : 1 /3/01 12:30 O perator: Data F ile N am e: 047107S.D Date F ile P ath: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1N In e tn im e n t M ethod N am e: PABX.BBA In e tn im en t Nam e: MS7 In e tn im en t V ia l N um b e r 75 Comoound i) 2) C om oound Name MVL M VL44 R etention Tim e flUlln.t 7.02 7.04 {on 131 135 Peak Area M* 526 1566 Peak Area R atio 335.9E-3 M* * The letter -m" In pile column indlcalee that manual Integration irne performed on diet compound. P ri: Date and f. u . 1/18/01 15:09 Pevc 1 001377 Priraedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-339 Page 153 uuantitacion Report \ui Reviewed,/ Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471075 .D Vial: 7S 3 Jan 2001 12:30 Operator: 11036-GROUPll-FO Inet : MS7 Integration Parama: RTEINT.P ant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) 0471075 !> PABX BBA.M '"hu Jan iC 15:09: 2001 Pagr 001378 Primedica-Worcester Project Number: PABX-BBA 418-018.-PAGE 1-340 Page 154 Peak Area Report Sam ple Nam e: 11037-GROUP11-F0 N otebook R eference: PABX-BBA-1-047 Date A cquired: 1/3/0112:50 O p e ra to r D ata FHe Nam e: 0471076.D Data F ile P ath: D:\M SD_DATA\PAB\PABX-BBA\1-047-1\ b istru m e n t M ethod Nam e: PABX_BBA tn a tn rm e nt Name: MS7 tn a tru m e nt V ia l N um ber 76 Comoound t 1) 2) C om oound Name MVL MVL-4 R etention Tim e (M in.) 7.02 7.04 Isa 131 135 Peak Area M* 542 2016 Peak Area R atio 268.BE-3 M* th e letter *m* In Pile column Indicates that manuel Integration ree performed on tuet compound. Print Date and Time: 1/18/01 15:10 ge 1 001379 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-341 Page 155 Uuantitation Keport tux Kevievieai Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX*BBA\1-047-1\04 71076 .D Vial: 76 3 Jan 2001 12:50 Operator: 11037-GROUP11-F0 Inst : MS7 Integration Params: RTEINT.P Jiant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) 0471076.D PABX BBA.M Thu Jan 18 15:10:'' 2001 Par. 2 001380 Primedica-Worccster Project Number: PABX-BBA 418-018:PAGE 1-342 Page 156 Peak Area Report Sam ple Nam e: 11038-GROUP11-F0 N otebook R eference: PABX-6BA-1-04? Date A cq u ire d : 1/3/01 13:10 O perator. Data F ile Nam e: 0471077.D Data File P ath: D:\M SD_DATA\PABiPABX-BBA\1-047-1\ Instrum ent M ethod Nom e: PABX.BBA kia tn u n e n t Nam e: MS? M etrum ent V ia l N um b e r 77 Comoound * 1) 2) C om oound Name MVL M V L-d4 R etention Tim e (M in.i 7.02 7.04 131 135 Peak Area f i : 602 2007 300.0E-3 M' The HOST 'i n ' m m u column InOIrs tes that mereiai ensQuUon wes porTormad on a m compound. Print Date and Vne:1/1B/01 IS : i0 iWie 1 0013S1 Primedica-Worccstcr Project Number: PABX-BBA 418-018:PAGE 1-343 Page 157 uuantitation iceporc iui aevieweuj Daca File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\ 0471077 .D Vial: 77 3 Jan 2001 13:10 OperaCor: 11038-GROP11-F0 Inst : MS7 Integration Parama: R T E I N T .P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) 047107 ;.D PABX B B A .M iu Jan 18 15:10:23 '001 Pag 001382 Primedica-Worcester Project Number: PABX-BB 418-018:PAGE 1-344 Page 158 Peak Area Report Sam ple Nam e: 11029-GROUP11-F1 N oteb o o k R ele ie n ce : PABX-BBA-1-047 Date A cq u ire d : 1/3/01 13:30 O p e ra to r Data FHe Nam e: 0471078.D Data FHe P ath: D:\M SD_0ATA\PAB\PABX-BBA\1-047-1\ b ie tn im e n t M ethod Nam e: PABX.BBA In s tn im e fit Nam e: MS7 k u tn im e n t V ial N um b e r 78 Com oound # 1) 2) Com oound Name MVL M VL44 R etention Tim e (M in.) 7.02 7.04 le c 131 135 Peak Aree M` 189 1446 Peak Area R atio 130.5E-3 M* The tener "m* In this column Indicate die t manuel kitey a lkin wee partormea on that compound. Poni Dale : <1 Time: 1/18/01 15:10 Page 1 0013S3 Primedica-Worcester Project Number PABX-BBA 418-018:PAGE 1-345 Page 159 Uuantitation Report iui itevieweo; Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471078.D Vial: 78 3 Jan 2001 13:30 Operator: 11029-GROUP11-F1 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEMU\METHODS\PABX_BBA.M (RTE Integrator) MVL_________________ Ion:' 131 (7.06 Reap: 189 __ _Amount:____ 0.00_______________ ______ ___________________ ........ . "".... -- - _ I ! 0471078.D PA3;. Thu Jan 18 15:M:. :8 2001 P.-.-I. 2 0013S4 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-346 Page 160 Peak Area Report Sam ple Nam e: 11031-GROUP11-F1 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d : 1/3/01 13:49 O p e ra to r Data F ile Nam e: 0471079.D Data FHe P a tti: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ kiatrum e n t M ethod Nam e: PABX.BBA In a tn im e n t Nam e: MS7 In a tn im e n t V ia l N um b e r 79 Comoound U 2) Com oound Nam* MVL MVL-44 R eten tio n T im e (Min.) 7.02 7.04 Ion 131 135 Pepk Arpa 146 1617 Peak Area Ratio 902.9E-4 M* The M a r'm * In ma column Indicaas that manual integration ana performed on that compound. Port Date and Tune: 1/18/01 ibriO P:-.. 1 001385 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-347 Page 161 Quantitation Report lUT Reviewed) Data Pile Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471079.D Vial: 79 3 Jan 2001 13:49 Operator: 11031-GR0UP11-Fl Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) MVL _______ _____ _____________ Amount :______ 0.00 T Ion:"3" (7.019)" jResp: 146 0 4 W 0 7 9 . D `'ABX BBA Th.' Jan 18 15:11:08 2001 -age 2 001386 Primedica-Worcester Project Number PABX-BBA 418-018:PAGE 1-348 Page 162 Peak Area Report Sam ple Nam e: 11033-GROUP11 *F1 N otebook R eference: PABX-BBA-1-047 Dale A cq u ire d : 1/3/01 14:09 O perator. Data F ile Nam e: 0471080.D Data F ile P ath: D:\MSD_DATA\PABNPABX-BBA\1-047-1\ In strum ent M ethod Nam e: PABX.BBA kiatm m en t Nam e: MS7 kietram e n t V ial N um b e r 80 Comoound # i) 2) C om oound Name MVL M VL-04 R etention Tim e (M in.) 7.02 7.04 ta n 131 135 Peak Area M* 293 1957 Peak Area R atio 149.7E-3 M* The tetter "m* In this column Indicates that manual Inte y a tlun ame pertormed on mat compound. Pi ' lateandTir. : i/18/01 l f .1 Paur 1 001387 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-349 Page 163 Data File Acq On Sanale Mise Quantitation Report tur Revieweai D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471080 .D Vial: 80 3 Jan 2001 14:09 Operator: 11033-GROUP11-F1 Inst : MS7 MVL ________ __________________________ Amount:______0.00 r io H :"T 3 i !7ro i6 ) 0 4 7 1080.D -'iX BBA.H Thu Jan 18 15:11:28 2001 je 2 001388 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-350 Page 164 Peak Area Report Sam ple Nam e: 11036-GROUP11-F1 N otebook R eference: PABX-BBA-1-047 Date A cq u ire d ; 1 /3/0 1 14:29 O p e ra to r Data R ie Nam e: 0471061.0 Data R ie P ath: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1 \ In stn im e n l M ethod Nam e: PABX.BBA In stru m e n t Nam e: MS7 In stn im e n t V ial N um b e r 81 Compound i n 2) C om nound Neme MVL M VL-04 R etention Tim e (M in.) 7.02 7.04 ton 131 135 Peek Aree M* 260 1943 Peak A n a R atio 133BE-3 M* * The tetter "m* In this column matetoss met manual tmogrstton was performed on that compound. Prin :Jate and Time: 718/01 15:M Pc' Pay. i 001369 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\1 -047-1 \0471081 .D Vial: 81 3 Jan 2001 14:29 Operator: 11036-GROP11-F1 Inst : MS7 M V L _ ______ ___ ____ __________________ Amount " 'Ion: 131 CT.bl) Resp: 260 ___ 0 .0-9- 0 7 1 0 8 1 . 1 . "'ABX BBA.H Tl.v Jan IB 15:11:47 2001 Page / O O lfO 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\l-062-l\062l057.D Vial: 57 IB Oct 2000 8:04 Operator: MAN 6 x Control Plasma Inst : MS7 MS Integration Parana: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) fndanos ! 2000! 1000 TIC: 0621057.0 f; nil sool IrTM- 6.10 6.20 6.30 6 *0 6.50 6.60 670 6.60 6.90... 7.00 7.10 7.20 7.30 7.40 7.50 7.80 7.70 7,60 7.90 M V L .... ...... .......... Amount: 0.00 .lori31 (7.134) Resp: 503 .........../ v . . . . . . . . . ____________ _____________________ ! --n -- 1 i y --- i - n-- i` --r , -- -- i 1 " i 630 6.30.....6 *0 650 6.60 6.70 6.80 6.90 7 0 0 7.10 7,20 7.30 -d 4 _____ __________________________ ' Amount: f cIon: 135 (7.162) !Resp: 4373 ' T ' - v - . . r .......................... i -- -- ............. ...... 7 *0 7.50 7.60 7.70 7 50 7.90 0.00 1... . ! 6.20 6.30 6 *0 6.50 6.60 6.70 6.60 6.90 7 00 7.10 720 7.30 7.*0 7.50 7.60 7.70 7.80 7.90 0621057.D PABX_BVA.M Wed Oct 18 12:58:08 2000 Page 2 001391 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-168 Page 167 Peak Area Report Sam pta N am e: 2 x B lank W a t N otebook R ateranco: PABX-BVA-1-0S2 D ate A cq u ire d : 10/18/2000 B.23 O p e r a to r MAN Data F ila N a n o : 0621OSB.O Data Fite P ath: D:\M SO_DATA\PAB\PABX-SVA\1-062-1\ Inabum ant Method N a n a : PABX_BVA In ttru m e n t N a n a : MS7 In a tru m a nt V ia l N um bar: 58 Com pound 1) 2) C om pound Nama MVL M V L-d4 R etention TJimlMIft) 0.00 0.00 Is a 131 135 Ea*ftrm tc 0 0 P o a k A ia a fia lto CNV/0! M* The la tu fin " in mia coturno Indicata teat manual Inteptt Boioae pertamiod an a p t compound. Print Data and Tana: 10 n 8/2000 12:58 Page 1 001392 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-169 Page 168 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621058.D Vial: 58 18 0ct~2000 8:23 Operator: MAN 2 x Blank. Water Inst : MS7 MS Integration Pararne: RTBINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTB Integrator) jfcbundanea TIC:0621066.0 3001 ! iI 290| iK A 500- X - ' A ___ V. 6.10 6.20 6.30 6.40 6.50 6.60 6.70 6,60 660 TOO 7.10 T.2D 7.30 7.40 7.50 7 60 7.70 7.60 70 M V L ... ... ........ ' ' _ A'mount- :- N-o-t -Found I Ion: 13 (.) I^ M A J 6.20 6.30 6.40 6.60 6.60 6.70 6.80 6.90 7.00.7,10 720 7.30 7 40_ 7.50 7.60 7.70 760 790 -d4 Amount: Noi:'"Found' Ion: 135 (0.000) i /\ 6.20 630 640 650 6.60 6-70 6.60 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 760 790 0 6 2 1 0 5 8 . D PABX BVA.M Wed O c t 18 1 2 : 5 8 : 2 8 2 0 0 0 Page 2 001393 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-170 Page 169 Peak Area Report Sample Naim: 2x Commi Water Notebook Reterenoe: PABX-BVA-1-082 Date Acquired: 10/18/2000 8:43 Operator:MAN Data FHe Name: 0621059.D InetnimantNeme: MS7 Data FHe Path:D:\MSD_OATA\PABtf>ABX-8VAV1-062-1V InstrumentVialNumber: S9 Instrument Method Name: PABXJ3VA Compound 1) 2) Compound Name MVL MVL-04 Retention IHntlMinJ 0.00 7.16 ten 131 135 Peak Aree { 0 5369 P n k AraaJtatio ooo.oe-3 M* The letter *W in this column indicetee the* inenuei integration ewe performed on diet Pnnt Dote and Time: 1C/1SO0O 12.5 Peg 1 001394 Primedica-Worcester Project number: PABX-BVA 418-018: PAGE 1-171 Page 170 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621059.D Vial: 59 18 Oct-2000 8:43 Operator: MAN 2 x Control Hater Inst : MS7 MS Integration Params: RTEINT.P Quant Method : D:\HPCHEM\1\METH0DS\PABX_BVA.M (RTE Integrator) KEnSnT TIC:0621Q59.D 2000- .1000 MVL Ion: 131 (0.000) ii Amount: Not Found A .-- \.^\ l/ W A "W J\ V -- ,vyV A -/A -- i 6.20 630 6.40 650 6.60 6.70 6.00 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.80 750 M V L-d4 Amount : 0.00 Ion: 135 (7.163) Resp: 5369 !i ; v. ! 6.20 6.30 6.40 6.50 660 670 8.80 690 700 7.10 7.20 7.30 7 40 750 7.60 7.70 7.80 7.90 0 6 2 1 0 5 9 .D PABX_BVA.M Wed O c t 18 1 2 : 5 8 : 4 7 2 0 0 0 Page 2 001395 Priniedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-172 P a g e 171 Peak Area Report Sample Name: 2 x STD-1 Water Notebook Reference: PABX-BVA-1-062 Data Acquired: 10/18/2000 9 02 O perator. MAN Data File Name: 0621060.D Data File Path: D:\MSD_0ATA\PAB\PABX-BVA\1-0e2-1\ Instrum ent Method Name: PABX_BVA Instrum ent Name: MS7 Instrum ent VW Num ber. 60 Com oound# 1) 2) Comoound Name MVL MVC-d4 RatonCon Tim a (M in.) 7.13 7.16 lea 131 135 PeakA rae M487 8305 Paak Area Ratio 772.4E-4 H r The lattar "m" rn this column indicates that manual integration Wat performed on that CMK1mairi. Print Date and Tim *: 10/18/2000 12:58 Page 1 001396 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-173 Page 172 Quantitation Report (QT Reviewed) Data File Acq On Sample M6C D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621060.D Vial: 60 18 Oct 2000 9:02 Operator: MAN 2 x STD-1 Hater Inst : MS7 MS integration Params: RTEINT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) 0 6 2 1 0 6 0 .D PABX_BVA.M Wed O c t 18 1 2 : 5 9 : 0 6 2 0 0 0 Page 2 001397 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-174 Page 173 Peak Area Report Sample Kama: 2 x STD-2 W ater Notebook Reference: PABX-8VA-1-062 Date Acquired: 10/18/20008:22 Operator: MAN Data File Name: 0621061.D Instrum ent Name: MS7 Data File Path: 0:W S0_0ATA\PAB\PA8X-BVA\1-062-1V Instrum ent Vial Number: 61 In itn im e n t Method Name: PABX_BVA Com pound 1) 2) Com pound Name M Vl MVL-64 Retention Tim e IM In.l 7.14 7.16 bn 131 135 Pw a th m: 781 m 5494 Peak A lea Ratio 142.2E-3 M* " The letter "m" in this calum i indicates that manual imegfetron aies performed cn that compound. P rint Date and Time: 10/15/200012:59 Pag 1 001398 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 Pararne: RTEINT.P Quant Method : D:\HPCHBM\l\METHODS\PABX_BVA.M (RTE Integrator) iteundanca !I TIC:0621051.0 20001 I\ /v |Tinto-> 6.10 6-2D 6.30 6.40 6.80 60 6.70 6.B0_ 6.90 7.00 7.10 7.70 7.30 740 7.50 7.00 7.70 7.80 ISO MVL ....... Amount : 0.00 .. .... I Ion: 131 (7.137) iResp: 781 ! . [\ ! I\ 'T -- 1 6.7Z0C 6.30 6.40 6.50 6.60 670 660 630 7.00 7,10 7.20 7.30 7.40 7.S0 7.60 7.70 7,60 7.80 -d4~ ... ...................... Amount: 0.00_ 9 F tI,onV 135 '('7.160)' !Resp: 5494 I fl I 620 6.30 640 6.SO 6.60 6.70 6.60 630 7. 710 770 730 740 750 7.60 7.70 7.M 7.90 0 6 2 1 0 6 1 . D PABX BVA.M Wed O c t 18 1 2 : 5 9 : 2 6 2 0 0 0 Page 2 0013S9 Primedica-Worcester Project number PABX-BVA 418-018:PAG EM 76 Page 175 Peak Area Report Sam ple Nam: 2 x STD-3 Water Notebook Ratbfanoe: PABX-BVA-1-062 Data Acquired: 1W16/3000 41 Operator: MAN Data File Name: 06210B2.D Data F it* Path: D:\MSD_DATA1PAB1PABX-BVA\1-062-1\ Instram ant M ethod Nam e: PA8X_BVA biatnim entNam e: MS7 Nistrum ant Vial Num bar. 3 Com pound 1) 2) S sm fiS unA tim t MVL MVL-04 R atantion Tim iMtn!) 7.14 7.16 is o 131 135 E M tlftfW 1C 1305 6271 Peak Ama Ratio 208.16-3 P rint Date and Tim e 10/18/2000 12:50 Page 1 001400 Primedica-Worcestcr 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:\HPCHRM\1\METH0DS\PABX_BVA.M (RTE Integrator) Abundanea | 3000i i i! 2000 lOOOj TIC:0621082.0 A ft i \\ !\ T rv'--'T' 0 . 0 06,10 6:20 630 640 6.80 660 6.70 6.60 690 7.00 7.10 730 730 740 7.60 7.60 7.70 7.60 7.90 MVL Amount : Ion: 131 (7.135) Resp: 1305 i i i i j A 11 6.20 6,30 6.40 630 6.60 6.70 6.60 6.80 700 7.10 7,20 7.30 7.40 7-.56C0 7.80 770 7.60 7.90 -d4 Amount : 0.00 Ion: 135 (7.162) Resp: 6271 620 6.30 640 650 6.60 6.70 660 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7,60 700 0 6 2 1 0 6 2 . D PABX BVA.M Wed O c t 18 1 2 : 5 9 : 4 7 2 0 0 0 Page 2 001401 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-178 Page 177 Peak Area Report Sample Nam e: 2 x STD-4 W e t* Notebook Reference: PABX-BVA-1-062 Date Acquired: 10/18/200010.01 Operator: MAN Date File Nam e: 0821063.D Date FHe Path: D:\M50_QATA\PAaPABX-8VAVI-062-1V Instrum ent Method Name: PABX.BVA Instrum ent Name: MS7 Instnim ant V ial Number: 83 Compound f 1) 2) Pppipptprt W m MVL MVL*d4 Retention Ijfflp J U iQ j 7.13 7.18 1SD 131 135 Peak Area JT 1817 m 5685 Peak Area R ails 337.2E-3 HT > The letter "m" in mis column indicelee thet manuel imepratien w e* performed on that compound P rint Date ana Time: 10/18/2000 12:59 Page l 001402 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-179 Page 178 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621063.D Vial: G3 18 Oct-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 (RTE Integrator) *hund^o 3000: | aoooi1 1000; TIC.0621063X1 A I! j 1 \ ! ;l /\ !Tvt*-> 6 1 0 6 2 0 6.30 640 6.90 6.60 6 70 6.80 6.90 7.00 710 7.20 730 7.40 7.50 7.60 7.7D 7.80 790 MVL Amount: o.oo Ion: 31 (7.134) Resp: 1917 l\ ! ! '! ;\ 6.20 6.30 640 6.50.6.60.6,70 /L-d4 Ion: 135 (7.161) Resp: 5685 6.60 6.90 7.0 710 720 7 Amount: .......... " ' A 740 7.50 7.60 7.70 7.60 7.90 0.00 .. .... . 6.20 630 6.40 6.60 6.60 6.70 6.60 6.90 7.00 7.10 7.20 7.30 740 750 7.60 7.70 760 790 9 0621063.D PABX BVA.M Wed Oct 18 13:00:06 2000 Page 2 001403 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-180 Page 179 Peak Area Report Sample Name: 2 x STD-5 W a t Notebook Referanca: PABX-BVA-1-062 Data Acquired: 10/18/2000 10-20 Operator: MAN Dato FHa Name: 0621064.D Oato FHe Path: D:\MSO_OATA\PAB\PABX-BVA\1-062-1\ Inatrum ent Method Name: PABX_BVA Instnim ent Name: MS7 Instrum ent Via< Number: 64 C9m w w n# 1) 2) Com pound Name MVL MVL-d4 R tta rtio fi Tim (MhU 7.13 7.16 lo g 131 135 E e& A m tc 2410 4094 Peak Area Rado 588.7E-3 t r Tha la ti "laT* la this column indicata* H ut ta m a l Integrat ion araa portocmod on that compound. Print Data and Time: 10/16/2000 13:00 Page 1 001404 Primedica-Worcester Project number: PABX-BVA 418-018:PA G E 1-181 Page 180 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD_DATA\PAB\PABX-BVA\l-062-l\0621064.D Vial: 64 18 Oct 2000 10:20 Operator: MAN 2 x STD-5 Water Inst : MS7 MS Integration Parasis: RTEXNT.P Quant Method : D:\HPCHEM\l\METHODS\PABX_BVA.M (RTE Integrator) pbundanca 2000| i ! is 1000-: ! 50o! TIC: 0621064.0 l\ 1\ ii /' ;Tiim-> 6.10 6.2D_ 6jo 6.40 6.50 6.60 6.70 6.B0 6.90 7,00 7.10 730 7.30 740 7.50 7.60 770 7,80 7 90 MVL .. ........... r Ion': 131 (7.135) ........... .... Amount: ... .... ... 0.00___________________ 1I Reap: 2410 A }\ !\ 6.20 6.30 6.40 650 6.60 6.70 ' 6.80 690 7.00 7.10 rip 7.30 7.40 7.90 7.60 7.70 7.60 7.90 d4 Amount : 0.00 Ion: 135 (7.158) 1Resp: 4094 A !\ IV r-n T 6 20 6-30 640 630 6.60 6.70 6.B0 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7. 7.70 7.00 7 0 6 2 1 0 6 4 . D PABX BVA.M Wed O c t 18 1 3 : 0 0 : 2 6 2000 Page 2 001405 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-182 Page 181 Peak Area Report Sam ple Mama: 2 x STD-6 Water Notebook Reference: PABX-BVA-1-062 D ale A cquired: 10/18/2000 10:40 O p erator MAN Data File Nam e: 0621065.D Dale File Path: D\M SD_DATA\PAB\PABX-8VAU-062-1\ Inatram ent Method Name: PABX_BVA Inalm m ent Nam e: MS7 Inatram ent V M N um ber 65 Comwinrt f 1) 2) CanPO Tm tw m t MVl MVL-44 Retention Time IM In.) 7.13 7.16 ten 131 135 PwhAns 5207 5437 Peak Area Ratio 957.7E-3 M* The latter V in thia column mdicataa that manual imapralion ama partormad on lhat compound. Print Date and Time: 10/18/200013:00 Page 1 001406 Primedica-Worcester Project number: PABX-BVA 418-018:PA GE 1-183 Page 182 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Mise D:\MSD_DATA\FAB\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 (RTB Integrator) fid ane i 3000j iI i! ! 2000 1000 TIC 0621065.D t\ I M !I /l 610 6.20 830 640 8.80 660 6.70 8BO 690 7.00 7.10 7.20 70 7.40 7.SO 7.60 770 7B0 730 MVL _________________ Amount: .0 ___ !.Ion': 131 (7.134) j Reap: 5207 ''T- ' ! 6.20 6.30 6.40 6.50 660 6.70 6.80 d4 m .Ion: 135 (7.161) ;Resp: 5437 650 7.00 7,10 7Jg) 7,30 7.40 7SO Amount : 0.00 -- I 1I 1 7,60 7.70 7.60 7.90 IS !\ 620 6.30 6.40 650 6.60 6.70 6.B0 6.90 7.00 710 7.20 7.30 7.40 750 7.60 7.70 7J0 790 0 6 2 1 0 6 5 . E> PABX BVA .M Wed O c t 18 1 3 : 0 0 : 4 5 2 0 0 0 Page 2 001407 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-184 Page 183 Peak Area Report Sample Name: 2 STD-7 Water Notabook R ateane : PABX-BVA-1-062 Data Acquired: i o n 8000010:59 O perator MAN Data FUe Name: 0621066.0 Data File Path: D:\MSO_DATA\PAB\PABX-BVA\1-062-1l Instrum ent Method Name: PABX EVA hutnim ent Name: MS7 hutnim ent Vial Number: 66 Compound # 1) 2) Compound Name MVL MVL-4 Retention Tim e (M in.l 7.14 7.16 jOQ 131 135 Peak Area j 11012 8581 Peak Area Ratio 167.8E-2 M* * The letter In " in this coturni Indicalea that manuel attestation area portem eli on that compound. P rint Date and Time: 10/18/2000 13:00 Page 1 001408 Primedica-Worcester Project number PABX-BVA 418-018:PAGE 1-185 Page 184 Quantitation Report (QT Reviewed) Data Pile Acq On Sample Miec 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:\HPCHBM\1\METH0DS\PABX_BVA.M (RTB Integrator) 0 6 2 1 0 6 6 . D PABX BVA.M Wed O c t 18 1 3 :0 1 :0 4 2000 Page 2 001409 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-352 Page 166 Peak Area Report Sam ple Name: 11037-GROUP11-F1 Notebook Reteranca: PABX-BBA-1-047 Date Acquired: 1/3/01 14:49 O perator Data File Name: 0471082.D Data FHe Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Inatrum ent Method Name: PABX_BBA tnatnim ent Name: MS7 Inatnim ent Vial N um ber 82 Comoound 1) 2) Comoound Name MVL MVL-04 Retention Tim e (M in.l 7.02 7.04 OQ 131 136 Peak A rm M* 187 1562 Peak Area Radc. 119.7E-3 M*>The tener *nT In this column Indicatee that memiel Imagiallowwee pertormed en the compound. Pnnt Date and Time: VI8/01 15:11 Page. 001410 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-353 Page 167 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\1 -047-1\04710B2 .D Vial: 82 3 Jan 2001 14:49 Operator: 11037-GROUP11-F1 Inst : MS7 kS Integration Params: RTEINT.P Ruant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) V L __________________ __________________ Amount : ______ 0_.00 " ion: 131 "(7. OlsT Resp: 187 0-.71082.D PABX BBA.M Thu Jan IS ?! -.2:07 2: ' Page :> 001411 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-354 Page 168 Peak Area Report Sample Name: 2 X OC HIGH Notebook Reference: PA8X-BBA-1-047 Date Acquired: 1/3701 15:09 O perator Data File Name: 0471011.0 Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ Instrum ent Method Name: PABX.BBA tnatnim ant Name: MS7 Instrum ent Vial N um ber 11 Comoound * U 2) Comoound Name MVL MVL-d4 Retention Tim e (M in.) 7.02 7.04 las 131 135 Peak Area M* 1874 1498 Peak Area Ratio 125.1E-2 M* The lattar *m* m this column indiestse that manual mgialien was psttonnsO on that compound. Print Dale and Time: 1/18/01 14:48 Page 1 001412 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-355 Page 169 Daca File Acq On Sample Mise yuancitacion nepuiL ivi r c v c c u / D:\MSD DATA\PAB\PABX-BBA\1-047-1\0471011.D Vial: 11 3 Jan 2001 15:09 Operator: 2 X 0C HIGH Inst : MS7 0477011.D PABX BBA.M Thu J-rn 18 14:^3:09 2001 2 00141-3 Primed'.ca-Worcester Project Number: PABX-BBA 418-018:PAGE 1-356 Page 170 Peak Area Report Sample Name: 2 X BLANK Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3401 1529 O p erato r. Data File Name: 0471016.D Data FHe Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ Instrument Method Name: PABXJ3BA Instrum ent Name: MS7 Inatrum ent V ial N um ber 16 Comoound i) 2) Comoound Name MVL MVL-4 Retention Tim e (M in.) 0.00 0.00 Ion 131 135 Peek Area If* 0 0 Peak Area Ratio DIVA M - The letter *m* In th is colum n Ind icates that m snual k its y s tip n w as perform ed on that ecenpound. h t Data anc* : ime: 1/18/C. 4:49 Page 1 001414 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-357 P ag e 171 Vuantitacion neport igr aevieweaj Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471016.D Vial: 16 3 Jan 2001 15:29 Operator: 2 X BLANK Inst : MS7 tS Integration Params: RTEINT.P luant Method : D:\HPCHEM\l\METHODS\PABX_BBA.M (RTE Integrator) MVL Amount : Not Pound 0471016.1) PABX EiA.M fhu Jan )0 14:49:48 7001 Page 001415 Pnmedica-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 Acquired: 1/3/01 15:42 O perator Data File Name: 0471017.D Data File Path: D:\MS0_DATA\PAB\PABX-BBA\1-047-1\ kistrum ent Method Name: PABX_B8A Instrument Name: MS7 Instrum ent Vial Num ber 17 Comoound # 1) 2) Comoound Name MVL MVL-44 Retention Tim e (M in.) 0.00 7.04 jpn 131 135 PeakArea IT 0 1533 Peak Area Bado 000.0E-3 M* * The letter "m* In PUBcolumn Indicates that manuel Integration ama performed on that compound. Print Dat: :id Time: 1/1E/01 14:49 2 Page 1 001416 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-359 Page 173 Data File Acq On Sample M lB C Uuantitatxon Keporc tur Kevieweaj D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471017.D Vial: 17 3 Jan 2001 1S:49 Operator: 2 X CONTROL Inst : MS7 Amount: Not Found 0471017.D PABX_BBA.M Thu Jan 18 14:5' 07 2001 Pr 2 001417 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 Acquired: 1/3/01 16:09 O p erator Data File Name: 0471018.D Data R ie Path: D:\MSD_DATA\PAB\PA0X-BBA\1-O47-1\ Inatrum ent Method Name: PABX_BBA toatnim ent Nam e: MS7 m etrument Vial N um ber 18 Comoound # O 2) Comoound Name MVL MVL-4 Retention Tim * (M in.) 7.02 7.04 !aa 131 135 Peak Ama M* 138 1688 P *ak Area Ratio B17.5E-4 M* Tha iettar "m* in tMa column indica-- that manual imagil i on mo performed on that compound. I Print Date and Time: 1/10/01 14:50 J' Page 1 001418 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-361 Page 175 Quantitation Report tor Reviewed) Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\047101B.D Vial: 18 3 Jan 2001 16:09 Operator: 2 I STD 1 Inst : MS7 Integration Params: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) MVL_ ___ ____ _________________ Amount: : Ion :*"3"*T7.017)" !Reap: 138 _0.00 0471018.D PABX BBA.M Thu Jan 18 14: SO -.17 2001 ige 2 001419 Primedica-Worcester Project Number: PABX-BBA 418-018-.PAGE 1-362 Page 176 Peak Area Report S im ple Name: 2 X STD 2 Notebook R ite n n e : PABX-6BA-1-047 D ite Acquired: 1/3/01 1629 O p erator: Date File Name: 0471019.D Date File Path: D:\MSD_0ATA\PAB\PABX-BBA\1-047-1\ Inatnim em Method Name: PABX_BBA Inetnim ent Name: MS7 bistnim ent Vial N um ber 19 ComDOund # 1) 2) Com ootm d Name MVL MVL-d4 Retention Tim e (M in.) 7.02 7.04 ton 13t 135 Peak A n a M* 229 1538 Peak Ama Ratio 14B2E-3 M* The letter *nT In Mite coturno IndicateOiat manual Integn tten ne pattennad an K it compound. Phnl Date i i.u Time: 1/18/01 14:50 Page 001420 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-363 Page 177 Quantitation Report (QT Revieveat Data File Acq On Sample Mise D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471019.D Vial: 19 3 Jan 2001 16:29 Operator: 2 X STD 2 Inst : MS7 Integration Parame: RTEINT.P ant Method : D:\HPCHEM\1\METH0DS\PABX_BBA.M (RTE Integrator) MVL____ _ ____ _____ _ Amount _____ 0 .00 Ion: 13 'ii~ .'0 2 ) ........................ .... . 0471019.D PABX BBA.M Thu Jan 18 14:50:47 2001 Page 2 001421 Prixnedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-364 Page 1/8 Peak Area Report Sample Name: 2 X STD 3 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 16:49 O perator Data File Name: 0471020.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1-047-1\ kiatrum ent Method Name: PABX_BBA kiatrum ent Name: MS7 kiatrum ent Vial Number. 20 Comoound 1) 2) Comoound Name MVL MVL-4 Retention Time (M in.) 7.02 7.04 ton 131 135 Peak Area M ' 386 1311 Peak Area Ratio 294.4E-3 M' The letter'm* In ml* column indieelae Piet menual InlopiotKin rae performed en Piet compound. Print Date and Time: 1/18/01 14:50 Page 1 001422 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-365 Page 179 Data File Acq On Sample Mise yuantitation Report iui Reviewed) D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471020.D Vial: 20 3 Jan 2001 16:49 Operator: 2 X STD 3 Inst : MS7 C471020.D PABX BBA.M Thu Oan 18 14:51:06 2001 Page 2 001423 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-366 Page 180 Peak Area Report Sample Nam e: 2 X STD 4 Notebook Reference: PABX-BBA-1-047 Date A cq uim i: 1 /3 D 1 17:09 Operator: Data R ie am a: 0471021.D Data FUa Path: D:\MSD_DATA\PAB\PABX-BBAV1-047-1\ instrument Method Name: PABX_BBA Instrum ent ama: MS7 tnatnim ant Vial N um ber 21 Comeound * 1) 2) Com oound Name MVL MVL-d4 R etention Tim e IM In.t 7.02 7.04 lea 131 135 P ik Arca M* 565 1651 Peak Area Rado 342 -3 M* The letter *m* In this column Indlcatts that manual Integration es psrtormsO en atei compound. Print L iiie and Time: 1/M>/01 14:51 Page ?. 001424 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-367 P ag e 181 Data File Acq On Sample Mise Quantitation Keport iwi itevieweaj D:\MSD DATA\PAB\PABX-BBA\1-047-i\0471021.D Vial: 21 3 Jan 2001 17:09 Operator: 2 X STD 4 Inst : MS7 0471021.D PABX_BBA.M Thu Jar 18 14:51:26 2001 Prre 2 001425 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-368 Page 182 Peak Area Report Sample Name: 2 X STD 5 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 1729 O p erator. Data File Name: 0471022.D Data File Path: O:\MSD_DATA\PAB1PABX-BBA\1-047-1\ Instrum ent Method Name: PABX_BBA kwtrum ent Name: MS7 Inatnim ent Vial Number. 22 Comoound # t) 2) Com oound Name MVL MVL-d4 Retention Time fM ln.l 7.02 7.04 Ion 131 135 Peak Area M* 936 1488 Peak Area Ratio S29.0E-3 M * mThe letter ama In this column Indicate* that manual Integration am * performed on etat compound. Print Date and Time: 1/18/01 14:51 Page 1 001426 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-369 Page 183 Quantitation Report (QT Reviewed) Data File Acq On Sample Mise 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 Integration Params: RTEINT.P ,t Method : D:\HPCHKM\1\METH0DS\PABX BBA.M (RTE Integrator) M V L _______ _______ 7 io n : 31 ~ff709)' Rasp: 936 __________ Amount : 0.00 0471022. D PABX BBA.M Thu Jan 18 14:51:47 2001 Page 2 001427 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-370 Page 184 Peak Area Report Sample Nam: 2 X STD 6 Notebook Reference: PABX-BBA-1-047 Date Acquired: 1/3/01 17:48 O p erator: Data R ie Name: 0471023.0 Data R ie Path: D:\MS0_DATA\PAB\PABX-B8A\1-047-1\ Instrument Method Name: PABX_BBA Inatrum ent Name: MS7 Instrument Vial Number: 23 C om pound 1) 2) Compound Name MVt MVL-04 Retention T im * fMin.1 7.02 7.04 Is a 131 135 Peak Area M* 1415 1389 Peak Area Ratio 101.9E-2 U * * The letter tn * In this celiim n jndlrefee Piet manuel Integration tree performed on feel compound. Print Date and Time: 1/18/01 14:52 Page 1 001428 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-371 Page 185 Data File Acq On Sample Mise Quantitation Report IT Reviewed) D:\MSD DATA\PAB\PABX-BBA\l-047-l\0471023.D Vial: 23 3 Jan 2001 17:48 Operator: 2 X STD 6 Inst : MS7 M V L ____ _______ ____________________ Amount :________ 0.00 Ion: 13 (7.0207" 0471023.D ?ABX BBA.M Thu Jan 18 '4:53:03 2001 Page 2 00149 Primedica-Worcester Project Number: PABX-BBA 418-018:PA GE 1-372 Page 186 Peak Area Report Sample Name: 2 X STD 7 Notebook Retarenee: PABX-BBA-1-047 Data Acquired: 1/3XJ1 18:08 O perator Data File Name: 0471024.D Data File Path: D:\MSD_DATA\PAB\PABX-BBA\1 -047-1\ kiatnim ent Method Name: PABX.BBA tnetnim ent Name: MS7 biabum ent Vial N um ber 24 Comoound # 1) 2) Com oound Name MVL MVL-d4 Retention Tlm etM in.l 7.02 7.04 ba 131 135 PeakAraa I f 2396 1444 Peak Ama Rado 16S.9E-2 M- mThe letter V in triti column mate rn e Piet manual mt e y nen ewe pertermea en diet compound. Print Date and Time: 1/ 18/01 14:53 Page 1 001430 Primedica-Worcester Project Number PABX-BBA 418-018:PA GE 1-373 Page 187 Data File Acq On Sample Mise Uuantitation Report ICT Reviewed) D:\MSD DATA\ PAB\PABX-BHA\l-047-l\0471024 .D Vial: 24 3 Jan 2001 1B:08 Operator: 2 X STD 7 Inst : MS7 #v MVL__ _ ________________________________ Amotine :______ 0.0 : ioti: ill '(7.017) : Resp: 2396 PABX BB.K Thu Jan 16 14:53:23 2001 Page 2 001431 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 Fieport: FACT TOX-169 LRN-E01-0180 Study Title Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) in the Mevalonic Acid/Cholesteroi 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 J u ly 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 3 M Environm ental Laboratory . 'A / Environmental Laboraton- 001432 Page 1 Page 1 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 4I8-018:PA G E 1-375 Analytical Report: FACT-TOX-169 LRN-EO1-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 This page has been reserved for specific country requirements. 3M Environmental Laboratory S \l Environmental Laboraron 001433 Page 2 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-EO1-0180 Analytical Report: FA C T TO X -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, FACTTOX-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., S tu d y D ire c to r (p/jfiln I Date ^ T iriv i*-L T * iMarvin T. Case, D.V.M., Ph.D., S p o n s o r R e p r e s e n ta tiv e l Date Kristen J. Hansen, Ph.D., P rin c ip a l A n a ly tic a l In v e s tig a to r Jl,vrX. Date T'dV I William K. Reagen, Ph.D., L a b o r a to r y M a n a g e r TMs-/os 001434 Cate 3M Environmental Laboratory 5M Environmental Laboratory Page 3 Page 3 418-018:PA GE 1-377 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 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-E01-0180 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 Q A U R ep resen tative 6 - 11-CM Date 3M Environm ental Laboratory -'\! Environmental Laboratori' 001435 Page 4 Page 4 418-018:PA G E 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-E01-0180 Analytical Report: FACT TO X -169 LRN-E01-0180 GLP Compliance Statement....................................................................................................... 3 GLP Study-- Quality Assurance Statement..............................................................................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.......................................................................3Q 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)........................46 Appendix D: Data Summary Tables............................................................................................57 Appendix E: Data Spreadsheets................................................................................................. 53 Appendix F: Example Calculations............................................................................................. 63 Appendix G: Interim Certificate of Analysis............................................................................. 64 Appendix H: Report Signature Page....................... 68 3M Environmental Laboratory }M Environmental Laboratory 001436 Page 5 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 418-018)...............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 TO X-169.............................................................................................................. 16 Table 6 . Characterization of the Test Article in Study FACT TOX-16 9 ............................... 16 Table 7. TOX 169 Data Summary of PFOS Concentration-- Rat Serum (pg/mL)............. 57 3 M Environm ental Laboratory' SM Environmental Laboratory 001437 1 age 6 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 D irector Deanna J. Luebker, M.S. 3M Corporate Toxicology (Medical Department) Building 220-2E-02 St. Paul, MN, 55144 Analytical C hem istry Laboratories Sera Analyses 3M Environmental Laboratory (3M Lab) Kristen J. Hansen, Ph.D., Principal 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 R epresen tative S era Extractions Pace Analytical Services, 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 Laboratori- 001438 ! age 7 Page 7 418-018:PAGE 1-381 3M Medical Department Study: T-6295.25 3M Medical Department Study; T-6295.25 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Analytical Report: FACT TOX-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. Test 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). T able 1. Fem ale R at F0 Population Dem ographics fo r Study (Argus 418-018) Dosage Groups P o p u la tio n Selected fo r S tu d y Dosage V ehicle Control G roup 1 M evalonic Acid Control G roup 2 Group 3 Group 4 Cholesterol Control G roup 5 Group 6 Group 7 G roups G roup 9 Group 10 G roup 11 G roup 12 Group 13 28 28 28 28 28 28 28 20 20 20 20 28 28 0 .5 % Tw een 80 m evalonic acid 1.6 m g/kg P F O S + m evalonic acid 2 m g/kg P F O S -t- m evalonic a d d cholesterol 1.6 m g/kg P F O S + cholesterol 2 m g/kg P F O S + cholesterol 0 .4 m g/kg P FO S 0 .8 m g/kg P FO S 1 m g/kg P FO S 1.2 m g/kg P FO S 1.6 m g/kg P FO S 2 mg/kci P FO S 001439 3 M Environm ental Laboratory 3 \l Environmental Laboratory- Page 8 Page 8 418-018:PAGE 1-382 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Analytical Report: F A C TTO X-169 LRN-E01-0180 Sp ecim en C o lle c tio n and A n a ly sis 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 Specimens-- 54 specimens (FO Females, GD21, Caesarean section) Sera Specimens-- 54 specimens (F1 pooled litter, GD21, Caesarean section) Sera Specimens--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-ESM8MS) 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 Environmental Laboratory M Environmental Laboratory 001440 ' age 9 Page 9 3M Medical Department Study: T-6295.25 3MMedical Department Study: T-6295.25 418-018:PAGE 1-383 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 Chemical Characterization of the Reference Standard P o ta s s iu m P e rflu o ro o c ta n e s u lfo n a te (KPFO S) CAS Number 2795-39-3 Chemical Formula: CsF,7S0 3'IC Molecular Weight: 537.9 Chemical characterization information on the reference substance potassium perfluorooctanesulfonate used in this study is presented in tabular forni below. T ab le 2 . C haracterization o f th e Analytical R eference Substance In S tudy FACT TO X -169 Reference Substances Chemical Nam* Source Expiration Data Storage Conditions Chemical Lot Number Physical Description KPFOS (Potassium Perfluorooctanesulfonate)* 3M CorporateToxicology, Medical Department 8/31/01 Frozen S-10C 217 White crystalline powder THPFOS (Tetrahydrooerfluorooctanesulfonic add) TCR00017-055 SynQuest Labs 2005 -20C 075-91 White powder Purity 86.9% TBD* T erge! analyte Is PFOS, CtF,TSOj` TBD--To be determined 3M Environmental Laboratory .'M E n viron m en tal L aboraron - 001441 Page 1U age 10 418-018:PA GE 1-384 3M Medical Department Study: T-6295.25 3MMedical 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 P r epa r a t o r y M eth o d 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. A n a l y t ic a l M e t h o d 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 (CaFnSOa-) analysis, was fragmented to produce ion 99 (FSO3-). The characteristic ion 99 was monitored for quantitative analysis. 3M Environmental Laboratory M E nvironm ental L aboratory 001442 Page 11 P a g e 11 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 418-0I8:PA G E 1-385 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 A n a l y t ic a l E q u ip m e n t The following is representative of the settings used during the analytical phase of this study. Liquid Chromatograph: Hewlett-Packard* Series 1100 Liquid Chromatograph system Analytical column: Keystone* BetasilTM Cu 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 (m in u te s ) 0.0 1.0 5.5 7.5 8.0 %B 10% 10% 95% 95% 10% Mass Spectrometer: Micromass* API/Mass Spectrometer Quattro IITMTriple Quadrupole system Software: Mass LynxTM 3.4 Cone Voltage: 60 V Collision Gas Energy: 40-45 eV Mode: Electrospray Negative Source Block Temperature: 150C 10C Electrode: Z-spray Analysis Type: Multiple Reaction Monitoring (MRM) Table 3. Target Ions Monitored in 3M Laboratory Analyses Target Analyte Prim ary Ion (a m u ) Product Ion (a m u ) PFOS 499.0 99.0 THPFOS 427.0 80.0 3M Environmental Laboratory SM E nvironm ental L aboratory 001443 Page 1. age 12 418-018:PAGE 1-386 3M Medical Department Study: T-6295.25 JM Medical Department Study: T-6295.25 A nalytical R eport: FA C T -T O X -169 L R N -E O 1-0180 Analytical Repart: FACTTOX-169 LRN-E01-0180 D eviations 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 (ra) 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 M Em ronmental Laboraron.- 001444 Page 13 P age 13 418-018:PAGE 1-387 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 A nalytical Report: FACT-TOX-169 L R N -E 01-0180 Analytical Report: FACTTOX-169 LRN-E01-0180 Sum m ary o f Q u ality C ontrol A n a ly se s R esults L in e a rity : T h e coefficient of determ ination (r2) of the standard curve w a s 0 .9 9 0 . C alibration S tan dards: Quantitation of the target analytes w as based on linear regression analysis (1/x weighted) of an extracted rat sera matrix curve run prior to each group of sam ples. High or low points on the curve m ay 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 m ay have been significantly affected by background levels of the analyte. Occasionally, a single m id-range curve point that was an obvious outlier m ay have been deactivated. Q uantitation of each analyte w as based on th e response of one specific product ion using the multiple response-monitoring m ode of the instrument (see Appendix C, Analytical M etho d s). L im its o f Q u a n tita tio n (L O Q ): T h e L O Q is equal to the low est accep tab le standard in the calibration curve (defined as the lowest valid standard within 3 0 % of the theoretical value), an d is a t le a s t tw o tim es th e analyte p eak a re a detected in th e m atrix blanks. (S e e below.) T ab le 4 . D eterm inations o f th e LO Q in the Analyses o f P FO S Extracts Analyte Method LOQ PFOS 0.00494 pg/mL B la n k s : All blanks w ere below the low er limit of quantitation fo r the com pounds of interest. P recision: Precision w as determined by analysis of extracted calibration check standards, which w ere reproducible to within 2 5 % over the valid linear range. in s tru m e n t P recisio n: Instrumental precision was determ ined by replicate (n=7) injections of a single serum extract. P eak area was reproducible to within 2% . In te rn a l S ta n d a rd s : T h e internal standard (T H P F O S ) w as added to all sam ples and standards. T H P F O S was not used for quantitation, but w as used to monitor for gross instrument failure. T he surrogate response of each analytical run w as verified to determine that it did not vary more than 5 0 % from the m ean within each analytical run. S am ples with deviant internal standard response w ere confirmed by reanalysis and are m arked in the table of results. Statem ent o f D ata Q uality It is not possible to verify true recovery of endogenous analyte from tissues without radio-labeled reference material. The only m easurem ent of accuracy available at this tim e, extracted calibration curves and multi-level continuing calibration verifications (C C Vs), indicate that the sera data are quantitative to 25% or greater. 3M Environmental Laboratory SM Environm ental L aboratori 001445 Page 14 P age 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: FACTTOX-169 LRN-E01-0180 Sum m ary o f Sam ple R esu lts P FO S results have been corrected for purity of the analytical reference material. S a m p le s fro m C o n tro l A n im a ls : Low levels of P F O S w ere often d etected in th e sera of the control anim als. T h e s e levels w ere significantly low er than those fo und in th e low d ose test an im als. S a m p le s fro m D o s e d A n im als: In general, P F O S levels found in the s e ra o f th e test anim als increased with dose level. D etailed sam ple d ata tables are presented in A ppendices D and E. Statistical Methods and Calculations Statistical methods w ere limited to the calculation of m eans and standard deviations. S ee A ppendix F for exam p le calculations used to g enerate the serum sam ple d a ta in F A C T T O X 169. Statement of Conclusion U nd er the conditions of th e present studies, the fluorochemical P F O S w as observed in th e sera of all FO generation rats dosed with the test article and in all F1 generation rats from FO dosed rats. 3M Environmental Laboratory .'M E nvironm ental L aboratory 001446 Page 15 page 15 3M Medical Department Study: T-6295.25 3M Medicai Department Study: T-6295.25 418-018:PAGE 1-389 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Analytical Repart: FACTTOX-169 LRN-E01-0180 Appendix A: Control Matrices and Test Article Table 5. C haracterization o f the C ontrol M atrices Used fo r Sera Analyses in Study FACT TO X -169 Control Matrix fat Serum Rat Serum Rat Serum Rat Serum Source TCR# Expiration Date Storage Conditions Chemical Lot # Physical Description Sigma Chemicals 99062-9 01/01/2010 Frozen -20"C 17H9306 Rat Serum Sigma Chemicals 99062-31 01/01/2010 Frozen -20*C 17H9306 Rat Serum Sigma Chemicals 99131-9 01/01/2010 Frozen -20C 19H89291 Rat Serum Sigma Chemicals 99131-11 01/01/2010 Frozen -20C 19HB9291 Rat Serum T able 6. C haracterization o f th e T est Article in Study FACT TO X-169 Location 3M Laboratory Test Article Source Expiration Date Storage Conditions Chemical Lot Number Physical Description Purity Potassium Perfluorooctanesulfonate 3M Corporate Toxicology, Medical Department 8/31/01 Frozen s-10C 217 White crystalline powder 86.9% 3M Environmental Laboratory 3M E nvironm ental L aboratory 001447 Page 16 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 Environmental Laboratory 7M Environmental Laboratorv 001448 Page 17 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 P h a s e Title Determination o f the Presence and Concentration o f Perfluorooctanesulfonate (PFOS) in the Mevalonic Acid/Cholesterol Challenge and NOEL Investigation Study in Rats PHASE PROTOCOL Performing 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 Environmental Laboratory E nvironm ental L aboratory' 001449 Page 1 of 9 P age 18 418-018:PA GE 1-392 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 L R N -E 01-0180 Protocol F A C T -TO X -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 Director Phase Locations Principal Analytical Investigator (PAI) Analytical Testing Laboratories 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. 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN55106 Proposed Phase Timetable 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 Environmental Laboratory SM Environmental Laboratori- 001450 P ag e 2 of 9 Pp oargree 199 3M Medical Department Study: T-6295.25 418-018:PAGE 1-393 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol F A C T -T O X -169 1. Study Determination o f 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 o f PFOS in sera specimens of FO generation 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 CFRPart58. 4. Quality Assurance 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), CgFnSOjK, CAS 2795-39-3 6. Test System T h irte e n d o se g ro u p s o f fe m a le ra ts w e re g iv e n th e te s t article by oral (g a v a g e ) ro u te . W ith in each group, one blood sample was collected from six female FO on day 21 of gestation (at Caesarean sectioning), from six female FO on day 5 of lactation (after natural delivery), from six FI pooled litters on day 21 o f 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 o f all samples received. 3M Environmental Laboratory .'M E n vironm ental L a b o ra to n 001451 Page 3 of 9 Page 20 418-018:PAGE 1-394 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol F A C T -TO X -169 Number of Specim ens per Dosage group and Dosage Levels Dosage Group and Level 1 Vehicle Control 2 Mevalonic Acid Control 3 1.6 mg/kg PFOS * Mevalonic Acid 4 2.0 mg/kg PFOS Mevalonic Acid S Cholesterol Control 6 1.8 mg/kg PFOS Cholesterol 7 2.0 mg/kg PFOS Cholesterol 8 0.4 mg/kg PFOS 9 0.8 mg/kg PFOS 10 1.0 m g/kg PFOS 11 1.2 m g/kg PFOS 12 1.6 m g/kg PFOS 13 2.0 m g/kg PFOS F0 Rats from Caesarean Section Day 21 6 6 6 6 6 6 6 6 6 6 6 6 6 Ft Rat Pups from Caesarean Section Day 21 6 6 6 6 6 6 6 6 6 6 6 6 6 FO Rats from Lactation Day 5 6 6 6 6 6 6 6 6 6 6 6 6 6 F1 Rat Pups 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. Details of 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 o f methanol and filtered through a 3 cm3plastic syringe attached to a 0.2 pm nylon filter into glass autovials. 001452 3M Environmental Laboratory -\\1 E nvironm ental L aboratory' Page 4 of 9 P age 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 -TO X -169 9. Analytical Methods ETS-8-5, "Analysis of Potassium 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 (CgFnS03-) analysis, is fragmented further to produce ion 99 (FSO3-). 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. D a t a Q u a l i t y O b j e c t i v e s 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 Limits of Quantitation (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 Acceptable Precision Quality control samples are required to meet 25% precision, provided they are quantitated within the selected calibration range. 10.4 Use of Confirmatory Methods I f a confirm atory m ethod is used, an am endm ent to this protocol w ill be w ritten. 10.5 Demonstration of Specificity, 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 5 \I E nvironm ental L aboratori' 001453 Page 5 of 9 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 -1 69 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 ofN-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 andCalibration 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/Thermooouples PSS-MTR-06 Verification of Calibration of the EppendorfPipettor 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 of the 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. 001454 3M Environmental Laboratory 3M E nvironm ental L aboratory' Page 6 of 9 Page 23 418-018:PA GE 1-397 3M Medical Department Study T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protoco/ 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 ofthe 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 o f 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 OFRAWDATA, RECORDS, ANDFINAL REPORT Original data or copies thereof, will be available at 3M Environmental Laboratory to facilitate audits of the study during its progress and b efo re acceptance of the phase report. When the phase report is completed, all original paper data, including those items listed b e lo w w ill b e re ta in e d in th e a rc h iv e s o f 3 M E n v iro n m e n ta l L a b o ra to ry . A ll c o rre sp o n d in g 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. 1 4 .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: Vi Environm ental L aboratory SM E nvironm ental Laboratory' Page 7 of 9 Page r4 418-018:PA GE 1-398 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Protocol F A C T -TO X -169 14.2.1 14.2.2 14.2.3 14.2.4 Training records Calibration records Instrument maintenance logs Standard Operating Procedures, Equipment Procedures, and Methods 15. Data / S ample Retention All 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 S t Paul, MN 55144 Telephone (651) 733-5070 Fax (651) 737-4754 16. Protocol Amendments and deviations 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. 1 7 . A TTAC H M EN TS 17.1 Attachment A: Material Safety Data Sheets (MSDS) for Reference Standard '<M Environmental Laboratory -M Environm ental Laboratori' 001456 Pace 8 of 9 Page J7 3M Medical Department Study: T-6295.25 18. S ignatures 418-018:PAGE 1-399 Analytical Report: FACT-TOX-169 ProtocolFACT-Td$$9E()1' 180 T & * - ________________ __________ i f M arvin T. Case. D. V.M., Ph.D ., Sponsor Representative Date L D eanna J. L u ebkerlM .S ., Study Director jJ S lJ Q L Date K risten J. H ansen, Ph.D., Principal Analytical Investigator Ot l& llo \ Date .Vf E n v iro n m e n ta l L a b o rato ry SAI E n viro n m en ta l L aboratory- 001457 Page 9 of 9 Page 26 418-018:PAGE 1-400 3M Medical Department Study: T-6295.25 Record of Deviation Analytical Report: FACT-TOX-169 LRN-E01-0180 /. Identification Study / Project No. FACT-TOX-169, Lims # EOI-0180, Argus 418-018 Deviation Type D SOP 0 Method QEquipment Procedure (Check one) X Protocol 0 Other: Document Number FACT-TOX-169 Date(s) of occurrence 02/01/01 II. Description: Required Procedure/process: The protocol states that 312 specimens will be received at the Environmental Laboratory. Actual Procedure/process: 235 specimens were received at the Environmental Laboratory. III. Actions Taken: (such as amendment issued, SOP revision, etc.) This deviation was written. Recorded By oL a tL*, IV. Impact on Study / Project No adverse impact on the outcome of this study. Date i. 0 5 / ZH f o Authorized By (Study D irector /P r o je c t Lead) Date . S /^ / l Slud D.ilcjior : Dta/wOk Lutti;tr Re-pr^U.. Mwvin 3A t E nvironm ental L aboratory F orm E T S-4-8.0 31^0**/ 001458 Deviation No. (assigned by Study Director or Project Lead at the end of study or project) .\M E nvironm ental L a b o ra to ri Page 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. Identification Study / Project No. FACT-TOX-169, Liras#EOI-0180, Argus418-018 Deviation Type SOP I Method Equipment Procedure (Check one) Protocol Other Document Number ETS-8-4.2 Date(s) of occurrence 2/15/01-2/21/01 II. Description: 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 11.1.2 Do not extract less than O.SmL 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. Actions Taken: (such as am endm ent issued, SO P revision, etc.) This deviation was written. Recorded By Date i Kelly Swartout i d X u u x i a j p l i n J IV. Impact on Study / Project 102/21/01 a la a J o ) 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. 0 2 . I 2 Z l. I q i Authorized By (Study D irector /P r o je c t Lead) Date 3 M E nvironm ental L aboratory Deviation No. F orm E T S-4-8.0 (assigned by Study Director or Project Lead at the end of study or project) 001459 Dtrt^W . Kri* Ofc&i'ftfr. Lutiik^r 3M Environmental Laboratory -2/e?"7 fo I 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 I. Identification Study / Project No. Deviation Type (Check one) ' T r f / X t SOT TSfMethod Equipment Procedure Protocol CXOther. Document Number IS Date(s) of occurrence IL D escription: Required Procedure/process: --(-H-- t'YZ/sOonCC_____ M- S^- 7 p/tX'h -fiiij /I n d ie s S /?s~ s - / 9 s y z .- -i b / H j / lT j . w j j n /y, f i t s t * ------- -- ^ry z- r-i-c-^--L---siT 1/ ' f a r e J "rfi .Z f S S i' f o / A z J r / S - , [^ 2 A ^ y O J X - rt? Actual Procedure/process: l o O /j s ^ t s > XL J i / j A x J - ^ ^ /lyyy ^ / y / / >-3 / 1 __ /2 rL fa * 7 S sl - PS/A . / A I X * L i * / so C > r?/77'> ~ > ^OX/ /O sssstM * % *T III. A ctio n s Taken: __________ (such a s am endm ent issued, S O P revision, etc.) . /to n & j n . TV- /S h c i? a / //B / M /^ W i Recorded By N o adre***- IV. impadt on^tudy / Project QHlo^lQI Authorized By (Study D irecto r / P roject Lead) Date >^tnur Rifrticr'liiW.. fAtrv Ca., 3 M E nvironm ental Laboratory? )^ F orm E T S-4 -8 .0 / 'C L }.\1 Environmental Laboratory tucla /tctor '- Dlnno. lueoltr 4jk(*t\Ar&-*2&.4 jb & p W t O f T 4 _ 9t * ( Deviation No. _ ' (assigned by Study Director or Project Lead attththeeeei nd of study or project) 1 4 60 Page 29 418-018:PAGE 1-403 3M Medical Department Study: T-6295.25 3MMedical Department Study: T-6295.25 Analytical Report: FA.CT-TOX-169 LRN-E01-0180 Analytical Report: FACTTOX-169 LRN-E01-01B0 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 Ruorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry, (11 pages) SM Environm ental L aboratory 001461 Page 30 3M Medical Department Study: T-6295.25 418-018:PA GE 1-404 Analytical Report: FACT-TOX-169 LRN-E01-0180 3M Environmental Laboratory M ethod E xtraction o f F lu o ro ch em ica l compounds from Seru m fo r A nalysis Using H PL C -E lectrospray/M ass Spectrom etry Method Number: ETS-8-4.2 Adoption Date: 03/01/99 Effective Date: L/2. 1 Approved By: Laboratory Manager -H-- Group Leader 0 2 , 2 /C '/ Date Oi'IOi./O 1 Date 1.0 Scope and Application 1.1 Scope: This method is for the extraction of fluorochemical compounds from serum. 1.2 Applicable compounds: Fluorochemical surfactants or other fluorinated compounds. 1.3 F t_ mX0) u (7 a Matrices: Rabbit, rat, bovine, monkey, and human serum or other fluids as designated in the validation report. 0 2.0 Summary of Method This performance-based method describes the procedure for extracting perfluorooctanesulfonate (PFOS) or other fluorochemical surfactants from serum, or other fluids, using an ion pairing reagent and methyl-re/t-butyl ether (MtBE). An ion pairing reagent is added to the sample and the analyte-ion pair is partitioned into MtBE. The MtBE extract is removed and put onto a nitrogen evaporator until dry. Each extract is reconstituted in 1.0 mL of methanol, then filtered through a 0.2 pm nylon filter using a 3-mL plastic Word 6/95 3M Environm ental L aboratory ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 1 of 15 Page J i 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. (Application of this method to seven fluorochemicals was demonstrated: PFOS, PFOSA, PFOSAA, EtFOSE-OH, PFOSEA, M556, and a surrogate standard {see 3 . 0 D efin ition s}). These sample extracts are analyzed following method ETS-8-5.2 or other appropriate method. 3.0 Definitions________________________________________________________________ 3.1 PFOS: perfluorooctanesulfonate (anion of potassium salt) CgFl7S03' 3.2 PFOSA: perfluorooctane sulfonylamide CgFi7S02NH2 3.3 PFOSAA: perfluorooctane sulfonylamido (ethyl)acetate C8F|7S02N(CH2CH3)CH2C02* 3.4 EtFOSE-OH: 2(N-ethylperfluorooctane sulfonamido)-ethyl alcohol C8F17S 0 2N(CH2CH3)CH2CH20H 3.5 PFOSEA: perfluorooctane sulfonyl ethylamide CgFi7S02N(CH2CH3)H 3.6 M556: CgF,7S02N(H)(CH2COOH) 3.7 Surrogate standard THPFOS: 1H-1H-2H-2H perfluorooctane sulfonic acid 4.0 Warnings and Cautions____________________________________________________ 4.1 Health and safety warnings 4.1.1 Use 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 known at this time. 6.0 Equipment_______________________________________________________________ 6.1 The following equipment is used while performing this method. Equivalent equipment is acceptable. 6.1.1 Vortex mixer, VWR, Vortex Genie 2 6.1.2 Centrifuge, Mistral 1000 or IEC 6.1.3 Shaker, Eberbach or VWR 6.1.4 Nitrogen evaporator, Organomation 6.1.5 Balance ( 0.100 g) 7.0 Su and M aterials_________________________________________________ ,__ 7.1 Gloves 7.2 Eppendorf or disposable pipettes, plastic or glass (or equivalent) 7.3 Polypropylene bottles, capable of holding 250 mL and 1 L (Nalgene or equivalerli) 001463 3M E nvironm ental L aboratory ETS-8-4.2 Extraction of Fluorochemicals from Serum . age 2 of i Page J ' 3M Medical Department Study: T-6295.25 418-018:PAGE 1-406 Analytical Repon: FACT-TOX-169 LRN-E01-0180 7.4 Volumetric flasks, glass, type A 7.5 40 mL glass vials (I-CHEM or equivalent) 7.6 Centrifuge tubes, polypropylene, 15 mL 7.7 Labels 7.8 Bottle-top Dispenser - 3.0 to 10.0 mL or a 5-10 mL graduated pipette, glass 7.9 Syringes, capable of 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 Note: Prior to using glassware and bottles, rinse 3 times with methanol and 3 times with water. Rinse syringes a minimum of 9 times with methanol, 3 rinses from 3 separate vials. 8.0 Reagents and Standards_________________________________________________ 8.1 Reagent grade water, Milli-QTM, Nanopure II or equivalent 8.2 Sodium hydroxide (NaOH), J.T Baker or equivalent 83 Tetrabutylammonium hydrogen sulfate(TBA), Kodak or equivalent 8.4 Sodium carbonate (Na2C 03), J.T. Baker or equivalent 8.5 Sodium bicarbonate (NaHCCh), J.T. Baker or equivalent 8.6 Methyl-T-Butyl Ether, Omnisolv, glass distilled or HPLC grade 8.7 Methanol, Omnisolv, glass distilled or HPLC grade 8.8 Serum or blood, frozen from supplier 8.9 Fluorochemical standards 8.9.1 KPFOS (PFOS) (3M Specialty Chemical Division), molecular weight = 538 8.9.2 PFOSA (3M Specialty Chemical Division), molecular weight = 499 8.9.3 PFOSAA+H (PFOSAA) (3M Specialty Chemical Division), molecular weight = 585 8.9.4 EtFOSE-OH (3M Specialty Chemical Division), molecuhir weight = 570 8.9.5 PFOSEA (3M Specialty Chemical Division), molecular weight = 527 8.9.6 M556 (3M Specialty Chemical Division), molecular weight = 557 8.9.7 Surrogate standard: 4-H, perfluorooctane sulfonic acid (1-H,l-H, 2-H, 2-H CgFuSOH, [THPFOS]), molecular weight = 428 8.9.8 Other fluorochemicals, as appropriate 0 l4 6 4 SM E nvironm ental Laboratory' ' ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 3 of If Page 33 3M Medical Department Study: T-6295.25 418-018:PAGE 1-407 Analytical Report: FACT-TOX-169 LRN-E01-0180 8.10 Reagent preparation NOTE: When preparing different volumes than listed in reagent, standard, or surrogate preparation, adjust accordingly. 8.10.1 10 N sodium hydroxide (NaOH): Weigh approximately 200 g NaOH. Pour into a 1000 mL beaker containing 500 mL water, mix until all solids are dissolved. Store in a 1 L polypropylene (or equivalent) bottle. 8.10.2 1 N sodium hydroxide (NaOH): Dilute 10N NaOH 1:10. Measure 10 mL of 10 N NaOH solution into a 100 mL volumetric flask and bring to volume using water. Store in a 125 mL polypropylene (or equivalent) bottle. 8.10.3 0.5 M tetrabutylammonium hydrogen sulfate (TBA): Weigh approximately 169 g of TBA into a 1 L volumetric containing 500 mL water. Adjust to pH 10 using approximately 44 to 54 mL of 10 N NaOH. (Add the last 1.0 mL of NaOH slowly, as the pH changes abruptly.) Dilute to volume with water. Store in a 1 L polypropylene (or equivalent) bottle. 8.10.3.1 TBA requires a check prior to each use to ensure pH = 10. Adjust as needed using 1 N NaOH solution. 8.10.4 0.25 M sodium carbonate/sodium bicarbonate buffer (Na2C 03/NaHC03): Weigh approximately 26.5 g of sodium carbonate (Na2C 03) and 21.0 g of sodium bicarbonate (NaHC03) into a 1 L volumetric flask and bring to volume with water. Store in a 1 L polypropylene (or equivalent) bottle. 8.11 Standards preparation 8.11.1 Prepare PFOS standards for the standard curve. 8.11.2 Prepare other fluorochemical standards, as appropriate. Multicomponent fluorochemical standards are acceptable (for example, one working standard solution containing approximately 1.00 pg/mL of PFOS, PFOSA, PFOSAA, PFOSEA, M556 and EtFOSE-OH). 8.113 Weigh approximately 100 mg of PFOS into a 100 mL volumetric flask and record the actual weight. For standards with K* or other salts, multiply by a correction factor. For example, the molecular weight of KPFOS = 538, and the molecular weight of PFOS = 499. Calculate the correction factor by using the following equation: molecular wt, PFOS ( 499 ) = 0.9275 ( C o rre c tio n fa c to r ) molecular wt. KPFOS ( 538 ) 8.11.4 Bring to volume with methanol for a stock standard of approximately 1000 pg/mL. 8.11.5 Dilute the stock solution with methanol for a working standard 1 solution of approximately 50 pg/mL. 1000pg/m x5m = 50 pg/mL lOOmL 8.11.6 Dilute working standard 1 with methanol for a working standard 2 solution of approx. 5.0 pg/mL. 001465 3 \I E nvironm ental L aboratori' ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 4 of 15 Page 34 3M Medical Department Study: T-6295.25 418-018:PAGE 1-408 Analytical Report: FACT-TOX-169 LRN-E01-0180 50 g/ mLxlOmL = 5.0 pg/mL 100 mL 8.11.7 Dilute working standard 1 with methanol for a working standard 3 solution of approx. 0.50 pg/mL. 8.12 Surrogate THPFOS stock standard preparation 8.12.1 Weigh approximately 50-60 mg of surrogate standard l-H.l-H, 2-H, 2-H, CgFnSOjH (THPFOS) into a 50 mL volumetric flask ancl record the actual weight. 8.12.2 Bring to volume with methanol for a surrogate stock of approximately 10001200 pg/mL. 8.12.3 Prepare a surrogate working standard. Transfer approximately 1 mL of surrogate stock to a 10 mL volumetric flask and bring to volume with methanol for a working standard of 100 pg/mL. Record the actual volume transferred. 9.0 Sample Handling___________________________________________________________ 9.1 All samples are received frozen and must be kept frozen until the extraction is performed. 9.2 Allow samples to thaw at room temperature or in lukewarm water prior to extraction. 10.0 Quality Control_________________________________________________________ 10.1 Solvent Blanks, Method Blanks and Matrix Blanks 10.1.1 Solvent Blanks: An aliquot of 1.0 mL methanol is used as a solvent blank. 10.1.2 Method Blanks: Following this procedure, extract two 1.0 mL aliquots of water and use as method blanks. 10.1.3 Matrix Blanks: Matrix blanks are prepared from one of three sources: 1) a study control matrix from a study control animal received with each sample set; 2) a commercially obtained sample of the same species as the study animals; or 3) a surrogate matrix, also obtained commercially, but of a different species than the study animal (e.g. if rabbit is used to generate standard curves and CCVs for a monkey or rat sera study). The matrix to use depends on what matrix is used for the curve. 10.1.3.1 Study control matrix curve--If the study control matrix is used for the curve, prepare two (2) matrix blanks using the study control matrix. 10.1.3.2 Commercially obtained (same species) matrix curve--If the curve is prepared using commercially obtained matrix in the same species as the study animal, prepare two matrix blanks using this same commercially available matrix. 0 1 4 6 6 ,'M E nvironm ental Laboratory' ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 5 of 15 Page 3 V 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 ussd to prepare the initial curve (i.e., either the study control matrix, commercial matrix of same species, or surrogate matrix). 001467 :'M Environmental Laboratory ETS-8-4.2 Extraction of Fluorochcmicals from Serum Page 6 of 15 Page 6 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 1 mL 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 a;; 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 1 at 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 that falls within the calibration curve range of 2.5 ng/mL-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. s.\I E nvironm ental Laboratory- ETS-8-4.2 Extraction of Fluorochemicals from Serum 001468 Page 7 of 1.5 Page ^/ 3M Medical Department Study: T-6295.25 418-018:PAGE 1-411 Analytical Report: FACT-TOX-169 LRN-E01-0180 Table 1 Approximate spiking amounts for standards and spikes Using 1.0 mL of matrix Working standard (approx, cone.) pL Approx, final cone, of 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 0.500 ppm 50.0 ppm 15 0.750 ppm 50.0 ppm 20 1.00 ppm *ppm*^tgftnL 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-tenbutyl ether. 12.12 Cap each sample and put on the shaker at a setting of 300 rpm for 2.0 minutes. 01469 ' 1/ E n v ir o n m e n ta l L a b o r a to r i ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 8 of 15 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: ______ mL of std x concentration of std ( fig/mL)_____ mLof std + mL of surrogate std + initial matrix volume(mL) Final concentration ( fig / mL)of PFOS in matrix 14.0 M e t h o d Pe 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. 14J Refer to section 14 of ETS-8-5.2 for method performance criteria. 0011*70 SM E nvironm ental L aboratori ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 9 of 15 Page 39 3M Medical Department Study: T-6295.25 418-018.PA G E 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 Re c o r d s __________________________________________________________________ 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' horn 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 2 0 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 001471 3A/ E n v iro n m e n ta l L a b o r a to r i ETS-8-4.2 Extraction of Fluorochemicals from Serum Page 10 of 15 Page 40 418-018:PAGE 1-414 3M Medical Department Study: T-6295.25 Extraction Worksheet ETS-8-4.2 Analytical Report: FACT-TOX-169 LRN-E01-0180 Studv # M atrix Box# W k/Dav Surrogate Std actual pg/m L # FC-M ix approx. 0.5 Pg/m L # FC-M ix approx. 5 Pg/m L # FC-M ix approx. 50 pg/m L # Com m ents V erified by: D ate Spiked/ A n aly st: MS MSD ---- - -- - -- - .- - -- - --- - ---- - -- - -- - - -- Blank ID # . ... . -- amfflIHL=______ ____ m L _ .... ................. L . Serum E xtraction M e t h o d ---------------------------------------- 1--------------------------------------------D ateA Initials----------1-----------------Vortex 15 k c ,_________________________________________________________________________________________ 1------------------ Pinette Matrix, strike with appropriate surrogatt-OiiFCMiX____________________M stm E____ m L -- 1------------------ Pioette I mL of 0.5 M TBA. pH 10. pH = Std. # 1 Pipette 2 mL of 0.25 NajCCV0.25M NaHCOs buffer Std. # 1 Dispense/pioette 5 mL of methvl-t-butvl ether TN-A- Dispenser ID #: Shake 20 mm. Shaker ID #: Shaker speed: Ccntrifuee 20-25 min. Cenlrifuee IQfc___________________________ CcmrifltfC Remove a 4 mL aliouot of oream cJavei----------------------------------------------------------------------- Put on Ni open F vanorator to dtvness Add methanol Volume mL TN-A- Vortex 30 sec. 1. Corn. Cal. Verifications used same matrix as for std curve. 001472 Appendix A: Extraction Worksheet .'.\I E n viron m en tal L a b o ra to ry ETS-8-4.2 Page 1! of 15 P age 41 3M Medical Department Study: T-6295.25 418-018:PAGE 1-415 Analytical Report: FACT-TOX-169 LRN-E01-0180 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 1 6.03 19.2 5 ng/mI^1000 ng/mL PFOSEA 5.71 18.2 5 ng/mL-1000 ng/mL M DL/LOQ values in rat, bovine, monkey, and human serum, and monkey plasm a were not statistically determined. Tw o curves in each o f 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 M D L and LOQ will be the same values as determined in rabbit serum. Please see LO Q Summary and M D L study in ETS-8-4.0 & 5.0-V -l for further information. Appendix B: MDL/LOQ Values and Summary ETS-& 4.2 Extraction of Fluorochcmicals from Serum .'\t Environmental Laboraton- 001473 Pag- 12 of 15 Page 42 3M Medical Department Study: T-6295.25 418-018:PA GE 1-416 Analytical Report: FACT-TOX-169 LRN-E01-0180 Compound: P ros Prepared range Rabbit Serum of standards (ng/mL) Full Range 0.995 - 978 Low Curve 4.94 - 248 High curve 97.8 - 978 1/X 0.995 - 978 LCR from curve (ng/mL) 24.8 - 978 4.94 - 248 97.8-978 4.94 - 978 % Recovery Range 83-108 85-104 85-106 94-111 Compound: PFOSA Prepared range Rabbit Serum of standards (ng/mL) Full Range 0.993 - 976 Low Curve 4.93 - 97.6 High curve 24.8 - 976 1/X 0.993 - 976 LCR from curve (ng/mL) 4.93 - 976 4.93 - 97.6 24.8 - 978 4.93 - 976 % Recovery Range 88-103 87-105 93-102 94-103 Compound: PFOSAA Prepared range Rabbit Serum of 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) 24.7 - 974 9.74 - 247 97.4-974 9.74 - 974 % 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 ancTSummary ETS-8-4.2 Extraction of Fluorochemicals from Serum S'M Environmental Laboratory 001474 'age 13 ol 15 Page 43 3M Medical Department Study: T-6295.25 418-018:PAGE 1-417 A nalytical Report: FA CT-TO X -169 I RN-E01-0180 Compound; EtFOSE-OH Prepared range Rabbit Serum of standards (ng/mL) Full Range 0.993 - 976 Low Curve 4 .9 3 -9 7 .6 High curve 49.3 - 976 1/X 0.993-493 C om pound: PI "OSEA 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 C om pound: h' [556 Prepared range Rabbit Serum of standards (ng/mL) Full Range 0.993 - 976 Low Curve High curve 1/X 4.93 - 97.6 97.6 - 976 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 86-111 % 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 11.1-21.2 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 :."rum SM Environmental Laboratory 001475 Pag' ! 4 of 15 Page 44 3M Medical Department Study: T-6295.25 418-018:PAGE 1-418 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Example Ion Pair Standard Curves - Fluids A n aly ses) Study num ber: V Prep date(s): A n aly te(s): Final solvent and TN : Blank fluid/identifier: ! R eu irj matrix used u*prq-are turves, ra g .. 'R abbit,' ID XXX) | Study Animal M atrix : M ethod/revision: T arget analyte(s): FC mix std approx. 0.500 ppm: FC mix std approx. 5.00 ppm: FC mix std approx. 50.0 ppm: fIDRecord, d.ie stand,ard,s used. to prepare tlie extracted. curves. .) 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 Ug/mL ug/mL ug/mL ug/mL ug/mL ug/mL Calculated concentrations of standards in the sera matrix: Rabbit PFOS PFOSA PFOSAA EtFOSE PFOSEA M556 Surrogate Final cone ne/mL 4.93 Final cone ne/mL SM Final cone ne/mL 5.24 Final cone ne/mL 4.94 Final cone ne/mL 5.01 Final cone ne/mL 5.13 Std cone ne/mL 100 9.76 9.89 10.4 9.78 9.93 10.2 24.8 -'1 49.3 13 5 0 .0 -- h 26.3 24.8 25.2 25.8 Surrogate. A t H Final cone 97.6 Z 248 !1 251 / 493 I # I 699 j 1 2 5 1 GL258> T^- ne/mL 500 735 1 746 782 737 ^ J 4 9 766 976 ! 989 1038 978 993 1017 AU Ain't spiked mL 0.005 V alidated ran g es --a p p ro x im a te concentratio n s Serum PFOS PFOSA PFOSAA Rabbit 5.00-1000 ne/mL 5.00-1000 ne/mL 5.00-1000 ne/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. E tF O S E -O H 5.00-1000 ne/mL PFOSEA 5.00-1000 ne/mL M556 5.00-1000 ne/mL i)01476 Attachment C: Ion Pair Standard Curves ETS-8-4.2 Extraction of Fluorochemicals from Serum ,'M Environmental Laboratory Page 15 of 15 Page 45 3M Medical Department Study: T-6295.25 418-018:PAGE 1-419 A nalytical Report: FA C T -T O X -169 L R N -E 01-0180 3M Environmental Laboratory V Method Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/M ass Spectrometry M eth od N u m b er: E T S -8-5.2 A uthor. L isa C lem en, Kris H ansen Approved By: _____ . / r t ---------------------------- ------------------------------------- Laboratory M anager Group Leader Technical R eview er _ ... A d op tion D ate: 03/01/99 R e v is io n D a te : Z 1101 D ate O ^/eild D ate - of. o( D ate 1 .0 S c o p e a n d A p p l i c a t i o n __________________________________________________________________ 1.1 S cop e: T his m ethod describes the analysis o f serum extracts for fluorochem ical surfactants using H PLC -electrospray/m ass spectrom etry. 1.2 A p p lica b le C o m p o u n d s: Fluorochem ical surfactants or other fluorinated com pounds, or other ionizable com pounds. 1 .3 M a t r ic e s : R ab b it, rat, b o v in e , m o n k e y , and hum an seru m , or o th e r flu id s as d e sig n a te d in ,1 e validation report. 0014 W ord 6/95 3M Environmental Laboratori' ETS-8-5.2 Analysis of Serum Extract Us.ng ES/MS Page 1 ' U Page 46 3M Medical Department Study: T-6295.25 418-018:PAGE 1-420 A nalytical Report: FA C T -T O X -169 LR N -E01-0180 2 .0 S u m m a r y o f M e t h o d __________________________________________________________________________ 2.1 A lthough supported by a validation for m ost com m only used m atrices, this is a perform ance-based m ethod. C areful attention should be paid to m ethod Q C as there is great variability in sera. T his m ethod describes the analysis o f fluorochem ical surfactants extracted from serum or other fluids, using H PLC -electrospray/m ass spectrom etry, or similar system as appropriate. T he analysis is perform ed by m onitoring a single ion characteristic o f a particular fluorochem ical, such as the perfluorooctanesulfonate (PFO S) anion, m /z= 49 9 . A dditionally, sam ples m ay be analyzed using a tandem m ass spectrom eter to further verify the identity o f a com pound by detecting daughter ion s o f the parent ion. 3 .0 D e f i n i t i o n s ____________________________________________________________________________________ 3.1 A tm o sp h e r ic P ressu re Io n iza tio n (A P I): T he M icrom ass Q uattro II and U ltim a triple quadrupole system s allow for various m ethods o f ionization by utilizing various sources, probes, and interfaces. T h ese include but are not lim ited to: E lectrospray Ionization (E S I), A tm ospheric Pressure chem ical Ionization (A PcI), Therm ospray, etc. T he ionization process in th ese tech n iq u es occu rs at atm ospheric pressure (i.e., not under a vacuum ). 3 .2 E le c tr o sp r a y Io n iz a tio n (E S , E S I): a m ethod o f ionization perform ed at atm ospheric pressure, w hereby ions in solu tion are transferred to the gas phase via tiny charged droplets. T hese charged droplets are produced by the application o f a strong electrical field. 3.3 M ass S pectrom etry, M ass Spectrom eter (M S), T andem M ass S p ectrom eter (M S /M S ): T he A P I Q uattro II and U ltim a triple quadrupole system s are equipped with quadrupole m ass selective detectors. Ions are selectively discrim inated by m ass to charge ratio (m /z) and subsequently detected. A single M S m ay be em ployed for ion detection or a series (M S /M S ) for m ore specific fragm entation inform ation. 3 .4 C o n v en tio n a l vs. Z -sp ra y p ro b e in terface: T he latest m odels o f M icrom ass triple quadrupole system s (p ost 1998) utilize a "Z-spray" conform ation. T he spray em itted from a probe is orth ogon al to the c o n e aperture. In the conventional co n fo itn a tio n it is aim ed directly at th e c o n e aperture, after p a ssin g through a tortu ou s p ath w a y in th e co u n ter electrod e. T h ou gh the configuration is different, the m ethods o f operation, clean in g, and m aintenance are the sam e. H ow ever, Z-spray com ponents and conventional com ponents are not com patible w ith o n e another, but only w ith similar system s (i.e., Z -spray com p on en ts are com patible w ith som e other Z-spray system s, etc.) 3 .5 M a ss L yn x S oftw are: System softw are designed for the specific operation o f these Q uattro II triple quadrupole system s. Currently M assL ynx has W in d ow s 9 5 and W in d ow sN T 4 .0 versions. A ll versions are similar. For m ore details see the m anual sp ecific to the instrum ent (M icrom ass Q uattro II or U ltim a triple quadrupole M assL ynx or M assL ynx N T U ser's G uide). 4 .0 W a r n i n g s a n d C a u t i o n s _____________________________________________________________ 4.1 H ealth a n d S afety W arn in gs: 4.1 .1 U se caution w ith the voltage cables for the probe. W hen engaged, the probe em ploys a voltage o f approxim ately 500 0 V olts. 001478 W ord 6/95 5M Environmental Laboratori' ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 2 of 11 Page 47 3M Medical Department Study: T-6295.25 418-018:PAGE 1-421 Analytical Report: FACT-TOX-169 LRN-E01-0180 4 .1 .2 W hen handling sam ples o r solvents w ear appropriate protective g lo v es, eyew ear, and clothing. 4.2 Cautions: 4 .2 .1 D o not operate solvent pum ps above capacity o f 4 0 0 bar (5 8 0 0 p si) back pressure. I f the back pressure exceed s 4 0 0 bar, the H PLC w ill initiate autom atic shutdow n. 4 .2 .2 D o not run solvent pum ps to dryness. 5.0 In t e r f e r e n c e s ________________________________________________________________________________ 5.1 T o m inim ize interferences w hen analyzing sam ples, T eflon should not be used for sam ple sto ra g e or any part o f instrum entation that co m es in con tact w ith th e sam ple o r extract. 6 .0 E q u i p m e n t ___________________________________________________________________________ _________ 6.1 E quipm ent listed b elo w m ay b e m odified in order to optim ize the system . D o cu m en t any m odifications in the raw data as m ethod deviations. 6 .1 .1 6.1 .2 M icrom ass Q uattro II or U ltim a triple quadrupole M ass Spectrom eter equipped w ith an electrospray ionization source H P 1100 or A gilent low pulse solvent pum ping system , solvent degasser, colum n com partm ent, and autosam pler 7.0 S u p p l i e s a n d M a t e r i a l s ____________________________________________________________________ 7.1 Supplies 7 .1 .1 H igh purity grade nitrogen g a s regulated to approxim ately 1 0 0 p si (H o u se air or nitrogen system ) 7 .1 .2 H PL C analytical colum n, specifics to be determ ined by the analyst and docum ented in the raw data. 7 .1 .3 Capped autovials and capped 15 m L centrifuge tubes 8 .0 R e a g e n t s a n d S t a n d a r d s ___________________________________________________________________ 8.1 Reagents 8 .1 .1 M ethanol, H PLC grade or equivalent 8 .1 .2 M illi-Q TM w ater, all w ater u sed in this m ethod should b e M illi-Q TM waver or equivalent, and m ay be provided by a M illi-Q TO C Plus system or other vendor 8 .1 .3 A m m onium acetate, reagent grade or equivalent 8.2 Standards 8 .2 .1 T y p ica lly t w o m e th o d b lanks, tw o m atrix blanks, and eigh teer, m atrix standards :ie prepared during the extraction procedure. S ee E T S -8-4.2. SM Em ?rjumental Laboratory' ETS-8-5.2 Analysis of Serum Extract Using ES/MS 0 0 1 4 '; 9 Page 1 of 11 P o t'? 4 8 3M M edical D epartm ent Study: T -6295.25 418-018:PA GE 1-422 A n alytical Report: F A C T -T O X -169 L R N -E01-0180 9 .0 S a m p l e H a n d l i n g ______________________________________________________________________________ 9 .1 Fresh m atrix standards are prepared w ith each analysis. Extracted standards and sam ples are stored in cap p ed autovials or capped 15 m L centrifuge tubes until analysis. 9 .2 I f analysis w ill be delayed, extracted standards and sam ples can be refrigerated at approxim ately 4 C , or at room tem perature, until analysis can be perform ed. 1 0 .0 Q u a l i t y C o n t r o l ____________________________________________________________________________ 10.1 S o lv en t B lan k s, M eth o d B la n k s a n d M atrix B lan k s 1 0 .1 .1 S olven t blanks, m ethod blanks and matrix blanks are prepared and analyzed at least o n ce during the co u rse o f the study to determ ine if contam ination occurred during sam ple prep. 1 0 .1 .2 A nalyze at least o n e solvent blank prior to each calibration curve. 1 0 .1 .3 M atrix blanks should be analyzed w ith each sam ple list that includes undiluted extracts. 10.2 M atrix S p ik es 1 0 .2 .1 I f cu rves and m eth od Q C are prepared in a surrogate m atrix (e .g . cu rves in rabbit sera, sam ples are m onkey sera), matrix spikes and m atrix spike duplicates are prepared in blank sam ple m atrix (e.g ., m onkey sera) and analyzed to verify extraction efficiency. 1 0 .2 .2 I f cu rv es and m eth o d Q C are prepared in th e sam e m atrix as sam p les, n o additional m atrix spikes are required. 10.3 C on tin u in g C alib ration V erification s (C C V s) 1 0 .3 .1 C ontinuing calibration verifications are analyzed to verify the continued accuracy o f the calibration curve. 1 0 .3 .2 A nalyze tw o calibration standards (on e at each o f 2 levels) after every on e to ten sam ples, w ith a m inim um o f tw o per batch and alw ays finishing an injection sequence w ith at least tw o calibration standards. 1 1 .0 C a l i b r a t i o n a n d S t a n d a r d i z a t i o n ______________________________________________________ 11.1 A nalyze the extracted m atrix calibration standards prior to each set o f extracts. T he curve w ill b e p lotted by linear regression, w eigh ted 1/x, not forced through zero, using M assL ynx or other suitable softw are. 1 1.2 If the curve d oes not m eet requirem ents, perform routine m aintenance, reextract sam ples, or reanalyze the standard curve. 1 1 .3 For purposes o f accuracy w hen quantitating levels o f analyte at the lim its o f the curve range, it m ay b e n ecessa ry to u se either the lo w end or th e h igh en d o f th e calibration curve rather than the full range o f the standard curve. Exam ple: w hen attem pting to quantitate ap p roxim ately 10 p p b o f analyte, it m ay b e beneficial to generate a calibration cu rve con sistin g o f the standards from 5 ppb to 100 ppb rather than the full range o f th e curve (5 ppb to 1000 ppb). T his w ill reduce inaccuracy attributed to linear regression w eighting o f 0014S0 SM Environmental Laboratory' ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 4 of 11 Page 49 3M Medical Department Study: T-6295.25 418-018:PAGE 1-423 A nalytical Report: F A C T -T O X -169 LR N -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 d on e, no m ore than on e point should be u sed in com m on betw een the curves. F or exam ple, the low curve m ay include the follow in g points: 1, 5, 2 5 , 100, 250 ppb and the high curve m ay include the points: 2 5 0 , 500, 7 5 0 , 1000, 1250 ppb. 12.0 P r o c e d u r e s ________________ __ ___________________________________________________________ 12.1 A cq u isitio n S e t u p 1 2 .1 .1 On the M assL ynx main p age, set up a sam ple list nam e. S ave the list as instrum ent designator letter, last 2 digits o f test year-m o-day, and a letter that w ill increase through the alphabet w ith each additional list for that day. E xam ple Sam ple List: IY Y M M D D a or D 0 1 0 7 1 2 a I=instrum ent nam e (D for "D avey") Y Y =year o f test (01) M M =m onth o f test (07) D D =day o f test (12) a=first sam ple list (run) o f the day (the next sam ple list w ill end w ith %' th e n e x t `c ' and s o o n .) 1 2 .1 .2 A ssign a filenam e using the instrum ent designator letter, the last 2 digits o f year-m oday, and a 3 -d ig it sequential file num ber that starts w ith 1 suid in creases b y o n e for each filenam e. E xam ple File Nam e: IY Y M M D D ### or D 010712001 I=instrum ent nam e (D for "D avey") Y Y =year o f test (01) M M =m onth o f test (07) D D =day o f test (12) # # # = 3-d igit sequential file number starting w ith 1 through 9 9 9 (0 0 1 ) A lso, as part o f the sam ple list, assign a m ethod (M S) for acquiring, an inlet file, a bottle num ber, an injection volum e and sam ple descriptioni;. 1 2 .1 .3 T o create a m ethod, click on M ethod Editor button in the M S Status Pane and select SIR (Single Ion R ecording) or M R M (M ultiple R eaction M onitoring). Set Ionization M od e as appropriate and m ass to 4 9 9 or other appropriate m asses. A lso set the acquisition start and stop tim es. Save acquisition m ethod. I f M S/M S instrum ents are em ployed, additional product ion fragm entation inform ation m ay be collected. S ee M icrom ass M assLynx G U ID E TO D A T A A C Q U ISIT IO N for additional inform ation and M R M . 1 2 .1 .4 T ypically the analytical batch run sequence begins w ith solvent blanks and a set o f extracted matrix standards. 1 2 .1 .5 Sam ple extracts are analyzed w ith tw o C C V s injected every o n e to ten sam ples. S olven t blanks should be analyzed periodically to m onitor possible analyte carryover and are not considered sam ple extracts but m ay be included as such 001481 5M Environmental Laboratory ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 5 of 11 Page 50 3M Medical Department Study: T-6295.25 418-018:PAGE 1-424 A n alytical Report: FA C T -T O X 169 LR N -E01-0180 12.2 U sin g th e H P L C 1 2 .2 .1 S e t up sam p le tray accord in g to th e sam ple list prepared in S e c tio n 1 2 .1 .1 . 1 2 .2 .2 S et-up the H PL C to the follow ing conditions or at conditions th e analyst considers appropriate for optim al response. R ecord actual conditions in the instrum ent lo g b o o k : 1 2 .2 .2 .1 Sam ple size = 10 jiL injection 1 2 .2 .2 .2 Inject/sam ple = 1 1 2 .2 .2 .3 C ycle tim e = 10.0 m inutes 1 2 .2 .2 .4 F low rate = 3 0 0 pL/m in 1 2 .2 .2 .5 M obile Phase (program) Tim e 0 .0 0 min. 1.00 min. 5 .5 0 m in. 7 .5 0 m in. 8 .0 0 m in. M eOH 10% 10% 95% 95% 10% 2 .0 m M A m m onium a c e ta te (in H 20 ) 90% 90% 5% 5% 90% 12.3 Instrument Set-up 12.3.1 R efer to E T S -9 -2 4 for m ore details. 1 2 .3 .2 C h eck th e so lv en t level in reservoirs and refill if necessary. 12-3.3 C heck the stainless steel capillary at the end o f the probe. U se an ey ep iece to check the tip. T h e tip should be flat w ith no jagged ed ges. If the tip is found to be unsatisfactory, disassem ble the probe and replace the stainless steel capillary. The probe should be checked w eekly. 1 2 .3 .4 S et H P L C pum p to "O n". S et the flow to 10 - 5 0 0 pIVm in or as appropriate O bserve droplets com ing out o f the tip o f the probe. 1 2 .3 .5 Turn on the nitrogen. A fine m ist should be expelled w ith no nitrogen leaking around the tip o f the probe. Readjust the tip o f the probe if no m ist is ob seiv ed . 1 2 .3 .6 T he instrum ent uses these param eters at the follow in g settings. T h ese settings m ay change in order to op tim ize the response: 1 2 .3 .6 .1 D rying gas 2 5 0 -4 0 0 liiers/hour 1 2 .3 .6 .2 E S I nebulizing gas 10-15 liters/hour 1 2 .3 .6 .3 H PLC constant flow m ode, flow rate 10 - 5 0 0 pL/m in 1 2 .3 .6 .4 Pressure < 4 0 0 bar (T his param eter is not set, it is a guide to ensure the H PL C is operating correctly.) 1 2 3 . 7 C a r e fu lly g u id e th e p r o b e in to th e o p e n in g . In sert p r o b e until it w ill n o t g o a n y further. C onnect the voltage cables to the probe. 001482 3M Environmental Laboratory- ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 6 of 11 Page 51 3M Medical Department Study: T-6295.25 418-018:PAGE 1-425 A nalytical Report: FA C T -T O X -169 LR N -E01-0180 1 2 .3 .8 Print the tune p age, M S file, H PL C param eters, sam ple list and the M icrosoft W ord sum m ary p a g e and store in the study binder with a c o p y taped into the instrum ent log. 1 2 .3 .9 C lick o n start button in the M assL ynx m ain p age (this m ay vary am ong M assL ynx versions, see appropriate M assL ynx U S E R 'S G U ID E ). Ensure start and end sam ple num ber includes all sam ples to be analyzed. 13.0 D a t a A n a l y s i s a n d C a l c u l a t i o n s _____________________________________ 13.1 Calculations: 13 .1 .1 C alculate m atrix spike percent recoveries using the follow in g equation: __ Observed Result-M atrixB lank Result , % R ecovery = ------------------------------------------------------- X 100 Spiking Level 1 3 .1 .2 C alculate percent difference using the follow ing equation: % Difference Expected Cone. --Calculated Cone. x 100 Expected Cone. 1 3 .1 .4 C alculate actual concentration o f PFO S, or other fluorochem ical, in m atrix (pg/m L): ( Cone, o f PFOS Calc, from Std. Curve (ng / mL) x Dilution Factor) ( Initial Volum e o f M atrix (m L) + m L o f Surrogate Standard) Final Volume ( mL) 1 pg 1000 si 1 4 .0 M e t h o d P e r f o r m a n c e ______________________________________________________________________ 14.1 M ethod D etection Lim it (M D L ) and Limit o f Q uantitation (L O Q ) are m ethod, analyte, and matrix specific. Please see E T S -8 -4 .2 , A tta ch m en t B , for a listing o f current validated M D L and LO Q values. 14.2 S olven t B lan k s, M eth o d B lan k s, an d M atrix B lanks 14 .2 .1 Solven t blanks, m ethod blanks, and matrix blanks values m ust be b elow the low est active standard in the calibration curve. 14.3 Calibration Curves 14 .3 .1 T he coefficien t o f determ ination value for the calibration curve m ust be 0 .9 9 0 or better. SM Environmental Laboratorv ETS-8-5.2 Analysis of Serum Extract Using ES/MS 01483 Page 7 of 11 Page 5s 3M Medical Department Study: T-6295.25 418-018:PAGE 1-426 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 1423.2 AD activ e calibration cu rv e points m ust be w ithin 25% o f th e th eo retical value w ith the exception o f the L O Q point, which may deviate up to 30% . 14.3.3 C alibration standards w ith peak areas less than tw o tim es the curve m atrix blank m ust be deactivated to disqualify a data range that m ay b e affected by background levels o f the analyte. 14.3.4 L ow o r high curve points m ay be deactivated to optim ize a linear range appropriate to the data. 14.3.5 N o t including low o r high points dropped to optim ize the linear range, curve points may be deactivated if they deviate m ore than 25% from the theoretical value w hen the curve is evaluated o v er a linear range appropriate to th e data. 1423.6 O ne point below th e L O Q m ay rem ain active even if it deviates m ore than 30% or has a p eak area less than tw o tim es the m atrix blank; how ever, the L O Q wUl be defined at the low est point w ith acceptable deviation. 14.3.7 A valid calibration curve m ust contain at least 5 active points above and including the LOQ. 14.4 M a trix S pik es 14.4.1 T he average m atrix spike percent recoveries should be w ithin 30% o f the spiked concentration. R ecoveries outside o f this range should be discussed in the report. 14.5 C o n tin u in g C a lib ra tio n V erifications (C C V s) 14.5.1 C ontinuing calibration sam ples within the Unear range o f the run m ust show a percen t reco v ery w ithin 25% o f the spiked concentration. I f a C C V is o u tsid e o f this recovery, subsequent data should not be accepted. A cceptable data m ust be bracketed by the curve and passing CCVs. 1 4 .6 I f c rite ria liste d in th is m e th o d p e rfo rm a n c e se c tio n isn 't m e t, m a in te n a n c e m a y b e perform ed on the system and sam ples reanalyzed or other actions as determ ined by the an aly st. D o c u m e n t aU a c tio n s in th e a p p ro p ria te lo g b o o k . 4.7 If data are to be reported w hen perform ance criteria have not been m et, the data m ust be fo o tn o ted o n tables and discussed in the text o f the report. 15.0 Pollution Prevention and Waste Management__________________________ 15.1 Sam ple ex tract w aste and flam m able solvent is disposed in high B T U containers, and glass p ip ette w aste is d isp o sed in b ro k en glass containers located in th e laboratory. 16.0 Records______________________________________________________________ 16.1 T he first page o f each data packet generated for a study m ust have the follow ing inform ation included eith er in the header, in the fo o ter o r hand w ritten on th e page: study or project num ber, instrum ent, sample m atrix and tim e point, date, and analyst. 16.2 A data packet includes the following: data review sum m ary form , M assLynx quantify com pound sum m ary report for each target analyte, quantify calibration report fo r each targ et analyte (curve), m eth o d rep o rt, W o rd docum ent Usting set-u p p aram eters, tune 001484 5M Envii aumentai Laboratori- ETS-8-5.2 Analysis of Serum Extract Using ES/MS Page 8 o 11 Page 53 3M Medical Department Study: T-6295.25 418-018:PAGE 1-427 Analytical Repon: FACT-TOX-169 LRN-E01-0180 m ethod rep o rt, M assL ynx scanning m ethod report, H PL C m ethod rep o rt, sam ple list, and quantity sam ple report (chrom atogram ). 16.3 For each analysis, after printing the tune m ethod report, W ord docum ent listing, set-up param eters, M assLynx scanning m ethod report, and sam ple list, copy and tap e into the instrum ent runlog. T h e original is m aintained in the d a ta packet. 16.4 O n e a c h p a g e o f th e q u a n tify c o m p o u n d re p o rt, q u an tify c a lib ra tio n re p o r t (c u rv e ), an d quantify sam ple report (chrom atogram ), the following inform ation m ust be included either in th e h ead er, th e fo o ter, o r h an d w ritten on th e page: stu d y o r p ro ject num ber, instrum ent, m ethod, calibration (the m ethod and calibration are usually assigned the sam e nam e), analyst, and date. 16.5 T h e analyst m ust d ate and initial th e first page in a packet as long as their initials and date are electronically included on each page. If initials and date are n o : electronically included, they m ust date and initial each page. 16.6 Sum m arize d ata using suitable softw are (e.g., Excel) for inclusion in the final report. See Attachment A fo r an ex am p le o f a sum m ary sp read sh eet. 16.7 B ack up electronic data to appropriate m edium R ecord the file nam e and location o f backup electronic d ata in instrum ent log book. 1 7 .0 T a b l e s . D i a g r a m s . F l o w c h a r t s , a n d V a l i d a t i o n D a t a ______________________________ 17.1 A ttachm ent A: D ata sum m ary spreadsheet. 1 8 .0 R e f e r e n c e s __________________________________________________________________________________ 18.1 F A C T -M -4 .1, "E x tractio n o f P o tassiu m P erfluorooctanesulfonate o r O th er F luorochem ical com pounds from Serum for Analysis Using H PLC -Electrospray/M ass Spectrom etry 18.2 E T S-9-24.0, "O peration and M aintenance o f the M icrom ass A tm ospheric Pressure Ionization/M ass Spectrom eter Q uattro II triple quadrupole System s" 18.3 The validation report associated with this method is ETS-8-4.0 & 5.0-V-l. 1 9 .0 A f f e c t e d D o c u m e n t s ______________________________________________________________________ 19.1 E T S -8 -4 .2 , "E x tractio n o f P o tassiu m P erfluorooctanesulfonate o r O ther F luorochem ical C om pounds from S eram fo r Analysis U sing H PLC -Electrospray/M ass Spectrom etry" 'M Environmental Laboratorx ETS-8-5.2 Analysis of Serum Extract Using ES/MS 001485 Page 9 '1 Page. 4 3M Medical Department Study: T-6295.25 418-018:PAGE 1-428 A nalytical R eport: F A C T -T O X -169 L R N -E 01-0180 20.0 Revisions_____________________________________________________________ Revision N um ber. v1 Reason For Revision Section 6.1.2 Clarification o f H P 1100 system com ponents. S ection 11.1 A v erage o f tw o curves, n o t standard values, ai e u sed fo r plotting linear regression and added the 1/x w eighting o f th e curve. Section 12.2.2.4 Clarification o f solvent ram p. S ection 17.1 C hanged fro m attachm ent B to A R ev isio n D ate 04/02/99 2 10.1 Clarified when blanks are run. 10.2 Added instructions for when surrogate matrix is used. 10.3 Specify requirem ents 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 A llow 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 o f 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 o f matrix spikes. 14.5 Describes evaluation and use of CCVs. 14.5.! Evaluation o f CCVs. 16.0 Add requirements for records and documentation. Attachment A: Summary Spreadsheet tiTS-8-5.2 Analysis of Sei .un Extract Using ES/MS A/ Environmental Laboratori' 001486 Page 10 ' l l P a i' 55 3M Medical Department Study: T-6295.25 418-018:PAGE 1-429 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 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 o f Extraction/Analyst: Date of Analysis/Analyst: G roup D ose Sam ple# C oncentration up/m l. Initial VoL mL D ilution F a cto r F inal C one. ue/m L SloDe: Taken from linear regression equation. Groun/Dose: Taken from the study folder. Sample#: Taken from the study folder. Concentration (uc/m L): Taken from the MassLvnx inteeration summary. Initial Volum e (mL): Taken from the study folder. Dilution Factor: Taken from the study folder. Final Cone. fue/m L): Calculated by dividing the initial volume from the concem m ion Attachment A: Summary Spreadsheet ETS-8-5.2 Analysis of Serum Extract Using ES/MS S.U Environmental Laboratory 001487 Pi, 11 of 1 Page 5b 418-018:PA GE 1-430 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 Appendix D: Data Summary Tables A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Table 7. T O X 169 Data Summary of PFOS Concentration--Rat Serum (pg/mL) Tbnepolnt Female FD Gestation Day 21 Pupa F l Gestation Day 21 Female FO Lactation Day 5 Pups F I Lactation Day 5 Group Group 1 0.5% Tween 80 Group 2 mevalonic add Group 3 1.6m g/kg PFOS + mevalonic ad d 0.00706 0.00280 (n -6 ) 0.0183 0.0234 MS) 55.8 8.87 (n -6 ) <LOQ* (n=6) 0.0782 0.0836 (n=6) 207 25.6 (n=6) 0.0164 0.00869 (n -6 ) 12.7 27.4 (n -6 ) 127 24.7 (n -6 ) 02299 0.0196 (n -6 ) 3 3 3 3.14 (n -6 ) 1 2 4 1 8 .7 (n=3) Group 4 2 mg/kg PFOS + mevalonic add 82.4 22.6 (n=6) 251 55.9 (n=6) 189 4 8 2 (n=6) N/A Group 5 c h o le s te ro l Group 6 1.6 mg/kg PFO S + ch o le s te ro l 0.0251 0.00396 (n=6) 79.9 155 (n=6) 0.0828 0.0315 (n=6) 154 3 2 2 (n=6) 0.688 1.62 (n -6 ) 104 14.6 (n -6 ) 0.188 0.199 (n -6 ) 109 7.85 (n=2) Group 7 2 mg/kg PFOS + ch o le s te ro l 371 214 (n -6 ) 166 50.4 (n -6 ) 145 7.59 (n*6) 155 (n=1) G roups 0.4 mg/kg PFOS N/A N/A 2 7 2 18.7 3 6 2 3.84 (n=6) (n -6 ) G roups 0.8 mg/kg PFOS N/A N/A 42.6 6.92 53.1 27.7 (n-6) (n -6 ) Group 10 1 mg/kg PFOS G rtxp 11 1.2 m g /kg P FO S Group 12 1.6 mg/kg PFOS N /A N/A 142 24.5 (n=6) N/A N/A 142 332 (n -6 ) 5 2 3 * 26.0 (n -6 ) 86.0 102 (n=6) 169 32.1 (n -6 ) 84,4 17.5 (n -6 ) 147 162 (n -6 ) N/A Group 13 2 mg/kg PFOS 125 19.8 (n -6 ) 170 38.1 (n -6 ) 134 26.6 (n=6) 138 (n=1) 'LO O -- U m lt of Q uantitation = 0.00494 pg/m L N/A-- Not applicable NOTE: Results are expressed as group averages the standard deviation associated with the group. NOTE: It is not possible to verify tare recovery o f endogenous analyte from tissues w ithout radiolabeled reference m rtertal. The ori.y m easurement of accuracy available at this tim e, extracted calibration curves and m ulti-level com 'nuing calibration verifications (GCVs), Lidieate that *ha sera data are quantitative to 25% . 001488 -\\I Environmental Laboratory' Page 57 3M M edical D epartm ent Study: T-6295.25 3M Medical Department Study: T-6295.25 Appendix E: Data Spreadsheets 418-018:PAGE 1-431 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 3M Environmental Laboratorv 001489 Page 58 418-018:PAGE 1-432 3M Medical Department Study: T-6295.25 f a c t -to x -169 Analytical Report: FACT-TOX-169 Study: Efll-0180 Product NumtaarfTMt Substance): Maui*: M cdkod/Rrvtoott Analytical Equipment $y** Number Inu rnment Soitwam/Vcniop: R k e a m it R-Squand Value: Slope: Y-buercepc Dates of Exmetan)Analyst Dates of AmlysU/Aaalyst: Date of Data Reduction/Analyse S unplt D>U R AT SERA F0GD21 Group Dom Sample# Creep 1 0.5% Tween 80 Convoi C reep! M tvrieei Acid Conimi CreepJ 1.6 m|/k* PFOS Mevalonic And Creep 4 2 mpfc| PFOS Mcvmkec Acid Creeps Ordeero) Control Creupd i6m/k{PFOS Cholesterol Creep 7 2 m p tg PFOS * Cholesterol Group 11 1.6 ms/kf PFOS C reep 13 2mpAf PFOS rfOS . PcrfluonncuneMlfmic 10901F I09Q2F 10903F 10WUF* I0905F 10907F 10915F 10916FIQyiTF KJ918F 10919F 10920F IU930F" 10931F 10932F 10933F 10934F 10935F* 1Q94SF 10946F 10947F* 10949F 10950F 1095IF H1958F 109S9F 10960F* 10961F* 1U962F* 10963F* 10974F' 10975F 10976F 10978F 10980F IU985F KW87F* 109MF I0989F 09W tow tp nostiF* 1104SF* IKM6F JKM7F I1050F i I05JF tI0S4F 1I056F 11U58F 1lOftOF 11062F* PArT.Tnv.i4fl A rpA ii.nu Na Rat Sen EJS-M.2* ETS-S-32 Soup (80199. Amelia 062499 Masa)ytix3.4 See Below See Attachments See Attachment! See Attachment! 02/1S1-02/201 RWW/K1S a2/27l.03/15fll.(Q/2M>l MMH CQai1.0Vll.QV77A)L MMH Bo i #01-005AE Argus 418-018 buttai Vd. m. 0.70 050 072 0.45 0.45 073 060 04 0.55 0.73 053 0.60 0.30 OJO 055 063 0.69 036 OTO 071 aio 0.61 0.6S 0.55 0.38 oso 0?7 0.29 043 047 0.45 0.38 053 0.55 052 0.72 0.71 0.2 064 osi 0.43 MV 0.39 0.43 040 0 65 a 56 0.63 064 n .t 0.70 0.75 058 043 THPFOS PFOS PFOS niCM M Cnm ndM Verified DUottoa Coac. of PIT* Fader n t/m L trfifin L v rfH U e NA 1 836 S0I0227020 0.0119 NA 1 2.10 S010227G21 <LOQ (0.0W94 g|fiaL) NA ! 6.03 9010227022 0.00 a * NA 1 2.25 SDK827Q23 <LOQ (0.00)94 uffraU n a 1 3.73 9010327024 <LOQ (0-00194 ug/mL) NA 1 5.26 9010227025 unni na 1 3 91 S01Q227Q29 0 . 55 NA 1 5.97 S0I02270Q0 0.0124 NA 1 6J4 smaz2703i O.OU3 NA 1 4.45 S01Q227CQ2 <LOQ (0.00)94 ufftaL) NA 1 4J1 9010227033 <LOQ (0.00)94 u|fiaL) NA 1 634 S0I0227034 0.0106 NA 400 53.0 A0103I5020 707 NA 40) 69J A0103150Q1 555 NA 401 75.0 A010315Q22 545 Confirmed Low 400 69.0 A0LQ315023 431 NA 400 88.9 ADIOS15024 515 Confirmed Low 400 519 A010315025 5*7 Confirmed Low 1000 619 A01Q315029 89* Confirmed Low 1000 38.2 A01Q315030 539 Confirmed Low 1000 7.15 A01Q315031 715 Confirmed Low Confirmed Low Confirmed Low 1000 1000 1000 73.9 A01O315O32 4U AOI0315033 46.2 ADI0915034 121 74 3 839 NA 1 19.2 S0KQ27035 00-30 NA l 11.8 S01Q227036 ao35 NA 1 9.16 S010227037 0.014 NA 1 6.59 SOI0227O38 0K27 NA 1 978 A01Q3150IS no : 27 NA 11.2 Af>l0315019 no:38 Confirmed Low 1000 44.4 60103150 987 Confirmed Low 1000 24.3 6010315036 63 9 Confirmed Low 1000 51.9 A10315037 980 Confirmed Low toa 43.8 A010315Q38 796 Confirmed Low inno 316 A01Q31S042 74 2 Confirmed Low lOUD 46.7 A010315043 648 Confirmed Low 1000 71.6 A010315044 ICI Confirmed Low 1000 87. J A010315045 436 Confirmed Low 1000 718 A010315046 123 NA 5000 64.6 A010326016 633 NA 5000 646 60103260!? 513 NA 5000 5.J A nim arm i 41V Confirmed Low CcNtfinnud Low ini) JOCO 7ao 6010315050 64.7 60)0315031 179 ISO Confirmed Low Confirmed Low Confirmed Low Confirmed Low Kino icm laoo lonn 61.5 60I031055 7 60103)5056 75.5 6010315057 78.7 A0103150S8 154 no 115 125 Confirmed Low KUO 74.2 A0103ISS9 Confirmed Low imo 92.0 60103)5060 116 114 Confirmed Low 1000 77.8 60103)5061 III Confirmed Low 1000 84.3 6010315062 112 Confirmed Low imo 77.8 A010313063 12* Confirmed Low MOO 69J 6010315064 141 Esum.udcoiKejumjnfuutnmlvojuroetelowO.SmL. LAC02/2201 L R N -E 01-0180 m Rlg Mcaa p ro s (ta t RSD Sid. Dm. MS/MSDRPD 0.07706 00183 396 orm o 128 15.9 55* 8.87 27.4 82.4 226 0.0251 15.8 000396 194 79.9 13 5 57.8 371 214 J7.2 142 24.5 15.9 125 198 Date Entered/By: Date Verified/ By nMH/Ul.U3/22G1 LAC/03/28/ni KJH rn/nMiihoj 5.M Environmental Laboraron 001490 Page 59 418-018:PAGE 1-433 3M Medical Department Study: T-6295.25 ___ Analytical Report: FACT-TOX-169 r n v i * i W A*i7 Areus 418*018 LRN-EO 1 0 1 8 0 Sbriy: 01*0110 FACT-TOX-169. Arfus 41X-01I, Oee Generauon Reprodocoo Suidy of FFOS-Mcvatoruc AcxVOwteserol Challenge and NOEL Invtnifaiiou ie Rais Pmduci NumberfTcsi S a h n w .): NA Matrix: Rat San Method/Revision: Analytical Equipment System Humber. E S + 4 5 A E 5 -X -5 .2 Soup 020199. Amelia 06249S Instrument Software/Vcrsion: Massiyax 3.4 FUcname See Below R*SquMd Value Slope See Attachments See Attachments Y-Ixtereept Dates of Extraction/Analyit: See Attachments 02/15/01-02/2001 RWW/KJS Dries of Anaiysii/Aanlyst: 02/21/01,03/20RH.Q3/2M)I MMH Dale of Data Radaetum/Analyse 031)1/01.03/21/01,(0/291/0! MMH Sample Data Box oi-oosa-E RAT SERA PUPS F l CD21_________________ Group Dm Semple # laridel Vd. mL THPFOS Vcvism PFOS DUudou Factor PFOS Cone nt/m L Filename or m s /mL o r 4 Bee Mean p ro s n t/m L BSD Sid. Dr . MS/MSDBFO Group 1 05% Twuee 10 Cnntml G roup! Mevalonic Acid Coflttnl Group! 1.6 mg/kf PFOS Mevalonic Acid Group 4 2mg/kgFPOS Mevalonic Acid Group S Cholesterol Control 10901?" 10902?" I0903P* 10904P" 10905P* I0907P* 10915?" 10916?" 10917?" 1091IP* 10919?" 10920P* 10930P* 10931P" 10932?" 10933P" 10934?" 10935?" 1094SP* 10946?" 10947?" 10949?" IU950P* 10951?" 10954?" 10939?" 10960?" 10961?" 10962?" 0.14 ana 0.13 aos a is 0.06 a24 0.10 0.1X 0.06 aw 0.05 0.19 053 .22 0.15 a 19 0.21 0.1ft a i6 0.13 om 050 023 0.10 0.21 0.11 02 0.21 NA NA NA NA NA NA NA NA NA NA NA NA NA NA 2nd analysts OK 2nd analysis OK NA NA 2nd analysis OK Confirmed Low Confirmed Low Confirmed Low 2nd analysis OK 2nd analysis OK NA NA NA NA NA 1 196 SI0222112 <LOQ(ano4i|fiDU 1 0470 S01022XI13 <LOQ (0.00494 ep/mL) 1 4 20 S01Q22S114 <LOQ (000491 ag/mL) 1 a m S01Q22X1I5 <LOQ (000491 epfinL) 1 3.75 3)10224116 <LOQ<a004McAnL} 1 IT? 1022X117 LOQ <000491 vtAnL) 1 40.4 1022X121 0161 1 150 1022X122 <LOQ (0.00491 eg/mL) 1 19.6 1022X123 A.109 1 3.62 1022X124 <LOQ (0.00494 egfinL) 1 159 1022X125 0.177 1 a m 10221126 LOO (0.00494 etfinU 4D 1U2 A01Q320016 214 400 99.1 A10320017 172 400 120 AO1002X016 219 400 9a7 A01032X017 242 400 102 A010320020 214 400 95.4 A0IQ320Q21 in 400 119 A01032X01I 265 4(0 905 A010320TS3 226 400 X3.3 A0I0320024 256 400 ( .4 A010320025 345 400 131 A0KB2M19 175 400 137 A01092X020 239 1 14.0 10221177 o.un 1 115 S01022S124 0.0541' 1 9.IX 1022X129 0.089): 1 2a i 1022X130 0074:: 1 23.1 1022X134 anx2/> <LOO 007*2 207 251 NA NA 107 0.0836 037 25.6 225 55.9 34.0 I0963P* 0.31 NA 1 17.1 10221135 0.055:; 0.0822 0.0315 Group 6 1.6 mg/kg PFOS Cholesterol 10972P" I0974P* I097P* 0.13 0.26 0.00 Confirmed Low Confirmed Low Confirmed Low 400 455 A010320Q31 140 400 85.2 A01Q320Q32 :3i 400 44.0 A0(032tfZ33 (96 10976?" 10974P" 1098OP* 0.16 0.16 0.07 Confirmed Low Confirmed Low Confirmed Low 400 573* A010320Q34 145 400 445 A010320Q35 121 400 33.9 AU10320036 194 20.9 154 32.2 Group 7 2 mg/kg P ro s Cholesterol 10985P" 109*7?" 109X4?" 109X9?" 10990?" ,099IP- 0.24 0.IX 0.21 0.06 0 14 0.24 Confirmed Low Confirmed Low NA Confirmed Low Confirmed Low Confirmid Lew 400 755 A01032 137 126 400 75 Ani032003l ISO 400 M l A010320042 162 400 39.6 AD10320043 264 4 5S.4 Afi10320044 1SX 400 au.2 AUitnsmss l>4 304 166 80 4 Group 11 1Amg/kg PFOS Group 13 2 mg/kf PFOS lUMOP" 11044P- 11045P* 11046?" 11047?" 11050?" 11053?" t 1054?" 110S6P* II05X?* 11060?" I062P" 0.08 0 IX ais 023 0 11 0.21 OU 0.05 004 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 4UI 395 AM032I1M6 400 57.1 A0J0320D47 400 47.7 AI0320044 4U0 69Jt A0I0320049 400 46.4 AIQ320W 400 62.X A10320051 4UI , 41.7 Afii03211)55 400 ! 24.4 AO10320)36 4on 1 39.4 A010320157 400 i 915 AO1OJ20054 400 ! 595 A010320Q59 d ! 887 A010320060 196 127 127 112 169 120 .52 227 197 126 140 177 23.4 142 332 22.4 170 34.1 TFOS PerfluorraKuncM.onuie * Esoouted coneeetnmona. Jiioal volume bclax .p.5 iuL- LAC02/22AM Date Entcred/By: D*te Verified/ By: 034J7/01. 03/23/01 LAC/043/0I KJH 04/U6/01 ho, 3M Environmental Laboratory' 0 l4 9 i ; ge 60 418-018:PAGE 1-434 3M M edical D epartm ent Study: T-6295.25 r Stef?: EDIOIK> Marta: tH y d c il^ a p m M S f-- NaateT Siepe: FACT.TOX-149. AryM416-0)1, OnaG NA R a te i C7344.2 * ET34-U AttAi 06249L Snap020199 MaaMyv 3.4 Sm M ov FACT-T0X-K9 A rgus 418-018 A nalytical Report: FA CT-TO X -169 l l f m* MOWLl*'-- m i r a a t o L R N -E 01-0180 Dai of Canato^AaalyK Dai of Aaymiatfm: Datcdf Dan Madwuee/Aaaly* SaapkData RAT SERA FOLDS CaaMp Sm pitl Da trap 1 0 5 * Tam SOCantei IQM6F* 10909F 109UF* 11301P* I1SOOF H303F* Cnap 2 Montate AcMCmtei 10922F* I0923P" H315F* HSItfP* I1517F* (im p) IAnlfcFFOS I0931F MmlmieAdd 10M2P* II336F H337F* 11S30P* < m p4 U344F 2a/kg FP01 M e n te AcM II34SF II346P* I1J47P* U349F II330F* G iw pl I09MF* Cfaeteicnt Craami I0967P* I096CP* IIS5CF 1IS99F II36IF* Gmp * M B|fcf PPOS 109T9F T-Ila ri I09S3F* 1I572F* U373F 1IJ74F Gmp 1 2a*k|FFOS I0997F* Owlcatarl 1I5S6F MSfTF* llSttF* II SWF 10VWF 0.4 mffeg PPOS naooF* I1Q01F titaoF* 11602? 1160]F* Cnp 9 aSmf/ktFFOS 1I012F 1101IP* I16I2F II6I4F II617F Grvmp 10 lnV*|PTOS 1101VF* II021F* II023F* JI621F* 11624F* 1162SF* Grmp il 1.2mA| FFOS I103IF* II032F* II033F II&30F HA34F* I1639F* G m p 12 1.6 rap \| PFOS U04IF 11069F* II640F* 1I641F* IIM2F I1641F lim p U 2 m*A| PPOS II039F M654F 11*53F* 1165AF* 116J7F* SI65IF r r o s FentenmncunawanMi 02/130142/2001 RWW/KU OMliOI.OyiMJ.OS/2Ml.OMMl.aV3aOI MMH 030201.03/1901.03/7701.03/29A!.040201 MMH Sni 01-0QSA-E laal VaL L aw 062 0J9 0.4$ 0J2 0.36 QJJ 0.44 041 032 0.13 0.23 044 OSO 0.3 0.63 0.11 041 0JI 300 0.44 0.10 066 0.21 0.4 OI4 043 063 OJO 0.45 0J3 0J3 0J1 0J7 0J5 060 040 040 064 Q.4 0.40 100 0J3 0.47 OJS 039 063 0.37 0.64 0.S3 a a OSO 0.71 063 041 0.31 035 0J3 0.41 0 43 0.36 0.2? 0J3 0J2 021 013 0J9 0J2 Oil 0.12 in 0J6 0J3 0J2 OJl 041 01$ 0.72 FFOS pros FB aan VwtfM Factor C at a te i. NA 3 7.46 S0IO901020 NA 1 13.7 9010301021 NA 1 4.42 S0103010Z2 NA 1 14.0 S8J030I023 NA 1 M3 10301024 NA 1 3.25 S0I0301O23 NA > 27.6 A0I03140II NA $00 OJ A01Q32601V NA 1 6.64 A010316019 NA 20 4 0 A0103I6029 NA 20 154 A0103I6030 NA 20 27.3 A0103I6031 NA 1230 36.2 AO10326020 NA 1230 64.9 A010326031 NA 1230 31.7 A01Q324022 NA 1230 69.6 A01Q326023 M tejw O K NA 230 <2.6 A0JO32NB2 1230 36.1 A010326Q24 NA 1230 604 A0I032NQ3 NA 2300 7VJ A010326029 NA 1230 314 A010336030 2nd Mitya* OK NA NA 230 101 AOI032S0Z3 2300 42J A01032603I 1230 34.1 A010326032 NA 1 1M A01036090 NA 1 1.71 A0I031NS1 NA 1 10.9 A0I03160Z2 CeriRadffcfh 30 314 AKD16047 NA 1 29.6 A0I0316023 NA 1 to t A0IO316O24 NA 1230 49J A0I0326033 NA 2nd anatyaU OK 1230 33.0 A0IO326O34 230 124 A0I0321023 NA 1230 35.7 A0IO326O33 NA 1230 49.1 AO10326036 NA 1230 41.0 A0IQ326O37 NA 1230 46.0 A01032603I NA 1230 434 A0I0326O42 NA 1230 NU A0I0326O43 NA 1230 34.1 A0103260M NA 1230 44.4 A0I032NMS NA 1230 l i t A0I0326O46 2nd anitytu OK 2nd analyte OK 2nd Main* OK 1000 9.61 A0I032I029 1000 7.19 A01032S030 1000 10.4 A01032I03I 2nd Main* OK 1000 111 A010321032 2ndm in * OK 2nd n a ta ti OK 1000 16.3 A01032I033 1000 7.31 A01032I034 2miauiyuaOK Carinad law tea aaalpaMOJC 1DU0 23.) A01032I035 1000 21.2 A010316073 IODO n a AOIQMKM7 2nd Main* OK 2nd matyii OK 2nd anata OK 1000 36.0 A01032K3I 1000 40.2 A0IO330017 1000 21.6 A0I0330011 2ndMain* OK 2nd Mdytii OK 1000 LI AOI0330019 1000 19.1 AOI0330020 2nd a u ln * OK 2nd auyau OK NA 2rai analvala OK 1000 24.7 AOI033002I 1000 20.3 A010330022 30 162 A0I032NN7 1000 217 A0I0330023 2nd Main* OK 1000 34.0 A010330024 2nd analpi* OK 1000 262 AOIO330O23 ted a u ln * OK 1000 40.4 A0I0330026 2nd Main* OK 1000 466 A010330030 -2nd Main* OK 2nd mata OK 1000 24.9 AOI0330031 1000 9.20 A010330032 2im tn*O K 2nd a u ln * OK 2nd main* OK 2nd a u ln * OK 1000 12.4 AOI033003) 1000 62.3 A010330034 1000 261 A0I0330035 1000 24 1 AOI0930034 2nd a u ln * OK 2nd malva* OK IODO 120 A0I0330037 1000 99.5 A0I0330031 2nd ten * OK 2nd Main--OK 2ndMMyauOK 2000 37.7 A0I0330039 2000 411 A0IO33OO4S 2000 14.3 AOI03300M 2nd aulyaa OK 2000 224 A010330045 2nd analn* OK 2nd analnu OK 2000 11.1 A010330046 2000 53.7 A0I0330CM7 Eiwn--dotwmgMxia.al velara bao* 03 lid, LACO2/220I riarM tridaa adpms ateL ar* 9 0.0201 0.0220 <100 <000494 teteD ao3ii 00170 00146 00320 6L3 00133 2.71 2.42 2.31 103 162 102 134 113 146 131 199 167 270 161 207 0.0221 00122 00233 3.99 0.0392 00223 1)2 7V.6 100 i2l 11) 100 144 133 157 147 139 147 17.5 i6l 119 649 23.1 19.1 36.3 40.0 41. 43.0 51.5 34 2 313 310 >62 61.5 169 63 7 194 97.0 763 93.5 U.V 70* 140 193 in 201 120 (76 142 162 913 til 141 149 Da Intornia r Daa Vaaifiad/ iy. 030*01.03/2201 LAC/OV2M)t.O0VOl KJH O4060 M Maaa pros M *l 0.0114 12.7 127 IW 06U 104 143 27.2 42.6 52.3 SA0 169 134 KSD Sld. Dar. 47.2 27.4 19.3 24.7 41.2 233 1.62 140 14.6 7.J9 M7 6.92 491 10 2 190 311 19 9 26 1 001492 :\1 Environmental Laboratory Page Gl 418-018:PAGE 1-435 3M Medical Department Study: T-6295.25 FACT-TOX-169 Argos 418*018 A nalytical Report: FA CT-TOX-169 L R N -E 01-0180 Study: 01-0110 FACT-TOX-169. Arftu 41M ilt, One CeneneoA Reproduction Study of PTOS-Mevalonic Aad'Cfeokttuef m d NOEL tavciofuun m *'* Product NtmbcrfTc* Subatance): HA Rat Sen ETS4-4J & ETS-S^l Analytical Eqiapreet System Number Soup 020199 laaeument Softwaic/Veisien: M u ily n 3.4 See Below R-Squared Value: See Attachment! Slope See Aoicbeeali Y-lmcicqiu See Anifhraren D ata of Ejuiucuoo/Analyst QQ/I3A)MB/2(M)1 RWW/KJS D aw at Analyw/AMlyit: 03/084)1,03/21/01.03/2601,03/28/01.03/3001 MMH Date of Data Reduction/Aaalyst: OV13A)l.(CV22A)l.UV27A)l.03/2S/01.04ADAn MMH Sim ple D ltl Bos 0I-005A-E R A T S E R A P U T S F I L P S ___________________________ Creep Sample# InMal THPFOS PFOS pros Fllaeama Cencamttetiaw Date VoL VerlBM Piledwn Caoc efPFOS mL Feeler nf/mL e g f a L o r * Ree Creep 1 I0906F* 0.30 NA 1 11.0 5010308018 331366 A3V>Tween 80 Coeml 10909F* 1104 2nd Anelysit OK 1 Oil) A0MS21019 <LOQ(t 00494 og/mL) 1091 IP* (101 NA 1 000 SOKOOKBO <LOQ (T.00494 eg/reL) J150J/6F A93 NA 1 38.8 SDHOOttSl 10408 U302/7F 030 NA l 249 S010308022 10497 11303/tF AM NA 1 27.1 S01030MZ3 0.0423 Creep 2 10921F* 020 NA 1 204 SO1Q9Q80Q4 0.103 Mcrsiocuc A ad Cornel 10922F* 10923F* 0.31 006 NA 2nd Analysis OK 1 462 9010908023 <10214 1 l.7 | A010S21029 <LOQ (040494 ug/mL) 115IS/24F 1.00 NA 100 49.8 A010321036 4.98 1I316/21F* 043 NA 100 305 A01GQ21037 7.09 11SI7/20F 0.76 NA 100 33.7 AOicmicm 4.43 Creeps 1.6 e^/kg PFOS 109I9F* 10942F* U 10 aos NA NA 200 31.6 AO10321042 103 200 341 AD10S21O43 139 *>Mevalonic Acid 11536/37F* a 10 NA 200 64.7 A010Q21044 129 C reep ! 10964P* 0.13 NA 1 2.96 A01Q321000 <LOQ (000494 ug/mL) Cholesterol Control 10967F* I0969F* 023 aio NA NA l m 5010308034 0.0787 l 7.14 5010308033 0.0784 1133I/69F 066 NA 1 353 5010308036 1338 11339/7CF 1.00 NA 1 166 501(008037 1166 I1361/68F 1.00 NA 1 803 SO10301038 U0BQ5 C reep! I097VF* an NA 200 621 A0IQ321045 114 1.6 n r t t PFOS * ChoJ. 10983F* ai2 NA 200 61.8 A010321046 103 C reep 7 2 mg/kg 9 Choi. 10997F* 030 NA 500 93.3 A01Q32I047 135 C reu p l U4 eig/kg pros 10999F* noooF* an 023 NA NA 200 17.7 AO10926O48 311 200 48.4 AO10326049 42.1 110Q3F* 009 NA MO 17J AD103260S0 38.4 n60omp U602/9F* iifioydp- 047 028 0.17 NA NA NA 200 75.4 A01032105t 311 200 301 AOI0321035 35.8 200 312 AOI0926D51 36.7 Creep 9 0.8 mg/kg PFOS 11009F* 11012F* II013P* 0.06 013 007 Na NA na 200 18.9 A01U326035 18.9 200 332 A01U32103I 70.9 200 21.1 A0KD26036 21.1 I1612/13F 030 na 400 73.6 A01Q321060 58.9 II6I4/16F* 033 na 400 518 A010321061 AU4 UM7/I9F* 021 na 400 46.3 A010321062 88.2 C reep 19 now 0.2X na 1(11) 27.0 A01Q326Q57 962 1 rog/kg PTOS 11021F* 020 na inoo 13.0 A010326051 63.0 U023F* 0.26 NA mm 22.9 A01Q326O39 88.2 II621/22F* 1I624/29F* l liU/JHP 0-34 014 n*m NA NA NA 1000 310 ADI0326OA0 91.0 100 838 4010321(770 612 inun 733 AOicnannei ins C reep 11 1.2mg/kg PFOS 1103IF* IK02P* 008 A1A NA Na 100 382 A01Q330D48 477 a uno 105 AOKQ26063 U.4 Croup 13 2mg/kgPFOS I1CB3F* II036F* 11037F* | 1 1I634/39F* 1 J1US9F* 030 0 13 017 031 aw NA NA na na na 1000 28.1 A010326064 1000 9.03 A01032606I 1000 13.7 4010326069 inon 27.5 4010326070 :no .3 A01U326O71 MlO *12 ns <8.1 138 PFOS PmluoranctaoeauKonau Eiumated cone*nrraiiena. miual volume heiow0.5 reL 'A C 02/22/01 i > 4 pntiucunewasuaedfeuquinuunort KJHO4X>3/0l DatcEnvttMBy: 03/22/01.OiQJAN LAC/OiOXOI. 044001 JUH Dau Verified/ By: (M/06O1 ho) Mane PFOS ecAeL 0.0299 3.33 124 0.188 109 NA 36.2 33.1 844 .47 NA BSD Sid. Dev. MS/MSDRPD 66.2 0.0)98 94.4 3.14 13.1 18.7 106 MW 7.22 785 NA 10.6 3.14 521 27.7 2117 17.5 11 162 na 'M Environmental Laboratory 1493 Page 62 418-018:PAGE 1-436 3M Medical Department Study: T-6295.25 3M Medical Department Study: T-6295.25 A nalytical R eport: F A C T -T O X -169 L R N -E 01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 Appendix F: Example Calculations Formula Used for Sera Analyses in Study FACT TOX-169 AR (ng/m L ) x D F x F V (m L) x 1,0 p g = R ep o rted C o n cen tra tio n (p g /m L ) E V (m L) 1000 ng Calculation Used for Group 3, FO, GD21, Animal ID 10930F 53.0 ng/m L x 4 0 0 x 1.0 m L x 1.0 p g 0.3 mL 1000 ng = 70.7 pg/m L AR-- A nalytical result from M assLynx summary D F-- D ilution factor FV -- Final extract volum e (1 .0 m L unless otherw ise noted) EV -- V olum e o f sera extracted 3\{ Environmental Laboratori' 001494 Page 63 418-018:PAGE 1-437 3M M edical D epartm ent Study: T -6295.25 3M M edical D epartm ent Study: T-6295.25 A nalytical Report: FA C T-TO X 169 L R N -E 010180 Analytical Report: FACT TOX-169 LRN-E01-0180 Appendix G: Interim Certificate of Analysis -;-\I Eiwironme.-iol Laboiaton. 001495 Page 64 418-018:PAGE 1-438 3M Medical Department Study: T-6295.25 A nalytical Report: FA CT-TO X -169 L R N -E 01-0180 Centre Analytical Laboratories, Inc. 3048 R esearch Drive Phone: (814) 231-8032 S tate College, PA 16801 Fax: (814) 231-1253 or (814) 231-1580 ( INTERIM CERTIFICATE OF ANALYSIS Revision 1(9/7/00) C entre A n alytical L aboratories C O A R eference #: 023-018A 3M Product: P F O S ,L o t2 1 7 R eferen ce#: SD -018 ___________________________ P u r it y : 8 6 .9 % ___________________________ Test Name Specifications Result P u rity 1 86.9% A ppearance Identification NMR M etals (ICP/M S) 1. C alcium 2. M agnesium 3. Sodium 4. Potassium2 5. N ickel 6. Iron 7. M anganese Total % Im purity (NMR) Total % Impurity (LC/M S) Total % Impurity (GC/M S) Related Compounds POAA Residual Solvents (TGA) Purity by DSC Inorganic Anions (IC) 1. C hloride 2. Fluoride 3. Brom ide 4. Nitrate 5. N itrite 6. Phosphate 7. Sulfate* O rganic A cids s (1C) 1. T F A 2. PFPA 3. H FBA 4. NFPA Elem ental Analysis": 1. C arbon 2. Hydrogen 3. Nitrogen 4. Sulfur 5. Fluorine W hite Crystalline Powder 1. T heoretical V alue = 17.8% 2. Theoretical Value = 0% 3. Theoretical Value = 0% 4. Theoretical Value = 5.95% 5. Theoretical Value = 60% C onfoim s Positive 1. 0.005 w t/w t% 2. 0.001 w t/w t% 3. 1.439 w t/w t% 4. 6.849 w t/w t.% 5. <0.001 w t/w t.% 6. 0.005 w t/w t.% 7. <0.001 w t/w t.% 1.93 w t/w t.% 8.41 w t/w t% N one Detected 0.33 w t/w t.% N one Detected N ot Applicable1 1. <0.015 w t/w t.% 2. 0.59 wtVwt.% 3. <0.040 w t./w t% 4. <0.009 w t/w t.% 5. <0.006 w t/w t.% 6. <0.007 w t/w t.% 7. 8.76 w t/w t% 1. <0.1 w t/w t.% 2. <0.1 w t/w t.% 3. Q .10w t/w t% 4. 0.28 w t/w t% 1. 12.48 w t/w t.% 2. 0.244 w t/w t% 3. 1.74 w t/w t.% 4. 8.84 w t/w t.% 5. 54.1 w t/w t.% COA023-018A >M Environmental Laboratori' Page 1 o f3 Page 65 418-018:PAGE 1-439 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0180 Centre Analytical Laboratories, Inc. k 3048 R esearch Drive State College. PA 16801 ^ Phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Centre Analytical Laboratories COA Reference #: 023-018A Date o f Last Analysis: 08/31/00 Expiration Date: 08/31/01 Storage Conditions: Frozen <-I0C Re-assessment Date: 08/31/01 'Purity = 100% - (sum of metal impurities, 1.45% +LC/MS impurities, 8.41%+Inorganic Fluoride, 0.59%+NMR impurities, 1.93%+organic acid impurities, 0.38%+POAA, 0.33%) Total impurity from all tests = 13.09% Purity = 100% - 13.09% = 86.9% 2Potassium is expected in this salt form and is therefore not considered an impurity. 3Purity by DSC is generally not applicable to materials of low purity. No endotherm was observed for this sample. 4 Sulfur in the sample appears to be converted to SO4 and hence detected using the inorganic anion method conditions. The anion result agrees well with the sulfur determination in the elemental analysis, lending confidence to this interpretation. Based on the results, the SO4 is not considered an impurity. 5T FA HFBA NFPA PFPA Trifluoroacetic acid Heptafluorobutyric acid Nonofluoropentanoic acid Pentafluoropropanoic acid 6Theoretical value calculations based on the empirical formula, CeF i7 S0 3 "K+(MW=538) Thir work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). COA023-018A 3A1 Environmental Laboratori C 143' Page 2 ci Page 66 418-018:PAGE 1-440 3M Medical Department Study: T-6295.25 A nalytical Report: FA CT-TOX-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 en tre A nalytical L aboratories C O A R eference #: 023-018A LC /M S P urity Profile: Im parity 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. T he C5 value w as calculated using the average response factors from the C 4 and C 6 standard curves. Likew ise, the C7 value w as calculated using the average response factors from the C6 and C8 standard curves. 1. Prepared By: D avid S. B ell D ate Scientist, C entre A nalytical Laboratories // R eview ed By: / if /J o h n Flaherty v D ate Laboratory M anager, Centre A nalytical I.aboratories 1498 V C O A 023-018A 3M Environmental Laboratory Page 3 o f3 a g e 67 3M Me'dical Department Study: T-6295.25 3M M edical D epartm ent Study: T-6295.25 Appendix H: Report Signature Page 418-018:PAGE 1-441 A nalytical Report: FA CT-TO X 169 L R N -E 01-0180 Analytical Report: FACT TOX-169 LRN-E01-0180 (a Date Marvin T. Case, D.V.M., Ph.D., S p o n s o r R e p r e s e n ta tiv e Date Kristen J. Hansen, Ph.D., P rin c ip a l A n a ly tic a l In v e s tig a to r Date 100 I William K. Reagen, Ph.D., L a b o r a to ry M a n a g e r Date SM Environmental Laboraton- 001499 Page 68 418-018:PAGE 1-442 P F O S C oncen tration s in R at L ivers Sou th ern R esea rch Institute S tu d y A 3 4 4 .1.4 001500 418-018:PAGE 1-443 M ethods We first prepared control tissue homogenate by homogenizing rat liver from control rats 1:5 (by weight) with DI water. This was used to prepare the calibration standards. We spiked in known amounts o f test article (PFOS) and internal standard (PFOC) into aliquotes o f the blank tissue homogneate. A blank + int. std. and a blank - int. std were prepared with each calibration curve. The samples were also homogenized (1:5) by weight with DI water. Dilutions made to any o f these samples were with unspiked blank control liver homogenate prepared above. Then an equal amount o f internal stanard (PFOC) was add to each sample. Then to both the samples and standards we added 1 mL o f DI water, 1 mL o f 0.25/0.25 M carbonate/bicarbonate buffer, and 500 pL o f ion pairing reagent (0.5 M tetrabutylammonium hydroxide) adjusted to pH 10.5 with sodium hydroxide. The solution was mixed briefly and the we added 2.5 mL o f ethyl acetate. This mixture was shaken for 1 hr on a platform mixer. After an hour the samples are taken off and centrifuged at 2500 rpm for 5 min. The ethyl acetate layer is removed and evaporated to dryness under dry nitrogen. The residue is reconstitued and filtered through a syringe filter into autosampler vial for analysis. We then perform HPLC MS/MS using a MRM experiment on the samples and standards. 001501 Southern Research insitiute Study A344.1.4 PFOS in Rat Livers File Name Sample Name Measured Cone. (ng/mL) 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 FO 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 Calculated Cone, (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 *61,501 A PPE N D IX J H IST O PA T H O L O G Y REPO RT 001503 418-018:PAGE J-l RESEARCH PATHOLOGY SERVICES, INC. 4 3 8 E a st Butler A venue, N ew 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, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 SUBMITTED BY: _______________0__e_^_.___)_? . 2<rt> -iW. Ray Brown, D.V.M., Ph.D Veterinary Pathologist Decem ber 13, 2002 001504 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 Statement................................................................................... 5 001505 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 Numberof Animals 28 1" Dosage RODIH20 2'*' Dosage Vi hr +10 minutesafter 1* 0.5% Tween 60 3,dDosage 5 hr+30 minutesafter2nd RODI HjO 2 MevalonicAcid Control 3 1.6 mg/kg PFOS + MevalonicAdd 4 2 mg/kg PFOS + MevalonicAdd 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 MevalonicAdd 500 mg/kg MevalonicAdd 500 mg/kg MevalonicAdd 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/kgPFOS 0.8 mg/kg PFOS 500 mg/kg MevalonicAdd 500 mg/kg MevalonicAdd 500 mg/kg Mevalonic Acid NA NA NA NA NA 10 1mg/kg PFOS 20 NA 1mg/kg PFOS NA 11 1.2 mg/kg PFOS 20 NA 1.2 mg/kg PFOS NA 12 1.6 mg/kg PFOS 28 NA 1.6 mg/kg PFOS NA 13 2 mg/kg PFOS 28 NA 2 mg/kg PFOS NA The test and control articleswereconsidered 100% pureforthe purposeof dosagecalculations. Mevalonicacidwasdosedtwicedaily. The FO 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 001506 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 u te ro exposure) or via maternal milk during the lactation period. Dosages of the F0 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, ail raw data (remaining wet tissue, paraffin blocks, microscopic slides and histology records) will be returned to Argus Research Laboratories, Inc. for archiving. Rr -archPathology Sen-ices, Inc. 001507 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 n - within normal histologic limits. Research Pathology Services. Inc. -3 001508 418-018:PAGE J-6 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 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 PFOS 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. Research Pathology Services, 'nc. -4 001509 418-018:PAGE J-7 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC AC ID/C HOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS PROTOCOL 418-018 SPONSOR'S STUDY NUMBER: T-6295.25 HISTOPATHOLOGY REPORT QUALITY ASSURANCE UNIT STATEMENT Ail 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 Dates Reported to Date Reported Manaaement to Studv Directe, 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 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 i/LU.-D ^ J l r Karelh W. Harkins, BS Quality Assurance Unit . - ` 5 0 ? Date Research Pathology Sendees, Inc. A 001510 APPENDIX K HISTORICAL CONTROL DATA 001511 SUMMARYOF REPRODUCTIVE INDICES Crt:CD (SD)IGS B R RATS DAY21 C-SECTION 418-018:PAGE K-l PERIOD JUNE 1 998-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 33.4 (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 14.5 (13.6-15.9) AVERAGE # DEAD FETUSES 0.0 - AVERAGE # RESORPTIONS 0.5 (0.1-1.4) AVERAGE # EARLY RESORPTIONS 0.5 (0.1-1.4) AVERAGE # LATE RESORPTIONS 0.0 (0-0.1) 001512 418-018:PAGE K-2 SUMMARYOF REPRODUCTIVE INDICES Crl:CD(SD)IGS B R RATS DAY21 C-SECTION RANGE/STUDY MEAN or % 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) 001513 SUMMARYOF MATERWALNECROPSY OBSERVATIONS Crt:CD(SD)IGS B R RATS - DAY21 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 RANGE/ STUDY 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) 001514 418-018:PAGE K-4 SUMMARYOF FETALGROSS EXTERNALALTERATIONS Crt:CD (SD )lG S B R RATS - DAY21 C-SECTION 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 001515 418-018:PAGE K-5 SUMMARYOF FETALSOFTTISSUE ALTERATIONS Cr1:CD(SD)IGS B R RATS DAY21 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 UTTER INCIDENCE F FETAL INCIDENCE 001516 418-018:PAGE K-6 SUMMARYOF FETAL SKELETALALTERATIONS Cri:CD(SD)!GS B R RATS - DAY21 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 RANGE/ STUDY 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 001517 418-018:PAGE K-7 SUMMARYOF FETALSKELETALALTERATIONS Crl:CD(SD)IGS B R RATS - DAY21 C-SECTION ALTERATION MANUBRIUM Duplicated STERNEBRAE One or more incompletely ossified or not ossified Asymmetric Duplicated XIPHOID Duplicated HINDLIMB Extra digit present ' emur, bent RANGE/ STUDY 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) L: LITTER INCIDENCE F: FETAL INCIDENCE 00151 SUMMARY OF FETAL OSSIFICATION SITES SKELETAL AVERAGES Crl:CD(SD)IGS BR 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 .. (13.03-13.14) (5.85-5.97) - (7.11-7.63) (13.02-13.12) 1.00 3.99 1.00 .. (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) 001513 APPENDIX L STATEMENT OF THE STUDY DIRECTOR 001520 418-018:PAGE L-l Aigus 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 SPON SOR'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 Rule8, the Japanese M inistry o f Health and W elfare (MHW) Good Laboratory Practice Standardfo r Safety Studies on Drugsb and the Organisation for Econom ic Co-operation and D evelopm ent (OECD). The Revised OECD Principles o f Good Laboratory Practicesc 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 5 8 .130(c). The hard copy printout is considered the raw data. The rtability of the reference standard and internal standard were not determined. Argus Research Laboratories, Inc. a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. 1 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 Econom ic Cooperation and Development (1998). The Revised OECD Principles Good Laboratory Practices [C(97)186/Final]. 0015*1 APPENDIX M QUALITY ASSURANCE STATEMENT O Primedica 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 STATEM ENT A rgos Protocol: 418-018 Study D irector: Raymond G. York, Ph.D., DABT S p o n so r's S tu d y N u m b er: 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 M inistry of Health and W elfare (1997). Good Laboratory Practice Standardfo r Safety Studies on Drugs, MHW Ordinance Number 21, M arch 26, 1997. European Econom ic Community (1989). Council decision on 28 July 1989 on the acceptance by he European Economic Community o f an OECD decision/recommendation on compliance with principles o f good laboratory practice. Official Journal of the European Communities: Jegislation. 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 A UG 00 and 21 AUG 00. Vhe QAU inspection and report audit dates are listed below: Inspection Phase Inspection Datefs) Date(s) Findings Submitted to Study Director Date(s) Findings Submitted to Management Test Article Preparation Blood Collection Caesarean-Sectioning Tissue Collection Natural Delivery/Litter Oam/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 418-018:PAGE M-2 Observations Inspection Phase inspection Datefsl Date(s) Findings Submitted to Study D ire c to r__________ Date(s) Findings Submitted to Manaeement In-Life Raw Data Formulations Raw Data Report Tables Report Text Revised Report 8 JAN 01, 19 JAN 01, 22 JAN 01 to 76 JAN 01 and 79 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 78 FEB 01 25 FEB 01 to 26 FEB 01 and 23 M AR 01 to 79 M AR 01 i7 MAR 01, 30 M AR 01 and 02APR 01 to 03 APR 01 to 0 APR 01, 2 APR 01, 13 APR 01 and 16 APR 01 and 17 A PR 01 18 APR 01 04 MAY 01 and 07 M AY 01 17 JU L 01 iO OCT 01 30 OCT 01 . 1 JAN 02 04 FEB 02 27 MAR 02 10 DEC 02 01 FEB 01 19 FEB 01 01 M AR 01 29 M AR 01 03 A PR 01 16 APR 01 17 APR 01 18 APR 01 07 M AY 01 17 JU L 01 10 OCT 01 31 OCT 01 31 JAN 02 04 FEB 02 27 MAR 02 10 DEC 02 01 FEB 01 19 FEB 01 01 M AR 01 29 M AR 01 03 APR 01 16 APR 01 17 APR 01 18 APR 01 07 MAY 01 17 JU L 01 10 OCT 01 31 OCT 01 31 JAN 02 04 FEB 02 27 MAR 02 10 DEC 02 0o*S24 /x>x M atthew J. Vaneman, B.S. Date Manager, Regulatory Compliance ( .0 Cynthia M. Kelsch Quality Assurance Associate and Principal Auditor Date