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FINAL REPORT ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 FINAL REPORT DATE: 16 DECEMBER 2002 TITLE - ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACIDKHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS ARGUS STUDY NUMBER: 4 18-018 SPONSOR'S STUDY NUMBER: T-6295.25 TABLE OF CONTENTS SUBJECT PAGE I. SUMMARY AND CONCLUSION I- 1 A. Methods I- 1 B. Results 1-4 C. Conclusion 1-10 II. DESCRIPTION OF TEST PROCEDURES 11-1 A. Conduct of Study 11-1 B. Test Article Information 11-4 C. Control Articles Information 11-4 D. Vehicle Information 11-5 E. Test Article Preparation and Storage Conditions 11-6 F. Test System 11-7 G. Husbandry 11-9 H. Methods TI-1 1 m. RESULTS m- 1 A. Mortality m- 1 B. Clinical Observations m- 1 C. Necropsy Observations m-2 .. 11 ~- SUBJECT PAGE D. Terminal Body Weights and Organ Weights and Ratios of Organ Weights to Terminal Body Weights m-2 E. Body Weights and Body Weight Changes F. Absolute (g/day) and Relative (gkg/day) Feed Consumption Values m-3 m-5 G. Mating and Fertility m-7 H. Caesarean-Sectioning and Litter Observations m-7 I. Natural Delivery and Litter Observations m-7 J. Clinical Observations from Birth to Day 5 Postpartum and Necropsy Observations 111-8 K. Blood and Tissue Analyses m-9 L. Histopathology of Hearts and Thyroids Fl Generation Pups III-27 REFERENCES h1-28 APPENDIX A - REPORT FIGURES Figure 1. Body Weights - Fo Generation Female Rats - Groups 1,8 through 13 A- 1 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- 1 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-12 Table 4. Body Weights - Premating - Summary - Fo Generation Female Rats B-15 Table 5. Body Weight Changes - Premating - Summary - Fo Generation Female Rats B-18 ... 111 -- SUBJECT PAGE Table 6. Maternal Body Weights - Gestation - Summary - Fo Generation Female Rats B-2 1 Table 7. Maternal Body Weight Changes - Gestation - Summary Fo Generation Female Rats B-27 Table 8. Body Weights - Lactation - Summary - Fo Generation Female Rats Table 9. Body Weight Changes - Lactation - Summary - Fo Generation Female Rats B-30 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-5 1 Table 16. Mating and Fertility - Summary - Fo Generation Female Rats B-54 Table 17. Caesarean-SectioningObservations - Summary - Fo Generation Female Rats B-57 Table 18. Litter Observations (Caesarean-DeliveredFetuses) - 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 - F1 Generation Litters B-66 Table 2 1. Clinical Observations from Birth to Day 5 Postpartum - Summary - F1 Generation Pups B-75 iv SUBJECT PAGE Table 22. Necropsy Observations - Summary - F1 Generation Pups B-78 Table 23. Clinical Observations - Individual Data - Fo Generation Female Rats B-8 1 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 Table 3 1. Maternal Feed Consumption Values - Lactation - Individual Data - Fo Generation Female Rats B-200 B-2 13 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 - F1 Generation Litters B-26 1 V SUBJECT PAGE Table 38. Pup Vital Status and Sex from Birth to Day 5 Postpartum - Individual Data - F l Generation Pups B-274 Table 39. Clinical Observations from Birth to Day 5 Postpartum - Individual Data - F l Generation Pups B-287 Table 40. Necropsy Observations - Individual Data - F1 Generation Pups B-292 APPENDIX C - PROTOCOL AND AMENDMENTS C-1 to C-67 APPENDIX D - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY D-1 to D-6 APPENDIX E - CERTIFICATE OF ANALYSIS E-1 to E-2 APPENDIX F - HOMOGENEITY AND CONCENTRATION ANALYSIS F-1 to F-21 APPENDIX G - TEMPERATURE AND RELATIVE HUMIDITY REPORTS G-1 to G-5 APPENDIX H - BIOANALYTICAL REPORTS (ANILYTICS) H-1 to H-125 APPENDIX I - BIOANALYTICAL REPORTS (PRIMEDICA WORCESTER, SPONSOR AND SRI) 1-1 to 1-444 APPENDIX J - HISTOPATHOLOGY REPORT J-1 to J-7 APPENDIX K - HISTORICAL CONTROL DATA K-1 to K-8 APPENDIX L - STATEMENT OF THE STUDY DIRECTOR L- 1 APPENDIX M - QUALJTY ASSURANCE STATEMENT M-1 to M-2 vi 4 18-018:PAGE I-1 TITLE: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACIDKHOLESTEROL 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-administrationof mevalonic acid or cholesterol could prevent or antagonize the adverse maternal and fetal effects (reduced maternal body weight gain, increased incidence of stillborn, decreased birth weight and decreased pup survival) observed in a previous study with PFOS (Argus Research Protocol 418-008,3M Study Number T-6295.9) and to better define the noobserved-effect-level (NOEL). A. 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-sectioningon 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 (ROD1 H20). 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). The dosage groups and dosing schedule were as follows: 4 18-018:PAGE 1-2 Dosage Groups Number ~~~~~ 1 Identification Vehicle Control 2 3 14 Mevalonic Acid Control 1.6 mgkg PFOS + Mevalonic Acid 2 mg/kg PFOS + i Mevalonic Acid 5 Cholesterol Control 6 1.6 mg/kg PFOS + Cholesterol 7 2 mg/kg PFOS + Cholesterol 1st dosage RODI H20 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mdkg Mevalonic Acid 500 mg/kg Cholesterol 500 mg/kg Cholesterol 500 mg/kg Cholesterol Dosing Schedule 2nd dosage 1/2 hour rt 10 minutes after 1st dosage 0.5% Tween@ 80 0.5% Tween@80 I .6 mg/kg PFOS 2 mg/kg PFOS 0.5%Tween@ 80 3rd dosage 5 hr 2 30 minutes after 2nd dosage RODI H20 500 mg/kg Mevalonic Acid 500 mgkg Mevalonic Acid 500 mg/kg Mevalonic Acid Not applicable 1.6 mgkg PFOS Not applicable 2 mg/kg PFOS Not applicable The dosages, concentrations and dosage volumes were as follows: Compound I Dosage I Concentration I Dosage Volume I PFOS 1 PFOS I 1.6 PFOS 2 0.20 I 5 0.32 5 0.40 5 Dosages were adjusted for the most recently recorded body weight and given at approximately the same times each day. 418-018:PAGE 1-3 The rats were given the test article, control article and/or vehicle daily beginning 42 days before cohabitation (maximum of 14 days) and continuing through DG 20 (rats assigned to Caesarean-sectioning),DG 24 (rats assigned to natural delivery that did not deliver a litter) or day 4 of lactation (DL 4, rats that delivered a litter). F1 generation pups were not directly given the test article, control article and/or vehicle, but were possibly exposed to the test article or control articles during maternal gestation or via maternal milk during the lactation period. Rats were observed for viability at least twice each day of the study. Observations for clinical signs of effects of the test article and deaths were made daily prior to dosage administration, approximately one hour after the second dosage and at the end of the working day. These observations were also recorded on the day of sacrifice. Body weights were recorded weekly to cohabitation, daily during presumed gestation, on DL 1 (rats assigned to natural delivery) and at sacrifice. Feed consumption values were recorded weekly to cohabitation, on DGs 0,7, 14,21 and 25 (if necessary) and DLs 1 and 5 (rats assigned to natural delivery). Mating was evaluated daily during the cohabitation period and confirmed by observation of spermatozoa in a smear of the vaginal contents and/or a copulatory plug observed in situ. The rats assigned to natural delivery were observed for adverse clinical signs during parturition, duration of gestation and pup viability at birth. The rats were also observed for fertility index, gestation index, number of offspring per litter, number of implantation sites, general condition of the dam and litter during the postpartum period and viability index. Maternal behavior was recorded on DLs 1 and 5. Litters were examined after delivery to identify the number and sex of pups, stillborns, live births and gross external alterations. Each litter was evaluated for viability at least twice each day of the 5-day postpartum period. F1 generation pups in each litter were counted once daily. Physical signs were recorded each day. Pooled litter weights were recorded on DLs 1 (birth, liveborn 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 4 18-018:PAGE 1-4 maintained frozen. The median liver lobe was frozen. A section from the remaining portion of the liver was fixed in gluteraldehyde. The remaining portion of the liver was frozen. Fetuses were pooled by litter and litter body weights were recorded. Blood samples were collected from each fetus. The liver from each fetus was collected. One fetal liver from each litter was removed and fixed in gluteraldehyde. The remaining fetal livers in each litter were divided into three samples of equal number. Two of the samples were frozen. The remaining samples were flash frozen in liquid nitrogen and maintained frozen. On DL 5 the rats assigned to natural delivery were sacrificed and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Blood, milk, heart and liver samples were collected from maternal rats on DL 5. Following collection of milk and blood samples, the heart and liver of each dam were collected. The liver weight was recorded. A portion of each liver sample (right lateral lobe) was flash frozen and maintained frozen. The median liver lobe was stored frozen. The heart and a section from the remaining portion of the liver were fixed in gluteraldehyde. The remaining portion of the liver was frozen. On DL 5 , pups were sacrificed and examined for gross lesions. Necropsy included a single cross-section of the head at the level of the frontal-parietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Blood samples were collected from each pup. For Replicate 1, blood samples were pooled per litter, samples from male and female pups combined. For Replicate 2, blood samples were pooled (two litters per sample within each dosage group). The liver from each pup was collected, pooled (by sex per litter) and the pooled weight recorded. One liver from each litter pool was removed and fixed in gluteraldehyde. The remaining livers in each litter pool were divided into three samples of equal number. Two of the samples were stored frozen. The remaining sample was flash frozen and maintained frozen. The hearts were collected from the first two male and two female pups from each litter (pooled by sex per litter) and were fixed in gluteraldehyde. Thyroids were excised from each pup, pooled (by sex per litter) and retained. Microscopic examination was made of the heart and thyroid from one male and one female F1 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 mgkg PFOS + mevalonic acid) and 5 (cholesterol control) died as the result of intubation accidents or natural causes. All other rats survived to scheduled sacrifice. 4 18-018: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 2 1 were significantly reduced in Groups 6 and 7 (1.6 and 2 mgkg 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 mgkg PFOS alone) and in Groups 6 and 7 (1.6 and 2 mgkg 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 mgkg 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 m g k g 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 mgkg 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 m g k g PFOS alone) and Groups 6 and 7 (1.6 and 2 mgkg 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 mgkg. Body weight gains for the entire gestation period were significantly reduced only in Groups 6 and 7 (1.6 and 2 mgkg PFOS + cholesterol, respectively). Groups 3 and 4 (1.6 and 2 mgkg PFOS + mevalonic acid, respectively), Groups 6 and 7 ( I .6 and 2 mg/kg PFOS + cholesterol, respectively) and Groups 9, 11, 12 and 13 (0.8, 1.2, 1.6 and 2 mgkg 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 4 18-018:PAGE1-6 increase in body weight gain during the third week of gestation. Body weights were significantly reduced in Group 12 (1.6 mgkg 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 mgkg PFOS + mevalonic acid) on DGs 4 through 13, and in Group 4 (2 mgkg 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 mgkg 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 mgkg 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 mgkg 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 mgkg 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 mgkg 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 mgkg 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 mgkg 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 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 mgkg 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 mgkg PFOS + cholesterol, respectively) on DGs 0 to 7 , 7 to 14, and 0 to 2 1. 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 mgkg PFOS + cholesterol, respectively) on DGs 14 to 2 1. 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 mgkg 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 mgkg 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 mgkg 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 significantlyreduced in Groups 6 and 7 (1.6 and 2 mgkg 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-sectioningobservations on DG 21 were based on eight pregnant dams in each of Groups 1 through 7, 12 and 13. No Caesarean-sectioningor 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 mgkg 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 mgkg PFOS + cholesterol, respectively). The number of dams with all pups dying on DLs 1 to 5 was 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 significantlyincreased in Groups 12, 13,4,6 and 7. The numbers of surviving pups per litter were significantly reduced on DLs 2 , 3 , 4 and 5 in Groups 12, 13, 3,4, 6 and 7. A dosage-dependentpattern 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 mgkg PFOS + mevalonic acid, respectively), Groups 6 and 7 (1.6 and 2 mgkg 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 mgkg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mgkg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mgkg PFOS + cholesterol). Increased numbers of litters with pups that were cold to touch occurred in Groups 11, 12, 13,4, 6 and 7. The number of litters with pups not nursing was increased in Groups 12, 13, 3,4, 6 and 7; the increase was significant in Group 7. Necropsy observations in pups that were stillborn or found dead included increases in the numbers of pups with no milk in the stomach in Groups 9, 10, 11, 12 and 13 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively), Groups 3 and 4 (1.6 and 2 mgkg PFOS + mevalonic acid, respectively), and Groups 6 and 7 (1.6 and 2 mgkg PFOS + cholesterol, respectively). The incidence of this observation was significantly increased in Group 6. All other clinical and necropsy observations in the F1 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 mgkg 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 mgkg 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. 4 18-018: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 mgkg 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 significantlyincreased in DG 21 fetuses from dams administered 1.6 or 2 mgkg 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, I, 1.2, 1.6 or 2 mgkg PFOS. The glucose was significantlyincreased in the DL 5 dams that were treated with 2 mg/kg PFOS alone and triglycerides were significantlydecreased in lactating dams treated with 1.6 or 2 mgkg 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 mgkg PFOS were significantlyreduced 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 mgkg PFOS. Cholesterol in the blood and triglycerides were significantly decreased in lactating dams that were treated with cholesterol and 1.6 or 2 mgkg PFOS and glucose was significantly increased in lactating dams that were treated with cholesterol and 2 mgkg PFOS. Liver triglyceride levels were increased or significantly increased in DL 5 dams administered 1.6 or 2 mgkg 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 mgkg PFOS. Free T4 levels in the F1 generation pups from lactating dams administered0.4, 0.8, 1, 1.2, 1.6 or 2 mgkg PFOS were significantly reduced. Free T3 levels in pups from lactating dams administered 0.4,O.S or 1mgkg PFOS were significantly reduced, however, levels were significantly increased, in pups from lactating dams administered 1.6 mgkg PFOS. Total T4 levels in pups from lactating dams administered 0.4,0.8 or 1 mgkg 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 mgkg PFOS. Total T4 were significantly decreased in pups from lactating dams that were treated with mevalonic acid and 1.6 mgkg PFOS. LDL and HDL were significantly increased in the pups from 4 18-018:PAGE I-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 mgkg PFOS. Total T3 and free Tq were significantlydecreased 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 mgkg/day PFOS (Group 13). C. Conclusion On the basis of these data, it does not appear that coadministrationof 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 mgkg PFOS (the 0.8 mgkg 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 mgkg (dosages of 0.4 m g k g and higher caused decreased pup body weights when normalized for litter size). G e o r g h . Dearlove, Ph.D., DABT Date Associate Director of Research k&hond G. Y o r k m . , DABT Date Associate Director of Research and Study Director 418-018:PAGE 11-1 11. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study A.1. Sponsor 3M Corporate Toxicology, 3M Center, Building 220-2E-02, St. Paul, Minnesota 55 144-1000 A.2. Testing Facility Argus Research, 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 A.3. Study Number 4 18-018 A.4. Sponsor's Study Number T-6295.25 A S . 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)"' 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)'*', the Japanese Ministry of Health and Welfare (MHW)'3' and the European Economic Community (EEC)'4'. There were no deviations from the GLP regulations that affected the quality or integrity of the study. Quality Assurance Unit findings derived from the inspections during the conduct of this study are documented and have been provided to the Study Director and the Testing Facility Management. 4 18-018:PAGE11-2 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.lO. 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 4 18-018:PAGE 11-3 A.16. Dates of Technical Performance A.16.a. Replicate 1 Rat Arrival Dosage Period Rats Assigned to Caesarean-Sectioning (42 days before cohabitation and continuing through DGa 20) Rats Assigned to Natural Delivery [42 days before cohabitation and continuing through DG 24 (rats that did not deliver a litter) or DLb4 (rats that delivered a litter)] Cohabitation Period Male 1 (7 days maximum) Male 2 (7 days maximum) Scheduled Sacrifice (DG 2 1 Caesarean-sectioning) DG 25 Sacrifice No Confirmed Date of Mating Sacrifice Delivery Period (DL 1) Scheduled Sacrifice (DL 5) 22 AUG 00 29 AUG 00 - 02 NOV 00 29 AUG 00 - 18 NOV 00 09 OCT 00 PM - 16 OCT 00 AM 16 OCT 00 PM - 23 OCT 00 AM 31 OCT 00 - 03 NOV 00 05 NOV 00 - 09 NOV 00 17 NOV 00 31 OCT 00 - 15 NOV 00 04 NOV 00 - 19 NOV 00 A.16.b. Replicate 2 Rat Arrival Dosage Period Rats Assigned to Natural Delivery [42 days before cohabitation and continuing through DG 24 (rats that did not deliver a litter) or DL 4 (rats that delivered a litter)] Cohabitation Period Male 1 (7 days maximum) Male 2 (6 days maximum) DG 25 Sacrifice Delivery Period (DL 1) Scheduled Sacrifice (DL 5) 05 SEP 00 14 SEP 00 - 02 DEC 00 25 OCT 00 PM - 01 NOV 00 AM 01 NOV 00 PM - 07 NOV 00 AM 20 NOV 00 - 23 NOV 00 16 NOV 00 - 29 NOV 00 20 NOV 00 - 03 DEC 00 a. DG is an abbreviation for day of (presumed) gestation. b. DL is an abbreviation for day of lactatiodpostpartum. 4 18-018:PAGE11-4 A.17. Records Maintained The original report, raw data and reserve samples of each lot of the control article and vehicle components are retained in the archives of Argus Research. Any preserved tissues are retained in the archives of the Testing Facility for one year after mailing the draft final report, after which time the Sponsor will decide their final disposition. All unused prepared formulations were discarded at the Testing Facility. Unused bulk test article was returned to the Sponsor. B. Test Article Information B.l. Description Perfluorooctane Sulfonic Acid Potassium Salt (PFOS) - light-colored powder B.2. Lot Number 217 B.3. Date Received and Storage Conditions The test article was received on 25 August 2000, and stored at room temperature. B.4. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask and safety goggles or safety glasses and a face-shield) were taken when handling the test article. B.5. Analysis of Activity Information regarding the identity, composition, strength, and purity of the test article is on file with the Sponsor. The purity is 98.9%. A Certificate of Analysis is available in APPENDIX E. C. Control Articles Information C.1. Descriptions Mevalonic Acid - white with a yellow cast, semisolid Cholesterol - white powder C.2. Lot Numbers Mevalonic Acid - 070K26603 and 080K26 18 Cholesterol - 119H0218 418-018:PAGE 11-5 C.3. Dates Received and Storage Conditions The mevalonic acid was received from Sigma Chemical Company, St. Louis, Missouri, on 1 August 2000 (lot 070K26603) and 21 September 2000 (lot 080K2618), and stored frozen. The cholesterol was received from Sigma Chemical Company, St. Louis, Missouri, on 20 July 2000, and stored at room temperature. C.4. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask and safety goggles or safety glasses and a face-shield) were taken when handling the control articles. C.5. Analysis of Purity Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the control articles that would have interfered with the results of this study. The purity of the mevalonic acid is approximately 97%. The purity of the cholesterol is 95%. The expiration dates for the mevalonic acid are August 2004 (lot 070K26603) and September 2004 (lot 080K2618). The expiration date for the cholesterol is July 2004. D. Vehicle Information D.l. Description 0.5%Tween@80 prepared using Tween@80, a yellow liquid, and reverse osmosis membrane processed deionized water (RODI H20). Reverse osmosis membrane processed deionized water (RODI H20). 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. 4 18-018:PAGE 11-6 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.1. Sample Information Sample Type Date StoragelShipping Size Retained Conditions Shipped To Date Shipped Homogeneity" 28 AUG 00 29 AUG 00 5mL 27 SEP OOb Frozen (-20C) Sponsor 27 SEP 00 Concentration` 18 SEP 00 01 NOV 00 15 NOV 00 Frozen (-20C) Sponsor 18 SEP 00 01 NOV 00 15 NOV 00 Control Articles Reserve Mevalonic Acid Lot 070K26603 Lot 080K26 18 Cholesterol 01 SEP 00 30 OCT 00 01 SEP 00 Frozen Frozen Testing Facility 13 SEP 00 38 NOV 00 13 SEP 00 Vehicle Component Reserves Tween@ 80 ROD1 H20 01 SEP00 01 SEP 00 I Room temperature I Room temperature Testing I Facility I Archives 13 SEP 00 13 SEP 00 a. A syringe was used to withdraw samples from the top, middle and bottom of the h hest PFOS concentration on the first day of preparation. Each sample was divided into two aliquots and placed into glass containers, one of 2 mL and one of 3 mL. One aliquot (2 mL) was shipped for analysis; the other aliquot (3 mL) was retained at the Testing Facility as a backup sample. b. A larger preparation size was required for dosing; therefore, according to the Standard Operating Procedures of the Testing Facility, if preparation is 30% larger or smaller, homogeneity must be reanalyzed. C. A syringe was used to withdraw samples from each PFOS concentration once during each of the following periods of study: premating, gestation and postnatal. Each sample was divided into two aliquots, one of 2 mL and one of 3 mL, and placed into glass containers. One aliquot (2 mL) was shipped for analysis; the other aliquot (3 mL) was retained at the Testing Facility as a backup sample. 4 18-018:PAGE 11-7 E.2. Analytical Results Samples were sent to Exygen Research, State College, Pennsylvania, for analysis. PFOS levels in the Day 0 and 10 stability and concentration samples bracketing the range of concentrations and conditions of this study were determined. Analysis of samples on 14 February 2002 to 15 February 2002 showed an average percent recovery f standard deviation for PFOS in the dosing solutions as 93% rt 7% and ranged from 83% to 105%, when compared to the assigned concentration, which was not considered to be significant. Results of the stability and concentration analyses are available in APPENDIX F. F. Test System F.l. Species Rat F.2. Strain Crl:CD@(SD)IGS BR VAF/Plus@ F.3. Supplier (Source Charles River Laboratories, Inc., Raleigh, North Carolina F.4. Sex Female (Note: Male rats were used only for the purpose of breeding and are not considered part of the Test System). F.5. Rationale for Test System The Crl:CD@(SD)IGS BR VAF/Plus@rat was selected as the Test System because: 1) this strain of rat has been demonstrated to be sensitive to reproductive and developmental toxins and has been widely used throughout industry for reproductive and developmental toxicity evaluations; 2) historical data and experience exist at the Testing 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. 4 18-018:PAGE11-8 F.6. Test System Data Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (8)the Day after Arrivala Weight (g) at Study Assignment Replicate 1 180 27 JUN 00 57 days 167 - 204 196 - 224 Replicate 2 179 10 JUL 00 58 days 170 - 228 199 - 247 F.7. Breeder Male Rat Data Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) the Day after Arrival Weight (g) at Cohabitation - Replicate 1 Weight (g) at Cohabitation - Replicate 2 Shipment 1 150 29 F%B 00 71 days 245 - 339 473 - 767 452 - 773 Shipment 2 150 20 MAR 00 72 days 286 - 344 537 - 748 593 - 791 F.8. Method of Randomization Female rats were received in two shipments, separated by two weeks. The first shipment was designated as Replicate 1 and the second shipment was designated as Replicate 2. Upon arrival, the male and female rats were assigned to individual housing on the basis of computer-generatedrandom 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 Croups 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 Croups 1 to 7, 12 and 13 of Replicate 1 with a confirmed date of mating were assigned to Caesarean-sectioning on DG 2 1. 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 MonelB self-piercingear 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. 418-018:PAGE 11-9 unique permanent number. Cage tags were marked with the study number, permanent rat number, sex, test article identification and dosage level. Pups were not individually identified during lactation, all parameters were evaluated in terms of the litter. G. Husbandry G.l. Research Facility Registration USDA Registration No. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 et seq. 6.2. 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%". 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 Guidefor the Care and Use of Laboratory Animals@). 6.4. Lighting An automatically-controlledfluorescent light cycle was maintained at 12-hourslight: 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). 418-018:PAGE 11-10 G.6. Feed Rats were given ad libitum access to Certified Rodent Diet@#5002 (PMI Nutrition International, St. Louis, Missouri) in individual feeders. G.7. Feed Analysis Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration for certified feed or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the feed that would have interfered with the results of this study. 6.8. Water Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad libitum from an automatic watering system and/or individual water bottles attached to the cages. Chlorine was added to the processed water as a bacteriostat. G.9. Water Analysis The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, Pennsylvania) and monthly for possible bacterial contamination (Analytical Laboratories, Inc., Chalfont, Pennsylvania). Copies of the results of the water analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the water that would have interfered with the results of this study. G.10. Nesting Material Bed-o'cobsB bedding (The Andersons Industrial Products Group, Maumee, Ohio) was used as the nesting material. G . l l . 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. H. Methods H.l. Dosage Administration The Lasage groups and dosing scheule were as follows". 418-018:PAGE 11-11 I sage Groups Number I 1 Identification Vehicle Control 2 Mevalonic Acid Control 3 1.6 mgkg PFOS + Mevalonic Acid 4 2 mg/kg PFOS + Mevalonic Acid 5 Cholesterol Control 6 1.6 m g k g PFOS + Cholesterol 7 2 mg/kg PFOS + Cholesterol 0.4 mgkg PFOS Dosage Groum and Dosing Schedule Dosing Schedule I 2nd dosage I 3rd dosage t 500 mg/kg Mevalonic Acid 500 mg/kg Mevalonic Acid 0.5% Tween@80 0.5% Tween@80 1.6 m g k g PFOS 500 mgkg Mevalonic Acid 500 mg/kg Mevalonic Acid 500 mg/kg 2 mg/kg PFOS 500 mg/kg I I Mevalonic Acid 500 mg/kg Cholesterol 0.5% Tween@80 Mevalonic Acid Not Applicable 0.8 mgkg PFOS 1 m g k g PFOS 1.2 mgkg PFOS 12 1.6 mgkg PFOS 2 mg/kg PFOS a. See APPENDIX D, items 2 through 4. 4 18-018:PAGE 11-12 The dosages, concentrations and dosage volumes were as follows": Compound Identification Dosage (mdkdday 1 Concentration (mg/mL) Dosage Volume (mLW 1 0.5%TweenB 80 I 0 I 0 I 5 I Mevalonic Acid 1000 ' 100 5" Cholesterol 500 100 5 PFOS 0.4 0.08 5 PFOS 1 0.20 5 PFOS 1.2 0.24 5 I' T* he test and control Mevalonic acid articles were considered was dosed twice daily. 100% pure for the purpose of dosage calculations. The rats were assigned to groups as follows: Dosage Grow 1 Identification I Vehicle Control Mevalonic Acid 1.6 mg/kg PFOS + Mevalonic Acid 2 mg/kg PFOS + Mevalonic Acid 5 Cholesterol Control 1.6 mg/kg PFOS + Cholesterol 2 mg/kg PFOS + Cholesterol 8 0.4 mgkg PFOS 9 0.8 mg/kg PFOS 10 1 mg/kg PFOS 11 1.2 mg/kg PFOS Number 1 of Rats --28 Replicate 1 Assigned Rat Numbers Replicate 1 1 Replicate 2 10930-10935,10937, 10940 10929, 10936,10938, 10939. 10941. 10942 1529 1543 28 10945 -10947,10949 - 10943, 10944, 10948, 1544 - 1557 109.51.10953.10954 10952.10955. 10956 28 10958 - 10965 I 11558- 11571 10972, 10971,10973,10977, 28 10974 10976' 10978' 10979, 10983, 10984 1572 1s85 10980 - 10982 28 1100998875,- 10993 10986, 10994 - 10998 11586 - 11599 20 I 10999 - 11008 I 1~ 16~ 00-1.16.0.9 ~~ 20 Not applicable 20 Not applicable I 11009-11018 1 11019- 11028 11610- 11619 11620- 11629 20 Not applicable 11029- 11038 11630- 11639 1 1040, 11039, 11044- 11047, 11041 - 11043, 11048, 11640 - 11653 110.50 - 11052 11049 110s3' 11054'11056' 11058*11060' 11062' 11063. 11066 11055, 11057, 11059, 11061, 11064, 11065 11654- 11667 a. See APPENDIX D, items 5 through 7. 4 18-018: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 testkontrol article; such rats may not have received any or all of their daily dosage(s) on the day of parturition. F l 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 (inutero 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 perioda. Observations for clinical signs of effects of the test article and deaths were made a. See APPENDIX D, items 8 and 9. 418-018:PAGE 11-14 daily prior to dosage administration, approximately one hour after the second dosage and at the end of the working day (approximately one hour after the third dosage)a. 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 sacrificea. 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)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 born that survived five days). Maternal behavior was recorded on DLs 1 and 5'. Deviations from expected maternal behavior were recorded, when and if observed, on all other days of the postpartum period. H.5.b. F1 GenerationPups Litters were examined after delivery to identify the number and sex of pups, stillborns, 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. F1 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, liveborn 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. 418-018:PAGE II-15 H.6. Gross Necropsya H.6.a. Fo Generation Rats Assigned to Caesarean-Sectioning Eight rats from Groups 1 to 7, 12 and 13 (Replicate 1) were sacrificed by carbon dioxide asphyxiation on DG 2 1. Blood and liver samples were collected from these ratsb. The rats were Caesarean-sectionedand 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. Representativephotographs 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") 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 (5-20 "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 (5-70C). The median liver lobe was stored frozen (5-20C). 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 (I-2OoC). Fetuses were pooled by litter and litter weights were recordedd. a. A table of random units was used to select one control group rat assigned to Caesarean-sectioningfrom which all tissues examined at necropsy were retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions. b. See APPENDIX D, item 12. C. See APPENDIX D, item 13 d. See APPENDIX D, item 14. 4 18-018: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 (5-20C). 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 (I-20C). The remaining samples were flash frozen in liquid nitrogen and maintained frozen (I-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 (5-20C). 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 (520C). 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 ( ~ 7 0 C ) .The median a. See APPENDIX D, item 12. 4 18-018:PAGE 11-17 liver lobe was stored frozen (520C). 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 (approximately0.3 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 (520C). 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") 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 performeda. 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. Representativephotographs 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. F1 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 liveborn, 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. 4 18-01$:PAGE 11-I8 Sample tubes were labeled with the study number, rat identification,date of collection, study day, sample identification and collection timepoint. Blood samples were collected via cardiac puncture from each pup. For Replicate 1, blood samples were pooled per litter, samples from male and female pups combined. Approximately 1 mL of blood was transferred into serum separator tubes; any remaining blood was transferred into EDTA-coated tubes. The samples were spun in a centrifuge and the resulting serum was divided into two aliquots of approximately 200 mL and approximately 300 mL of serum, and stored frozen (5-70C). Any resulting plasma was transferred into one aliquot and stored frozen (5-70C). 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 (570C). Resulting plasma was transferred into one aliquot and stored frozen (5-70C). The liver from each pup was collected, pooled (by sex per litter) and the pooled weight recordeda. 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 (5-20C). The remaining sample was flash frozen in liquid nitrogen and maintained frozen (5-70C). 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. H.6.d. Sample Shipment and Analyses Sample Storage/Shipping Conditions Dams Assigned to Caesarean-Sectioning Serum - aliquot 1 4-20C Serum - aliquot 2 5-20"C Recipient Sponsor AniLytics Plasma Liver - right lateral lobe Liver - median lobe Liver - section Liver - remainder 4-20C <-70C I-20C Gluteraldehyde I-20C Primedica Worcester Sponsor Swonsor Sponsor AniLytics 4 18-018:PAGE 11-19 Analysis PFOS Total cholesterol, glucose, total & free T3, T4,TSH (Groups 1, 11, 12, 13), LDL, HDL, trigl ycerides" Mevalonic acid (Groups 1 , 2 , 3 , 4 , 11, 12 and 13) Possible biochemical PFO~~ S Possible EM LDL, HDL, total Pooled serum - aliquot 1 1-20"C Pooled serum - aliquot 2 <-20C Pooled Plasma Liver - I liver Liver - 1/3 pooled livers Liver - 1/3 pooled livers Liver - 1/3 pooled livers I-20C Gluteraldehyde <-7O"C I-2O0c 4-20C Sponsor AniLytics Primedica Worcester Sponsor Sponsor Sponsor AniLytics PFOS Total cholesterol, glucose, total & free T3, T4,TSH (Groups 1, 1I, 12, 13), LDL, HDL, trigl yceridesa Mevalonic acid (Groups 1, 2,3,4, 11, 12 and 13) Possible EM Possible biochemical PFOS LDL, HDL, total Milk Serum - aliquot 1 Serum - aliquot 2 4-20C 520C I-20C Plasma Liver - right lateral lobe Liver - median lobe Liver - section Liver - remainder Heart 220C 4-70C 2-20"C I-20C Gluteraldehyde AniLytics Sponsor AniLytics Primedica Worcester Sponsor Sponsor AniLytics Sponsor Total cholesterol PFOS Total cholesterol, glucose, total & free T3, T4,TSH (Groups I, 11, 12, 13), LDL, HDL, triglyceridesa Mevalonic acid (Groups 1, 2, 3,4, 11, 12 and 13) Possible biochemical PFOS -~Possible EM LDL, HDL, total cholesterol, triglycerides Possible EM and light microscopy I Sample Pups - Replicate 1 Serum - aliquot 1 Serum - aliquot 2 StorageIShipping Conditions I 5-70C I 5-70C Plasma Serum - aliquot I Serum - aliquot 2 5-70C S-70"C 5-70C Plasma <-70"C Liver - 1 liver per sex Liver - 1/3 pooled livers Liver - 1/3 uooled livers Liver - 113 pooled livers Gluteraldehyde I-70"C <-20c] <-2OoC Hearts - 2 males and 2 females per litter" Pooled Thyroidsd Gluteraldehyde 10% neutral buffered formalin 418-018:PAGE 11-20 Recipient Analysis Sponsor AniLytics Primedica Worcester Sponsor AniLytics Primedica Worcester Sponsor Sponsor Suonsor AniLytics Sponsor Testing Facility PFOSa Total cholesterol, glucose, total & free T3, T4,TSH (Groups 1, 11, 12, 13),LDL, HDL, trig1ycerides' Mevalonic acid (Groups 1, 2, 3,4, 11, 12 and 13) PFOS" Total cholesterol, glucose, total & free T3, T4,TSH (Groups 1, 11, 12, 13), LDL, HDL, triglycerides' Mevalonic acid (Groups 1 , 2 , 3 , 4 , 11, 12 and 13) Possible EM Possible biochemical PFOS LDL, HDL, total cholesterol, triglycerides Possible EM and light microscopy Possible histopathology Results of these analyses are available in APPENDICES H and I. H.7. Data Collection and Statistical Analyses Data generated during the course of this study were recorded either by hand or using the Primedica Argus Automated Data Collection and Management System and the Vivarium Temperature and Relative Humidity Monitoring System. All data were tabulated, summarized and/or statistically analyzed using the Primedica Argus Automuted 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). 4 18-018:PAGE 11-2J The following schematic represents the statistical analyses of the data: Type of Testa I. Parametric A. Bartlett's Testc 11. NonDarametricb A. Kruskal-Wallis Test (175% ties) Significant Not Significant Significant Not Significant Nonparametric Analysis of Variance I Dunn's Test 111. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution ..................... a. Statistically significant probabilities are reported as either ~ 5 0 . 0 5or ~50.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. 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.O, 1.2, 1.6 and 2.0 mgkg PFOS alone). Group 2 (mevalonic acid control) data were compared to Groups 3 and 4 (1.6 and 2.0 mgkg 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'"). 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 Variances(' and the Analysis of Variance(l2)w, hen appropriate [i.e., Bartlett's Test was not significant (p>O.OOl)]. If the Analysis of Variance was significant (p50.05),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 @lO.OOl)], the Kmskal-Wallis Test(14)was used, when less than or equal to 75% ties were present. In cases where the Kruskal-Wallis Test was statistically significant (p20.05),Dunn's Method of Multiple comparison^('^) was used to identify the statistical significance of the individual groups. If there were greater than 75% ties, Fisher's Exact Test('6)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('4). 4 18-018:PAGE111-1 111. 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 mgkg PFOS + mevalonic acid) and 5 (cholesterolcontrol) 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 (p10.01) in Group 5 (mevalonic acid control) as compared to Group 1 (vehicle control). These increases were attributed to an aversion to the taste of mevalonic acid; the incidence of these observations was comparable among the three mevalonic acid groups (Groups 2 , 3 and 4). All other clinical observations during the precohabitation, gestation and lactation periods were considered unrelated to the test or control articles because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in one to four rats in a group. These observations included localized alopecia, dental problems (missing, broken or misaligned incisors), chromodacryorrhea, scabs on paws and limbs, bent tail, missing or swollen digit, swollen snout, red perivaginal substance, lacrimation, scab on mouth, tip of tail missing, ptosis, soft or liquid feces, rales, red substance in cage, labored breathing, scant feces, swollen ear, dehydration, emaciation, decreased motor activity, pale in appearance, cold to touch, and ulceration or scab on chest or limb. The significant increase 418-018:PAGE 111-2 (p10.01) in the incidence of localized alopecia on the back during the gestation period in Group 11 (1.2 mgkg 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 mgkg 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-SectionedRats Terminal body weights of rats Caesarean-sectionedon DG 21 were significantlyreduced (p10.05 or p10.01) in Groups 6 and 7 (1.6 and 2 mgkg PFOS + cholesterol, respectively) as compared to Group 5 values (cholesterol control). The weight of the liver was significantly increased (p10.05) 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 (p10.05 or p10.01) 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 (p10.05)in Group 3 ( I .6 mgkg 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 (p10.05 or p10.01) 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 mgkg PFOS + cholesterol, respectively) as compared to Group 5 values (cholesterol control). The ratios of the liver weights to the terminal body weights were significantlyincreased (p10.05 or p10.01) in Groups 9, 11 and 13 (0.8,1.2 and 2 mgkg PFOS alone, respectively), compared to Group 1 (vehicle control), reflecting slight reductions in terminal body weights andor slight increases in liver weights in these groups as compared to the Group 1 values. The liver weight was significantly increased (p10.01) in Group 3 (1.6 mgkg PFOS 4 18-018:PAGE 111-3 + mevalonic acid) as compared to Group 2 (mevalonic acid control). The ratios of the liver weight to the terminal body weight were significantly increased ($50.05 or p50.01), 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 ($50.05 orp10.01) 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 ($10.05 orplO.O1) 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 mgkg PFOS alone) were significantly reduced (pl0.05) 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 (p10.01) 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 (p10.05) 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 mgkg. Significant increases or reductions (p10.05 or p50.01) in body weight gains in Group 8 (0.4 mgkg 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 (p10.05) only in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol). Groups 3 and 4 (1.6 and 2 mgkg 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 mgkg PFOS alone, respectively) had significantly reduced (p50.05 or p50.01) 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 4 18-018:PAGE 111-4 significant increase (p10.05) in body weight gain during the third week of gestation (DGs 14 to 21). Body weights were significantly reduced (p50.05 orp50.01) in Group 12 (1.6 mgkg 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 administere-dalone. Body weights were significantlyreduced (p50.05 or p10.01) in Group 3 (1.6 mgkg 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 significantlyreduced (p10.01) in Groups 6 and 7 (1.6 and 2 mgkg PFOS + cholesterol, respectively) on DGs 0 through 21, compared to Group 5 (cholesterol control). Gestation body weights were significantly increased (p10.05) 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 significantreductions 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 (p50.05 orplO.O1) 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 (p50.05)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 (p50.05)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 (p10.05or ~10.01i)n Groups 6 and 7 (1.6 and 2 mgkg PFOS + cholesterol, respectively),compared to Group 5 (cholesterol control), and a significant (p10.05) body weight loss occurred on DLs 1 to 5 in Group 7. 418-018:PAGE 111-5 F. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values (Summaries - Tables 10 through 15; Individual Data - Tables 29 through 31) F.1. Precohabitation Absolute and relative feed consumption values were significantlyreduced (p10.05 or p10.01) 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 (p10.05) 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 mgkg. Absolute feed consumption was significantlyincreased (p10.05) in Group 8 (0.4 mg/kg PFOS alone) on DSs 29 to 36; and absolute and relative feed consumption values were significantly increased (p10.05)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 (p10.05 or ~ 1 0 . 0 1i)n 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 ($10.05orp10.01) in Groups 3 and 4 on DSs 8 to 15, 15 to 22,22 to 29 and 36 to 42 (Group 4 only). This effect of PFOS and mevalonic acid was reflected in the significantly reduced (p10.05 or p10.01) 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 (p10.01) 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 (cholesterolcontrol). This effect of PFOS and cholesterol was reflected in the significantlyreduced (p10.05 or p10.01) absolute and relative feed consumption values for the entire premating period (DSs 1 to 42) in both groups. Relative feed consumption values on DSs 8 to 15, 15 to 22,22 to 29 and 1 to 42 were significantly increased (pS0.05 or p10.01) 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 (p10.01) on DSs 29 to 36 and absolute feed consumption was significantly increased (p10.05) on DSs 36 to 42 in the cholesterol control group (Group 5) as compared to the vehicle control group (Group 1). 418-018:PAGE 111-6 F.2. Gestation Absolute and relative feed consumption values were significantly reduced (p10.05 or p10.01) during the first week of gestation (DGs 0 to 7) in Groups 9 through 12 (0.8, 1, 1.2, 1.6 and 2 mg/kg PFOS alone, respectively). Absolute feed consumption continued to be significantly decreased (p10.05) in Group 13 on DGs 7 to 14, and relative feed consumption was significantly increased (p10.05) 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 (p10.05 orp10.01) 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 (p10.01) in Groups 3 and 4 (1.6 and 2 mgkg 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 (p10.05 or p10.01) in Groups 3 and 4 on DGs 0 to 7 and 14 to 21. When PFOS was administered along with cholesterol, effects on feed consumption values were also comparable to the effects of administration of PFOS alone. Absolute feed consumption values were significantly reduced (p10.01) in Groups 6 and 7 (1.6 and 2 mgkg 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 (p10.01) in Groups 6 and 7 on DGs 0 to 7 and significantly increased (p10.05 or p10.01) 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 (p10.01) in Groups 12 and 13 (1.6 and 2 mgkg PFOS alone, respectively). Relative feed consumption values for the same period were reduced in Groups 9 through 13, reaching statistical significance (p10.05 or ~ 1 0 . 0 1i)n Groups 9, 10, 11 and 12 (0.8, 1, 1.2 and 1.6 mgkg PFOS alone, respectively), compared to Group 1 (vehicle control) values. When PFOS was administered along with rnevalonic 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 (p10.01) in Groups 3 and 4 (1.6 and 2 mgkg 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 (p10.01) in Groups 6 and 7 (1.6 and 2 mg/kg PFOS + cholesterol, respectively) on DLs 1 to 5, as compared to Group 5 (cholesterol control). 418-018:PAGE 111-7 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 (p10.01) for Group 6 (1.6 mg/kg PFOS + cholesterol), as compared to Group 5 (cholesterolcontrol). 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-sectioningobservations on DG 2 1 were based on eight pregnant dams in each of Groups 1 through 7, 12 and 13. No Caesarean-sectioningor litter parameters were affected by dosages up to 2 mgkg 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 (p10.05 or ~ 5 0 . 0 1f)or Croup 13 (2 mgkg 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 (p10.05)in Group 7 (2 mgkg PFOS + cholesterol);although this value is above the range observed historically at this Testing Facility",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 (p10.05or ~ 1 0 . 0 1i)n Groups 9 through 13 (0.8, 1, 1.2, 1.6 and 2 mgkg 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 mgkg PFOS + cholesterol, respectively), as compared to the corresponding control a. See APPENDIX K (HISTORICAL CONTROL DATA). 4 18-018:PAGE 111-8 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 p10.01 in Groups 13,4 and 7. The pup viability index (number of live pups on DL 5 per number of liveborn pups on DL 1) was significantly reduced (p10.05 orplO.O1) 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 (p10.05 orp10.01) in Groups 12, 13,4, 6 and 7. The numbers of surviving pups per litter were significantly reduced (p10.05 orplO.O1) on DLs 2, 3 , 4 and 5 in Groups 12, 13,3,4,6 and 7. A dosage-dependentpattern 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 ( I .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 (p10.0 1) pup body weights on most weighing days (DLs 1 through 5). When average pup weight was normalized for litter size, Groups 3 and 4 (I .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 alone, respectively) were also significantly reduced (p10.05 or p10.01) 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 liveborn and stillborn pups were comparable across all 13 dosage groups. The number of dams with stillborn pups was significantly increased (p10.01) in Group 8 (0.8 mg/kg PFOS alone) and significantly reduced (p10.05) 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 (p10.01) 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 atplO.O1 in Group 7. 418-018:PAGE 111-9 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 (p10.01) in Group 6, as compared with Group 5 (cholesterol control). All other clinical and necropsy observations in the F1 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 (p10.01)in Groups 8,9, 10, 11, 12 and 13, reflecting the single occurrence of this observation in the vehicle control group (Group 1). A single pup in the mevalonic acid control group (Group 2) had no oral opening, no eyes and displaced ears; as a result, the incidence of this observation was significantly reduced (p50.01)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.1.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 (rng/dL) of PFOS alone Parameter I Cholesterol I Glucose I LDL-Dir I HDL-Dir Group 1 I 84.3 c 13.3 I 98.9 c 11.7 I 7.1 55.0 I 43.3 c 19.0 (Vehicle) (8) (8) (8) (8) Group 12 78.1 ? 10.5 104.9 c 7.8 10.8 c 4.4 43.5 c 11.2 1.6 mg/kg (8) (8) (8) (8) Group 13 72.3 c 6.3 96.3 rt 18.0 11.323.6 41.0k 11.7 II Triglycerides 313.5 c 233.6 (8) 194.3c65.2 (8) 201.0c87.5 II Mevalonic Acid 47.7 c 63.8 (8) 20.6 c 5.7 (8) 86.2 c 177.0 The total and free thyroid hormones levels for T3 and Tq in dams administered 1.6 or 2 mg/kg PFOS alone were significantly reduced @10.01), compared to the vehicle control group. 418-018:PAGE 111-10 Parameter Group 1 (Vehicle) Group 12 1.6 mglkg Grow 13 Total T4 ugldL 0.584 t 0.482 (8) 0.000 t O.OoO** (8) 0.000tO.OoO** Total T1 ngIdL 73.979 t 21.234 (8) 47.503 +10.268** (8) 42.440+8.271** Free T1 Pg/d 0.579 t0.209 (8) 0.140 t 0.117** (7) 0.157t0.161** TSH nglmL 1.815 t0.794 (8) 2.346 f 1.263 (7) 1.514t0.973 Free T4 ng1dL 0.299 t0.092 (8) 0.066 +0.038** (8) 0.055 tO.023** Liver cholesterol levels were significantly decreased (p10.05 or ~50.01i)n 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. Parameter I Cholesterol Group 1 3.0 -c 0.38 (Vehicle) (8) Group 12 2.6 t 0.22* 1.6 mgkg Group 13 (8) 2.4 rt 0.26** 2 mdkg (8) LDL-Dir 0.5 t 0.27 (8) 0.7 f 0.24 (8) 0.6 f0.12 (8) HDL-Dir 0.5 f0.12 (8) 0.6 tO.10 (8) 0.5 c 0.06 (8) Triglycerides 8.0 t 1.66 (8) 7.7 f 0.71 (8) 6.6 f 0.82 (8) The clinical chemistry parameter of low density lipoprotein (LDL) was significantly increased (p50.05 orp<O.Ol), albeit not dosage-dependent,in the DG 21 dams that were coadministered mevalonic acid and PFOS. I Clinical Chemistry Values (rng/dL) of PFOS plus Mevalonic Acid Parameter I Cholesterol 1 Glucose I LDL-Dir I HDL-Dir Triglycerides I Mevalonic Acid Group2 1 92.5k14.6 I 108.5t11.9 1 2.923.6 1 3 2 . 4 ~ 2 0 . 01453.9~307.8I 1663.8~415.6 (Mevalonic) Group3 (8) 91.0f21.4 (8) (8) 108.1 ~ 1 5 . 4 11.8-c9.0** (8) 35.1 f12.1 (8) 305.9f85.1 (8) 136932240.1 1.6 m g k g Group 4 (8) (8) 88.9 f 23.1 106.0 t 14.0 (8) 9.6 f 5.0* (8) (8) 47.8 f 16.2 243.6 c 270.8 (8) 1482.5 t 521.9 2 mglkg (8) (8) (8) (8) (8) (8) The total and free thyroid hormones levels for T3 and Tq in DG 21 dams coadministered mevalonic acid and 1.6 or 2 mgkg PFOS were significantlyreduced (p10.05 or p<O.Ol), compared to the vehicle control group. * Significantly different from control group;~ 5 0 . 0 5 . ** Significantly different from control group;~ 5 0 .I0. Parameter I (Mevalonic) I Total T4 ug/dL (8) Total T1 ng/dL (8) Free T3 pg/mL (7) 418-018:PAGE 111-11 TSH ng/mL (6) Free T4 ng1dL (8) Liver cholesterol and triglyceride levels were significantly reduced (p50.01) in dams coadministered mevalonic acid and 1.6 or 2 mgkg PFOS. Liver LDL and HDL levels were significantly increased in dams coadministeredmevalonic acid and 1.6 or 2 mgkg PFOS. Parameter Group 2 (Mevalonic) Group 3 1.6 rngkg Group 4 2 mgkg Cholesterol 3.0 -r- 0.34 (8) 2.5 t0.12"" (8) 2.4 2 0.30** (8) * LDL-Dir 0.5 0.24 (8) 0.8 f 0.08"" (8) 0.8 f 0.12"" (8) HDL-Dir 0.5 f 0.08 (8) 0.6 i 0.03** *(8) 0.6 0.05 (8) Triglycerides 9.8 f 1.45 (8) 7.5 t 0.55"" 7.0 *(8) 1.35"" (8) Clinical chemistry parameters were generally unaffected in the DG 21 dams that were treated with cholesterol and PFOS. 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<O.Ol), compared to the vehicle control group. * Significantly different from control group; ~50.05. ** Significantly different from control group; ~ 5 0 .10. 4 18-018:PAGE 111-12 Parameter Group 5 (Cholesterol) Group 6 1.6 mglkg Group 7 2 mdkg Total T4 ugldL 0.540 f 0.529 (8) 0.013 f 0.035** (8) 0.000 2 O.OOO** (8) Total T3 ng/dL 76.900 t 21.899 (8) 52.031 t 9.331** (8) 49.020 2 8.527** (8) Free T? pg/mL 0.694 t 0.256 (7) 0.235 f 0.123** (8) 0.246 f 0.180** (8) TSH ng/mL 2.544 t 0.759 (5) 1,4522 0.691 (6) 2.013 rt 0.854 (6) Free T4 ngldL 0.281 t 0.125 (7) 0.134 2 0.086"" (8) 0.1 10 2 0.082** (8) Liver triglyceride levels were significantlyreduced (p10.01) 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 coadministeredcholesterol and 1.6 or 2 mgkg PFOS. Liver cholesterol parameters were generally unaffected in the dams that were treated with 1.6 or 2 mg/kg PFOS plus Cholesterol. Parameter I (Cholesterol) Group 6 1.6 mgkg Group 7 2 mgkg Cholesterol (8) 2.7 2 0.29 (8) 2.7 -c 0.21 (8) I LDL-Dir I *(8) 0.8 0.23** (8) 0.8 t 0.18%" (8) HDL-Dir 0.6 *(8) 0.10 (8) 0.6 2 0.09 (8) Trig1ycerides (8) 7.7 f 0.72** (8) 7.5 .+ 0.70"" (8) PFOS concentrations were significantlyincreased (p10.05)in the sera of DG 21 dams administered 1.6 or 2 mgkg PFOS and in dams coadministeredCholesterol or Mevalonic acid and 1.6 or 2 mgkg PFOS. PFOS concentrations were significantly increased (p10.05)in the livers of dams administered 1.6 or 2 mgkg PFOS. ** Significantly different from control group;~ 5 0 .10 4 18-018:PAGE 111-13 Pararne ter Group 1 (Vehicle) Group 2 (Mevalonic) Group 3 1.6 rng/kg Group 4 2 mg/kg Group 5 (Cholesterol) Group 6 ~ 1.6 r n g k g Group 7 2 rng/kg Group 8 Group 9 Group 10 Seraa (ug/mL) 0.0 f 0.00 (6) 0.0 f 0.02 (6) 55.8 +. 8.88" (6) 82.4 f 22.55" (6) 0.0 +. 0.00" (6) 79.9 r 15.48" (6) 370.8 +. 214.36" (6) Livera (ppm) N.A. N.A. N.A. N.A. N.A. N.A. N.A. 1.2 mgkg Group 12 1.6 mg/kg Group 13 1Y.A. 142.2 f 24.24" (6) 124.7 +- 19.71" 1Y.N. N.A. KT A vv I N.A. = Not Analyzed a = Sponsor selected the subsets to be analyzed. K.1.b. DG 21 Fetuses The clinical chemistry parameters of cholesterol and low density lipoprotein (LDL) were significantly increased @l0.05orplO.O1) in the DG 21 fetuses from dams that were treated with 1.6 or 2 mg/kg PFOS alone. * Significantly different from control group; ~ 5 0 . 0 5 . 4 18-018:PAGE 111-14 Parameter Cholesterol Group1 (Vehicle) I 50.8-r-8.0 (8) Glucose 63.855.9 (8) LDL-Dir 27358.0 (8) HDL-Dir 13.021.9 (8) Triglycerides 34.4+5.3 (8) Mevalonic Acid 54.5+.40.5 (7) The level of free T4 was significantlyreduced (p10.01) in the DG 21 fetuses from dams that were treated with 1.6 or 2 mgkg PFOS alone. Parameter II TotalT,, ug/dL- I I Total T? ngIdL" I (Vehicle) I (7) (4) 1.6 m&g I (8) (8) Free T? PglmL (0) (1) TSH ng/& Free Ta ngldL (0) (1) (0) I (3) Liver HDL clinical chemistry parameters were significantly increased (p50.01) in DG 2 1 fetuses from dams administered 1.6 or 2 mgkg 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. Parameter (Vehicle) Group 12 1.6 mg/kg Grouu 13 I Cholesterol I LDL-Dir *(8) 3.4 0.38 (8) 3.2 -r- 0.34 (8) 0 . 0 0 0 ~0.0000 0.000 *(8) 0.0000 HDL-Dir 1 (8) 0.02 +. 0.006" (8) 0.02 +. 0.015" Trig1ycerides *(8) 5.9 0.93 (8) 5.6 c 0.91 The clinical chemistry parameters of cholesterol and low density lipoprotein (LDL) were significantly increased (pSO.01) and triglycerides were significantly decreased (pSO.01) in the DG 2 1 fetuses from dams that were treated with mevalonic acid and 1.6 and/or 2 mg/kg PFOS. * Significantly different from control group;~ 1 0 . 0 5 . ** Significantly different from control group; ~ 5 0 .10. 4 18-018:PAGE 111-15 Clinical C p3GziF Cholesterol Glucose (Mevalonic) 1.6 mg/kg 53.1 c 5.5 63.4 f 15.0 ~ 68.3 * 13.4%" 69.6 c 6.2 ~ Group 4 ~ 63.1 c 11.7 ~ 74.5 f 12.3 LDL-Dir HDL-Dir Triglycerides Mevalonic Acid 29.4 c 5.3 13.9 ~f1:.2 36.4 c 8.7 18971.4c (8) (8) (8) 5681.5 48.3 c 8.1** 16.1 c3.0 31.8c7.1 (7) 16562.5 c (8) (8) (8) 7 160.4 46.9 t9.4"" I 15.5 2 1.9 1 25.1 +8.1** I (8) 20175.0 f The level of free Tq was significantly reduced (p10.01) in the DG 21 fetuses from dams Parameter Group 2 (Mevalonic) Group 3 1.6 mgkg Group 4 2 mdkg Total T4 *ug/dL 0.000 0.000 (7) 0.000 c 0.000 (8) 0.000 ~f0:.000 (8) Total T3 ng/dL 2.810 f 4.867 *( 3 ) 1.096 1.379 *(7) 2.928 2.747 (6) Free T3 pg/d (0) 0.000 f 0.000 (1) 0.000 c 0.000 (2) TSH ng/mL (0) (0) (0) Free T4 ng/dL 0.070 c 0.000 (1) 0.000 c o.ooo** *(2) 0.000 0.000"" (4) Liver cholesterol, HDL and triglyceride levels were significantly increased (p20.05 or p10.01) in the DG 21 fetuses of dams that were treated with 1.6 or 2 m g k g PFOS plus Mevalonic Acid. Parameter Group 2 (Mevalonic) Group 3 1.6 mg/kg Group 4 2 mg/kg Cholesterol 3.6 c 0.49 (8) 4.5 2 0.48** (8) 5.1 f 0.82** (8) LDL-Dir o.ooo* 0.0000 (8) 0.000~0t .0000 (8) 0.000 f 0.0000 (8) HDL-Dir 0.01 cO.009 (8) 0.04 c 0.019** (8) 0.03 0.018" (8) * Trig1ycerides 5.4 1.25 (8) 8.0 -t 1.16** 9.6 *(8) 2.39"" (8) The clinical chemistry parameters of cholesterol, low density lipoprotein (LDL) and high density lipoprotein (HDL) were significantly increased (p10.05 or p10.01) in the DG 21 fetuses from dams that were treated with cholesterol and 1.6or 2 mgkg PFOS. ** Significantly different from control group; pFO.01. (Cholesterol) I (8) (8) (8) 4 18-018:PAGE 111-16 (8) (8) The level of free Tq was significantlyreduced Cp<O.Ol) in the DG 21 fetuses from dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS. Parameter Group 5 (Cholesterol) I Group 6 1.6mgkg Total T4 ug/dL 0.000k0.000 (8) 0.000+.0.000 (7) * Total T3 ng/dL 1.143+.1.306 (6) 0.760k1.389 (5) Free T3 PdmL O.OOOkO.000 (2) 0.00Ok0.000 (2) TSH ng/mL 0.000+0.000 (0) 0.040r0.000 (1) Free T4 ng/dL 0.037r 0.015 (3) 0.000+0.000** (4) Liver cholesterol and triglyceride levels were increased or significantlyincreased Cp10.01) in the DG 2 1 fetuses from dams coadministered cholesterol and 1.6 or 2 mgkg 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. Parameter Group 5 1 Cholesterol I LDL-Dir I 3.6 50.29 I 0.00Ok 0.0000 I (Cholesterol) (8) (8) Group 6 4.4 k 0.40"" 0.000k 0.0000 1.6 mgkg (8) (8) Grouo 7 4.0 20.68 0.000 k 0.0000 HDL-Dir 0.02 k 0.01 9 (8) 0.03 +. 0.010 (8) 0.04 5 0.023 Triglycerides 5.5 f 0.73 (8) 7.8 k 1.23"" (8) 7.1 r 1.15"" PFOS concentrations were significantly increased (p10.01) 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 I .6 or 2 mgkg PFOS. PFOS concentrations were significantly increased Cp10.05) in the livers of fetuses administered 1.6 or 2 mg/kg PFOS. * Significantly different from control group; ~ 5 0 . 0 5 . ** Significantly different from control group; ~10.01. 4 18-018:PAGE 111-17 Parameter Group 1 (Vehicle) Group 2 (Mevalonic) Group 3 1.6 mgkg Group 4 2 mglkg Group 5 (Cholesterol) Group 6 1.6 mglkg Group 7 2 mglkg Group 8 0.4 mglkg Group 9 0.8 mgkg Group 10 1 mglkg Group 11 1.2 mglkg Group 12 1.6 mgkg Group 13 2 mskg (ug1mL) 0.0 t 0.00 (6) 0.1 +. 0.08 (6) 207.2 +. 25.76** (6) 251.O t 55.86** (6) 0.1 k 0.03 (6) 154.5 s 32.43** (6) 165.7 t 50.13** (6) N.A. N.A. N.A. N.A. 141.8 * 33.10** (6) 169.8 s 37.91** (6) (ppm) N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. K.2. Lactation Day 5 K.2.a. Dams The clinical chemistry parameter of serum cholesterol was significantly decreased (p10.01) 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 (p10.01) in lactating dams treated with 2 mgkg PFOS alone and the clinical chemistry parameter of triglycerides was significantlydecreased (p10.05or ~ 1 0 . 0 1i)n lactating dams treated with 1.6 or 2 mg/kg PFOS alone, compared to the vehicle control group. * Significantly different from control group;~ 5 0 . 0 5 . ** Significantly different from control group; ~ 1 0 . 0 1 . 4 18-018:PAGE 111-18 Parameter Group 1 (Vehicle) Group8 0.4 mg/kg Group9 0.8 mg/kg Group 10 1 mg/kg Group11 1.2 m a g Group 12 1.6 mg/kg Group 13 2 m@g Cholesterol [Blood] 77.6k15.2 (19) 65.0+13.2** (20) 59.1t13.3** (17) 58.5 t 11.8"" (19) 59.9211.7** (18) Cholesterol [Milk] 96.7k53.9 (19) 71.1 k62.8 (20) 68.2t52.6 (17) 59.8 t 51.2 (18) 46.8k41.5 (18) 56.6 t 14.0"" (17) 59.6 t 10.2%" (19) 64.6 k 30.4 (1 3 ) 74.0 k 41.7 (5) Glucose 166.1 k28.4 (19) 169.6k26.7 (20) 163.0k17.3 (17) 171.9 k 19.9 (19) 172.5k17.7 (18) 178.7 +. 19.0 (17) 189.2 k 32.2"" (19) LDL-Dir 4.0t3.0 (19) 3.2223 (20) 3.222.1 (17) 3.7 k 1.9 (19) 2.8k2.2 (18) 3.6 t 2.6 (17) 4.6 k 1.8 (19) HDL-Dir 58.2k20.1 (19) 51.2k17.2 (20) 47.5t14.6 (17) 49.0 k 11.0 (19) 48.9216.2 (18) 49.5 k 10.3 (17) 53.1 t 9.4 (19) Triglyceride S 152.4297.6 (19) 130.1 278.3 (20) 127.7k66.5 (17) 101.2 k 52.7 (19) 141.1k74.3 (18) 92.2 k 58.9" (17) 85.5 k 35.0"" (19) Mevalonic Acid 31.0k19.8 (18) Not Evaluated Not Evaluated Not Evaluated 83.12 178.8"" (18) 32.2 k 20.9 (16) 29.6 t 14.4 (15) The total and free T4 levels in lactating dams administered 0.4 (free T4 only), 0.8, 1, 1.2, 1.6 or 2 mgkg PFOS were significantly reduced (p10.05or p10.01j and total and free T3 levels in lactating dams administered 1.2, 1.6 or 2 mg/kg PFOS were significantly reduced ($10.05or p10.01j, compared to the vehicle control group. Parameter Group 1 (Vehicle) Group 8 0.4 mg/kg Group 9 0.8 mg/kg Group 10 1m a g Group 11 1.2 m g k g Group 12 1.6 mg/kg Group 13 2 m@g Total T4 ug/dL 1.454 k 0.660 (19) 0.808 k 0.414 (10) 0.596 s 0.443" (8) 0.728 20.243"" (9) 0.283 k 0.320"" (18) 0.272 k 0.172"" (17) 0.235 k 0.147** (19) Total T3 ng/dL 74.674 t 18.977 (19) 72.912 k 13.470 (10) 63.780 k 6.684 (8) 62.348 rt 13.180 (9) 52.869 t 14.974"" (18) 47.032 k 19.971"" (17) 53.275 f 17.270"" (19) Free T1 Pg/d 0.638 k 0.297 (19) 0.698 t 0.246 (10) 0.588 k 0.152 (8) 0.552 t 0.230 (9) 0.41 1 k 0.265" (18) 0.256 t 0.230** (17) 0.3.53 t 0.236"" (18) TSH ng/mL 1.633 k 0.992 (1 8) 1.142 k 0.226 (9) 1.437 t0.918 (7) 1.1 10 k 0.523 (8) 1.447 k 1.029 (17 ) 1.665 k 0.768 (1 7) 1.533 k 0.681 (1 8) Free T4 ng/dL 0.788 k 0.219 (19) 0.462 k 0.1 14"" (10) 0.279 t 0.060"" (8) 0.277 k 0.095"" (9) 0.260 k 0.122%" (18) 0.253 k 0.067"" (17) 0.259 k 0.078"" (18) Liver triglyceride levels were significantly increased (p10.01) 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. * Significantlydifferent from control group; ~ 5 0 . 0 5 . ** Significantlydifferent from control group;p50.01. 4 18-018:PAGE111-19 Parameter Group 1 (Vehicle) Group 8 0.4 mg/kg Group 9 0.8 mgkg Group 10 1 mg/kg Group 11 1.2 mgkg Group 12 1.6 mg/kg Group 13 2m a g I I Cholesterol I 2.4 k 1.29 I (19) 2.6 2 0.67 (20) 2.4 2 0.59 (17) ~ 2.5 20.54 (19) 2.5 2 0.83 (18) 2.6 k 0.79 (17) 2.9 k 1.08 (19) LDL-Dir 0.00 rt 0.014 (19) 0.01 2 0.034 (20) 0.05 -t 0.093 (17) 0.00 -t 0.004 (19) 0.05 2 0.106 (18) 0.01 2 0.038 (17) 0.03 2 0.054 (19) HDL-Dir 0.13 k 0.094 (19) 0.22-to.115 (20) 0.23 -t 0.130 (17) 0.20 2 0.093 (19) 0.26 k 0.135 (18) 0.21 20.104 (17) 0.22 k 0.1 17 (19) Triglycerides 7.7 2 2.90 (19) 8.4 2 1.70 (20) 8.6 2 2.47 (17) 11.8 2 12.02 (19) 9.7 2 3.44 (18) 13.4 k 7.20"" (17) 16.0 f 5.38** (19) The clinical chemistry parameters of serum cholesterol was significantly decreased (p10.01) in the lactating dams that were treated with mevalonic acid and 1.6 or 2 mgkg PFOS and cholesterol in the milk was significantlyincreased (p10.05i)n the lactating dams that were treated with mevalonic acid and 2 mgkg PFOS. Parameter Group 2 (Mevalonic) Group3 1.6 mg/kg Group4 2 mgkg Cholesterol [Blood] 80.5 k 15.6 (1 8) 63.6+14.6** (18) 67.3-t11.2** (19) Cholesterol [Milk] 81.4252.4 (17) 70.5k46.2 (12) 188.0i:114.6" (2) Glucose 174.3k39.2 (18) 174.9k24.1 (18) 187.6k30.9 (19) LDL-Dir 3.722.9 (18) 3.4k2.0 (18) 3.7k3.1 (19) HDL-Dir 63.7t18.1 (18) 54.2212.1 (18) 55.0k10.1 (19) Triglycerides 135.6k58.6 (18) 103.3243.5 (18) 118.7k50.7 (19) Mevalonic Acid 201.4k 135.9 (18) 1123.0k 2000.2" (1 8) 18662.4-t 70304.8" (19) The levels of total and free T4 were significantly reduced (p50.01) in the lactating dams that were treated with mevalonic acid and 1.6 or 2 mgkg PFOS. Parameter Group 2 (Mevalonic) I Grouu 3 1.6 mg/kg Total Ta ug/dL 2.608 k 1.083" (5) 0.182 f 0.243"" (4) Total T? ng/dL88.864 f 19.532 (5) 65.752 k 6.320 (4) Free T? Pg/d0.822 2 0.295 (5) 0.695 k0.131 (4) TSH ng/mL 1.962 k 1.655 (5) 1.228 k 1.076 (4) Free Td ng/dL 0.894 2 0.248 (5) 0.270 k 0.067"" (4) Liver triglyceride levels were significantly increased (piO.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;~ 5 0 . 0 5 . ** Significantly different from control group;~ 5 0 . 10. mg/kg PFOS plus Mevalonic Acid. 418-018:PAGE 111-20 Parameter GrouD 2 (Mevalonic) Group 3 1.6 mg/kg Group 4 2m g k Cholesterol 2.3 c 1.00 (18) 2.2 c 0.65 (18) 2.7 c 1.29 . (19) LDL-Dir 0.01 rt0.041 (18) 0.02 c 0.064 (18) 0.01 c 0.031 (19) HDL-Dir 0.16 c0.122 (18) 0.19 c 0.134 (18) 0.18 c0.129 (19) Trig1ycerides 7.5 c 2.05 (18) 10.3 c 6.24 (1 8) 15.2 rt 5.49** (19) Levels of cholesterol in the blood and triglycerides were significantly decreased @<O.O 1) in lactating dams that were treated with cholesterol and 1.6 or 2 mg/kg PFOS and the blood level of glucose was significantlyincreased @<0.05)in lactating dams that were treated with cholesterol and 2 mg/kg PFOS. Parameter Group 5 (Cholesterol) Group 6 1.6 mg/kg Group 7 2 mg/kg Cholesterol [Blood] 75.9 +. 17.5 (17) 56.2 t 12.2** (20) 58.0 +. 15.2** (19) Cholesterol [Milk] 80.2 +. 57.9 (17) 81.9 c 52.5 (12) 57.8 c 3.8 (4) Glucose 168.8 t 29.2 (17) 170.2 c 20.8 (20) 190.6 c 28.5* (19) LDL-Dir 3.8 c 3.0 (17) 4.4 c 2.8 (20) 4.9 c 2.4 (19) HDL-Dir 55.6 c 19.1 (17) 48.6c 11.1 (20) 50.9 c 13.3 (19) Triglycerides 146.1 c 64.8 (17) 87.2+.32.1** (20) 85.0 t 36.1** (19) The level of total and free T3 and T4 were significantly reduced (p<0.05 or ~ 1 0 . 0 1i)n the lactating dams that were treated with cholesterol and 1.6 or 2 mgkg PFOS. Liver triglyceride levels were significantly increased (p10.05) in lactating dams coadministeredcholesterol and 2 mgkg PFOS. All other liver clinical chemistry parameters were generally unaffected in the DL 5 dams that were treated with 1.6 or 2 mgkg PFOS plus Mevalonic Acid. * Significantly different from control group; ~ 5 0 . 0 5 . ** Significantly different from control group;~ 5 0 . 0 1 . Parameter Group 5 (Cholesterol) Grouv 6 1.6 r n h g Group 7 Cholesterol 2.1 c 0.70 (17) 2.3 i:0.69 (20) 2.3 i:0.61 LDL-Dir 0.00 c 0.005 (17) 0.01 i:0.036 *(20) 0.00 0.000 4 18-018:PAGE 111-21 * HDL-Dir 0.16 0.096 (17) 0.20 c 0.1 15 (20) 0.19 k 0.078 Trig1yceridcs 7.6 c 1.68 (17) 10.1 k4.68 (20) 11.O i:4.83" Sera' Parameter Group I (ug/mL) 0.0 s 0.01 (Vehicle) Group 2 (6) 12.7 k 27.37 (Mevalonic) Group 3 (6) 127.0 s 24.41" 1.6 mgkg Group 4 (6) 189.2 t 48.20" 2 mglkg Group 5 (6) 0.7 t 1.62 (Cholesterol) Group 6 (6) 104.3 t 14.57" 1.6 mgkg Group 7 (6) 144.8 t 7.60" 2 mglkg Group 8 0.4 mgkg (6) 27.2 t 18.72" I (6) 0.8 mgkg Group 10 1 mglkg Group 11 1.2 mglkg Group 12 1.6 mglkg Group 13 2 mgkg (6) 52.3 _t 26.02" (6) 86.0 t 10.22" (6) 169.0 s 32.12" (6) 134.0 t 26.60" (6) Liver' (ppm) 0.5 s 0.87 (6) hT A 1Y.A. N.A. N.A. N.A. N.A. N.A. 47.9 t 5.15* (4) 1Y.A. N.A. N.A. 110.2 t 13.43" (5) 145.8 s 20.18" (6) * Significantly different from control group; ~50.05. 418-018:PAGE 111-22 K.2.b. LD 5 F l Generation Pups The clinical chemistry parameters measured were not significantlydifferent in the F1 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. Parameter Group I (Vehicle) Group8 0.4 mg/kg Group 9 0.8 mg/kg Group 10 1 mgkg Group 11 I .2 m@g Group 12 , 1.6 mg/kg Group 13 2 mg/kg Cholesterol 112.5 t 10.6 (12) 111.9+11.3 (15) 114.8 2 11.1 (13) 113.1 rt 16.8 (14) 124.7 t 23.4 (14) 132.3 2 33.9 (6) 128.0 t 38.3 (3) Glucose 255.8 t 61.8 (12) 244.5260.1 (15) 220.3 rt 56.2 (13) 210.0 2 47.0 (14) 217.2 t 60.6 (14) 222.0 2 20.8 (6) 229.3 2 17.2 (3) LDL-Dir 33.5 t 8.7 (12) 29.1 27.4 (15) 31.5 27.6 (13) 29.4 2 5.3 (14) 34.8 2 15.0 (14) 40.3 2 20.1 (6) 44.7 2 31.6 (3) HDL-Dir 29.8 2 7.9 (12) 32.7 rt 6.9 (15) 38.1 t 11.0 (13 ) 36.3 2 6.6 (14) 31.9 rt5.8 (14) 36.0 2 7.0 (6) 34.3 t 4.0 (3) Triglycerides 214.2 rt 51.8 (12) 234.9 rt 95.0 (15) 179.2 2 66.5 (13) 197.8 k 64.4 (14) 230.3 2 78.5 (14) 231.8 k52.7 (6) 213.0 t 56.3 (3) Mevalonic Acid 22.6 t5.3 (9) Not Evaluated Not Evaluated Not Evaluated 18.7 rt 4.0 (8) 20.6 2 4.0 (4) (0) The free and total Tq 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<O.Ol), compared to the vehicle control group. The thyroid stimulating hormone (TSH) was significantly increased (p50.05 or ~50.01a)t 1 and 1.6 mg/kg PFOS, but not in a dosage-dependentfashion. Parameter Group 1 (Vehicle) Group 8 0.4 mg/kg 0.8 m i k g 1 mg/kg Group 11 1.2 m a g G r o w 12 1.6 m3kg Group 13 2 mg/k Total T4 ug/dL 0.543 2 0.221 (12) I 0.000 2 O.OOO** (10) I (8) (9) 0.005 rtO.O14** 0.005 *(13) 0.012** (6) 0.000 k O.OOO** (3) Total T3 ng/dL 54.383 2 17.965 (12) 55.777 t 18.934 (9) (8) (7) 44.715 t 21.746 (11) 32.708 t 7.941 (6) 32.777 2 12.162 (3) Free T, pg/mL 0.039 2 0.086 (9) 0.016 t 0.036 (5) (5) (4) 0.003 t0.009 (10) 0.004 2 0.009 (5) 0.050 2 0.087 (3) TSH nglmL 1.018 20.166 (8) (0) (0) (1) 1.0102 0.246 (5) 1.450 2 0.339* (5) 1.500 t 0.000 (1) Free T4 ng/dL 0.090 t 0.024 (1 1) 0.018 t 0.009** (8) (7) (7) 0.011 t 0.009** (1 1) 0.010 2 0.006** (6) 0.010 2 0.010** (3) Liver triglyceride and LDL levels were decreased or significantly decreased (p50.05 or ~ 1 0 . 0 1i)n 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;~50.05. ** Significantly different from control group;~ 5 0 . 0 1 . 418-018:PAGE 111-23 Parameter Group 1 (Vehicle) Group 8 0.4 mgkg Group 9 0.8 mg/kg 1 mgkg Group 11 1.2 m a k g Grow 12 1.6 mgkg Group 13 Cholesterol 3.8 rt 0.66 (18) 3.9 rt 0.48 (20) 3.8 rt 0.30 (17) (18) 3.8 rt 0.62 (17) 3.5 rt 0.32 (10) 3.8 rt 0.62 LDL-Dir 0.012 k 0.0264 (18) 0.000 f 0.0022** (20) 0.000 f 0.0000*" (17) (18) I 0.000 rt0.0000** I (17) 0.003 rt 0.0095 (10) 0.000 rt 0.0000 HDL-Dir 0.10 rt 0.025 (18) 0.09 rt 0.03 1 (20) 0.09 rt 0.038 (17) (18) 0.09 rt 0.037 (17) 0.11 rt 0.032 (10) 0.09 rt 0.017 Trigl ycerides 18.1 rt7.94 (18) 14.0 rt 6.18 (20) 14.0 4.67 (17) (18) 12.0 f 6.85** (17) 11.4 rt 3.1 1 * (10) 10.8 rt 2.04 Liver triglyceride levels were decreased or significantly decreased Cp<0.05 or ~ 2 0 .10) in DL 5 male pups from lactating dams administered 0.4,0.8, 1, 1.2, 1.6 or 2 mgkg PFOS. All other liver clinical chemistry parameters were generally unaffected in the male pups whose dams were treated with 0.4 to 2 mgkg PFOS alone. Male Pups Liver Clinical Chemistry Values (mg/gm) of PFOS alone Parameter 1 Cholesterol LDL-Dir HDL-Dir Grouo 1 3.5 rt 0.49 0.003 f 0.0118 0.08 f 0.019 (Vehicle) Group 8 0.4 mg/kg Group 9 0.8 mg/kg Group 10 (18) 3.9 f 0.83 (20) 3.6 0.56 (17) 3.7 f 0.76 (18) 0.000 f 0.0000 (20) 0.000 rt 0.0000 (17) 0.003 f 0.0097 (18) 0.08 rt 0.043 (20) 0.08 f 0.041 (17) 0.10 f0.030 1m a g Group 1I 1.2 m a g Group 12 (18) 3.6 0.55 (18) 3.3 rt 0.52 (18) 0.000 f 0.0000 (18) 0.003 -c 0.0067 (18) 0.10 rt 0.042 (18) 0.09 rt 0.030 1.6 mg/kg Group 13 (10) 3.4 f 0.26 (10) 0.005 f 0.0100 (10) 0.09 f 0.033 2 mg/k (4) (4) (4) Trigl ycerides 17.3 f 8.05 (18) 15.4 f 6.76 (20) 14.4 f 6.46 (17) 11.1 f 3 . 2 1 * * (18) 10.9 f 4.80** (18) 11.4 f 7.55" (10) 7.4 f 1.54* (4) Levels of cholesterol, low density lipoproteins (LDL) and triglycerides in the blood were significantly increased Cp10.05 or ~10.01i)n DL 5 pups from lactating dams that were treated with mevalonic acid and 1.6 mgkg PFOS. * Significantly different from control group; ~50.05. ** Significantly different from control group; ~ 5 0 . 0 1 . 418-018:PAGE 111-24 Clinical Chemistry Values (mg/dL) of PFOS plus Mevalonic Acid Parameter I Cholesterol I Glucose I LDL-Dir I HDL-Dir I Triglycerides I Mevalonic Acid Group2 I 115.4k7.6 I 241.4288.4 I 35.3 k7.5 1 35.429.0 I 193.2269.9 I 335.3 k 229.2 (Mevalonic) (12) (12) (12) (12) (12) (1 1) Group 3 157.0 f23.7:g* 273.3 -c 65.1 50.8 f20.0* 33.0 k4.9 302.3 k 62.7** 270.8 k 85.8 1.6 mg/kg (7) (7) (7) (7) (7) (7) Group 4 0.00 k 0.000 0.00 k 0.000 0.00 2 0.000 0.00 k 0.000 0.00 k 0.000 195.2 k 203.4 2 mg/kg (0) (0) (0) (0) (0) (2) Levels of total Td were significantly decreased @<O.Ol) in DL 5 pups from lactating dams that were treated with mevalonic acid and 1.6 mg/kg PFOS. Parameter Group 2 (Mevalonic) Group 3 1.6 mg/kg Total T4 ugIdL 0.486 2 0.227 (5) 0.000 k O.OOO** (3) Total T3 ng/dL 83.420 k 31.720' (5) 70.633 k 24.823 (3) Free T3 Pg/d 0.497 k 0.490 (3) 0.00 k 0.000 (0) TSH nglmL 0.430 k 0.212 (2) 0.680 f 0.000 (1) Free T4 ng/dL 0.090 k 0.056 (5) 0.005 k 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 mgkg PFOS plus Mevalonic Acid. Parameter Group 2 (Mevalonic) Grouu 3 1.6 m a g Group 4 2 mdkp Cholesterol 3.8 k 0.64 (18) 3.3 k 0.86 (9) LDL-Dir 0.008 k 0.021 8 (18) 0.002 f0.0067 (9) HDL-Dir 0.11 k 0.039 (18) 0.08 k 0.030 (9) NO VALUES FOR THIS GROUP Trig1ycerides 18.4 k 8.93 (18) 14.1 f 6.00 (9) 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 mgkg PFOS plus Mevalonic Acid. * Significantly different from control group; ~ 5 0 . 0 5 . ** Significantly different from control group; ~50.01. a. Group 2 significantly different from Group 1;~ 5 0 . 10. 418-018:PAGE 111-25 Parameter Group 2 (Mevalonic) I Group 3 1.6 mg/kg Group 4 2 mgkg Cholesterol 3.6 f 0.69 *(18) 3.3 0.68 (10) LDL- Di r HDL-Dir 0.003 f 0.0096 0.09 f 0.026 (18) 0.008 f 0.02.53 *(18) 0.09 0.037 (10) (10) NO VALUES FOR THIS GROUP Triglycerides 1 3 . 0 7~.38 (18) 13.6 f 5.38 (10) Levels of low (LDL) and high (HDL) density lipoproteins were significantly increased (p10.05or p10.01) in the pups from lactating dams that were treated with cholesterol and 1.6 or 2 mgkg PFOS and the blood level of triglycerides was significantlydecreased (p10.05) in the pups from dams that were treated with cholesterol and 2 mgkg PFOS. Parameter Group 5 (Cholesterol) Group 6 1.6 mg/kg Group 7 2 mgkz * Cholesterol 117.8 9.1 (12) 130.0 +. 18.5 (8) 134.8 f 26.1 (5) Glucose 263.8 2 56.4 (12) 247.8 c 48.9 (8) 223.2 f 20.2 (5) LDL-Dir 26.8 f 12.2 (12) 41.0 k 11.4* (8) 55.2 f.22.5** (5) HDL-Dir 28.7 f 7.7 *(12) 39.2 3.6** 47.2 *(8) 12.8** (5) Triglycerides 321.3 f 208.8 (12) 232.9 f 74.4 (8) 120.4 f 36.6* (5) Levels of total T3 and free T4 were significantly decreased (p10.05 or p10.01) in pups from lactating dams that were treated with cholesterol and 1.6 and/or 2 mgkg PFOS. Liver clinical chemistry parameters were generally unaffected in the DL 5 female pups of lactating dams that were treated with 1.6 or 2 mgkg PFOS plus Mevalonic Acid. Parameter Group 5 (Cholesterol) Group 6 1.6 mg/kg Group 7 2 mg/kg Cholesterol 3.8 0.56 (17) 3.4 f 0.54 (1 1) 4.0 f 0.14 (4) LDL-Dir 0.01 1 f 0.027 1 (17) 0.000 f 0.0000 (11) 0.000 f 0.0000 (4) HDL-Dir 0.09 2 0.040 (17) 0.08 f 0.03 1 (1 1) 0.10 f 0.037 (4) Trig1ycerides 17.9 2 7.29 *(17) 12.9 4.36 (1 1 ) 13.5 *5.16 (4) * Significantly different from control group; pF0.05. ** Significantly different from control group;~ 5 0 .10. a. Group 5 significantly different from Group 1;pF0.01. 418-018:PAGE 111-26 Liver triglyceride levels were significantly reduced @50.05or ~50.01i)n 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. Parameter Group 5 (Cholesterol) Grow 6 1.6 mgkg Grow 7 Cholesterol 3.7 f 0.45 (17) 3.4 c 0.45 (8) 4.1 c 0.73 LDL-Dir 0.004 c 0.0100 (17) 0.000 c 0.0000 (8) 0.000 f 0.0000 HDL-Dir 0.08 c 0.029 (17) 0.10 c 0.031 (8) 0.08 c 0.035 Trig1ycerides 18.3 c 6.80 (17) 10.5 c 3.95"" (8) 10.6 24.12" Seraa Parameter Group 1 (ug/mL) 0.0 t 0.02 (Vehicle) Group 2 (6) 2.8 2 3.12 (Mevalonic) Group 3 (6) 123.7 f 18.58* 1.6 mglkg (3) Group 4 N.A. 2 mglkg Group 5 0.2 2 0.19 (Cholesterol) Group 6 (6) 108.5 f 7.78 1.6 mglkg Group 7 (2) 155.0 t 0.00 2 mglkg Group 8 (1) 36.2 t 3.84 , 0.4 m&g Group 9 (6) 53.1 t 27.67 I I I 0.8 mgkg Group 10 1 mglkg (6) 84.5 t 17.52 (6) Livera (PPm) BQL (6) N.A. N.A. N.A. N.A. N.A. N.A. 73.4 t 30.83 (6) N.A. hiYT.n A . V " I Group 12 XT A " V I Group 13 138.0 k 0.00 2 makg (1) 274.0 2 127.80** (5) 245.2 k 93.36** ( 4) * Significantly different from control group; ~ 1 0 . 0 5 . ** Significantly different from control group; ~ 1 0 . 0 1 . 418-018:PAGE 111-27 L. Histopathology of Hearts and Thyroids of F1 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 1 1059 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. 418-018:PAGE 111-28 REFERENCES 1. Study Design as Modification of U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Federal Register, September 22, 1994, Vol. 59, No. 183. 2. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. 3. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standardfor Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 4. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionh-ecommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 3 15; 28 October): 1-17. 5. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield,VA 22161. 6. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 7. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Cr1:CDBBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research.) 8. Institute of Laboratory Animal Resources (1996). Guidefor the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 9. Salewski, E. (1964). Fiirbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. 10. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 240-241. 11. Sokal, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. 12. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 418-018:PAGE 111-29 13. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-1121. 14. Sokal, R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 15. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):24 1-252. 16. Siegel, S. (1956). Nonparametric Statistics for the Behavioral Sciences, McGrawHill, New York, pp. 96-104. APPENDIX A REPORT FIGURES 418-018:PAGE A-1 0 In 0 In 0 In 0 In 0 v) 0 Iun. N d 0 0 mi. Imn N m 0 m i. In N N N N 0 N (3)l H 3 1 3 M PROTOCOL 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 450 I I Fo GENERATION FEMALE RATS Figure 2 - Groups 2,3,4 425 1 400 375 350 h c3 v 1 I- I 325 I (3 ! - 300 1- J GROUP 3 GROUP 4 ! 1 *p<0.05 (from group 2) **p<O.Ol (from group 2) 275 soP c. 250 0 c. PP 225 70m I ' b 4 4 h 8 b ' 4 k 200 'I 1 8 I!? d2 d9 3644a DAY OF STUDY 3! k 6 'I 1'0 1'1 1b 113 I$I!? 1's 1'7 I 8 16 20 21 DAY OF GESTATION DAY OF LACTATION a. Last value recorded before cohabitaton PROTOCOL 418-018: ONE GENERATION REPRODUCTIONSTUDY OF PFOS - MEVALONICAClDlCHOLESTEROL CHALLENGEAND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) BODY WEIGHTS 450 ~ Fo GENERATION FEMALE RATS Figure 3 - Groups 5,6,7 I , I I I 425 GROUP 5 400 375 350 3 lI: 325 cr, 300 Ll GROUP 6 0 GROUP 7 1 , #p<0.05 *p<0.05 (from (from group group I) 5) I 275 P c3. b 250 rn CL 225 I 6 200 1 , 1 15 212 29 d642a DAY OF STUDY 6 4 !! 3 b 5 6 7! 8 9 10 1'1 I 12 l 13 l 14 1 15 16 1'7 Ib lb d0 d1 ' I I I I 1 A DAY OF GESTATION DAY OF LACTATION a. Last value recorded before cohabitaton. APPENDIX B REPORT TABLES PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 1 (PAGE I): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 8 10 11 12 13 DOSAGE MG/KG PFOS VEHICLE CONTROL 0.4 1 1.2 1.6 2 --.----._-.-._-_.__....---.--------.---------.---.---..----.-----. -PR_E_C_OH~A_BI_T_AT- I~ON _(__D_AY 1 OF STU_D_Y_T_O- T~ HE DAY___O_F___C_ O~ HABITATI~ ON): - MAXIMUM POSSIBLE INCIDENCE 1176/ 28 840/ 20 20 MORTALITY 0 0 LOCALIZED ALOPECIA: TOTAL 611 4 29/ 2 0 UNDERSIDE 14/ 2 o/ 0 0 LIMBS 47/ 2 29/ 2 0 lNCISORS: TOTAL MISSING/BROKEN MISALIGNED o/ 0 5/ 1 2 o/ 0 o/ 0 1 o/ 0 5/ 1 1 CHROMODACRYORRHEA o/ 0 8/ 1 1 FOREPAWS AND LEFT FORELIMB: SCABS o/ 0 25/ 1 0 TAIL BENT 40/ 1 o/ 0 0 RIGHT FOREPAW: 2ND DIGIT MISSING 16/ 1 o/ 0 0 RIGHT FOREPAW: 2ND DIGIT SWOL,LEN 13/ 1 o/ 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. PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 1 (PAGE 2): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 8 DOSAGE MG/KG PFOS VEHICLE CONTROL 0 . 4 _ _ _ _ _ - - _ _ _ _ _ _ _ _ _ _ _ ___ __ _ _ __..._ . .-_ . .....- _-_-_ -_ - _ - - _ _ _ _ PRESUMED GESTATION: 12 1.6 MAXIMUM POSSIBLE INCIDENCE 615/ 28 434/ 20 594/ 27 MORTALITY 0 LOCALIZED ALOPECIA: TOTAL LIMBS NECK UNDERSIDE BACK 80/ 5 43/ 2 16/ 2 27/ 2 o/ 0 INCISORS: TOTAL MISSING/BROKEN MISALIGNED SWOLLEN SNOUT 7/ 2 7/ 2 o/ 0 o/ 0 CHROMODACRYORRHEA CHROMORHINORRHEA TAIL BENT RIGHT FOREPAW: 2ND DIGIT MISSING RED PERIVAGINAL SUBSTANCE 22/ 1 22/ 1 I/ 1 o/ 0 o/ 0 o/ 0 o/ 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 (~50.01). 13 2 610,' 2 8 o/ 0 o/ 0 o/ 0 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 1 (PAGE 3): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS -LACTATIO~ N:- MAXIMUM POSSIBLE INCIDENCE LOCALIZED ALOPECIA: TOTAL LIMBS NECK UNDERSIDE BACK INCISORS: TOTAL MISSING/BROKEN MISALIGNED 1 8 VEHICLE CONTROL 0.4 95/ 19 15/ 3 5/ 1 o/ 0 10/ 2 o/ 0 l o o / 20 10/ 2 10/ 2 o/ 0 o/ 0 o/ 0 9 0.8 8 5 / 17 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 10 1 93/ 19 17/ 4 13/ 3 o/ 0 4/ 1 o/ 0 11 1.2 90/ 18 13/ 3 9/ 2 o/ 0 4/ 1 8/ 2 12 1.6 76/ 17 15/ 3 10/ 2 I/ 1 5/ 1 o/ 0 13 2 61/ 19 9/ 3 6/ 3 5/ 1 o/ 0 o/ 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. bc-l PROTOCOL 418-018: ONE GENERATION REPRODUCTION S'I'UDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 3 1.6 + MEVALONIC ACID 4 2 + MEVALONIC - - _ _ _ . _ . . . _ _ _ . . . _ _ . . _ . _ . _ __ __ __ __ __ __ __ __ __ _._ __ __ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ -PR_ ECOHABIT_A_T-_I_O_N-_(DAY 1 OF STUDY TO -T. H_ E _D_ _A__Y_ OF _COH_ABI_TA_TIO~N):- ACID MAXIMUM POSSIBLE INCIDENCE 1176/ 28 FOUND DEAD 0 EXCESS SALIVATION 3/ 2 CHROMORHINORRIIEA 2/ 2 INCISORS: TOTAL MISSING/BROKEN MISALIGNED MOUTH: SCAB SWOLL,EN SNOUT TIP OF TAIL MISSING PTOSIS SOFT OR LIQUID FECES RALES RED SUBSTANCE IN CAGE LABORED BREATHING SCANT FECES LOCALIZED ALOPECIA: UNDERSIDE 7/ 2 7/ 2 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 I/ 1 o/ 0 o/ 0 o/ 0 8/ 1 RIGHT EAR SWOL,LEN 2/ 1 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 s t u d y . PROTOCOL 4 1 8 - 0 i a : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE m NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE i (PAGE 5): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP 2 3 4 DOSAGE MG/KG PFOS MEVALONIC ACID CONTROL 1 . 6 + MEVALONIC ACID 2 + MEVALONIC ACID ..--.--.._________._____________________-----.---..-.--------------- PRESUMED~ GEST~ATION_ : _ _ MAXlMUM POSSIBLE INCIDENCE 6091 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 PTOSIS 3/ 3 o/ 0 4/ 2 o/ 0 3/ 2 30/ 2 LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE NECK 33/ 3 lo/ 1 23/ 2 o/ 0 CHROMODACRYORRHEA 1/ la o/ 0 21 1 TIP OF TAIL MISSING o/ 0 o/ 0 221 1 DEHYDRATION o/ 0 o/ 0 51 1 RALES o/ 0 o/ 0 5/ 1 EMACIATION o/ 0 o/ 0 2/ 1 SCANT FECES o/ 0 o/ 0 I/ 1 MOIJTU: SCAB 3/ 1 4/ 1 o/ 0 SWOLLEN SNOUT o/ 0 I/ 1 o/ 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 1 0 9 2 6 was moribund sacrificed on day 1 2 of gestation; death was attributed to an intubation accident. b. Descriptions were (or were any combination of) red, dried, slight and moderate. # # Significantly different from the Group 1 value ( ~ ~ 0 . 0 1 ) . PKOTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 1 (PAGE 6): CLlNICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS SOFT OR LIQUID FECES LACTATION : 1/ la o/ 0 o/ 0 MAXIMUM POSSIBLE INCIDENCE 9 0 / 18 ?6/ 18 52/ 19 PTOSIS LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE o/ 0 o/ 0 5/ 1 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 1 0 9 2 6 was moribund sacrificed on day 12 of gestation; death was attributed to an intubation accident. I-RoroCoL 418-018: 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 PREC. O-H-A-B-I-T__A_TI__ON ~DS 1 OF STUDY pro THE DAY OF COHABITATION) : MAXIMUM POSSIBLE INCIDENCE 1150,' 28 1176/ 2 8 1176/ 28 MORIBUND SACRIFICED la 0 LOCALIZED ALOPECIA: TOTAL 6 2 LIMBS 5 2 NECK 2 0 UNDERSIDE 1 0 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 1 1 1 0 1 1 1 0 0 CHROMORHlNORRHEA 0 1 0 DECREASED MOTOR ACTIVITY la 0 0 PALE IN APPEARANCE la 0 0 COLD TO TOUCH la 0 0 SWOLLEN SNOUT 1 0 STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Rat 11562 was moribund sacrificed on day 16 of study. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 1 (PAGE a ) : CLINICAL OBSERVATIONS - SUMMARY - FO GENERATION FEMALE RATS CHALLENGE AND NOEL INVESTIGATION IN DOSAGE GROUP 5 DOSAGE MG/KG PFOS CHOLESTEROL CONTROL _ . - - - _ - _ _ _ _ _ _ _ . _ .._ ...... . . - - - - - - - - - - . - - . . . PRESUMED GESTATION: 1.6 + 6 CHOLESTEROL 7 2 + CHOLESTEROL MAXIMUM POSSIBLE INCIDENCE 601/ 27 6 0 4 / 28 MORTALITY 0 0 L,OCALIZED ALOPECIA: TOTAL NECK LIMBS UNDERSIDE BACK INCISORS: TOTAL MISSING/BROKEN MISALIGNED 77/ 5 22/ 1 59/ 4 14/ 1 o/ 0 32/ 2 o/ 0 32/ 2 52/ 5 o/ 0 41/ 4 2a/ 2 14/ 1 17/ 2 16/ 2 I/ 1 CHROMODACRYORRHEA CHROMORHINORRHEA CHEST OR LEFT FORELIMB: ULCERATION CHEST: SCAB -L.A. CTAT.I- O.-N : MAXIMUM POSSIBLE INCIDENCE 85/ 17 a6/ 20 651' 19 LOCALSIZED ALOPECIA: TOTAL LIMBS UNDERSIDE BACK RED PERIVAGINAL SUBSTANCE 16/ 4 12/ 3 4/ 1 o/ 0 INCISORS: MISALIGNED _.... 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. 18/ 5 15/ 4 4/ 1 3/ 1 o/ 0 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 - 6 2 9 5 . 2 5 ) 'I'ABLjE 2 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS ..._._...__......._____________ DOSAGE GROUP 1 8 9 10 11 12 13 DOSAGE MG/KG PFOS VEHICLE CONI'ROL 0.4 0.8 1 1.2 1.6 2 RATS EXAMINED a N 28 20 20 20 20 28 28 MORTAL1TY N 0 0 0 0 0 0 0 APPEARED NORMAL N 28 20 20 20 20 28 28 a. R e f e r to the individual clinical observations table (Table 2 3 ) for external observations confirmed at necropsy. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) MORTALITY N 1 1 MORIBUND SACRIFICED N lc 0 FOUND DEAD N 0 Id APPEARED NORMAL N 26 28 ESOPHAGUS : PERFORATION N lc 0 0 LEFT AXILLARY AREA: TISSUE DISCOLORED TAN N 1c 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 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 1 0 9 2 6 was moribund sacrificed on day 1 2 of gestation; death was attributed to an intubation accident. d. Rat 1 0 9 3 6 was found dead on day 2 of study. w +-- 0 418-018:PAGE B-11 la, 12 z z z z m 8 m U M .. u I n1 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 3 (PAGE 1 ) : TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY Fo GENERATION FEMALE RATS TERMINAL BODY WEIGHT MEAN+S.D. 417.5 5 18.3 399.8 2 25.2 409.8 rf 31.5 LIVER LIVER ( % ) MEAN2S.D. MEANkS .D . 14.3 f 0.7 3.4 f 0.2 14.9 5 1.9 3.7 2 0.5 15.3 5 1.2 3.7 rf 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 MEAN5S.D. 318.7 2 28.5 326.7 2 21.6 308.3 rf 42.0 309.5 5 19.8 305.6 2 23.2 298.6 rf 19.8 283.8 rf 7.5* LIVER MEAN2S.D. 13.2 5 1.8 13.8 2 1.5 13.6 5 1.1 13.7 2 1.4 14.5 rf 1.5 13.2 5 1.2 13.1 2 1.0 LIVER ( % ) MEANrfS.D . 4.1 5 0.5 4.2 + 0.3 4.5 5 0.5* 4.4 rf 0.5 4.8 5 0.3** 4.4 2 0.3 4.6 rf 0.3* ..._____._._____________________________-.--.----------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). * Significantly different from the Group 1 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value ( ~ ~ 0 . 0 1 ) . RATIOS ( % ) = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. rwmocoL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 3 (PAGE 2): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT - SUMMARY Fo GENERATION FEMALE RATS TERMINAL BODY WEIGHT LIVER MEAN+S . D . MEANtS .D. 416.6 24.6 15.3 2 2.1 LIVER ( 9 ) MEAN+S . D . 3.7 2 0.4 DELIVERED A LITTER _A- ND SACRIFICED ON DAY 5 OF LACTATION: ~ RATS TESTED N 18 393.7 2 30.4 16.2 2 2.3 4.1 2 0.4* 12 393.1 2 47.3 15.8 2 1.5 4.0 2 0.2 3 TERMINAL BODY WEIGHT MEANkS . D 319.5 t. 18.5 315.8 2 23.4 297.0 2 14.1 LIVER MEAN+S . D . 13.3 2 1.2 14.8 2 1.2** 13.8 2 1.6 LIVER ( % ) MEANkS . D . 4.2 2 0.3 4.7 2 0 . 3 * * 4.6 2 0.4* ._. _ _ _ . _ . _ _ . _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ~ ~ ~ ~ . ~ . ~ . . ~ ALL WEIGHTS WERE RECORDED IN GRAMS (G). * Significantly different from the Group 2 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 2 value ( ~ ~ 0 . 0 1 ) . RATIOS ( % ) = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 TERMINAL BODY WEIGHT MEANkS . D 441.0 5 30.6 LIVER MEAN+S . D . 14.1 2 1.3 LIVER ( % ) MEANkS . D . 3.2 0.2 _D_E_L_-I_V_ERE_D A_ L_ITT_ER_AN~D _S_A_CRIFICED ON DAY s _O_F_-_L_A_C_T__A_T_I_ON: RATS TESTED N 17 TERMINAL BODY WEIGHT MEAN2S.D . 328.0 2 31.9 LIVER MEAN+S . D . 13.4 2 1.4 LIVER ( % ) MEAN+S . D . 4.1 2 0.3 ALL WEIGHTS WERE RECORDED IN GRAMS (G). * Significantly different from the Group 5 value * * Significantly different from the Group 5 value (p.cO.05). (p.cO.01). 403.0 2 38.3* 14.3 2 0.8 3.6 2 0 . 4 * 387.5 2 31.3** 15.3 2 0 . 7 * 4.0 2 0.3** 13 7 303.9 A 19.0* 280.8 2 15.6** 14.3 2 1 . 7 12.6 2 0.9 4.7 2 0.5** 4.5 2 0.2* RATIOS ( % ) = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) x 1 0 0 . 8 , , 5 0 memW W * e 0 m N 'if d d d d ri +I +I +I +I +I +I m m in W e W * * in d m W m m r( m m in N N N N N N 0 r- W N W N 0 d N in in in ri d d ri ri ri +I +I +I +I +I +I W d in in d N in in m 0 W 0 d m e in in W N N N N N N m m d d ri e * * * d N w d r+iI d ri d d +I +I +I +I +I m ri N N W e *mN dPe ri m cr in in W N N N N N N m m m W m r- * 0 e in in m d ri d r- ri d +I +I +I +I +I + I N m 0m 00 * ri N N 0 P r- m e in W W N N N N N N N 0 N ri m m Lnl d d N in r- r- d d ri ri H ri +I +I +I +i +I +I N 0 in m m e * in m e N W e d m in in W N N N N N N r- 0 m m 0 m m i m ./ N -4 N in W wi ri d + I d H d d rii +I +I +I +I +I +I , 0 N in 6 e e 01 in W r- r- N d d m e in W r- N N N N N N m d m W in in 0 d N d +, *ri rd + I r- d m d +I +I +I +I e m in m m W in m d N N N m r- in N m e in W N N N N 9 9 9 a a a rA w cn [o ffl 1.3 - 5' 5' 5' 6' - w w w w W w I: z z P P 5: u b r i m in N m W 3: d N N m :: . w r 3 >I 3 * *% 2 2 2 3 4 2 a 0 a7 4 18-018:PAGE B- 15 I'KO'I'OCOL 418-018: ONE GENERATION KEPKODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEKOL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 4 (PAGE 2): BODY WEIGHTS - PREMATING - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GKOIJP DOSAGE MG/KG PFOS ...- .- - ..- - .- - - - - - . . RATS TESTED BODY WEIGHT (G) DAY 1 DAY 8 DAY 15 DAY 22 DAY 29 DAY 36 DAY 42b N MEAN+S . D . MEANkS . D . MEAN+S .D . MEAN2S.D . MEANiS .D . MEAN+S . D . MEAN+S .D . 216.2 2 11.3 229.6 2 11.0 240.2 2 14.1 249.1 +_ 13.6 257.4 2 14.6 265.4 k 17.1 267.6 2 17.4 215.6 2 12.1 231.6 2 13.3 [ 28la 240.5 2 17.5 [ 28la 249.5 2 18.7 [ 28la 253.5 2 19.7 [ 281a 258.1 2 19.5 [ 28la 261.3 2 23.1 [ 28la 215.6 2 11.6 234.2 2 11.5 242.9 2 11.8 252.6 2 13.4 258.2 5 14.2 263.1 2 15.6 266.4 2. 16.9 418-018:PAGE B- 17 I , ,, , NI , I I , c c c c c c m m m m m 0m 00N N v Ln v d 4 d rl d rl rl +I +I +I +I +I +i +I v 0 v) N in in m r- L n m N 0 N Ln d m v Ln Lo Ln m N N N N N N N I c c c c c c m m v Ln m P v 0 0 N v m Ln W rl rl d rl rl rl d m +I +I +I +I +I +I +I r- Ln m m rl v W 3 in rl 0 m m P r W rl m v v Ln Ln Ln 4 N N N N N N N 14 a: 0 I w W Ir, rl 3 z 0 2 H a C W 3 a u u 0 rl v m v m rl r- Ir, 0d v P 4 m d rl rl +I +i +I 2 mmm * v N N v iz rl rn cy in W 01 N N N N N u z H 2 b Wa: G 9 9 9 9 10 b a 3m : 10 ffl ffl ffl ffl H W ii5' 6' 5 w wWWW * a z z z II 0 m I m - w u - 2Pl b r l m in N m - 3: d N N :: v z W E * 2 a * 2 * d * 2 n Cmll a s 0 m PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 5 (PAGE 1): BODY WEIGHT CHANGES - PREMATING - SUMMARY - FO GENERATION FEMALE RATS RATS TESTED N 28 20 20 BODY WEIGHT CHANGE (G) DAYS 1 - 8 MEAN2S.D. +14.5 2 8.6 +21.2 6.6** t17.7 2 6.4 DAYS 8 - 15 MEAN,S.D. t8.7 2 5.2 +11.2 2 7.5 t11.6 2 5.2 DAYS 15 - 22 MEAN2S.D. t9.4 2 6.9 +10.0 2 6.8 +7.8 2 4.9 DAYS 22 - 29 MEAN2S.D. t7.4 2 5.Y +4.9 2 5.9 t4.6 2 6.8 DAYS 29 - 36 MEAN2S.D. t7.3 5 4.7 +9.0 5.7 t7.6 2 5.2 DAYS 36 - 42a MEAN2S.D. +3.0 2 8.4 t4.7 2 7.6 +1.6 2 9.1 DAYS 1 - 42a MEAN+S.D. +50.2 t 17.3 t61.0 2 15.0* +50.8 2 15.5 DAYS = DAYS OF STUDY a . L a s t value recorded before cohabitation. * Significantly different from the Group 1 value * * Significantly different from the Group 1 value (~~0.05). (p50.01). 20 20 28 28 +17.7 2 8.7 t18.3 2 9.1 t19.5 2 6.9* +15.4 2 6.6 t10.1 2 6.1 +9.0 2 6.4 t8.4 2 5.5 t8.2 2 6.6 +10.8 2 5.1 +9.0 2 5.0 t7.0 2 4.2 t7.1 2 6.6 +7.2 2 5.4 t6.5 2 6.3 t5.6 2 5.0 +2.8 2 4.9 t7.0 2 4.4 t6.7 2 4.0 t4.2 2 4.9* +4.2 2 4.2* +2.3 2 8.2 +0.2 2 8.5 -0.1 2 6 . 0 t1.5 2 6.8 +55.0 2 17.9 +49.8 2 14.1 t44.5 2 10.4 +39.2 2. 16.4* _ _ _ _ _ _ . -_ - -_ - - _ . _ . _ _ _ _ _ . _ _ _ _ _ _ _ _ _ - - - - _ - 418-018:PAGE B- 19 , m N 1 +i , Ni 1: c c N m N m P 0 m e in e m in +I +I +I +I +I +I W P P W m m m d + m + m + i+n e + m + I , 3in 2!W r-? W Er, z z 2 zw ; u 0 a !z i 5 m u z H 2 z Wa: % L m Wuz 3 0 s b 3: W x > n 0 m I N 9uW I in 2 W J m ** ** W W 6 P N m P W in e e W+I rc! m +I +I +I +I +i N m m 0 ri in N + m ri m + m + e + e + m + 0 P VI in m m m m in W m W m +I +' +I *I +I +' N e W m m 0 ri m r+ i 0 +r( m + m + m + N + f ' f ' f ' f 1 i'f i 9 a 9 9 9 9 in in m in m ffl 2 W W W W Wz z Wz z z z CA W i>n fi m in N m W ki m N N m e En ri m in N m W ri N N cl in in in in ;i) m 8 z * * * 2 tj a: >a:, E 2 2 V n n n n w 0 N x a " - - c m d 0 0. 0. a 00 n a, av i aV I a 2 !kj$ 3 iiii 0 w a> >n w ** +* N. N. N. W. W. r-wmmq +! +I +I +I +I . . . . . I D W ~ N O rmr-wm ri++++ 4 18-018:PAGE B-20 3KO , wi I ./ , di 0 rS4 c + i + I + i m. m. r . l . q . w .q . o i / mmwqmmril r i i +I +! +I +I +I +I +I 9 a h m. m . r .i m. m . r i. m . \ Ii u 2 i n m c o ~ c r o r ( m~ r i + + + + + q , + I w 0 W d h a c W +I + I 0 0 0 ri I - - - T! mind 0. 0. 0. - - - 0 0 0 aV I aV ! aV I @a,@ .i 2 2 2 ii ddd 0 m> m> m> - m m d 0 m ri q a ? a ? ? a m 'Z KO KO KO 5' 5' $WI Wi?w W P P z z m in N m W P N d NNm -7 dmm N W d N N m 2G l n PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) 'TABLE 6 (PAGE 1): MAI'ERNAL BODY WEIGHTS - GESTATION SUMMARY - F o GENERATION FEMALE RATS .~..._....__. DOSAGE GROUP 9 10 DOSAGE MG/KG PFOS 0.8 1 ..._._....________._____ RATS TESTED N 28 20 20 28 28 PREGNANT N 27 20 17 19 18 25 27 MATERNAL BODY WEIGHT (G) DAY 0 MEAN2S.D. 275.0 2 21.6 281.6 k 17.0 274.0 2 22.1 274.2 2 19.9 268.2 2 18.6 267.8 2 15.8 259.7 2 19.0* DAY 1 MEAN2S.D. 283.3 2 20.4 291.2 2 18.5 281.1 2 23.3 282.6 2 21.1 275.4 2 17.5 272.6 2 16.8 265.7 2 19.9** DAY 2 MEAN2S.D. 287.9 2 20.9 294.6 2 19.7 284.4 2 23.2 286.6 2 22.0 277.6 2 17.8 275.0 2 16.2* 267.0 2 17.9** DAY 3 MEAN2S.D. 289.0 2 20.2 296.2 t 18.6 284.7 2 22.6 287.6 2 19.8 277.5 5 19.4 275.4 2 16.1* 268.0 2 1 7 . 1 * * DAY 4 MEAN2S.D. 291.7 k 19.5 299.6 k 19.7 286.6 5 22.9 288.2 2 18.1 278.6 2 18.6 276.5 _t l6.5* 269.4 2 1 7 . 0 * * DAY 5 MEAN+S.D. 293.6 2 19.9 304.0 t 19.8 290.4 2 23.7 291.2 2 18.6 281.3 2 16.8 277.9 2 1 7 . 0 * * 271.8 2 1 7 . 0 * * DAY 6 MEAN2S.D. 295.4 2 20.2 306.2 2 20.1 289.8 2 22.0 292.1 2 18.5 283.6 2 18.0 280.2 2 16.9* 272.9 2 17.1** DAY 7 MEAN2S.D 298.9 2 20.0 308.4 f 19.2 292.5 2 21.7 294.3 2 19.9 286.1 2 19.2 283.6 2 1 6 . 6 * 276.5 2 1 7 . 6 * * DAY 8 MEAN+S.D. 299.6 2 21.8 311.8 2 19.1 295.2 2 21.8 296.8 2 16.5 289.5 2 19.9 286.1 2 15.9* 278.7 2 18.3** DAY 9 MEAN+S.D. 304.0 2 21.9 314.8 2 19.3 289.9 2 22.3 300.2 2 16.9 291.4 2 19.3 288.4 2 19.0* 281.5 2 17.4** DAY 10 .- - - - - - MEAN2S.D. 308.4 5 21.9 318.7 2 19.5 303.5 2 DAY = DAY OF GESTATION * Significantly different from the Group 1 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value ( p ~ O . 0 1 ) . 23.0 303.9 2 17.0 295.3 2 20.3 293.1 2 17.8* 286.2 2 17.8** . _ _ _ _ . _-_ - _ _ - - ___ _ _ __ _ __ _ _._ _ _ _ _ __ _._ __ . _ _ _ _ _ __ __ _._ _ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 6 (PAGE 2): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - FO GENERATION FEMALE RATS .. DOSAGE GROUP 1 8 9 10 11 12 13 DOSAGE MG/KG PFOS VEHICLE CONTROL 0.4 0.8 1 1.2 1.6 2 RATS TESTED N 28 20 20 20 20 28 28 PREGNANT N 27 20 17 19 18 25 27 MATERNAL BODY WEIGHT (G) DAY 11 MEAN2S.D. 313.8 2 21.1 324.2 2 19.5 308.6 2 23.2 310.0 2 17.9 302.2 2 19.9 296.4 2 18.4** 291.2 5 19.4** DAY 12 MEAN2S.D. 318.2 2 22.3 329.8 2 19.2 312.9 2 24.7 312.4 2 18.5 306.8 2 22.5 299.1 2 19.2** 294.4 5 18.8** DAY 13 MEAN2S.D. 322.8 2 22.4 332.0 20.2 317.2 2 25.4 318.3 2 17.2 309.3 2 19.7 304.3 2 19.4** 299.2 2 18.4** DAY 14 MEAN2S.D. 326.8 2 22.6 337.6 2 21.2 323.4 2 27.3 326.6 2 17.9 316.2 2 19.4 310.4 2 18.2* 302.9 2 19.0** DAY 15 MEAN5S.D. 333.9 2 23.0 347.1 2 20.9 332.4 2 25.3 333.1 2 18.3 322.4 2 21.2 319.1 2 19.0* 313.3 2 19.4** DAY 16 MEAN2S.D. 343.1 5 24.1 357.0 2 22.6 343.3 2 26.0 344.8 2 16.8 333.3 5 21.3 328.7 2 19.6* 323.4 2 19.3** DAY 17 MEAN2.S.D. 356.0 2 26.6 370.4 2 23.0 357.0 2 26.4 357.9 2 18.1 347.9 2 22.5 343.4 2 20.6 337.3 5 20.0** DAY 18 MEAN2S.D. 370.1 28.6 383.6 2 23.9 371.2 2 26.1 373.4 5 18.7 362.7 2 23.7 358.5 2 20.3 352.1 2 21.0* DAY 19 MEAN2S.D. 381.9 2 29.9 395.8 2 24.4 384.6 2 28.0 386.7 2 17.1 373.2 2 25.3 373.0 2 21.8 365.6 2 21.5* DAY 20 MEAN2S.D. 396.2 5 31.3 410.3 2 26.0 395.6 2 26.2 400.2 2 20.1 387.1 2 27.0 385.1 2 25.0 380.0 5 23.3 DAY 21 MEAN2S.D. 410.6 5 31.1. 421.2 2 28.9 408.5 2. 25.3 408.6 5 22.2 394.5 2 31.7 398.3 2 26.1 397.5 2 30.2 [ 18la [ 121a [ 12la [ 15la [ 20la [ 21la ...__._..._.___.____.~.....~~~~..~~..~~~~.~~~~~~~~~~~~ DAY = DAY OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes values for darns that delivered. * Significantly different from the Group 1 value * * Significantly different from the Group 1 value (~~0.05). (p<O.Ol). PKO?'OCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 6 (PAGE 3): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - FO GENERATION FEMALE RATS PREGNANT N 27 MATERNAL BODY WEIGHT (G) DAY 0 DAY 1 MEAN+S . D . MEANLS .D . 275.3 2 19.0 283.0 5 18.8 DAY 2 MEAN2S.D . 286.3 2 18.9 DAY 3 DAY 4 MEANkS . D . MEANLS . D . 287.0 5 18.9 289.0 5 18.9 DAY 5 MEANLS . D . 290.8 5 19.3 DAY 6 MEANkS . D . 292.9 19.0 DAY 7 MEANkS .D . 296.0 5 17.9 DAY 8 MEANkS . D . 299.0 5 18.2 DAY 9 MEAN2S.D . 302.6 5 19.4 DAY 10 MEANLS .D . 305.9 5 19.0 DAY = DAY OF GESTATION d . Excludes rat 10936, which was found dead prior to gestation. * Significantly different from the Group 2 value (pzO.05). * * Significantly different from the Group 2 value (~50.01). 26 266.6 2 22.9 271.5 5 22.3 273.4 2 22.4 273.8 2 23.1 275.6 5 22.2* 275.9 5 22.1** 277.9 5 23.3** 280.0 2 22.3** 282.3 2 22.9** 285.6 5 23.1** 289.5 2 2 3 . 4 * * 27 272.0 2 16.9 275.7 2 17.5 278.1 3 18.5 278.6 2 19.0 279.1 5 18.4 281.3 2 18.5 282.4 2 18.9 284.7 2 19.9* 286.6 2 18.9* 290.0 5 20.5* 294.6 2 19.7* PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 6 (PAGE 4): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - FO GENERATION FEMALE RATS ...- - .- .- .- .. .- ... .- .. .....................- .. .- - - .... .... ...- - .- - - - - - - - . DOSAGE GROUP 2 DOSAGE MG/KG PFOS MEVALONIC ACID CONTROL RATS TESTED N 28 PREGNANT N 27 26 27 MATERNAL BODY WEIGHT ( G ) DAY 11 MEANAS.D . 311.7 A 18.5 294.4 2 24.9** 300.7 2 20.5 DAY 12 MEAN+S . D . 313.5 18.3 299.0 2 25.2* 303.3 2 21.0 DAY 13 MEANAS .D . 318.3 2 18.6 302.9 2 25.9* 309.1 5 20.5 DAY 14 DAY 15 DAY 16 DAY 17 DAY 18 MEAN+S .D . MEANkS . D . MEAN+S .D . MEANAS.D . MEAN+S . D . [ 26lb 322.8 2 18.8 [ 26lb 330.6 2 18.8 [ 26lb 339.4 2 18.8 [ 26lb 351.0 2 20.7 26lb 363.4 2 20.8 308.4 2 26.9 315.9 2 25.8 325.7 2 27.2 340.5 2 27.6 356.4 2 28.9 312.2 2 21.6 319.4 2 24.0 329.9 5 25.6 344.2 2 25.8 359.8 2 27.1 DAY 19 MEANkS . D . [ 26lb 379.4 2 24.9 368.9 2 29.9 372.4 2 26.8 DAY 20 MEANkS . D . [ 26lb 390.9 2 24.7 382.7 2 32.6 385.3 28.4 DAY 21 MEANAS. D . [ 26lb 403.5 2 24.8 398.2 2 34.1 403.4 2 35.1 .___ [ 25lb,c [ 25lc [ 19lc ___---___-____________________ DAY = DAY OF GESTATION [ I = NUMBER OF VALUES AVERAGED a. Excludes rat 10936, which was found dead prior to qestation. 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 (~50.05). * * Significantly different from the Group 2 value (p50.01). PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 KATS TESTED N 2 la PREGNANT N 25 28 27 MATERNAI, BODY WEIGHT (G) DAY MEANtS.D. 284.8 2 22.3 265.1 2 17.9** 264.0 k 17.9** DAY MEANjS . D . 292.6 2 22.7 268.2 2 19.4** 267.1 2 18.3** DAY MEAN+S .D . 297.6 2 21.7 271.5 2 18.7** 270.4 2 17.7** DAY MEAN+S .D . 299.4 5 22.6 272.6 2 18.5** 270.1 2 15.4** DAY MEAN+S . D . 302.3 5 21.4 274.2 2 18.3** 271.4 2 17.3** DAY MEANkS . D . 305.7 2 22.4# 277.2 2 19.5** 273.4 2 1 6 . 7 * * DAY MEAN+S . D . 306.6 2 25.2 279.6 2 19.5** 276.0 2 16.4** DAY MEAN+S . D . 309.8 t 24.0 281.5 2 20.5** 278.4 2 17.7** DAY MEAN+S . D . 315.1 2 25.0# 283.8 2 21.0** 282.2 2 17.5** DAY MEANzS . D . 317.7 2 24.6# 287.8 2 19.6** 284.9 2 17.2** DAY .- - - - - .. MEANkS.D . 323.6 2 24.4# - . . . . - - . _ _ _ _ __ _ __ _._ . ._ _._ . _ __ __ __ _ _ _ - - - - - _ _ _ _ _ _ _ DAY = DAY OF GESTATION a . Excludes rat 11562, which was moribund sacrificed prior to gestation. it Significantly different from the Group 1 value (~50.05). * * Significantly different from the Group 5 value ( p 5 0 . 0 1 ) . PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 6 (PAGE 6): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS PREGNANT N 25 MATERNAL BODY WEIGHT (G) DAY 11 DAY 12 DAY 13 DAY 14 MEAN+S . D . MEANAS.D . MEAN+S . D . MEANkS .D . 329.0 2 24.2# 332.9 f 25.1# 335.7 2 26.0# 340.7 2 25.1# DAY 15 DAY 16 DAY 17 DAY 18 DAY 19 MEANtS.D . MEANAS.D . MEANAS.D . MEANLS.D . MEANkS . D . 347.6 2 24.4# 357.3 26.8# 370.2 f 26.2# 383.2 2 27.0 397.1 2 28.6 DAY 20 MEAN-iS.D . 410.3 2 29.6 DAY 21 MEANAS.D . 428.0 31.1# [ 24lb DAY = DAY OF GESTATION [ 1 = NIJMBER OF VALUES AVERAGED a . Excludes rat 11562, which was moribund sacrificed prior to gestation. b. Excludes values for dams that delivered. # Significantly different from the Group 1 value (p50.05). * * Significantly different from the Group 5 value (p<O.Ol). 28 295.8 2 21.4** 300.8 2 22.6** 305.6 2 22.1** 310.1 2 23.8** 318.4 2 24.9** 329.0 2 26.4** 342.4 2 26.8** 357.2 5 29.2** 370.1 _+ 31.7** 383.6 2 32.6** 392.0 2 31.6** [ 23lb 27 292.8 2 1 8 . 7 * * 296.3 k. 20.4** 300.8 2 21.5** 305.4 2 2 1 . 0 * * 314.4 21.9** 325.4 2 23.2** 339.7 2 24.1** 354.8 2 24.9** 369.3 2 26.2** 380.9 + 28.1** 394.5 f 29.0** [ 19lb 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 MEAN5S.D. +23.8 _f 7.0 + 2 6 . 8 _f 7.5 +18.5 5 8 . 6 * ~ 2 0 . 05 7.0 +17.9 2 6.2* +15.8 2 5.3** +16.8 2 6 . 0 * * DAYS 7 - 14 MEAN5S.D. -127.95 6.1 +29.2 5 8.7 +30 9 5 8.5 +32.3 2 7.5 +30.1 2 6.9 + 2 6 . 7 2 6.4 +26 DAYS 14 - 21 MEAN5S.D. +83.8 5 13.3 +84.3 2 13.6 +86 2 5 7.6 +78.4 2 25.6 DAYS 0 - 21 [ 18la 121 a [ 12la MEANiS.D.+135.5 5 15.6 +139.7 % 17.5 +133.4 2 13.7 +132.6 2 29.6 [ 18la 12la I 12la ..................................................................................... DAYS = DAYS OF GESTATION [ 1 = NUMBER OF VALUES AVERAGED a. Excludes values for darns that delivered. * Significantly different from the Group 1 value * * Significantly different from the Group 1 value (~~0.05). (pc0.01). +79.9 2 16.6 [ 151a t128.3 + 23.5 +87 2 2 15.3 201 a +129. +96 418-018:PAGE B-28 , NI , 8 8 , c c r . n . ~. m. r-mm d$ , o~ NUrnU I +I +i +;I +I= ; r( dl aid u 3 d .i 9 0 u a u 3 P .idr u u id ffl id 3 a u id a c -rl u id u 63 a, tn w 0 a nnn z -a ` a Co auJ .i -4 c1w id -i uh d mu 0 wid tnffl 0 VI 0.0 ur: w h a 0 .ri 3 -r( ii m i i o ri > C b E am .N a5 3 $ 3 d dU d3 -ad 03 w aJ0 > -A u h raiJ w aa r- -r r( d d N N c 0r 0 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE: -7 (PAGE 3 ) : MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL 6 1.6 + CHOLESTEROL 7 2 + CHOLESTEROL RATS TESTED N 2 7a 28 28 PREGNANT N 25 28 27 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 MEAN+S .D . +25.0 + 10.4 DAYS 7 - 14 MEnrJ+S . D . +30.9 2 6.9 DAYS 14 - 21 DAYS 0 - 21 MEAN+S . D . MEANkS .D . 186.5 + 14.9 I 24lb +142.8 2 19.8 [ 24lb ___________.____..______________________-~---------.--------- 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 ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 5 value ( ~ ~ 0 . 0 1 ) . m m d d r- r- d d m m Ti d , I c , , Od 0 m m d N d d r- r- ri d me 0 0 0 N N N 0 zcri 0 a E+ 0 U d am a W d d d * N Ln d N r m + I +I; . - 0 m d m m m N N 0 0 N +I N m m N d 0 N +I ri m m N ul ul d +I ** m 0 0 m 0 W 0 * N N * +I + I 0 m * m 0 0 m m m W m d d N +i +I 0 r- d W d N m m 0 in W m N N +i +i m r d m 0 d m m 9 a z z 2 10 i' a W W b z W 5: - E; U - cri E 5 9 2 2 z > n 4 E + d in W b n 3: U in W H ir: W 2 in c? 8 W 4 3 nW 0 m G 0 -4 u m u U m -3 w 0 ul x a 0 u h .4 h a z m a 3 a >-4 ? .ri I 0Gin 0 0' uo m VaI a1 a2 ffld or6 .wr( ? .4 d h oa am o1 fflu fflm 4 18-018:PAGE B-30 0 0 N 9 m +I d d m 0 0 m W m N * m m +I ri ri d in 0 ri m m m Ln v m N d m m +I +i d d m in m m m d N 0 6` 5` 9 n z z m (0 w z 2 8 - wGl m 0) 0 a D-r W 1 z Ln H 5 *2 aw z a 3 u w 4 U a: c4 E 4 18-018:PAGE B-3 1 , * W N N m m +I ri ri v I +i m m I Ni N , -c m v N 0 0 +I N N m m-? N I rll I m v N r r +I i( ri v N ri m a i' z z rn W z - e: E - E; W 4 ki q a: bz " a W W H l3 2 2 W r We: ii W n 0 a a m d 0 -4 u m u u rd 4 w 0 m h a u 0 !-! .r( !a-! m c'3a oa -4 utn rdG u .r( Ua ? .4 ci> !-! . . u0rn3 !o-! c0m- -r i -4 0 0 !-!0. ' a42 0 0 a a, aV l aV I a, 2 - - Ua"aJ Gag3 .d a , 4 4 ! U -!u .4> a> m rdw 6') -d m m a !-! u a a 63 r6n 3o o3 fi"l$l$ 418-018:PAGE B-32 E'KOTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABL,E 9 (PAGE I): BODY WEIGHT CHANGES - LACTATION - SUMMARY - FO GENERATION FEMALE RATS PREGNANT N 19 20 17 19 18 17 19 DELIVERED A LITTER N 19 20 17 19 18 17 19 INCLUDED IN ANALYSES N 19 20 17 18a 18 14a 6a BODY WElGHT CHANGE ( G ) DAYS 1 - 5 MEANtS.D. +17.4 11.1 +15.7 2 15.4 +4.3 5 25.8* + 4 . 0 2 10.5* + 7 . 5 5 11.6 il.1 5 12.7** - 3 . 5 2 16.8* __..__._________________________________--..--..--------.---.--- DAYS = DAYS OF LACTATION a. Excludes values for darns that were sacrificed due to no surviving pups prior to day 5 of lactation. * Significantly different from the Group 1 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value ( ~ ~ 0 . 0 1 ) . 4I 8-018:PAGE B-34 m d I .d u m u U rd rl 6 m d ri I m m ri rl , z z g 9--u Ll W (0 W 3: U w 2 a W w 5 z a 5 c7 3 4 > w U a 0: a H z 0 m w z 8 H 2:: $ H 4 W 0 z Ei z;uW ;i 4 $ U 4 0 8E+ m W J 0 3: V ..a H U A 2 z U H 0 J z ffl- om hN PI. g &L1 hm ON W 3 E- 2.r .. z Xw om z 5 u z 22 8> aw k r.2 d z -L'i ox HO wa 32 u r.2 Wb 83 m d 0 m d v J 0 V 0 b 0 d a 8 I 8 , I, W m m +! rl 0 m v m d 0 +I N wi r-$ +i r- rl z z d W EJ A z a W p: W ;i W n 418-018:PAGE B-35 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 10 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PREMATING - SUMMARY - FO GENERATION FEMALE RATS FEED CONSUMPTION (G/DAY) DAYS 1 - 8 DAYS 8 - 15 MEAN2S.D. MEAN+S.D. 19.6 2 3.0 [ 26la 19.0 1.8 DAYS 15 - 22 MEAN2S.D. 18.9 2 2.3 DAYS 22 - 29 MEAN2S.D. 18.8 2 2.0 DAYS 29 - 36 MEAN2S.D. 18.8 2 2.1 DAYS 36 - 42b MEAN2S.D. 18.3 2 2.1 DAYS 1 - 42b MEANcS.D. 18.9 2 1.8 19.6 2 1.9 [ 19la 19.0 2 3.6 19.4 2 1.8 19la 19.4 2 2.0 20.2 2 2.2* [ 17la 19.4 5 2.1 19.6 5 1.8 19.1 2 1.9 19la 18.9 T 1.5 [ 19la 19.2 2 1.6 [ 18la 19.0 f. 2.5 [ 19la 18.3 5 2.1 [ 18la 18.2 f. 2.6 19.0 2 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 ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value ( ~ ~ 0 . 0 1 ) . 18.9 2 2.2 19.4 2 2.9 19la 20.4 f. 2.2* 19.8 5 1.8 19.7 5 2.2 191 a 18 3 5 2.0 191 a 19 4 5 1.8 191 a 19.2 5 1.5 18.7 5 1.8 19.1 2 1.4 [ 17la 18.8 2 1.7 [ 1913 18.3 2 1.7 [ 19la 17.4 f. 2.2 [ 191a 18.6 5 1.3 [ 19la 13 2 28 19.9 2 2.0 18.9 5 1.9 19.1 5 2.1 [ 27la 18.5 2 1.6 [ 27la 18.3 5 2.2 17.0 5 1.6* 18.7 2 1.6 19.0 2 1.8 [ 27la 18.0 2 1.5 18.2 2 1.6 [ 27la 17.6 2 1.7* 17.0 5 1.5** 26la 16.3 5 1.6** [ 27la 17.8 2 1.3* [ 271a 418-018~PAGEB-37 * * * * i m. i. w r m m i n i i N P i a i aN a N i i m +IT +I +IF+lG + I 2 +I +I I N N NNN t . n- m. o. Y m. - m. - m. r. -, m m i i mmmr-m I r i d d r i i l * * I * + * I w m WmNmOi . . . . ./ dmNNNNNi m RP P RB m +I +! N NN N NI N 0 m.I 0_ - m. m. u m _ - , I mmm m m r m i i i iiridril m N i N Q m +I2+I N N w 0 0 i N N 5' ii5' 5' 5' ? 9 ? 9 9 9 n z m m (0 10 m cii m z w w E W z E $w I z W z W z m W m 2 m m N m 10 d u N N 2 iN N m e m E a ri m in N m 10 i ri N N m n W L? L J m m ro m 10 3 b b b $I z z z F.2 E 2 2 2 V n n d n a - -. . mi 0. 0. - -0 0 aV I aV I :; r i d i>d m> PROTOCOL 418-018: 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 FEED CONSUMPTION (G/DAY) DAYS 1 - 8 MEAN+S . D . DAYS 8 - 15 MEAN+S .D . DAYS 15 - 22 MEANAS.D . DAYS 22 - 29 MEAN+S . D . DAYS 29 - 36 MEANiS . D . DAYS 36 - 42C DAYS 1 - 42C MEAN+S . D . MEAN+S . D . 19.1 + 1.5 19.7 5 1.7 [ 27lb 18.7 2 1.6 18.6 2 1.3 18.8 2 2.1 17.9 2 1 . 5 * * 18.3 5 1.4** [ 2313 17.3 2 1 . 5 * * 18.3 2 1.2** DAYS = DAYS OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Excludes values t.hat were associated with spillage. b. Excludes values for rat 11562, which was moribund sacrificed on day 16 of study. c. Last value recorded before cohabitati.on. # Significantly different from the Group 1 value ( ~ ~ 0 . 0 5 ) . # # Significantly different from the Group I. value ( ~ ~ 0 . 0 1 ) . * * Significantly different from the Group 5 value ( ~ ~ 0 . 0 1 ) . 19.0 2 1.5 18.9 5 1.9 18.9 5 2.1 [ 27la 17.7 2 1.9** 18.2 $ 2.4** [ 25la 17.3 2 1.6** 18.4 2 1.6** PROTOCOL,418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 11 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PREMATING - SUMMARY - FO GENERATION FEMALE RATS RATS TESTED N 28 FEED CONSUMPTION ( G / KG/DAY) DAYS 1 - 8 DAYS 8 - 15 MEAN2S.D. MEAN+S.D. 88.2 f 14.0 [ 26la 80.9 f 4.9 DAYS 15 - 22 DAYS 22 - 29 MEAN+_S.D. 77.7 2 6.3 MEAN+S.D. 74.9 + 5.3 DAYS 29 - 3 6 MEAN2S.D. 72.6 6.4 DAYS 36 - 42b MEAN+S.D. 69.1 f 5.4 DAYS 1 - 42b MEAN5S.D. 77.2 f 4.6 20 87.0 L 8.3 [ 19la 78.4 2 14.4 77.1 f 6.8 19la 74.6 f 6.2 76.6 5 6.9 [ 17la 70.7 2 5.3 17.6 f 5.8 20 85.3 + 7.8 [ 19la 79.4 + 4.9 [ 19la 77.4 + 3.8 [ 18la 74.5 2 7.4 [ 19la 70.6 + 5.2 [ 18la 68.6 2 7.6 76.7 f 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 ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value (p50.01). 20 84.8 + 8.9 81.6 f 10.5 [ 19la 82.7 2 8 . 8 * 77.4 2 6.9 75.1 2 7.8 [ 19la 68.8 2 6.7 I 19la 78.8 + 6.3 [ 19la 20 85.8 2 6.6 78.6 + 4.9 77.9 2 3 . 4 [ 17la 74.1 2 6.5 [ 19la 70.2 2 5.9 [ 19la 65.8 2 6.7 [ 19la 75.4 2 4.0 [ 191a 28 28 88.4 2 7.7 78.9 2 5.6 77.3 2 5.3 [ 27la 73.2 2 4.4 [ 27la 70.8 2 6.8 65.2 2 4.4* 75.8 f 4.2 85.0 2 7.0 [ 27la 76.6 2 4.7* 75.0 f 5.2 [ 27la 71.1 f 6 . 0 * 68.2 4.9* [ 261a 63.9 f 4.3** [ 27la 73.4 2 3 . 8 * [ 27la wwI \o PRorocoI, 418 018: ONE GENERATION REPRODUCTION s m x OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLjE 11 (PAGE 2): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PREMATING - SUMMARY - F O GENERATION FEMALE RATS INCLUDED IN ANALYSES N 28 2 8a FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 MEAN5S .D . DAYS 8 - 15 MEAN+S . D . DAYS 15 - 22 MEANkS .D . DAYS 22 - 29 MEANkS .D . DAYS 29 - 36 DAYS 36 - 42C DAYS 1 - 42C MEAN+S . D . MEANkS . D . MEANkS .D . 89.5 2 6.9 [ 26lb 85.6 2 6 . 6 # # 82.4 2 7.0# 27lb 78.8 2 6.2# 74.6 2 6.5 [ 26lb 70.4 5 6.8 80.3 5 4.5# 85.8 2 7.8 80.4 2 12.4* 76.9 2 7.9** [ 27lb 71.6 2 6.2** [ 27lb 71.9 2 6.3 67.3 2 5.6 [ 27lb 75.7 2 5.3** [ 27lb DAYS = DAYS OF STUDY [ 1 = NUMBER OF VALUES AVERAGED a. Excludes values for rat 10936, which was found b. Excludes values that were not recorded or were c. Last value recorded before cohabitation. # Significantly different from the Group 1 value # # Significantly different from the Group 1 value * Significantly different from the Group 2 value * * Significantly different from the Group 2 value dead on day 2 of study. associated with spillage. (~~0.05). (~~0.01). (~~0.05). (~~0.01). 28 86.8 5 5.6 [ 27lb 78.9 2 7.7** 76.9 5 7.5** [ 27lb 71.8 2 5.6** [ 26lb 70.9 5 10.4 [ 26lb 65.6 2 6.5** 75.4 2 4.7** PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 11 (PAGE 3 ) : RELATIVE FEED CONSUMPTlON VALUES (G/KG/DAY) - PREMATING - SUMMARY - FO GENERATION FEMALE RATS FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 MEAN+S .D . 85.2 2 5 . 1 83.6 5 6.0 DAYS 8 - 15 DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 42C DAYS 1 - 42C MEAN+S . D . MEAN+S . D . MEAN+S . D . MEANkS .D . MEANrf.SD.. MEAN+S . D . 80.8 2 5.2 [ 27la 80.2 2 6.9 [ 27lb 76.2 5 6.3 [ 27lb 77.9 rf. 8 . 0 # # [ 25la,b 71.9 rf. 6.5 [ 27lb 78.6 5 4.7 [ 27lb 79.0 2 4.0 76.7 2 6.9 71.4 2 3.9** 71.5 2 3.6** [ 23la 67.2 rf. 4.5** 75.1 2 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. itit Significantly different from the Group 1 value ( ~ ~ 0 . 0 1 ) . * Significantly different from the Group 5 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 5 value ( ~ ~ 0 . 0 1 ) . 84.6 rf. 6.3 79.4 2 7.2 76.9 rf. 8.8 [ 27la 70.3 2 6.6** 71.4 2 7 . 7 * * [ 25la 67.6 2 6.5** 75.5 2 5.7* PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACIDICHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 12 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP 1 10 DOSAGE MG/KG PFOS VEHICLE CONTROL . _ ^ - - - - . - - - _ _ . _ _ . . . . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - . RATS TESTED N 28 PREGNANT N 27 20 17 19 18 25 27 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 7 MEAN2S.D. 22.3 2 2.2 22.7 2 2.3 20.6 2 2.4* 20.7 2 2.0* 19.6 5 1.9** 19.0 2 2.1** DAYS 7 - 14 DAYS 14 - 21 DAYS 0 - 21 MEAN2S.D. 22.9 2 2.2 MEAN2S.D. 24.3 2 2.2 MEAN2S.D. 23.2 2 1.9 23.2 2 1.6 1 19la 24.5 2 2.5 23.6 2 1.8 22.7 2 2.5 25.1 2 2.2 1 13la,b 22.8 2 2.2 21.7 2 2.7 25.0 2 2.5 22.5 2 1.8 22.0 f. 2.1 24.7 f. 2.6 22.1 f. 1.9 21.2 2.7 25.2 5 2.3 1 23lb 21.8 2 2.1* [ 13la,b [ 23lb . _ - - _ _ _ _ _ _ _ _ _ _ _ __ __ __ _._ _- _-_ . - _._ - - _ -_ - - _-_ - - _-_ - . -.-_ -_ - _-..- _ _ . _ _ __ __ __ __ __ __ _ _ _ _ _ _ _ _ _ _ _ DAYS = DAYS OF GESTATION [ 1 = NUMBER OF VALUES AVERAGED a. Excludes values that were not recorded as well b. Excludes values for dams that delivered. * Significantly different from the Group 1 value * * Significantly different from the Group 1 value as those associated with (~~0.05). (~~0.01). spillage. PROTOCOL 418-018: 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 PREGNANT N 27 26 27 MATERNAL FEED CONSUNPTION (G/DAY) DAYS 0 - 7 MEAN+S . D . 21.6 + 1.8 18.2 2.3** DAYS I - 14 MEAN+S .D . 23.2 2 2.1 20.9 2 1.9** DAYS 14 - 21 MEAN+S . D . 24lb,c 24.4 + 2.5 25.0 2 2.4 DAYS 0 - 21 MEANkS.D . [ 26lb 23.0 + 1.8 [ 22ld 21.5 2 2.0** [ 26lb [ 22ld .________._.__._._______________________.~~~~-~-~~~~~ DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Excludes rat 10936, which was found dead prior to gestation. b. Excludes values for dam 10926, which was moribund sacrificed on day 12 of gestation (death was attributed to an intubation accident). c. Excludes values that were associated with spillage. d. Excludes values for dams that delivered. * * Significantly different from the Group 2 value ( ~ ~ 0 . 0 1 ) . 418-018:PAGE B-44 * *c c i c I ! 0mmml . . . I N r ai N v d l UI r +I +ID +ID +ID N N N Nl m molni .I .I . I / m Olnril r- N N N i, * * i c c i e , N r . i .d m. i / N N aN u r i I UI m +I N m a ri u.r( id u rn a, ts, mr m w - N N N N QnUu 0 u il 0 d .d 0 - - - - ln +lm +I+ +lm +lm N NN" mm+m . I . I . c _ .Y N N -2> a z 10 \ W - 6`W z I: 0 H f10 8u r n W u G 0 5 i 810 2 W h Wua b 2 3r PROI'OCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 1 3 (PAGE 1 ) : MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY - FO GENERATION FEMALE RATS 20 20 20 28 28 20 17 19 18 25 27 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 MEAN2S.D. 7 7 . 2 2 5 . 7 76.3 2 5.0 72 2 2 5.4* 72.3 2 7.2* DAYS 7 - 1 4 DAYS 1 4 - 2 1 DAYS 0 - 2 1 MEANiS.D. 7 3 . 4 2 4 . 2 MEAN2S.D. 6 6 . 8 2 5 . 4 MEAN2S.D. 7 1 . 9 5 4 . 0 72.2 5 4.1 [ 19la 65.1 2 6.8 [ 18lb 7 0 . 8 2- 4 . 0 [ 18lb 74 0 2 4 . 2 68 8 2 4 . 3 llla,b 70 5 2 3.9 111a , b 70.6 2 8.3 68.1 2 9.0 [ 12lb 71.2 2 6 . 9 12lb [ I = 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 ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value (pc0.01). PRorocoL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 5 6.1 66.3 2 6.4** 68.4 2 8 . 2 * * DAYS 7 - 14 MEAN+S . D . 74.9 _t 6.0 71.5 2 3.4* 70.1 2 6.4** DAYS 14 - 21 MEAN_tS.D . 24lb,c 67.9 5 4.8 71.8 2 5.9 [ 25lc 68.6 8.4 DAYS 0 - 21 MEANkS .D . [ 25lb,c 72.5 2 4.0 [ 2lld 70.2 2 4.2 [ 19ld 70.0 2 5.9 [ 25lb,c [ 21ld [ 19ld ..._____________________________________-.------------------ DAYS = DAYS OF GESTATION [ 1 = NUMBER OF VALUES AVERAGED a. Excludes rat 10936, which was found dead prior to gestation. b. Excludes values for rat 10926, which was moribund sacrificed on day 12 of gestation (death was attributed to an intubation accident) . c. Excludes values that were associated with spillage. d. Excludes values for dams that delivered. * Significantly different from the Group 2 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 2 value (p.cO.01). PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 13 (PAGE 3): MATERNAL,RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY - FO GENERATION FEMALE RATS MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 MEAN+S . D . 76.7 + 7.8 69.1 f 6 . 0 * * 67.5 f 7.2** DAYS 7 - 14 MEAN+S .D . [ 23lb 74.8 2 6.7 71.8 + 5.1 71.8 f 5.3 DAYS 14 - 21 MEAN+S .D . [ 24lb 66.9 f 6.4 [ 27lb 71.2 f 4 . 4 * [ 26lb 72.6 f 6 . 0 * * DAYS 0 - 21 MEAN+S .D . [ 23lc 72.2 + 5.7 [ 23lc 70.6 3.5 [ 191c 70.4 2 5.3 [ 23lc [ 23lc [ 191c ________.__.____._______________________-------.-------------- 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 (pc0.05). * * Significantly different from the Group 5 value ( ~ ~ 0 . 0 1 ) . PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 14 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - LACTATION - SUMMARY - FO GENERATION FEMALE RATS 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 - 5 MEAN2S.D. 28.8 5 4.7 [ 181b 29.3 2 9.7 24.0 5 6.1 25.5 2 8.7 23.7 5 5.2 19.4 2 5.8** 17.2 2 7.3** DAYS = DAYS OF LACTATION [ 1 = 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 ( ~ ~ 0 . 0 1 ) . PROTOCOL 418-018: ONE GENERATION REPRODUCTION S T n Y OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 14 (PAGE 2): ABSOLVrE FEED CONSUMPTION VALJUES (G/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE FLATS PREGNANT N 18 18 DELIVERED A LITTER N 18 18 INCLUDED IN ANALYSES N 18 12b FEED CONSUMPTION (G/DAY) DAYS 1 - 5 MEANtS . D . 28.4 5 5.3 20.2 2 5.6** [ Illc DAYS = DAYS OF LACTATION [ 1 = 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 ( ~ ~ 0 . 0 1 ) . 19 19 3b 13.9 2 2.2** , I +I , NI m m d d I +I I w. i/ el dl 0 0 N N P r d d z z + -2 a: \ u W E13 E H ell - 4 B n 2 W 5 cwl! 0 U 8 ?I 3 Wa: d W W W a P a 418-018:PAGE B-50 id I mm I o m m w ,,i d IN d d I , id P P ridd P i . I 0 1 W I ~ wwm d d d 0 h m god 10 m m w id IN d d d P P P d d d 8 H Eiwlii m 4 0 m d * 0 0 0 N N N mmm drid zzz PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 15 (EIAGE 2): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS 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 _t 19.5 65.3 _t 19.1** 46.9 2 6.2** [ lllc _._______.______________________________-----------.---------- DAYS = DAYS OF LACTATION [ 1 = 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 ( ~ ~ 0 . 0 1 ) . I , I m ii I NI 8 , 0 N r P ii ri z z e :101 a w b PJ H- ;i a: 2wa X w ; w n i 418-018:PAGE B-53 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 16 (PAGE I . ) : MATING AND FERTILITY - SUMMARY - Fo GENERATION FEMALE RATS DAYS IN COHABITATION a MEAN2S.D. 2 . 6 2 2 . 4 2 . 7 1.0 3.2 5 2.9 3.0 5 3.0 2.5 5 2.9 3 . 1 5 3.2 2.6 2 2.1 RATS THAT MATED 20(100.0) 19( 9 5 . 0 ) 20 (100.0) 19( 95.0) 27( 96.4) 28(100.0) FERTILITY INDEX b 20/ 20 (100.0) 17/ 19 ( 89.5) 19/ 2 0 ( 95.0) 18/ 19 ( 94.7) 25/ 27 ( 92.6) 27/ 28 ( 96.4) RATS WITH CONFIRMED MATING DATES N 28 20 19 20 19 27 28 MATED BY FIRST MALE c DAYS 1 - 7 N(%) 27( 96.4) 20 (100.0) 19(100.0) 19( 9 5 . 0 ) 19(100.0) 2 6 ( 9 6 . 3 ) 27( 96.4) MATED BY SECOND MALE c DAYS 8 - 1 4 O( 00.0) O ( 00.0) 1( 5 . 0 ) O( 00.0) 1( 3 . 7 ) 1( 3 . 6 ) RATS PREGNANT/RATS IN COHABITATION 20/ 20 (100.0) 17/ 20 ( 85.0) ___-----..__.______^____________________-.--------------------------------------------------.------- 19/ 20 ( 95.0) a. Restricted to rats with a confirmed mating date and rats that did not mate. b. Number of pregnancies/nurnber of rats that mated. c. Restricted to rats with a confirmed mating date. 18/ 2 0 ( 90.0) 418-018:PAGE B-55 N N I 0 * - - +i 0 \ W m W 0 rm N m m rl N 1 N m P N N 2 W $2 V W a a 0 - rl rl 0 - I 0 0 - - +I 0 \ N m 0 0 0 wm N 0 0 0 1 rl N - rl N m N m 0 N - N N m- 0 N* I v W - - - - +I 0 \ w m W m 0 rm N m m dN N m N r d N -- - 9 z - m rp \& z do %'w z z- z I a d d N i) w W rl W E::z E 2 E3 in H J W 0 z 8 W 17 z;J 3 U E:4 0 a: W W J 0 3: U \ a H V 4 d U H 2 0 x 2 in- om hN 14, m km ON W gA $ 9E3 in .. z aw: VZ 3n + OE i3- w L? a: z -in 2g 2 gWPI gz;"2u cn m d 0 m d U' J 0 V 0 b 0 , , , I JI 8 , m N m- 0 NU' - - +I 0 \ W 0 r-m r- d N N m N I c - N d 0 m- NO - - +I 0 0 0 ri N m \o mo Nri N - m N r-- 0 NW - - +I 0 \N 0 mm N d N m r- N 9 - in d*) \z+- 5'w z z - P P 82H u nw x $ 8 z E-, I 5+ 0 Li Eaz3: E; 5 - J H in in 2 2w P h 418-018:PAGE B-56 m- NU' - \ W r-m N m- I NO ~ \o I mo I Nd! -I 8 . IJKO'I'OCOL418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLJE 17 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS TESTED N PREGNANT N(%) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N CORPORA LUTEA MEANAS.D . IMPLANTATIONS MEAN+S.D . LITTER SIZES MEAN+S .D . 1 VEHICLE CONTROL 9 8 ( 88.9) 8 16.1 2 1.6 14.9 2 1.2 13.5 2 1.3 12 1.6 8 8 (100.0) 8 15.4 2 2.7 14.0 5 2.9 13.0 2 2.8 13 2 8 8 (100.0) 8 18.9 2 15.2 5 14.9 2 5.1 1.8 1.7 LIVE FETUSES DEAD FETUSES KESORPTIONS EARLY RESORPTIONS N MEAN+S .D . N MEANAS.D . MEAN+S.D . N MEANkS .D . 108 13.5 2 0 0.0 1.4 2 11 1.4 2 1.3 0.0 1.1 1.1 103 12.9 5 1 0.1 2 1.0 5 7 0.9 2 2.7 0.4 0.9 1.0 119 14.9 2 0 0.0 5 0.4 2 3 0.4 2 1.7 0.0 0.5* 0.5* LATE RESORPTIONS N MEAN+S . D . 0 0.0 A 0 . 0 1 0.1 0.4 0 0.0 2 0.0 b DAMS WITH ANY RESORPTIONS N(%) 8(100.0) 6 ( 75.0) 3( 37.5)** c..L DAMS WITH ALL CONCEPTUSES DEAD OR RESORBED N(%) O ( 0.0) O( 0.0) O( 0.0) DAMS WITH VIABL,E FETUSES N(%) 8 (100.0) 8 (100.0) 8 (100.0) PLACENTAE APPEARED NORMAL N(%) 8(100.0) 8 (100.0) 8 (100. O ) * Significantly different from the Group 1 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value (p50.01.). PKOTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 17 (PAGE 2 ) : CAESAREAN-SECTIONINGOBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 2 1 OF GESTATION N CORPORA LUTEA MEANTS.D . 1M PLANTAT IONS MEANkS . D . LITTER SIZES MEANLS .D . 8 19.1 L 15.4 5 13.6 2 3.3 1.1 2.7 8 17.5 2 4.1 15.5 2 2.4 13.5 2 3.1 8 18.2 2 2.6 16.1 2 1.0 13.6 2 2.4 LIVE FETUSES N MEAN+S . D . 109 13.6 2 2.7 108 13.5 2 3.1 109 1 3 . 6 rfi 2 . 4 DEAD FETUSES N 0 0 0 RESORPTIONS MEAN+S . D . 1 . 8 _t 2 . 2 2.0 2 2.6 2.5 2 3.0 EARLY RESORPTIONS N MEAN2S . D . 14 1.8 L 2.2 15 1.9 2 2.6 19 2.4 2 3.0 LATE RESORPTIONS N MEAN+S . D . 0 0 . 0 _t 0 . 0 J. 0.1 2 0.4 1 0.1 L 0.4 DAMS WITH A" RESORPTIONS N(%) 5 ( 62.5) 6 ( 75.0) 6 ( 75.0) DAMS WITH ALL CONCEPTUSES DEAD OR RESORBED N(%) O( 0.0) O( 0.0) O( 0.0) > DAMS WITH 0 VIABLE FETUSES N(%) 8 (100.0) 8 (100.0) 8 (100.0) M PLACENTAE APPEARED NORMAL N(%) tn v, 8 (100.0) 8(100.0) 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 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 CORPORA LUTEA IMPLANTATIONS LITTER SIZES N MEANLS . D . MEANkS . D . MEANkS,D . 5 CHOLESTEROL CONTROL 8 8 (100.0) 8 17.8 2 2.7 15.9 1: 2.2 15.2 5 2.3 6 1.6 + CHOLESTEROL 8 8 (100.0) 8 17.6 2 14.6 2 14.0 2 3.7 4.2 4.2 7 2 + CHOLESTEROL 8 8(100.0) 8 18.1 2 13.2 2 11.5 2 6.8 4.4 3.9 LIVE FETUSES N MEAN+S .D . 122 15.2 2 2.3 112 14.0 2 4.2 92 11.5 2 3.9 DEAD FETUSES RESORPTIONS N MEANAS.D . 0 0.6 5 0.5 0 0.6 5 1.1 0 1.8 2 1.4 EARLY RESORF'TIONS N MEAN+. D . 5 0.6 2 0.5 5 0.6 2 1.1 14 1.8 L 1.4 LATE RESORPTIONS N 0 0 0 DAMS WITH ANY RESORPTIONS N(%) 5( 62.5) 3( 37.5) 6 ( 75.0) -P c-. DAMS WITH ALL CONCEPTUSES DEAD OR RESORBED N(%) O ( 0.0) O ( 0.0) O ( 0.0) DAMS WITH VIABLE FETUSES N(%) 8 (100.0) 8 (100.0) 8(100.0) m P m r- d m +I m +I +I d N dm m m .4 v d d r m c- in N N m d +I m +I +I 0 0 dm m v N in ri d W N m P d Ti v +I m +I +! 0 m dm m -r m 0 d d r- i d 6' 5' 5' ? ? ? w [il m w w W z z L9 z Ez - :: z[il w W z2 2 q EW iir ki 2 d W iL 5 crj we: 1w E- TnE~Om-. d w hs? n W crj 0 0 i +I rl W d * N m +I 0 dW m ri r W i +I m m W I wi I f/ I di Q. i P N m Q. d * +I w +I 0 in din +I +I in in m W N d d W d i i +I z? in Ti P N m +I 0 dW m d 0 in i +I m 0 r 9 9 n ul 10 10 5`W W z Wz W E E 3: szM wa b kn~E Om-. d z10 ul 4G W E ul 2 $ 3W 2 " 9 W a n z 4 W d ? L4 0 z ii8-4 0 i4 418-018:PAGE B-61 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 18 (PAGE 3): LITTER OBSERVATIONS (CAESAREAN-DELIVEREDFETUSES) - SUMMARY - FO GENERATION FEMALE RATS IMPLANTATIONS MEAN+S . D . 15.9 5 2.2 LIVE FETUSES N MEANkS . D . 122 15.2 2 2.3 POOLED FETAL BODY WEIGHTS (GRAMS)/LITTER MEAN+. D. 7 9 . 4 2 11.8 % DEAD OR RESORBED CONCEPTUSES/LITTER MEAN2S.D. 4.0 2 3.4 * Significantly different from the Group 5 value ( ~ 5 0 . 0 5 ) . 14.6 2 4.2 112 14.0 2 4.2 70.6 2 22.9 4.1 2 7.4 13.2 2 4.4 92 11.5 2 3.9 5 6 . 2 2 18.3 1 2 . 0 2 10.0* PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 19 (PAGE 1): NATURAL DELIVERY OBSERVATIONS - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 1 8 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 MEAN2S.D. 22.9 2 0.3 22.6 5 0.5 9 10 11 12 13 0.8 1 1.2 1.6 2 20 20 20 20 20 17( 85.0) 19( 95.0) 18( 90.0) 17( 85.0) 19( 95.0) 17 (100.O) 19 (100.O) 18 (100. O ) 17 (100. O ) 19 (100. O ) 22.5 2 0.5* 22.4 2 0.6** 22.3 2 0.5** 22.0 2 O . O * * 22.2 A 0.4** IMPLANTATION SITES PER DELIVERED LITTER N MEAN2S.D. 279 232 257 303 275 243 274 14.7 2.3 16.2 1.8 15.1 2.2 15.9 2 2.0 15.3 2 2.5 14.3 2 2.1 14.4 2 1.9 DAMS WITH STILLBORN PUPS N(%) 4( 21.0) 8 ( 40.0)** 2( 11.8) 1( 5.3)* 1( 5.6)* 1( 5.9)* 1( 5.3)* DAMS WITH NO LIVEBORN PUPS N(%) O( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) O ( 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(%) O ( 0.0) O( 0.0) O ( 0.0) 1( 5.3) O ( 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 ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value ( ~ ~ 0 . 0 1 ) . w ?n w PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 19 (PAGE 2 ) : NATURAL DELIVERY OBSERVATIONS - SUMMARY - FO GENERATION FEMALE RATS IMPLANTATION SITES PER DELIVERED LITTER N MEAN+S . D 2C3 14.6 2 2.0 251 13.9 2 2.2 289 15.2 2 1.4 DAMS WITH STILLBORN PUPS N(%) 2 ( 11.1) 4 ( 22.2) 2 ( 10.5) DAMS WITH NO LIVEBORN PUPS N(%) O ( 0.0) O ( 0.0) O ( 0.0) GESTATION INDEX c % N/N 100.0 18/ 18 100.0 18/ 18 100.0 19/ 1 9 DAMS WITH ALL PUPS DYING DAYS 1 - 5 POSTPARTUM N(%) O ( 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 as 0 hour) and the time (in days) the first pup was delivered. c . Number of rats with live offspring/number of pregnant rats. * Significantly different from the Group 2 value ( p . c O . 0 5 ) . * * Significantly different from the Group 2 value (p<O.Ol). PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 19 (PAGE 3) : NATURAL DELIVERY OBSERVATIONS - SUMMARY - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RATS ASSIGNED TO NATURAL DELIVERY N PREGNANT N(%) DELIVERED LITTERS N(%) DURATION OF GESTATION b MEAN2S.D. 5 CHOLESTEROL CONTROL 19a 17( 89.5) 17(100.0) 22.8 2 0.4 6 1.6 + CHOLESTEROL 20 20(100.0) 20(100.0) 22.0 2 0.4** 7 2 + CHOLESTEROL 20 19( 95.0) 19(100.0) 22.3 2 0 . 4 * * IMPLANTATION SITES PER DELIVERED LITTER N MEAN2S.D. DAMS WITH STILLBORN PUPS N(%) 241C 15.1 2 1.4 [ 16lc 2( 11.8) 288 14.4 2 1.7 2( 10.0) 291 15.3 2 2.0 2( 10.5) DAMS WITH NO LIVEBORN PUPS N ( 9 ) O ( 0.0) O ( 0.0) 1( 5.3) GESTATION INDEX d % N/N 100.0 17/ 17 100 I 0 20/ 20 94.7 18/ 19 DAMS WITH ALL PUPS DYING DAYS 1-5 POSTPARTUM N(%) O ( 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 (pc0.01). 418-018:PAGE B-66 w z 0 H b u 4 H E G H b.2 W 0 z El 3 3 W W b.2 U 3 d 0 B in W 4 0 3: U \ n H U 4 U ib.2 x inom hN PI. g &in am ON W S S i-' in e" : zw om uz 3n > 2 8 OEkb? wa: i n z i-n 2% bin wc4 g2 u in S$ a d 0 03 d 6 d 0 U 0 b 0a: 8 , I !Nu) I id . I I dl I I I , , , VI in N N N- W 06 . - . - md P& +I +I m + I 0 0 Nm N 0 m m d Od d din N m m 0 N N- d^ m P 2 P d 4 m +I +I +I d 0 NW m d 6. - MP dN N. - 06 N m m N 11 *c -I - -m. m. NP NP dln rnm Nd \\ WN min d c * - -P. P. inP 6 - -P 6 Nd NN \\ mN do d N N- 0- ^- W 6 $ m m +I +I d in _ - N O 0. - +I 0 0 0 - -.c f M. NW d NU) cf 6N Od d din 06 NN N m m 6 N N- 0- m r 2 md rm- +I +I +I 0 0 Nd in d 0. - mm rim d. - ON \\ wo 6 - -. . cfo do d - -m d mm NN N m 6 m N N- 0- N m P r +I +I d 6 +I 0 0 NLI) 6 d 6. - 6m d6 d. - ON \\ *a N - -6. W. OW - -m c f 66 NN N \\ m 6 W N N- 0- O 0 . -g . - 0 0 + I + I N 0 mo W +I m 0 6 in 6 d o m d dm dW d - -0. 6. dd - -d m ma NN N W P W \\ m6 - - N N- 0- d m cfm 0 mw g -. -. . - . - m m +I +I +I d 0 ON 'N r\ d W mm ~m P m n om v) N m m o om W WVI d dW N NN N H \\ - - 9 - . - ? a 5% B z in in dQ i n * ; 1$ s1W x W I: W I: in 5 I E 24 0 b - n W a W 3 F 2 3 4 W 0 0 m 2 n m d W 4 8 in H 2 i4 3 d 2 i4 4 dW 4 * z U do 1- z z a \ z \ z W in W in 3 2 e: $ 0 2 d Wn m 2 dN 5 5 in > >in i4 E u 3 i-1 u a m 0 a d x a c m a 4 E0 ' Ra,d >o .il I do 7 3V I 2 > h a , a,? Rd Em \d E? a 0s i-10 am i -u1 JJ ma, 0 a us inE hh am w 4 18-018:PAGE B-67 E z z :2 : E m H 4 W 0 2 8ziw r3. 4 4 s U 4 0 a: W $ W 4 0 3: U \ 2 2 2zU H 0 4 z m- om hN a. m cr,m ON W gA t; .. z nw: om uz 4z 3n > O2 Eh? wo 3 zz - z8 zw;"a m rn Wz aE: on: m-- 0 m d * 4 0 s;i 0 n: 14 I , m d r ri m d I rit I m d r d , 0 N m ri z o * * +* *+ *I m. *. m . * . * . N**** +I +I +I +I +I N. * r. n - . a " . m . M m N N N d *** +I +I +I +I +I *. O. d . W . W . mmlcww d m. N . m . * .i n . ~ m m m +I +I +I +I +I o. m . r . i w. q. *NNrid dridrid m. o. o. 0. 0. am*** +I +I +I +I +I ommmm NNriNN +I +I +I +I +I v. m. ~. m. .- vmmmm riridriri *. *. *. *. *. N N N N N +I +I +I +I +I * * * * * W. * . m . " . . ririridd r . - . m .m v. - . N N N N N +! +I +I +I +I . . . . . P i n * * r l mmmmm riddriri ..... nnaan 0 *ri +I ri 0 m 10 m +I m *in W * d W * * m ri ii +I +I 0 W 0 in m * m 0 m P ri ri +I +I m m W * m* W * W * W * m W 0 d P ri ri d ri 0 d ri ri ri d +I +I +I +I +I ; ~ m 0 m r- m m * * * * m m m m m d m * m ri ri +I +I Cr 0 W * W * * ri r r ri d +I +i 0 m 0 0 in m Pm m m m m ii d ri +I +I +I N d ri W * W * W -9 0 0 m P P W ri ri d +I +I +I *W P 0 0 0 m in m S' S' SI 5' 9 9 9 9 9 o 10 rn 10 rn W z w r W z W z s N m >a: n > 2 2 N m -s in 2 n > 2 > 2 2 a 2>i PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 20 (PAGE 3 ) : LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS ________________________________________-------------------------------------------------------------------------------------------- DOSAGE GROUP 1 8 9 10 11 12 13 DOSAGE MG/KG PFOS VEHICLE CONTROL 0 . 4 0.8 1 1.2 1.6 , 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) MEANfS . D . MEANfS . D . MEANfS .D . MEANfS .D . MEAN2S.D . MEANkS .D . 86.9 2 15.4 85.2 2 15.1 84.2 94.4 f 15.1 92.0 f 14.7 87.8 103.8 2 16.1 99.5 2 17.4 94.0 116.6 f 18.2 109.5 f 19.7 104.2 [ 18lb 129.4 f 19.8 121.5 f 21.2 112.6 +42.5 f 12.4 +36.3 2 12.7 +28.5 f 10.4 85.8 f 14.3 77.2 f 1 2 . 1 84.2 2 20.2 74.5 f 13.8 f 17.9 2 20.8 f 17.5 95.1 % 15.7 76.0 [ 18la 105.1 2 17.6 79.2 [ 18la 113.0 2 19.1 85.2 [ 18la t25.9 2 11.8**+7.9 [ 18la f 16.2 f 20.2** f 25.4** f 28.2** f 32.2** f 26.5** 67.0 2 15.9** 52.8 f 26.0** [ 14la 51.5 f 30.5** [ 14la 55.0 f 34.8** [ 14la 63.5 f 37.4** I 13la -3.2 f 33.0** [ 13la 58.5 f t 18la 33.5 f t llla 44.4 f [ 7la 54.1 2 [ 61a 68.3 2 t 51a -9.1 2 [ 5la 25.6** 28.7** 36.3** 38.7** 38.8** 35.7* AVERAGE PUP WEIGHT (GRAMS) DAY 1 MEAN2S.D . 6.4 f 0.4 5.9 f 0.6* 5.9 f 0.6* 5.8 2 0.8** 5.7 f 0.6** 5.4 f 0.5** 5.3 f 0.5** DAY 2 DAY 3 MEAN2S.D . MEAN2S.D . 7.1 2 0.6 7.8 2 0.7 6.4 f 0.5* 7.0 f 0.7** 6.4 f 0.8** 6.1 f 0.8** 6 . 9 f 1.0** 6 . 7 f 0 . 9 * * 5.8 f 0.7** 6 . 2 f 1.0** 5.4 f 0.5** [ 141a 5.8 f 0.8** [ 181a 5.2 f 0.8** [ Illa 5 . 9 2 1.1** DAY 4 MEAN2S.D . 8.8 f 0.9 7.8 f 0.9* 7.8 2 1.2* [ 181a 7 . 5 f 1.1** 6 . 8 f 1.1** [ 14la 6 . 3 f 1.0** [ 71a 6.5 f 1.4** DAY 5 [ 18lb t 1813 [ 14la [ 61a MEANfS . D . 9 . 8 2 1.1 8 . 6 2 1.0** 8 . 5 2 1 . 4 * * 8 . 1 2 1 . 3 * * 7 . 4 f 1 . 4 * * 7 . 1 f 1.1** 7 . 4 f 1 . 5 * * P c DAYS 1 - 5 . . [ 18la [ 131a [ 5la 00 MEANfS D + 3 . 4 f 0 . 9 + 2 . 7 f 0 . 7 * + 2 . 5 f 0 . 9 * + 2 . 2 f 0 . 8 * * + 1 . 7 2 1 . 2 * * + 1 . 7 f 1.0** + 1 . 5 2 1 . 2 * * (BODY WEIGHT CHANGE) ___________________________________ [ 18la [ 13la t 5la DAY = DAY POSTPARTUM [ I = 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 ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 1 value (p.cO.01). PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATlON IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 20 (PAGE 4): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS _.-__________.___.______________________- DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 18 18 19 PUPS DELIVERED (TOTAL) N MEAN+S . D 237 13.2 2 2.2 233 12.9 2 2.3 270 14.2 2 1.4 LIVEBORN STILLBORN 13.0 2 2.2 234( 98.7) 0.2 2 0.5 3( 1 . 3 ) 12.6 2 2.1 227( 97.4) 0.3 2 0.7 6( 2.6) 14.1 A 1.4 268( 99.2) 0.1 + 0.3 2( 0.7) UNKNOWN VITAL STATUS N 0 0 0 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2- 5 1/234( 0.4) 2/233( 0.8) 22/227( 9.7) 111/205 ( 54.1) 93/268( 34.7)* * 172/175( 98.3)** VIABILITY INDEX a % 98.7 41.4 1.1 N/N 231/234 94/227* 3/268** . _ . _ _ . . _ _ _ _ _ _ _ _ _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _. _ . . . ._ . . .~ . . .~ . . . ~ . . . .~ . . . ~ . . . .~ ~ ~ ~ ~ ~ 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 2 value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 2 value ( ~ ~ 0 . 0 1 ) . PROTOCOL 41.8-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 0 (PAGE 5 ) : LITTER OBSERVATIONS (NATURALLYDELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS SURVIVING PUPS/LITTER a DAY lb MEAN+S .D . DAY 2 MEAN+S . D . DAY 3 DAY 4 DAY 5 MEAN+S . D . MEAN+S .D . MEAN2S . D . 13.0 + 2.2 12.9 5 2.2 12.8 2 2.3 12.8 2 2.3 12.8 2 2.3 12.6 2 2.1 6.2 2 4.6** 5.6 2 4.8** 5.2 2 4.8** 5.2 2 4.8** 14.1 2 1.4 1.4 2 2.0** 0 . 5 2 1.1** 0.3 2 0.2 + 0.8** 0.5** PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY lb MEAN2S.D . 56.2 + 12.4 49.9 2 10.8 48.0 2 15.6 DAY 2 MEAN+S .D . 56.4 2 12.2 55.0 2 18.0 54.6 2 41.1 DAY 3 MEAN+S .D. 55.8 5 12.3 [ 15lc 49.8 2 12.4 [ SIC 23.3 2 32.5** DAY 4 MEAN+S .D . 55.8 2 12.3 [ 13lc 52.2 2 10.4 [ 51c 55.6 2 50.9 DAY 5 MEAN+S .D . 55.8 2 12.3 [ 12lc 52.2 5 10.4 [ 31c 50.0 2 70.7 [ 12lc [ 2lc .________.._____________________________---.---------------.-- DAY = DAY POSTPARTUM [ 1 = 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 ( ~ ~ 0 . 0 1 ) . PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 20 (PAGE 6 ) : LITTER OBSERVATIONS (NATURALLYDELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS POOLED PUP WEIGHTS/LITTER (GRAMS) DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAYS 1 - 5 (BODY WEIGHT CHANGE) MEANfS .D . MEANAS.D . MEANfS .D . MEANAS.D . MEANkS . D . MEANfS . D . 83.0 f 11.5 90.6 f 11.6 99.7 f 14.4 112.1 5 15.5 123.9 2 17.1 +40.9 2 8.2 63.1 2 18.7** 43.6 f 25.9** [ 15la 50.1 A 28.6** [ 13la 60.6 2 35.9** [ 12la 63.8 f 33.4** [ 121a -5.6 f 32.5** [ 12la 57.1 f 22.2** [ 16la 17.5 f 9.8** [ 8la,b 11.0 5 9.1** [ 5la 12.1 f 7.1** [ 3la 11.0 A 8.9** [ 2la -39.7 5 33.5 [ 21a AVERAGE PUP WEIGHT (GRAMS) DAY 1 MEANAS.D . 6.4 f 0.5 5.6 f 0.5** DAY 2 MEANfS .D . 7.1 f 0.6 5.6 _i 0 . 8 * * DAY 3 MEANkS .D . 7.8 f 0.7 [ 15la 6.1 f 1.2** DAY 4 MEANfS . D . 8.8 k 0.8 I 13la 7.2 k 1.7** . . [ 12la DAY 5 MEANfS D 9 . 8 k 1.0 7.7 f 1 . 8 * * -P c.' [ 12la 00 DAYS 1 - 5 MEANfS . D . +3.3 f 0 . 8 +2.0 % 1.4** I 0 (BODY WEIGHT CHANGE) [ 12la w ________________________________________-------------------------------------- PP DAY = DAY POSTPARTUM [ 1 = 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 (~50.05). * * Significantly different from the Group 2 value ( ~ ~ 0 . 0 1 ) . PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 0 (PAGE 7 ) : LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS DOSAGE GROUP DOSAGE MG/KG PFOS -_________.._______.____________________. DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 17 20 18 PUPS DELIVERED (TOTAL) LIVEBORN N MEAN+S .D MEAN+S .D . N(%) STILLBOJLN 247 14.5 2 2.2 14.4 2 2.2 245( 99.2) 0.1 2 0.3 2 ( 0.8) 271 13.6 + 2.1 13.4 2 2.3 269( 99.3) 0.1 5 0.3 2 ( 0.7) 261 14.5 5 1.7 14.4 5 1.8 259( 99.2) 0.1 2 0.3 2 ( 0.8) UNKNOWN VITAL STATUS N 0 0 1 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2 - 5 0/245( 5/245( 0.0) 2.0) 18/269 ( 6.7) * 138/251( 55.0) * 27/258 ( 10.5)** 194/231( 84.0) ** VIABILITY INDEX a % 98.0 42.0 14.3 N/N 240/245 113/269* 37/258** _____.__________________________________------.------------. 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 ( ~ ~ 0 . 0 5 ) . * * Significantly different from the Group 5 value ( ~ ~ 0 . 0 1 ) . PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 20 (PAGE 8): LITTER OBSERVATIONS (NATURALLYDELIVERED PUPS) - SUMMARY - F1 GENEFATION LITTERS 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 . 14.4 k 2.2 14.3 2.2 14.2 k 2.1 14.2 2 2.1 14.1 5 2.0 13.4 2 2.3 8.4 + 4.6** 6.0 2 5.5** 5.8 2 5.4** 5.6 2 5.5** PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY 1b MEAN+S .D . 48.3 2 14.5 47.3 + 16.8 DAY 2 MEAN+S . D . DAY 3 MEANAS.D . DAY 4 MEAN+S . D . DAY 5 MEANAS.D . .________ 48.2 2 14.9 48.2 2 14.8 48.2 + 14.8 48.4 5 14.9 50.0 5 24.0 [ 19lc 41.1 + 26.8 [ 14lc 44.4 5 25.0 [ 13lc 43.9 2 26.0 [ 12lc DAY = DAY POSTPARTUM [ 1 = 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 ( ~ ~ 0 . 0 1 ) . 14.4 5 1.8 4.0 5 2.2 + 4.4** 4.0** 2.2 5 3.9** 2.0 2 4.0** 56.1 2 11.9 59.8 2 32.2 [ 14lc 49.6 2 33.9 [ 71c 48.6 + 33.8 [ 71 c 48.0 5 21.4 [ 51c ------------ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE ANE NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 20 (PAGE 9): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - F1 GENERATION LITTERS POOLED PUP WEIGHTS/LITTER (GRAMS) DAY 1 MEANkS .D . DAY 2 MEANkS.D . DAY 3 DAY 4 DAY 5 MEAN+S .D . MEAN+S .D . MEAN+S .D . DAYS 1 - 5 (BODY WEIGHT CHANGE) MEAN2S.D . 90.9 2 12.5 100.7 5 18.9 108.7 2 16.8 123.4 2 17.7 134.4 5 20.3 [ 16lb +44.4 5 11.8 [ 16lb 67.7 2 14.1** 47.8 5 25.5** [ 19la 53.2 2 30.6** [ 14la 62.5 2 32.4** [ 1313 73.1 5 32.0** [ 12la +3.3 5 31.1** [ 12la AVERAGE PUP WEIGHT (GRAMS) DAY 1 MEAN2S.D . 6.3 2 0.5 5.4 2 0 . 4 * * DAY 2 MEANkS .D . 7.1 2 0.8 5.2 2 0.6** DAY 3 MEANkS .D . 7.7 2 0.8 [ 19la 5 . 8 2 0.9** DAY 4 MEAN+S . D . 8.8 2 0.9 [ 14la 6.7 2 l.l** DAY 5 MEAN2S.D . 9.6 5 1.1 [ 13la 7.5 2 1.1** DAYS 1 - 5 MEANkS .D . [ 16lb +3.3 5 0.7 [ 12la +2.0 2 0.9** (BODY WEIGHT CHANGE) 16lb [ 12la ________________________________________-------------------------------------- DAY = DAY POSTPARTUM [ 1 = 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 ( ~ ~ 0 . 0 1 ) . 68.6 2 20.2** 26.7 5 22.8** [ 14la 32.6 2 27.1** [ 71a 33.9 2 29.3** [ 71a 47.9 5 26.2** [ 51a -24.3 5 24.1** [ 51a 5.3 2 0.4** PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 21 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - SUMMARY - F 1 GENERATION PUPS ._____.......______.____________________... MATERNAL DOSAGE GROUP 1 9 10 11 12 13 DOSAGE MG/KG PFOS VEHICLE CONTROL 0.8 1 1.2 1.6 2 L,ITTERS EXAMINED (N) 19 17 19 18 17 19 . _ . . - - - - . . _ _ _ _ _ . _ . _ _ _ _ _ _ _. _ ._ . ._ - - _-_ . - _ _ _ _ __ __ __ __ __ __ __ __ - ._ ~. - - - - - - - - - - - - - - - - - - - - - - - - - . - TRANSIENT CLINICAL OBSERVATIONS: a TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS COLD TO TOUCH N/N 1/1 NOT NURSING N/N o/o DECREASED MOTOR ACTIVITY N/N o/o GASPING N/N o/o LABORED BREATHING N/N o/o NOT NESTING N/N o/o PERSISTENT CLINICAL OBSERVATIONS: a TAIL MISSING FROM BASE N/N o/o TIP OF TAIL MISSING N/N o/o TIP OF TAIL BLACK N/N 2/1 .... ____._____.___________________________ 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 I w , ri8 H 2:: mE- H 5 4 W 0 2 m CL 3 a W 8 17 H 3 d 2 d 2u WzW r.9 4 0 ac d is W E- m W J 5 0 zu E \ m E:u A 5 U H 2 8 pi J 2 E z 0 a in m - $t om d i s m pi. in 0 i r , m E- ON W 3: gE! 2 F S H B Gl .. m z dw z 0 om d ir, u z G3 8 3n > on H b g g 9 w m a: z - Wm oac m HO 0 bm 4 $8 4 wa U z;? zii 0 w c! r WE- U E $ -N Q d 0 W u Q 2 ri dl 1 2 iiw J ri 0 N U 0 u: b , in d( 0 0 d \ \ \ \ \ m N 0 0 rl m 4 m ri d d \ \ \ \ m m ri rf d 0 0 0 0 \ \ \ \ 0 0 0 0 E z z 2 z id , .. \ z \ z \ z \ z m 2 2 > 2i E2 m G z m 0 A u 2 ac J 5 0 0 3: E- E- H E r.9 V 3 0 z 10 0 ? w E E- a J u 0 Wm x b 3 3 z E- 4 H G b a 4 auc z 0 z 0u Wn 0 z H m e: W a 418-018:PAGE B-76 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 21 (PAGE 3): CLlINICAL OBSERVATIONS FROM BIRTH TO DAY 5 POSTPARTUM - SUMMARY - F1 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 o/o 6/3 49/8** 1/1 49/5 27/6 o/o o/o 4/2 o/o 2/1 13/2 4/2 o/o o/o 1/1 o/o o/o E'ERSISTENT CLINICAL OBSERVATIONS: a DISTAL PORTION OF TAIL MISSING TIP OF TAIL BLACK N/N 3/1 o/o o/o TIP OF TAIL MISSING N/N 2/1 o/o o/o ___.__..___.________.~~~~~~~~~~~~~~~.~~~~~~~~ so STATISTICAL ANALYSES WERE RESTRICTED TO THE NUMBER OF LITTERS WITH OBSERVATIONS. N/N = TOTAL FREQUENCY (DAYS x PUPS)/LITTERS WITH OBSERVATIONS P L a. Tabulation restricted to adverse observations; all other pups appeared normal. b. Located on the right forelimb and right forepaw. 0 * * Significantly different from the Group 5 value (p~O.01). c- PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 22 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - F1 GENERATION PUPS MATERNAL DOSAGE GROUP 1 8 9 10 11 12 13 DOSAGE MG/KG PFOS VEHICLE CONTROL 0.4 0.8 1 1.2 1.6 2 LITTERS EXAMINED N 19 20 17 19 18 17 19 TOTAL UNSCIIEDULED NECROPSIES a N 5 12 STILLBORN N 4 8 FOUND DEAD N 1 4 8 13 17 43 91 2 2 1 4 3 6 11 16 39 88 NO MILK IN STOMACH b N(%) l(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) O( O.O)** O ( O.O)** O ( O.O)** O ( O.O)** O ( O.O)** O ( O.O)** ..___________.___.______________________~..~~----~~-.-~~~ SCHEDULED PUP NECROPSIES ON DAY 5 POSTPARTUM LITTERS EVALUATED N 19 20 17 18 18 13 5 PUPS EVALUATED N 253 274 228 253 206 112 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. O ) 253(100.0) 18 (1.00 . O ) 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) 1 ( 5.0) O ( 0.0) O( 0.0) O ( 0.0) O( 0.0) O ( 0.0) PUP INCIDENCE N(%) O ( 0.0) 1( 0.4) O( 0.0) O( 0.0) O ( 0.0) O( 0.0) O ( 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 ~ O . 0 1 ) . Complete necropsies were not performed on pups in which w z 0 H 8E- m - r - N L o Lo 0 1 Nm H Loin Lo d d d -/ z W W 01 0 m F d 0 a 10 W d 0 z U 2 \ a H U a: m a d iii m z 0 H 8 > m a 0 w z I N A N 5 3 u0 N 0 W ti 0 ti - a owwm -/ 0 1 N* dm 0 0 md m rim N zzz zz a a w w 0 z 418-018:PAGE B-79 , 8 , tc i N mmrl wm mw m Nm drl rl in P. 3 CL 5 8 H W z; 17 d h z 9 m Lzl 0 5 H t. a: m W m 0 z > in P. 0 up: W m W 17 -P4I N N !iW F. 418-018:PAGE B-80 PROTOCOL,418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN m r s (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 VEHICLE CONTROL RAT # DESCRIPTION 10901 INCISORS: MISSING/BROKEN a 10902 RED PERIVAGINAL SUBSTANCE 10903 NO ADVERSE FINDINGS 10904 DG( 0- 5) INCISORS: MISSING/BROKEN 10905 NO ADVERSE FINDINGS 10906 NO ADVERSE FINDINGS 10907 NO ADVERSE FINDINGS 10908 NO ADVERSE FINDINGS 10909 DG( 12 ) INCISORS: MISSING/BROKEN 10910 NO ADVERSE FINDINGS 10911 NO ADVERSE FINDINGS 10912 NO ADVERSE FINDINGS 10913 NO ADVERSE FINDINGS 10914 DL( 2 - 5 ) INCISORS: MISSING/BROKEN a DL( 3 ) INCISORS: MISALIGNED 11501 DS( 32.- 43) LOCALIZED ALOPECIA: UNDERSIDE DG( 0- 2 ) LOCALIZED ALOPECIA: UNDERSIDE DG( 5- 8 ) LOCALIZED ALOPECIA: NECK DG( 16- 21) LOCALIZED ALOPECIA: UNDERSIDE DL( 1- 5) LOCALIZED ALOPECIA: UNDERSIDE a 11502 DS( 9- 18) LOCALIZED ALOPECIA: LIMBS DS( 28- 34) LOCALIZED ALOPECIA: LIMBS 11503 NO ADVERSE FINDINGS 11504 NO ADVERSE FINDINGS 11505 NO ADVERSE FINDINGS 11506 NO ADVERSE FINDINGS 90 11507 NO ADVERSE FINDINGS P 11508 NO ADVERSE FINDINGS c-. 11509 DS( 27- 39) RIGHT FOREPAW: 2ND DIGIT SWOLLEN DS( 27- 42) RIGHT FOREPAW: 2ND DIGIT MISSING 0 DG( 0- 21) RIGHT FOREPAW: 2ND DIGIT MISSING DL%( 1- 5 ) RIGHT FOREPAW: 2ND DIGIT MISSING a DL( 2 ) RED PERIVAGINAL SUBSTANCE 11510 DS( 5- 14) LOCALIZED ALOPECIA: LIMBS DS( 23- 42) LOCALIZED ALOPECIA: LIMBS DG( 0- 21) LOCALIZED ALOPECIA: 1,IMBS DL( 1- 5) LOCALIZED ALOPECIA: LIMBS a DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 1 VEHICLE CONTROL RAT # DESCRIPTION 11511 NO ADVERSE FINDINGS 11512 NO ADVERSE FINDINGS 11513 DS( 3- 42) TAIL BENT DS( 40- 42) LOCALIZED ALOPECIA: UNDERSIDE DG( 0- 21) TAIL BENT DG( 0 - 21) LOCALIZED ALOPECIA: UNDERSIDE DL( 1- 5) TAIL BENT a DL( I- 5) LOCALIZED ALOPECIA: UNDERSIDE a 11514 NO ADVERSE FINDINGS ...._.._________._______________________~~~~~.~~~~ DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 3): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10915 10916 10917 10918 10919 10920 10921 10922 10923 10924 10925 10926 10927 10928 DG( DG( DG( DG( DG( DS( DS( DS( DG( DG( DG( DG( DG( DG( DG( DG( DG( DG( DG( DG( DS( DS( 1 ) 4- 9) 13- 21) 10- 12) 17 ) 10 ) 34 ) 41- 45) 0- 2) 4- 5) 10 ) 13 1 15 ) 17 ) 17 ) 19 17 ) 19 ) 15 ) 18 ) 37- 40) 43- 44) NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: UNDERSIDE a MOUTH: SCAB (0.1 CM IN DIAMETER) CHROMORHINORRHEA EXCESS SALIVATION EXCESS SALIVATION RIGHT EAR SWOLLEN RIGHT EAR SWOLLEN EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION RED, DRIED PERIORAL SUBSTANCE EXCESS SALIVATION MODERATE PERIORAL SUBSTANCE RED, MODERATE PERIORAL SUBSTANCE EXCESS SALIVATION CHROMORHINORRHEA RED PERIVAGINAL SUBSTANCE RED, SLIGHT PERIORAL SUBSTANCE INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN EXCESS SALIVATION EXCESS SALIVATION EXCESS SALIVATION NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA EXCESS SALIVATION SOFT OR LIQUID FECES DECREASED MOTOR ACTIVITY CHROMORHINORRHEA a CHROMODACRYORRHEA a RED PERIORAL SUBSTANCE a MORIBUND SACRIFICED; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN ----_---__-.-__--_______________________----.------ DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PKO?'OCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS . . _ _ _ _ _ _ . . __ _ __ _._ . _ _ _ _ . _ _ _ . . . . . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ~ ~ . - ~ ~ ~ ~ DOSAGE GROUP 2 MEVALONIC ACID CONTROL _._.____________._..____________________~~~..-~~-----.~~ RAT # DESCRIPTION ..____.._______..___.~..~~~~~~~~~~~~~..~~~~~~~~.-.--~ 11515 DG( 16 ) EXCESS SALIVATION 11516 NO ADVERSE FINDINGS 11517 DS( 3 3 - 35) INCISORS: MISSING/BROKEN 11518 DG( 17 ) EXCESS SALIVATION 11519 DS( 6 ) RALES DS( 13 ) CHROMORHINORRHEA 11520 DS( 28- 32) LOCALIZED ALOPECIA: UNDERSIDE DS( 40- 44) LOCALIZED ALOPECIA: UNDERSIDE DG( 0- 2) LOCALIZED ALOPECIA: UNDERSIDE DG( 1 4 - 17) LOCALIZED ALOPECIA: UNDERSIDE DG( 15 ) INCISORS: MISSING/BROKEN 11521 NO ADVERSE FINDINGS 11522 DG( 17 ) RED PERIORAL SUBSTANCE 11523 DG( 11 ) EXCESS SALIVATION DG( 18 ) EXCESS SALIVATION DG( 18 ) RED, SLIGHT, DRIED PERIORAL SUBSTANCE 11524 NO ADVERSE FINDINGS 11525 DG( 12- 21) LOCALIZED ALOPECIA: LIMBS DL( 1- 5) LOCALIZED ALOPECIA: LIMBS a 11526 DG( 20 ) EXCESS SALIVATION 11527 NO ADVERSE FINDINGS 11528 DS( 6 ) EXCESS SALIVATION ___.________.._____..~~~~~.~~~-~~~~.~~~~~~~~---~~----- DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION 0 M PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 5): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10929 NO ADVERSE FINDINGS 10930 DS( 40- 42) SWOLLEN SNOUT DS( 40- 44) MOUTH: SCAB (0.1CM IN DIAMETER) DG( 0 - 3) MOUTH: SCAB (0.1 CM IN DIAMETER) DG( 7 ) CHROMORHINORRHEA DG( 7 SWOLLEN SNOUT DG( 9 ) EXCESS SALIVATION DG( 13 ) EXCESS SALIVATION DG( 15 ) EXCESS SALIVATION 10931 NO ADVERSE FINDINGS 10932 NO ADVERSE FINDINGS 10933 LOCALIZED ALOPECIA: NECK 10934 EXCESS SALIVATION 10935 EXCESS SALIVATION EXCESS SALIVATION 10936 FOUND DEAD 10937 NO ADVERSE FINDINGS 10938 EXCESS SALIVATION 10939 NO ADVERSE FINDINGS 10940 DG( 19 ) RED, SLIGHT PERIORAL SUBSTANCE DG( 19- 21) INCISORS: MISSING/BROKEN a 10941 DL( 4 ) SACRIFICED DUE TO NO SURVIVING PUPS 10942 DG( 13- 21) LOCALIZED ALOPECIA: LIMBS DG( 18- 21) LOCALIZED ALOPECIA: UNDERSIDE DLj( 1 - 5 ) LOCALIZED ALOPECIA: LIMBS a DL( 1- 5) LOCALIZED ALOPECIA: UNDERSIDE a 11529 DS( 6 ) DS( 33- 35) EXCESS SALIVATION INCISORS: MISSING/BROKEN 11530 NO ADVERSE FINDINGS 11531 LOCALIZED ALOPECIA: UNDERSIDE 11532 SACRIFICED DUE TO NO SURVIVING PUPS 11533 EXCESS SALIVATION CHROMORHINORRHEA SACRIFICED DUE TO NO SURVIVING PUPS .._____---__________..-.-.------.-------.--------.------------------------ DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABIJE 23 (PAGE G ) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ____._.._________._.____________________..~~~~~~.~~~ DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID ..._.....____.._________________________~~~~~~~~~~~~~.~ RAT # DESCRIPTION 11534 11535 11536 11537 11538 1153Y 11540 11541 11542 11543 DL( 2 ) DS( 28- 32) DS( DS( DG( DS( DS( DS( DS( DS( DS( DS( DL( 14- 18) 19- 23) 12 ) 4 ) 4 ) 4 - 6) 8- 12) 12 ) 12 ) 12 ) 3 ) DL( 2 ) DG( 13 ) DG( 16 ) DS( 41- 44) DG( 0 ) DG( 15 ) NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING PUPS INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN MOUTH: SCAB (0.1CM 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 P c 00 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 7): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ...__..._......_ ..._ ... .......................................................................................................... DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID . _ . _ _ .. . _ . _ _ _._ _ . _ _ _ _ _ _ _ _ _ _ _ _ . . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ . ~ ~ ~ ~ ~ ~ ~ ~ . ~ RAT li DESCRIPTION _ _ _ _ _ . _ _ . _ _ _ . . . . _ _ __ _ . _ __ __ __ __ _ __ __ __ __ _ _ __ . _ __ _. __ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - 10943 DL( 3 ) SACRIFICED DUE TO NO SURVIVING PUPS 10944 DI,( 5 ) SACRIFICED DUE TO NO SURVIVING PUPS 10945 DS( 32 ) EXCESS SALIVATION DS( 34- 35) EXCESS SALIVATION DS( 38- 39) EXCESS SALIVATION DS( 41 EXCESS SALIVATION DG( 0- 1) EXCESS SALIVATION 10946 NO ADVERSE FINDINGS 10947 DG( 17- 21) LOCALIZED ALOPECIA: LIMBS a 10948 DS( 34 ) EXCESS SALIVATION DL( 4 ) SACRIFICED DUE TO NO SURVIVING PUPS 10949 DS( 4 - 44) TIP OF TAIL MISSING DS( 37 ) CHROMORHINORRHEA DG( 0- 21) TIP OF TAIL MISSING a DG( 14- 15) CHROMODACRYORRHEA DG( 14- 16) CHROMORHINORRHEA DG( 14- 18) RALES DG( 14- 21) PTOSIS DG( 15- 18) RED, SLIGHT OR MODERATE PERIORAL SUBSTANCE DG( 15- 19) DEHYDRATION DG( 17 ) SCANT FECES DG( 17- 18) EMACIATION 10950 NO ADVERSE FINDINGS 10951 DS( 38 ) CHROMORHINORRHEA DG( 14 ) EXCESS SALIVATION 10952 DG( 8 ) EXCESS SALIVATION DG( 16 ) EXCESS SALIVATION DL( 2 ) SACRIFICED DUE TO NO SURVIVING PUPS 10953 DS( 34 ) EXCESS SALIVATION DG( 20- 21) INCISORS: MISSINGjBROKEN a 10954 DS( 12 ) SOFT OR LIQUID FECES DS( 14 ) CHROMORHINORRHEA DS( 15- 17) INCISORS: MISSINGjBROKEN DS( 16 ) SOFT OR LIQUID FECES DG( 21 ) INCISORS: MISSING/BROKEN a -.----__..-___--___-____________________.-----------.---------------- DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 8): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 10955 EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS 10956 SACRIFICED DUE TO NO SURVIVING PUPS 11544 SACRIFICED DUE TO NO SURVIVING PUPS 11545 EXCESS SALIVATION SWOLLEN S N O W MOUTH: SCAB (0.2 CM IN DIAMETER) INCISORS: MISALIGNED INCISORS: MISSING/BROKEN SACRIFICED DUE TO NO SURVIVING PUPS 11546 SACRIFICED DUE TO NO SURVIVING PUPS 11547 EXCESS SALIVATION RED, SLIGHT PERIORAL SUBSTANCE EXCESS SALIVATION EXCESS SALIVATION 11548 NO ADVERSE FINDINGS 11549 RED, SLIGHT, DRIED PERIORAL SUBSTANCE SACRIFICED DUE TO NO SURVIVING PUPS 11550 EXCESS SALIVATION SACRIFICED DUE TO NO SURVIVING PUPS 11551 RED, SLIGHT PERIORAL SUBSTANCE CHROMORHINORRHEA CHROMORHINOFtRHEA SACRIFICED DUE TO NO SURVIVING PUPS 11552 EXCESS SALIVATION PTOSIS PTOSIS EXCESS SALIVATION RED, MODERATE PERIORAL SUBSTANCE PTOSIS 11553 SACRIFICED DUE TO NO SURVIVING PUPS 11554 EXCESS SALIVATION RED, SLIGHT PERIORAL SUBSTANCE SACRIFICED DUE TO NO SURVIVING PUPS 11555 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 PROTOCOL 418-018: 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 E'ROTOCOL~ 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 10): CLINICAL OBSERVATIONS - INDIVIDTJAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 5 CHOLESTEROL CONTROL ._._.___.____..____.____________________~~~~.~~~~~~~.~ RAT # DESCRIPTION _._____.._..____.___~.~.~~~.~..~~~~..~~-~~~~~~~~~.~~~~~ 1.0957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 DG( 6 - 15) DG( 19- 21) DS( 35- 44) DS( 37- 44) DG( 0- 6) DG( 0- 21) DG( 11- 21) DS( 39 ) DG( 9- 20) DL( 1- 5) DL( 4- 5) DS( 22- 32) DS( 16 ) DS( 16 ) DS( 16 ) DS( 16 ) DL( 2 ) DS( 23- 32) DG( 19- 21) DL( 1- 5) DS( 27- 45) DG( 0- 25) INCISORS: MISALIGNED NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMODACRYORRHEA a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: NECK LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: NECK a LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a LOCALIZED ALOPECIA: LIMBS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS NO ADVERSE FINDINGS DECREASED MOTOR ACTIVITY PALE IN APPEARANCE COLD TO TOUCH MORIBUND SACRIFICED RED PERIVAGINAL SUBSTANCE LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 2 3 (PAGE 11): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 5 CHOLESTEROL CONTROL RAT # DESCRIPTION ____...__________._.____________________~~~~~~~~~. 11568 DS( 25 ) SWOLLEN SNOUT DS( 25- 40) INCISORS: MISALIGNED DS( 42- 44) INCISORS: MISALIGNED DG( 0- 21) INCISORS: MISALIGNED DL( 1- 4) INCISORS: MISALIGNED 11569 DL( 2 ) RED PERIVAGINAL SUBSTANCE 11570 DS( 40- 43) LOCALIZED ALOPECIA: NECK DS( 40- 45) LOCALIZED ALOPECIA: UNDERSIDE DG( 0- 1) LOCALIZED ALOPECIA: UNDERSIDE DG( 10- 21) LOCALIZED ALOPECIA: UNDERSIDE DL( 1- 4) LOCALIZED ALOPECIA: UNDERSIDE 11571 NO ADVERSE FINDINGS ____._________._________________________------------------------------------------.---- US IDAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION cp 0 PKO'I'OCOI, 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 12): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 6 RAT # 10971 10972 10973 10974 10975 10976 10977 10978 10979 1.0980 10981 DL( 3 ) DS( DG( DG( DG( DL( DI,( DL( DS( 35- 42) 0- 2) 14- 19) 14- 20) 1- 4) 2 ) 4 ) 9 ) DS( 44 ) DG( 11- 17) DG( 18- 21) DG( 16- 21) DS( 40- 44) DG( 0- 7) 10982 10983 10984 1.6 MG/KG PFOS + CHOLESTEROL DESCRIPTION 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/BKOKEN 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 I'ROTOC'OL418 018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLIE 23 (PAGE 13): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL RAT # DESCRIPTION 11572 1.1573 7.1574 11575 11576 11577 11578 11579 11580 11581 11582 11583 11584 11585 DL( DL( 3 ) 3 ) DG( 1- 2 ) DL( 3 ) DL( 2 ) DG( 18- 2 1 ) DL( 1- 2) DG( 12- 19) DL( 5 ) SACRIFICED DUE TO NO SACRIFICED DUE TO NO NO ADVERSE FINDINGS CHROMODACRYORRHEA NO ADVERSE FINDINGS SACRIFICED DUE TO NO NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: SACRIFICED DUE TO NO SURVIVING SURVIVING SURVIVING SURVIVING LIMBS LIMBS LIMBS SURVIVING PUPS PUPS PUPS PUPS PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION PROTOCOL, 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 14): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP I 2 MG/KG PFOS + CHOLESTEROL RAT # DESCRIPTION 10985 NO ADVERSE FINDINGS 10986 2- 3) LOCALIZED ALOPECIA: LIMBS 18- 20) LOCALIZED ALOPECIA: LIMBS 1- 3) LOCALIZED ALOPECIA: LIMBS a 3) SACRIFICED DUE TO NO SURVIVING PUPS 10987 NO ADVERSE FINDINGS 10988 NO ADVERSE FINDINGS 10989 NO ADVERSE FINDINGS 10990 NO ADVERSE FINDINGS 10991 NO ADVERSE FINDINGS 10992 16 ) CHROMORHINORRHEA 10993 1- 4) CHROMODACRYORRHEA 20- 21) INCISORS: MISSING/BROKEN a 10994 3) SACRIFICED DUE TO NO SURVIVING PUPS 10995 NO ADVERSE FINDINGS 10996 2) SACRIFICED DUE TO NO SURVIVING PUPS 10997 18 ) CHROMORHINORRHEA 10998 NO ADVERSE FINDINGS 11586 3) SACRIFICED DUE TO NO SURVIVING PUPS 11587 3) SACRIFICED DUE TO NO SURVIVING PUPS 11588 2) SACRIFICED DUE TO NO SURVIVING PUPS 11589 10.- 16) INCISORS: MISSING/BROKEN 12- 15) CHROMODACRYORRHEA 5) SACRIFICED DUE TO NO SURVIVING PUPS 11590 15- 20) LOCALIZED ALOPECIA: LIMBS 1- 3 ) LOCALIZED ALOPECIA: LIMBS a 3) SACRIFICED DUE TO NO SURVIVING PUPS 11591 2) SACRIFICED DUE TO NO SURVIVING PUPS P c. b 11592 NO ADVERSE FINDINGS 00 11593 NO ADVERSE FINDINGS 11594 23- 34) LOCALIZED ALOPECIA: LIMBS c. 2- 20) LOCALIZED ALOPECIA: LIMBS 1- 5) LOCALIZED ALOPECIA: LIMBS a 5) SACRIFICED DUE TO NO SURVIVING PUPS PKO'TOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 15): CLINICAT, OBSERVATIONS - INDIVIDUAIJ DATA - Fo GENERATION FEMALE RATS RAT # 11595 11596 11597 11598 11599 DL( DG( DL( DL( DG( DG( DL( DL( DS( DL( DL( 1 ) 19- 20) 1- 3) 3 ) 1- 2 2 ) 13- 22) 1- 4 ) 1- 4) 37- 3 8 ) 3 ) 2 ) DESCRIPTION ____.___._.___.__.._____________________---------. SACRIFICED DUE TO NO SURVIVING LOCALIZED ALOPECIA: BACK LOCALIZED ALOPECIA: BACK a SACRIFICED DUE TO NO SURVIVING LOCALIZED ALOPECIA: UNDERSIDE LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: UNDERSIDE INCISORS: MISSING/BROKEN SACRIFICED DUE TO NO SURVIVING SACRIFICED DUE TO NO SURVIVING PUPS PUPS PUPS PUPS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION IJKO'T'OCOI, 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 1 6 ) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP 8 0.4 MG/KG PFOS ------ RAT # DESCRIPTION ....- .. .. ..- .._.. ........ ....... .. .._ . . . . _ . . _ _ . _ . _ _ . . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ~ ~ ~ ~ 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 DS( 14- 3 8 ) DS( 39- 44) DG( 0- 20) DL( 1- 5) DG( 15 ) DG( 15- 21) DL( 1 DS( 20- 27) DS( 23- 27) DS( 18- 42) DG( 0- 20) DL( 1- 5) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS FOREPAWS AND LEFT FORELIMB: SCABS (EACH 0.1 CM IN DIAMETER) 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 DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL,418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 17): CLINICAL OBSERVATIONS - 1NDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP 9 0.8 MG/KG PFOS .................................................................................................................................... RAT # DESCRIPTION 11009 NO ADVERSE FINDINGS 11010 NO ADVERSE FINDINGS 11011 NO ADVERSE FINDINGS 11012 NO ADVERSE FINDINGS 11013 NO ADVERSE FINDINGS 11014 DS( 32- 34) INCISORS: MISSING/BROKEN 11015 NO ADVERSE FINDINGS 11016 NO ADVERSE FINDINGS 11017 NO ADVERSE FINDINGS 11018 NO ADVERSE FINDINGS 11610 NO ADVERSE FINDINGS 11611. NO ADVERSE FINDINGS 11612 NO ADVERSE FINDINGS 11613 NO ADVERSE FINDINGS 11614 NO ADVERSE FINDINGS 11.615 NO ADVERSE FINDINGS 11616 NO ADVERSE FINDINGS 11617 NO ADVERSE FINDINGS 11610 NO ADVERSE FINDINGS 11619 DS( 10.- 20) CHROMODACRYORRHEA DS( 18- 42) INCISORS: MISALIGNED DS( 27- 42) CHROMODACRYORRHEA DG( 0- 20) CHROMODACRYORRHEA DG( 0 - 20) INCISORS: MISALIGNED DG( 13- 19) INCISORS: MISSING/BROXEN DL( 1- 2) INCISORS: MISALIGNED DL( 1.- 5) CHROMODACRYORRHEA a DL( 2- 3) LACRIMATION p. c. b DL( 2- 5) INCISORS: MISSING/BROKEN a 00 .................................................................................................................................... DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION + a . Observation corif irmed at necropsy . PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 18): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11019 14- 20) LOCALIZED ALOPECIA: LIMBS 1- 4 ) LOCALIZED ALOPECIA: LIMBS 11020 NO ADVERSE FINDINGS 11021 NO ADVERSE FINDINGS 11022 NO ADVERSE FINDINGS 11023 13- 18) LOCALIZED ALOPECIA: UNDERSIDE 11024 38- 42) LOCALIZED ALOPECIA: LIMBS 0- 21) LOCALIZED ALOPECIA: LIMBS 1- 4) LOCALIZED ALOPECIA: LIMBS 11025 NO ADVERSE FINDINGS 11026 NO ADVERSE FINDINGS 11027 NO ADVERSE FINDINGS 11028 34- 44) LOCALIZED ALOPECIA: LIMBS 0- 20) LOCALIZED ALOPECIA: LIMBS 1- 5 ) LOCALIZED ALOPECIA: LIMBS a 11620 NO ADVERSE FINDINGS 11621 NO ADVERSE FINDINGS 11622 NO ADVERSE FINDINGS 11623b NO ADVERSE FINDINGS 11624 NO ADVERSE FINDINGS 11625 NO ADVERSE FINDINGS 11626 5- 21) LOCALIZED ALOPECIA: UNDERSIDE 1- 4 ) LOCALIZED ALOPECIA: UNDERSIDE 11627 3) SACRIFICED DUE TO NO SURVIVING PUPS 11628 2) SWOLLEN SNOUT 2- 10) INCISORS: MISSING/BROKEN 11629 NO ADVERSE FINDINGS - . - - - - . _ _ _ _ - - _ _ _ . . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - . . - - - - - - - - - - - - - - . -P- - - - - CL 90 DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. Observation confirmed at necropsy. 0 b. Rat 11623 was missing and found on day 19 of study. CL m M PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 3 (PAGE 1 9 ) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 DS( 1 7 - 4 3 ) DG( 0 - 2 0 ) DL( 1- 5 ) DG( 11- 2 0 ) DL( 1- 4 ) DG( 11- 2 0 ) DG( 1 9 - 2 0 ) DG( 1 9 - 2 0 ) DL( 1- 4 ) DLt( 1- 4 ) DL( 1- 4 ) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LIMBS LIMBS LIMBS a BACK BACK UNDERSIDE LIMBS BACK LIMBS UNDERSIDE BACK DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. DL = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 20): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 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 11 652 11653 2) 5- 21) 37- 44) 0- 3) 2) 10- 19) 11- 44) 0- 21) 1- 5 ) 11.- 2 0 ) 1- 5 ) 0- 20) 12- 1 7 ) 1- 5) 5) 2) 5) 11- 13) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING 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 LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: NECK LOCALIZED ALOPECIA: LIMBS a LOCALIZED ALOPECIA: NECK a SACRIFICED DUE TO NO SURVIVING NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS SACRIFICED DUE TO NO SURVIVING NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN NO ADVERSE FINDINGS PUPS a PUPS a PUPS PUPS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 23 (PAGE 21): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ....--_______ RAT # _________.__. 11053 NO ADVERSE FINDINGS 11054 NO ADVERSE FINDINGS 11055 DL( 4 ) SACRIFICED DUE TO NO SURVIVING PUPS 11056 NO ADVERSE FINDINGS 11057 DL( 2 ) SACRIFICED DUE TO NO SURVIVING PUPS 11058 NO ADVERSE FINDINGS 11059 NO ADVERSE FINDINGS 11060 NO ADVERSE FINDINGS 11061 D L ( 3 ) SACRIFICED DUE TO NO SURVIVING PUPS 11062 NO ADVERSE FINDINGS 11063 NO ADVERSE FINDINGS 11064 DL( 3 ) SACRIFICED DUE TO NO SURVIVING PUPS 11065 DL( 2 ) SACRIFICED DUE TO NO SURVIVING PUPS 11066 NO ADVERSE FINDINGS 11654 NO ADVERSE FINDINGS 11655 DG( 17- 21) LOCALIZED ALOPECIA: LIMBS DG( 1 7 - 21) LOCALIZED ALOPECIA: NECK DL( 1- 2) LOCALIZED ALOPECIA: LIMBS DL( 1- 5) LOCALIZED ALOPECIA: NECK a 11656 NO ADVERSE FINDINGS 11657 D S ( 17- 43) LOCALIZED ALOPECIA: LIMBS DG( 0 - 20) LOCALIZED ALOPECIA: LIMBS DL( 1- 3) LOCALIZED ALOPECIA: LIMBS a DL( 3 ) SACRIFICED DUE TO NO SURVIVING PUPS 11658 DL( 2 ) SACRIFICED DUE TO NO SURVIVING PUPS 11659 DG( 1 2 - 21) LOCALIZED ALOPECIA: LIMBS DL( 1 ) LOCALIZED ALOPECIA: LIMBS a P - DL( 1 ) SACRIFICED DUE TO NO SURVIVING PUPS 00 b 11660 DL( 3 ) SACRIFICED DUE TO NO SURVIVING PUPS - 1 1 6 6 1 DL( 2 ) SACRIFICED DUE TO NO SURVIVING PUPS 11662 NO ADVERSE FINDINGS 11663 SACRIFICED DUE TO NO SURVIVING PUPS 11664 SACRIFICED DUE TO NO SURVIVING PUPS m 11665 SACRIFICED DUE TO NO SURVIVING PUPS 11 11 66 66 67 NSOACARDIVFEIRCSEED DFUINEDTIONGNSO SURVIVING PUPS W c-. 0 c.L DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a ___________..________________ 1 VEHICLE CONTROL 10901 DG 21 P ALL TISSUES APPEARED NORMAL. 10902 DG 21 P ALL TISSUES APPEARED NORMAL. 10903 DG 21 P ALL TISSUES APPEARED NORMAL. 10904 DG 21 P ALL TISSUES APPEARED NORMAL. 10905 DG 21 P ALL TISSUES APPEARED NORMAL. 10906 DL 5 P ALL TISSUES APPEARED NORMAL. 10907 DG 21 P ALL TISSUES APPEARED NORMAL. 10908 DG 21 P ALL TISSUES APPEARED NORMAL. 10909 DL 5 P ALL TISSUES APPEARED NORMAL. 10910 DG 21 P ALL TISSUES APPEARED NORMAL. 10911 DL 5 P ALL TISSUES APPEARED NORMAL. 10912 DG 21 NP ALL TISSUES APPEARED NORMAL. 10913 DL 5 P ALL TISSUES APPEARED NORMAL. 10914 DL 5 P ALL TISSUES APPEARED NORMAL. 11501 DL 5 P ALL TISSUES APPEARED NORMAL. 11502 DL 5 P ALL TISSUES APPEARED NORMAL. 11503 DL 5 P ALL TISSUES APPEARED NORMAL. 11504 DL 5 P ALL TISSUES APPEARED NORMAL. 11505 DL 5 P ALL TISSUES APPEARED NORMAL. 11506 DL 5 P ALL TISSUES APPEARED NORMAL. 11507 DL 5 P ALL TISSUES APPEARED NORMAL. 11508 DL 5 P ALL TISSUES APPEARED NORMAL. 11509 DL 5 P ALL TISSUES APPEARED NORMAL. 11510 DL 5 P ALL TISSUES APPEARED NORMAL. 11511 DL 5 P ALL TISSUES APPEARED NORMAL. 11512 DL 5 P ALL TISSUES APPEARED NORMAL. 11513 DL 5 P ALL TISSUES APPEARED NORMAL. 11514 DL 5 P 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 2 MEVALONIC ACID CONTROL 10915 10916 10917 10918 10919 10920 10921 10922 10923 10924 10925 DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DL 5 P DL 5 P DL 5 P DL 5 P DG 21 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10926 DG 12 P MORIBUND SACRIFICED ON DAY 12 OF GESTATION (DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT). ESOPHAGUS: PERFORATION (0.2 CM X 0.3 CM) . LEFT AXILLARY AREA: TISSUE DISCOLORED TAN. UTERINE CONTENTS: 14 IMPLANTATION SITES (14 LIVE EMBRYOS).b ALL OTHER TISSUES APPEARED NORMAL. 10927 10928 11515 11516 11517 11518 11519 DL 5 DG 21 DL 5 DL 5 DL 5 DL 5 DL 5 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 DL 5 P ALL TISSUES APPEARED NORMAL. 11522 DG 25 NP 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 3 ) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ...__._____..._.._._~~~~~~~~.~~~~~~..--~~~~..... DOSAGE GROUP DOSAGE MG/KG PFOS wr NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a ._.._.._._-____..._.____________________~~~..~.. 2 (CONT.) MEVALONIC ACID CONTROL 11523 DL 5 ALL TISSUES APPEARED NORMAL. 11524 DL 5 ALL TISSUES APPEARED NORMAL. 11525 DL 5 ALL TISSUES APPEARED NORMAL. 11526 DL 5 ALL TISSUES APPEARED NORMAL. 11527 DL 5 ALL TISSUES APPEARED NORMAL. 11528 DL 5 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 2 3 ) for external observations confirmed at necropsy. PROTOCOL,4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 4 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 3 1 . 6 t MEVALONIC ACID 10929 10930 10931 10932 10933 10934 10935 DL 5 P DG 2 1 P DG 2 1 P DG 2 1 P DG 2 1 P DG 2 1 P DG 2 1 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10936 DS 2 FOUND DEAD ON DAY 2 OF STUDY (DEATH OCCURRED 5 8 MINUTES AFTER THE 2NE DOSAGE ADMINISTRATION). THORACIC CAVITY: CONTAINED DARK RED CLOTTED MATERIAL. ALL OTHER TISSUES APPEARED NORMAL. 10937 10938 10939 10940 DG 2 1 P DL 5 P DG 2 5 NP DG 2 1 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10941. DL 4 P SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 10942 11529 11530 11531 DL 5 P DL 5 P DI, 5 P DG 2 5 NP ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11532 11533 DL 2 P DL 3 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. b 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 2 3 ) for external observations confirmed at necropsy PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 5): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS -.-_____________----____________________--~~~.~.~~~..~ DOSAGE GROUP RAT DAY OF PREGNANCY _ _ DOSAGE MG/KG PFOS NUMBER NECROPSY STATUS OBSERVATIONS a - - . - - _ _ _ - - _ _ _ _ . _ _ _ _ _ _ _ - - _ _ _ _ _ _ _ _ _ -_ . _ __ __ _._ __ __ __ __ __ __ __ __ _-^ - _ . ._ . -_ ~_ _ _ _ _ _ _ _ _ _ _ _ _ _ 3 (CONT. ) 1.6 + MEVALONIC ACID 11536 DL 5 P ALL TISSUES APPEARED NORMAL. 11537 DL 5 P ALL TISSUES APPEARED NORMAL. 11538 DL 5 P 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 P DL 5 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. W PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 6): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER 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 P DG 21 P DG 21 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 P DG 21 P DG 21 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 P DG 21 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10955 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 10956 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11544 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11545 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11546 DL 1 w c.' 0 4 DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P = PREGNANT NP = NOT PREGNANT d . Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. PKorocoL,418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLtE 24 (PAGE ' I ) : NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP RAT DOSAGE MG/KG PFOS NUMBER .____._........_..._..~.~....~...~. 4 (CONT.) 2 + MEVALONIC ACID 11547 11548 DL 5 P DG 25 NP OBSERVATIONS a ____________________________ 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. 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. w 0 00 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 8): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 5 CHOLESTEROL CONTROL RAT NUMBER 10957 10958 10959 10960 10961 10962 10963 10964 10965 10966 10967 10968 10969 10970 11558 11559 11560 11561 11562 DAY OF PREGNANCY NECROPSY STATUS DL 5 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DG 25 NP DL 5 P DS 16 11563 11564 11565 11566 11567 11568 11569 11570 11571 DL 5 P DL 5 P DL 5 P DG 25 NP DL 5 P DL 5 P DL 5 P DL 5 P DL 5 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. MORIBUND SACRIFICED ON DAY 16 OF STUDY. SMALL INTESTINES: TWISTED AND CONSTRICTED AT JEJUNUM JEJUNUM: CONTAINED DARK RED FLUID. ALL OTHER TISSUES APPEARED NORMAL. - ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 9): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 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 P DG 21 P DG 21 P DL 5 P DG 21 P DL 5 P DG 21 P DG 21 P DG 21 P DL 5 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10984 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11572 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11573 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11574 11575 11576 DL 5 P DL 5 P DL 5 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 10): NECROPSY OBSERVATIONS - 1NDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGE MG/KG PFOS NUMBER NECROPSY STATUS OBSERVATIONS a ..._-.--___.____._-_____________________~~~------- 6 (CONT.) 1.6 + CHOLESTEROL 11578 11579 11580 DL 5 P DL 5 P DL 5 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 P DL 5 P DL 5 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. PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 11): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a DG 21 P ALL TISSUES APPEARED NORMAL 10986 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10987 10988 10989 10990 10991 10992 10993 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 DG 21 ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 10994 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 10995 DL 5 P ALL TISSUES APPEARED NORMAL. 10996 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. STOMACH: CONTAINED PUP TISSUE. ALL OTHER TISSUES APPEARED NORMAL. 10997 10998 DL 5 P DG 25 NP ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11586 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11587 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11588 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11589 DL 5 P SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. w . _ _ _ . _ . _ _ _ _ _ _ _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - - - - - - . - - - . - - L -_r- - - - - C-L DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION N P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 12): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 7 (CONT.) 2 + CHOLESTEROL RAT NUMBER 11590 ..-.._____ DAY OF NECROPSY ____._____ 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 P DL 5 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. US: = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION P b P = PREGNANT NP = NOT PREGNANT C-L 00 a . Refer to the individual clinical observations table (Table 23) for external observations confirmed at necropsy. C-L m.o~oCoL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) T m L E 24 (PAGE 13): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP RAT DAY OF DOSAGE MG/KG PFOS NUMBER NECROPSY ____._____...._.....~~.~.~...~~~.~. 8 0.4 10999 DL 5 P ALL TISSUES APPEARED NORMAL. 11000 DL 5 P ALL TISSUES APPEARED NORMAL. 11001 DL 5 P ALL TISSUES APPEARED NORMAL. 11002 DL 5 P ALL TISSUES APPEARED NORMAL. 11003 DL 5 P ALL TISSUES APPEARED NORMAL. 11004 DL 5 P ALL TISSUES APPEARED NORMAL. 11005 DL 5 P ALL TISSUES APPEARED NORMAL. 11006 DL 5 P ALL TISSUES APPEARED NORMAL. 11007 DL 5 P ALL TISSUES APPEARED NORMAL. 11008 DL 5 P ALL TISSUES APPEARED NORMAL. 11600 DL 5 P ALL TISSUES APPEARED NORMAL. 11601 DL 5 P ALL TISSUES APPEARED NORMAL. 11602 DL 5 P ALL TISSUES APPEARED NORMAL. 11603 DL 5 P ALL TISSUES APPEARED NORMAL. 11604 DL 5 P ALL TISSUES APPEARED NORMAL. 11605 DL 5 P ALL TISSUES APPEARED NORMAL. 11606 DL 5 P ALL TISSUES APPEARED NORMAL. 11607 DL 5 P ALL TISSUES APPEARED NORMAL. 11608 DL 5 P ALL TISSUES APPEARED NORMAL. 11609 DL 5 P 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. P L3 cp 0 L3 00 0 m c -P PRO'I'OCOL 418-018: 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 - F O GENERATION FEMALE RATS . - . . . - _ _ _ _ _ __._ . _ _ _ . . __ ___ _ _ - _ . _ - ._ - - _ . _ . _ _ _ _ _ _ _ _ _ _ _ . . - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGE MG/KG PFOS NUMBER NECROPSY STATUS OBSERVATIONS a _._....._._______..._______________ 9 0.8 11009 11010 11011 11012 11013 11014 11015 11016 11017 11018 11610 11611 11612 11613 11614 11615 11616 11617 11618 11619 DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DS 81 NP DL 5 P DL 5 P DG 25 NP DL 5 P DG 25 NP DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 15): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE MG/KG PFOS 10 1 RAT NUMBER 11019 11020 11021 11022 11023 11024 11025 11026 11027 11028 11620 11621 11622 11623 11624 11625 11626 DAY OF PREGNANCY NECROPSY STATUS __________-_-______ DL 5 P DL 5 P DL 5 P DG 25 NP DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P DL 5 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11627 DL 3 F SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11628 11629 DL 5 P DL 5 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. PrivrocoL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 16): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS RAT DAY OF PREGNANCY NUMBER NECROPSY STATUS ...__..-_.______.--_____________________- LL 1.2 11029 DL 5 P ALL TISSUES APPEARED NORMAL. 11030 DS 81 NP ALL TISSUES APPEARED NORMAL. 11031 DL 5 P ALL TISSUES APPEARED NORMAL. 11032 DL 5 P ALL TISSUES APPEARED NORMAL. 11033 DL 5 P ALL TISSUES APPEARED NORMAL. 11034 DL 5 P ALL TISSUES APPEARED NORMAL. 11035 DL 5 P ALL TISSUES APPEARED NORMAL. 11036 DL 5 P ALL TISSUES APPEARED NORMAL. 11037 DL 5 P ALL TISSUES APPEARED NORMAL. 11038 DL 5 P ALL TISSUES APPEARED NORMAL. 11630 DL 5 P ALL TISSUES APPEARED NORMAL. 11631 DG 25 NP ALL TISSUES APPEARED NORMAL. 11632 DL 5 P ALL TISSUES APPEARED NORMAL. 11633 DL 5 P ALL TISSUES APPEARED NORMAL. 11634 DL 5 P ALL TISSUES APPEARED NORMAL. 11635 DL 5 P ALL TISSUES APPEARED NORMAL. 11636 DL 5 P ALL TISSUES APPEARED NORMAL. 11637 DL 5 P ALL TISSUES APPEARED NORMAL. 11638 DL 5 P ALL TISSUES APPEARED NORMAL. 11639 DL 5 P 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. bc-' PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 17): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DOSAGE GROUP RAT DAY OF DOSAGE MG/KG PFOS NUMBER NECROPSY . - - _ _ _ _ _ _ _ _ _ _ _ _ _ - - -.- . - -.- _ - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ 12 1.6 11039 11040 11041 DS 81 NP DG 21 P DL 5 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11042 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11043 11044 11045 11046 11047 DG 25 NP DG 21 P DG 21 P DG 21 P DG 21 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11048 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11049 11050 11051 11052 11640 11641 11642 11643 11644 DL 5 DG 21 DG 21 DG 21 DL 5 DL 5 DL 5 DL 5 DL 5 ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11645 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11646 11647 11648 DL 5 P DL 5 P DL 5 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 ('Table23) for external observations confirmed at necropsy. PROTOCOL 418-0163 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 18): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - F O GENERATION FEMALE RATS ..-__...__ __ _ _....- ..- _ _ _ _ _ . - - _ _ - - - - ~ ~ . - . . - ~ ~ ~ . . . - ~ - ~ ~ ~ ~ - - ~ - ~ . ~ ~ ~ - - - - - - - DOSAGE GROUP DOSAGE MG/KG PFOS RAT NUMBER DAY OF PREGNANCY NECROPSY STATUS OBSERVATIONS a 12 (CONT.) 1.6 11650 11651 11652 11653 DL 5 P DL 5 P DG 25 NP DL 5 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. US = 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR`SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 19): NECROPSY OBSERVATlONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS .. DG 21 P 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 P DL 5 P DG 21 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 P DG 21 P ALL TISSUES APPEARED NORMAL. ALL TISSUES APPEARED NORMAL. 11064 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11065 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ALL TISSUES APPEARED NORMAL. 11066 11654 11655 11656 DG 21 P DL 5 P DL 5 P DL 5 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 O F 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) f o r external observations confirmed at necropsy PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 24 (PAGE 20): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGE MG/KG PFOS NUMBER NECROPSY STATUS OBSERVATIONS a . . _ _ _ _ _ _ _ _ _ _ . _ . __ . _ __ _._ . - _._ - -_ - _ - -_ - - . _ - -_ - . _ __ __ __ __ _- _._ - - _ -.-_ . ._ -_ - -_ -_ - -_ -_ . ._ _ _ _ _ _ _ _ _ _ _ _ 13 (CONT.) 2 11659 DL 1 P SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11660 DL 3 P SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11661 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11662 DL 5 P ALL TISSUES APPEARED NORMAL. 11663 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11664 DL 2 P SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS. ALL TISSUES APPEARED NORMAL. 11665 DL 5 P SACRIFICED ON DAY 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 10901 P 405. C 14.15 10902 P 389. C 12.81 10903 P 436. C 14.28 10904 P 420. c 15.04 10905 P 401. C 14.84 10906 P 344. D 15.66 10907 P 430. C 14.33 10908 P 417. C 14.86 10909 P 294. D 11.82 10910 P 442. C 14.30 10911 P 300. D 15.67 10912 NP 338. C 12.49 10913 P 299. D 11.40 10914 P 280. D 11.76 11501 P 261. D 12.19 11502 P 328. D 13.38 11503 P 310. D 12.63 11504 P 347. D 14.39 11505 P 319. D 11.40 11506 P 347. D 17.55 11507 P 336. D 14.81 11508 P 328. D 12.54 11509 P 342. D 13.93 11510 P 359. D 14.09 11511 P 299. D 12.10 11512 P 357. D 11.77 11513 P 279. D 11.45 11514 P 327. D 12.20 - - .- - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 DOSAGE GROUP 2 _ . _ _ . . _ _._ . _ _ ..._ ._ . _ _ _ . _ _ 10915 P 402. C 12.81 10916 P 452. C 14.86 10917 P 383. C 12.74 10918 P 443. C 16.58 10919 P 391. C 14.34 10920 P 426. C 19.20 10921 P 317. D 13.24 10922 P 321. D 13.04 10923 P 296. D 12.43 10924 P 301. D 12.51 10925 P 10926a P 4_ 2_ 8_ . C 16.03 10.04 10927 P 341. D 13.36 10928 P 408. C 16.03 11515 P 282. D 11.82 11516 P 341. D 13.94 11517 P 328. D 14.98 11518 P 337. D 13.14 11519 P 306. D 11.82 11520 P 303. D 13.49 11521 P 341. D 13.05 11522 NP 11523 P 313. D 14.51 4.64 P - 11524 P 305. D 12.10 3.97 03 11525 P 333. D 13.46 4.04 b 11526 P 314. D 12.89 4.10 c-' 11527 P 348. D 16.72 4.80 PP 11528 P 324. D 12.29 3.79 Td 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 10926 was moribund sacrificed on day 12 of gestation; values excluded from group averages and statistical analyses. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 ___. 1.6 MG/KG PFOS + MEVALONIC ACID 10929 P 284. D 13.55 10930 P 386. C 15.78 10931 P 382. C 14.11 10932 P - _ _ C a 10933 P 392. C 16.00 10934 P 391. C 19.10 10935 P 363. C 12.81 10936b - _ _ _ L O . 59 10937 P 383. C 17.21 10938 P 316. D 15.82 10939 NP 10940 P 459. C 18.76 1 0 9 4 1 ~P _ _ _ 12.38 4.09 __.. 10942 P 311. D 16.18 5.20 11529 P 280. D 12.71 4.54 11530 P 355. D 15.07 4.24 11531 NP 1 1 5 3 2 ~P _ _ _ 17.59 1 1 5 3 3 ~P - _ _ 12.63 11534 P 336. D 15.30 1 1 5 3 5 ~P _ _ _ 15.27 4.55 ____ 11536 P 320. D 15.63 4.88 11537 P 308. D 15.01 4.87 11538 P 1 1 5 3 9 ~P 304. D .__ 13.25 10.86 4.36 ____ 11540 P 11541C P 330. D ___ 16.51 13.76 5.00 _.__ 11542 P 298. D 13.73 4.61 11543 P 348. D 15.20 4.37 _ _ _ . _ __ _ _ __ _ _ _ __ __ . _ .. . _ _ ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. w C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION c-. a. Value was not recorded. t3 P 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 WEIGHI' LIVER ABS . REL. ______._.___ 10943a P 14.51 __._ 10944 P D 11.92 4.23 10945 P C 16.51 3.98 10946 P C 17.22 4.09 10947 P 10948a P C 14.26 18.07 3_ _._7_0 10949 P C 12.70 4.54 10950 P C 16.49 3.92 10951 P C 16.95 4.06 10952a P 14.33 _--_ 10953 P C 16.16 4.05 10954 P 10955a P C 16.04 16.30 3_ _._9_4 10956a P 16.88 11544a P 13.74 11545a P 1154Ga P 18.81 14.56 ____ 11547 P D 14.89 4.98 11548 NP 11549a P 17.01 11550a P 13.74 b 11551a P 15.29 -P 11552 P D 14.55 w 00 11553d P 16.80 11554a P 13.22 c-. 11555a P 13.15 11556a P 13.73 11557a P 16.09 -.--___..___ 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) 'TABLE 25 (PAGE 5): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - FO GENERATION FEMALE RATS NUMBER WEIGHT ABS. WT. REL. % TBW . _ _ _ _ _ _ _ _ _ _ _ _ _ _ 10957 P 316. D 12.06 10958 P 448. C 14.99 10959 P 448. C 14.08 10960 P 442. C 15.88 10961 P 409. C 12.03 10962 P 444. C 12.78 10963 P 475. C 14.61 10964 P 476. C 14.81 10965 P 386. C 13.47 10966 P 379. D 15.15 10967 P 10968 P 326. D 319. D 12.17 12.62 10969 P 10970 P 320. D 323. D 11.57 15.17 11558 P 11559 P 308. D 340. D 13.95 15.26 11560 NP 11561 P 11562a - 3_ 5_ _9. D 14.21 b 11563 P 325. D 13.87 11564 P 306. D 11.71 11565 P 321. D 13.18 11566 NP 11567 P 347. D 14.07 11568 P 312. D 14.46 11569 P 355. D 13.75 11570 P 242. D 10.88 11571 P 379. D 14.48 3.96 4.27 3.83 4.10 4.05 4.63 3.87 4.50 3 .a2 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. Rat 11562 was moribund sacrificed on day 16 of study; values excluded from group averages and statistical analyses. b. Value was not recorded. F'ROTOCOI,418-018: 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 F!ATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT TERMINAL BODY NUMBER WEIGHT 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. C ___ 435. C 339. C 376. C 269. D 415. C 316. D 402. C 373. C 429. C 299. D __- ___ ___ 284. D 335. D 307. D ___ 291. D 325. D 319. D _-- 298. D 317, D 309. D 282. 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 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABL,E 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 WEIGH? LIVER ABS. WT. REL. % TBW . - - - _ _ _ . _ __ ..._ _ __ _ ._ _ . . _ 10985 P 10986a P 4_ 0_ 2_ . C 15.39 15.35 10987 P 394. C 15.90 10988 P 397. C 14.96 10989 P 323. C 13.91 10990 P 403. C 15.27 10991 P 379. C 15.47 10992 P 372. C 16.30 10993 P 430. C 15.14 10994a P _ _ . 15.64 10995 P 282. D 13.99 10996a P _ _ _ 16.60 10997 P 261. D 11.28 10998 NP 11586a P _ - 11587a P _ _ _ 11588a P _ _ _ 12.74 14.88 12.53 11589 P 290. D 12.24 11590d P 11591a P _.. ___ 14.76 17.35 11592 P 297. D 13.14 11593 P 262. D 12.43 90 11594 P 299. D 13.24 -P 11595a P _ _ _ 16.06 __. 11596a P _ - . 13.75 0 11597 P 275. D 12.10 11598a P _ - - 13.25 11599a P _ - - 16.71 . _ _ _ _ _ __ __ - _ - ._ . . ALL WEIGHTS WERE RECORDED IN GRRMS (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. E'ROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 10999 P 306. D 12.95 4.23 11000 P 333. D 11.72 3.52 11001 P 304. D 13.06 4.30 11002 P 325. D 14.78 4.55 11003 P 333. D 13.93 4.18 11004 P 322. D 14.31 4.44 11005 P 325. D 14.76 4.54 11006 P 334. D 14.91 4.46 11007 P 340. D 16.47 4.84 11008 P 325. D 14.96 4.60 11600 P 344. D 15.30 4.45 11601 P 383. D 16.13 4.21 11602 P 281. D 10.83 3.85 11603 P 333. D 13.32 4.00 11604 P 297. D 11.89 4.00 11605 P 337. D 12.79 3.80 11606 P 312. D 12.46 3.99 11607 P 312. D 13.39 4.29 11608 P 345. D 14.17 4.11 11609 P 343. D 14.65 .- ...- 4.27 ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) REL. % TBW = (ORGANWEIGHT/TERMINAL BODY WEIGHT) x 1 0 0 C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION b PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 0.8 MG/KG PFOS 11009 P 208. D 12.16 5.85 11010 P 346. D 15.03 4.34 11011 P 335. D 14.14 4.22 11012 P 283. D 13.00 4.59 11013 P 290. D 13.78 4.75 11014 NP 11015 P 298. D 13.64 4.58 11016 P 279. D 11.15 4.00 11017 NP 11018 P 324. D 14.81 4.57 11610 NP 11611 P 356. D 14.16 3.98 11612 P 296. D 13.88 4.69 11613 P 364. D 14.51 3.99 11614 P 326. D 14.95 4.58 11615 P 355. D 14.09 3.97 11616 P 278. D 13.27 4.77 11617 P 326. D 13.45 4.12 11618 P 332. D 13.79 4.15 11619 P 245. D 12.04 4.91 .____________.__________________________--.-------------.---- ALL WEIGHTS WERE RECORDED IN GRAMS (G). ABS. WT. = ORGAN WEIGHT. REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100. Y P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) P C = CAESAREAN SECTIONED ON DAY 21 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION w 0 c. w 0 PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 5 (PAGE 10): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS LIVER RAT TERMINAL BODY NUMBER WEIGHT ABS . WT . .- - - - - - REL . % TBW _ _ _ _ DOSAGE GROUP 10 _ _ _ _._ __ _.._ _ _.-_ - - -_ - - 11019 P 306. D 14.75 11020 P 336. D 15.26 11021 P 282. D 15.71 1 1 0 2 2 NP 11023 P 281. D 14.01 4.98 11024 P 337. D 15.93 4.73 11025 P 314. D 13.15 4.19 11026 P 308. D 13.34 4.33 11027 P 277. D 11.86 4.28 11028 P 300. D 15.70 5.23 11620 P 275. D 12.79 4.65 11621 P 317. D 12.48 3.94 11622 P 322. D 12.46 3.87 11623 P 305. D 12.68 4.16 11624 P 323. D 14.63 4.53 11625 P 323. D 14 .49 4.49 11626 P 11627a P 310. _-- D 11.80 13.73 3.81 11628 P 324. D 12.33 11629 P 331. D 13.14 .- - - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES ABS. WT. = EXCLUDED FROM ORGAN WEIGHT. AVERAGES) REL. % TBW = (ORGAN WEIGHT/TERMINAL BODY WEIGHT) X 100 C = CAESAREAN SECTIONED ON DAY 2 1 OF PRESUMED GESTATION D = DELIVERED A LITTER AND SACRIFICED ON DAY 5 OF LACTATION b a. Dam was sacrificed due to no surviving pups prior to day 5 of lactation; values excluded from group averages and statistical analyses. r-L PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 25 (PAGE 11): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS m r TERMINAL BODY NUMBER WEIGHT LIVER 11029 P 304. D 11030 NP 11031 P 302. D 13.38 4.43 11032 P 321. D 14.72 4.58 11033 P 273. D 13.08 4.79 1.1034 P 296. D 14.24 4.81 11035 P 274. D 13.19 4.81 11036 P 338. D 17.69 5.23 11037 P 280. D 14.23 5.08 11038 P 306. D 15.44 5.04 11630 P 286. D 13.22 4.62 11631 NP 11632 P 287. D 12.77 4.45 11633 P 347. D 16.47 4.75 11634 P 329. D 13.80 4.19 11635 P 333. D 16.42 4.93 11636 P 314. D 14.47 4.61 11637 P 310. D 14.62 4.72 11638 P 276. D 12.29 4.45 11639 P 325. D 14.49 4.46 ___.____..______________________________------------------.------. AL,L 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 E'RO'I'OCOL418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 25 (PAGE 12): TERMINAL BODY WEIGHTS AND ORGAN WEIGHTS AND RATIOS ( % ) OF ORGAN WEIGHT TO TERMINAL BODY WEIGHT INDIVIDUAL DATA - Fo GENERATION FEMALE RATS LIVER RAT TERMINAL BODY NUMBER WEIGHT ABS. REL. WT. % TBW ___..._____.._______.-----------------------..------.---.----------------- DOSAGE GROUP 12 _ _ _ . . . _ _ _ .._ _ _ ._ . _ __ . . _ 11039 NP 11040 P 374. C 17.98 11041 P 11042a P 2_ _9_2. D 12.95 16.60 11043 NP 11044 P 397. C 14.06 11045 P 391. C 13.30 11046 P 431. C 15.09 11047 P 384. C 16.38 11048a P - _ _ 15.53 11049 P 304. D 14.72 11050 P 400. C 14.87 11051 P 377. C 11.94 11052 P 444. C 15.61 11640 P 293. D 13.66 11641 P 316. D 14.02 11642 P 290. D 13.57 11643 P 281. D 12.41 11644 P 288. D 13.88 11645a P _ _ _ 16.08 11646 P 316. D 13.89 11647 P 300. D 12.62 11648 P 279. D 12.08 11649 P 305. D 13.96 11650 P 289. D 11.67 11651 P 352. D 14.44 11652 NP 11653 P 276. D 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. PROTOCOL, 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 ABS . WT . . _ ..- - - - - . _ _.-.- ..- . 11053 P 11054 P C 14.13 C 14.45 11055a P 11056 P 11057a P 11058 P 13.29 C 13.63 15.41 C 16.58 3.94 11059 P 11060 P 11061a P 11062 P D 14.21 C 16.11 14,26 C 16.15 4.99 3.79 4.14 11063 P 11064a P 11065a P C 14.55 14.37 12.88 3.48 .__. ____ 11066 P C 16.63 3.79 11654 P 11655 P D 11.74 D 14.24 4 .32 4.86 11656 P 11657a P D 12.88 14.86 4.62 ..__ 11658a P 11659a P 12.91 14.87 11660a P 14.02 11661a P 13.80 11662 P D 12.13 11663a P 15.31 13.664.a P 12.32 11665 P D 13.16 11666 NP 11667a P 17.32 _._____-___ 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. PHOTOCOI, 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 1): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP 1 . . . . _ _ _ _ . . _ . . _ _ . _ . _ _ .... VEHICLE CONTROL . _ . . _ ._ _ _ __ _ _ . 10901 206. 214. 231. 242. 248. 252. 10902 227. 224. 232. 240. 242. 236. 10903 205. 237. 248. 257. 264. 269. 10904 219. 230. 230. 243. 260. 267. 10905 198. 211. 217. 228. 251. 258. 10906 214. 238. 250. 262. 266. 277. 10907 211. 233. 244. 247. 262. 277. 10908 217. 232. 234. 247. 259. 261. 10909 197. 219. 232. 243. 251. 260. 10910 212. 241. 252. 263. 266. 278. 10911 201. 227. 234. 244. 244. 252. 10912 217. 242, 250. 257. 269. 283. 10913 204. 219. 224. 227. 242. 251. 10914 213. 224. 231. 227. 231. 240. 11501 203. 203. 207. 207. 213. 216. 11502 229. 244. 263. 280. 284. 294. 11503 219. 221. 237. 234. 244. 253. 11504 228. 236. 243. 253. 262. 268. 11505 208. 222. 224. 240. 243. 251. 11506 234. 249. 265. 278. 278. 282. 11507 222. 234. 236. 244. 250. 262. 11508 221. 242. 260. 261. 260. 270. 11509 208. 219. 226. 248. 252. 259. 11510 232. 249. 260. 267. 278. 279. 11511 216. 218. 221. 233. 234. 237. 11512 234. 246. 256. 281. 288. 298. 11513 210. 222. 228. 228. 231. 246. P + 11514 225. 239. 244. 260. 276. 277. __.._._.._____... _ -- - . _ _ 'p 0 ALL WEIGHTS WERE RECORDED IN GRAMS (G). c-. DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. w PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 2): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10915 205. 218. 218. 234. 249. 10916 224. 250. 266. 270. 287. 10917 209. 228. 239. 245. 248. 10918 214. 234. 241. 247. 262. 10919 204. 223. 229. 231. 243. 10920 220. 240. 253. 257. 266. 10921 205. 223. 237. 255. 257. 10922 230. 224. 244. 255. 261. 10923 191. 209. 219. 232. 248. 10924 215. 241. 247. 255. 263. 10925 209. 229. 240. 249. 261. 10926 220. 233. 241. 260. 271. 10927 211. 229. 242. 247. 266. 10928 199. 223. 218. 233. 237. 1.1515 206. 205. 214. 220. 222. 11516 240. 254. 273. 280. 274. 11517 215. 229. 239. 246. 247. 11518 229. 238. 258. 271. 281. 11519 206. 214. 226. 247. 263. 11520 227. 227. 237. 237. 238. 11521 224. 240. 254. 264. 275. 11522 231. 234. 252. 256. 267. 11523 210. 223. 235. 249. 249. 11524 216. 224. 235. 236. 239. 11525 226. 238. 253. 261. 262. 11526 224. 232. 237. 241. 256. 11527 214. 230. 236. 244. 248. 11528 230. 236. 242. . . _ . _ . _ - _ _ - __ _ _ _ _ _ ._. ._. _ - 252. 267. _- ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitationI . b. First value recorded after cohabitation. 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. 10929 202. 214. 10930 217. 236. 10931 202. 222. 10932 207. 222. 10933 193. 208. 10934 217. 243. 10935 212. 221. 10936 213. FOUND 10937 204. 227. 10938 212. 246. 10939 202. 220. 10940 214. 234. 10941 204. 219. 10942 211. 235. 11529 202. 217. 11530 247. 259. 11531 231. 237. 11532 222. 238. 11533 209. 223. 11534 232. 255. 11535 217. 230. 11536 221. 238. 11537 214. 228. 11538 236. 245. 11539 217. 210. 11540 227. 247. 11541 214. 226. 11542 229. 238. 11543 225. 248. _ _ . _ _ _ . . __ . ._ . _ __-_ . _ 216. 247. 226. 234. 218. 255. 214. DEAD ON 237. 249. 227. 244. 223. 244. 229. 275. 254. 246. 241. 275. 232. 258. 234. 262. 210. 258. 229. 238. 259. 220. 228. 236. 259. 266. 270. 233. 235. 241. 245. 254. 257. 230. 238. 235. 259. 261. 261. 226. 232. 235. DAY 2 OF STUDY 244. 249. 259. 268. 273. 285. 231. 227. 236. 253. 263. 264. 231. 232. 240. 258. 259. 268. 230. 236. 243. 290. 298. 297. 256. 254. 251. 260. 263. 264. 244. 246. 251. 276. 283. 286. 248. 259. 255. 261. 262. 267. 244. 250. 257. 268. 258. 266. 212. 211. 216. 269. 280. 288. 244. 248. 249. 248. 250. 258. 278. 284. 291. _ _ ._ __ _. _ ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . DAY = DAY OF STUDY a . Last value recorded before cohabitation. b. First value recorded after cohabitation. 235. 276. 238. 243. 234. 267. 233. PROTOCOL 418-018: ONE GENERATZON REPRODUCTION STUDY OF PFOS - MEVALONIC AClD/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TAE3LE 26 (PAGE 4): BODY WEIGHTS - PREMATING - INDIVlDUAL DATA - FO GENERATION FEMALE RATS 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 195. 222. 214. 207. 204. 217. 210. 212. 200. 219. 205. 212. 200. 214. 204. 243. 218. 218. 207. 231. 212. 231. 214. 232. 221. 234. 213. 229. 214. 242. 238. 233. 221. 240. 226. 236. 228 I 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. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 249. 247. 258. 256. 274. 278. 266. 262. 238. 243. 274. 273. 255. 259. 263. 265. 261. 267. 262. 264. 241. 251. 266. 266. 246. 252. 275. 268. 256. 259. 304. 320. 257. 264. 273. 277. 245. 246. 271. 280. 241. 244. 275. 284. 275. 272. 263. 267. 254. 261. 284. 293. 250. 252. 291. 290. _.__ 287. L'ROTOCOI,4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE mrs 10957 10958 10959 10960 10961 10562 10963 10964 10965 10566 10967 10968 10969 10970 11558 11559 11560 11561 11562 11563 11564 11565 11566 11567 11568 11569 11570 11571 198. 226. 212. 208. 199. 220. 202. 208. 208. 229. 205. 212. 208. 207. 206. 230. 226. 220. 215. 231. 212. 218. 210. 230. 214, 230. 207. 224, 220. 235. 234. 222 I 212. 237. 233. 238. 218. 247. 226. 222. 232. 247. 220. 254. 238. 245. 227. 246. 232. 230. 224. 246. 221. 249. 220. 244. _____ 233. 257. 236. 219. 218. 251. 243. 256. 224. 253. 236. 235. 240. 260. 238. 252. 258. 257. 225. 264. 247. 230. 232. 256. 225. 266. 232. 255. . _ _ -_ . ._ _ . __ _._ _ _ 243. 250. 262. 273. 285. 294. 250. 263. 263. 237. 258. 257. 226. 236. 254. 266. 290. 291. 256. 263. 283. 268. 270. 288. 241. 252. 255. 270. 284. 296. 249. 263. 280. 238. 249. 263. 245. 260. 267. 269. 296. 290. 244. 247. 251. 265. 272. 284. 267. 278. 290. 277. 292. 285. MORIBUND SACRIFICED ON DAY 1 6 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 6): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS _.....__ . _ ..__ _ _ . _ _ _ _ 10971 195. 211. 218. 10972 219. 244. 256. 10973 208. 223. 226. 10974 214. 228. 234. 10975 206. 223. 238. 10976 215. 241. 252. 10977 206. 232. 236. 10978 216. 233. 247. 10979 200. 221. 238. 10980 211. 228. 228. 10981 209. 228. 234. 10982 212. 230. 228. 10983 207. 224. 237. 10984 218. 236. 248. 11572 196. 195. 202. 11573 234. 244. 249. 11574 222. 234. 244. 11575 224. 236. 250. 11576 215. 237. 246. 11577 233. 246. 254. 11578 222. 232. 242. 11579 233. 241. 263. 11580 214. 229. 243. 11581 232. 244. 250. 11582 215. 236. 242. 11583 222. 238. 244. 11584 215. 230. 246. 11585 226. 237. 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. 231. 274. 270. 242. 238. 264. 261. 255. 263. 266. 263. 243. 244. 260. 266. 264. 262. 251. 255. 248. 245. 248. 246. 243. 239. 268. 262. 210. 208. 273. 276. 253. 255. 272. 276. 265. 263. 276. 274. 246. 250. 288. 291. 258. 258. 272. 272. 258. 260. 257. 259. 269. 271. 253. 254. . _ _ . _ . _ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ALL, WElGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. PKO?'OCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 26 (PAGE 7): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .. . - - - - - - - - - - .... . _ _ _ _ ..-_ . .. .. _ _ . 10985 198. 219. 232. 240. 246. 250. 10986 221. 249. 264. 272. 270. 275. 10987 205. 230. 249. 254. 250. 260. 10988 209. 240. 253. 261. 263. 261. 10989 200. 220. 226, 230. 231. 245. 10990 222. 235. 251. 261. 278. 280. 10991 210. 222. 234. 246. 243. 246. 10992 211. 232. 236. 239. 244. 253. 10993 210. 224. 242. 257. 264. 261. 10994 212. 242. 255. 266. 260. 265. 10995 207. 222. 233. 247. 246. 246. 10996 214. 226. 239. 245. 242. 251. 10997 197. 218. 225. 227. 232. 229. 10998 206. 230. 249. 258. 262. 268. 11586 206. 219. 221. 240. 246. 248. 11587 241. 259. 270. 275. 279. 287. 11588 217. 242. 253. 256. 260. 260. 11589 222. 239. 251. 256. 259. 263. 11590 215. 222. 234. 243. 242. 241. 11591 240. 248. 246. 265. 276. 281. 11592 214. 236. 250. 247. 256. 255. 11593 224. 224. 222. 227. 224. 225. 11594 214. 235. 242. 254. 265. 266. 11595 228. 241. 253. 252. 252. 256. 11596 222. 231. 230. 238. 234. 231. 11597 226. 246. 247, 252. 254. 248. 11598 215. 231. 237. 247. 243. 249. 11599 225. 242. 246. 251. 248. 254. . _ _ . - __ __ _ _ _ _ _ _ _ _ _ __ . . _ _ _ .- - _ - - -.- _ _ _ . ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 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. ____ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 8): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10999 192. 210. 223. 237. 235. 11000 229. 251. 264. 270. 274. 11001 207. 225. 238. 249. 255. 11002 209. 226. 239. 243. 256. 11003 197. 223. 238. 249. 249. 11004 212. 240. 254. 260. 270. 11005 207. 236. 253. 262. 263. 11006 211. 238. 246. 257. 270. 11007 208. 237. 240. 268. 275. 11008 210. 232. 241. 260. 275. 11600 213. 237. 261. 262. 264. 11601 247. 262. 280. 284. 290. 11602 216. 238. 226. 233. 234. 11603 229. 238. 253. 257. 258. 11604 206. 238. 242. 244. 257. 11605 228. 252. 268. 275. 284. 11606 219. 235. 240. 258. 254. 11607 221. 232. 240. 249. 249. 11608 211. 235. 251. 260. 256. _ _ _ _ 11609 ______ _ _ _ _ _ _ _ 2_ 2_ _9_. 240. 253. 272. 279. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 242. 246. 282. 281. 256. 263. 263. 252. 264. 269. 279. 287. 283. 286. 282. 277. 286. 288. 292. 280. 279. 288. 288. 305. 237. 240. 265. 275. 268. 277. 294. 299. 258. 269. 253. 264. 272. 287. 284. 288. __._ mvrocoi, 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 9): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DAY 1 8 15 22 29 RAT # DOSAGE GROUP 9 11009 195. 213. 222. 225. 228. 11010 216. 240. 250. 253. 268. 11011 217. 237. 253. 267. 275. 11012 203. 222. 233. 236. 234. 11013 199. 218. 225. 228. 232, 11014 222. 235. 254. 265. 267. 11015 220. 239. 252. 259. 269. 11016 212. 223. 229. 236. 240. 11017 199. 225. 248. 256. 260. 11018 217. 243. 256. 258. 276. 11610 205. 234. 248. 254. 243. 11611 232. 256. 265. 288. 290. 11612 219. 229. 230. 239. 244. 11613 228. 242. 260. 269. 276. 11614 210. 219. 234. 242. 248. 11615 233. 244. 253. 260. 270. 11616 218. 226. 240. 246. 242. 11617 226. 238. 244. 254. 262. 11618 208. 230. 240. 253. 261. 11619 226. 246. 254. 258. _ _ 2_ 5_ 2_ _. 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 56b 63 70 77 81 0.8 MG/KG PFOS 236. 235. 280. 258. 277. 288. 242. 242. 240. 240. 281. 287. 319. 322. 326. 316. 317. 322. 274. 277. 247. 234. 275. 278. 278. 264. 254. 255. 296. 298. 251. 255. 279. 289. 254. 266. 280. 282. 238. 252. 266. 276. 280. 285. 261. 259. . _ __ __ _ _ _ _ _ . _ _ _ _ . _ _ _ _ . . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ P c- 9"0 c- oo PROfOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 10): BODY WEIGHTS - PKEMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 22 29 36 42a 49 56b 63 70 77 81 .. - - - - - 1 MG/KG PFOS . . . . _ _ _ _ . _ . _ . _ _ . _ _ _ _ _ _ .__... ._.. 11019 196. 219. 228. 235. 243. 11020 213. 256. 267. 278. 281. 11021 208. 224. 230. 247. 258. 11022 214. 231. 241. 251. 270. 11023 208. 222. 231. 240. 242. 11024 210. 232. 240. 264. 275. 11025 211. 230. 233. 244. 249. 11026 211. 215. 240. 249. 267. 11027 198. 209. 216. 226. 234. 11028 217. 244. 259. 270. 272. 11620 206. 212. 221. 230. 237. 11621 244. 258. 264. 269. 273. 11622 223. 235. 232. 239. 244. 11623 221. 239. 247. 246, 253. 11624 211. 224. 232. 247. 249. 11625 231. 249. 258. 270. 272. 11626 216. 225. 243. 255. 256. 11627 220. 248. 266. 277. 292. 11628 206. 223. 240. 249. 260. 11629 220. 243. 252. 269. 272. .-_--..-___.----______ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 252. 292. 260. 278. 250. 285. 251. 281. 238. 281. 237. 273. 250. 258. 251. 288. 262. 303. 268. 281. ___. 90 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 26 (PAGE 11): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS . - - .- - - .. .- - - .- - - - - - 11029 203. 228. 227. 230. 248. 11030 214. 251. 264. 275. 272. 11031 206. 233. 247. 257. 273. 11032 216. 231. 250. 260. 265. 11033 206. 223. 235. 246. 254. 11034 212. 231. 241. 261. 268. 11035 209. 223. 233. 237. 249. 11036 213. 249. 255. 267. 267. 11037 195. 214. 219. 234. 236. 11038 212. 238. 247. 254. 258. 11630 202. 210. 224. 232. 231. 11631 237. 245. 263. 267. 271. 11632 221. 222. 239. 248. 252. 11633 223. 243. 255. 272. 275. 11634 210. 228. 243. 251. 269. 11635 239. 254. 257. 257. 273. 11636 219. 231. 231. 241. 249. 11637 223. 236. 239. 248. 250. 11638 208. 219. 217. 228. 231. . . 11639 _.__ _ _ 228. __ 253. ___ _ 2_ 5_ _ 7_ _. _ 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. 262. 282. 284. 265. 260. 278. 253. 276. 248. 263. 241. 275. 258. 281. 269. 278. 255. 256. 231. 272. ___- 238. 286. 316. 322. 318. 306. 305. 308. 272. 280. 263. 274. 254. 281. 237. 260. 248. 276. 255. 290. 275. 279. 262. 256. 233. 272. .__.______________________________ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 12): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11039 201. 229. 239. 239. 252. 11040 230. 241. 250. 258. 271. 11041 211. 239. 249. 256. 255. 11042 222. 241. 252. 266. 273. 11043 201. 224. 232. 234. 243. 11044 215. 232. 242. 252. 252. 11045 212. 230. 237. 245. 256. 11046 210. 241. 254. 261. 263. 11047 207. 223. 238. 242. 253. 11048 212. 230. 232. 238. 248. 11049 207. 231. 240. 248. 259. 11050 209. 245. 249. 256. 268. 11051 208. 230. 233. 241. 241. 11052 219. 231. 251. 261. 269. 11640 206. 212. 232. 234. 240. 11641 232. 262. 273. 280. 280. 11642 221. 241. 243. 253. 255. 11643 218. 233. 238. 244. 247. 11644 209. 220. 225. 229. 230. 11645 233. 252. 260. 276. 278. 11646 209. 230. 230. 236. 238. 11647 225. 243. 249. 256. 259. 11648 205. 218. 219. 225. 222. 11649 236. 259. 263. 279. 287. 11650 213. 228. 232. 229. 239. 11651 229. 250. 265. 272. 273. 11652 213. 232. 249. 254. 266. 11653 225. 236. 241. _ _ _ _ . _ _ _ __ _ _ _ _ _ . _ _ _ _ _ . 250. 253. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. Last value recorded before cohabitation. b. First value recorded after cohabitation. 260. 250. 283. 269. 265. 264. 265. 270. 273. 250. 244. 259. 255. 259. 252. 272. 261. 255. 259. 260. 247. 260. 254. 274. 270. 243. 246. 268. 267. 245. 252. 285. 291. 254. 262. 244. 245. 232. 237. 288. 290. 246. 256. 262. 265. 237. 238. 284. 292. 237. 239. 282. 280. 273. 274. 255. 256. . . . _ _._ _ . _ 288. 281. 277. 275. 272. P i b03 i 2PP 0 M w i PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 26 (PAGE 13): BODY WEIGHTS - PREMATING - INDIVIDUAL DATA - F O GENERATION FEMALE RATS DAY 1 .- - . ...- . _ 8 15 RAT # .- - .- .. ..- . 11053 11054 11055 11056 11057 11058 11059 11060 11061 11062 11063 11064 11065 11066 11654 11655 11656 11657 11658 11659 11660 11661 I1662 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. 218. 226. 246. 262. 280. 228. 234. 238. 225. 217. 221. 215. 213. 220. 241. 245. 262. 228. 233. 231. 232. 231. 253. 228. 241. 254. 230. 230. 240. 232. 245. 256. 235. 243. 250. 221. 235. 249. 237. 249. 254. 220. 226. 224. 243. 247. 243. 232. 244. 241. 244. 262. 269. 218. 237. 239. 235. 246. 256. 227. 240. 242. 227. 232. 240. 235. 243. 257. 243. 251. 260. 228. 237. 242. 239. 243. 252. 232. 239. 237. 242. 262. 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. 230. 284. 298. 245. 247. 232. 214. 231. 228. 259. 263. 253. 243. 262. 252. 266. 266. 247. 250. 261. 261. 264. 263. 295. 256. 258. 260. 253. 237. 242. 237. 247. 245. 250. 276. 282. 245. 250. 256. 259. 246. 252. 246. 256. 265. 273. 272. 272. 239. 242. 254. 253. 248. 253. 282. 287. _ _ _ _ _ _ . _ _ _ _ _ _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF STUDY a. L a s t value recorded before cohabitation. b. First value recorded after cohabitation. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 1): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS VEHICLE CONTROL ______-_________ ____ .- .- - - - - - . 10901 P 263. 264. 275. 277. 282. 284. 288. 289. 289. 294. 300. 304. 310. 10902 P 250. 258. 265. 262. 267. 268. 270. 277. 276. 277. 283. 294. 290. 10903 P 284. 293. 302. 298. 299. 300. 306. 308. 315. 316. 321. 323. 325. 10904 P 274. 283. 290. 296. 300. 295. 290. 299. 294. 304. 303. 316. 320. 10905 P 244, 264. 266. 269. 273. 279. 279. 288. 291. 293. 299. 303. 305. 10906 P 299. 305. 308. 303. 312. 313. 318. 320. 327. 324. 338. 332. 344. 10907 P 291. 298. 286. 306. 306. 310. 311. 314. 314. 326. 332. 339. 346. 10908 P 264. 276. 288. 283. 282. 290. 287. 290. 296. 304. 300. 311. 313. 10909 P 262. 273. 281. 285. 288. 291. 292. 294. 290. 293. 295. 302. 308. 10910 P 300. 312. 310. 317. 317. 318. 322. 323. 327. 331. 336. 341. 341. 10911 P 279. 279. 289. 293. 294. 296. 297. 302. 297. 301. 303. 307. 312. 10912 NP 289. 295. 302. 310. 310. 318. 321. 327. 338. 337. 344. 340. 330. 10913 P 279. 285. 290. 277. 281. 280. 292. 290. 297. 292. 302. 307. 308. 10914 P 250. 255. 261. 264. 266. 267. 268. 270. 267. 271. 278. 283. 285. 11501 P 232. 248. 242. 246. 252. 248. 250. 255. 251. 256. 260. 269. 271. 11502 P 306. 320. 330. 328. 324. 326. 325. 329. 331. 333. 337. 343. 348. 11503 P 274. 288. 289. 296. 295. 304. 305. 305. 305. 309. 315. 323. 327. 11504 P 286. 287. 293. 295. 295. 294. 300. 302. 307. 316. 318. 325. 328. 11505 P 274. 276. 279. 289. 287. 290. 289. 296. 292. 297. 305. 303. 316. 11506 P 302. 308. 316. 315. 322. 320. 322. 328. 329. 330. 338. 343. 353. 11507 P 266. 284. 290. 290. 292. 297. 296. 301. 305. 312. 311. 314. 323. 11508 P 287. 294. 294. 293. 297. 296. 299. 304. 307. 313. 319. 323. 325. 11509 P 272. 284. 294. 293. 294. 297. 298. 295. 299. 300. 303. 312. 319. 11510 P 310. 302. 306. 299. 307. 309. 315. 317. 316. 322. 320. 320. 328. 11511 P 248. 255. 256. 261. 267. 272. 268. 272. 269. 278. 285. 289. 292. 11512 P 306. 317. 319. 317. 320. 324. 328. 333. 335. 343. 344. 352. 353. P 11513 P 243. 251. 256. 254. 257. 256. 261. 263. 261. 263. 267. 273. 275. e-' 4Q 11514 P 281. 291. 298. 296. 301. 303. 300. 306. 303. 311. 315. 321. 326. 0 _ _ _ _ - - - - - _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - _- _ - - _ - _ _ _ _ _._ __ ._ _._ _ - _-_ - _ - - - - - -_ - - _--..- - - - - - - - - - C-L- - - . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION tn e-' P 00 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 2): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10901 P 10902 P 10903 P 10904 P 10905 P 10906 P 10907 P 10908 P 10909 P 10910 P 10911 P 10912 NP 10913 P 10914 P 11501 P 11502 P 11503 P 11504 P 11505 P 11506 P 11507 P 11508 P 11509 P 11510 P 11511 P 11512 P 11513 P 11514 P 312. 299. 330. 323. 312. 342. 342. 314. 309. 346. 312. 335. 320. 287. 276. 348. 331. 339. 313. 356. 333. 331. 326. 340. 291. 364. 283. 336. 319. 302. 340. 323. 321. 351. 342. 317. 308. 348. 320. 342. 320. 290. 281. 355. 341. 338. 319. 366. 333. 340. 332. 340. 298. 362. 283. 334. 324. 312. 347. 338. 326. 351. 356. 329. 319. 354. 330. 341. 328. 291. 282. 367. 337. 345. 324. 372. 339. 348. 331. 350. 307. 369. 294. 346. 334. 319. 359. 343. 337. 369. 361. 336. 321. 358. 341. 332. 336. 302. 295. 381. 344. 358. 332. 382. 348. 358. 337. 365. 312. 377. 299. 359. 340. 320. 367. 357. 350. 380. 377. 348. 331. 373. 358. 339. 354. 308. 302. 388. 365. 371. 345. 400. 369. 371. 352. 375. 321. 396. 312. 381. 352. 337. 388. 385. 362. 394. 379. 363. 349. 388. 374. 340. 370. 312. 320. 411. 366. 385. 355. 421. 374. 391. 361. 400. 331. 408. 323. 393. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 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. 4_ 0_ _4_. 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. 411. 386. 442. 411. 338. 418. 339. 348. 439. 406. 432. 403. 471. 407. 424. 389. 436. 376. 381. 447. 356. 416. __________...______.____ m c-' P W PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .-..____........_._....~~~~~~~.~ PREGNANCY STATUS DAY 0 1 ..___.__..______..._____________ RAT # DOSAGE GROUP 2 .. - _ _ _ . _ . _ _ _ _ _ . _ . _ . _ _ _ _ _ _ _ _ _ _ _ _ 10915 P 10916 P 10917 P 10918 P 10919 P 10920 P 10921 P 10922 P 10923 P 10924 P 10925 P 10926 P 10927 P 10928 P 11515 P 11516 P 11517 P 11518 P 11519 P 11520 P 11521 P 11522 NP 11523 P 11524 P 11525 P 11526 P 11527 P 11528 P 267. 297. 269. 281. 259. 288. 283. 288. 274. 274. 271. 280. 319. 246. 228. 296. 271. 308. 283. 268. 292. 293. 260. 248. 274. 275. 262. 272. 273. 299. 275. 293. 268. 293. 288. 290. 272. 278. 273. 295. 326. 257. 241. 311. 277. 315. 288. 276. 306. 290. 268. 256. 287. 276. 274. 287. 275. 303. 279. 291. 276. 291. 287. 298. 272. 284. 278. 301. 328. 261. 241. 315. 281. 323. 290. 275. 306. 298. 268. 264. 291. 284. 282. 287. 276. 303. 276. 291. 273. 300. 284. 294. 274. 289. 282. 301. 328. 264. 242. 311. 282. 322. 290. 274. 308. 305. 266. 263. 297. 287. 278. 294. 278. 304. 278. 290. 279. 306. 291. 295. 275. 287. 285. 305. 332. 268. 243. 312. 286. 323. 292. 275. 306. 297. 270. 264. 300. 285. 277. 296. 281. 309. 279. 291. 280. 303. 296. 299. 276. 292. 289. 304. 334. 274. 241. 308. 287. 324. 294. 272. 314. 295. 274. 264. 301. 288. 277. 300. 286. 314. 280 I 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. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 286. 313. 285. 300. 289. 307. 309. 302. 286. 296. 296. 308. 337. 279. 243. 310. 292. 324. 296. 289. 310. 302. 217. 270. 307. 291. 292. 297. 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. - - - - - - - 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. 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. ._________ 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. 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 24 25 _________.__..___...____________________----------.-------------- RAT # DOSAGE GROUP 2 MEVALONIC ACID CONTROL _.._____________________________________.---.------------.---- 10915 P 10916 P 314. 340. 316. 344. 328. 352. 335. 365. 335. 376. 355. 388. 370. 411. 377. 426. 402. 452. 10917 P 305. 308. 311. 322. 331. 346. 358. 366. 383. 10918 P 334. 343. 350. 362. 378. 386. 393. 420. 443. 10919 P 316. 320. 327. 335. 348. 345. 362. 375. 391. 10920 P 338. 342. 347. 364. 365. 379. 394. 412. 426. 10921 P 318. 318. 336. 344. 358. 372. 388. 398. 422. 10922 P 321. 330. 328. 335. 344. 356. 370. 385. 393. 10923 P 306. 308. 316. 325. 334. 340. 349. 362. 385. 10924 P 317. 325. 332. 341. 351. 359. 375. 381. 400. 10925 P 314. 323. 334. 341. 360. 376. 390. 405. 428. 10926 P MORIBUND SACRIFICED ON DAY 12 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 10927 P 348. 353. 363. 364. 385. 398. 412. 433. 10928 P 301. 300. 309. 318. 331. 351. 367. 386. 408. 11515 P 265. 273. 279. 286. 293. 304. 317. 332. 340. 11516 P 334. 350. 349 I 358. 375. 390. 407. 427. 436. 11517 P 319. 321. 331. 339. 352. 368. 374. 386. 393. 11518 P 343. 342. 354. 354. 373. 380. 404. 413. 422. 11519 P 319. 324. 329. 343. 354. 371. 387. 405. 406. 11520 P 309. 309. 322. 330. 348. 358. 366. 372. 386. 11521 P 336. 340. 341. 354. 357. 361. 375. 378. 387. 11522 NP 309. 306. 303. 305. 304. 306. 298. 295. 302. 304. 304. 302. 307. 11523 P 303. 306. 311. 324. 331. 340. 355. 376. 382. 11524 P 286. 294. 304. 311. 320. 339. 346. 355. 369. 11525 P 341. 341. 354. 360. 373. 388. 402. 418. 421. 11526 P 312. 318. 327. 334. 345. 358. 433. 384. 403. P + 11527 P 312. 316. 322. 332. 347. 363. 370. 384. 393. 11528 P 326. 330. 339. 349. 362. 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 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) 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 # DOSAGE GROUP 3 1.6 MG/KG PFOS ..- - - - - .- - - - - - - ..- _ . - - - - _ _ _ _. - _ - - -_ - - -_- _ - . - . - _ _ _ _ 10929 P 237. 241. 249. 250. 254. 255. 256. 10930 P 275. 281. 282. 284. 285. 280. 280. 10931 P 246. 253. 253. 252. 254. 257. 255. 10932 P 251. 254. 261. 262. 260. 266. 270. 10933 P 234. 241. 241. 242. 247. 246. 250. 10934 P 265. 272. 276. 279. 282. 281. 286. 10935 P 239. 240. 246. 246. 245. 245. 251. 10936 FOUND DEAD ON DAY 2 OF STUDY 10937 P 266. 271. 275. 272. 274. 274. 274. 10938 P 294. 290. 287. 285. 289. 287. 295. 10939 NP 250. 252. 254. 258. 257. 258. 260. 10940 P 284. 291. 294. 297. 297. 296. 302. 10941 P 244. 250. 247. 250. 253. 252. 252. 10942 P 282. 280. 279. 280. 280. 277. 281. 11529 P 252. 254. 249. 249. 250. 254. 250. 11530 P 313. 316. 319. 324. 320. 320. 327. 11531 NP 266. 251. 260. 260. 255. 252. 263. 11532 P 282. 283. 281. 284. 287. 293. 289. 11533 P 255. 264. 264. 259. 271. 269. 276. 11534 P 305. 306. 306. 303. 307. 308. 309. 11535 P 11536 P 268. 268. 269. 275. 274. 283. 274. 289. 275. 286. 278. 285. 278. 280. 11537 P 262. 265. 270. 270. 272. 270. 267. 11538 P 267. 274. 271. 271. 271. 271. 274. 11539 P 223. 232. 232. 235. 237. 235. 236. 11540 P 11541 P 298. 255. 301. 265. 306. 271. 308. 261. 307. 263. 308. 263. 315. 267. 11542 P 11543 P 282. 286. 284. 307. 283. 310. 280. 313. 286. 313. 292. 312. 288. 318. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION + MEVALONIC ACID __--- 264. 263. 285. 293. 260. 262. 269. 276. 249. 251. 287. 290. 247. 249. 278. 290. 257. 301. 256. 287. 258. 322. 260. 292. 271. 311. 280. 288. 272. 281. 236. 313. 273. 291. 320. ______ 283. 299. 253. 308. 261. 286. 260. 324. 257. 297. 268. 310. 280. 293. 271. 279. 239. 313. 269. 291. 325. __--- 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. 3_ 2_ _8_. 10 11 12 --__ 274. 295. 274. 283. 250. 293. 258. - -- -- 275. 298. 274. 287. 261. 296. 260. ______.___ 282. 304. 281. 290. 262. 301. 267. 289. 306. 254. 313. 266. 288. 273. 332. 261. 303. 279. 328. 289. 293. 281 ' 283. 244. 323. 277. 304. 330. 292. 312. 258. 325. 268. 296. 273. 341. 261. 309. 278. 331. 294. 300. 290. 285. 248. 329. 282. 311. 340. 300. 320. 252. 319. 269. 299. 275. 344. 260. 312. 284. 336. 302. 310. 290. 297. 247. 340. 289. 313. 341. I'KOTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 6): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 10929 P 282. 289. 300. 310. 308. 327. 342. 359. 381. 10930 P 308. 307. 321. 320. 338. 356. 365. 370. 386. 10931 P 280. 285. 288. 302. 315. 332. 347. 357. 382. 10932 P 292. 290. 308. 308. 324. 338. 356. 369. 401. 10933 P 264. 273. 289. 300. 318. 337. 350. 362. 392. 10934 P 304. 310. 316. 322. 332. 352. 360. 370. 391. 10935 P 273. 271. 287. 291. 301. 310. 332. 344. 363. 10936 FOUND DEAD ON DAY 2 OF STUDY 10937 P 294. 304. 309. 319. 330. 348. 350. 364. 383. 10938 P 320. 326. 343. 357. 367. 387. 401. 416. 436. 10939 NP 249. 253. 254. 258. 250. 255. 254. 258. 253. 254. 259. 256. 256. 10940 P 317. 331. 341. 351. 364. 387. 407. 441. 459. 10941 P 277. 278. 280. 298. 314. 333. 334. 343. 356. 10942 P 299. 309. 310. 329. 346. 359. 370. 382. 399. 11529 P 280. 291. 296. 302. 323. 335. 347. 354. 375. 11530 P 355. 359. 367. 375. 391. 416. 422. 443. 453. 11531 NP 262. 267. 267. 268. 268. 274. 270. 270. 263. 266. 268. 267. 267. 11532 P 323. 324. 331. 344. 358. 377. 390. 414. 11533 P 290. 298. 301. 315. 328. 345. 351. 370. 11534 P 346. 357. 353. 366. 384. 400. 416. 427. 11535 P 307. 310. 316. 327. 350. 361. 385. 397. 11536 P 310. 320. 316. 328. 343. 356. 360. 374. 11537 P 296. 297. 310. 320. 339. 354. 366. 379. 11538 P 301. 318. 319. 321. 336. 347. 362. 370. 11539 P 254. 258. 268. 269. 288. 301. 311. 325. 11540 P 345. 347. 354. 368. 389. 400. 418. 433. 11541 P 294. 292. 300. 309. 328. 338. 350. 363. 11542 P 321. 322. 326. 344. 355. 368. 385. 399. 11543 P 343. 353. 364. 373. 385. 402. 414. 426. ...---...------__-----~--~~~~-~~~~~.~-.~~~~~~~ ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 7): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS ..- - - ..- - 10943 P 10944 P 10945 P 10946 P 10947 P 10948 P 10949 P 10950 P 246. 275. 283. 276. 251. 287. 262. 268. 245. 273. 294. 279. 251. 285. 255. 269. 255. 271. 295. 273. 253. 291. 254. 275. 256. 267. 294. 283. 254. 296. 256. 278. 256. 265. 292. 278. 255. 297. 262. 282. 259. 264. 294. 275. 258. 297. 266. 288. 262. 267. 294. 282. 258. 302. 266. 288. 264. 276. 296. 280. 259. 302. 267. 293. 262. 272. 298. 283. 267. 313. 265. 297. 266. 275. 301. 293. 264. 314. 274. 298. 274. 279. 306. 290. 270. 322. 276. 306. 281. 283. 313. 300. 278. 328. 288. 310. 284. 284. 316. 304. 273. 325. 290. 312. 10951 P 10952 P 284. 285. 280. 287. 282. 288. 275. 287. 269. 290. 280. 288. 279. 292. 284. 294. 283. 295. 286. 293. 295. 289. 296. 299. 298. 300. 10953 P 249. 258. 254. 246. 252. 252. 253. 254. 255. 262. 268. 271. 275. 10954 P 10955 P 270. 251. 272. 254. 276. 257. 278. 259. 279. 254. 283. 258. 280. 260. 284. 258. 285. 266. 288. 264. 290. 273. 299. 272. 293. 274. 10956 P 11544 P 11545 P 276. 261. 312. 291. 265. 321. 282. 267. 331. 284. 271. 337. 281. 268. 336. 285. 269. 341. 288. 269. 342. 290. 268. 344. 298. 269. 339. 301. 269. 348. 306. 274. 349. 310. 279. 357. 309. 287. 362. 11546 P 11547 P 11548 NP 11549 P 11550 P 11551 P 11552 P 11553 P 11554 P 260. 284. 254. 277. 250. 280. 276. 278. 260. 267. 288. 257. 281. 258. 287. 280. 280. 264. 274. 290. 255. 288. 260. 291. 283. 282. 263. 270. 291. 261. 288. 262. 292. 282. 281. 264. 272. 291. 261. 287. 263. 292. 282. 282. 274. 273. 290. 269. 293. 265. 291. 285. 287. 277. 277. 292. 265. 291. 267. 294. 287. 285. 266. 270. 299. 270. 297. 265. 304. 287. 281. 277. 276. 297. 269. 296. 269. 301. 292. 287. 277. 280. 309. 261. 301. 268. 304. 291. 291. 275. 288. 308. 260. 308. 277. 312. 298. 293. 277. 288. 319. 260. 313. 281. 318. 300. 299. 287. 292. 326. 259. 313. 285. 321. 307. 304. 292. 11555 P 11556 P 11557 P 293. 250. 300. 292. 262. 305. 302. 266. 307. 301. 269. 300. 299. 273. 304. 299. 274. 305. 300. 275. 310. 308. 274. 3_ 1_ _1_. 307. 316. 279. 281. 310. 317. __.. _ _ _ _ _ 316. 286. 324. 322. 294. 3_ 3_ 3_ ._ 332. 298. 334. .- - - - - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). 0 P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ? M PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS [SPONSOR*SSTUDY 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 22 23 24 25 __...____...____________________________---.------.----------.-.--- RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 P 10944 P 10545 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 1.1557 P 292. 296. 300. 312. 330. 346. 360. 371. 382. 294. 297. 301. 314. 327. 347. 354. 370. 323. 325. 332, 344. 363. 376. 396. 401. 415. 303. 303. 313. 325. 341. 360. 385. 401. 421. 280. 284. 292. 302. 312. 332. 349. 360. 385. 331. 338. 341. 358. 367. 391. 405. 423. 437. 295. 269. 253. 249. 260. 267. 276. 278. 280. 313. 319. 324. 334. 349. 368. 380. 398. 421. 303. 308. 322. 335. 344. 364. 376. 385. 417. 304. 304. 316. 326. 339. 356. 363. 374. 277. 284. 296. 307. 326. 340. 347. 364. 399. 308. 303. 315. 319. 333. 350. 369. 384. 407. 286. 292. 258. 308. 327. 341. 358. 376, 319. 324. 329. 339. 355. 368. 379. 394. 296. 304. 310. 322. 335. 356. 372. 391. 399. 370. 372. 378. 386. 400. 414. 423. 438. 449. 302. 305. 309. 320. 337. 347. 356. 373. 380. 328. 340. 339. 351, 367. 380. 387. 400. 418. 261. 261. 260. 256. 258. 254. 258. 262. 259. 258. 262. 263. 272 316. 322. 332. 344. 358. 373. 386. 401. 290. 298. 304. 310. 324. 330. 349. 369. 329. 331. 344. 359. 377. 388. 403. 412. 423. 312. 314. 318. 331. 346. 359. 371. 392. 403. 308. 316. 323. 339. 355. 374. 386. 392. 293. 301. 311. 324, 343. 353. 371. 372. 336. 336. 344. 351. 360. 385. 397. 415. 301. 308. 320. 337. 343. 361. 378. 383. 336. 338. 359. 362. 376. 388. 378. 386. ._____ ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION td PROTOCOL 418-01.8: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 9 ) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 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. 317. 280. 276. 248. 323. 298. 311. 264. 303. 307. 274. 276. 304. 257. 292, 310. 304. MORIBUND 295. 276. 260. 284. 276. 252. 313. 254. 294. 265. 274. 323. 330. 294. 296. 285. 292. 257. 267. 332. 333. 307. 306. 317. 316. 265. 271. 308. 316. 310. 314. 280. 284. 282. 288. 314. 318. 263. 272. 301. 309. 304. 306. 313. 314. SACRIFICED ON DAY 311. 314. 282. 296. 271. 272. 289. 286. 289. 295. 261. 262. 315. 317. 262. 266. 309. 318. 276. 324. 298. 299. 265. 332. 312. 321. 274. 318. 314. 285. 289. 308. 263. 319. 308. 317. 16 OF 320. 296. 275. 292. 299. 269. 324. 263. 326. 283. 322. 304. 298. 274. 328. 319. 319. 278. 320. 320. 285. 291. 319. 264. 318. 303. 321. STUDY 320. 299. 280. 296. 301. 270. 331. 270. 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. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 250. 330. 307. 306. 276. 337. 328. 325. 282. 330. 328. 292. 297. 323. 268. 325. 303. 329. 263. 334. 311. 311. 280. 333. 332. 324. 282. 331. 326. 294. 298. 323. 275. 327. 307. 338. 267. 340. 316. 314. 286. 337. 342. 330. 288. 336. 330. 299. 301. 327. 278. 330. 306. 343. 327. 329. 339. 300. 300. 308. 287. 292. 294. 306. 305. 308. 310. 316. 320. 277. 277, 284. 333. 336. 344. 269. 276. 277. 329. 338. 348. .____ .. .- - - - .. 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. 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. ____ 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. 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. .- - - - - - PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AM) NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 7 (PAGE 10): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 10957 P 302. 303. 312. 314. 328. 340. 349. 361. 378. 10958 P 352. 351. 361. 368. 381. 398. 415. 426. 448. 10959 P 335. 337. 332. 358. 370. 394. 412. 427. 448. 10960 P 338. 345. 356. 368. 380. 390. 407. 417. 442. 10961 P 300. 298. 312. 318. 341. 359. 373. 385. 409. 10962 P 360. 358. 369. 383. 393. 403. 419. 422. 444 I 10963 P 358. 366. 375. 394. 406. 420. 436. 456. 475. 10964 P 357. 356. 366. 382. 401. 410. 435. 452. 476. 10965 P 294. 303. 316, 325. 337, 344. 353. 369. 386. 10966 P 359. 364. 372. 376. 387. 392. 407. 424. 434. 10967 P 347. 353. 357. 364. 377 I 394. 405. 419. 430. 436. 10968 P 316. 323. 334, 340. 353. 366. 381. 399. 10969 P 319. 324. 328. 338. 348. 364. 384. 397. 413. 10970 P 349. 348. 358. 368. 378. 388. 398. 413. 429. 11558 P 295. 308. 315. 322. 343. 351. 366. 371. 383. 11559 P 359. 367. 376. 388. 405. 419. 433. 456. 462. 1 1 5 6 0 NP 304. 311. 309. 308. 307. 309. 306. 312. 312. 314. 314. 11561 P 353. 363. 366. 370. 386. 405. 402. 415. 434. 11562 MORIBUND SACRIFICED ON DAY 16 OF STUDY 11563 P 365. 368. 365. 375. 379. 397. 413. 425. 448. 11564 P 325. 339. 342. 346. 355 I 371. 381. 395. 405. 405. 11565 P 321. 325. 336. 349. 360. 370. 381. 397. 402. 1 1 5 6 6 NP 299. 299. 294. 290. 297, 292. 287. 290. 291. 296. 294. 11567 P 343. 351. 355. 369. 378, 389. 408. 415. 438. 11568 P 304. 315. 321. 322. 339. 352. 366. 373. 393. 11569 P 368. 372. 377 I 393. 409, 425. 441. 445. 466. 11570 P 298. 303. 305. 312. 320. 329. 339. 355. 372. 11571 P 375. 378. 383. 391. 400. 410. 423. 444. 457. . _ _ _ _ _ _ _ _ . . _ _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ .- -_ - ._ - - -_ _ __ __ __ . _ _ - - . - . - . - - - . . . - - - . - - - - - - - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 11): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS RAT # DOSAGE GROUP 6 1.6 MG/KG_ P_ F_ O_ S_ f CHOLESTEROL ._._ ____ . _ _ - -_ - - -_ - - - _ _ _ _ _ _ _ _ _ _ 10971 P 231. 232. 236. 242. 245. 248. 248. 251. 248. 256. 261. 268. 271. 10972 P 287. 294. 293. 299. 300. 306. 306. 308. 319. 318. 320. 330. 333. 10973 P 239. 246. 249. 250. 255. 258. 258. 261. 263. 267. 273. 272. 278. 10974 P 267. 278. 280. 279. 282. 286. 291. 293. 301. 300. 311. 310. 315. 10975 P 267. 256. 269. 273. 269. 274. 279. 280. 282. 283. 285. 281. 282. 10976 P 272. 277. 280. 276. 274. 274. 274. 277. 279. 279. 289. 283. 283. 10977 P 254. 256. 256. 260. 257. 260. 263. 263. 267. 265. 271. 274. 284. 10978 P 274. 287. 284. 279. 283. 278. 290. 282. 287. 292. 296. 300. 301. 10979 P 270. 274. 282. 284. 291. 290. 294. 297. 297. 305. 310. 314. 320. 10980 P 260. 264. 265. 263. 265. 270. 272. 274. 266. 280. 282. 286. 290. 10981 P 250. 248. 251. 247. 253. 253. 253. 255. 257. 263. 267. 261. 266. 10982 P 266. 251. 259. 260. 264. 266. 268. 268. 272. 274. 281. 281. 286. 10983 P 263. 272. 272. 266. 272. 273. 278. 278. 282. 285. 296. 297. 303. 10984 P 11572 P 278. 210. 281. 219. 287. 223. 284. 223. 289. 226. 293. 221. 294. 228. 294. 232. 297. 231. 305. 239. 306. 242. 315. 246. 316. 246. 11573 P 278. 281. 287. 287. 289. 294. 293. 300. 297. 296. 306. 308. 318. 11574 P 263. 268. 266. 266. 268. 273. 272. 279. 279. 280. 285. 288. 293. 11575 P 274. 268. 269. 285. 291. 292. 294. 302. 304. 308. 313. 316. 327. 11576 P 276. 273. 272. 277. 273. 277. 282. 274. 289. 291. 293. 295. 298. 11577 P 286. 291. 297. 294. 295. 297. 306. 314. 309. 312. 321. 326. 329. 11578 P 248. 256. 262. 262. 264. 268. 270. 271. 272. 281. 285. 288. 294. 11579 P 298. 314. 314. 311. 314. 319. 320. 323. 323. 327. 336. 338. 348. 11580 P 275, 275. 282. 283. 283. 289. 292. 299. 304. 302. 309. 312. 327. 11581 P 278. 280. 284. 291. 288. 291. 292. 296. 297. 301. 302. 310. 306. 11582 P 262. 269. 275. 273. 276. 280. 286. 289. 286. 292. 293. 305. 306. rf: 11583 P 11584 P 266. 274. 264. 282. 268. 282. 269. 286. 268. 284. 281. 289. 278. 287. 274. 291. 279. 296. 292. 292. 295. 300. 296. 300. 301. 311. 11585 P 257, 254. 259. 264. 260. 262. 262. 258. .- - - - 265. 274. ____ 270. 284. 290. .____ .- .- - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 12): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS _____________._.__.....-.....--..-.-------------------...-..---.---------------- PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 22 23 24 25 RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL 10971 P 270. 274. 274. 280. 297. 304. 309. 321. 10972 P 10973 P 337. 282. 342. 287. 352. 287. 364. 296. 375. 311. 400. 322. 417. 332. 433. 343. 10974 P 325. 330. 342. 357. 374. 388. 407. 422. 10975 P 291. 291. 297. 302. 304. 319. 326. 333. 10976 P 288. 290. 299. 306. 322. 333. 348. 364. 10977 P 293. 292. 296. 304. 319. 335. 352. 360. 10978 P 303. 306. 323. 327. 336. 360. 367. 392. 10979 P 321. 327. 338. 344. 350. 370. 386. 400. 10980 P 292. 296. 313. 330. 348. 345. 355. 374. 10981 P 275. 274. 288. 292. 307. 316. 326. 344. 10982 P 289. 298. 310. 324. 343. 362. 376. 392. 10983 P 302. 304. 313. 331. 338. 358. 376. 391. 10984 P 320. 325. 334. 346. 358. 366. 383. 400. 11572 P 252. 254. 262. 270. 282. 294. 298. 312. 322. 11573 P 322. 325. 339. 342. 357. 375. 385. 399. 396. 11574 P 296. 304. 312. 327. 344. 352. 370. 374. 389. 11575 P 331. 336. 349. 353. 368. 387. 400. 411. 412. 11576 P 303. 312. 319. 324. 335. 352. 356. 373. 384. 11577 P 335. 347. 356. 370. 387. 414. 424. 434. 11578 P 297. 305. 306. 322. 337. 361. 375. 386. 396. 11579 P 342. 357. 359. 376. 392. 401. 413. 430. 11580 P 334 I 335. 346. 360. 373. 387. 409. 423. 436. 11581 P 315. 319. 321. 334. 350. 364. 377. 395. 398. 11582 P 318. 322. 329. 341. 344. 365. 372, 376. 397. 11583 P 315. 322. 323. 332. 354. 363. 381. 404. 11584 P 314. 312. 328. 341. 347. 362. 377. 383. _ 11585 P ____ _ 295 __ . _ _ 298. __ _ 301. ___ _ 31 _ 8. _ _ 3 _ 36. __ _ 346. __ _ 367. _ . __ __ _ _ _ _ 3.__ 7_ 3_ _._ _ _ _ _ _ _ _ . . ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 7 (PAGE 1 3 ) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10985 P 257. 269. 269. 271. 272. 10986 P 280. 277. 283. 278. 276. 10987 P 272. 283. 278. 278. 283. 10988 P 271. 280. 278. 273. 275. 10989 P 226. 230. 240. 248. 243. 10990 P 288. 289. 292. 290. 291. 10991 P 262. 265. 264. 264. 266. 10992 P 249. 246. 248. 255. 253. 10993 P 282. 266. 267. 283. 280. 10994 P 280. 274. 278. 276. 284. 10995 P 260. 269. 269. 268. 273. 10996 P 258. 268. 267. 267. 266. 10997 P 242. 244. 249. 247. 246. 1 0 9 9 8 NP 280. 275. 272. 278. 277. 11586 P 246. 249. 255. 252. 253. 11587 P 298. 296. 310. 304. 304. 11588 P 259. 259. 266. 267. 267. 11589 P 269. 274. 284. 275. 279. 11590 P 250. 258. 264. 263. 261. 11591 P 293. 301. 301. 296. 308. 11592 P 288. 296. 292. 293. 298. 11593 P 235. 228. 234. 242. 240. 11594 P 271. 277. 286. 284. 285. 11595 P 256. 262. 268. 269. 271. 11596 P 252. 258. 257. 255. 256. 11597 P 253. 257. 258. 258. 260. 11598 P 256. 261. 270. 270. 269. 11599 P 276. 276. 275. 268. 269. .. - - - - - - ....- - - - - - - - - .. - - - - - - _ _ _ . . . _ _ _ _ _ _ _ 272. 282. 279. 279. 252. 298. 271. 251. 285. 281. 271. 268. 252. 279. 254, 312. 275. 277. 264. 307. 296. 244. 275. 275. 259. 261. 272. 270. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 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. 279. 294. 299. 286. 289. 281. 280. 263. 264. 306. 309. 269. 270. 264. 260. 301. 300. 284. 290. 282. 286. 278. 275. 258. 262. 279. 270. 268. 276. 316. 326. 275. 279. 303. 300. 273. 280. 322. 324. 302. 297. 249. 260. 286. 290. 280. 284. 271. 271. 267. 276. 280 I 282. 284. 284. .____ 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. 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. 2_ 9_ 6_ _. 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. so 2 9 5 . 300. 0 __.___ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 22 23 24 25 _______.._______________________________.-------.----------.-.- RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL __..___..._.____________________________--------.---.-.---------- 10985 P 10986 P 10987 P 10988 P 10989 P 10990 P 10991 P 10992 P 10993 P 10994 P 10995 P 10996 P 10997 P 10998 NP 11586 P 11587 P 11588 P 11589 P 11590 P 11591 P 11592 P 11593 P 11594 P 11595 P 11596 P 11597 P 11598 P 11599 P 290. 311. 302. 293. 270. 323. 284. 270. 328. 303. 301. 284. 273. 271. 293. 348. 301. 315. 306. 356. 307. 274. 321. 305. 286. 284. 295. 300. 298. 316. 308. 307. 274. 328. 289. 286. 328. 298. 309. 290. 274. 262. 296. 353. 305. 323. 312. 360. 314. 275. 317. 298. 293. 288. 299. 307. 304. 324. 309. 318. 272. 340. 295. 292. 337. 317. 314. 307. 290. 264. 311. 366. 314. 329. 318. 368. 324. 284. 328. 303. 306. 290. 313. 317. 314. 348. 324. 325. 278. 345. 302. 302. 345. 330. 323. 314. 297. 269. 320. 376. 319. 346. 336. 377. 336. 288. 341. 314. 320. 303. 328. 334. 331. 364. 331. 332. 282. 359. 321. 316. 362. 344. 340. 324. 314. 269. 333. 394. 337. 364. 345. 389. 348. 311. 360. 323. 338. 316. 343. 350. 338. 371. 351. 355. 294. 366. 334. 325. 372. 364. 366. 347. 330. 269. 349. 404. 352. 380. 363. 411. 366. 322. 378. 332. 355. 331. 359. 364. 359. 396. 366. 368. 308. 378. 347. 347. 394. 382. 379. 354. 346. 266. 361. 423. 359. 397, 377. 425. 381. 337. 396. 342. 364. 341. 365. 379. 366. 416. 373. 374. 312. 387. 361. 360. 404. 394. 386. 365. 348. 265. 368. 432. 375. 410. 389. 446. 401. 354. 408. 357. 375. 355. 376. 392. 402. 431. 394. 397. 323. 403. 379. 372. 430. 424. 396. 364. 272. 375. 423. 418. 421. 360. 368. 416. 277. 366. 275. 274. 283. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODIJCTIONSTUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 15): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 11 12 RAT # DOSAGE GROUP 8 0.4 MG/KG PFOS ---- -------. -_________________________ 10999 P 250. 259. 265. 267. 267. 276. 280. 282. 288. 291. 297. 303. 307. 11000 P 298. 300. 302. 306. 308. 316. 317. 323. 328. 334. 331. 338. 344. 11001 P 273. 279. 280. 280. 281. 282. 283. 281. 292. 296. 298. 303. 304. 11002 P 281. 281. 283. 288. 287. 296. 295. 304. 303. 304. 314. 307. 314. 11003 P 284. 294. 296. 300. 301. 306. 304. 304. 310. 312. 307. 312. 323. 11004 P 281. 292. 299. 300. 307. 314. 315. 316. 315. 313. 324. 337. 340. 11005 P 291. 298. 301. 307. 307. 313. 318. 318. 325. 328. 327. 330. 337. 11006 P 273. 290. 290. 291. 300. 306. 302. 308. 312. 313. 319. 321. 325. 11007 P 298. 305. 319. 320. 330. 330. 334. 336. 339. 343. 348. 352. 357. 11008 P 301. 317. 315. 315. 318. 316. 323. 322. 316. 317. 328. 332. 333. 11600 P 284. 297. 306. 303. 304. 310. 319. 315. 319. 320. 328. 334. 341. 11601 P 300. 325. 324. 327. 328. 336. 336. 331. 336. 344. 350. 355. 363. 11602 P 242. 254. 252. 259. 262. 264. 267. 268. 273. 279. 281. 285. 292. 11603 P 282. 291. 298. 299. 301. 305. 309. 314. 314. 318. 322. 329. 332. 11604 P 278. 278. 276. 283. 285. 288. 290. 294. 299. 295. 301. 306. 320. 11605 P 305. 314. 319. 317. 323. 330. 329. 330. 337. 344. 341. 346. 352. 11606 P 274. 279. 280. 279. 285. 288. 291. 293. 295. 300. 298. 309. 312. 11607 P 256. 267. 270. 270. 272. 275. 275. 283. 280. 289. 292. 303. 310. 11608 P 291. 305. 314. 306. 315. 316. 322. 324. 327. 326. 330. 342. 344. 11609 P 289. 300. 303. 308. 311. 314. 316. 322. 327. 331. 338. 339. 345. . - _ _ _ _ _ - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ __ __ . _ _ _ _ _ __ _ _- _ _ -_ _ . -_ _ - - _ _ -. _ -_ -.- _ -_ - -_ - - _-_ - - _ -_ - -_ - - _ ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 1 6 ) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10999 P 310. 316. 327. 331. 339. 359. 368. 379. 11000 P 351. 351. 361. 367. 380. 397. 399. 422. 11001 P 306. 312. 323. 336. 349. 368. 384. 392. 11002 P 316. 327. 337. 349. 361. 369. 386. 403. 11003 P 322. 320. 348. 372. 380. 398. 402. 416. 11004 P 345. 347. 358. 365. 375. 390. 406. 422. 11005 P 338. 338. 343. 349. 364. 376. 385. 397. 412. 11006 P 330. 343. 348. 354. 368. 379. 395. 417. 430. 11007 P 355. 360. 377. 392. 405. 427. 428. 444. 459. 11008 P 331. 329. 341. 356. 373. 378. 401. 403. 423. 11600 P 343. 349. 357. 367. 381. 397. 403. 423. 438. 11601 P 366. 376. 385. 400. 412. 424. 435. 449. 469. 11602 P 294. 296. 303. 307. 325. 332. 336. 352. 350. 11603 P 338. 339. 351. 362. 374. 388. 409. 430. 11604 P 313. 321. 326. 333. 345. 363. 374. 386. 11605 P 357. 364. 370. 379. 393. 405. 426. 442. 11606 P 321. 321. 325. 332. 345. 352. 366. 380. 11607 P 306. 318. 330. 346. 354. 370. 380. 386. 11608 P 348. 363. 363. 364. 387. 390. 405. 419. 11609 P 350. 361. 369. 379. 398. 411. 428. 444. _ _ _ _ _ _ _ _ - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ __ __ __ __ __ . _ _ _ _ _ _ - - - - - - - - - - . - - - - - - - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 17): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11009 P 238. 252. 258. 255. 254. 253. 258. 259. 267. 268. 268. 11010 P 288. 296. 299. 307. 305. 303. 306. 311. 309. 321. 333. 11011 P 293. 296. 310. 308. 312. 313. 310. 313. 322. 319. 307. 11012 P 252. 258. 265. 266. 262. 262. 265. 269. 271. 279. 279. 11013 P 251. 251. 256. 259. 264. 266. 269. 271. 270. 277. 283. 11014 NP MATING NOT CONFIRMED 11015 P 296. 296. 298. 301. 293. 326. 290. 294. 301. 304. 306. 11016 P 236. 252. 253. 253. 256. 261. 268. 267. 266. 268. 271. 11017 NP 284. 290. 298. 296. 307. 308. 303. 307. 316. 319. 319. 11018 P 296. 308. 312. 307. 309. 308. 313. 310. 311. 319. 318. 11610 NP 248. 248. 251. 256. 260. 283. 272. 279. 279. 280. 284. 11611 P 304. 325. 318. 316. 323. 326. 324. 329. 327. 332. 344. 11612 P 258. 264. 269. 276. 270. 274. 278. 283. 283. 286. 289. 11613 P 283. 295. 303. 303. 306. 309. 315. 315. 327. 324. 333. 11614 P 277. 278. 277. 285. 286. 293. 285. 283. 296. 294. 304. 11615 P 293. 300. 301. 300. 304. 296. 304. 304. 306. 312. 316. 11616 P 254. 258. 256. 252. 261. 271. 263. 268. 274. 273. 286. 11617 P 275. 280. 281. 279. 286. 285. 285. 293. 297. 306. 310. 11618 P 296. 305. 312. 306. 310. 314. 319. 321. 316. 322. 326. 11619 P 268. 265. 267. 267. 271. 277, 275. 282. 275. 277. 286. _ _ _ _ . . . - - - - - - . _ _ _ _ _ _ _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ . . . . - - - . - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 272. 336. 332. 288. 282. 314. 282. 320. 322. 290. 347. 295. 334. 305. 322. 285. 310. 331. 290. 272. 345. 338. 291. 289. 312. 282. 321. 326. 289. 349. 301. 341. 309. 329. 287. 314. 340. 295. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 18); MATERNAL BODY WEIGHTS - PRESUMED GESTATION - TNDIVIDUAL DATA - Fo GENERATION FEMALE WTS . _ _ _ _ _ _.._ __ _._ __ __ . ._ _._ . . _ _ _ _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - - - - - - - . - - PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 22 23 24 25 _._...._._ ........................................................................................................................ . . RAT # _-- - _ DOSAGE __. GROUP ..- 9 _ _ _ _ _ _ _ . - - ~ _ _ _ _ 0.8 MG/KG PFOS ~~~~~ ~ . . . . - - ~ ~ ~ ~ - ~ ~ ~ ~ ~ ~ . . . - - . . . ~ ~ 11009 P 274. 273. 284. 301. 309. 328. 11010 P 346. 353. 361. 377. 393. 403. 11011 P 342. 349. 356. 370. 384. 398. 11012 P 293. 295. 312, 320. 335. 344. 11013 P 289. 291. 302. 313. 325. 337. 11014 NP MATING NOT CONFIRMED 11015 P 316. 326. 333. 342. 356. 371. 388. 406. 418. 11016 P 286. 288. 296. 302. 322. 339. 349. 364. 11017 NP 310. 309. 307. 303. 297. 302. 298. 288. 288. 289. 296. 294. 298. 11018 P 332. 338. 347. 356. 358. 368. 377. 394. 411. 339. 11610 NP 278. 280. 283. 293. 298. 300. 299. 299. 306. 306. 312. 308. 297. 11611 P 362. 364. 371. 378. 399. 416. 424. 435. 11612 P 304. 310. 316. 328. 335. 354. 367. 376. 397. 11613 P 344. 357. 364. 377. 388. 403. 416. 430. 449. 11614 P 315. 330. 338. 350. 366. 388. 403. 409. 414. 11615 P 333. 340. 339. 357. 365. 383 I 395. 408. 433. 11616 P 301. 301. 324. 330. 348. 359. 370. 380. 11617 P 321. 325. 334. 346. 365. 379. 396. 418. 11618 P 340. 348. 356. 365. 378. 385. 392. 405. 420. 11619 P 295. 310. 318. 324. 344. 356. 374. 387. 391. . _ . . . _ _ _ _ _ . _ - _ _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ . _ ___ __ _ _ _ __ _ __ _ _ _ _ _ ._ _ __ _-_ - - . _ - -_ - ..-_ - - _ -- - _ - -_ . -_ - - _ . _ - ---- -- - - - - - - - - ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 19): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS RAT # DOSAGE GROUP 10 1 MG/KG PFOS _ . - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - _ _ _ _ _ __ _ _ _ _ _ . _ ___ __ __ _ _ _ _ _._ -_ - ._ -_ - _ - _ - _ -_ _ - -_ -__ - .-- - --- -- - .-.-- - .- - -- - - --.--- - . - - - - - - - - - - 11019 P 277. 278. 279. 282. 283. 284. 287. 291. 285. 292. 297. 298. 301. 11020 P 296. 309. 315. 311. 308. 309. 313. 314. 319. 319. 320. 328. 333. 11021 P 254. 265. 269. 273. 278. 280. 280. 288. 289. 292. 293. 302. 301. 11022 NP 11023 P 304. 259. 304. 268. 304. 270. 308. 268. 312. 274. 316. 273. 312. 278. 318. 277. 318. 282. 316. 284. 318. 290. 317. 294. 310. 296. 11024 P 285. 300. 317. 314. 297. 323. 317. 322. 308. 320. 321. 328. 329. 11025 P 260. 265. 269. 273. 276. 277. 282. 281. 285. 289. 295. 298. 295. 11026 P 11027 P 289. 236. 295. 245. 296. 249. 301. 253. 300. 257. 302. 261. 302. 260. 301. 253. 303. 272. 310. 272. 314. 274. 319. 282. 320. 286. 11028 P 289. 302. 305. 303. 304. 304. 297. 307. 306. 307. 311. 319. 323. 11620 P 252. 259. 259. 263. 267. 266. 264. 266. 269. 269. 271. 275. 280. 11621 P 280. 290. 298. 300. 300. 299. 301. 305. 305. 307. 317. 323. 321. 11622 P 254. 261. 267. 269. 274. 275. 278. 282. 284. 290. 291. 299. 297. 11623 P 11624 P 275. 262. 276. 272. 280. 277. 278. 280. 271. 282. 284. 285. 290. 278. 292. 279. 291. 287. 295. 288. 305. 297. 302. 313. 306. 316. 11625 P 289. 306. 306. 307. 317. 311, 314. 314. 322. 322. 322. 328. 343. 11626 P 275. 275. 276. 279. 280. 285. 284. 287. 293. 298. 300. 310. 311. 11627 P 320. 326. 333. 324. 323. 327. 329. 334. 326. 331. 339. 346. 347. 11628 P 266. 275. 282. 282. 280. 282. 287. 289. 298. 304. 302. 301. 305. 11629 P 292. 303. 299. 304. 305. 305. 309. 309. 315. 314. 316. 326. 325. . - _ _ _ _ _ _ _ . _ . _ _ _ _ _ _ - . _ _ _ _ _ _ _ _ _ _ . _ _ _ r _ _ _ __ _ _ _ _ _ r _ _ _. - - - - - _ _ _ ___ __ _ _ __-_ -..-_ -_ - - -_- - -_--- --- - . - - ..- - - -- - --- - - - - - - - - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 20): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 11019 P 310. 313. 320. 331. 346. 365. 380. 398. 11020 P 334. 333. 347. 357. 366. 384. 396. 419. 433. 11021 P 308. 316. 322. 338. 358. 372. 387. 402. 412. 11022 NP 307. 309. 306. 302. 299. 299. 300. 303. 304. 310. 11023 P 302. 310. 324. 336. 352. 370. 378. 386. 11024 P 338. 344. 354. 358. 373. 379. 388. 404. 420. 11025 P 11026 P 11027 P 306. 324. 290. 310. 332. 300. 310. 332. 306. 330. 345. 316. 333. 358. 331. 349. 374. 343. 364. 386. 360. 381. 402. 377. 399. 411. 381. 403. 11028 P 11620 P 332. 286. 339. 293. 351. 304. 361. 317. 378. 319. 400. 342. 414. 355. 436. 369. 11621 P 328. 334. 341. 350. 369. 385. 405. 421. 11622 P 11623 P 11624 P 11625 P 11626 P 11627 P 11628 P 308. 316. 315. 343. 310. 350. 319. 317. 324. 325. 352. 326. 366. 330. 327. 323. 330. 360. 328. 372. 333. 334. 344. 341. 374. 339. 380. 346. 349. 359. 354. 388. 353. 389. 366. 364. 364. 369. 408. 364. 408. 376. 382. 385. 390. 423. 372. 391. 397. 396. 393. 404. 444. 377. 381. 414. 402. 412. 454. 395. 370. 11629 P 329. 341. 345. 354. 360. 378. 394. 400. 414. . . - - - _ _ _ _ _ _ _ . _ . . _ _ . _ _ - - -_ - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ __ _ __ _ __ _ __ _ __ _ __ _ _ __ __ __ __ __-_ _ _ _ _ . _ . _ _ _ _ _ . _ . - - - - - . ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 21): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11029 P 262. 274. 270. 253. 259. 277 275. 276. 276. 282. 283. 11030 NP MATING NOT CONFIRMED 11031 P 291. 295. 296. 302. 298. 304. 310. 311. 314. 314. 316. 11032 P 277. 283. 288. 288. 290. 291. 293. 294. 297. 297. 305. 11033 P 272. 272. 280. 274. 278. 275. 279. 280. 286. 285. 286. 11034 P 281. 286. 287. 286. 283. 284. 288. 292. 293. 296. 301. 11035 P 259. 268. 262. 266. 270. 278. 278. 276. 280. 272. 275. 11036 P 296. 296. 298. 297. 295. 303. 305. 314. 315. 319. 323. 11037 P 233. 243. 249. 247. 248. 250. 256. 255. 260. 266. 269. 11038 P 268. 277. 276. 280. 284. 282. 288. 295. 297. 292. 297. 11630 P 253. 260. 258. 261. 258. 261. 263. 262. 268. 266. 273. 11631 NP 287. 279. 273. 262. 273. 278. 284. 287. 294. 307. 305. 11632 P 254. 259. 260. 256. 267. 267. 267. 268. 265. 276. 276. 11633 P 301. 303. 305. 310. 311. 307. 315. 315. 328. 322. 327. 11634 P 276. 292. 293. 294. 294. 292. 299. 299. 304. 312. 318. 11.635 P 282. 295. 300. 299. 304. 300. 302. 308. 313. 313. 321. 11636 P 256. 267. 278. 277. 276. 280. 278. 287. 285. 292. 300. 11637 P 258. 268. 270. 273. 271. 276. 275. 275. 280. 285. 284. 11638 P 237. 243. 245. 247. 246. 252. 253. 255. 261. 259. 262. 11639 P 272. 277. 282. 285. 283. 285. 280. 288. 289. 297. 300. ---_-----___..--___.____________________----------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 301. 326. 309. 292. 302. 286. 328. 276. 310. 274. 301. 284. 336. 326. 321. 306. 291. 270. 302. 293 328. 313. 299. 309. 291. 335. 278. 305. 279. 314. 291. 337. 325. 356. 307. 294. 273. 309. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TAULE 27 (PAGE 22): MATEKNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11029 P 301. 300. 307. 324. 348. 361 380. 392. 11030 NP MATING NOT CONFIRMED 11031 P 324. 331. 339. 344. 357. 366. 362. 380. 410. 11032 P 315. 322. 329. 343. 367. 371. 379. 389. 409. 11033 P 304. 308. 314. 329. 335. 354. 370. 383. 394. 11034 P 312. 319. 327. 333. 349. 366. 371. 389. 401. 11035 P 298. 294. 311. 311. 324. 344. 354. 368. 366. 11036 P 342. 345. 362. 368. 385. 404. 410. 432. 446. 11037 P 282. 291. 302. 308. 329. 348. 352. 367. 374. 11038 P 313. 315. 329. 338. 352. 372. 378. 402. 408. 11630 P 276. 286. 287. 297. 304. 313. 324. 328. 327. 11631 NP 302. 305. 297. 303. 290. 292. 292. 286. 287. 290. 288. 291. 11632 P 296. 303. 304. 316. 324. 332. 344. 353. 361. 11633 P 341. 346. 356. 373. 384. 406. 420. 428. 11634 P 328. 337. 340. 353. 361. 375. 389. 401. 401. 11635 P 328. 339. 344, 353. 368. 378. 396. 412. 431. 11636 P 319. 328. 317. 340. 366. 382. 407. 415. 433. 11637 P 298. 312. 312. 324. 340. 358. 361. 381. 394. 11638 P 277. 290. 290. 304. 318. 332. 343. 352. 363. 11639 P 314. 326. 334. 342. 352. 366. 377. 396. .______.._______________________________.-.------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 300. P c-. 90 0 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 23): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS _-_..___________________________________--------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 11 12 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 271. 275. 280. 251. 269. 260. 274. 261. 260. 267. 268. 245. 277. 259. 294. 265. 257. 236. 297. 252. 266. 252. 294. 261. 297. 290. 258. NOT CONFIRMED 280. 276. 277. 282. 282. 287. 246. 244. 273. 276. 266. 271. 279. 279. 269. 269. 269. 271. 270. 283. 268. 275. 248. 246. 288. 287. 267. 266. 303. 303. 269. 271. 250. 257. 243. 248. 300. 301. 261. 264. 278. 274. 249. 252. 298. 302. 263. 267. 303. 303. 300. 300. 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. 282. 286. 295. 285. 291. 240. 243. 274. 274. 274. 276. 290. 291. 280. 279. 278. 278. 286. 291. 282. 282. 256. 256. 295. 294. 266. 272. 299. 306. 273. 279. 271. 270. 252. 263. 318. 319. 267. 275. 273. 281. 258. 263. 308. 306. 271. 273. 315. 322. 317. 312. 271. 273. _ _ -_ - - . - .- - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 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. 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. 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. 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. 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. ______ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 24): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11040 P 288. 297. 296. 312. 326. 340. 350. 361. 3 74 11041 P 313. 313. 331. 340. 358. 370. 391. 416. 11042 P 313. 310. 319. 330. 346. 359. 375. 394. 11043 NP 251. 245. 240. 245. 250. 249. 244. 233. 244. 242. 244. 243. 249 11044 P 300. 298. 308. 317. 335. 357. 368. 380. 397. 11045 P 290. 296. 312. 324. 340. 356. 370. 376. 391. 11046 P 323. 324. 336. 346. 364, 364. 388. 404. 431. 11047 P 293. 292. 310. 318. 326. 344. 356. 365. 384. 11048 P 303. 314. 321. 327. 333. 340. 354. 368. 379. 11049 P 304. 312. 319. 330. 330. 344. 353. 353. 358. 11050 P 302. 305. 317, 323. 337. 355. 375. 382. 400. 11051 P 276. 285. 292. 302. 308. 325. 345. 350. 377. 11052 P 320. 325. 340. 346. 364. 386. 400. 421. 444. 11640 P 294. 304. 302. 308. 328. 346. 360. 368. 396. 11641 P 332. 334. 345. 349. 369. 384. 393. 409. 421. 11642 P 296. 305. 314. 326. 342. 366. 382. 386. 11643 P 295. 300. 305. 313. 328. 348. 353. 373. 383 11644 P 282. 290. 297. 307. 318. 340. 355. 366. 11645 P 347 I 354. 364. 386. 398. 410. 428. 441. 450. 11646 P 290. 306. 311. 323. 341. 352. 369. 380. 384. 11647 P 310. 318. 321. 332. 347. 363. 368. 383. 11648 P 278. 294. 303. 317. 339. 342. 353. 357. 375. 11649 P 333. 334. 343. 353. 375. 391. 407. 423. 428. 11650 P 291. 297. 305. 310. 329. 342. 356. 374. 385. 11651 P 342. 352. 357. 362. 371. 390. 410. 425. 429. 11652 NP 302. 303. 307. 304. 306. 302. 300. 304. 304. 308. 310. 308. 313. . 11653 ___ P _ _ _ 293. __ _ 300. __ . _ 310. __ _ 316. .__ _ 333. __ _ 348. __ _ 367. ___ _ 3__ _7_3_ _. _ 380. __ _ _ _ _ - . - . - - - - - - - . . . - - - - - - . . . - - - - ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 25): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 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. ALL WEIGHTS WERE RECORDED IN GRAMS (G). P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION 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. 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. 3_ 1_ _0_. 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. 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. ____ 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. _-__ 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. 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. ______ PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 27 (PAGE 26): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11053 P 271. 276. 285. 299. 316. 329. 344. 366. 386. 11054 P 343. 335. 355. 356. 370. 384. 404. 425. 447. 11055 P 299. 300. 314. 325. 342. 365. 376. 388. 425. 11056 P 265. 259. 275. 280. 295. 306. 327. 336. 352. 11057 P 278. 276. 285. 298. 312. 328. 334. 345. 357. 11058 P 300. 299. 313. 328. 346. 369. 371. 389. 421. 11059 P 285. 290. 301. 310. 325. 342. 349. 370. 384. 11060 P 301. 305. 317. 327. 352. 363. 374. 392. 425. 11061 P 305. 315. 324. 336. 358. 365. 383. 394. 431. 11062 P 286. 291. 301. 313. 317. 333. 349. 366. 390. 11063 P 301. 301. 314. 324. 341. 358. 378. 389. 418. 11064 P 321. 322. 332. 337. 346. 361. 373. 385. 408. 11065 P 295. 301. 311. 321. 327. 324. 346. 359. 382. 11066 P 308. 318. 333. 342. 354. 374. 383. 408. 439. 11654 P 288. 285. 299. 307. 323. 335. 354. 375. 377. 11655 P 304. 304. 315. 328. 349. 360. 346. 371. 394. 11656 P 294. 300. 312. 326. 328. 343. 366. 370. 353. 11657 P 332. 333. 349. 360. 368. 384. 406. 418. 11658 P 292. 292. 296. 303. 312. 328. 344. 352. 362. 11659 P 286. 294. 299. 302. 316. 340. 355. 358. 362. 11660 P 296. 306. 314. 324. 342. 366. 365. 379. 11661 P 294. 305. 314. 327. 336. 350. 368. 377. 11662 P 308. 315. 321. 332. 345. 358. 366. 382. 11663 P 318. 325. 329. 343. 356. 373. 395. 409. 11664 P 274. 280. 287. 299. 312. 323. 337. 344. 11665 P 308. 314. ,325. 336. 352. 364. 380. 397. P w 11666 NP 263. 267. 270. 265. 263. 267. 266. 261. 11667 P 328. 337. 340. 350. 367. 381. 398. 416. b ALL WEIGHTS WERE RECORDED IN GRAMS (G). PP P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) w DAY = DAY OF PRESUMED GESTATION b M PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 28 (PAGE I ) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - F o GENERATION FEMALE RATS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 28 (PAGE 2) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 28 (PAGE 3): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .................................................................................................................................... DAY 1 5 RAT # DOSAGE GROUP 3 1.6 MG/KG PFOS + MEVALONIC ACID 10929 215. 284. 10936 FOUND DEAD ON DAY 2 OF STUDY 10938 314. 316. 10939 NOT PREGNANT 10941 261. SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 10942 290. 311. 11529 265. 280. 11530 330. 355. 11531 NOT PREGNANT 11532 320. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11533 292. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11534 343. 336. 11535 310. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11536 307. 320. 11537 284. 308. 11538 288. 304. 11539 255. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11540 354. 330. 11541 290. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11542 312. 298. 11543 336. 348. --.--.--.________.--____________________-..-----~~--~~~~ ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . DAY = DAY OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 28 (PAGE 4): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 ..________..._._________________________...~...~~ RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID __.___.____..__^_.______________________-.------------------------------------------.----------------------------------------------- 10943 279. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 10944 290. 282. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 10948 322. SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 10952 299. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 10955 272. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 10956 302. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11544 305. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11545 348. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11546 281. SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 11547 300. 299. 11548 NOT PREGNANT 11549 310. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11550 290. SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 11551 326. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11552 296. 310. 11553 291. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11554 272. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11555 322. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11556 SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 11557 310. SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS -----..------___-.--____________________.--~~~~---~~~~..~~ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION .. w b-- 4 4 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 28 (PAGE 5): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL,DATA - Fo GENERATION FEMALE RATS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 28 (PAGE 6): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS E'ROTOCOLI418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TRBLE 28 (PAGE 7): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL .................................................................................................................................... 10986 309. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 10994 317. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 10995 277. 282. 10996 291. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 10997 250. 261. 10998 NOT PREGNANT 11586 279. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11587 340. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11588 312. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11589 303. 290. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 11590 290. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11591 336. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11592 312. 297. 11593 272. 262. 11594 309. 299. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 11595 291. SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 11596 282. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11597 278. 275. 11598 287. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11599 278. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS ...------..-.___________________________------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION 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 28 (PAGE 8): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 28 (PAGE 9): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL,DATA - FO GENERATION FEMALE RATS DAY 1 5 RAT # DOSAGE GROUP 9 0.8 MG/KG PFOS 11009 259. 208. 11010 316. 346. 11011 325. 335. 1101.2 269. 283. 11013 277. 290. 11014 NOT PREGNANT 11015 298. 298. 11016 283. 279. 11017 NOT PREGNANT 11018 325. 324. 11610 NOT PREGNANT 11611 341. 356. 11612 288. 296. 11613 340. 364. 11614 287. 326. 11615 327. 355. 11616 296. 278. 11617 313. 326. 11618 322. 332. 11619 302. 245. -._____.._______________________________..----------------..---- ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION PRO'I'OCOI~ 418-018: 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 10): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DAY 1 5 .................................................................................................................................... RAT # DOSAGE GROUP 10 1 MG/KG PFOS 11019 307. 306. 11020 11021 11022 11023 322. 336. 287. 282. NOT PREGNANT 294. 281. 11024 11025 11026 11027 321. 285. 302. 276. 337. 314. 308. 277. 11028 310. 300. 11620 285. 275. 11621 315. 317. 11622 309. 322. 11623 308. 305. 11624 11625 307. 322. 323. 323. 11626 301. 310. 11627 284. SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11628 317. 324. 11629 325. 331. .................................................................................................................................... ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 8 (PAGE 11): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 5 _.______..__..___...____________________~~~~.~~ RAT # DOSAGE GROUP 11 1.2 MG/KG PFOS 11029 11030 11031 11032 11033 11034 11035 11036 11037 11038 11630 11631 11632 11633 11634 11635 11636 11637 11638 11639 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 c-' 00 P PROTOCOL 418 018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TAE3LE 28 (PAGE 12): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DAY 1 5 .___.-___-__..___..-____________________~.~.~~~~-~ RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS 11039 11041 11042 11043 11048 11049 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 NOT PREGNANT 297. 292. 295. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS NOT PREGNANT 277. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 296. 304. 278. 293. 304. 316. 297. 290. 294. 281. 263. 288. 332. SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 303. 316. 310. 300. 274. 279. 314. 305. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 302. 289. 343. 352. NOT PREGNANT 290. 276. ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAY = DAY OF LACTATION mvroccx,410-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 28 (PAGE 1 3 ) : MATERNAL BODY WEIGHTS - 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 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 PROTOCOI, 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 9 (PAGE 1): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - F O GENERATION FEMALE RATS .._._.._____.._...______________________~~~~~~-.-~~~~~.~~-.~ DAYS 1- 8 8 - 1 5 1 5 - 2 2 2 2 - 2 9 2 9 - 3 6 3 6 - 4 2 a 5613-63 6 3 - 7 0 7 0 - 7 7 7 7 - 8 1 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 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. 111. 119. 129. 91. 139. 92. 111. 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 29 (PAGE 2): FEED CONSUMPTION VALUES - PREMATING INDIVIDLJAL DATA - Fo GENERATION FEMALE RATS ....___.._._._.....______ 10915 151. 135. 123. 123. 10916 C 179. C 157. 10917 147. 131. 149. 127. 10918 141. 136. 130. 145. 1091 9 140. 125. 120. 136. 10920 C 138. 158. 135. 10921 152. 139. 144. 141. 10922 149. 137. 136. 135. 10923 139. 124. 145. 159. 10924 146. 131. 130. 139. 10925 142. 138. 129. 131. 10926 146. 159. 147. 145. 10927 154. 137. 134. 141. 10928 127. 124. 124. 128. 11515 116. 115. 119. 123. 11516 155. 184. 184. 158. 11517 130. 124. 119. 118. 11518 140. 162. 157. 157. 11519 137. 145. 160. 166. 11520 141. 152. 134. 132. 11521 146. 148. 146. 143. 11522 121. 136. 137. 133. 11523 126. 134. 144. 123. 11524 135. 141. 142. 135. 11525 138. 153. 168. 151. 11526 127. 135. 153. 143. 11527 130. 142. 128. 128. 11528 141. 146. 141. 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. 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. PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 2 9 (PAGE 3 ) : FEED CONSUMPTION VALUES - PREMATING INDIVIDUAL DATA - F O GENERATION FEMALE RATS DAYS 1- 8 8 - 1 5 1 5 - 2 2 2 2 - 2 9 2 9 - 3 6 RAT # DOSAGE GROUP 3 10929 122. 113. 119. 121. 117. 1.0930 131. 132. 137. 134. 129. 10931 130. 115. 110. 111. 104. 10932 138. 121. 122. 124. 128. 10933 119. 127. 121. 118. 112. 10934 149. 130. 124. 122. 117. 10935 120. 99. 105. 106. 100. 10936 FOUND DEAD ON DAY 2 OF STUDY 10937 139. 130. 116. 121. 122. 10938 143. 129. 130. 124. 135. 10939 148. 125. 118. 111. 113. 10940 132. 135. 138. C 148. 10941 133. 121. 123. 111. 115. 10942 134. 128. 131. 120. 123. 11529 120. 121. 120. 119. 112. 11530 144. 150. 153. 154. 134. 11531 154. 139. C 116. 149. 11532 130. 132. 143. 135. 146. 11533 149. 147. 136. 142. 145. 11534 146. 151. 135. 155. 155. 11535 143. 115. 130. 134. 129. 11536 141. 151. 154. 154. 147. 11537 126. 141. 138. 125. 129. 11538 136. 138. 135. 107. 119. 11539 94. 90. 119. 92. 115. 11540 129. 142. 135. 138. 140. b 1 1 5 4 1 129. 122. 115. 134. 133. P 11542 123. 223. 180. 125. 136. c-r 00 11543 162. 165. 170. 160. 156. c-r 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 5613-63 63- 70 70- 77 77- El 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. 1.15. 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. 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 d. Value was not recorded. this value. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 29 (PAGE 5): FEED CONSUMPTION VAI'UES - PREMATING - INDIVIDUAL DATA - F O GENEMTION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 42a 5623-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 5 CHOLESTEROL CONTROL __._.___..._____________________________------ 10957 122. 120. 122. 121. 135. 94. 10958 134. 140. 142. 144. 157. 129. 10959 131. 125. 129. 124. 127. 113. 10960 135. 125. 150. 146. 150. 142. 10961 114. 131. 140. 149. 163. 116. 10962 133. 138. 143. 158. 143. 127. 10963 135. 128. 133. 129. 151. 125. 10964 143. 139. 141. 136. 160. 127. 10965 124. 126. 128. 122. 122. 109. 10966 147. 142. 174. 152. 173. 137. 10967 125. 125. 129. 136. 136. 114. 10968 130. 125. 135. 136. 134. 113. 10969 129. 127. 122. 126. 127. 104. 10970 150. 148. 159. 155. 163. 124. 11558 125. 129. 126. 131. 133. 105. 11559 143. C 151. 144. C 121. 11560 134. 146. 136. 149. 152. 121. 11561 137. 145. 163. 159. 147. 115. 11562 152. 144. MORIBUND SACRIFICED ON DAY 16 OF STUDY 11563 149. 169. 166. 165. 173. 11564 128. 131. 130. 126. 134. 11565 129. 117. 117. 126. 123. 11566 130. 133. 146. 151. C 11567 153. 154. 141. 128. 176. 11568 110. 109. 141. 105. 113. 11569 133. 146. 137. 142. 155. 90PP 11570 127. 129. 126. 133. 158. P c-' 11571 143. 140. 147. 143. 161. - _ _ _ _ _ - - - _ - _ __ _ . ._- -_ . _ _ _ _ _ _ - . - _ . _ _ . _ _ _ _ _ _ - _ _ . . - . ALL WEIGHTS WERE RECORDED IN GRAMS (G). 0 c-' DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. 2 b. First value recorded after cohabitation. 0m c. Spilled feed precluded the calculation of this value. PROTOCOL, 418-018: 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 - ....- - .- - - - - - - ... RAT # .._--....___..___. 10971 120. 117. 113. 105. 10972 144. 150. 135. 135. 10973 124. 129. 127. 122. 10974 124. 141. 127. 120. 10975 130. 136. 134. 127. 10976 155. 137. 136. 121. 10977 132. 123. 117. 116. 10978 126. 122. 123. 122. 10979 125. 130. 165. 140. 10980 133. 123. 127. 113. 10981 126. 121. 121. 116. 10982 122. 123. 131. 115. 10983 153. 134. 114. 120. 10984 117. 127. 126. 127. 11572 103. 104. 105. 106. 11573 131. 129. 131. 132. 11574 127. 136. 148. 121. 11575 129. 137. 157. 133. 11576 141. 132. 164. 133. 11577 142. 129. 128. 131. 11578 140. 138. 124. 130. 11579 136. 147. 139. 136. 11580 119. 132. 126. 130. 11581 145. 133. 150. 154. 11582 134. 128. 135. 127. 11583 131. 126. 120. 127. 11584 133. 133. 133. 136. 11585 126. 142. 127. 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 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 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 29 (PAGE 7): FEED CONSUMPTION VALUES - PREMATING INDIVIDLJAL DATA - FO GENERATION FEMALE RATS 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. C 145. 134. 139. 137. 146. 136. 118. 117. 118. 122. 121. 123. 118. 118. 137. 146. 132. 126. 129. 119. 133. 118. 113. 109. 149. 125. 142. 147. 143. 129. 147, 131. 156. 161. 162. 164. 128. 131. 133. 130. 110. 115. 133. 140. 148. 141. 123. 162. 134. 140. 117. 128. 118. 118. 132. 127. 126. 125. 111. 129. 112. 112. 131. 116. 109. 113. 106. 138. 131. 140. 1.35. 134. 156. 124, 143. 98. 130. 129. 117. 126. 108. 110. 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 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. value. 2 MG/KG PFOS + CHOLESTEROL .________________________________ 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. P + 92. 00 bc-' PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 29 (PAGE 8): FEED CONSUMPTION VALUES - PREMATING INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAYS 1 - 8 8- 15 15- 22 22- 29 29- 36 RAT H DOSAGE GROUP 8 . _ . _ . _ _ _ . . _ . _ . _ . _ __ . _ _._ . _ __ __ . ._ . - _ -_ - - _ - - - _ _ - - - . - . . - . . . 10999 121. 121. 117. 122. 124. 11000 138. 135. 131. 129. 130. 11001 122. 127. 124. 116. 116. 11002 127. 131. 125. 124. 131. 11003 132. 134. 142. 150. 151. 11004 143. 148. 140. 142. 155. 11005 165. 169. 135. 135. C 11006 156. 141. C 142. 167. 11007 158. 137. 159. 165. 156. 11008 C 153. 157. 152. 153. 11600 147. 159. 139. 150. 171. 11601 148. 149. 122. 134. 148. 11602 119. 108. 122. 116. 140. 11603 138. 43. 147. 122. 136. 11604 132. 140. 143. 138. 146. 11605 150. 146. 147. 153. C 11606 138. 127. 133. 128. 130. 11607 125. 122. 130. 123. 123. 11608 126. 132. 125. 127. 135. 11609 129. 135. 150. 150. C _.______________________________________-------------.---. 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. PROTOCOLI 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6255.25) TABLE 25 (PAGE 9): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DAYS 1- 8 8- 15 15- 22 22- 29 25- 36 36- 42a 56b-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 9 0.8 MG/KG PFOS 11005 132. 112. 113. 1 1 6 . 113. 11010 C 151. 135. 158. C 11011 135. 135. 139. 141. 131. 11012 120. 123. 117. 116. 120. 11013 121. 122. 127. 124. 117. 11014 137. 133. 138. 133. 133. 11015 141. 137. 141. 140. 131. 11016 117. 112. 122. 118. 112. 11017 157. 150. C C C 11018 140. 139. 141. 153. 140. 11610 142. 135. 143. 125. 125. 11611 149. 142. 157. 146. 146. 11612 113. 122. 128. 115. 115. 11613 151. C C 160. 154. 11614 124. 138. 138. 136. 129. 11615 124. 129. 144. 144. 124. 11616 122. 132. 121. 101. 106. 11617 131. 138. 145. 148. 131. 11618 128. 131. 135. 148. 161. 11615 153. 136. 131. 107. 125. __._.___________________________________------------------ 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. w +W ul PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 5633-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 10 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. 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. PRO-IOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 5623-63 63- 70 70- 77 77- 81 RAT # DOSAGE GROUP 11 1.2 MG/KG PFOS _~.__...._.__.-..___.____________________--~.~~~~.~~~.----....--------------~~~----- 11029 140. 128. 125. 148. 133. 83. 11030 147. 137. 143. 131. 126. 108. 132. 101. 133. 64. 11031 133. 138. 143. 147. 144. 101. 11032 137. 136. 143. 141. 131. 121. 11033 133. 136. C 140. 139. 118. 11034 123. 114. 125. 124. 117. 93. 11035 130. 120. 120. 117. 124. 90. 11036 155. 142. C 124. 115. 104. 11037 121. 116. 128. 136. 141. 101. 11038 134. 132. 131. 126. 123. 103. 11630 147. 118. 128. 118. 118. 101. 11631 140. 160. C C C C 11632 125. 132. 132. 125. 119. 91. 11633 136. 146. 155. 141. 148. 118. 11634 140. 133. 142. 145. 132. 121. 11635 142. 144. 145. 152. 145. 127. 11636 128. 125. 128. 133. 130. 115. 11637 125. 126. 132. 116. 108. 94. 11638 109. 106. 119. 111. 108. 87. 11639 142. 132. 136. 125. 130. 103. _.__.___________._______________________-----------------.--.--.- 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. PROTOCOL,418-018: 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 12): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 11039 149. 143. 120. 135. 132. 11040 130. 131. 137. 137. 120. 11041 144. 134. 140. 132. 132. 11042 143. 137. 141. 130. 127. 11043 126. 126. 127. 131. 135. 11044 138. 136. 139. 125. 128. 11045 147. 131. 139. 143. 119. 11046 136. 130. C C 131. 11047 122. 116. 116. 129. 116. 11048 116. 115. 120. 125. 117. 11049 125. 127. 138. 132. 126. 11050 143. 130. 128. 134. 116. 11051 133. 122. 122. 120. 109. 11052 137. 131. 138. 135. 121. 11640 155. 127. 124. 133. 127. 11641 156. 160. 163. 146. 148. 11642 142. 138. 153. 128. 121. 11643 148. 136. 140. 123. 132. 11644 128. 126. 117. 114. 133. 11645 173. 154. 160. 146. 179. 11646 137. 131. 128. 116. 118. 11647 144. 141. 135. 130. 140. 11648 103. 98. 104. 95. 108. 11649 149. 154. 162. 147. 133. 11650 129. 129. 119. 126. 105. 11651 149. 150. 141. 133. 149. 90 11652 148. 146. 141. 142. 151. P c. 11653 158. 112. 121. 115. 116. - - - - - - - _ _ _ _ _ _ _ _ _ _ _ _ - _ _ _ _ _ _ _ _-_ - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - . . - - - 0 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 29 (PAGE 13): FEED CONSUMPTION VALUES - PREMATING - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 11053 122. 114. 126. 121. 112. 11054 142. 134. 142. 143. C 11055 122. 115. 112. 112. 111. 11056 126. 107. 101. 103. 105. 11057 120. 113. 115. 116. 109. 11058 139. 131. 139. 115. 110. 11059 138. 111. 127. 151, 122. 11060 120. 119. 124. 133. 113. 11061 132. 128. 135. 131. 127. 11062 139. 122. 128. 124. 124. 11063 127. 126. 125. 118. 114. 11064 136. 137. C 140. C 11065 103. 109. 115. 116. 106. 11066 155. 132. 127. 121. 121, 11654 127. 121. 121. 121. 111. 11655 150. 134. 116. 107. 119. 11656 C 143. 149. 126. 135. 11657 143. 139. 136. 130. 137. 11658 129. 136. 119. 122. 117. 11659 128. 116. 132. 125. 123. 11660 135. 130. 125. 114. 111. 11661 1.66. 133. 129. 118. 126. 11662 133. 135. 140. 131. 127. 11663 148. 143. 152. 141. 144. 11664 122. 126. 119. 106. 113. 11665 124. 124. 130. 113. 108. 11666 136. 124. 130. 126. 129. 11667 136. 135. 134. _ . . _ _ _ _ . - . - _ _ ..._ __ __._ . ._ _ - 132. 131. _ . _ _ ..._ 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 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 30 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10901 P 144. 157. 156. 10902 P 140. 147. 174. 10903 P 157. 162. 189. 10904 P 164. 178. 183. 10905 P 170. 171. 201. 10906 P 159. 159. 175. 10907 P 162. 169. 179. 10908 P 147. 161. 171. 10909 P 169. 147. 150. 10910 P 186. 186. 193. 10911 P 167. 153. 164. 10912 NP 173. 196. 150. a 10913 P 131. 137. 156. 10914 P 141. 139. 154. 11501 P 143. 135. 154. 11502 P 170. 169. 170. 11503 P 176. 169. 144. 11504 P 162. 176. 167. 11505 P 162. 166. 153. 11506 P 172. 165. 186. 11507 P 153. 169. 192. 11508 P 1.45. 162. 172. 11509 P 165. 174. 185. 11510 P 143. 164. 179. 11511 P 132. 137. 149. b- 11512 P 169. 185. 183. P + 11513 P 123. 131. 155. 00 11514 P 164. 163. 163. 0 0 PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 3 0 (PAGE 2 ) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS - - _ _ _ . . . . - _ __ _ _ __ _ . _ __ -_ - _ __ __ __ - _ . _ _ _ _ - . - _ _ _ . - - - . . - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 10915 P 10916 P 10917 P 10918 P 10919 P 10920 P 10921 P 10922 P 10923 P 10924 P 10925 P 10926 P 131. 147. 136. 167. 161. 144. 152. 157. 152. 163. 148. 158. 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 1 2 OF GESTATION; DEATH WAS ATTRIBUTED TO AN INTUBATION ACCIDENT 10927 P 10928 P 11515 P 11516 P 146. 151. 130. 166. 154. 160. 132. 157. 163. 185. 138. 173. 11517 P 148. 150. 152. 11518 P 156. 148. 184. 11519 P 167. 184. 180. 11520 P 150. 155. 155. 11521 P 172. 159. 166. 1 1 5 2 2 NP 139. 134. 106. 57. 11523 P 135. 138. 154. 11524 P 156. a 162. 11525 P 172. 188. 191. 11526 P 142. a 153. P 11527 P 123. 155. 176. c--L so 1 1 5 2 8 P 160. 166. 169. 0 - P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) PP DAYS = DAYS OF PRESUMED GESTATION 7 ALL WEIGHTS WERE RECORDED IN GRAMS (G). 0 a . S p i l l e d f e e d precluded the c a l c u l a t i o n of this value. W PI4 0 C-r PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 30 (PAGE 3 ) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ._._______________...-------------...-----------..------------.------------ PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 _..._________._____..-.--.----------..-...------------------------------------ RAT # DOSAGE GROUP 3 1 . 6 MG/KG PFOS t 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. 126. 119. 121. 116. 139. 105. FOUND 113. 101. 130. 167. 115. 114. 114. 139. 109. 139. 125. 133. 124. 141. 123. 119. 118. 141. 141. 127. 166, 135. 142. 131. 158. 133. 151. 124. DEAD ON DAY 138. 156. 118. 160. 139. 155. 136. 159. 120. 153. 137. 171. 146. 169. 146. 140. 122. 152. 143. 148. 168. 172. 173. 168. 191. 190. 179. 152. 2 OF 162. a 230. 161. 173. b 183. 165. 174. 174. 183. 179. 165. 144. 182. 167. 187. STUDY a 187. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value was not recorded. b. Spilled feed precluded the calculation of this value. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 30 (PAGE 4): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS 10943 P 10944 P 10945 P 10946 P 10947 P 10948 P 10949 P 10950 P 10951 P 10952 P 10953 P 10954 P 10955 P 10956 P 11544 P 11545 P 11546 P 11547 P 11548 NP 11549 P 11550 P 11551 P 11552 P 11553 P 11554 P 11555 P 11556 P 11557 P 117. 139. 165. 101. 131. 127. 165. 185. 194. a 192. 123. 139. 168. 131. 117. 176. 142. 154. 84. 135. 174. 184. 118. 155. 188. 147. 119. 138. 104. 138. 178. 127. 140. 181. 117. 153. 142. 162. 119. 142. 166. 175. 171. 189. 130. 140. 161. 147. 156. 164. 156. 116. 97. 61 134. 140. 164. 119. 132. 156. 140. 149. 183. 136. 146. 167. 133. 146. 149. 131. 137. 142. 145. a 215. 144. 143. 168. 136. 151. 138. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WELGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 3 0 (PAGE 5 ) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 . 7 - 14 14 - 2 1 2 1 - 25 RAT # DOSAGE GROUP 5 CHOLESTEROL CONTROL ...-...--.-.____________________________~..~~~~ 10957 P 10958 P 10959 P 10960 P 10961 P 10962 P 10963 P 10964 P 10965 P 10966 P 10967 P 10968 P 10969 P 10970 P 11558 P 11559 P 1 1 5 6 0 NP 11561 P 11562 11563 P 11564 P 11565 P 1 1 5 6 6 NP 11567 P 11568 P 11569 P 11570 P 11571 P 119. 152. 160. 185. 174. 153. 183. 161. 144. a 170. 160. 152. 164. 126. 191. 140. 191. MORIBUND a 159. 155. 178. 176. 135. 161. 132. 186. 153. 168. 159. 184. 165. 198. 210. 215. 171. 191. 153. 172. 178. 207. 183. 203. 142. 167. a 174. 165. 153. 166. 184. 193. 178. 142. 161. 196. 176. 127. 149. 177. 162. SACRIFICED ON 203. 192. 156. 156. 156. 168. 153. 114. 182. 207. 148. 152. 167. 167. 146. 146. 183. 203. DAY 69. 16 OF 74 STUDY P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . a. Spilled feed precluded the calculation of this value. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 6 ) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS .___________....________________________~~~~~~...~.- PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL - - - - - . . . . - _ __ __ _ _ _ _ _ _ __ .__ __ _ . _ __ _ . _ . - . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ~ 10971 P 127. 122. 149. 10972 P 151. 170. 198. 10973 P 122. 147. 160. 10974 P 137. 157. 186. 10975 P 132. 133. 174. 10976 P 120. 128. 158. 10977 P 117. 128. 139. 10978 P 125. 134. 181. 10979 P 155. 170. 179. 10980 P 118. 130. 164. 10981 P 94. 139. 166. 10982 P 120. 138. 199. 10983 P 133. 146. 10984 P 147. 168. 11572 P 116. 139. 151. 11573 P 146. 156. 175. 11574 P 126. 137. 176. 11575 P 141. 168. 181. 11576 P 127. 166. 175. 11577 P 146. 158. 180, 11578 P 147. 150. 178. 11579 P 149. 153. 160. 11580 P 137. 171. 191. 11581 P 163. 144. 188. 11582 P 146. 151. 172. 11583 P 125. 156. 173. 11584 P 135. a 160. 11585 P 113. 141. 164. . . _ . _ _ _ _ _ _._ . _ _-.--_ . - __ __ __ __ . ...._ ._._ _._ _ _ __ _ . ._ _ . .._ - . . -_ - - _ -.-_ _ . _ _ _ _ _ - . . - . . - - - - - ~ ~ ~ - - . - .- --- - - - - - - . - - - - - - - P = PREGNANT NP = NOT PREGNANT (VALUES EXCL9UDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION AL,L WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'Ss w m NUMBER: T-6295.25) TABLE 30 (PAGE 7): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 .................................................................................................................................... RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL __-_.___.__...._._______________________.~~~~~~~~~--..-- 10985 P 134. 156. 182. 10986 P 114. 136. 188. 10987 P 131. 140. 197. 10988 P 118. 138. 154. 10989 P 116. 129. 161. 10990 P 124. 146. 159. 10991 P 117. 136. 154. 10992 P 110. 135. 168. 10993 P 116. 161. 188. 10994 P 120. 135. 202. 10995 P 127. 152. 181. 10996 P 122. 131. 167. 10997 P 112. 129. 155. 10998 NP 135. 123. 127. 86 11586 P 131. 152. 158. 11587 P 146. 169. 195. 11588 P 118. 141. 11589 P 156. a 202. 11590 P 142. 158. 158. 11591 P 143. 172. 194. 11592 P 137. 138. 179. 11593 P 118. 139. 154. 11594 P 125. 150. 185. 11595 P 148. 172. 184. 11596 P 154. 150. 177. 11597 P 138. 155. 177. 11598 P 133. 138. 165. 11599 P 108. 139. 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. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 3 0 (PAGE 8 ) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS -_____..._.__...________________________-.~~~~~~~.~~~ PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 .................................................................................................................................... RAT # DOSAGE GROUP 8 0.4 MG/KG PFOS 10999 P 142. 170. 141. 11000 P 160. 168. 174. 11001 P 132. 147. 174. 11002 P 165. 163. 188. 11003 P 160. 155. 212. 11004 P 159. 180. 166. 11005 P 167. 165. 140. 11006 P 158. 177. 171. 11007 P 205. a 199. 11008 P 169. 153. 179. 1.1600 P 176. 170. 169. 11601 P 169. 169. 173. 11602 P 148. 148. 164. 11603 P 159. 161. 183. 11604 P 144. 151. 172. 11605 P 164. 169. 150. 11606 P 149. 153. 168. 11607 P 133. 147. 162. 11608 P 166. 155. 161. 11609 P 161. 180. 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. PROTOCOL 418-01a: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 30 (PAGE 9): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 66. 69. 90 0 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 30 (PAGE 1 0 ) : 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 10 1 MG/KG PFOS ..--___.-.--___...---~~.~..~~-~~~~~~~.~~~.~~~~~..~~~~ 11019 P 136. 122. 171. 11020 P 147. 152. 173. 11021 P 162. 163. 195. 11022 NP 155. 132. 107. 72. 11023 P 140. 153. 179. 11024 P 169. 183. 207. 11025 P 151. 143. 170. 11026 P 140. 161. 166. 11027 P 124. 162. 177. 11028 P 133. 144. 190. 11620 P 140. 95. 169. 11621 P 170. 147. 170. 11622 P 159. 169. 186. 11623 P 135. 152. 167. 11624 P 150. 168. 188. 11625 P 164. 167. 185. 11.626 P 129. 147. 160. 11627 P 145. 156. 119. 11628 P 127. 159. 182. 11629 P 136. 150. 173. _________.__________..------.--.--------------.-------------------.----- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY 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 - 14 14 - 21 21 - 25 RAT # DOSAGE GROUP 11 1.2 MG/KG PFOS _______._______._.______________________~~~~~~~~..~ 11029 P 135. 185. 204. 11030 NP MATING NOT CONFIRMED 11031 P 144. 157. 169. 11032 P 137. 164. 195. 11033 P 134. 174. 197. 11034 P 124. 134. 162. 11035 P 128. 149. 166. 11036 P 140. 151. 177. 11037 P 117. 146. 166. 11038 P 139. 144. 177. 11630 P 137. 142. 147. 11631 NP 153. a 8 .b 11632 P 114. 141. 150. 11633 P 161. 165. 195. 11634 P 153. 164. 153. 11635 P 161. 167. 187. 11636 P 151. 169. 193. 11637 P 128. 134. 147. 11638 P 136. 140. 164. 11639 P 128. 154. 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. b. Value appeared incorrectly recorded and w a s excluded from group averages and statistical analyses. P _.L c. Value appeared incorrectly recorded and cou1.d not be calculated. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 30 (PAGE 12): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS _____._____.________----.----------.------.------.-------.--------------.--- PREGNANCY STATUS DAYS 0 - 7 7 - 14 14 - 21 21 - 25 .................................................................................................................................... RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS ._-____.._______________________________-.------.-----..---------- 11039 NP MATING NOT CONFIRMED 11040 P 103. 126. 188. 11041 P 137. 152. 11042 P 122. 149. 11043 NP 95. 106. 103. 73. 11044 P 119. 147. 195. 11045 P 139. 152. 180. 11046 P 129. 152. 173. 11047 P 124. 129. 181. 11048 P 133. 160. 152. 11049 P 141. 150. 186. 11050 P 121. 134. 192. 11051 P 117. 142. 154. 11052 P 141. 153. 202. 11640 P 128. 143. 169. 11641 P 153. 168. 166. 11642 P 122. 131. 167. 11643 P 131. 145. 173. 11644 P 124. 136. 165. 11645 P 171. 208, 211. 11646 P 141. 151. 186. 11647 P 141. 176. 182. 11648 P 113. 126. 150. 11649 P 152. 158. 182. 11650 P 133. 120. 171. 11651 P 152. 175. 187. 11652 NP 181. 132. 116. 73. 11653 P 133. 137. 157. --- - - - - - . - - - - . . - - _ . _ . - . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ . ~ ~ . . - ~ ~ . . . ~ . . ~ ~ ~ ~ . ~ - P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOI, 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 30 (PAGE 13): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS 11053 P 106. 128. 172. 11054 P 127. 154. 191. 11055 P 130. 152. 195. 11056 P 114. 125. 154. 11057 P 127. 139. 186. 11058 P 123. 130. 189. 11059 P 115. 135. 155. 11060 P 115. 135. 195. 11061 P 133. 162. 198. 11062 P 125. 149. a 11063 P 122. 125. 177. 11064 P 171. 271. 134. 11065 P 114. 145. 159. 11066 P 134. 170. 206. 11654 P 128. 144. 166. 11655 P 158. 145. 171. 11656 P b 153. 159. 11657 P 140. 156. 180. 11658 P 131. 137. 139. 11659 P 120. 123. 141. 11660 P 119. 133. 175. 11661. P 133. 146. 161. 11662 P 148. 140. 164. 11663 P 153. 163. 184. 11664 P 126. 124. 147. 11665 P 123. 150. 171. 11666 NP 1.49. 106. 99. 63. 11667 P 149. 146. 166. .._______..________.____________________----------.--~---------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value was not recorded. b . Spilled feed precluded the calculation of this value. PROTOCOL 418-018: 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 P c-' 9" 0 w PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 3 1 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS w tL + P PROTOCOL,418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 31 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - F O GENERATION FEMALE RATS _ _ _ _ _ _ _ . _ __ . ._ __ _ _ _ . _ _ . _ __ _ _ . _ _ _ . _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - . - - - - - - - DAY 1 - 5 ______.._____._.________________________--..------------------.- RAT # DOSAGE GROUP 3 1 . 6 MG/KG PFOS + MEVALONIC ACID __._.__________...______________________.~~~~~~-~~.~~ 10929 79. 10936 FOUND DEAD ON DAY 2 OF STUDY 10938 58. 10939 NOT PREGNANT 10941 SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 10942 107. 11529 88. 11530 90. 11531 NOT PREGNANT 11532 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11533 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11534 65. 11535 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11536 a 11537 104. 11538 76. 11539 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11540 49. 11541 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11542 56. 11543 115. _________-._____________________________---------..-------.---- ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . DAYS = DAYS OF LACTATION a. Spilled feed precluded the calculation of this value. PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 31 (PAGE 4): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DAY I - 5 .................................................................................................................................... RAT # DOSAGE GROUP 4 2 MG/KG PFOS + MEVALONIC ACID 10943 10944 10948 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 10952 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 10955 10956 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11544 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11545 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11546 SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 11547 55. 11548 NOT PREGNANT 11549 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11550 11551 SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11552 65. 11553 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11554 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11555 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11556 SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 11557 SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS .-.-..----.--___________________________.-.-.------------------------ ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . DAYS = DAYS OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 3 1 (PAGE 5): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 31 (PAGE 6): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS ___________..___________________________...-.-.---.---....-.----- DAY 1 5 - .____.-.-_______________________________..--.-..-.-...--.---.------ RAT # DOSAGE GROUP 6 1.6 MG/KG PFOS + CHOLESTEROL __._.__..____.._________________________-.----.-------------------- 10971 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 10973 SACRIFICED ON DAY 4 OF LACTATION DUE TO NO SURVIVING PUPS 10977 66. 10979 105. 10983 99. 10984 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11572 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11573 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11574 57. 11575 119. 11576 87. 11577 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11578 89. 11579 51. 11580 70. 11581 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11582 58. 11583 83. 11584 109. 11585 42. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS _-__________._.___._____________________-~~~~~..~~~ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 31 (PAGE 7): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS .................................................................................................................................... DAY 1 - 5 RAT # DOSAGE GROUP 7 2 MG/KG PFOS + CHOLESTEROL 10986 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 10994 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 10995 56. 10996 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 10997 a3. 10998 NOT PREGNANT 11586 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11587 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11588 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11589 53. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 11590 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11591 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS 11592 67. 11593 39. 11594 50. SACRIFICED ON DAY 5 OF LACTATION DUE TO NO SURVIVING PUPS 11595 SACRIFICED ON DAY 1 OF LACTATION DUE TO NO SURVIVING PUPS 11596 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11597 61. 11598 SACRIFICED ON DAY 3 OF LACTATION DUE TO NO SURVIVING PUPS 11599 SACRIFICED ON DAY 2 OF LACTATION DUE TO NO SURVIVING PUPS __--.-_____---___..-____________________--.------------------------ ALL WEIGHTS WERE RECORDED IN GRAMS (G). DAYS = DAYS OF LACTATION PROTOCOL,418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) : MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS RAT # DOSAGE GROUP 8 0.4 MG/KG PFOS .................................................................................................................................... 10999 11000 11001 11002 11003 11004 11005 11006 11007 11008 11600 11601 11602 11603 11604 11605 11606 11607 11608 11609 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 PROTOCOL 4 1 8 - 0 1 8 : ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T - 6 2 9 5 . 2 5 ) TABLE 3 1 (PAGE 9): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 31 (PAGE 10): MATERNAL FEED CONSUMPTION VALIJES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DAY 1 - 5 .______..._____..._.____________________~~~ RAT # DOSAGE GROUP 10 1 MG/UG 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 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'S STUDY NUMBER: T-6295.25) TABLE 31 (PAGE 11): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS (SPONSOR'SSTUDY NUMBER: T-6295.25) TABLE 31 (PAGE 12): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS DAY 1 - 5 _.____._._..__._________________________------....-----..------------------ RAT # DOSAGE GROUP 12 1.6 MG/KG PFOS .._...-__.._--______.--------------...----.-.-.----------------------------------- 11039 11041 11042 11043 11048 11049 11640 11641 11642 11643 11644 11645 11646 11647 11648 11649 11650 11651 11652 11653 NOT PREGNANT 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 418-018:PAGE B-225 EE Egil Ei gg PlEi gE P g Elfiiom jefoE e : : i :i so m EREEuR iERE Zi 27 i i%ies sez 2mees gg; 2! glimmer pi iol tRR eRe RRRRR BRE 2! 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CoH BronNEosiI lelsln gg iif % tim in Dg o LarEdRsRwap niinREE ginff HaE dn de i a C 2 FRE l HE iHEm EEE b E tiie IEEE EE i Hoo iI deii, iiH Hilt 5 HE g 2 ig8i'% z3 418-018:PAGE B-311 !Epo oCo,BdgF Ed dlil itfaf TPoELEEnEdEmim dEnR sSpulInl EaaeEedihi dn if ) 0 [HEoRo eg FA ge Fide Pog iahiaE :=f i i = Go i iiH: 418-018:PAGE B-312 Dg LEER I g Lo o5g 48 ELE dig iz 28 IF [pid a Po pl aE aga ih Cb REEDHLGE HE BI ih i g Hzog = 5Hi EPRike izn 418-018:PAGE B-313 Don Hig oel iBoiEE i BE iORE i gait REg EEiLns id a iE Huigiay 5DEo o HE parddas aaFodgg lggf uslliid flinis]g, i # iE i gg. 3 Bx 8 ied g Hofp iz #igi Pr 5 ; in is 418-018:PAGE B-314 eo pb REEL po Bgedads Bifpimeidti og|g Doi EE EEE eee ele i i Igo. a delsdg ese fa e dns 20i s fit dsn gn i. IESE RpER fA pad HE DflEEE i gd IglE asng BtEoonEf pTaHtIgiriliidEy By i33 ihr | gd edi isd 3 3 zg; i$ 418-018:PAGE B-315 : y id $138 Po LHL HRE EEE Pub eh Maa eo EMER s i Bg BSR Ff EE : i EH alria i 53% Bore ER: Fg fF 5 EE i f bob ogi oul nu hoe gEf gd of HH i. igh inf oBEED oad dE mam Eitan wl dd dd i g BB EuEf gsNgs fBdEaE; Bdcef:oRBE @PRdg gRi dfs ihe i a H Se ECE 7 - oo - - - TIRE Si. IE BFRLE I giiet Pik = p2o Hi Pigg gE dis 0 EE cig I fig SIPo osEiEaHHiakens faBiiggrgr gmiid; g 1 iy% I Pe F i`41 Bi Feri H HEi N T.ERE. kh FiEtRe (i58 [HI i FM 1s g REE : 418-018:PAGE B-316 APPENDICX PROTOCOL AND AMENDMENTS PRIMEDICA 418-018:PAGE C-1 F L T hT a a.s PROTOCOL 418-018 `SPONSOR'S STUDY NUMBER: T-6295.25 STUDY TITLE: PURPOSE. One Generation Reproduction Study of PFOS - Mevalonic Acid/Cholesterol Challenge and NOEL Investigation in Rats CThoeaptumraponsaetsoofnthoifsmsetvuadyloanrie taocdteotrerhmeilneeswehreetlhecranor not prevent or antagonize the adverse maternal and fetal effects orn. (reduceddmeactreeransaeldbsodiynwweeiigghhttgaainnd, dienccrreeaasseeddppeurpcesnutrvival 2o7b5s.e0r0v6e.d3in aSphrdeyvinouusmbsteurdyT6(2Ar9g5us9)Reasnedar1chbePtreortodceofline the no observed effect level (NOEL). TTEESSTTINHGOEFAACCILLITTYE: STUDY DIRECTOR: SPONSOR: AGFr0og3ruaSsnnaRemee,snePyaarDmcinvheey.LiaabBonurtalstkori1in8eA0gs,44I-nc1.267 Telephone: (215) 443-8710 Telefax: (215) 443-8587 RReaeyonmaontnadyoGD.irrYeGocprtkio,moPefhd.RDEa.a,saDcrAocBmnT 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 418-018:PAGE C-2 REGULATORY CITATIONS: Protocol 41P6a.g0e128 Study Design as Modification of: U.S. Food and Drug Administration (1994). Intemational Conference on Harmonisation: Guideline on detection of toxicity to reproduction for medicinal products. Federal Register. September 22. 1994, Vol. 59, No. 183. U2.1S.CFFoRodPaarntd58D.rug Administration. Good Laboratory Practice Regulations: Final Rule. Japanese Ministry of Health and Wellare (1997). Good Laboratory Practice Standard for Satety Studies on Drugs, MHW Ordinance Number 21. March 26, 1997. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315: 28 October): 1-17. REGULATORY COMPLIANCE: This study wil 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 Facilty's Quality Assurance Unit (QAU) will audit the protocol, the raw data a`atntdhtehTeersetpionrgt,FaacnidlitwyilliniancspceocrtdcarnitciecawlipthhatsheesStofantdhaorsde pOopretriaotnisngofPtrhoecsetduudryescoonfdAurcgtuesd Research Laboratories, Inc. Should any portion of the study be conducted by a subcontractor or by the Sponsor, the QanAdUrefoprortthsatfofractihlatty will conduct critical phase study portion according to inspections and audit respective results the SOPS of that facilty. Such critical Dpihraecsteori.nsTpehcetidoantersepoofrttsheanidnsrpeepcotritonasudaintds wriellpobret ssuubbmmiitstseidonbsywtihllatbfeaciinlctoyrtpoortahteeSdtiundtyo a QAU Statement generated inclusion in the final report. by that facility and provided to the Testing Facilty for The final report wil include a the report accurately reflects compliance statement the raw data obtained signed by during the the Study Director that performance of the study aShnodutlhdatsiagllniafpipclaintcadbelveiaGtLioPnsrefgruolmatGioLnPs wreegruelaftoilolnosweodcciunr,theeaccohndwiulcltbeofdtehsecrsitubdeyd. in detail, together with how the deviation might affect the quality or integrity of the study. 413-018:PAGE C-3 STUDY SCHEDULE Protocol 4P18a.g0e1s8 See ATTACHMENT 1 to the protocol. | TEST ARTICLE AND VEHICLE: Identification: Test Article: Name: Physical Description: LovBatch Number: Specific Gravity: Pury: Expiration Date: Perfluorooctane Sulfonic Light-colored powder. Acid Potassium Salt (PFOS). 217 06 N98A.9% Information on the with the Sponsor. identity, composition, strength and purity of the test article is on file Control Atticles: Name: Physical Description: Lot Number: PuSrpietciyf:ic Gravity Expiration Date: Mevalonic White with Acid. dark yellow cast, semisolid. 0N7A0K2603 Approximately August 2004 97% Name: Physical Description: Lot Number: Specific Gravity: Purity: Expiration Date: Cholesterol. White powder N1A19HO218 95% July 2004 mInafionrtmaaitnieodn ionnthteheriadwendtaittya,.composition. strength and purity of the control articles will be Vehicles 0(R.O5D%ITHw;e0)e. nSu8p0pliinerreavnerdsleotosidmeonstiifsicmateimonbrofaTnweepernoce8s0setdo deionized water be documented in the raw data. Reverse Osmosis Membrane Processed Deionized Water (RODI H;0) 413-018:PAGE C-4 Protocol 4P18a.g01e8 1N0eibteheprretsheenStpionntshoervneohrictlhees SthtautdywoDuilrdecitnotreirsfearweawriethotfhaenryespuolttesntoifalthciosnsttuadmyi.nants likely Therefore, no analyses other than those mentioned in this protocol will be conducted. `Safety Precautions: Gcolaotveasr,edtuosbte-mwisoUmHEPdAu-rfinigltfeorremdulmaatsiokn, parpeppraorpartiiaotneaenyde dproostaegcetioandmainnidstarautniiofno.rm/Tlhabe. Material Safety Data Sheet (MSDS) is attached to the protocol (ATTACHMENT 2) Storage: Bulk Test Article: VCoenhtircolle ACrotmicploenCenotmsp:onents PPrreeppaarreedd TVeehsitcAlreti(c0le.5% Tween 80): Formulations: Room temperature. Room temperature. Room temperature. Room temperature. Frozen (:20C) Al test article shipmentsto the Testing Facility should be addressed to the attention of tJuellieapnhGounlebinnusmkbi,erM.anager of Formulations, at the previously cited address and `Shipments should cartons should be include labeled iapnpfroorpmraitaitoenlyc.onTcehrenirnecgipsiteonrtasgheocuolnddibteionnostiafinedd shipping in advance of shipment. FORMULATION: Frequency of Preparation: Formulations of PFOS (suspensions) will be prepared weekly at the Testing Facil. The vehicle (0.5% Tween 80) wil be prepared weekly. Mevalonic acid will deionized water. be prepared twice daily in reverse osmosis membrane processed Cholesterol wil 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. 418-018:PAGE C-5 Adjustment for Purity: Protocal 41P8a.g0e1s8 The test and calculations. control articles will be considered 100% pure for the purpose of dosage Testing Facility Reserve Samples: `The Sponsor the course of will this reserve study. a sample (1 The Testing g) of each Facilty wil lot of the bulk test reserve a sample article (1 g or used during 5 mL) of setaucdhy.lotSaofmpthleescownitlrloblearsttioclreedanudndveerhitchleepcroemvipoounselnytcsituedsecdondduirtiinongst.he course of this ANALYSES `thSeamcpoluersseadodfittihoenasltutdoy.those described below may be taken if deemed necessary during Bulk Test Article Sampling: NInofoarnmaaltyisonesoonf tthhee sbtualbiklitteystofartthieclbeuwlikll tbeset caortnidcuiectisedondufrilienwgitthhetcheouSrpsoensoofrt.hisTshteudy. Certificate report of Analysis is on file with the Sponsor and a copy will be included in the final Analyses of Prepared Formulations: Stabilty: Sctoanbcielinttyradtaitoansfoar npdrecpoanrdeidtitoensst article of this formulations study are on bracketing the range file with the Sponsor of and will not be wdeeetkelrymiantetdhdeuTreisntgitnhgeFcacoinldituyc.t of this study. Test article suspensions will be prepared Homogeneity Analyses: Homogeneity course of this of the study. test article A syfinge in prepared will be used suspensions to withdraw will be verified samples (5 mL during each) the from the tEoapc,hmisdadmlpelaen(d5 bmoLt)towmilofbethdeivhiidgehdesitntcootnwcoenatlriaqtuiootns oanndthpelaficrestddianytoogflparsespacroanttiaoinn.ers, aolniequooft 2(3mmLL)anwdillobnee roeft3aimnLed. atOntheeaTleisqtuoitng(2FamcLi)ltwyilalsbae bsahcipkpuepdsfaomrplaena.lysBiasc;ktuhpe other samples will be Testing Facility stored under the upon the request previously cited of the Sponsor. conditions and discarded at the 418-018:PAGE C-6 Concentration Analyses: Protocol 41P6a.g0e1s8 `ofCotnhciesnsttruadtyi.onAofsytrhienpgerewpilalrbede tuessetdarttoicwlietshudsrpaewnssiaomnpslewsill(5bemLvereiafciehd)dfurroimngeathcehcourse concentration gestation and once during each of the postnatal. Each sample following periods (5 mL each) will of be the study: premating, divided into two aliquots and splhaicpepdedinftoorgalnaaslyssciosn;ttahieneortsh,eronaelioqfuo2t m(3LmaLn)dwiollnebeofre3tmaiLn.edOantethaeliTqeusotti(n2g mFLac)ilwtiyl absea backup sample. and discarded at BthaeckTuesptsinagmpFlaceisliwtiyllupboensttohreerdeuqnudesetr otfhethpereSvpioonussolry.cited conditions. `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 willbe notified in advance of sample shipment. DISPOSITION aPrrteicplaerweidlfboermrueltautrinoendstwoillthbeeSdtiusdcyarMdoenditaotrtahtetTheestpirnegviFoaucsillytyc.iteAdllardedmraeisnsi.ng bulk test TEST SYSTEM: Species/Strain and Reason for Selection: T1)hethiCsrs:tCraDi@n(oSfDr)atIhGaSs BbReeVnAdFe/mPolnusstrraattewdatso sbeelescetnesditaivsetthoerTeepsrtodSuycstitveemabnedcause: daenvdedleovpemleonptmaelnttoaxlintsoxaincidtyheavsalbueaetnionwsi:de2l)yhuissteodritcahlroduagthaouatndinedxupsetrryiefnorcereepxirsotduactttihvee Tstersatiin;nganFdac4i)litthyis';st3r)aitnhehatsestbeaertniculeseisd pihnaprrmeavciooluosgirceaplrloyduaccttiivoen isntutdhieesspoefciPeFsOSa.nd 418-018:PAGE C-7 Number: Protocol 4P1a8g0e178 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 110 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 liters. 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 andor truck from Charles River Laboratories, Inc. to the Testing Facilty. 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 Facilty'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. 1 Generation: Pups vill not be individually identified during lactation: all parameters will be evaluated in terms of the litter. 418-018:PAGE C-8 ANIMAL HUSBANDRY: Protocol 41P6a.g0e1s8 Al cage and Use sizes and housing conditions of Laboratory Animals". are in compliance with the Guide for the Care Housing: Eo Generation Rats/F1 Generation Litters: eFxocgeepntedruartiinogn trhaetscowihlalbbietatinidoinviadnuadllpyoshtopuasretduminpsetraiiondlse.ssDsutreienlgwciorhea-bbiottattioomne,d ecaacghespair of rats will presumed be housed in gestation, Fo the male rat's cage. generation female ratBsegaisnsniignngednotolnataetrurtahlandedliavyer2y0wiolfl be individually housed in common nesting box nesting boxes. Each during the postpartum dam and period. delivered liter will be housed in a Nesting Material Bedding delivery. material (bed-0'cobs?) will be supplied to female rats assigned to natural BAneadldyisnegswiflolrbpeoscshibalnegecodnatsamoifntaetnioans anreececsosnadruycttoedkeseepmit-haennaunailmlaylsanddrydaoncdumcelenatne.d in the raw data Room Air, Temperature and Humidity; f`Trheeshaaniirmtahlatrhoaosm biseeinndpepaesnsdeednttlhyrosuugphpl9i9ed.9w7it%hHatEPleAasftittteerns.chRaonogemstpeemrpehroautruorfe 1wi0ll0b%e maintained will also be maton6i4toFre(d18coCns)ttaont7l9yaFnd(2m6aiCn)taanindemdoantit3o0r%edtoco7ns0t%antly. Room humidity Light: An automatically controlled 12-hour ight: maintained. Each dark period will begin 12-hour at 1900 dark fluorescent hours EST. light cycle will be Diet: RaadtlsibwiitlulmbfergoimveinndCievritdiufailefdeeRdoedres.nt Diet #5002 (PMI Nutrition Intemational) available 418-018:PAGE C-9 Protocol 41P8a.g0e189 Water: Water wil be available adlibitum from individual bottles attached to the cages or from an automatic watering access system. All water wil be from a local source and passed through a reverse osmosis membrane before use. Chlorine will be added to the processed water as a bacteriostat; processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. Contaminants NtoeibteheprretsheenStpionntshoerdnroinrktihneg wSattuedry oDrirtehcetonresistianwgamraeteorfiaalnyatpolteevnetlisalthcatonwtoaumlidnainnttesrfleirkeely awtitehxtthreemreelsyulltsowofletvheilssstinudtyh.eTcheretiSfipeodndsieot.r isThaiwsarloew-olfevaelpoctoennttiaamlicnoantitoanmiwnilalntnoptresent 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 willbe. assigned to individual housing on the basis of computer-generated random units. After accimation, 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 cohabitation, dosage group, one male rat pceornfseemcautlievreat.ordTehrewiclolhbaebiutsateidontopaesrsioidgnwirllatcsotnosist 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 altemate male rats that have mated and will remain in cohabitation for a maximum of seven addtional days. The first eight female rats per dosage group a confirmed date of mating will be assigned t(ogCraoeuspasr1e1a0n-7s, ec1t2iaonnidng13o,nRedpalyic2a1teof1) with presumed gestation. The remaining female rats (including those with no confirmed mating date) will be permitted to naturally deliver ters. 418-018:PAGE C-10 Protocol 4Pa1g8e01180 E1 Generation Pups: 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) itis 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: (Femmaaxliemuramtsofwi1ll4bdeaygsi)veanndthceotnetsitnauritnigcltehdraoiulgyhbedgaiynn2i0ngof4p2rdeasyusmebdefgoersetactoihoanbi(traattison 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 testicontrol article; such animals may not receive any or all of their daily dosages) on the day of parturition. 1 Generation: F1 generation pups wil not be directly given the test article, but will be possibly exposed 0 the test article during matemal gestation (in utero exposure) or via matemal 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. soispace cl FS Dosage Levels. Concentrations and Volumes: ome Tre eer ] [r[eoom| eavnoisr[|0000T|ooonmee|roevsorteonr| e|omesrmn]| HERDER |[mem]=[ mem | vemos| me | Tamm |=[men| ore |e, | |[om [=[ame|owe | | EEE E E a H Tome|E e[0[R omenT E =] [CoTeoersmreess[| 00T T oTorr rT es| | u ] ] CCoo TTiomworra[ TooTToTTearmaere||w]] [CoTomoresToTT orp w] EE a aE l ET [| mLcomomoemmo | ]To0 aT | |aT+T Te rem oem eea]]| CCCooomeT T | e oTTTeoe aeT T Tr t rTTTeeenmmae]]] CCorem r TTeT Ta rT Tesmee]] CoeTeTea TtTee] TET II 418:018:PAGE C-12 Protocol 4P1a8g.e01182 TESTS, ANALYSES AND MEASUREMENTS - Fo GENERATION: Viability: All Periods: Atleast twice daily. Clinical Observations and/or General Appearance: Acclimation Period: Atleast once. Dosage Period: Prior to dosage administration, approximately one hwoorukrianfgtedratyhe(aspepcroonxidmdatoeslaygeonaendhoautrtahfeteerntdheofthtihred dosage). Day of Sacrifice: Once. Matemal Behavior: Days 1 and 5 postpartum. Any observed abnormal behavior will be recorded daily. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or Study Monitor. Body Weights: Acclimation Period: Atleast once. Dosage Period: Weekly to cohabitation. Daily during presumed gneastutraatliodnelainvedryo)n Day 1 postpartum (rats assigned to Sacrifice: Terminal weight. Feed Consumption Values (recorded and tabulated) Dosage Period: Weekly to cohabitation. On days 0, 7, 14, 21 and 25 (if necessary) of presumed gestation and on days 1 and 5 postpartum (rats assigned to natural delivery). Feed consumption values may be recorded more frequently than cited above if it is cnaegceeswsiatrhyotnoerefpeleednijasrh, trheeplfeenedi.shmDeunrtinogf ctohheabfieteadtijoanr,s wwihlebnetdwoocuramtesnotcecdu.pyIntdhievisdauamle values will not be recorded or tabulated. 418-018:PAGE C-13 Protocol 4P1a8g.e01183 Mating Performance: 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: Fentiity Index (percentage of matings that result in pregnancies). Gestation Index (percentage of pregnancies that result in birth of live litters). Number of offspring per liter (ive and dead pups) Number of implantation sites. General condition of dam and liter during the postpartum period. Viabilty Indices (percentage of pups bom that survive 5 days). Caesarean-Sectioning Observations: Eight rats from dosage groups 110 7. 12 and 13 (Replicate 1) will be Caesarean sectioned on day 21 of presumed gestation. The fetuses will be removed from the uterus and pooled byliter for body weights. The rats will be examined for number and distribution of Corpora Lutea. Implantation Sites. [Placentae that appear abnormal (size, color or shape) will be noted in the raw data), Live and Dead Fetuses. (aAtleivrem ffeettuuss itshadtefdioneesd naostorneesptohantdrteossptoinmudlsi taondstitmhualit:isandoteamdarfkeetudslyisaduetfoilnyezdeda;s 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 tis grossly evident that organogenesis has occurred: if this is not the case, the conceptus is defined as an early resorption.) 418-018:PAGE C-14 Protocol 4Pa1g8e01184 Fetal Observations: Body Weights: Fetuses wil be pooled by litter and litter body weights will be recorded. (Only body weights of live fetuses wil be recorded.) Blood and Tissue Samples (See ATTACHMENT 4for 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. copies of these weights will bTeheisneclwuediegdhwtisthwiltlhebepadcokciunmgeinstt epdrioinr ttoheshriapwmednatta., aSnadmple tubes will be labeled with the study number. litte 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 liter will be placed into EDTA-coated tsuabmepsleasndwiltlheberesmpauinniinngafecteanltrbilfouogdewailnldbtehetrraenssufletrirnegdsienrtoumse(rduivmidseedpairntaototrwotubes. The. 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 iter will be divided into three samples of equal number until shipment (when possible). for analysis. The Two of the remaining samples will be frozen and sample will be flash frozen stored (<-20C) 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. 418-018:PAGE C-15 Protocol 4P1a6g.e01185 The remaining serum and liver samples will be analyzed for LDL, HDL, total cholesterol and triglycerides. These samples will be shipped to: Or. 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 WToerlceepshtoenre,: Ma(s5s0a8c)h8u9s0e-t0t1s5101608 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 Rdeactaspiwtilaltiboen.sacWrhifeincedpobsysicblaer,bornatsd.iopxuipdes aasnpdhyfxeiattuisoens.wilFlebteusseascrwiiflicbede asancdritfiiscseudesvia collected at approximately the same time each day. 418-018:PAGE C-16 NECROPSY - Fo GENERATION: Protocol 4P1a8g.e01186 Gevraolsusatlieosnio(nastwaibllleboefrreatnaidnoemd uinnintesutwirlallbbeufufseerdedto1s0e%lefcotromanleincofnotrroplosgsriobulep fruattuarsesigned toordCearetsoarperaonv-idseecctoinotrnolfrtoimsswuheiscfhoralalntyispsousessibelxeamhiisnteodpaatthonleocgricoaplsyevwaillluabteiornestaoifnegdr,osins lesions). Unless specifically cited below, all other tissues will be discarded. Blood and processing) Tissue Sample Collection (See ATTACHMENT for tissue collection and Cbeafposrean(edmpltabye)laedndtuabfetesrwrielltebnetiwoeniogfhseadm(pcloemsbfionresduwbesiegqhute,ntto utsheenienaprheasrtma.c0o01kignreatmi)c awneailgyhstessw.illTbheesiencwleuidgehdtswiwtihlltbhee pdaocckuimnegnltisetdpriinorthteo srhaiwpmdaetnat.. aSnadmcpolpeietsubofesthweilslebe. labeled sample widietnhtitfhiecasttiuodnyanndumcboelrle,ctainoinmtailmeipdoeinntti.fication, date of collection, study day, Female Rats Assigned to Caesarean-Sectioning: Oeinghdtaryat2s1deofsipgrneastuedmefdorgCeasetastairoena,nb-lsoeocdtiaonndinlidveorssaagmeplgerosuwpilsl be collected from 110 7, 12 and 13. the The time of sample collections will be recorded in the raw data. cBalvooadasnadmspelpeasra(attedleianstto 4twmoLaepapcrho)xiwmialltebleyceoqlulaecltealdiqfurootsm. thTehreatfsirvstiaaltihqeuoitnfweirlilobrev. ena spelpaacreadtionrtotuEbDeTs.A-Tchoaetseadmtpulbeesswailnldbethsepsuencionnad caelnitquroitfuwgiellabnedttrhaensrfeesrurletdinigntsoersuemrum p(odilvyipdreodpyinlteontewtoubaelsi.quoAltls)s/apmlpalsmeas (wiolnlebaeliiqmumoet)diwailtleblye ftrroaznesnfeorrneddriyntiocelaabnedlemdaintained frozen (<-20C) until shipment for analysis. Fwoelilgohwtiwnigllcbolelercetcioorndoefd.blAoodporstaimopnleofs,eatchhe lliivveerr osfameapclhe (rartigwhitlllabteeraelxcliosbee)d wainlldbtehefllaisvher frozen in analysis. liquid The mneidtrioagnenliavenrdlmobaeinwtilalibneedffrroozzeenna(n<d-7s0toCr)edun(t-il20shiCp)meunnttil for possible. shipment for alnivaelryswiilsl. beAfsiexcetdiionng(lauptperroaxlidmeahtyedleyf0o.r3po10ss0i.b5lecmfutwuirdee)anfarloymsitshbeyreelmeacitnrionngmpiocrrtoisoncoopfy.the `The remaining analysis. portion of the liver wil be frozen and stored (<-20C) until shipment for Female Rats Assigned to Natural Delivery: Blood. milk, postpartum. hTeiarmteaonfdsalimvperlseacmopllleecstiwoilnls b(eblcoooldleacntdedmfilrko)m wmilal tbeemarelcorartdsedonindtahye5raw data. 418-018:PAGE C-17 Protocol 4P1a8g.e01187 Ohonusdaeyd 5forpoasptppraorxtiumma,teelaychfodurahmouwrisl.l beThreemdoavmedwilflrbome tihnejencetsedtiinngtrbaovxenaonudsliyndwiivtihduoanlley sunaimtpolfe)oxayrteoccionllaepcptredo.ximTahteelsyafmipvleemsinwiultlebsebiemfmoerdeimaitleklysafmrpolzeenso(natdlreyasitce10a0ndpL. per maintained frozen (<-20C) until shipment for analysis. fFrololmotwhiengramtislkvisaatmhpelienfceorliloerctvieonn,a bclaovoadasnadmpsleepsar(aatteldeaisntto4tmwLo aepapcrho)xiwimlaltbeelycoelqlueaclted aliquots. The first will be transferred aliquot will into serum be placed separator into EDTA-coated tubes tubes. The samples will and the be spun second aliquot in a centrifuge and the resulting serum (divided into two aliquots)/plasma (one aliquot) will be transferred dry ice and into labeled maintained fproolzyepnro(p=y-l2e0neC)tuubnteisl. shAlilpsmeanmtplfeors awnilallybseisi.mmediately frozen on Following collection of milk and blood samples, the heart and iver of each dam will be. collected. The liver weight lobe) will be flash frozen in will be recorded. A liquid nitrogen and portion of each liver sample (right maintained frozen (<-70C) until lateral shipment (5-20C) for possible analysis. The until shipment for analysis. median liver lobe The heart will be will be frozen and stored excised and two cuts will be smuardfaecetoaatltlhoew approepxearnfdixaetxiotne.ndTahnetefriirostrlcyutanwdillvesntatrrtalolytthoetrhieghptuolfmtohnearveyntarrtaelrym.idTlihnee second cut and extend will be made starting to the through the left ventricle to left the of the ventral midiine surface at ascending aorta. The cut heart the and apex a section fixed in (approximately gluteraldehyde 0.3 for 10 0.5 cm wide) from the remaining possible future analysis by electron portion of the liver microscopy andlor will be. light `microscopy. shipment for The remaining analysis. portion of the liver will be frozen and stored (z-20C) until `Sample Analysis and Shipping Instructions: Appropriate be shipped samples frozen on will dry biecemvaiianotvaeirnneidghftromzaeiln. until shipment for analysis. Samples will One of median each pair of lobes), and serum samples. frozen liver and heart samples matemal liver samples fixed in gluteraldehyde (right lateral will be sent and to Dave Ehresman, packing list awtillthbeepirnecvliuoduesdlywicitthetdhaeddsraemspsl.esTsheentsatompDlaevetuEbheraensdmalni.ver weights and a 418-018PAGE C-18 Protocol 4Pa1g8e01188 Tanhde trreimglayicneirnidgess.erMuimkasnadmplilveersswailmlpbleesanwialllybzeedanfaorlytozteadl fchoorlLesDtLe,roHl.DL,Thteotsael cshaomlpeslteesrol wil be shipped to Or. Saroj R. Das Ani Lyics, 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 shipment recipients and the Study Monitor wil be notified in advance of sample Scheduled Sacrifice - Female Rats Assianed to Caesarean-Sectioning: Oannddaaygr2o1ssofnepcrreospusmyedofgtehsetatthioorna,cifce,maabldeomraitnsawlillanbde psealcvriicfivciesd,ceCraaswsiallrebaen-pseercftoirmoende.d, dBelsocordibaedn.d tiUtsesruieosfaamppplaersen(tbllyoondonsperreugmn/apnltasramtasawnildlblievesrt)awiinlledbewictohll1e0ct%edamasmoprneivuiomusly s1u0lf%idfeortomacloinnffiorrmptohsesiabblseefnucteuroefeivamlpulaatnitoant.ion sites" and retained in neutral buffered `Scheduled Sacrifice - Female Rats Assigned to Natural Delivery: tOhnoradcaiyc,5aobfdloamcitantailon,anfdempaellveicravtisswcielrlabewilslabcreifpiecrefdoramnedd.a gBrloososdnaencdrotpissysuoefstahmeples. (blood serum/plasma, heart and liver) will be collected as previously described. `Reaxtasmtihnaetddfoorngortodseslilveesrioansl.itteBrlwoiolldbaensdactriisfsiuceedsaomnpdlaeys 2wi5llonfoptrbeesucomleledctgeeds.tatUitoerniawnildl baendstraeitnaeidnewditihn 10% ammonium neutral buffered sulfide to confirm the absence of implantation 10% formalin for possible future evaluation. sites" 418-018:PAGE C-19 Dams with No Surviving Pups: Protocol 4P1a8g.e01189 oDrapmrseswiutmhendocasnunrivbiavliinzgedp.upsBlwoiloldbaensdactriisfsiuceedsaafmtpelretshe(blalsotodpusperiusmf/opulnadsmdaea,dh,eamritssainndg liver) will be collected abdominal and pelvic as previously described. viscera will be performed. A gross necropsy Postpartum data of the thoracic, 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 `meaxdaem.ineTdheforratthsewicllaubseeeoxfadmeiantehdofrormogrriobssunldescioonnsd.itiWonheonn tphoessdiabyleth(enootbpsreercvlatuidoend ibsy 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 10 the extent possible. Uteri of apparently nonpregnant rats will be stained with 10% `ammonium sulfide to confirm the absence of implantation sites" and retained in neutral buffered 10% formalin for possible future evaluation. TESTS, ANALYSES AND MEASUREMENTS - F1 GENERATION: Viability: Postpartum Period Litters wil be observed for dead pups at least twice daily. The pups in each liter wil 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 appropriate by the Study Director and/or the fSrteuqduyenMtolnyittohra.n cited above, if deemed Body Weights: Postpartum Period Days 1 (birth), 2, 3, 4 and 5 postpartum. Pooled litter `weights wil 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. 418:018:PAGE C-20 Protocol P4a18g.e02108 F1 Generation Pups Found Dead on Day 1 Postpartum: Pstuaptsusthaattbidrithe. beTfhoereleuxnagmsiwnilaltiboenroefmtohveelditaerndforimpmueprvsieabdiliintywwaitlelr.bePeuvpalsuwaittehdlfuorngvsittalhat sainndk twoillhabveeiddeinetdifsiheodratsysatfitlelrbobrinr;thp.upPsupwisthwiltuhnggrsotshsatlefsloiaotnswiwllilbebeidpernteisfeiredveadsilniBvoeubionm',s Solution for possible future evaluation. Should postmortem autolysis preclude these evaluations, it wil be noted in the necropsy data. F1 Generation Pups Found Dead or Moribund on Days 2 to Postpartum: Pleuspisonfsoaunndd fdoeratdheorcasaucsreifoifcetdhedumeortiobmuonrdicbounndditcioonndiotriodneatwihl.l bPeuepxsamwiitnhegdrofosrsglreossisons fevoaulnudatoinond;agyrso2sstole4sipoonsstopfaprtuupmswfiollunbde opnredsaeyrv5edpoisntBpoauritnu'ms wsiolllubtieonprfeorseprovsesdibilnenfeuutturrael buffered 10% formalin. Should postmortem autolysis preclude these evaluations it wil be noted in the necropsy data. Scheduled Sacrifice - F1 Generation Pups (See ATTACHMENT 4 for tissue collection and processing) Olensidoansy w5illpobsetpparretsuemr,vepdupinsnweilultrbael sbaucfrfiefriecded1a0n%d feoxramamliinn.edNfeorcrgorposssy lweilslioinnsc.luGderoass seixnagmliencartoisosn-soefctthieoncroofstsh-esehcetaidonaetdtbhreailnevfeolroafptphaerefrnotnthayld-rpaorcieepthaallsyu.ture and Cbeafposrean(edmpltaybe)laedndtuabfetesrwrileltebnetiwoeniogfhseadm(pcloemsbfionresduwbesiegqhtu,entto uthsee nienaprheasrtma.0c0o1kignreatmi)c analyses. These weights will be weights will be included with the documented in the packing list prior to raw data, shipment. and copies of these Sample tubes will be labeled with the study number, animal identification, date of collection, study day, sample identification and collection timepoint. Blilttoeor)dasnadmpslepeasrwaitlledbeinctoolfloeucrte(dtwvoiapcearrsdeixa)c appupnrcotxuirmeaftreolmy eeqaucahlpaulpiq,uoptoso.leTdh(ebyfirssetx per aliquot will be per sex wil transferred be into placed serum isnetpoaEraDtToAr-tcuobaest.edTthuebessaamnpdletshewilslebceonsdpualniqiunoat per sex centrifuge. and the resulting serum transferred into labeled (divided into two aliquots)/plasma polypropylene tubes. All samples (one aliquot) will be. wil be immediately frozen on dry ice and maintained frozen (<-20C) until shipment for analysis. 418-018:PAGE C21 Protocol 4P1a8g.e03281, wTehieghltiverrecforrodmede.acOhnpeupliwvielrlfbreomcoelalecchteldi,ttepropooloeldwi(lblybseexrepmeorvleitdera)nadndfixtehde ipnooled glilvuetrserianledaehcyhdeliftoerr ppoooslsiwbillle bfeutudrieviadneadlyisntiostbhyreeelescatmropnlemsicorfosecqoupayl.nuTmhbeerre(mawihneinng possible). analysis. Two of the samples will be frozen The remaining sample will be flash and stored (<-20C) until shipment for frozen in liquid nitrogen and maintained fthreozfeinrst(t<-w7o0maC)leunatinldsthwiopmfeenmtalfeorppuopsssifbrloemaneaalycshisl.iteTr h(epohoelaerdtsbywisllexbepecrollliettcetr)edanfdrowmill lbieghftimxiedcrionsgcolpuyt.eraTlhdeehyrdeemafionripnogssciabrlceafsusteusrewiallnableysdiissbcyaredleedctwriotnhomuitcrfourstchoepryevaanlduaotrion `Samples will be shipped for analysis as previously described for dams and fetuses. 418-018:PAGE C-22 Protocol P4a1g8.e02128 PROPOSED STATISTICAL METHODS?! 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 Test" |. Parametric il. Nonparametric" A. Bartlett's Test" A. Kruskal-Wallis Test (5753 ties) Significant at p0.001 Not Significant Significant at p<0.05 Not Significant Nonparametric Analysis of Variance Dunn's Test Significant at p0.05 Not Significant B. Fishers Exact Test (75% ties) Dunnett's Test ll. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilties 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. 418-018:PAGE C-23 Protocol 4P1a8g.e02138 DATA ACQUISITION, VERIFICATION AND STORAGE: 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)] andlor The SAS System (version 6.12). Records will be reviewed by the Study Director and/or appropriate management parecrhsiovnenselofwitthheinTe2s1tidnagyFsacaifltteyr.geAnlelroartiigoinn.al Aaltl aoriwgililnablerbecoourndds awinlldbiendsetxoerde.d iAn cthoepy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facilty 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: TPersottocaonldaCnodntAromlenArdtmicelne,tsV.ehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. TMraetiantgmeHnitsto(irfy.prescribed by Staff Veterinarian). GCleinneicraallOCbosmermveanttiso.ns and/or General Appearance. Pharmacokinetic Sample Collection, Processing and Shipment. Body Weights. Feed Consumption Values. Caesarean-Sectioning Observations. Natural Delivery Observations. Litter Observations. Gross Necropsy Observations. Organ Weights. Photographs (if required). Study Maintenance (room and environmental records). Feed, Water and Bedding Analyses. Packing and/or Shipment Lists. 413-018:PAGE C-24 Protocol P6a1g8.e02148 KEY PERSONNEL: Executive Director of Research: Mildred S. Christian, Ph.D. Fellow, ATS Directofr 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 Directorof Laboratory Operations: John F. Bamett, 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.., ACVP FINAL REPORT A comprehensive drat 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. EAvpapleunadtiicoens:of TFiegsutreRse,sulStus.mmary 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 drat 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 Facilty'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 altemative (in vitro) procedures were available for meeting the stated purposes of the study. 418-018:PAGE C-25 Protocol 4Pa1g8e01285 REFERENCES: 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, Novembe7r, 1983), J. Ciin. Psychiat. 45(9):7-10, 3. Lang, P.L.(1988). EmbarndyFetoal Developmental Toxicity (Teratology) Control Data in the Charles River Cr: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 of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski,E. (1964). Farbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol, 247:367. 6. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, lowa State University Press, Ames, pp. 240-241. 7. Sokal. RR. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, WH. Freeman and Co., San Francisco, pp. 370-371 8 Snedecor, GW. 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). Amultiple comparison procedurefor comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-1121. 10. Sokal, ALR. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, WH. Freeman and Co.. San Francisco, pp. 388-389. 11. Dunn. O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241252 12. Siegel. S. (1956). Nonparametric Statistics for the Behavioral Sciences, McGraw-Hill New York. pp. 96-104 PROTOCOL APPROVAL: FOR THE TESTING FACILITY anled AGdoorsggoE.tie DDieraercltoovreo,f PRhe.sDe.a,rDcAhBT 2 4 DT nd G. , Ph.D. \DABT Assdtiate Directorof Research Study Director 418-018:PAGE C-26 [----fen UAL... Date BIAGIO Date Dena C. Lebo, V.M.D. Chairperson, Institutional Animal Care and Use Committee FOR THE SPONSOR Missal Godel]... DSteuadnynaMoJn.itLourebker, M.S. Johnid CZ John Butenhoff, Ph.D., DABT, CIH Altemate Study Monitor Date LSlaa/ao....... Date se pun 2000 Date 418-018:PAGE C27 ATTACHMENT 1 STUDY SCHEDULE ATTACHMENT 1 418-018:PAGE C28 ProtocPoalg4e181.001128 SCHEDULE REPLICATE 1 22 AUG 00 Animal Receipt - Acclimation Begins (Female Rats). 29 AUG 00 - 12 NOV 00 29 AUG 00 - 20 NOV 00 Dosage Period - Female Rats Assigned to Caesarean-Sectioning (42 days before cohabitation and continuing through ay 20 of DproessaugmeePdergeisotdat-ioFne)m.ale Rats Assigned to Natural Delivery [42 days before cohabitation through day 24 of presumed gestation (rats that do not delivera litter) or day 4 postpartum (rats that delivera litter). Cohabitation Period (Maximum of 14 days). 090CT00PM-160CT 00 AM Male 1 (07 days) 160CT 00 PM-23 OCT 00AM Male 2 (07 days) 100CT 00 230CT 00 First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. 310CT 00 13 NOV 00 First Possible Day 21 of Presumed Gestation Caesarean-sectioning, Last Possible Day 21 of Presumed Gestation Caesarean-sectioning, 310CT 00 17 NOV 00 First Possible Delivery (Day 21 of presumed gestation). Last Possible Delivery (Day 25 of presumed gestation). 04 NOV 00 17 NOV 00 First Possible Day 25 of Presumed Gestation Female Sacrifice. Last Possible Day 25 of Presumed Gestation Female Sacrifice. a. The study initiation date is the day the Study Director signs the protocol. b. Replicate 2 will be initiated two weeks after the start of Replicate 1 (see Page 2 of ATTACHMENT 1) ATTACHMENT 1 418-018:PAGE C-29 ProtocPoalg4e182.001128 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 00 - 06 DEC 00 Dosage Period - Female Rats Assigned to Natural Delivery [42 days before cohabitation through day 24 of presumed gestation (rats that do not delivera litter) or day 4 postpartum (rats that deliver a litter) Cohabitation Period (Maximum of 14 days). 250CT00PM---01NOVOOAM Male 1 (07 days) 01NOV 00 PM-08NOV OO AM 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 Oraft Final Report. 418-018:PAGE C-30 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 413-018:PAGE C-31 DATA SHEET S3tM. CePanutle,r Minnesota 55144-1000 1-800-364-3577 or (612) 737-6501 (24 hours) Copyright, 1998, Minnesota Mining and Manufacturing Company. iALnLforrsiagthitson rfeosrervtehde. purCpoopsyeingof apnrdo/poerrldyowuntlioladiiznigng of3M thpirsoducts 11)s tahlelowiendforpmraotviiodnedisthacto:pied in full with no changes unless 2) npreiiotrheragrteheemecnotpyinsorobttaheineodrifgrionmal3Mi,s arnedsold or otherwise distributed with the intention of earning a profit thereon. TDIRVAIDSEIONNA:ME:3M CHEMICALS 10FCN-U9M5BERF/LUU.OPR.ACD.:Brand Fluorochemical Surfactant 9988--00221017--00818083--57 0000--5511113355--0099035642--17 9988..00221017--30911064.-15 ISSZUFEoD0:00J2a-n1u0a4r4y1 29, 1998 oo. SDOUCPUEMRESNETD:ES:10N-3o7v9e6m-b9e.r 05, 1997 0000.-5511113355.-0029301515--28 1. INGREDIENT C.A.S. NO. PERCENT PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SULFONATE...... SULFONATE...... 2795.39-3 62 3871.99.63 86 8 PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SULFORATE.. SULFONATE...... 29420.49-3 60270-55.5 3 2 -7 6 POTASSIUM PERFLUOROALKYL SULFONATE...... 3872.25.1 1 3 2. PHYSICAL DATA BVAOPIOLRINGPPROEISNST:U.R.E.:.............................. N/A NIA VEAVPAOPRODREATNISOINTYR:A.TE.:............................. NIA N/A SSOPLEUCBIIFLICITYGRAINVIWTAYT:E.R.:....................... scla.igh0t.6 Waters PERCENT VOLATILE: .............. (Bulk) 0% BH: ctentannnnnnn7n2i8n VISCOSITY: ...............cccce. (0.1% NID) Aqueous) MELTING POINT:..........0 00000 wip APPEARANCE AND ODOR: Lignt colored, free flowing powder. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately MJSaDnSu:aryFC-299,5 F19L9U8ORAD Brand Fluorochemical Surfactant 418-018:PAGE C-32 PAGE 2 3. FIRE AND EXPLOSION HAZARD DATA FFLLAAMSMHAPBOLIENTL:I.M.I.T.S..=..L.E.L.:................ Nome N/A AFULTAOMIMGANBILTEIOLNIMTIETMSPE-RAUTEULR:E.:............. NNI/AA EXWTaItNeGrU,ISHCIaNrGbonMEDdIiAo:xide, Dry chemical, Foam SPEWCeIarALfuFlIlREprFoItGeHcTtIiNvGePcRlOoCtEhDiUnRgE,S: including helmet, self-contained, apnodsiptainvtes,prbesasnudrsearorounpdresarsmusr,e wdaeimsatndanbdrelaetghsi,ngfaacpeparmaatsuk,s, anbdunker coat protective covering for exposed areas of the head. UNUSSeUeALHazFaIRrEdouAsNDDeEcXoPLmOpSoIsOiNtioHnAZAsReDcSt:ion for products of combustion. 4. REACTIVITY DATA STABILITY: Stable INNCoOtMPaApTpIlBiIcLaIbTlYe.- MATERIALS/CONDITIONS TO AVOID: HAZARDOUS POLYMERIZATION: Hazardous polymerization will not occur HAZCAaRrDbOonUSMoDnEoCxOiMdPeOSIaTndIONCarPbRoOnDUCDTiSo:xide, Oxides of Sulfur, Hydrogen Fluoride, Toxic Vapors, Gases or Particulates 5. ENVIROWMENTAL INFORMATION SPOILbLserRvESePOpNrSeEc:autions from other sections. Vacuum, use Wet sweeping cbeompaonunidgniortiownatesrourtcoe.avoiCdleadnustuipngr.esiCdAuUeTIOwNi!thAwavtaecru.um CPlleaacneerincoanuld approved metal container. Seal the container. RECDOoMMnEotNDErDeleDaIsSePOStAoL:waterways or sewer. Do not use in products or p1r/1o0cesosfesthethaltowecsotuldEC5r0esuolrt LCinS0 aqcuoantciecntrcaotnicoenn.tratIinocnisnergarteeateirn than an industrial material. Coormbcuosmtmieornciaplrofdauccitlsitwyillin inthceludperesHFe.nce Diofspoa saclombustible alternative: Dispose of waste product in a facility permitted to Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately MSDS: FC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 413-018:PAGE C-33 PAGE 3 5. ENVIRONMENTAL INFORMATION (continued) accept chemical waste. ENVIRONMENTAL DATA: 96-Hr. Aquatic Fish LCSO, Fathead Minnow(Pisephales promelas)=38 mg/l, Bgaliuredgnielrli)S=u1n1fismhg(/lL;epo4n8i-sHr.macECrSoOc,hirDuasp)h=n6i8a Mmag/gln,a R=ai5n0bomwg/lT;rouCtO(DS=a.l0m0o4 0/0; BOD20 = Nil. REGVUolLaAtTiOlReY OIrNgFaOnRiMcATICOoNm:pounds: N/A. VOC Less H20 & Exempt Solvents: N/A. Sbienfcoereredigsuploastailo.ns vU.aSr.y, EcPoAnsHualztardaopupslicWaabsltee rNeugmublearti=onsNoomre a(uNtothorU.iSt.ies EPA Hazardous). TTShCiAs, prEoINdEuCcSt, coCDmSpLl,iesAICWSi,thMItThIe chemical registration and Korea. requirements of EPCRA FIRE HHAAZZAARRDD: CLNAoSS:PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes 6. SUGGESTED FIRST AID EYE CONTACT: Iamediately flush eyes with large amounts of water for at least 15 minutes. Get immediate medical attention SKIN CONTACT: Inmediately flush skin With large amounts of water. Remove contaminated contaminated ccllootthhiinngg. beIfforierrirteuastei.on persists, call a physician. Wash INHALATION: If signs/symptons occur, remove person to fresh air. If signs/sysptons continue, call a physician. IFDrSiWnAkLLOtWwEoD:glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION EYAEvoPiRdOTEeCyTeIOcNo:ntact. Wear vented goggles. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately JMSaDnSu:aryFC-299,5 F19L9U8ORAD Brand Fluorochemical Surfactant 7. PRECAUTIONARY INFORWATION (continued) 418-018:PAGE C-34 Pace 4 SK`IANvoPidROTsEkCiTnIOcNo:ntact. material. A pair of Wgelaorvesappmradoeprifartoem gthleovefsolwlhoewninghanmdaltienrigalt(hsi)s are pZFoeevrcesorominmneagnl,depdcr:oovteercbatultilyosln. riutbePbmresort.easctiUnvseeecesgosnaearrmyeonrttsomorp(eroetvohefenrtthtehsaknifnolgllcooowvnietnsag)ct:shouhledad pbeolmyaedtehylofenee/iptohelryvionfyltihedenfeollcohwlionrgidemat(eSrairaalnse:x) REUVCseOenMtEiWNliDatEthDedaVpEpaNrrTeoaIp.LrAiTaItPOerNo:vliodcealsuefxfhaiucsitentvenvteinltaitliaotni.on UtsoemainintaaiWnell: e15mishsotionasdeqbuealtoew, reucseommaepnpdreodpreixaptoesurreespilrimaittosr.y prIfoteecxthiaouns.t ventilation RE`SPAPevIsoRpuAidrTaObtRroYerasPtRhOibnTagEsCeTdoIfOoNnd:usati.rboSrenleecctonconeentrofatitohne fofollcoownitnagminNaInOtSsH aapnpdroivned GRcaclofrmdaasnkceswuiptphlieOdSWAairregruelsaptiiroantso:r, fhualllf--fmaacsek dduusstt aanndd mmiisstt rreessppiirraattoorr,, full face supplied air respirator. PREaDVoreEaNnsToItOtNehaotOr,FouAgdChrCliyInDkEMNioTrtAhLssmooIakNpeGESwaTnhIdeOnNw:autseirn.g thWiasshprhoadnudcst.afHtaershheaxnpdolsiendg and before eating. RECKOeMeMpENcDoEnDtaSiTnOeRrAGdEr:y. Keep container closed when not in use. FIRNEonfAlNaDsnEaXbPlLeO.SION AVOIDANCE: OTHNCEooRntsaPmmRokiEinCnaAgtU:iToInOSNmAooRfkYintghIeNFwOhtRioMlbAeaTcIcuOosNi:nagnd/tohris sparookdeucatnd calneadrestuoltthein formation GtfhastheMSDnSa.zardous decomposition products mentioned in section 4 of HHS HAZARD RATINGS: HPEEARLSTOHN:AL2PROFTLEACMTMIAOBNI:LITXY:(Se0e RpErAeCcTaIuVtIiToYn:s, 0section 7.) EXPOSURE LIMITS INGREDIENT VALUE UNIT PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SSUULLFFOONNAATTEE...... POTASSIUM PERFLUOROALKYL SULFONATE... 00..11 ~ MMGG//MM33 0.1 MG/M POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 Avbreviations: NID - Not Determined N/A - Not Applicable TYPE AUTH SKIN TTWWAA 33%M TTWwAa 3M M Y CA- Approximately MJSaDnSu:aryFC-299,5 F19L9U8ORAD Brand Fluorochemical Surfactant 418-018:PAGE C-35 PAGE 5 EXPOSURE LIMITS (continued) INGREDIENT VALUE UNIT TYPE AUTH SKIN POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 TAM t=heSKIpNotNeOnTtAiTaIlONc:ontLriisbtuetdionsubtsotatnhceesoveirnadlilcateexdposwuirteh b'yY' thuendecrutaSnKeINousrefreoruteto ibynclduidriencgt mcuocnotuasctmewimtbhranteheansdubsetyae,ncee.itheVrehibycleasirbcaonrnealotre,r msokrien apbasrotripctuiloanr.ly, S"OaUR:CE OFaMEXRPeOScUoRmEmenLdIeMdITEDxApToAs:ure Guidelines 8. HEALTH HAZARD DATA EYEMilCdONTEAyCeT:Irritation: signs/sysptoms can include redness, swelling, pain, and tearing. SKIMNildCONSTkAiCnT:Irritation (after prolonged or repeated contact): signs/sysptons can include redness, swelling, and itching. Meaxytenbdeedabstoimreb.ed through the skin and persist in the body for an INHMAaLyATbIeONh:araful if inhaled. May be absorbed by inhalation and persist in the body for an extended time. Single overexposure, above recommended guidelines, may cause: sIorrreinteastsionof (tuhpepernosreespainrdattohrryo)a:t, sicgonusgh/isnygaptaonmdssnceaenziinngc.lude IF ISnWgAeLsLtOiWoEnD:is not a likely route of exposure To this product. Illness may result from a single swallowing ofa moderate quantity of this material. May be harmful if swallowed MUTAGENICITY: Mutagenicity assays indicate the product is not mutagenic. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately MJSaDnSu:aryFC-299,5 F1L9U98ORAD Brand Fluorochemical Surfactant 413-018:PAGE C-36 PAGE 6 6. HEALTH HAZARD DATA (continued) REPNRoOtDUtCeTrIaVtEo/gDeEnViEcLOiPnHEtNhTeALratTOXaItNSo:ral doses below maternally toxic Levels. OTHTEhRisHEpArLoTdHuctHAZisARDnotINkFnOoRWMnATItOoN:contain any substances regulated under California Proposition 65. A Product Toxicity Summary Sheet is available. SECTION CHANGE DATES HEADING SECTION CHANGED SINCE November 05, 1987 ISSUE Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately TbheecoirnrfeocrtmatasionofinthethidsateMatiesrsuieadl. Saf3eMtyMAKDEaStaNOShWeAetRRA(NMTSIDESS), isEXbPeRlEiSeSvEeDdORto MIEMRPLCIHEADN,TABIINLCILTUYDINOGR, FBIUTTNESNSOTFOLRIMAITEPDARTTOI,CULAANRY PIUMRPPLOISEED WORARRCAONUTRYSEOFOF PWhEeRtFhOeRrMANtChEe OR 3M UpSrAoGdEuctOFiTsRAfDiEt. forUsearpaisrtirceuslpaornsipbulreposfeoranddetesrumiitnaibnlge for cuasner'asffemcetthotdheofuseuseandorapappplliiccaattiioonn.of aGiv3eMnprtohdeucvta,riestoymeofoffaWchticohrsartenat tuhneiquuesleyr eWivtahliunatethetheusaeMr'sprokdnuocwtledtgoedeatndermcionnterowlh,ethiterisitesissenftiitalforthaat particular purpose and suitable for user's method of use or application. D3Mue prtoovitdhees riemnoftoermaptoisosnibiinliteylectthratonieclecftorroaniacs a service to transfer may its customers. have resulted irneperrersoernst,atioomnisssiaosnstooritsaltceormaptlietoennsesisn torhisacciunrfaocrym.atioInn, ad3dMitmiaokne,s no iinnffoorrmmaattiioonn oinbtathienedMSfDSromavaaildaabtlaebasdeiremcaytlynotfrobme a3sH. current as the ovodw dy vie eit ren 418-018:PAGE C-37 . sroouCcTEs: 8:AOR Rcos0Sq3S0Y wa mFez,MtCVAalTEide156C0/D2s0LH0N0O-TFLDBaAEIurgLSNso a ETr soknREoess a6rSRo9m3:O54OTIS a S2Sio0g82ni0tS.apd0riu0eciecwnostreeaett03, SEnisial phone: 314 u1s71 5768 FTeoergean0c0y22p5hon$e0:52314 771 5765 aan gcrIon met 1. - - (es <= = = = =o CHEMICAL TOENFIFIGATION- = << << eferCtaazELs +: |... couposcCiehsoooissosn/tIeNaFoORMATION ON INGREDIENTS - = = = = STECEevhoCrhama2s0os0h-a3m8e3s2 SeSE Oo-swAS -E tN-oe 3r-s0tLIiot) uL(3hse-om-8toEBueIeAEs)Tsn=TAioocn(roS-CLI3+)-3CEHASOAC-L-HZC0OSHLTLOEELRSEYTSL-TCSER-NOAT-LLN3EC-S-O3TBH-SEORBTLIEANT-+AO-L*DOYLFCHHOO=LL:6E=STE3R-INE scER rAEE SOAROGKYICHHOOLLEETST S8 -EETNEE + pnPtRaORVDIsTANTIONENDTIFICATION = = = = = = 7 - < SrcRITnONTToAEJaRNOOFTToRoAFAVVAAATcTToLILNAAAEBRCALEEsEE, |15IMWMMVEIEDN.DIUIAeTATErTESmE.LLsYYmoWFaLAmUSdSHHwSmEKAYIsENSuRWmWIsIT-THH-SCOOAoPPoIOAUN-SD<AC=MOOPUI-NOTU-SS -OF JA iRoOtUooUNNTTErSs ONo.FF WcAvWTAiTTEiaERcS.aiseR,ezaoTHITNRGsHIsAIDn.IFFIICfULNTO,T 00s MOUTH WITH WATER SGPRIREVOEAVTIHDOIEXNDYGGEPNGE.IRVSEONARITSIFCIOCNSICAILOUS. SeRhBEiSLnlRRAovpO mivaSMAIICNIADATTNiZa.DehO CLOTHIE NG SEFOFRIEGHTRIENUGSE.MEASURES = = = = = = = = = SxGTEAozNeEGRUISSHTiIRoNAiGYoMsE,DIARr CHEMICAL PONDER OR APPROPRIATE FORM. SeESCRTEAvLENE:FIRCNSOENITGGACHTII,NWGITHPBROASCCKEAIDTNUHRIAENNSGD AEPYEPSA.RATUS AND PROTECTIVE CLOTHING 10 eCNeeUSiUnAsL TFoIxREicANFDovgEsXPLGONSDIEORNOSFIIRHNLAZTACARODNLSDITRIEOLNESA.SE MEASURES- - <= - = = = = oPSRmnweitIiRaSorrr:zseanBriasvnecyCGLIoNOgTaAEiNi3Gsa,6 AGNLDOVEHSOLDANDFORMASWKA.STE DISPOSAL. SoRBiomOt>OASmtsacceaA5UaSnTs WASHSISPTILALNDSLIITNEGAFATNDERSMTAOTREARGIEA-L-PI=CK=UP- I=S=CO~M~PLETE. seeSSreHrBseun.Iitaol72 SsSeaEaFTrsIiToYnGO62.GGyLpEoSs.une conTRoLs/ PERSONAL PROTECTION- - - = = = STCOoMSPhA/TMIBHLnE-ACPHPEAMOIVAELD-RREESSIPSITRAANTOTR.GLOVES. Tage JSR 418.018PAGE C38 propuCCTCT+:,A C850S3n00S17H//E22E85T//,2200V0N0aA00lMiE1d:185:8C:/0H02O330L0E0ST-ERMiaasetktosimsesuaepsrsRavataAssnebs 6e3ra91Ts2rT51ae45a7z:smn -- Somes AN 5 vegan SRAEVoCOlHToOANICICONANHLTALAACETTXIHOAWNUI.STTH OR REQUIRED. REYEEPSE,ATESDKINCXAPNODSORCELOTHING. AYkVeoGei?hTTIHPGORHROTOLLOUNYGGHELCDYLOSAEFDT.ER HANDLING. se`rAiPToPoEnmAReANICNETRAANDCCO00OOLDOORRDYRY PPiLlACCEi.car awo chemical sROPEATIES =< PHW`YHsSoIIzTCiEAiLnPGOPWRPDOoEzPRNEtR:TIES MELTING POINT: rc360To Ms C ronSaineiIricScIRaTviYTy: 2.0s6m7mernrny aw meAcTIvETY - = mmc o CC StaSptIaLsIiT.Y I{NACSSOTAMRROPONAUGTSIBOIXCLIOIDMTIBIZUEISSNTGIONAGEONRTSDECOMPOSITION PRODUCTS HATZZoAAxRRiBDcOONUSroMvOFeNOsOLXYIoNDFEE:R,:ZACTARISOONN DIOKIDE eerWiILLLTONTOTOOCCZUR.ER. 2oxzcoloSIAD INFORMATION - = = = << << acuMErAsYECzArUrSeEEemtAswIrRURLITABYTIOINN.HALATION, Re en: Sia INDICATE A INGESTION, OW ORDER OR SKIN ABSORPTION. OF TOKICITY. GENERAL GOOD RTEOCUSST8R:EEFP2I8N4G000P0R0ACTICES SHOULD 3 FOLLOWED. TARCRNGOhiEO7oTNiuEcsYrOT,sREGRAUOONRNLETDFEAERTR,ATILBILTAYDDE(RPOS(TS-LIAMDDPELRANTTAUMTOIROSN) MORTALITY) moSFToreRiTicrroistcuOiNDcEVFEE(LRFOTQPIULWIIEVTNOYTCAALL(LATIBUTNNTOEORRRMIAGSLEIZINETI)ICESAGE(NCTRABNYIOFRATCEICASL)CRITERIA) oTTONrMeyOicRoIS)GSELNiSIehCtieaD(TTSRUEMOGPRIRSSZTSRAETYNTOESFIDTETHEORKOEFI.CAPCSPEFLEFIECCAATCTSTIUOAONFL) CEHNETMRIYCAILN SRUTBECSSTANFCOERS SectiCDoOAnMTPAL13EN.TOET mIYnENTFOeRAeMVAAlTIILOAoNBl.LE. sccromn iso MATTEL | 5COLOGICAL INFORMATION = = = t= t= = <= prssosas constomaaTIons - - = - =< - - CHOHE%SNsTGuAVnvELEAoLInLNCNIFINEXDEERRATATHLOE,RVPAESTTQTSUAERITIEZAPMLEApDNaDWnIWTsILHTOsHCoAArLAtNCIOAMNIFBVNUTIFSEROTRORIBNMBUMALRETENNIETORANSLOAL-NVRDEE=NGTUS-ICAAR-NTUSID=SOENB=RSU..R=N IN = A sscrtIoNnTAnCeTn SECTION 16. SooInoGMmAco<CHo=EMnI2 CA=L - CO- MPRAENGYULAFTORORYTRAINNSFPOORRMTAATTIIOONN = IN= FO=RM=AT=IO=N.= = ~~ REUS$TEATokuceSu,aCkANSCTEARNDARREDVSI,ZW:ANADNIMRAELGULIANTAIDOENQSUATE EVIDENCE IMEMDT 10,99,1876 we T are coR mmitted 10 tne ST uceusrasgeA of a2us CuR riamers. WEmAplasTess and se 0 i sn 0 vrs run duct #: C8503 wn wn wan 2 413:018:PAGE C-39 NAME: CHOLESTEROLSLOoSvDteIMLato CE10L3 UTSA o Pro ICTR NtGn070S7/H/2E2E55T//,2200V00a00li1d815::/00233--000 - WTTEReinmiresospseaosmssaesmnenT7787 -- - =r IARC . iis CAANNCCERERBRbEVIaEosWe:sG;ROUn?s3 8: TF = 1321 Nos 10; ZuTTsNEuEDLG137L0,0A163,r1e99730220 INOZAEEHRSASC wCC1e5h3r41eSEWCeTPbIROONsGoRsA8eM(sB;)198NC8iH,sEMIN5;CEGAALTTNIFIVNEV:0ERsNEiTSA;OHLRETYw-RoC/sLSOAN1FAE;LTYASSSTAUYDIES TERR periih EETCSORRT{ERQScTUrrsoZwniR8sI(spI)ioUNNPU(BTLSICnSAuHTmSE)Da uaTioN IS BELIEVED DTnA0sToA8n5aBAtCSIOERO,RNE:CDTECEBMUBT7ER00ne81s8393NsoOrLALDrSRuuInRCeeHoO,nRsT TO TTSFOEELAKUKAA1%.SeiHReACsLiLIUuNSpNIoFoVmrE BWASeNnDWnELaSDHzALLLAIbSBoL5vEeUSOsERRDoouAOcNNTLY.Y DAANSSMDEAEIGATEIRGOEUNRVISEEDRSES.UOELFTISSSN1ALG0LOEF.WFLOOFM IHWAUNSDRLCIENGor OOEREGvRC OIoH?ou1s933FoRS1I6AGDMDAWIa-TkAeILODNROAINLCLHITWEICORT.MESD AND CO PASER COPIES FOR INIERAL US2 GNI wereericrmoiimemesethrehseh LSiuecepSraenrgieenhce.s3urTeCcuhnntaamieargsy. anFdupSleorsveiecse ond 418-018:PAGE C40 J 5 Material SafeOtuytOuDUtnaaPatrreaessS017h120e5121e0509t09 Viren 19 Section 1- Product and Company Information -- PPrroodduucctt NNaambeer Band CStormeeptaAndydress TC=ietty.hSmentParZoin,teC:ouent,ry aOLeMsE?VALONEC ACID LACTONE Sam cremea S30m50aSnuacenSuet 2SSt0oa0vr3r2a5s0NrO5e.2s 63102.Us Emergency Phone: "raza assns Section 2- Composition/nformation on Ingredient SuBbUsNtEanVcAeIOaNmEeAci LaCToNE ecarsaes FSoyrmmuolans cono0a sarNoasy Section 3 Hazards Identification Foraastona marmaron on xy. seas refer o Secon 11 Section 4- Firat Ald Measures onal Eoxpsourweash ok uth wi wats proves pron consis. Cal syscan afiinnns rExepomntrooeesvae. tea Geatcrespraton. Hf eaiinncgut ve oxygen. DIenrcmaaselofExcpoosaucre medately wash skin wh soap nd 3900s amour of var EyCeaEsxpocuortea. mesa sh pes wih 25003 amour of water or tea: 15 meses. Seo ction 5- Fie re Fighting cMeasmureT s --eee ne eee -- Finan Poi: sr me Autoignition Temp: Na Flamm: NA ExtiSnognuaibalneing Media Water sry. Carson dance ry cham) sewer,or sss fou 418-018:PAGE C-41 Fictriogthteicngte Exuipment Wese caontarined being a69aatu and prsecive cong0 rave Cora weh skin and eyes Sec6-tAcicidoentnal Release Measures WPeroacrerduersesp).ofcPrearsmccraallsPareecyauGtoigongt.s) ober oct.47 heyruber aves AMCboesmtpohlroesdtsoenfosraGnedaorfivngeUpc and piace in dosed containers for Sspesal Ventaie aranewaash sp $18 Se: material 9464p Section 7 - Handing and Storage -- HanUdsienrgExposure Avi ition Aviecontactwin eyes, sin, an SOG AVS 0GN0d Bf UBOAIES BPRS Sto"rSaugtesble Keep any cose Section 8- Exposure Controls / PPE ESnagtienreesrhionwgerCoanntaraoyles ban Mechancal exnaus ecurec, PersRoensapilrPartootreyctive Equipment NHIaOnSdHAMSHA approved respon. CECrohemepmaenaiiesscareemscoagtgerse.msare goves GWeansehraclonHtyagieanteeMdecaasuirnegsSeton reuse Wash thorough ate: arcing Secti9on- Physical/Chemical Properties sopearance MolecularWeight CFolaomry sage 130.15AU FSoortdmtes mass or ragmares Broperty valu. oBHPE Range N14a5. 150C MFrePezPingRaPnoginet 2Nac VVaappoorr DPreesseutre NNAA SGSaDtaurnaatetdyVaperCone. NNAA OBdulokreTnhreelsyheld NNAA ATemperao tPruesrsuere Smmig SPamgea2Creme Mage? Sigmsa-Aeldsricsh iCoroponration _ 413-018:PAGE C-42 VVoOiCsCionnt.ent NnAa WSaatveeCoCnotnetretnt NNaA ViEevcapoourtsyion Rate NNAA PDaerciotmipoonsCioteifofnicTieenmtp. NNAA FFilnaasnhPpoomm"CF 2asc AuExopliogsinonomniTteemp NNIAA SaRehfuramctiiyve Index. TNaan `Section 10 - Stability and Reactivity sbi`iCtoyncttioonfs metbilty PCrootnedcit trioomtnmos oAvuoi.d MPraoetctefrotnmomAovositdue Stong oxsizng agers. HazeazrdaorudsoDuescDomepcoosmiptoisotniPornodPurcotdsucts Carbon monde, Carson dons. Section 11 - Toxicological Information Rou"tSekionfCEoxnptoascutre. Moycause wn reason EMyaeyCcoanutsaecetye raven IMnahtaeartaomnay be tang fo mucous mamoranes and user esprairy rac, MMuttspaolerRayomuituelbsyinven, ingestion, o sin assoron, STiogns abnedtSofymouprtkomwsoofnE,xphoseurCehAT BTEC 1GKECIG<aS EBS ave Nt Been OXGUGHY IeSTES RTECS Number: Section 12 Ecological Information Section 13 - Disposal Considerations `lAapprroapriosftemaMxeitohomdaotfeDri)spwoisnsi8 oSf SmubsstanScwaerenr:ep3ara5tio0n 8 PIC InCEBLO EQUZBES WAN 8 ETS! 470 EUBDET bse ander. sae, 3nd aca eveormerts equator Section 14 - Transport Information oorProper Shipping Name: Nore Semsa Creme. wate? Sigma a-Aldri ich Com rporation --_ 418-013:PAGE C-43 NonHazafrordTraonsuporst: Th subsarce icorssered o be ron hazarfororoanuss wParoperShipping Name: None NonHazarfodrAsir Tranaport; Non-hazardous fr a ranssort Section 15- Regulatory Information `USnAitReAd3S1t3atLeisstReedg:uNloatory Information `Section 16 - Other Information ToAWaeirrnaontvneye. rsfaolmtntinbeshbeeldieavieesobfreacroymGeacmabgute6r2e8auriontgaomura0nbdeialofncocmucosratniascstvmwaielhlhbeeussheodveonOyS4Ua GSuedee.ovSegm:s 336 of vCroiimaceodesBuapcekrinogpSiinp ffoorrarciiteornsal5monsy.andconaions of Sale Capyrge 1959 SmaArch Co. Lares Gartea mse Sigma Cremeal Page wise? Sigmee-mAkpimicehsCroripoornation -- 418-018:PAGE C-44 ATTACHMENT 3 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE 418-018:PAGE C45. ATTACHMENT 3 Version: 418.P0r1o8to(c1o6lA4U18G.0001)8 Page 1015 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE CoTnesttroAlrtiActlteicles: ~~ PChFoOleSsterol Vehicle: Mevalonic acid 0.5% Tween 80 in R.O. Deionized Water for PFOS and CRh.oOl.esdteeiroonlized water for Mevalonic Acid A. Purpose: The purpose of this procedure is of dosage formulations of PFOS, to provide a method for the mevalonic acid, cholesterol preparation and the A0r.g5u%sTRweeseeanrch80Lavbeohriactloerfioers,orIanlc.adSmtiundisyt4ra1t8i-o0n18o. rats on Primedica 8. General Information: 1. Aslplecfiofrymutlhaetipornotcoocnotlainnuemrbserwi,lltbesetlaarbtieclleediadenndticfioclaotriocno,deAdr.gusEabcathchlabel will number, concentration, dosage level, preparation date, expiration date and storage conditions. 2a. _SuspenDsiaoyns of PFOSX_will bWeeepkrleypared _ For__ days of use 2. 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: __ paiy X_ Weekly _For__daysofuse 3. Suspensions will be prepared at a final dosage volume of 5 mUkg. 4. SXa_fety:Gloves, lab coat. goggles or safety glasses and faceshield X_ Dust-Mist Respirator __ Halt-Face Respirator -- Ful-Face Respirator/Positve Pressure Hood Tyvek SuitApron 5. Dosage formulations of the test article and control articles adjusted for Free base and % Pury. Yes _X_ No (Calculations based on 100%) T_ FreeBase __ Pury 418-018:PAGE C-46 ATTACHMENT 3 Version: 418:P0r1o8to(c1o6lA4U18G.0001)8 Pago 20's TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE 6. Sampling requirements: Cited in protocol. 7. Storage: Gited 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 bpraerpsaraarteitoon,bealiaqdudotetdintgo, tshaempclonitnaginaenrds/;ormidxoisnaggsehoadumlidniosctcruartidounr.ing 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 aricle (See TEST ARTICLE PREPARATION CALCULATIONS) into an appropriately sized, labeled, pre-calibrated container. QS ad to the required amount with 0.5% Tween 80 and heat the mixture to 80C =5C for approximately 30 minutes or until the TAS dissolves. 2. Obanthc,e atdhed taesmtaagrnteictliechsatsidbiasrsotlovtehde, croenmtoaivneert,heplsaucsepeonnsaiomnagfnreotmitchestiwrater plate and mix until the suspension equilibrates to room temperature. (Be sure there is a visible vortex, this will achieve the desired emulsion.) Be sure that the suspension is mixed prior to and during preparation of other dosage groups, aliquotting, dosage and/or sampling. 3. To prepare the 0.32 mg/mL suspension, remove the required amount of 0.40 mg/mL suspension (See TEST ARTICLE PREPARATION CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ado the final required volume with 0.5% Tween 80 (See TEST ARTICLE PREPARATION CALCULATIONS). Stopperthe graduated cylinder and invert several times to ensure proper mixing. Transfer the suspension to an appropriately sized, labeled container. Add a magnetic 418-018:PAGE C-47 ATTACHMENT 3 Version: $18.P0r1ot8o(c1o6lA4U1G6-0010)8 Page ols TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE sti bar to the container, place on a magnetic stir plate and mix prior to and during sampling. preparation of other dosage groups. aliquotting, dosage and/or 4. T0.o32prmegpa/rmeLtsheus0p.e2n4simogn/m(SLeseuTspEeSnTsiAonR.TIrCeLmEovPeRtEhePAreRqAuiTrIeOdNamount of CALCULATIONS) and add it to an appropriately sized graduated cylinder. QARSTaIdCLtoEtPheREfiPnaAlRrAeqTuIirOeNd CvAolLuCmUeLAwiTtIhO0N.S5)%.TwSteoepnper8t0he(SgereadTuEatSeTd cylinder and invert several times to ensure proper mixing. Transfer the ssiurspbeanrstiootnhteocaonntaaipnperro,prpilaatceelyosnizaedm.aglnabeetliecdsciorntpalianteer.andAdmdixapmriaogrnteotic and during sampling. preparation of other dosage groups, aliquotting, dosage and/or 5. T0.o24prmegpa/rmeLtshues0p.e2n0simogn/m(SLeseuTspEeSnTsiAonR.TIrCeLmEovPeRtEhePAreRqAuiTrIeOd Namount of CALCULATIONS) and add it to an appropriately sized graduated cylinder. QARSTaICdLoEtPheREfiPnaAlRrAeqTuIiOreNd CvAolLuCmUeLAwiTtIhO0N.S5)%. TwSetoepnper8t0he(SgereadTuEatSeTd cylinder and invert several times to ensure proper mixing. Transfer the sstuirspbeanrstiootnhteocaonntaaipnperro,prpilaatceelyosnizaedm,aglnaebetliecd sctoirntpalianteer.andAdmdixapmraiogrnteotic and during preparation of other dosage groups. aliquotting, dosage and/or sampling. 6. T0.o20prmegpa/rmeLtsheus0p.e1n6simogn/m(SLeseuTspEeSnTsiAonR,TIrCeLmoEvPeRtEhePAreRqAuiTrIeOd Namount of CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ad to ARTICLE the final required PREPARATION vCoAlLuCmUeLAwiTtIhO0N.S5)%.TwSetoepnper8t0he(SgereadTuEaSteTd csyulsipnednersiaonndtionavenrtapspervoeprrailatteilmyessitzoede.nsluarbeelperdocpoenrtamiinxeirn.g. AdTrdanasmfaergntehteic asinrd bdaurritongthperecpoanrtaatiinoern,ofploatcheerondoasamgaegngertoiucpss,tiralpilqautoettainngd, mdioxspargieoratnod/or sampling. 7. To prepare the 0.08 mg/mL suspension, remove the required amount of 0.16 mg/mL suspension (See TEST ARTICLE PREPARATION CALCULATIONS) and add it to an appropriately sized graduated cylinder. QS ad to the final required volume with 0.5% Tween 80 (See TEST ARTICLE PREPARATION CALCULATIONS). Stopper the graduated 413-018:PAGE C-48 ATTACHMENT 3 Version: 418P:r0o1to6c(o1l641A8UG0010)8 Page 4015 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 sti bar to the container, place on a magnetic str plate and mix prior to and during aliquotting, dosage and/or sampling. 8. To prepare the 0 mg/m suspension, add the required amount of the appropriate vehicle (R.O. deionized water or 0.5% Tween 80) to an appropriately sized, labeled container (See TEST ARTICLE cPoRntEaPinAeRrA, TpIlaOcNe oCnALaCmUaLgAnTetIiOcNSst)r.pAladtde aanmdamgniextpirciosrttiobaanrdtodutrhieng aliquotting, dosage andor 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. Asidrdbaaprptrootxhiemactoenltyai8ne0r%, opflaRc.eO.ondeaiomnaigzneedtiwcatsetirr tpolattheeacnodntaaignietra.te Aundtidl athe control article has dissolved completely. 3. tThreanfsifnaelrdseosliurteidonvtoolaunmeapwpirtohprRi.Oa.teldyeisoinziezdegdrwaadtuearte(dSceyeliTndEeSrTand Q.S. to ARTICLE/VEHICLE PREPARATION CALCULATIONS). 4. RCeotvuerrntthheegsroalduutaiotnedtoctyhlienodreirgiwniatlhcaongtlaaisnsers.toAppdedraanmdagmniextibcy sitnivrebrasiront.o 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. Wlaebieglhedtchoenrteaqiunierre(dsaemeoTuEnStTofAcRhToIleCsLtEer/oVlEiHnItoCLanE aPpRprEoPprAiRatAeTlIyOsiNzed, CALCULATIONS). 418-018:PAGE C-49 TEST ARTICLE, CONTROL ARTICLE AND VEHICLE PREPARATION PROCEDURE 5. Add a magnetics bar to hevessel, lace on amagneticsi plate and 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). `mix until uniform prior to and during dosage administration. Wattenby Approveby Clarification: Date: 21a aout: TCh okXeDyn wf 7 No __(Yes{See attached clarification form.) 418-018:PAGE C-50 ATTACHMENT 4 TISSUE COLLECTION AND PROCESSING 418-018:PAGE C-51 ATTACHMENT ProtocPoalg4e16t.t01t6 [E = T BToyooE3t1 Go|nPFooeksiUes Fouad Dood | + bie Tissue Collection and Processing : Coens | swepesre | anaes [Tesnn 5=20 Tah e LroOsLOL [Toe |Glam RT|| rPoolbmgitBe50 Err E S The-- mr e cccoonrl.F-- motr Te PDtame | PBoecmheis a a ne Cohamm cFBroorubmeigenst a edo Te wr PFroossse BT ane [79 Torte TinIgClyLE [MDiookd P[4omioenn oe 1 Tem VTistiumm TTEE) aa ioe ER Tabu T~ome a Torcomen| ciritohnpe Bienen cl mg a Bloat | Pooled [seVens TsTe Troe [am [Em T|Twantoawvoisnmoe ] T= ros || | T 5 e || [[ oTrV Fmemii Woese vanes s ]Fo i [= TSoteteenme]r w ER RT Room Tempers EN iron Mian nPBooilcohnciclhem non metn: 418-018:PAGE C-52 r2 FRIMEDICA Arg5u0s5 RSehseeaerhcoyhsDLhaibaeor..atBoPriAues1,o95InAn2.1 TelTeephtoan:e 5(1159))444433.85751807 e-- e ------------------ PROTOCOL 415.018 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 3 Storage (page 4of the protocol): "tTehmepesrtaotruargee.conditionsfothe Mevalonic Acid should be frozen, rather than room "This change was madea the request of the Sponsor. 2 Shipping Instmerions (page 6 ofthe protocol): The correct address for Dave Ehvesman is the following 3DaMveCoErbproersamtanToxicology 3StMPCaeunl,teMriBnunielsdoitnag 233561.404C-148 Telephone: (651) 733-5070 Telefax: (651) 737-4754 Any revisions made to this finalized amendment must be made by subsequent amendment. ReasonforChange: Thi change coecs the protocol. 418-018:PAGE C-53 PoFol sA1T80t18 Tod;t v& DeourlD.Doo ARTssspoewe 2 Resmi. YoLw. kPE. DPPv s T Ds ot n00 Associate Director of Research ASStoo Diver te Director of RY CrarrpatereLseeobno,nsViMAusDii.olnalleAntimalsCarecDate M SeaaymiM)oniLs toocro sMSe. d tDaeoo Ue Commitee Srl SELafhy Sige TAeormaBtuereSnthfytMPHoDn.oDABT. CH Date Am evisons made to hs inaized amendment must be made by subsequent amendment. PRIMEDICA 418-018:PAGE C-54 enobrebeanrt FOTO eae ONEEGESNETRAETIaORNLRCEHPARLOLDEUNCGTEIOANDSTNUODLY OIFNVPERSOTSI-GMAETIVOANLOINIRCATASCID SPONSORS STUDY NUMBER: T4085 n Alternate Study Monitor (page 1 ofthe protocol): The telephone number for John Butenhoff. Ph.D.. DABT. CIH is (651) 733-1962, rather than (651) 737-1962. SU -- 3 Sample Ants and Shinssions age 14oftheprove TT he ellent psa wilLpoLswotetpaorargrepsh iecrhssnitees tsht eirm and ver put pp ---- Serum samples cllcted fom ll dan sn pups wil arly for tol cholesterol, glucose, total and free Ts, Ts and TSH levels, LDL, HDL and a le oreeoto asartetToiFean To evr, The 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: Pro`toAcmoeln4d1m8e.n0:128 Page Dr. Saroj R. Das Ani Lytics, Inc. 200 Gerard Street, Suite 2000 Gaithersburg, Maryland 20877 Telephone: (301) 921-0168 Telefax: (301) 977-0433 Reason for Change `tTihsissuechsaanmgpelewsaasndmatdoepraitortihteizreeqtuheesutsoefotfhseaSmppolnessorforinaonradlyesretso. include additional 3. Scheduled Sacrifice - F1 Generation Pups (page 20 of the protocol): [pEefrfleicttteirv,esDaamtpel:es3fNroovmemmableera2n0d0f0e]maFloerpRueppsliccoamtebi1n.ebdl,ooradthsearmptlheasn pwoilollebde ppeorolsecxd. ASpapmrpolxeismwaitlellbye1amlLiqouoftebdlaosodfowlillolwsb,e rtartahnesrfetrhraendasindteosscerriubmedseipnapraartoargrtaupbehs3:. any srpeumaniinniancgenbtlroiofduwgiellanbed tthreanrsefseurlrteidnginstoerEuDmTAwi-lclobaetedidvitudbeeds.intTohtewsoaamlpilqueostsw.illThbee first aliquot (approximately 200 uL) will be used for PFOS analysis. The second galliuqcuooste.(atoptparloaxnimdaftreeleyTs3.0T0:uLa.ndofTsSeHrulme)vewlisllanbde LusDeLd.fHorDaLnalaynsdistroifgltyoctearlicdheolleesvtelesr.ol, tarcacnosrfdeirnregd10intthoeopnreioarliitqiueost.desRcersiubletdining isteermum1.waiblovbee. prAinoryitriezseudltaisnfgopllloawssm:a wcliilnlibcael cPhFeOmiSstsraimepslefsi.rst A(lalccsoarmdpilnegstowitlhlebperiiomrimteideisadteeslcyrifbreodzeinn oitnedmr2y,iacbeoavned) maanidntthaeinned frozen (<-70C) until shipment for analysis. Reason for Change: `This change samples for was made analyses. at the request of the Sponsor in orderto prioritize the use of 4. Scheduled Sacrifice - F1 Generation Pups (page 20 of the protocol): ([EwfofecltiitveersDpaetre:sa1m7plNeowvietmhbinerea2c0h00d]osaFgoer Rgreopulpi)c.ateSa2,mpblleosodwislalmbpeleasliwqiuloltbeed apsooled bfoelltorawnss,freartrheedritnhtoanEaDsTAde-sccoraitbeedd itnubpeasr.agTrhaephr3e.maAipnpirnogxbilmoaotdelwyil0l.b3emtLroanfsfbelroreodd iwnitlol sseerruumm swielplabreatdoirvtiudbeesd. inTthoetwsoamaplliequsotwsi.ll Rbeesuslptuinnginpalacsemntariwiflulgebeantdratnhseferrerseudltiinntgo one. aliquot. The serum will be prioritized as follows: clinical chemistries fist (according Any revisions to this finalized amendmentmustbe made by subsequent amendment, 41S-018:PAGE C-56 Pro"cAomlen4d1mo8ei.n0et1823 btoetfhreozpreinorointidersydeiscecrainbdedmaininittaeimne2,dafbroovzee)n a(<n-d7t0heCn)PuFntOilSsshaimppmleenst. fAolrlansaalmypsilse.s will ReaforsChoangne: "sTahmipslcehsanfgoreawnaaslymseasde a the request of the Sponsori order 0 prioritize he use of 5. Scheduled Sacrifice- FI Generation Pups (page 21 of the protocol): ((EbfyfsecetxivpeerDalittete:r).3 TNhoyvreomibdserwi2l00b0e]reTthayirnoeiddisnwnielutrbaelebxucfifseerdedfr1o0m%eafcohrmpaulpi.npfooorled possible future evaluation. Reason for Change This change was made at the request of the Sponsor. GH l; orgeL Deartod ve, Phi D. Ac BT s Dute 4x wd/ hd z2varvo Riyblond G. Yorks]. DABT Date Associate Director of Research AStsusdeyisDtiereDcitroerctof of Rbsearch Fhe tld zor: 7a C. Lebo, V.MD. Date assaf atin DeannJa. Lucker, MS. witDeat0e CahnadirUpseersCoonm,mIintsttieteutional Animal Care Study Monitor PE Pl 2 Bullet Sai froeo JAolhtnemBautteenShtoufdfy, MPohnDi.t.orDABT. CIH Date Any revisions to this finalized amendment must be made by subsequent amendment. 418-018:PAGE C-57 7,PriFRIMEDICA Amy5s05RSesearychDLiavbeo,raBtruicn, IgcA. TelephoHneo:rs2ha1m5,) P4A43189701404 "Telefax (215)443.6587 PROTOCOL 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. ControlArticles (page 3ofthe 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 (pag3eof 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 preparationof the mevalonic acid control and PFOS plus mevalonic acid formulations 3. Sample Analysis and Shipping Instructions (page 14ofthe protocol): PVelhaiscmlaemCeovnatlroonli(cGarcoiudple1)v,elMsewvialllobneicmeAacsiudrCeodntfroolth(eGfroolulpow2i),ng1.d6omsagg'ekggroPuFpsO:S + Mevalonic Acid (Group 3), 2 mg/kg PFOS + Mevalonic Acid (Group 4), 1.2 mek PthFaOnSall(Gdroosuapge11g)r,ou1p.s6.mMgekvgalPoFniOcSa(ciGdroluepvel1s2)maanydb2emmge/aksgurPeFdOiSn t(hGerloouwper13P),FrOaSther dosage groups after evaluation of the resuolftthse above analyses. Remaining samples will be retained at Primedica Worcester Laboratories or the Testing Facility uniil notification from the Sponsor for disposition. Any revisions made to this finalized amendment must be made by subsequent amendment. 418-018:PAGE C-58 Reason for Change: "This change was mada he ques ofthe Sponsor fF--aT--e 4. Sample Analysis and Shipping Instructions (page |of Amendment 2 to the protocol): Total and fee Ts, To and TSH levels wil be measured or he 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 efvraeeluTast,iToyn oafndthTSreHsulletvseolfs mthaeysbbeovmeeaasnaulryesde.in the lower PFOS dosage groups after Senntetiee Tinto wma isnt ie Sen Cora. E Deaove,t PLD Do ABT n Date. RimabdeG.dYolrk,y nD), DABT D0a0te.01 alee DreotfReostna ASSeoeaaed DiDreicrteocrtor searchand ssn lidioldts ena C. Lebo, V.M.D. Date hn Cons Chairperson, Institutional Animal Care hissed 11500 DeannJa. Luebker, M.S. Date Study Monitor Golan 2. Cozi 187 oA or JAolhtnemBautteeSnhuodfyf MPhoDn.oDABT. CIH Date beAcrp euistioyns sad ths Soatzat smanihsnt suas be malo by whveamsnt r2 FRIMEDICA 418-018: PAGE C-59 ArgSu1s5RSehseesahrcyh DLraiboer,atBouriielsd,iInA.g TelephoHnoer:s2h1am5,) P4A43-189701404 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 Amendment4 - 18 April 2001 I. El Generation Pups (page 10 of the protocol): [Effective Date: 21 August 2000] Day | of lactation (postpartum) is defined as the dayofbirth and is also the first day on which allofthe pups in a ltter are weighed. "The pups will not be individually weighed; pooled litter weights will be recorded. ReaforsChoangne: "This change was made to be consistent with Tests, Analyses and Measurements FI Generation, Body Weights 2. Scheduled Sacrifice- FI Generation Pups (page 20, 21, page | of ATTACHMENT 4 and item 5 ofAmendment2 to the protocol): Histopathology will be performed on the heart and thyroidof one male and one female F1 generation pup from each of four liters from the Vehicle Control group (Group 1) and 2 mg/kg PFOS group (Group 13). Ifany difference is found, histopathology will be performed on the heart and thyroid of one male and one female FI generation pup from eachoffour litters, when possible, from the 0.4 and 1.6 mg/kg PFOS dosage groups (Groups 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 Any revisions made to this finalized amendment must be made by subsequent amendment. ReafosrChoangne: This change was made at the request ofthe Sponsor 418.01PAGE C-60 --Nnen-- iments oz [leg coin 2Os. ``AGsikporigaete.DiDreeacrtloovreo,fRPehsDe,aDchABT Date AsuRasynnigohddSctiuadtyeDGiD.irreYecoctrtokor,r tooRB]odDeaArcBh T Date fran din ine Lipson dit tint C1hnadirUpseerCsHo.onmW,modIdirndts,teientiDo.naVl.AMn.imal CareDate DSteuadynnMoaJ.niLuteobkrr, M.S. Date loan. Gailonty peal ZF, 7 John Butenhoff, PhD. DAB, CIH Date Alternate Study Monitor aAnmyenedvmiesnotns made to this finalized amendment must be made by subsequent 505 Sheehy Drie, Bid.A HoTTreelskehfpoahcomn,e2P:1A(52)1514)94043444335867710 418-018:PAGE C-61 ~~ ARGUS RESEARCH Charles River Laboratories Discovery and Development Services PROTOCOL 418-018 `ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 `Amendment 5 - 4 February 2002 1. Shipping Instructions (page 6 of the protocol): [Effective Date: 29 January 2002] Homogeneity and concentration samples shipped to Dave Ehresman at 3M reshipped (frozen on dry ice) to: Corporate Toxicology for analysis will be Emily R. Decker (Principal Investigator) E30x5y8geRnesReeasrecahrcDhrive State College, Pennsylvania 16801 Telephone: (814) 272-1039 Telefax: Email: (814) 272-1019 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 OipnsepreacttiinognsPraoncdedauudrietstohfe trheastpefacctiilvitey.resSuultcshacnridtirceaplorpthsasaeccionrsdpiencgtitoontrheepoSrttasndaanrdd audit York. reports will be submitted by The dateofthe inspections that and facility to the Study Director, Raymond report submissions will be incorporated G. into a QAU statement generated report for protocol 418-018. by Exygen Research for inclusion in the final Any revisions to this finalized amendment must be made by subsequent amendment. Reason for Change: This change was made at the requestof the Sponsor. 418-018:PAGE C-62 Pc--a 1-- 5[.i0v1]8 - /L Glorge #. Dearlove, PhD, DABT Associate Director of Research Date ond G. York, |Ph.D., DABT Date AasnsdoSctiautdey DDiifrector f Research Moher lfAdirolindcbs ised Jit colosioon oh / Theresa H. Woodard, D.V.M. Date Chairperson, Institutional Animal Care and Use Committee Deanna J. Lueber, M.S. Study Monitor Date Db55 7 JAolhtnemButteenShtoufdfy, Monier Ph.D., DABT, CIH Date Any revisions to this finalized amendment must be made by subsequent amendment. 418-018:PAGE C-63 VanI. 15048 905SheehyDrive, BidgA. a (215) 443.8710 Teiefar: (215) 443-8567 y ; ~~ ARGUS RESEARCH Charles RiverLaboratories Discovery and Development Services PROTOCOL 418-018 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/ CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 `Amendment 6 - 9 October 2002 1 `Shipping Instructions (page 6 of the protocol and Amendment 5, item 1): w[iElflfebcetiavneaDlaytzee:d u1s4iFnebtrhueaLryEY2M00S2/)MSThceonsdaimtpiloenss fsohuinppdeidnthoeExaytgaecnheRdepsacgaersc.h Besson for Change: Tmheitshocdhoalnoggeywtaosbmeaudseead fotrhethreeqauneasltysoifsBoxfytghensaRmepsleeasr,ch to clarify the analytical 2. Shipping Instructions (page 6 of the protocol and Amendment 5, item 1): [Effective Date: 16 September 2002] The e-mail address for Emily R. Decker is emily.decker@exygen.com. Any revisions to this finalized amendment must be made by subsequent amendment. Reason for Change: 418-018:PAGE C-64 Po-- s iTbst `This change was made because the e-mail address was changed. 1 3) ben Sada)orado rk E. Dearlove, PhD, DABT Date `Raymfnd ZV G. York, fh.D, PABT Date jssociate Director of Research iAnsds Study DDiirreeccttoorr & search DRenea Cn. Lebo,CVALD 0Due MDelanana J.rLucboek, lMi5.ct soDaltuete Member, Istutiona! Animal Care Study Monitor and Use Committee Frm Balada softs. JAolhcnrmBautteenShtoufdfy, Montor Ph.D., DABT, CIH Date. Any revisions to this finalized amendment must be made by subsequent amendment. E en RESEARCH ProrogveRresRecsh. 415-018:PAGE C-65 LC/MS/MS System and Operating Conditions (Turbolonspray) Mass Spec: PE SCIEX API 3000 Biomolecular Mass Analyzer Interface: SHaCrIvEaXrdTiunrfbuoslioonn pSaprraapy Liquid Introduction Interface Computer: Dell UltraScan P1110 Software: PWiEnSdcoiwesxANnTalyst 1. HPLC: HewlettHPPacQkuaartdP(uHPm)pSeries 1100 HHPP AVuaocsuaummplDeegrasser HP Column Oven HPLC Column:Genesis Column Temp.: 35C Cs (Jones Chromatography), 2.1 mm x 50 mm, dp IMnojbeicltieonPhVaols.:e 10 uL (A): 2 mM Ammonium Acta in ASTM type | water MFolboiwlReatPeh:ase (B0):3 mMUentmhiann.ol Ti0me %0A 4B 70 0 00 1100 I5o 66 4 4 Iinsmtaruymebnet npeecrefsosramrayncet.o adjCuosltutmhnesHwPitLhCdigfrfaedrieenntt diinmeonrsdieorntso (oep.tgi,mi2z.1e mBemtasixl 3C0ig memt)c) acnadn cboeluumsneds, fprroomviddiefdfereeqnutivamlaennutfaccthurroemrasto(gKreaypshtyonies obtained Tons monitored: Anale Mode PFOS neguive Transition Monitored 499599 Xp RetenAtpipornoTxiimmaete(min 440 0 ESE Rao, un 3 as I en RESEARCH "Prin Koh 418-018:PAGE C-66 On the a day-to-day basis, the batch of mobile phase, retention etc. times may vary slightly depending on Example Tune File Parameters The following values are instrument to instrument. provided as Also, these an example. values may be Actual values changed from may time vary from to time in order to optimize for greatest sensitivity. `OTnhicsettuhneeifnilsetriusmtehnetniussteudneddu,rtihneg orpouttiimniezeadnaplaysriasm.etTerhseatruenisnagvepdroacseadu"rteunmeafyileb"e repeated as necessary to ensure optimal sensitivity. `Tune File Parameters Controls: IS-Tospray DP-Declustering Potential FP-Focusing Potential EP-Entrance Potential CE-Collision Energy CXP-Collision Cell Exit Potential DF-Deflector CEM-Channel Electron Multiplier set 42000 -5L.0 2300 17000 5 300.0 28000 Gas Flows Set Nebulizer Gas Curtain Gas 12 13 Collision Gas TIS Temperature 4 350C Calibration Procedures a Inject the same volume (between standard (prepared in MeOH) into 10 the to 20 uL) of LC/MS/MS. each calibration b. sUtsaendlairnedarc,ur1/vxesweairgehtgeednesrtaatneddardfocrurevaecsh fsoertqubayntilfiinceaatrionr.egrLeisnseiaorn fuaslilnigngthoeutaspipdreop+ri3a0t%e,sobfatsweadreosnysittsemc.alcuAlnatyedcacloinbcreanttiroantisotna,ndmaradys obfeceaxlcilbruadteidonfrstoamndthaerdcsaltihbartamtiaoyn cbuerveex.clHuodewdevmeurs,ttnhoetteoxtacleendum2b0e%r of the total number of standards injected The comelation coefficient (1) for calibration curves generated must be 20.9925 (20.985). If calibration resogfall outside these SEESBl on usa I;E5o20n22c1mo03m210 E e RESn EARCH "FrownRe 418-018:PAGE C-67 olpimeirtast,iotnh,enanadpptrhoeprriealteevansttespest sohfosualmdplbeestamkuesnt tboeardejaunsatlyiznesdt.rument Sample Analysis Inject the same of each sample, volume used fortification, for the control, calibration standards (between etc. into the LC/MS/MS. Each 10 to 20 uL) sample must be analyzed in duplicate. a. Sintcalnuddaerddsin caonrarneaslpyotnicdailngset.to at least six concentrations must be b. Inject inject an entire standards sientteorfspsetrasneddaradbsou(stixe)vaetryth5e-b1e0gisnanmpilnegs.of the All run and sample injections must be bracketed by standard injections. c. Tdehteercmoinnceedntfrraotmiontheofstaenadcahrdsacmuprlvee, fortification, based on the control, peak area etc. of is the manuasltytberianckaeltl srteasnpdoanrsdessionjfecttheed rdeusriidnuge afsoeutn.d Tihnetshetansdaamrpdleressepto.nseIsf tnheecesstsaanrdya,rddicluutrevethreansgea.mples and re-analyze to give a response within R360ER, Bean, us 1F6002i132r10 `eygencom APPENDIX D| DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 418-018: PAGE D-1 DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY I. Rats were ordered to weigh 175 & 10.200 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 0 go on study. 2. The following dosage administrations were performed from two minutes early to 60 minutes late, NDuomsbageer DoGsraogwpe_|_idenifcaiion AEadrmlyiniosrtLearteed| Rat Number VCeohnircolle ||1232SSEEPP000O DDSsH2 23 1TIS10r5--101115511144 114000CCTT0000|| DDSS474,3,DG0sG10-4 33 11009--91100099111144 214000CCTT000 Dsps391 23 11105-9101105114 2l4o0nCoTv0o0o|| DGs2G,1191-14 22 10109-9110039114 7 AcMiedvaCloonntircol ||1252 SSEEPP0000 DBSS? 2 ISIS, 11517 3 11515-11528 2180S0ECPT00O0| DS4D3S,10560 23 1110592165.-1101952288 114400CCTT0000|DS4p7,sD3G1s 1-4 33 1110591155. 1110592288 2l0oCNTo0v0e||D~Gs3,D96.2101-14 22 10911509-2170928 3 TPoRmOghS+e || 2126SSEEPPO0DD DSDT9O 2 10930 3 11529-11543 MevAadlodnic | 2[810S0ECPT0o00| DS4S31.5DGO 23 1101952299.1110594432 141.400CCTT0000| DDGss321-4 33 1110592299.-1110594432 240CT00 | DGs 12.14 2 109219019304932, "2+mMge/vkagloPnRiOcS|| 2126 SSEEPP0O0D DDSSTOO 37 1T1059443- 1110595475 Add | 12080SCEPT00O0| DS$S413,5DG0 23 111504-9-111045955376 114400CCTT0000|DS 4D7s.D3G1s0-4 33 1115044.9-111045955376 [7| oe |mNeOVmO| DDSG2105 2 TI1S0-95121571 DS iv uCsoncrdola_s|an abbr1ev0iNatoiovnOf0o|dayofsDudGy.13DUG used a a2n abbreviation1f0or96d7ay oF (presumed) gestation. DL is uscda a abbreviationfo day of lactation. 418-018: PAGE D-2 DGorsoaupge_| densification| _Date | TPoRmOgSk+s || 2281SSEEPPOO00 Cholesterol | 017100CCTT0000 0211N0oCvTo0o0 | 7 Zmeghkg PROS|0127NSoEPvOoOo| + Cholesterol | 12810SECPT0000| 120CT00| Day obfssSudy SS1254 DDSGS DDGG2210..DLL22 SD1SS5 DDSS4454.,DDGG00 NDuomsabgeer | AEdarmliyniosrteLarteed 22 22 22 22 22 2 200To0| DGs9. 11 2 0o1INNOovVo0o0| pGG261 22 02NOV00|DS 50.D7Gs4.5. 2 17N0V00 bLI 2 T1572.I-111S1558855 1110597824 1097120,98130973 1s8Ti0,98151585 11150869--119019549985 109910,099140992, 10981-0599110989, 1099171.59110998 11591-11599 11386 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. Atleast one event of dosage administration and/or postdosage observation was not recorded for the following rats. GDroowuipe [TT Weniification Rat Number VehicleConrol |28SEPO0| DSIS |11502-11512| These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available 10 evaluate this parameter. 4. Dosage administration was not performed for the following rats oGrwoew Weniiication AdmDionsaigseteNroetd DSityudor 2 mg/kg PFOS + Cholesterol | Cholesterol |28SEP00_| DS15| 11596 | [C00 7 mgkgPFOS TT PROS |0IOCTO0|DST8|11623 | `These deviations did not adversely affect the outcome or interpretation of the study because it was a single occurrence for each rat 5. Onl 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. 418018: PAGE D3 [oRatmNumeber|T onl) |s al) | IE AOF S| [seeTis] Cos TrTs] `These deviations did not adversely affect the outcome or interpretationofthe study because only four rats were affected and the additional volume was small (0.1 to 03 mL). 6. On 10October 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 mLkg. These deviations did not adversely affect the outcome or interpretation of the study because the total correct dosage was administered 10 all of the rats 7. On 20 November 2000, DL 3, ra 11589 in the 2 mg/kg PFOS + Cholesterol dosage `group (Group 7) received 1.9 mL of test artile 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 asingle event. 8. Clinical observations, body weights and dosage administration were not recorded for the following rats [Gor| n wotonn [owe [ ooo T6 make PFOS NOVHD DDsG6159 DDGG2119 780v oD GD1G2 GD1GO2 GG1o0 Tmgkg PROS TTROVHD GD61122 DDGG 1110 DDGG 1110 DDGG iiI0 | sume| 1T1i04o1 1111002423 1H1e64s7 1111664590 H11e6s3t2 T1ie6sst3 1H1e6s3ss 1H6ess?s 111166690 These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate these parameters. 418-018: PAGE D4 [==] 9. Data for the following rats were not collected due to a computer filing error. [VehicleComrol |20SEPO0| DST |TIS01-1512| [5T Cholesterol Control 200C0T0 `These deviations did not adversely affect the outcome or interpretationofthe study because sufficient data were available for evaluation of this parameter. 10. iThe feed left and/or feed fed values were not recorded for the following rats. `These deviations did not adversely affect the outcome or interpretationofthe study `because sufficient data were available to evaluate this parameter. 11. Maternal behavior was not recorded for the following rats. [TT VehicleConvol |04NOVOO| DLT | 10914 | [2 Mievalonic AcidConwol |04NOVO0 | DLT | 10923. 10924 1.6 mg/kg PFOS + Mevalonic| 03 NOV 00 DLT 10977 "These deviations did not adversely affect the outcome or interpretation of the study 418018: PAGE D'S 12. The terminal blood collection data (time collected, volume collected, performed by and date) was not recorded for the following rats. [r [www| E wer |] Graogn Jenitcaion id DayofStody|-- Rat Nomi [ [8 61 |oTmrymkeePRnOacSgi+d Cphraos| se| 0m3aNNOoVveOr0|| bDGiasi || rToaomr || C0 imgiePros |2aNoveo | bis | tien | "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 forrat 10932 in the 1.6 mg/kg PFOS + Mevalonic Acid dosage group (Group 3) was not recorded. This deviation ddaitdanwoteraedvaevrasiellayblaeffteocetvtahleuaotuetcthoimsepoarraimnetteerrpretationofthe study because sufficient 14. The pooled liter weights were not recorded fo the pups of the following dams. [5Gro | CholMewneintcmaiiCononl |BNDOuVcOO| Li RattiNeummer| C8 oimsgrros--[z2Nove | bis | se `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. bTheicsaudseevisautfifoincideintd dnaottaawdevreresaevlayilafafbelcetttoheevoaultuactoemtehiosrpianrtaermperteetration of the study 418-018: PAGE D6 16. The weight of the pooled liver samples was not recorded for the male pupsof the following dams. Group leniiicarion [77VehicleConrol Mevalonic Acid Due DayofSuady | Rat Number |2SNOV0| DLS |mses | 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. 4 od so Lebar RAasys!ociaottehDGi.reYcotrokr,ofPhR.eDs.e,aDrcAhBT Date and Study Director APPENDIX E CERTIFICATE OF ANALYSIS 418.018PAGE E-1 Certificate Of Analysis FC-95, Lot 217 Mar 9,c 20h 00 RichM.aParyfder o`fThtihsessaetmpelseswsahsoawtnhaeslaympuzlseeitndogcLonCt/aMiSn,th"eHf-olNlMoRw,inUgFw-eiNgMhRt,peenrdceelnetmceonmtpaolsiatniaolny:sestechniques.The results ----Cdsox-- omy LL SoRSOT | mn Acdodnittaiionntahlleyf,otlhloewiisnogmmeordliesptreirbcuetinotncoomfptohseitsiaomnp:lewasdeterminedusing VE-NMRtechniquesandfoundto CFIC S07 05% |oom ey| | CF3(CF2)-CF(CF-){CF3),- 80; K Ce wherex+CyF ismai3CnF-lCFyy)x5- ,anSd0x7= 0,y#0) 5 ||l cp omopmtvir| ei" za| s Lembo on,Te pha Cb F3),r C-w{a ChFear)n exSi0c s5mKsh inl. y6) 07 ey| | CF3-(CF2) C(CF>)7-(CF2),- 30, K ismai 4, anndl x #y 0) 418-018:PAGE E-2 . SIGMA| CAi iRrrEeiEmeim DE iMEALNiD TEeAcED CTEacRTTonIEFICATE roma: OF ANAL cit 05 YSI S ors vor 74-3600 i FT e aein eo sA R Ocsrorc e mm creepsz r Sev: SELOp vee Coons _n h _TwR mE BERR pRvE W SeWOEErewe _T_o_ EE wPT wEmL EewET sacT s ee TooEm 2T % wm mee TITRATION Pes 2.2% QC ACCEPTANCE DATE JULY 2000 BD SUeBolEoRZnR mon--ssnvicss H Ss A1E9e1G7e t R2,0E000S, sa781Rtr/tYXSB1,ee It13EeEsocsn011 1 mrinn ctAs i1 1s a LS pe SE tpoosns A APPENDIX F HOMOGENEITY AND CONCENTRATION ANALYSIS REPORT 418-018:PAGE F-1 STUDY TITLE One Generation Reproduction Study of PFOS Mevalonic Acid/Cholesterol Challenge ind 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: 14-272-1039 STUDY SPONSOR 3M Corporate Toxicology St. Paul, MN 5514-100 PROJECT Study Protocol Number: 418-018 Exygen Study Number: 023-069 Total Pages: 21 418-018:PAGE F-2 Exygen Study Nos 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 Rescarch. fPriyncipka.lDInevesrtigator Exygen Rescarch \ ARragyuasnRedseGa.rYchorkgh). DABT Lea wna bt?) SDpeoannsonrJa.ReLpureesbeknetrative(M.S. 3M Corporate Toxicology DateJofsofe Hwovor Due L067200 Date Exygen Research Page 2021 418-018:PAGE F-3 QUALITY ASSURANCE STATEMENT Coens nd entitled, "One Challenge and NOEL IvcsigiinRoans ll iene pesvee pest for Generation Reproduction Study of PFOS -- Mevalonic Acid/Cholesterol NOEL Investigation in Rats." All reviewed phases were inspected for Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Study Director and to management. backports Drei Phase Inspected Management SponsorManagement go, Coton wm se oem I L Pome Angelg Morgan Quality Assurance Auditor A Date Exygen Research Page3of 21 418-018:PAGE F-4 Exygen Study Nos 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: E30x5y8geRnesReeasrecahrcDhrive. State College, PA 16801 (814) 272-1039 Principal Investigator, Exygen: IR. Dfcker D SEcxiyegnteinstResearch J Duc AY Exygen Research Facility Management: Richard A. rack,DE2D. PErxeysgiedenntResearch Study Director, Argus Research: re A RAarjgus jRdesG.earYcohrk, PHD.,[DABT hFue~our-or po. Date `Sponsor Representative, 3M: Ltonarblen Deanna J. Luebket/ M.S. 3M Corporate Toxicology bor Date Exygen Research Paged of 21 418-018:PAGE F-5 Exygen Study No.: 023.069 STUDY IDENTIFICATION One Generation Reproduction Study of PFOS --Mevalonic Acid/Cholesterol and NOEL Investigation in Rats Challenge STUDY PROTOCOL NUMBER: 418.018 EXYGEN STUDY NUMBER: 023.069 TYPE OF STUDY: Analytical SAMPLE MATRIX: Dosing Solutions TEST SUBSTANCE: SPONSOR: STUDY DIRECTOR: Perfluorooctanesulfonate (PFOS) 3M Corporate Toxicology St. Paul, MN 5144-100 Raymond G. York, Ph.D., DABT Argus Research 905 Sheehy Drive, Building A Horsham, PA. 19044-1297 SPONSOR REPRESENTATIVE: TESTING FACILITY: ANALYTICAL PHASE TIMETABLE: DeannaJ. Lucbker, M.S. 3M Corporate Toxicology 3M Center, Building 220-26-02 St. Paul, MN 55144-1000 Exygen Research 3058 Rescarch Drive State College, PA 16801 Study Initiation Date: 0821/00 Analytical Start Date: 0214/02 Analytical Termination Date: 0211502 Analytical Report Completion Date: xx/xx/xx Exygen Research Page Sof21 418-018:PAGE F-6 Exygen Study No.: 023.069 PROJECT PERSONNEL RTehseearScthu.dy TDhierefcotlolrowfionrgtphiesrspornonjeelctfrwoams ExRyagyemnonRdeseGa.rcYhorwke,rePahs.sDo.c,iaDteAdBwTithat vaArrigouuss phases of the study: Name Emily Decker Karen Risha Rickey Keller Tite Scientist/Principal Investigator Scientist/Principal Investigator Sample Custodian Exygen Research Page 60f21 418-018:PAGE F-7 Exygen Study No.: 023.069 TAOFB CONL TENE TS TILE EAE rorrrrrorssmmmsoms----------------Page GQUOAOLDITLYABAOSRSAUTROARNYCEPRSTAACTTEIMCEENTC.O.M.P..LIANCE STsATeEMEsNTmmm--30.2 CSETRUTDIYFIIDCEANTTIIOFNICOAFTAIOUNT.H.E.NTICITYs.e.si.ssmm----o--o--eoememsesesnnnenrenl PROJECT PERSONNEL.................. TABLE OF CONTENTS............. r-------------- r------------ LL ISTI OOFFTS FAIBGLUERT SE.Sd................... em-- s---------- r----------y L1I0STSOUFMAMPAPRENYDI..C..E..S......c...c..rnrnnnnA ensonssososnsososesose nn 8 20 3.0 OBJECTIVE INTRODUCTION BE erent rss nd rsesesesoios 45.00TREESFTERSYESNTCEEMM.ATERIALr---------- nm-- " ---- 8 6.0 DESCRIPTION OF ANALYTICAL 6.1 Extraction PIOCCAUIE. METHOD... coco reamsss-- 10 6.2 6.3 Preparation of Standards and Fortification CHIOMAOZEIPhY crore SOMOS ............. 10 smnsss-- 66..45 DIenssctrriupmtenitoSneonfsiItnisvtirtuyme. nt and OJ perating Conditions.a.. nmm------_kun 7.60.6EQXuPanEtRitIaMtiEoNn TanAdLEDxEaSmIpGlNe..C.alculation.......BE...... een 12 A 89.00RCEOSUNLCTLSU.SIONS... A im----o-- r -- 18 10.0 RETENOTFIDAOTNA AND SAMPLES........ A Exygen Research Page of 21 418-018:PAGE F-8 Exygen Sudy Nos 023.069 LIST OF TABLES Tablel. Summary of PEOS in Dosing Solution Sumpls............ ctPa?ge Figure 1. Figure 2. Figure 3. LIST OF FIGURES Page Typical Calibration Curve for PFO Chromatogram Representing a0. S..........c....... ng/mL. Calibration SR Standard for P FO 19 S...20 Chromatogram Represening Dosing Solution Sample (Exygen ID: 0200285, Set: 0214024). sanssiiss ---------- nein 21 LAISPTOPFENDICES Page Appendix A Study Protocol 418-01 (Exygen Study No. 023-069) and Amendments. 2 Exygen Research Page 8of21 418-018:PAGE F-9 Exygen Study No: 023.069 1.0 SUMMARY EpexryfgleunorooRcetsacnaerscuhlfonaantaely(zPeFdOS)doascicnogrdisnogluttoipornotoscaomlp4l1e8s-0f1o8r(AtphpeenddietxerA)m.ination of Recoveries for 0 the assigned PFOS in the dosing concentration. solutions ranged from 83% to 105% when compared 2.0 OBJECTIVE `The objective of dosing solution this study samples was to using determine levels the analytical of perfluorooctanesulfonatc conditions described in (PFOS), Protocol Amendme#n6t. 3.0 INTRODUCTION "sTohliustiroenposratmdpelteasilsustihnegrtehseulatnsaloyftitchaelacnoanldyistiisofnosrdtehsecrdiebteedrmiinnPartoitooncoolf PAmFeOnSdimnetnhte #d6o.sing Tnhuembsetrud4y18w-a0s18i.nitTiahteedaonanlyAtiucgaulststa2r1t,d2at0e00w,aswhFeenbrtuhaerystu14d,y 2d0i0re2c,toarndsitghneedanparloyttoiccoall termination date was February 15, 2002. 4.0 TESTSYSTEM Twenty-seven dosing solution Research on January 31, 2002 samples were received and logged in by Exygen frozen on personnel dry ice at Exygen and placed in frozen storage. Sample log-in and chain of associated with this study. custody Storage information records will can be be kept found in the raw data package 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 Environmental Technology and Safety Services. Exygen on May 17, 2000 from 3M Exygen Research Page 90121 418-018:PAGE F-10 Exygen Study No.: 023.069 The available matcrial PFOS information for the was stored frozen. reference material is listed below. The reference Compound ExygenControlNo, PFOS SP248 TCRNo, Purity(%) Expiration Date SD00S NA 01/01/10 "Themolecularstructure of the reference material i given below. PFOS Chemical Name = Molecularweight = Perfluorooctanesulfonate 499 (CiFySO5) a]I --o Ndeortiev:ed,`Tihsepnoetuatrsasliummolpeecrufllueoaronodcsttanaensdualrfdofnoatrem [fCr4oFmi;wShOi5cKh],PFmOolSec(ualnaiornw)eiisght 538. 6.0 DESCRIPTION OF ANALYTICAL METHOD 6.1 Extraction Procedure Each sample was diluted in methanol prior to analysis by LC/MS/MS electrospray. 6.2 Preparation of Standards and Fortification Solutions Ssttaannddaarrdd ssoolluuttiioonnswaosf pPrFeOpaSrewdearteapcroenpcaernetdraotnioJnanoufa1r0y01p0,g/20m0L2.by dAinssionldviivnigdu1a0lmsgtocokf the LO standard (corrected pg/mL fortification for purity and/or standard solution salt content) in methanol. was prepared by taking 0.1 From this solution, a mL of the stock and bringing the volume up to 10 mL with methanol. Afort0i.f1icpatgi/onmLstfaonrdtairfidcaatniodn bsrtianngdianrgd twhaesvporleupmaereudpby(0ta1k0inmgL1.w0itmhLmeotfhatnhoel.1.0 Apg/0m.0L1. pg/mL 10 mL standard was prepared with methanol by taking 0.1 mL of the 0.1 pg/mL standard and bringing to Exygen Research Page 100f21 418-018:PAGE F-11 Exygen Study No. 023.069 A set of standards containing PFOS calibration solutions in the following was prepared manner: by dilution of the 0.1 pg/mL various Initial Con. (g/mL) or ol 01 0.005 0.002 0.001 0.001 Volume (mL) 05 02 ol 10 10 10 0s Diluted to (mL) 10 10 10 10 10 10 10 Final Conc. (g/mL) 00..000052 0.001 00..00000025 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. 63 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 concentrationof0.0001 pg/mL. of PFOS. 6.5 Description of Instrument and Operating Conditions Instrument: Micromass Quattro Ullima Computer: Dell UltraScan P1110 Software: COMPAQ Professional Workstation AP200 Windows NT, version 4 HPLC. Hewlett Packard (HP) Series 1100 HP Quat Pump. HP Vacuum Degasser HP Autosampler HP Column Oven Exygen Research Page 11 of21 418-018:PAGE F-12 Exygen Study No: 023-069 HPLC Column:Genesis Cs (Jones Chromatography), 2.1 mm x 50 mm, 4p Column Temp. 35C Injection Vol.: 10 uL 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 CT) 10 0100 70 0100 75 60 40 no 60 40 Tons monitored: Analyte Approximate Mode Transition Monitored Retention Time (min) PFOS. negative 499599 44 Tune File Parameters CIoSn-Ttorsoprlasy 42S0et00 DP-Declustering Potential 510 FP-Focusing Potential 2300 EP-Entrance Potential 100 CE-Collision Energy 10 CXP-Collision Cell Exit Potential 5 DF-Deflector 3000 CEM-Channel Electron Multiplier 28000 GNeabuslFizleorwGsas S1e2t Curtain Gas 1B Collision Gas 4 TIS Temperature 350C 6.6 Quantitation and Example Calculation `TTwheenpteyakmiacrreoaliwtaesrsmoefassuarmepdleanodr cthaelisbtraantidoanrdsctuanrdvaerwdawsergeeneirnjaetcetded(uisnitnogt1h/exLfiCt/wMeSig/hMtSe.d linear regression) by the Micromass software using six concentrations of standards. The concentration was determined from the equations below. Exygen Research Page 12021 418-018:PAGE F-13 Exygen Study No.: 023.069 Ethqeuasttiaonnda1rdcaclucruvleate(dlintehaerarmeogurnestsioofnapnaalryatmeetfeorusn)dg(einnernagt/emdL,bybatsheedMoincrpoemaaksasraso)ftuwsairneg program. equations Then Equation 2 calculated for serum are shown). the amount of analyte found in pg/mL (the Equatio1n: Equation 2: Analyte found (ng/mL) = (Peakarea - intercept) slope. Analyte found (g/mL) = (anal. found (ng/mLx) DF) x Lug Where: 1000 ng DF = Dilution Factor Equation 3 calculated the percent recovery. Equation 3: Recovery (%) = Canal. found (ug/ml) x 100 sample conc. (ug/ml) An example ofa calculation using an actual sample follows: Dosing solution sample Exygen ID 0200283 (Set 0214024), where: peak area intercept slope. dilution factor sample conc. = 1380 = a3sis = 2m = 200000 = 400uymL From equation 1: Analyte found (ng/ml) = [1380413545 720773 = 18SngmL Exygen Research Page 130f21 418-018:PAGE F-14 Exygen Study No.: 023-069 From equation 2: Analyte found (ug/ml) = (LSS ng/mxL 200,000) x Lug 1000 ng = 30 ugmL From equation 3: % Recovery = 370 ug/ml x 100 400 pg/mL = 93% sNloitgeh:tlTyhdiisffeexraenmtplferocmaltchuelavtailouneswasshodwonneinustihengRrAouWndDeAdTnAu.mbers, and therefore may be: 7.0 EXPERIMENTAL DESIGN `The and set of samples throughout the cruonnsainstdetdhoef2a7nsianmpsltersu.ment blank, a set of calibrations at the beginning 8.0 RESULTS `ITnhdeivaivdeuraalgreecpoevrecreinetsrefocrovPerFyOS+ sitnanthdearddodseivnigatsioolnutfioornPsFaOmpSleins tahreeddoestianiglesdoliuntTioansblwaIes.. 93% 7%. A typical are given calibration in Figures curve 1-3. and representative chromatograms of a standard and sample 9.0 CONCLUSIONS `The concentration and results of this report. homogeneity of the dosing solutions were demonstrated by the 10.0 RETENTION OF DATA AND SAMPLES `wiWlhlenbethsehifpipnaeldrteopotrhteisspcoonmspolre.teT,hailsl dooriegsinnaoltpianpcelruddeatfaacgielnteyr-astpeecdifbiyc ErxaywgedantaRessuecahrcahs instrument logs. Exact copies of all raw data, as well as a signed copy of the final Exygen Research Page 140121 418-018:PAGE F-15 Exygen Study No: 023-069 aRneaslcyatriccahlarrecphoirvtesanfdoraltlheorpiegirniaold foafcitliimtye-ssppeecciifficierdaiwn d2a1taC,FwiRllPabretr5e8t.ainReedtaiinntehdesEaxmypgleens of reference substances are archived by the sponsor. Exygen Research Page 15of21 418-018:PAGE F-16 Exygen Study No. 023-069 Exygen Research Page 160f 21 418-018:PAGEF-17 Exygen Study No: 023.069 Table. Summary of PFOS in Dosing Solution Samples Exyeen Sponsor TTTPek eFovuned AFomwed SCao.w Rec is 0200283 Tor T 04 myn) BAUGD0 DE 200000 Aves nein) 1380 Lss (ppm) apm) 1 400 (0 93 C2000228545 3S0offG6MBOO44mmYgmmLl)2)BBAAUUGGO0) 22000000000 11555212 220095 1410s d4o0o l1o0s3 C008 000287 1101o20f082m0ynmly) SImSElSPE)POO 20 ND ND S000 995 132 ND 661 80 8 0200288 0002 10f2(0.16mynl) 10f2020 mpm) ISSEPOO IBSEPO 2S00000000 2156 704 02993% 146 is 21600 9 9 0200290 0200291 10f2(024mynl) 1012032 mynl) ISSEPOO ISSEPO0 200000 200000 1S06779 LLsu 2289 22800 9950 0200292 20093 1100f(62T(O0440mmpymnDl2)7ISSESEPPOOD0 220000000000 1L3S8S5 210506 3423 4400 19031 020094 020095 3of6MO4mynl)2SEPOO Sof6BOAmgML27SEPOD 0000 20000 123 113 191 176 3 32 40 40 96 8 000% 1or2OmymlOLNOVO) 2 91 No No. - 0200297 20095 110200162mmppm0 mOOLL0NNOOVV8OO0O S000 S000 21004026 21738 665 139 1$600 8 8 02009 0200300 10(2020mgmL)OLNOVOO 10(2(024mgm)OLNOVOO 00000 20000 743 863 0972 Lis 19% 28 20 20 97 95 C0001 C0002 1100f2200420mmggmmL))OOLLNNOOVVOOOO 2000000000 1103528 117490 23881 2400 88 C0003 1o20mpml)ISNOVO) 2 95 NQ MQ - 0200304 0200305 110o7f220@0I8Gmmiynndl)) ISNOVOO ISNOVOO S000 S000 21008012 133 28 665 141 1$600 83 88 02000330076 110o7f22(@02240mmpmll)) IISSNNOOVVOOOO 220000000000 SR0226 110059 221117 2200 105 ol 0200308 C0030 1100f220(30420mmglnl)) IISSNNOOVVOOOO 220000000000 1L144851 115939 33096 4200 19060 ND = Not Detected INQ = 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 Page 170f21 413-018:PAGE F-18 Exygen Study No.: 023.069 Exygen Research Page 13 0f21 418-018:PAGE F-19 ExygenStudyNo: 023-069 Figure 1. Typical Calibration Curve for PFOS a``boCraoafmsipicoonuntn1doufnnDea:moet:7s2F1oO7nS7:3 x099491.1300245 (RGeusnponyspoetyLpien:eaEr,xoOrnragl SiE,xcAurseea, Weighting: 1xAvsrans: ono 3700. x - Wo ws te 1s 20d ab Ws do as weoTM Exygen Research Page 19.0121 418-018:PAGE F-20 Exyeen Study No. 023.069 Figure 2. Chromatogram Representing a 0.5 ng/mL Calibration Standard for PFOS Corin a5vom snirs ra rm . Ec TTRRa Exygen Research Page 20 of 21 418-018:PAGEF-21 Exygen Study No. 023.069 Figure3. Chromatogram Representing Dosing Solution Sample (Exygen ID: 0200283, Set: 0214024) rp-- roar|n smom sn Ts sebns svn = wnEAGEwE= | htt | TT wh TRB IEETR E Exygen Research Page 21 of 21 APPENDIX G TEMEPRATURE AND RELATIVE HUMIDITY REPORTS 418-018:PAGE G-1 ARGUS [rr TemperatureLoacnadtiRoenl:atRioveomHu1m7idity Report Protocol Number: 418018 `Range of Dates: 22-Aug-2000 14:44 to 12-Nov-2000 08:59 | TSAapregceite-- sR:arnagt e: TToottaall NNuumoombffHDboaeutrarseP:oinrts: TBemlpoera7t9ure 196wn20 1955 | Rela"ti3ve10H0u0m%i%dity 195620 1955 Mean (25D): | MMMeaidxniiiammnuu:mm:: NNNuummibbeerrrooofffmPPPoooiiibnnntttssseLiiongrRwhangiel(:0 (%): | ms ea | ses am 5n780e 7570780 192v54 (6@000%)9) | 180T55 (60199)) Report Generated: 30-Nov-2000 at 1402 comments: REVIEWED BY:7 _ ) = oate: 3 2570 ARGUS 418-018:PAGE G-2 TemperLaotcuarteioDne:vRiaotoiomns17Report Protocol Number: 418018 "RangeofDates: 22-Aug-2000 14:44 to 12-Nov-200008:59 Temperature Target Range: Species: rat 0o7DrSSaeetpp3eo0o000 T0oi6r0mo0e Te66m33p8s.1t 64F 10 70F Date Time Temp. H=Voutaofrla"nTgeuermp.Hsi=ghTe_mpeLratu=re VFcutaof rlangueLpow Report Generated: 30-Nov-2000 at 14:10 + hese deviations cid not adversely afec the outcome or interpretation ofthe sic. "The folowing deviaton(s)impaced on the outcomeof he study as described: StudyDirector X re 7 Kore Date: 30- Mh#2-d) ARGUS 418-018:PAGE G-3 Relative HLoucmaitdiiotny:DReovioamti1o7ns Report Protocol Number: 418018 Range of Dates:22-Aug-200014:44 to 12-Nov-2000 08:59 HSpuemciideist:y rTaatrget Range: 0.0Dantze00 T2i20m0e RTH0.8H 30% to 70% Date Time RH. /ent rt vt tc ior env H=VouatoflRanSugSesRivHighe_HLu=mViad Goluytuofeange-Low Report Generated: 30-Nov-2000 at 14:18 he folowing deviaton(s) impacted on the outcome ofthe sudy as described: StudyDirector: / or {bx i Date: 30" MRO! ARGUS 418-018:PAGE G-4 Temperature and Relative Humidity Report Location: Room 18 Protocol Number: 418-018 Rangeof Dates: 05-Sep-2000 14:00 to 03-Dec-2000 13:59 ||| nTargeet Range: | im TToottaall NNumubemroobffHDoeauyrsrs:: | Moan (25D): | | MMMeaidxniiaimnmu:am:: | Numobf Peoinrts in Rango (%): NNuimomob ffPpoe bionir tsseLHoirgwhS.Goa) Report Generated: 27-Mar-2001 at 16:18 comments: TeSRT i2395075 | aro Relative Humidi | 1%21595075 w2 wos | se an Jomes | 82n332 29 oo| 2 (000) 0LI 3 I1 (0.0) RevieweD sv: J om 3260) 418-018PAGE G-5 ARGUS Relative LHoucmaitdiiotny:DReovioamti1o8ns Report Protocol Number: 418.018 Range of Dates: 05-Sep-2000 08:00 to 03-Dec-2000 13:59 HSpuemciideist:y rTaatrget Range: 0s0Daatze00 TOirmoe R0H2.H 30% t0 70% Date Time RH. H= Value out ofRrEanSge RHeilgihv_e HLu=mVialdueoGuytofrango -Low Report Generated: 13.Dec-2001 at 14:57 Tessdonors tarsscmor rotonsty. The folowing deviaton(s) impacted on th aufcome of the study as described Study Orca:ee = owe bcos APPENDIX H BIOANALYTICAL REPORTS (ANILYTICS) 418-018:PAHG-E1 Clients NEUE EERE CRIES, He ate SoLteeels INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 AJectecressionspec. Ell GpSexAgeCHOLEST GLUCOSE LDL-DaIRlHDaL-kDIR TRIG 418-018:PAGE H-2 Cite: ETT ER GIS, TO. Sat SEALER INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 Jjezeoseicon ession Spec. | GpSexAge GTHOLeEST GfLUeCOSE LgDaLp-eDIR HgD rL- ToRIDnG IR ence J 96 319 0 0 104 418.018:PAGE H-3 INCORPORATED 301-921-0168 Fax: 301.977.0433 Client: AS0R8GUSSHEREEHSYEARDCRHIVELABORATORIES, INC. DDaattee CRoelcleeicvteedd:: 12/08/2000 BHOURISLHDAIMN,G APA 19044- Date Reported: 01/16/2001 (318) 44378710 Client No. 1028 Study: 418018 DAMS GD21 Species: RAT NAocwcessionsopt ec. BBB 00o00o33j00oi1i6s3;l 111000999333310 BBB000000333000iiieeesie 11100099833353 BB 00003300116687 1100984307 MSe8a7n Group 3 GpSexAge 333 EEEFFF 333 FEEEF 33 FEF GMHGO/LDELS|T GMLGU/CDOLSE IDMGL/-DDLIR 13771%5 i11i0ss2 31110 1128705 11399773 1isi 8740 10887 25 M51a B 1gs,U1 S11.8 FMDGL/-DDLIR TRMIG/GDL i24s6 i3252s81 23333 52272736 335 323531 #351.1 830k5.3 BBB o00o00333o00i11767d90 111000999444765 44& FFFEEE BBB 000000333000111777332 11100008985415s 444 FFFEEE BB 00003300117785 1100888834 44 FFE eS7a5n7 Group 4 1357444 1258886 18s0 i5usn6 118381773 8887 11i00g28 Fr1)3 82510 11EtH3H] 18050 110272 brFS} 2is 324238 838.a9 1ad06 95 6 11728 32a730.8s Reference Range 856- 317sa- 0 - 08 - 3fo-s Page 3 + JIBOISPAGE ESToRATED SSH Se 20Sees (215) 443-8710 JAesecmalcenession spec. Gp SexAge Ell CHOLEST GLUCOSE LaDLl-DIR a HDL-DIR TRIG Mean 79.3 98.5 3.9 a1 191.5 ree aT Se a INCORPORATED 301-921-0168tt BeFax: 301-977-0433 (215) 443-8710 Accession spec. B 0030193 10985 Gp Sex Age CHOLEST GLUCOSE LDL-DIR HDL-DIR TRIG 7F 89 95 16 61 111 418-018:PAHG-E6 INCORPORATED 30S 8 EERY DRIVE (215) 443-8710 301-921-0168 Fax: 301-977-0433 Bate Rezefvest" 12/08/2000 JjicecesaionSpec. | GpSexAge TT4TaOTAL1p3 LTOTAL FaRpEEn15 TwSHy| , FfREeE T4 418-018:PAHG-E7 Client: agus RIESNECAORCRHPOLRABAOTREATDORIES30,1-9T2e1-.0168DateFaxG:oi30s1e-m97t7a-n0:433 (015) 443 8710 AJotecasicon ession spec: Gp Sex Age TT4,RTOTAL T3,TORTALL FEREEE T3 TTSHEFRE T4E 418-018:PAHG-E8 Client: gLg T EISNACEA ORRPOIRNATAEDRI,301-9T2r1e-.0168Bpa2t8sFaxSS:Ro3Lr0Ls1T-t6Eo7Ec7-Ti0n433sasnnaone i se seis Te ve (215) 443-8710 NAecscession S3pBac. | GpPSSeexxAAggee TT4H,,TT0OTTAALLTT3,ETTOTAL FBREPEET3 TRSHoa FRRAEUEELT4 ABOISPA1GE Citents pucus KIENsCENORREPOLRABAOTREATDORIES30,1-9H21e-.0168BatsFaxS:ei30l1s-c97e7a-d04:33 (215) 443-8710 Fs Accession spec. Gp Sex Age TT4,HTOTALTG3,TLOTALFGREEET3 TSH FREETa Mean 013 52.031 .235 1,452 134 418.018PAGE 10 Client: A18R58G,USSERISESNEECAORCRHPOLRABAOTREATDORIESS,oroIeNrCo.resBDaatteeFaSCxoRlsRloSvgEacRrthYeo"dm:ss1208/2000 BBSeREERRBEiCEEBh sooue- Date Reported: 01/16/2001 Cutent wo(.2151)028443-8710 Study: 418018 DANS GD21 Species: RAT aReesanaionS8pec. p5Bpooogomaaomeimsamseeii1st3lees: BBBBooOooniSameSsaasItrs yeeagn Group 7 Te Sex nae HBaBo R13AEN0T BBRTRR aNh.a RETEBA 777 EF ii eea8:eo8888d3a31Eya SeoalBaef IzbnaEfsl oo0o.llt3f3 7777 OF 8geg:::58e8888 3hBine%id 8gv8i5en ES sae 8188:8B3]8 5 2T5o aa0e aNaWn Reterence Range 33. 8so. 98 - S srohiase ApprovedSag-e---T=momo-ronn1M0illoeen Date ---ilefdl 418-018:PAGE H-11 INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 AcJescteer ssion Spec. opsex Age OR CHOLEST GLUCOSE i LDL-DIR a HDL-DIR TaRI,G 418-018:PAGE H-12 CAXTICS consi st 1 cnt vo zm INCORPORATED 3019210168 Fax. 301.977-0433 Client: A5R0GSUSSHEREEHSYEARDCRHIVELABORATORIES, INC. DDaattee CReoclelievcetde:d: 12/12/2000 BHOURISLHDAIMN,G APA 19044- Date Reported: 04/17/2001 (215) 443-8710 Client No. 1028 Study: 418018 DAMS LDS Species: RAT ANoc.cession s3pec. BBB 000000333000888099088 111111000332219 BBB 000000333000866000133 111111000333135 BBB 000000333000666000546 111111000333887 MSe5a7n Group 11 Gp sex Age CMHGO/LDELS=T GMLGU/CDOLS~E_LMDGL/-DDLIR HMDGL/-DDLIR TRMGI/GDL 1i111 EEEF 55&80 111578828 i21 ia331s 2f4i73s0 1oo1 FF 84i4 11778983 884 i3317 i1s3e2 1I11I1 FEFF 53E80] i53s30o 3 313875 31735880 851431 i1s89.4 11.%8 1307:773 51882% BBB 000000333000666020728 111111000444219 12 oEorr B 0030633 ica 1% F $M7e8a7n Group 12 735083 121917522 851 ia32l7 233548 2] 175 5 1 37 5108.83 1188.53.5 53.0 3150.137 a10l1s.5 B 0030609 11059 13 F Reference Range 4 158 2 a2 104 388 - 317s8- 8o- 06 - 230: Page 2 + 418-018:PAGE H-13 Client: gAracugsUsRegEIssNeECAaROCnRYc.PrOJRamAcTREATDORIES3,016H2e10.168BBa2t8sFaGSxel3R0l1s.Ee0t7Y7e-t0s43310/12/2000 (215) 443-8710 Ajescsice ession Spec. Gp SexAge GoHOeLEST gGLeUCsOSE LnDLs-DIR HGDL-uDIR TRIG Mean 73 136.4 2.8 33.2 133.8 418-018:PAGE H-14 ANIL FOTBa0usSI, BuAeIenNACsOdRaPfeOdRRTED | SonSS emmesoBateSh Rn 0USSoaEmer1/12/2000 CN. Tete Segeentn Sorters client vo. 1028 (215) 443-8710 stolys +10UIE DMS OS Spestent BAT fAocecseicon essionS3pec. | GpSexAge i CHOLEST GwLUCROSE IDaLp-eDIR HDeL-eDIR IoRnC, Fanny 1 f gi i802 PEER (i 2 #2 4 g = Group + Bay 1: gag 88 --_ rage 4+ 418-018:PAGEH-15 Chtents augus sIasNECNORCRHPOLRARACTRENTDORTES30,1-9I2W1-.0168DateFaxG:er30l1s-e97r7a-d0:433 2 -- AccessionSpec. El Gpsex`AgeCHOLEST GaLUlCOSE al a LDL-DIR HDL-DIR TRIG 418-018:PAGE H-16 ANI Clients 3B2A08RU55GI0ULB8SEu,NRRCREIEERSSNYEEACAAORRCCRHHpP1OLoRABuAOTREATDORIES30,1-9I2N1C-.0168bBpaaattteseFaRxRC:oeIlp30loS1er-ct9Et7e7edR-d:0:4330182//1272//22000010 Client Yo(.2151)028443-8710 Study: 416018 DANS 105 Species: RAT PEfacesil on o GOMES gues REI PIR a RG croup 8 gBoodoesses ddiioonr 22 B rogt J Reterence Range 2FA]I PBon a SP E ETea T1E y3o iin wo wo oose g- a: 418-018:PAGE H-17 INCORPORATED s0r6oones "Fan sorisrs0663 Client: JBBA8RSOoUGSE,sUESTRRARREEEESEE2ARSCHsEoLoAuBa-ORMTORIES, INC. BDaattee CSoLllMscSta"d: 12/12/2000 Date Reported: 04/17/2001 Client Yo(.2151)028443-8710 Study: 416016 DAMS 105 Species: RAT iRoosasaion3SBp'ee. 5pH pooompaeoE mtniaesy 10308 iieg pBoaoesEt ass seegan Group 1 GpPSeexx hAgaee ATRd,OSMPTAAL1R3E,ETI0L BFRhEESANomjw.n ERNEeyLbLEA 1 EF 11 ooff ThBpsEeee Togohesggoes 10 op.al1g bI1iRe3g5g.:a88 1ifof SF50aastsa S8%F ohee 0 6B TW5E7 ehaigele oTaso nLegess aonr BB5Booooaooomyvoceessssesteea iiH11eee0e1sse9 1BBB0 EEEEF ppBH ooooosenE ssesiri H nioieseass BI18 Ef BosdEss HS BF seeagn Group 10 BL0E.CeE8E8 imH79d.nB6e8 eHo0d.nd8OPpBIga 0.31 o8l gSgai3adf0s somfemererge gogeaEInl a ggnEEns d8oo3LlR2 Sig EE 88 ni 8 3T3h gTamms 3am Luay adnl Reference Range 33. 8so 8o- Ssh7- B13aPage 7+ 418-018:PAGEH-18 cident PET LA ETI wo. Tuts GOAT INCORPORATED 301-921-0168 Fax: 301-977-0433 1 sate Reported: 04/17/2001 (215) 443-8710 Ajescesaiconession Spec. GpsexAge TA 74, TOTAL RL EE T3,FTREO ETT 3 TASHL TFREEE T4 418-018:PAGE H-19 INCORPORATED 301.921.0168 Fax: 301-977-0433 Client:SA0R8GUSSuEReERSyEADRrCiHveLABORATORIES, INC. DDaattee RCeoclelievcetded:: 12/12/2000 HORSHAM, BA 19044- Date Reported: 04/17/2001 (215) 443-8710 Client No. 1028 Study: 418018 DAMS IDS Species: RAT Rho: ocSe pecs .|sGi pSo exAnge BBB 000000333000666273458 11111i0o0e85i75 111333 EEE BB 00003300663378 11i1c0ess 1133 FE M5ea87n Group 13 BD4E/MB0ET|AL1NG/3 DL. 10 FPRGE/TMELTA S 1L NgGH/ML__TNEGR/EOLTa 000:.:823033 TT7ea81l:.e9os88 0.76 o.ea 3.48 0.82 0.25 0.17 00::2866 8ee3l:s8 0 0:8 i1d% 9d1i%e 2d7e7 E Teaeay Eagsa F1.eRa 198 BBB 000000333000s85s33233 111000988253123 233 EF EF BB 00003300333358 1100063337 32 EF M5e7a87n Group 2 422.1328125 8s915.ld208s 800.7p853 32::338 a8f3:e261 Q8i 21..680883 e1a8.1a5e5s5 5132828 d01.3l0s4 1o1..l20f13 o2d%s 06lsd7 1Il.e9s62.334 BBB 0000003330005385357 111000998423398 333 E B 0030830 10841 3 MSeYan Group 3 00o.:00f80 838.865::783 o00.s86t%5 21g.i;1s2 809.g370 6:5 68:38 ols Gise 8131 C2 183" T2 es.753 L4ess 11 .228 2S7e Reference Range 73- 2s8o0- $0Page 9 + 7 57-129 - A18.018PAGE H-20 Client: ABBASROoUGRSnEUBRSSRtERRCIEEESNYEBCAaEORCRHS1PoOL0RAuBaA-OTREATDORIES30,1-9T2N1C-.0168DBDaaattteeeFaxRRC:oeRlp3S0lo1Eer-ctR97teY7ed-"d:0:4330142//1172//22000001 HORS; E8 0d Client vo. 1028 Study: 410018 DAMS TDS Species: RAT fReoesession S8pee. 5Bppooooowsmeevoireeisiigi1ies9sesa: BBooegesr diNoes steeagnn Grow an SexAggee TBAarTbOeTAL TR3e,jTo0LTA-L FBROEjEWLTNSej.un PNROEsEb.LFA fi1ffffFE 0ge8:a3a3z eSgar1nm8ae eS0Rm BogEmm 0oSsni ffff gean gahsd S8s maaR 88gB 2 Er rE eds es E Lass am 5BpBooooosssgoseseese0 r1ii0gd9st5ee7s 3332 }EOEF B Booo tseg r isse 22 ff seeagn Group 5 a L2n.a8g0asasge dense 1hn9.Et00 ns0n.5nin2 o0 00.8n97 E Boowin os 8 on o8la T5ow aWlagse e8sr sfie a3n% 5Boooj0sses 110098m 6 fFE Reference Range J 0a.080 250.371 s03k8 0u.m2 80.B21 3- se. 9. m- pa- 7 100 0 341 3s Page 10 + 418-018:PAGE H-21 INCORPORATED 301.021.0168 Fax:301-977-0433 (215) 443-8710 jccession spec. No. Ii op Sex Age Ta,TOTAL T3,IOTAL UG/DL NG/DL FREE T3 TSH PG/ML' NG/ML FREE T4 NG/DL --p 418-018:PAGE H-22 Client: JHagoT; nLsIeeEmnScy ODRasAoTraEtDoszesS, Rvc.SaBt3s8 nRSdelEeigaEteerr arnaseome Eg Re netetret esses Chtons wo. 2008 (215) 443-8710 gunigs vine THE We Spusievs AT rFAcoceassiorn Spaec. GpSexAge 1F4a TROTAL 1T3.hT0TAL FEREEE T T su E FREiErTA ppooemesmhsse 22 ooff ae 9 yr xeterence range egRoeBdE gm LL, 5BE HW OER CH WE 8 30 me 3c pn ae 418-018:PAGE H-23 INCORPORATED TE ae (215) 443-8710 301-921-0168 Fax: 301-977-0433 sete Reerted, oujser iB o1/16/200% ANocscession1SpBec. | Gp Sex Ag9e GSCeHIOoLLEST B 0031037 10906 1F 26 FE TTTTTTTTmTeees J 96 INCORPORATED 301-921-0168 Fax: 301-977-0433 AcNocwession SipBec.| GpPSex AAggee GHCHOOLBEE:ST TTT 2 418-018:PAGE H-25 ANI SH S300EsLuIEmETSoT RITEDE| RSoaSTBakLeSeSSL2R0saEevres12/20/2000 ETL poe pete Teporvet easterened cttens vor 1078 (215) 443-8710 guuty: some 108 NEE Spestes: maT T i R T7dTe ion CHRtE EFPIonuBmaaHiHiEEIaEEnEEi31(Ef:f 4:53{ ores 2 wweyan 3bo PciEomEme mmE {111% 3:; w2ean +b5e I HS p P 0 E I in E n 21(11 4#,2: geameing ff i orowp 5 fs i Reterente Range soe --. S18-0I8:PAGE H-26 Client: AJR3GCUUSS, RRIEESNREECAAORSCRRHPYOELARBAOTRAETDORIES30,1-9I2N1C-.0168BDiatteeFaxSC:oLl3U0l1Ee-cE9t7Y7e-"d0:43312/24/2000 BBO18R)EAaS4aBTy8nad1o0ss - Daattee RePpeorteodt:: 01//21e6l/2001 Crient No. 1028 Study: 418018 DS MILK Species: RAT hAecscessions8pec. GpSexAge cSHEOTLHEEST 5BBoopaoio esne ei 10n3718ffkfEF 32$ segagn riel Group BBooostss 1i0e99s5 77%EF iaa seapnr B5a Group 7 5BBBoooonnosniiittoeeaar iH1h0ns9ga9ses f8 o:oEokk 32i4 BSBBoooeoooasssseee mnniisesgeese:l 88 koEkF pense ns 8 ok i32S 5 Reference Range ss - 9 96 Page 4 + TTT 418-018:PAGE H-27 Client: ABmScusRRIEESNECAORBCRRHPIEOLARBAOTRAETDORIES3,015I2N1C0.166DBaatEeSFaCRxEoZl30Al1Ts.Sc5E7t7Ee.Td0:48812/24/2000 BTORREEATSiBnaad1004s - Datee ReppeorRtepeordt:ed: 01/s1168//220001 Client No. 1028 Study: 416018 LDS MILK species: RAT hAececessions8pec. GpsexAge gSrEo7r5eEst B 0031070 1K1oe0ga08n 8 F i8:s127 Group TTT ppB8 oeoeo3ninmom7y 1nn n1e0d0e0 33 OfifF 1330i3 BBBBoOoonRneIEesS nntisde 53 kffOFFF 13i33 seeagn 5 3 Group Reference Range 12 - ApPTpOroved ------=H-= SSl/oe Dattee -m-mmmnthii!di/ 418-018:PAGE H-28 ELA TID Bsns den eves BH fy JAosecsiccn ession gSpec. | Gp SexAge EH GHOLEST Gn GLUCOSE IDwLa-eDIR HmDoL-eDIR TmRIEG 418-018:PAGE H-29 INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 Ajcecescaionession spec. Gp Sex Age TCHOELEST GGLUnCOSE WLDLa-DIR EHDnL-eDIR mTERIG ae 96 319 0 125 418-018:PAGE H-30 (07)0(O4S Client: A2R3SGUS8 RRIEESRNEECAORDRCRHIPVOELRABAOTREATDORIES30,1-9I2N1C-.0168BDaatteeFaxRC:o2l3L0lE1e-Vc9E7t7Ee-Td04:3312/08/2000 H(O21R8E)RAN4S43B-a87201004s - Date RepPoorrtteedd:: 01/16/2001 Client No. 1028 Study: 410016 PUPS GD21 Species: RAT hfececession s9pec. 5BBB o888a83333330000383833333388 1110000538533353028 BBBB 8888308333330068855833388898 Tf1o0oe5o35ki7 165i soenan Group 3 Gp Sex Age CSHAOTBLESETST GLSUECSOESE LRDULA-RTDNIR HMDEL-RDEIR TRWITSG: 3333 75H8l1 758538 i38 i 1ii7e 18 F3H31 38 333 3 18888 8 ii5% % 3i&%f 8 #i#8 iB #338 38 a #1;Tene a8a 13 61 3i18 5BB 000000333000233433133 11To00s98i88?85 B 8030533 Toads 4 BBBb 888830033333000330313s258 111000033333380 10538 44 4 seoarn Group 4 a%&s 5 a5s8i 58 iEEsod 3 13i3e i 3i31 3 7#i% 8 337889 3% 83i% iiiet 58 i 3#3 i $#13 TBids ies 1055 285.1 Reference Range 388 -7[.ON Page 3+ 0-0 8 -3125 AISOIEPAGE HA clients ARERVGSUSESRIESNSECVORTCREPEOIRABAOTREAIDORIES30,1-9T2H1e-.01683a3t5e2FaxSS: e3El01i-So97r7E-a0d433Laon 2000 (215) 443-8710 Ajecacnession spec. Gp sex`Age CHEOLoEST GR GLUCOSE Ee LDL-DIR HDL-DIR TBRIeG 418-018:PAGE H-32 CLYTICS civ sro sn 0 sans wo or INCORPORATED 301.9210168 Fax. 301-977-0433 Client: A9R0G5USSHEREEHSYEARDCRHIVELABORATORIES, INC. DDaattee RCoelcleeicvteedd:: 12/08/2000 BH(OU2RI1S5L)HDAIMN4,G43A-P8A71019044- Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT AHescceSs5pesc.i|oGnp Sex Ade BBB 000000333000332687e5 111000399888785 777 BBB 00000033300033376s09 111000999895910 777 BB 00003300337721 1100889923 77 M5e5a7n Group 7 HCGHJOOLLES|T GMLGU/CDOLSE IDMGL/-DDLIR HDMGL/-DDLI=R TMmGi/gDL 815 757407 i25 11i87 233827 2385 Ee8ls8 2322 ii3ss 222530 558 588 33 ii 3336 s$a0.5 710033 T8als 115a 350 Reference Range 3488 - 317s8- 80- 80- 3125 Page 5 + AISOISPAGE H.33 CH J a EAPETTBaamaHLteRegniusm,e (215) 443-8710 jAegecseican ession 3Spec. Gp Sex Age Re a T4,TOTALT3,TOTALFREE T3 TSH FREE T4 Mean 0 Taz Tos 418-018:PAGE H-34 GLYTICS sociussion se 0 anos vo zor Client: ARGUS RIESNECAORCRHPOLRABAOTREATDORIES3,01.I9N2C1.0188DateFaxC.ol30l1e-c97t7e-d0:433 505 SuEERY DRIVE Date Received: 12/08/2000 ORERAN; Pa 19044- Date Reported: 01/16/2001 (215) 443-8710 ip : Client No. 1028 Study: 418018 PUPS GD21 Species: RAT NAeccession Spec. BBB 00000033300032388831 11111100085%43 BBB 00000033300022288845 111111000686280 BB 00003300228878 1i1c0e6s3 Msea7n Group 13 Gp SexAge 111333 111333 1133 1U4G/TDOLTAL 73NG./TD0LTAL FPRGE/EML13 08g::90o90 alie o08l:0g90a8 29illss8e00 g0g0a 90lloo00 80 i52a8 JNSGH/ML 1.16 T5 E FNRGE/DELTA 897:3080 008.:1000000 T0o BBB 0000003330002222275 111000909111756 222 9gg.go0aa0 800.ll4oo3n0 0.07 BBB 000000333000333233890 111000888113ss0 222 08i:0308 BB 00003300323312 1100982388 22 00.:0800 M5e8a7n os 2i.s8e1 K8 I Group 2 Reference Range 73- s2000- s0Page 7 + 25a7-1a98 - 418-018:PAGE H-35 SRL ALLSLL aC Aho INCORPORATED 301-921-0168 Fax: 301-977-0433 H(O2R1S5H)AM4,43P-A8_71019044- Date Reported: 01/16/2001 jJocsiacnessionspec. op Sex Age EEE T4,TOTALT3,TOTALFREET TSH FREE Td 418-013:PAGE H-36 CAYTICS sve st 0, sso vo sr INCORPORATED 3019210168 Fax 301.977.0433 Client: A5R0G5USSHEREEHSYEARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 12/08/2000 BHOURTSLHOAIMN,G APA 19044- Date Reported: 01/16/2001 (215) 443-8710 Client No. 1028 Study: 418018 PUPS GD21 Species: RAT + Aocession Spe|cG.p Sex Age No. I 74 10TAL UG/DL 73 TOTALFREE 75 NG/DL PG/ML TSH NG/ML FREE TA __ NG/DL BBB 000000333000222543051 111000999565908 535 000.1:000000" 6.00 BB 00003300223233 1100998631 33 B 0030334 10963 3 888:::880000 823.:l38g698 0.00 0.05 0.04 BB 0000330023835 1100596653 55 06:0000 20:0001 0.00 6:02 eS7a0n 0 T3i6a 98 "R0i3t7) Group 5 BBB 000000333000232588789 111000998777425 66 BBB 0000003330003326130 111000998787806 666 BB 00003300336633 1100598821 6 M5e8an Group 6 00:.0060 03.:0206 0.00 0.00 0891i.809000 0.00 0.04 9.00 00::0388 09.:680 _ 0.00 00::0000 8o iTe3989 8"04 8o Reference Range 73- 2s000- 60Page 9 + 25a7-1a9d = IBOISPAGE HAT AYTICS 200 Girars Sree, Sute 200. Gathers, MO 20977 Clients ZBy3o6EEu8sURfSRNeEeOIseNeCEaOnRcPnOP RATEDT301-9o21e-0168BRsaa2tt2seFaxSSR:eEa3p0Lto1-Mro9t7Ere7R-d0i4331001//0106//22000001 cient vo(.2151)026443-8710 svar: eteots PUPS anor species: maT RemasterSakpee Ee R ETLR RRL B IERaERenA EjS ohopnasneaeotlanay] s] S pMrB eeow ow oon 0.00 |F PilnE easmE faeit 7 $0.800 one 0.00 croup 7 porrr 5 OE 3 5 seterence Range s3 e sBeee 8o e SsWe BaEs- coved <cneran ile eemene pate --eeildidis pp Page 10 of 10 418-018:PAGE H-38 Chient: pynseugsusRzxIeceNeECaaOsscRcyPnOTRARAoTREATDORI3E0S1,-92I1n-e0.168BgaagEeFaxES:oRt30iA1t-e9S7c7eE-d0s43310/12/2000 EERE a sate Reported: 01/16/2001 (215) 443-8710 ANocscessioniSpBec. | GpPSSeexxAAggee CCHHOOLLESETST GLGLU7ECROSESE LDHRLoR-TMDTMIR HEDRLLo-DTMDRIR TRERIIGS. srs 32 IB0ISPAGE 130 ANI Client: BA3ShR0ecsItSsREoSeIEESNyESCEAORCRoLHPOLoRABAOTREATDORIES30,1-9T2N1C-.0168DBDaaattteesFaxRGS:eopl30oSi1rs-to97eEt7da-:d0s4330110//1162//22000010 cltent vo(.2151)028443-8710 study: 418018 UPS D5 Species: RAT Sfeloen ton EeGe l eela l ma Tam mH TE wm BBohsbetuHso nhtf Benes ue i BBoomB BoEa x 08B Bo B aroup 31 orou 12 croup 13 xo OFF OF sage 2+ 418-018:PAGE H-40 CAYTICS INCORPORATED coms su ss 20 03m v0 2 3019210168 Fax: 301-977-0433 Client: AB$R0U5GIULSDSIHRNEGEESHEYAARDCRHIVELABORATORIES, INC. ORSHAN, PA 19044- DDaattee RCeoclelievcetde:d: 12/12/2000 Date Reported: 01/16/2001 (215) 443-8710 Client No. 1028 Study: 418018 PUPS IDS Species: RAT fiReceS3psec.s|iGopSnex Age GSHEOJBLESETST 5BB 000000333000668333738 111000939333331 222 B 0030638 10938 2 i31i28s8 iis B 0030630 1937 2 17 e7an 152.07 Group 2 BGELJUOLCoO-SE IMDeL/-oDnIR 12783837 32a12 32332 328 819%5".2 384.2 EMDoLo-nD.IR TRMGI/GBL 33383 223011s83 3232 18731 313.88 81885%.4 BBB 000000333000684d41z3 111000959243328 333 e58a0n Group 3 12i30s82i 232806437 k8i4]7 4332 233337321 8 167) 2B88h) 5B2e d31h 3l12e BBB 00000033300068444645 11100099956677s 533 BBB 0000003330008864i47s8 11100093368%88 355 M5e7a8n Group 5 13228 i23s7872 7 283 22232 28 22280 3 227181843539 2184d 3B6i3 201 3128 18313 E18y.7 z8e580.2 519% 280%.7 T23874883 Reference Range 6so - 317s8- 0 - 80- 3125Page 3 + S15.018:PAGE H-41 ANI AINYCOTRPOIRACTESD 230010-9G2a1r-d01S68r. SFuaixt:e 230001,.6G7a7i-th0e4r3s3burg, MO 20877 Client: BABRoORGUESSa ERREESEByARCHss0LaAsB-ORATORIES, INC. BDiattee CSoRlLleVcEteRd: 12/12/2000 Date Reported: 01/16/2001 Client wo(.2151)028443-8710 Study: 418018 PUPS 105 Species: RAT RheeeeseionS 8 eSel xhae a GSEiBoELSTaet GSEiUSgS"er B35 oo0o0%33000e8a5e00 111008381707 8 n FCT ow3% seseagn mBoYE iso Group B5 ooossoeesy 110890%5 77 seeagn Group 7 0Bm20 BELo. #Ho0o 385Bg0ooaooi03xg08eeesssssss 111109110898%80 8 822Bog00o3ii300ee0c2ess8p iHHiiigtsgese: 88 5083883 11888 8 Reference Range un n e og ow aBn # 3 moo 1 a8 i ise. Hme- rage 4 + IMDRiFobn mMRoRn GEeES,. E=H8 4 4FAB TT5 THhs. 35 EBea 23 8a 532 T8Es. W 8E0E% FF87HH] F8 3FsE I 3] 8ze 3333 a n F# H g s 8 i nH o8 - 08 -3:13 418-018:PAGE H-42 INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 Ajescesaiconessionspec Gp SexAge CHOLEST GaLUClOSElLDlL-DIR HDL-DIR TRIG 418-018:PAGE H-43 ctdente JPagIsLEE IELAPAA CAONIESD, Toe. BJe5r8ewBSEHInApTELTaE sarszsamee EEE on sate Reported 01/26/2008 (215) 443-8710 AJcciesasinongspec. Gp Sex Age a T4,TOTAL a T3,TOTALFREE T3 TSH FREE T4 418-018:PAGE H-44. ANI Client: ARGUS RIESNECAORCRHPOLRABAOTREATDORIES50,1.9I2N1C..0188DateFaxC:ol30l1e-c8t77e-d0:433 H9BO0UR5ISLHSDAHIMEN,EGHYABADRI1V9E044- Date Received: 12/12/2000 Date Reported: 01/16/2001 (215) 443-8720 Client No. 1028 Study: 418018 PUPS LDS Species: RAT nAeccession Spec. BBB 0000003330006se8d32 111111000332315 BBB 00000033300066688s75e 111111000333453 BBB 00000033300066898s08 11111100033378 M5e8an Group 11 Gp SexAge 1G4/TDOLTAL 7N3G/DTL0.TAL PFRGE/EML1-3_ NTsGu/ML ii1II1 8g0:i73g000s 1a30r31i.:ea58s 3g9l.aG0d0 1ii1II 9g.i9o00 i1iII1 888:::000000 333303:.1465298 I0.G0S0 80 2527..536888 0030%6 NFGR/EEDLTA g80l:.o00101 000::.000300 o"gf0?8 B 0030691 11049 12 0.00 42.35 0.02 e5a8n 80 o12.35 8"oz Group 12 B 0030692 11059 13 0.00 40.17 0.15 0.01 e$7a5n7 06 1o0.17 635) 0"ot Group 13 Reference Range 73- s2600]- 60Page 7 + E5XT7ON. 1.9 - 418-018:PAGE H-45 ANI INCORPORATED 301-921-0168 Fax: 301.977.0433 Client: A28R8GUSSarREmSaEARDCiHveLABORATORIES, INC. DDaattee RCoelcleeicvteedd:: 12/12/2000 HORSHAM, PA 19044- Date Reported: 01/16/2001 (215) 443-8710 Client No. 1028 Study: 418018 PUPS LDS Species: RAT Aocscession Sip)ec. | BBB 000000333000888333738 111000889222331 BB 00003300663490 1100882378 M5e8an Group 2 GpPpSSeexxAAgdee 222 22 TT4ERTTOTAL TR3,TbOTAL 00o:8l21s3 193274..137030 00:a57 8'38i1s%d T12i2s7s TSa3i.ahz FBRoEEj103 o0o1l.8ss81 l.s437 TNSeHjML oss 0.28 3) FRNEe/EbLTA Fl0:Eo1Ts8 38:10180 "1o0s36 BBB 000000333000668444132 111000009432289 333 MSe5a7n Group 3 000..:000000 352583:.:168792 9 7208.863333 0.68 00..0010 8"es 1T0000375 BB B 00003300664445 003084e 110099567 10967 55 2 BBB 000000333000886444785 111000935667890 332 e7a8n Group 5 000.4:2080 1980072.335118 08.1d0 1a 000:.300186 000:11230210 8e6324l11d2%s8 00::0083 T2392337 T177e..1a7g8s 2374 51738 1T0o2s7i Reference Range 73- 3s000- sgPage 8 + 257h. 21.39 - 418-018:PAGE H-46 GLYTICS ovis se 20 specs vo or INCORPORATED 3015210188 Fax 3019770633 Client: BA8RU0G5RULSASIHNREGEESRAEARDCRHIVELABORATORIES, INC. DDaattee CReoclelievcetde:d: 12/12/2000 HHOoRrSeH)AMt,ha B8A71d19044- Date Reported: 01/16/2001 Client No. 1028 Study: 418018 PUPS LDS Species: RAT JAi ccession S3pec. | 5BB 000000333000668333270 11100083877378 M5ea0n Group 6 GpSexAge 1T4E//TB0ETA-L M73G/TDOLTAL FPROEEM13 TNSoHjML__|FNRGE/EBL14 66 008.::000000" i33408:l:8s38s0 0.00 5.00 05.:0003 60 D0o.7825 8 s0 06 BB 00003300663533 1100395975 77 M5%ean Group 7 0.00 25.50 oS 2s5.5 BBB 00000033300066633575 111110009009079 888 BBB 000000333000666338930 111111000088432 88& BBB 0000003330006666e;31 111111000000556 & Reference Range 808.::000000 880:::000900 008:::000008 33- 843938.::635783 7ai3ds 34ds7a.:93d88 1s8o0- 0I.G0E0 93.:0808 S0- 0.01 088.::088311 800.::08822 i5a7-1298 - Page 9 + 418-018:PAGE H-47 ANI Client: pSa0ugu5EsUSSEuRpIEhSaNECAORER.POLRADACTREATDCRIES30,1-9T2H1C-.0168BBaatteeFaxCS:o3Rl01l-Le97tS7-t0:4331/12/2000 HORSHAM, PA_ 19044- Date Reported: 01/16/2001 Caton vo(n2152)020443-8710 Study: 416010 PUPS 05 Spectes: maT RjoceesisoneRaion pee en Ae RBRGTR JRRae EFRELT TMgeEBoR h Fosase 1w1e0p5r 33yy wHihn y To E2S:2o]s aroup 8 BT oaobbomdap neEessEs hioonnnle 2233 gE g0g0e E Egmaiys ooam 380.]%05 Boooobhhoiaioggseeeeidsness hiiaoooysn 2223 gg[gee1e ai LEaEEe, i02.g080 oywr 3i rwior g iBn croup 9 Reference Range 33 - s2e g$ e sro 113- ApproveBosme o THmeTmeememnane Date --eidlinlil ANTE 418-018:PAGE H-48. INCORPORATED 301-921-0168 800-237-2815 OA AUDIT REPORT TGhoeod rLeapboorrtatloirsytePdrabcetliocweswaasndrewviitehweEdPAfoGroocdomLpalbioarnacteorwyitPhracttheiceFsD.A Tahcecurfaicnyal arnedporctonsainsdteanlcly asasnodciattheed frianwdidnagtsa wweerree rreevpioerwteedd fotro management. Tahcecurmaettehloydsrefulseecdts wtehree datthae. meTthheordesforde,esctrhiebseed staunddiesthweererepdoornte in compliance with the FDA and EPA Good Laboratory Practices. Gtdae Franklin B. Newnan QA Auditor sponsor: ARES RESEARCH LABS stopy: 4/60F REPORT TYPE: HEH, 73T,H, TSH FT3, FTH, ctor AUDIT DATE: 1-3-0! REPORT TO MANAGEMENT: 1222-0) AUDIT #: 0- 0L3 3h He DATE SUBMITTED TO CLIENT Te 0d Ragwind Veer fed MANAGEMENT 1/%2// HISOISPAGE HAY ANI Client: A3R8GSUSSHERRIERSNYECAORDCRRHIPVOELRABAOTREIDORIE3S0,1-9I2N1C-0.168BDaatteeFaxRC:eocl3e0lf1eV-c9et7d7et-d'0:43302/01/2001 Client voHB .OR1A0a 2N8: Pn a 1004S4tudy: Date 416018 105 WILK Reppoorrtteedd:: Species: 04/17//2001 TAT FREo 1B PSex Age CHOKE: BpgBB eo0oep0g3cnn5mh52iiso9 innn11n5ige0a1 11111 koooFfff 13ii6825 TTT BBBBooooanoaniasoimnnise:eel 1111 ooootfff i2i2 ppBpooooonmmsosesniniiiinne 1111 :ooofff i582 pemsiinn 1} i3 sseeagn 152% Group 1 BHe5 mooEoeiasiEesoos 11620 teens 10 E 11gg ft iF20d ppoooedssdselies Heena 111gg :ff 22a2 Reference Range 4 he 96 Page 1+ 413-013PAGE H-50 INCORPORATED 30162-0168 Fax 2019770630 Client: BAH 3N5OG0RUESi SASEREESEBAaRBCRH1E0L0A4B4ORATORIES, INC. BDiattee SCRoElElSeEcEtYe"d: 02/01/2001 Date Reported: 04/17/2001 (215) 443-8710 Client No. 1026 Study: 418018 LDS MILK Species: RAT AHoc:cession SiBpec. | Gp Sex Ag9e GHHIOLDEEST 5ER o03sels 11638I 10 2581 Heogan 31854 Group 10 pD5 ooooe3sstee1as 1mn1i8ee3s0 Bose ne ii11iE:EF ik a%8i ppBpooooasodeeseesel ennEeseed iiiikk:k 182533 Bosses Hee if i sooepan 5Fe0] Group 11 5pB0oe3sdaeaes7 1ineee4i0 13 EF BF 288 Reference Range g8 - Page 2 + 418-018:PAGEH-51 Clients APERELGUrSSSIEESNTCAEORGRHRPOLRABAOTREATDORIES30,1-9T2G1-.01683oB2ea5st8seFaxSSS:aeSl3rL0im1Igr-eS9et7Ha7s-Ttd0:4330mao/poin/e2e0n0 (215) 443-8710 Acoessionspec. GpSex Age TemoresT TTT J 96 418-018:PAGE H-52 INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 FNooscession S7mBee. GppSSeexx AAggee gCHOOLLEE:ST TTT wean i #2 FEAT RS ye sana (218) 443-8710 NRoowcessionS2pB'ec. | OpPSex Ag9e CHHEOLLEST TTT oe 26 418-018:PAGE H-54 GLYTICS coco sn 0 canna wo sor INCORPORATED 301-621-0168 Fax: 301.977.0433 Client: BA5UR0T5GLUDSSIHNERGEEHSAYEARDCRHIVELABORATORIES, INC. DDaattee RCoelcleeicvteedd:: 02/01/2001 HORSHAM, PA 19044- Date Reported: 04/17/2001 (215) 443-8710 Client No. 1028 Study: 418018 LDS MILK Species: RAT RAec:cessionS8pec. | GpPSSeexxAAggee CGHHO7LBEEST TT BBB 000000333333532788901 11111155377785 FEFEEF 112E37d BBB 000000333833333888334 111111533788920 & FFFEEE i1180e38 BB 00003333338856 i11d5e8s4 EFF 10523 es7aBn 2170.78.8 Group 6 BBB 000000333353353888878 111111333986237 777 FEFEF a776 MSe5a7n 62100153 Group 7 BBB 000000333353353999201 111111666000220 888 FFE Reference Range 13745 74 4586 - Page 6 + 418-018:PAGE H-55 Client: ABABReoUGSBUSiRREREIESSNEECAAORRCRHCPHOLRABAOTREATDORIES30,1-9T2N1C-.0168DBaatteeFaxSC:oLl30lM1e-c9St77e"-d0:43302/01/2001 BSRERRES Ba 1o0us- Date Reported: 04/17/2001 Crient No(.2151)028443-8710 Study: 418018 105 MILK Species: RAT RheeseesslonSpe. GpPSeexx haAgee gChHoORREEerTTT P BBBoooomoey asd nHHeer et: o 32fofff 1i1a28s82 5pBoogooEiEedssse HHHeses f8fffof iiie yeeagn firen | Group 8 BBBbooooodosddseeedddo eef11e6nn1r1 3g33 fEff 1i933 pg55oooogdotidieeeedsdniee HHHHeeeettll: 3f53ofkf 143538 Bode He 3 of 1% seegan T18o%s Grow 3 Reference Range o 4- 96 ApprovedSage O Tor S 7 pate 2c 418-018:PAGE H-56 INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 jAoecmaiconession spec. Gp Sex Age CGHOrLeEST GLgUiCeOsSE LDpLo-oDIR HgDoLo-DnIR TRgIoGp 418-018:PAGE H-57 ANI Client: BHANRESG,UESa,TEEyIESNLEACEROCRHRoPOLoARRnAGTRAETDORIE3S01,-92T1N-C0.168DBpaaettteeeFaxGsS:oEal30Rlp1sS-ec9Etr77eYe-d0:433eojyosrm/y2z0emnn ctens wo 2028 (215) 443-8710 study: 410010 oaws 105 Species: maT --Racasaiongee. TG Sex hae GGloLLEger guGeIEoSer guoarle pgp rtp gpusi, arowp 12 pommme nse 2 f BBooontdees sshle 11 ff zg = 8$8 == 1 209 :PoE8 8Zz oroup 12 9 96 319 80 104 418-018:PAGE H-58 CLYTICS oars sie. ss 0 asso wo or INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 fiAccession3SBp'ec. | Gp?SexAggee CCHROOBLEESTST GLSEU7BCLSOTSE LMDOLJ-oDbI-R HDMeLy-oDbIR TRmeIjGbL 418-018:PAGE H-59 GLYTICS INCORPORATED o301c521c:01s68 e sFaex. 02019o7-n060e3 s vo mm Client: A20R8GUSSHERSERSYEARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/02/2001 EORSHAN, BA 19044- Date Reported: 02/13/2001 (215) 443-8710 Client No. 1028 Study: 418018 DAMS LDS Species: RAT A NoscceSp9 esc.siGop Snex Age BBB 000000333235777888987 111111335000321 I11 EEFFE BBB 000000333528777898301 11111125500086s I11 EF EF BBB 000000333335777888533 111111335000887 111 FE F BBB 00000033322377798387 111111388113210 111 EF BB 00003382789090 1111281133 I1 EFF 5e8an Group 1 1NG3/,DTLOTAL w357a.067d47 2ss3e0sl0 17s783l8s13l S7Soi8ll4ss3ie 8553058i e1753861s 1U3G,/TD0LTAL 2G1l.7a3s0 11 21133%88 ooilldeesss 01g11l93s35s Ttei2asn 1N8O/3ML___ m91:.s86s18 2IL:f3Ee5 2L338g3 2101..047337 aoiiee Tlaites"s FPRGE/EML1&3 FNRGE/EDLTH il0.o4da3 100..5A682 0008%%7 0807.907 00ois8s o1Gli6ad3d 00013838% 00oill8%s38 o01l8s% 00.i7@7d s13a8s 7geizt BBB 00o00o33323s33s00o38l 111111666333830 ii1II1 EF BBBo00o00333s33s83000d88 1iiileee3s:e IiiIil F B 0033807 11637 II F Reference Range 3i2767:.758582 iias6slloa7sla 1 2s000- oo0l.le3es7 ggollooestss or 7 3- i28.3e5 11o:3s38s3 38 s57a 0o0:13i88 0 o0i1l5s 858 30- o00i.f32p08 000il333888 oi 21:.39 - Page 4 + 418-018:PAGE H-60 ANI Cltont: g32 3B 50u508gsteyReeSIt EsNENeCEOnER--RcPnOp-RASACTAETDORIES3,01-9T2H1e-.0168gBguaatgteeeFaxSoR:eSlp3Hi0oI1rv-St9Eie7E7da-Y:0:4330o2z/y0i2z/y2z0e0nm Cltent Ho. 1028 FO FFe-- ein gee ekille AakTaR RRS TREE SFREEEE IE H uf EI ma Tsen eoeem e sw non yEoSun SgaEn TTHwE OLLEE E 3 OECh [-- BEEpom RNsolime EBmloUUnn UUusEEs HHBBBGBRIEP:: 00 BHmE3EBEa ror$OrL@m8eE Ip$I1ynRBm ppBO1He8RE oBii9dRd1B IBPFFiRooaeEmmEmnemneNedaGleE H2n#Emr:TP: gf HaESR fo28pef InirnfiRe EsIipEgEg iObpohHhdp FEFeEemEmaImNngeEe HPoozr @AmFmu PooEoh OanEyEE IggRny i4oBf Group 12 y5on E WaEETH aOWeE WTmm HTEa Reterence Range wBoe?3- PgA1- 8- 1n21wage 5+ AISOISPARG6E1 Clients L agus I EIaSNECAE ORCREPOTARBAOTRNETDORTES3,01-9T2C1-.0168D13a8t8eFaxSS:eEl3R0l1Is-cT97tE7a-Yd0:433cajonseonn DEtEsAoe sve Sapam SET cttenn vo. 1028 ovatys o16018 TS 105 species: TAT a Jcoessionsec. op TA Sex AgeT3,TOTAL pT4,tTOTALTE SH fF eR T3 FRfE EaEnTE 4 PFP geeesdmmde eilnieaasmnhhbEob r a daEEEgoBEorHRn InpBm4E ggggdann oeoenfnl ee croup 13 Th HE Le IE Ih fT7. 2/3000 418-018:PAGE H-62 Clients BBUSsEaEISNECAORGRHPOTARRAOTRAETDORIE3S01,-92n1e-.0168BBa5t8eFaESx:el3L0l1g-Ie9t7T7a-d0:433cajoarmom i vate Reported 02/13/7001 (215) 443-8710 AJoecmacn essionSpec. | GpSex Age CHEOLlEST GLaUClOSE al ak LDL-DIR HDL-DIR TRIG 418-018:PAGE H-63 CLYTICS oo ccvesion som 0 casas vo wor INCORPORATED 301.9210168 Fax: 301-977-0433 Client: ASR0G8USSuEReEnSyEARpCrHovLeABORATORIES, INC. DDaattee RCoelcleeicvteedd:: 02/02/2001 HORSHAM, PA 19044- Date Reported: 02/13/2001 (215) 443-8710 Client No. 1028 Study: 418018 DAMS IDS Species: RAT rHooscession Sbpi)ec. BB 0000333884478 111852%7 M5e8an Group 2 GpSexAg9e 22 E CGHCOJLDELS=T GLMUG/CDOLS=E LMDGL/-DDLI=R HMDGL/-DDLIR TMRGI/GDL 8ss 1387 31 750 25732 8B3 e 1W5H1 42a 721:78s s2l1 BBB 000000333533888453001 111111583233020 333 EEEF BBB 0000003332336683333;4 111111888333543 333 EEEFF BBB 000000333233888333756 111111233333678 333 EEEF BB B 0000333s883308 0033880 1111323480 11241 33 3 EE E BB 00003333886612 11113844:3 33 e58a0n Group 3 777% 132i82s45e 522 6&83 13T0a3s 7855 iiises iii 8E&2A 2018s81 8182 114858s8 15z E22H8 1688709 128758 311370s2 85 7p04s1 1881s0 E7S 118835 33 382 13E0] f7i.4 a182l.5 32.9 35%5!.6 1d0e1i8 Reference Range a568- 3197)a- 0Page 2 + 280- 130-2 AIBOISPAGE 164 Client: JugUS RIESEAERCH SLABOGRATORIESR, SHe.EDateieGol3lssceaade: 905 SHEEHY DRIVE Date Received: 02/02/2001 HORSHAM, PA_ 19044- (215) 443-8710 Date Reported: 02/13/2001 -- jccesaionSpec. | Gp SexAge GHGOOLREEST GLgUeCsOSE LiDhL-nDIR HgDiLb-iDIR TpRIeG 418-018:PAGE H-65 L 388 BL eteINCORPORATED HORSHAM, PA_ 19044- (215) 443-8710 EL 301-921-0168 Fax: 301-977-0433 Date Reported: 02/13/2001 ANcoscession S1pBec. | GpPSSeexxAAggee CCHHOOLLPSETST GSLEyUsCeoOnSE LMDOL/-oDLIR HDHeLr-pD.IR TWRMIoIGSL 418-018:PAGE H-66 ANI& Client: BayaEscubsmesLeIegfsNtENCaagORcRtyPOpaRsAoTRAETDORIES30,1-6T2N1C-.0168sBBeaettteeeFaxRSc:eoRpl3A0ai1Mrg-tcS97etE7da-Esd0:4330o2a//1a3a//2a000m1n ciient vo. 2028 (215) 443-8710 seueys +ieous ome 105 Species: maT SESE foeee aemmeaumnE la 1 RSE ogg oga wEE#T Baaa47 BeaeT nBw =e #8 jpi Fpgaaaaesgdmh iaiiinmnaeng1BgogEo of:ooff gggg$Ba oo0@B8# 0# 8 4 1111 oi8#H 8F8 B48i= pg ppaoEaana madhinia nnnilEigt gg1ggoo of}d g#OgN 0# # #8 1 1iER 44 oiH Z# z 4 croup 7 yoerar pallOiRaRl Bral Ed ppfPpaomaadmnisiiiunnlnmem pala 111f1 }fEf fo w2g o00mW@m #0 r1 14fo% gxioe 8oa=Fm 18 pRetearencue Ralnge it 2 # nBneo# s80 o+ mBeT a 4 rage 5 + 18.018:PAGE H-67 Client: ASRGEUSSREIESNECAROBCRHEPOLARBAOTREMODRIES3,01-9I2N1C-.0168BpaateeFaxSC:oSl3E0lL1g-Ic9St7E7a-Ed0:43302/02/2000 BOHRA Bh ssou- Date Reported: 02/13/2001 Client vo(.2151)028443-8710 Study: 418018 DAMS IDS Species: RAT feRoeasaionS%pee. BBooogompEiysnsieiaiz1l1euslessr wteagn Grow GpPSSeexxAgAdee GCoOLLEESTSTGLGLUUCcOSeEsE LLpDILGRpIRin EIRDVLSRDRIA IIMEIGG. 2B Eof FL= LE n. 33: 257 55i sGes mBoWs 1t7i e#e8 58ai pBp5 ooooaoosmsssgmteizzsis nidneetilnees 3333 oF pHppEooooosmssesmsiss ielitl nell 3ff 2 of gem HE ff wBeeagn Grow 3 [8 8 sIo 1a 31m ii$. a$3s0s au3I8s g 883 isut 4iii a33gig18g2 i 1 3 ji] HH 23 TWoea 31a smere ia Reterence Range iwo 7 M-89-9Fi.] 31-1 noprovegSuzm e Too r e pate HALL 418-018:PAGE H-68 Chtents $5GUS, EEIRNEACROCRH,POARRGARTAETDORIES3,01-9W2e1-.0168DateFaxS:el30l1s-c97t7a-d0:433 (215) 443-8710 Fg jccessionspec. Gp SexAge GCoHLOELTEST gGiLcUoCsOSE LjDLi-bDIR HgDLu-iDIR TmRIgG 418-018:PAGE H-69 INCORPORATED 30121-0168 Fax: 301-977-0433 Client: ASR05GUS"SHREEESHEYARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/02/2001 HBOURISLRDAIMN,G APA 19044 Date Reported: 02/13/2001 (215) 443-8710 Client No. 1028 Study: 418018 PUPS LDS Species: RAT ANeccession Spec. | GpSexAge BBB000000333533777444233 11111165660/34>38"/7664521 111223 M5e8an Group 12 CMHGO/LLEST GMLGU/CLO.SE 11315535 222522378 1w3a0t.4 21870.84 LMOGL/-DDLIR 23530 303.2 [MDGL/DDLI|R ZMRG/DL i33s8 2115882s3 73058.2 25375..58 BB 00003353774456 1111665546/655 1133 M5e8an Group 13 110732 222127 831 3323 223585 8137s 52133.5 8551.5 335%.5 i245h Reference Range 856 -7431.s 30Page 2 + 820-3 tas 418-018:PAGE H-70 33008, SETH. INCORPORATED EERE BvIe SmaAoNrEi cToopyees 301-921-0168 Fax: 301-977-0433 ade ver ooze (215) 443-8710 suudy: 413018 SPS 105 Spectes: BAT AJescsice ession sSpec. | GpSexAge Ea 13,TOTAL al T4,TOTALTSH FREE 73 FREE T4 18.018 PAGE HT1 CAYTICS socio ste 1 cso wo zm Client: AB22RS0ES,GUESSSuRmREEISNAEBCAnORTCRHjPSoOoLsARaBA-OTRAETDORIES3,01-9I2N1C-.0168BDDaaattteeeFaxRRC:eoMpl30olS1re-tc9Ee7t7de"-:d0:4330022//0123//22000011 HORSE Client No. 1028 Study: 418018 PUPS IDs Species: RAT ARccEession S2pec. | Gp SexAge B5B 80g933a53s77e34s23 H1di1et5e/8ees8t76w51 12 12 sreean Grow 12 B8 9o803u3s7n4a2s 1141868s/ess 1133 sseeann Group 13 TR3,ATN0TAL1B4BTOOTAL TRg h7 FBRTRRTs BEFIeREITT eEs EWnSRe ITE gee 3So3s 0Ti8es iu is oioe 1B7H8 80.800 as 80.808 T30888 3poral 8 TRRSESERTA TY gee 8lo8ss 89:.3989 T28oh Reterence Range Bsoo. 33. Ssmoh8ge 1p2ee ApprovedBag--a=l44leote m1eee pate A2ML. 418-018:PAGE H-72 r o-- 0rk BEmAEEEoABF nagen GCg o EiLIfETENTnne :Eo,nd`HBAfRGLUS RESEARCH LABORATORIES, BT8Bh INCDATE SRELEOE SPEC. COLLECTED: w--n-- Sera rT N[ BfL2 iLgEh-Eeem -- .porns 1i33-B, wE[wEo7reexm ff1no::i1e, Preoern ooon wwoorrmm sseo vreen Tfiygefiei fiction h-ot vroorra: ooo ror over Loa Laboxstory Diregstigge: g7a5g9ed Don, aD: R,A sv, saves 418-018:PAGE H-73 INCORPORATED 30121-0168 Fax: 301.977-0433 Client: A5R05GUSSumReEhSYEARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/02/2001 H(O2R1S5H)AM,443P-8A71019044- Date Reported: 02/13/2001 Client No. 1028 Study: 418018 PUPS IDS Species: RAT ANcocuession S3pec. | GpSexAge BBB 000000333335777888232 1111116682223027/632"36 111090 BB 0000333377885 1111683254663289 1100 M5e3an Group 10 CMHGO/LDELS|T 10110185 1178 i11s0.8 GMLGU/CLO.SE 212573830 2i67 253%2,"84 IMDOL/-DDLI|R EMDGL/CDDLIR TRMGI/GDL 332748 33#78 112385538 332 3a1 2i38 333%,4 53056.2 28 06 BBB 000000333535777444789 111111532111579//7528213480 223 BB 00003388778810 111188313/73283176 32 BB 00003388778323 1111833/2*5/328 22 M7e8an Group 2 mi il1o 3 32357 53321% ai311 1235388 i 2109 32148382 i32s72 E33i18 52138238 108 3% 3 EH] 202 5111 8T327523 736%,1 833636 I198.8 BB 00003353775845 1111583368//533374 33 Reference Range 1i4n3 s5s6 - 231170 73149. k5H3 80- 332 234279 820-3 125 Page 1+ 418-018:PAGE H-74 CLYTICS coc sion sons samme vo ar INCORPORATED 301.921.0168 Fax. 301.977:0433 Client: ASB0RU5GTULSDSIHNERGRERSYAEARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/02/2001 H(O2R1S8H)AM4,437BA872019044- Date Reported: 02/13/2001 Client No. 1028 Study: 418018 PUPS LDS Species: RAT AHce:cessionS3pec. | Gp SexAge B5700003358778578 111154430//5533205 33 M5e5a7n Group 3 CHHGOJLOELS|GLMUG/CbOLSE 113338 22384 1145.5 2B62N.3 IMGD/LD-LD.IR 3120 B50e MEGD/LDDLIR 2382 3 32.3 ZMG/mDLic 523710 829%s BB 00003385778598 11113585987/557609 55 BBB 000000333333777866012 111111388666183/77533688873 325 B 0033763 118717" 3 MSe8a7n Group 5 111261221 223882388 23i47 3322s4 22i58s29 ii1ii2ss3 222323730 i2a1 32333 33723818 ETidss 32889%.3 834h.7 832.7 8268067.3 BBB 00000033325777866564 1111118858773647/357785 & BB 00003332776687 1111358843//557850 6& M5ea87n Group 6 111231317 523028377 34i77o 34i07s 211893376 1e6r i2e8s3 3i2 3388 234233 17281.2 2883h 335.2 33%9 28125%.8 Reference Range 5g6- 34s- $0Page 2 + 820-3125 418-018:PAGE H-75 ANI Client: A23R05G2USS3TERREIEESNSECAROBCRRHPEOLARBAOTRAETDORIES3,01-9I2N1C-.0168DBaitteeFaCSx:oEl3A0l1eV-c9Et77eR-d0:43302/02/2001 B(OREGASRBaEso0a- Date Reported: 02/13/2001 Cutent wo. 1028 Study: 418018 PUPS LDS Species: RAT fAececessionS8pec. 3BBeoOemiRmzeEss inHsmeers soeagn Group 7 GpSex Age GHSCOURLEESTST GSLEUjCSEOTSE LMOLT-RDIR HMDRLV-RRDTIR TRREISG. 777 1m H50 o om h5s phii # i3 1I5e8 m2 o 8 HL B8r THHe 815%5 BB8B 8808880033332258777777778323 HH1111ee66ss00iy08/7ee68se00:t88 22& B 8838778 HEE eeagn Group nwJd8is 33007 a3i#s i i3naa nO8#M 3B i i038 2Te BEnhe 03s dBm a a8e 3BBB 8890e9803338832777a0s8s0 HhHmieegltetvyaen/lnesans2 B 8838780 nea 2 seeagn Group 3 ww B0eoo om31s 3233% g $ i sem B 8 am we 3% 8 a Ta BFeNa Waima Bise uisne 3eterence Rang"e s38e. 1- 80- 820- FH: Approved =====fZleeeeo pate --4,00il20 PP! Page 3of 3 4 ANI, 4I8.018PAGE H-76 IS EoRFGRATED sorazioies 600237:2815 oA AUDIT REPORT IhGohoeedrrbeepoortrleiserteedraTbcaentaliocwaeislwaasansdrseowvciiiteahwteeEddPAffrioaGrnwodoicddnoagmLtspealbiowwareenarrcteeeorwryrieetvPphiroearwcttethedeidceFfsOot.Rro mTaenEageement.HP ioncistency and the hTo ee me ethooddosieiuswsciettdhstwhteehreeFdDaAtthaea.ndmeTEthPheAordeGsfooordde,eLsacbtrohierbsaeetdorsytaundPdireasctthiweceerser.epdoornte Gia? fen FGAraAnukdliintor8, Newman seonsor: ARGUS RESE Rl #485 stove: 4/5014 REPORT TYPE: Cn, 73,7, TSH nic 7, PRETH AUDIT DATE: a-150) REPORT TO MANAGEMENT: 2150) AUDIT #: 0-078 Sid3/SGbS/NtITTED , To CLIENT nuk Laman FETS hA aa TTeOT ANTE INCORPORATED 301-921-0168 JR 800-237-2815 atsotspAGE HT) sponsor: ARGUS RESEARCH L465 stop: H/01F REPORT TYPE: cioL AUDIT DATE: a-15-0/ REPORT TO MANAGEMENT: 2-15-01 AUDIT #: 01-121 3/8/01 Ah DATE /SUBMITTED TO CLIENT > 2k Quepnand Vaek Se CEMENT 21570) GLYTICS ogra se st 0 css wo zor B eL sP INCORPORATED 301-921-0168 Bb3att2eFaxSR: Se3Ap01Io-Tr9Et77Re-Ed0433 o1a0j/e1n0//a2c0m0n1 (215) 443-8710 Accession Spec. Gpsex Age IDL-DIR HDL-DIR CHOLEST TRIG 418-018:PAGE H-79 GLYTICS cous sie st 0 amuse wo zr client: ARGUS EIRNeECAORRCHPOLRAABOTREADTORI3E0S1,-92T1-G0.168 DateFax:So3l0l1-s9c7t7e-0a4:33 (215) 443-8710 AJoecsiconession E E "Gpsex hoe TA TIDiopTR a EDL-DIA EE CHOLEST TRwIeG, 418-018:PAGE H-80 CLYTICS INCORPORATED cs res 30121-0188 st Fax: 0s 301-977-0433 wo or Client: AB5R0U5GIULSSDHIENREGERSYEAARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/07/2001 H(O2R1S5)HAM4,43P-8A71019044- Date Reported: 10/10/2001 Client No. 1028 Study: 418018 FO C/S GD21 Species: RAT Recsession S8pec. BBB 000000333777111777132 111000990333;01 BBB 0000003337771i177745 111000889333348 BB 00003377117787 1100383370 M5e8an Group 3 opPsSeexxAAggee 333kEE 333 EEF 33 EF LLDRLUC-RTDRIR o00:89s33 000:::858083 00::7817 T0a7389 HMDeLf-oDwIR cNHeOjLaEwST o0oise33 2lz.ie5sz8: 090:18388%3 l32g3l& 08:88 33188%1 1T8l3s0%s 2R.E5E31 ZMmei/gom 77e.l90y02 7eIllI5eS3s l7e8ss 7.315 BBB 000000333777111728901 111000908444657 44& EEEF BBB 000000333777111888273 111000989348091 4&& EEEF BB 00003377118885 1100893883 44 EFF M5ea87n Group 4 000.:679818 9000:8.36%82 333.330001 9egl.ee9rs1 602:770153 000::3838 3223188 elsenaxn 60:.8383 808%3 33423 lesitd E 3ET]8s 25,033604 71.803%1 Reference Range 0 - 06 - oS - S0Page 3 + 418-018:PAGE H-81 ANI& CLYTICS erosion ss 20 cso vo or Client: ADB SRGUESaRr LIENSECEAORRCo YHPOLRAABOTREADTORIE30S1,-92N1C-0.168 Do ateFaxC:o3g l0l1-e9c7t7a-0d4s33 gots Pesreats assay cttons er wie (215) 443-8710 svuatys 418018 70 /5 coz1 species: TAT PJeEaian allll IS ppFaapunnaMntneT EiIanEEg 1EEi 2fo:: T T 2iiE HE22ie F 32LiEf u3iIn PI PanEa uiRmE f:o@f P=gHg IIgFEE nInFh io4Eb cusp 5 wreoan F HT EEHW TOWaEm oWEa HsEPE ouMammSonmsIiR1sS 6ffx ff otIO W mMgom o ge3ns oao2OooEmnr aoe33mry PP PuiR ImiN S nE ICf:ft MM I o oE ] o3m o3H avon 8 pFoi SBr OETEs guPn Toann Reterence Range - g- g3m - gsve ot 418-018:PAGE H-82 INCORPORATED 301-921-0168 Fax: 301-977-0433 (215) 443-8710 Ft g Accession spec. Gp SexAge LTDLa-DIR a HDL-DIR ChHOeLEST TRIG Awppropvegrovedco ried s e Date --L2H/a@/ka: p---- CAYTICS cvs 500 stro sn chient: N08ESIIEeNECAORREDPOARSOARTNEIDORETS30,1-9r2e1-.0168BttaFaxSS:Se3L0mM1-Ea9E7n7E-S0a433onjenysann o18)M43 8710 Accession ggec. | GpSexAge LOL-bIR HDL-pIR Coieer mma No. II MG/M MG/GM MG/GM MG/GM op3 418-018:PAGE H-84 GLYTICS cose sn, como, wo zor Clients ANGUS RIESNECMONCRHPOLRABAOTRMEIDORIE3S0,1-9I2N1-C0.168DatFeaxG:el30l1s-c97t7e-d0:433 (215) 443-8710 fFoESemsion spec.| Gp sex Age aTR a gpribin ggioonee r zo. B 0036821 11029 1F 0.15 0.41 2.73 8.47 croup 11 ae 0 0 o o 418-018:PAGE H-85 GLYTICS INCORPORATED cass 500 cago vo wor 301-921-0168 Fax: 301.977-0433 Client: AB5R0U5GIULSSDIHRNEGEESHEYAARDCRIHVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/07/2001 H(O2R1S5H)AM4,43-P8A71019044- Date Reported: 10/30/2001 Client No. 1028 Study: 418018 FO LDS REPL Species: RAT Nooc.essionSbip)ec. BBB 000000333666888333358 111111000556751 BB 00003366883378 1111006654 M5ea5n7 Group 13 GpPpSSeexxAAggee LLDRLV-GRD-IR WMDGLj-aDMIR_ CMHeo/LoEnST TRMIo/GoM 13 3 FOFEE 000.::000000 000.::332328 ei3.l!53l20 2112s5.i89z08s norOF 60:0103 00.4475 337398 11s1lled1el T1o0s835 T(32603 31.202983 14058.38765 BBB 000000333666777666335 111000909332331 23FI EEF BB 00003366776678 1100992247 PFI F M5ea8n7 Group 2 000:.091081 000.:338771 434.135319 19s0.ll1s74d6 00::0000 00.:3333 2_3:l8s14 110i17735 "RoEs 3114a T13874736 1101.82864 BBB 000000333666777766098 111000999324891 33FR EFE B 0036771 10942 3 0F M5ea8n7 Group 3 900.:020030 000.1:53a831 233.l37a68 18g6.l10e5929 0:98 0:3: 3167 Tsiss f"0i98e T1a0l3s5t Tl3i2s75 8i.edese RetferenceRanggee 90 - g9- go- 3I Page 3 + 418-018:PAGE H-86 CAYTICS crs ste 0 cam vo or Client:2ABA00RUR2GEEUSSRRIAREREIE"ESNEBCAnORBRCRHsPI0OELsRAsBA-OTREATDORIE3S0,1-9I2N1C-0.168 DBDaaattteeeFaxCSR:aeoL3plA0ol12re-Vtc6E7etR7deS-:d"04:33 0120//0370//22000001 BI Mand Client No. 1028 Study: 418018 Fo LDS REPL Species: RAT RReeecessionSp8ecs orPSsoexxAAwdee ILDRVpaHE~R HMBoLjpoiR GMhoojwoMer_mMmooam BppBoooomnnneaeaerdieteodestss idoiEkFof aF8&l::Co88Is88 o88 SS3eI 2fS.iSi0p5 iH18s.ii3s7 ppoesleeise ieee LooFk 8S:i08 0 8% a 3B 8d seoapn 8T3n 3 a dLaig RmSe Grow 4 BB50oo0n3ae7i7se8 Bocas 1i180n39e85t78 10% 333 FEEE 3k BBeoBaes 18888 33 oF seeean Group 5 2S0a:i.8g082s0 038a.3%4380 nBI3.gEn08 Q9u9.E8s80 8a:888 8e%h 33d Ik T3er TBaeh hueyn enaes B5 ooosnerase 10971 88 FEf Reference Range 08.:8080 80.238 4fi.s8 1m1.k02 80- 89- 89- 8o- rage 4 + CAFYETAISCTS Bcaimmaee sm0a2.ncarsson, wo sr YHEaLP 905 BRI cxporseocn (215) 443-8710 NAocecession S1Bpec. | Gp SexAge LDMLG-/DMI|R HDMLG-/DGIR CMHGO/LGEMST_TRMGI/GGM 418-018:PAGE H-88 CAYTICS cussion sie 00 conse vo mom Client: J3BR5EE5GSH0USE8SH,oCRErIEeESNSEACA,PORRCRHC"PToOLRnABAaOTREATDORI3E0S1,-92T1N-C0.168BDfaa3tt8esFaxSRG:oeRlp30loA1sr-Ect97teS7ed-Yd:0s43301a0//03r0//220000m1 Client wo(.2151)028443-8710 Study: 418018 FO 105 REPL Species: RAF Ffeolioensgee Re aRa aGn Gweaa ae T yen croup 8 WOH ap ham axoup 5 Reference Range g- gg gspprovedEora gs renee pate RUE 418-018:PAGE H-89 Client: ABRGUES RSIENSECEAORRCTHPOLRAABOTREADTORIE3S0,1-9I2N1-C0.168 BDaatteeFax:SCoL3l0El1-eE9c78t7e-"0d4:33 3/07/2001 BTEEE)EMRidSo t,hal ooue- Date RePpSorted: 10/10/3012001 Client No. 1028 study: 410018 FO TDS REP? Species: RAT Rreosseneien g8pee a Be nee RRUET IRATRRR GgOioRus Bmea, 3Fpogooaomseecieeseesos audHisazleeerr Bosteser dist T111 Eo2f 1 gGgGe0eae0e gig soeoeerys eh bbLbsnals bE oa agisd gn BpBoogoogosseeeseeccsrsss zHeuaseglees 1I11 ofkfof 8 gaesos dizer 1b ggggeieie gee8eee%rse nLBTeigEl eg BEeE gre eee ms BES BpBsooeommmmeeeeesseniinsozidiitinlmynl 1111 p}ooff eggg::ee8800 &gg8i%oeeas IILLSLnES g8ogdat5 sreeann 5 0Th iugineian wor 3 poBpoooosssneeeemssnz 1nnnseeezr 111108 pEEf Boments en 18 0gge.::0e880 ooagieni 1nLr8dBE9e 88ap.SR2d0 ge oy 1S ee 3poamenntge endet 1B8 F gees gie ELRm iodF Reterence Range o- o- g- g- ng 0 0 0 0 Page 14 SSOISPAGE R90 GLYTICS INCORPORATED HJEERREESETo.us (215) 443-8710 oc senst20 csr wo wor 301-921-0168 3238 SUT Fax: 301-977-0433 cosorsmocn pete mereEten Topi AJaecaiacn ession 3Spec. | Gp Sex Age IDBLi-aDIR a HDL-DIR CgHoOnLEeST T peRIG 418-018:PAGE H-91 PEs, Bape INCORPORATED Btu 301-921-0168 B58 SSLSET Fax: 301-977-0433 0/07/00 AJosce iconessionSp ec. Gp Sex Age Ba IDL-pIR a HDL-DIR oe CHOLEST TRIG TTT BOISPAGEHS Clients agus gIeNsECAONRCHPOLRAABOTREADTORIE3S0,1- He. 921-0168 Date.Fax:Go3l0l1-s9c7t7a-0d4:33 (215) 443-8710 AecScession S2pBec. Gp?Sex Ag9e LWDJLM-D.IR HMDOLJ-GDMI.R_CMHGOILGEHST. TRMGI/GGM 9 0 0 0 0 PTS---- INCORPORATED Ly (215) 443-8710 301-921-0168 Fax: 301-977-0433 Sass neporeess ori0s20m jJociceession spec. Gp SexAge Ta ILDL-DIR pa HDL-DIR SE CHOLEST TRIG J 0 0 0 0 418-018:PAGE H-94 CLINYCOTRPIORACTESD socimssion 301.921.0168 sue Fax: 0 sansa 301-977-0433 vo or Client: AB5R0OG8PULSOSHIERNEGERSYEAARDCRHIVELABORATORIES, INC. HORSHAM, PA 19044- BDaattee RCeoclelievcetded::TM 02/07/2001 Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 Study: 418018 FO LDS REP2 Species: RAT RHoe:cession S8pec.| BBB 0000003336e86e84d5ss0 111111533535234 BBB 00000033388866e33s3;t 11111833888785 M5ean Group 4 GpSex Age IWBEJLGHD'"IR 444 EEEF 880.::00830 iii 000:::000606 T0o3o8s TMGIj-obMT~R_cMhGojLGgHer TMRGI/GG 0o01sia 331.9g075 1i130oi.g2ls2 00O30338 _ l1ilo9ar3r Fi1i31lllag8zl3 010%s 2H:0e4 815h,3E38 BBB 0o0000j33666e3s334 111111585556881 555 EF E BB 000000333666e83337s5 1111ii3ss8ee2sd8 355 EEEF BBB 0000003338c88e86e0;l 111111333666s7s 555 E BB 0O00338g8e6e4s 1111537710 35 EFF M5e8an Group 5 000:.:000800 009.:110718 1il.ea7ss4 6sg.il3za8ez 008:::000000 0:00 080:01d83 8:33 iiigedsse 1g 7lslid3soe 8i37 600:::000000 0oo0oi% iiildliess 8esill5eo5ss 6:00 0:07 138 3ise 9s Tloozs ltiesfstz s1.47288% Reference Range 9$ - 9 - 80- 96Page 6 + 418-018:PAGE H-95 Cltent: ARGUS, RIENSECAORRCPHOLRAABOTREADTGRIE3S0,1- Tie. 921-0168 Date Coliscteds Fax:301-977-0433 (215) 443-8710 AFictecession gSpec. op Sex Age IiDLa-DIR a HDL-DIR CHgOoLnEeST TRgIuGe TT 418-018:PAGE H-96 Chtents Agus gIaNsECAORRCHPOLRAABOTREADTORIE3S0,1-9T21C-0.168 DateFax:So3l0l1-e9c7t7e-0d4:33 (215) 443-8710 jAe cmaiconession spec. "Gp SexAge IBDLT-DIR a HDL-DIR CRHOLEEST TmReI,G TTT ae 0 0 0 o 413-018PAGE H.97 CIYTICS orcs 0 20 casero wo mom Client: A3H5Ri80G8USBERIRENSRECAORBRCRHPIOELRAABOTREADTORIE3S0,1-5I2N1-C0.168 DBaatEeSFax:Co33l05l1-e29c78t7e-90d4:33 02/07/2001 BTOORRESAMatBha ab10044 Date Reported: 10/10/2001 Client No. 1028 Study: 418018 FO LDS REP2 Species: RAT Ahececession S9gec. 5pBBoooodsimdeersseees HLnHieeeerrn ppBoeodngeaem Bona Htneeeles te Bee ue seegan Group 9 Gpsex Age LHDUL-RDTIR HMDL-RDIR CSHOELTEST TRRIGG 33323 k:EoFf g08ao8%es eooennHBl hn B2.0ER8 u18n w1.Hi98 585:k& 5E 8a&:888 a8 hoemHB en bI nei nh ieaEs uf 3 of 88% Sb nn 83 BEri Thin s3a3o% Reterence Range ae 0- o- 9- o- 0 0 0 0 TTT ApprovedSagn e 5 oe f pate ---2/22/el 418-018:PAGE H-98 GLYTICS os syeon ssn am, crs, wo osm Cltents EPEVL ESBIEENICOGREEPOLRAARTTERDISS3.01-9T2H1e-.0168RD2a5tF8eaxSE:o3vE0l1e-RE97tS7e-s0:433o2j0m/20m BB ANooscession S7pBec. | Gp SexAg9e I[LJDLo-nDIR HDReLj-eDmIR CMHoOjLeE.ST TmRoIcGk J 0 0 o AISOISPAGE H99 GIYTICS coms sev sun20 crus vo asm Cotont: aNocvtsSaMIRpSNaECAROCRH.POLRABAOTREATDORI tN E3S0,1 THC. -921-016 8 aBt5s8FaxSS:o3lL0l1g-Se97t7Ra-d04:33 sate Reported: 1002//300//2200001 (215) 443-8710 jazeoeioen saionBesser7Ssee Reheee IgBiEpER EWian cSoEmTs gWEma ; 9 | 0 0 0 0 418-018:PAGE H-100 Clients AYRGUSEAIESLEEAGRCPH dARADaGtRtATsORIES0, STei.estBBoan2ts2.SnSSelilSnsSctaEatdh:er02/07/20m EEE cone Se operon toysoyors (215) 443-8710 ANcocsession S5pec. | Gpo Sex Ag9e LMDGL/-MD.IR HMDGL/-GDMIR CHMOGL/GEMST TMRGI/GGM 418-018:PAGE H-101 GLINYCOTRPIORACTESD 3c01i-9e21s-01608 0 Fsaxm: 0301-8a77-0n433e vo 2m Client: 2A5R8GUSSuBREERSYEARDCiHivELABORATORIES, INC. BDaattee RCeoclelfevcetde:d': 02/07/2001 H(O3R1S8H)AMH,43%BA8m1010044 Date Reported: 10/30/2001 Client No. 1028 Study: 418018 FL C/S GD21 Species: RAT wAoc:cession S7pBec. | GpSexAg9e BBB 00000033366639339830) 111000983588850 323 BBB 000000333666933939335 11180083366]32 333 BB 000033663357 110633is 33 MBeaHn Group 5 LMDGjLG-HDTMIR 000.::000008 o00::l00e00 06::0000 s9 EWDOLf-wDIR CMHoojLoEMST TRWIo/Gem og0oo0drs 333.8A783 575.:13383 0og0eo8s 3l3ed%rr 85318273 goaod 3383s 35:782 BTBa Tuies 5B.1B33 BBB 000000333667930908088 111800395777425 & & BBB o0000o333777000000338 111000339778880 & BB 00003377000035 1100%38821 & N5e%an Grow 6 080.::000080 03o.l0e0ds3 44i4ggs 737.19g398 00g::e00e00 3gol0eosdr 4if1lE8o 193313%s g0r:e0e0 o8tedd 3agd gflii $? Toafs TWaigtes L7.E7S88 Reference Ranngge $9- 96 - 89- $I Page 4 + 18.018 PAGE H102 CAYTICS css sn 0, unr wo Client: A2BB55RovOsGa,EUgSBRuSnREEIESNEEBCyAORCCRH1PHoOL0ARuBaAO-TRAETDORIES3,01-9I2N1C-.0168DBDaaatttFeeeaxRCS:eoS3pl0oLl1re-St9c7eEt7de-A:d04:331002//3007//22000011 Client vo(.2151)028443-8710 Study: 416018 F1 C/S GD21 Species: RAT fP ee fi RIGHT r RII gout ie, BBBB3 g0gggo0ooaa3a3ut7tt7ooo00na000ess63 11i1100o002s9o88ee88srs85 77JJ7 BBooggawwuereeitlzo 1i1sg0ts0 777 Mean Group 7 0Fggg.:Eee08I00 0B8 .ge0igs5e 5NsI.0GtR0 8eae.l8Rne8 - s5g:ee8 8 88% NNNEyE Siss yh 38 8T3i agss LTams Reterence Range 8o- 89- 89- 8a- , ApproveSag--a o5wASmzeeoeeemen pace 2/30/01 418-018:PAGEH-103 cident: JREUS BIRNACROGREPOLRAARTIEIDORIES30,1- He.921-0168 pate Soligctar: Fax: 301-977-0433 (215) 443-8710 Ajacecaicon ession 3Spec. GpsexAge LpL-bIR HlDlL-lDIR EN CHOLEST TRIG TTT 418-018:PAGE H-104 CCLYTICS cs ress 0, ct, wo 2m INCORPORATED 3015210168 Fax: 301-977-0433 Client: A9R05GUSSHEREEHSYEARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/07/2001 HB(OU2R1PS3L)HDAIMN4,G43BA-A871019044- Date Reported: 10/10/2001 Client No. 1028 Study: 418018 F1 LDS REPL Species: RAT RAecscession Sip)ec. 5BB000000333777111333890 11111100022357 BB 00003377115441 1111003280 MSe8a0n Group 10 Gp SexAg9e 111003 EEEF 1100 FF LWDRLJa-hD'"IR HDMGLj-oDmIR CNHOLGEST 080.:1000000 T0Oo1di0yl8 s4dae8ze 001:0008 00l%od 33388 06 1To8729 a3 z TmRoIjGo i11g28.1ia5d373 110o:l3i8o i1l35.1538 BBB 000000333777000777234 111171000221319 111000 MM BBB 00000033377700077757 111111000222456 111000 MX BBB 00000033377700077198 111111000323807 111000 MM e5a5n7 Group 10 000::.000000 0o01.f00f88 a4a.il2li2s 11T23d.1l93s71s 000::000000 0000l%os7 ii3l3El9s 11383i13a28y3 880:::000000 &00i03k7% 333183a88 8g_is%gis7 06 T1o6s3e3 TWizrs3 e11s.81 Reference Range 80- 80- sI 0S Page 2 + 418-018:PAGE H-105 CLYTICS INCORPORATED os ins se 0 sam vo om 301.5210168 Fax: 301-977-0433 Client: A9R05GUSSHEREEHSYEARDCRHIVELABORATORIES, INC. DDaattee CRoelcleeicvteedd:: 02/07/2001 HORSHAM, PA 19044- Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 Study: 418018 F1 LDS REPL Species: RAT HAo:cceSs8pesc.i|oGnpSexAgeLRMGL/DGMI|R EMDGL/-GDHI|R CMHG/oGLM_E__sT BBB 000000333777111444423 111111000233921 oniI FFE 0001:000000 0o0.l:0e8%s7 3ai.lil72d86 BBB 00000033377711144475 111131000333348 iIoI EFEF 000:::000000 090801:0sa8 33all87o7ss BBB 000000333777111445890 111i11o00l33e87 111111 FEEF 600::000000 800:0l9os88 3a3l:lg4sa8E M5ea87n 60 Tiotss 3T.i9s3 Group 11 TMRGI/GGHM 11e9711l53s22e g58i:1s15t03 193a1i04d83e 41.18152 BBB 000000333777000888013 111111000323291 11o11o X BBB 000000333777000888345 111111000333335 o1o1 Hu BBB 000000333777000888878 111111000333678 111111 XM $Me8a7n Group 11 000.::000000 0oo.ll1eo1ss 3l4.34s75 26S0.s07e81 000::000000 0001ll80o88e 4&4l.4l33i1 1708.1l57171y 8081::000000 _ o00:o00r33 l43li:ga8s i8si1is5sz% o 1T0o3s77 KTai;]021 3105.63416 Reference Range 0 - 0 - I S 00 Page 3 + 418-018:PAGE H-106 INCORPORATED 301-921-0168 Fax: 301.977-0433 Client: AB5R0UG5TULSDSIHNERGEERSYEAARDCRHIVELABORATORIES, INC. HORSHAM, BA 19044- BDaattee RCeoclelievcetde:d: 02/07/2001 Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 Study: 418018 F1 LDS REPL Species: RAT RNeescessions3pec.| GpsexAge THOOL/-GDHT|R EMDGL/.GDHI|R GHMoGL/EGST TRMGI/GGM BB 00003377118521 1111004851 12 oEF 60:.0000 00:.9039 34.1133 1122.0941 M5e8an 0$ alo12s 3l,ag7s8 1e2i.s475 Group 12 B 0037083 11049 12 ou M5e8a0n Group 12 0.00 0.07 _ 3.87 _ 8.82 0 8"07 33757 6Tslez B 0037153 11059 13 F M5e8an Group 13 0.00 0.07 _ 4.39 11.96 o 8"07 io380 0Tiilse B 0037090 11059 FEY e78an Group 13 0.00 0.04 3.31 _ 8.45 s0 8"oa 38517 8gas Reference Range 0S - 0 - 0 - 0- Page 4 + 418-018:PAGE H-107 CCLYTICS soci see. st 0 css wo rr Clients AyReGsUS,SReIENsSECAORRCHPOARRAORTAETDORIES30,1-9T21G-.0168 she 3G3a6t2eFax:SGeS3b0Ii1Ss-9cE7i7Ra-Y0d4:33 eve Severe 0a2n/r0s2e0r0e (215) 443-8710 Accession pec. Tp Sex Age IDL-DIR DL-DIR CHOLEST TRIG No. I MG/GM MG/GM MG/GM MG/GM B 0037097 10921 2F 0.00 0.05 4.26 22.52 - ae 0 0 0 o 418-018:PAGEH-108 CAINYCOTRPIORACTESD 301-c620r-016s8ve sFatx 30001-9c77a-0n03o3n=wo ss Client: AARSGUSEEREESEARBCRHIELABORATORIES, INC. BDaaEtSe RCRoAlAlIeScEtSeYd": 02/07/2000 BSRERANS Ba 1004s- Daatte e RepoRreptoertde:d: 10/10/2 (215) 443-8710 P 30/200 Client No. 1028 Study: 418018 F1 LDS REPL Species: RAT jfcecesaion g9gec. | Gp sexAge B 5B0o0o3aR7r0e1iI 0s 1eE 0s0s3e5 333 H Keepan Group 3 DWDRLpTIR mMDRLFoEbRiR gRroEtSeTsr 808:.:800808 808:.:800158 i 53.:083 78 TSoEa LBEn zWmEioG 2848.:%788 V1eadm ppB 0ao03gs71es05 ii109es57 e:52 E FpBRo0Ed0E3i3sH0t8E1iH8d0e6Ie8 32 & Neepan croup 5 08a.8080 eg 088.0%%8 8% 3ni.dY95 1316.8866 fi ih a8:888 88:%88 n$h8 538d 58 T0o8g T48i8o 19gdEa0 pp8 0oo03d7e0e43 I110n9ss5e7l B 003%ie 1806s 3352 M 30837855 1886s 3 Reference Rang9e aa0a.88080 8o0.:n088 Sey 4E4.3E51 3 819.%77 3 ID 88 8% ne le 80- 8o- 8o- 80- rage 6+ 418-018:PAGE H-109 Client: AB2RSG8US2BMREIENRSECAOBRRCRHPIOELRAABOTREADTORIE3S0,1-9I2N1-C0.168 BDaatteeFaxR:CRo3lE0l1Ee-V9cE7t7Se-Y0d4:33 02/07/20m BB ERRAM Ba d1s0ss- bate Reported: 10/10/2001 Cuient No. 1028 Study: 418018 F1 LDS REP Species: RAT ARceceession S8pec.| 5 ooares 10970 NBeEaRn Group 5 pBBoOom7Mpia 109 IE HBeEan Grow BBoOo37mS01e0Se10E0n77 segaen Group 6 Gpsex Age 5H 8ffEF 68& LHpeyLaoRrR jpMRLyoaRnTMiR ghSoErEsRsr mEEmjGck ooo o.o8 3.01 27.48 38 T8o8s huaBms zinnaer 808:.:808088 808.&0%87 I2n.if68 G18 2.n43 38 Te aheEs 125840 080::.880080 808.077 333.%80 17.37 Re 83 BTie gWPen 31Hm ReferenceRanngge eg - 9a - 8- @- rage 7+ 413-018:PAGE H-110 GLYTICS so crass ss 0 canoe vo or INCORPORATED 301621-0168 Fax: 301.977-0433 Client: 5BA0RUSGTULSBSIHNERGEEHSYEAADRRCIHVELABORATORIES, INC. DDaattee RCoelcleeicvteedd:: 02/07/2001 H(O2R1S5H)AM4,43-PA8_71019044- Date Reported: 10/10/2001 Client No. 1028 Study: 418018 F1 LDS REP1 Species: RAT RNoewcension S1pee. 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A0lda 418-018:PAGE H-113 LYTICS imsee 0 20 csr wo srs Chtent: AL ngus L sIENsECNOP RRCHPOKRAAOTRAETDORIR3S01,-92T1H-C0.168 DatFeaxE :Go3l0l1s-L9c7t7a-0d4:33 ETRLEE, ere wpe. Serer ction ve. 1078 study: 418018 F1 108 REP? Species: TAF Ey Rocessiongpec. | Op sexAge a EpLbIn EE morph a ghormer gue. ae 0 0 0 0 418-018:PAGEH-114 CGAYTICS cre sve site cams vo msn rtm JeIEnLE LPT ov INCORPORATED 301-921-0168 oSErFIaxC: E30L1-L97E7-D0:433, HORSHAM, PA_ 19044- (215) 443-8710 Date Reported: 10/10/2001 Ey hccession spec. "TT"GpSexAge Ta IDL-DIR HDTLa-eDIR ue CHOLEST a TRIG ! Group 1 418-018:PAGE H-115 (0/INC0OR)PO(RAOTESDJ301-621-0168 ---- Fax: 301-977-0433 Client: ABSR0OG5TULSDSIHNREGEESHYEAARDCRHIVELABORATORIES, INC. RORSHAM, PA 19044- DDaattee RCeoclelievcetde:d: 02/08/2001 Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 Study: 418018 F1 LDS REP? Species: RAT RRoe.cession Spec. 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"Gp SexAge a IDL-DIR HoDL-eDIR CoHOeLEST a TRIG aroup 32 A J o 0 0 0 413-013:PAGE H-118 erD4)(OSJ INCORPORATED 301921-0168 ---- Fax: 301.977.0433 Client: BA9RU05GZULSBSIHNREGEEHSYEAARDCRHIVLEABORATORIES, INC. DDaattee RCeoclelievcetde:d: 02/08/2001 HORSHAM, PA 19044- Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 Study: 418018 F1 LDS REP2 Species: RAT RNoe.cension 5pec. 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B 0037339 11515 2 0M 0.00 0.12 3.28 13.01 - 418-018:PAGE H-120 cLYTICS 200 Gira Sree, Suite 200, Gaithersburg, MD 20877 Client: AARVSGUSERIRENSECAORORCRHPOELRAABOTREADTORIE3S0,1-9I2N1-C0.168 BBaatteeFax:SCoe3l0lM1e-C9c7tE7e-S0d4:33 02/08/2000 E(5S15H) E4R43B-a87101004s - Daattee ReRpe>oprotretedd:: 10t/1a0t/2001 Client No. 1028 Study: 416018 FL LDS REP2 Species: RAT fReeaseGbnieae ioTno se hee HRpAn Ma ya GSEOIGERST mHeen FBpBoooRnmgEasa1nc1ene5s3t 333 B Boome al 3 eeT0::eo08es88 BeoooIiisp 333h.g%%35 1iW37.Es12 &88 oi Rf BE B[ReICpHpAe 3 x &&::8588 8ofp 3 3 al3s sdeepan TS I a Toe aueis 1IR7Ba Group 3 p5ppooooooyanreneescise eie1s5ese8 3325 OEE pops ine 2 aa0S.:e080p08 a0 aiooRdst Se R44A.E%02 2agW.3EeE2 BE WE p[p pRoroooeyruoiedelraieeeetBA 333 kEF poaaiee 3 F e08::80888 ES 8g%eaTEHNEaIYe g:08 80 3p ai0e ahpgg Bee Hs 3 ok eeagn e585 Si i 38 7 7 an sNeEr AaEw Grow 5 Reference Range 80- o8 - 8o- 8gPage 8 + 418-018:PAGE H-121 GLYTICS cso ss 2m. soso, wo sr Clients AERGUELmIeNEsTCAORRYCEPOaRBACRTAETDORIF3S0,1 The. -921-0168 aBt3Fe2axSG:o3H0l1iI-v9e7Tt7-sE0d4:33 oaj0nr20m (215) 443-8710 AJecssciessiongggec. GpSex Age Ea IDL-DIR a HDL-DIR CHaOmLeEST a TRIG 413018PACE H-122 CAXTICS sesso son 0 samen om INCORPORATED 501521068 Fax 301.977.0433 Client: AARBSGUSSRREESYEARDCRHIVLEABORATORIES, INC. BDaattee RCoAlTlVecEtEeYd: 02/08/2001 HORERAN. Pa 10044 Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 : Study: 418018 F1 LDS REP? Species: RAT Rfeeceession s9pac. | GpSexAge BpppeoeounNompplmTALinnngeeeTeeEe 88 Kou om BBpe0on0tp3e7E pep sLHseEese: EE eoapne Group 6 GHpTLHbiR mMRryeRpT 20a8.:80880 Sd0a1iri2 0.00 0.08 38 I HB GSrEonTer mWiEo.G 32n.7b88 2% 18aE.s7s6 iE 3.7 9.75 Suaam saeEn 5ppooeo3s7saess sHneEe 777 OEfFF ewagn Group 7 08-.:0808 oiubi 44-2 1B9.a32 83 o BE hfy E173E50s BpBooeomzmpe mnmaesSz 777Mok Moepaen Group 7 0.00 _0a1 3.25 _ 8.48 38 8TT 35s 8hae Referencee RanRangee 09 - 9o- g0- 0- Page 10 + 413-018:PAGE H-123 AINYCOTRPOIRACTESD 230001G.i5r2a1r0d1S6t8eet, SFuaitxe 230001..9G7a7t.hr0s4b3u3rg, MD 20877 Client: FA3RSaGUtSShRRiEESYEARDCRHIVELABORATORIES, INC. BOREAS Ba 10044- DBaattee C8o3l2l9s9c6t8eYd": 02/08/2001 Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 Study: 418018 FL LDS REP? Species: RAT FfooscessionSBpe=c. Gp? Sex Age LMDGL/-GDHIR HDMGL/-GDMIR_ CMHGO/LGEMS=T TRMGI/GGM 5ppooooi3d7saeses pois 1mL6e0e:0 HES 8&& EfFF & 3&o::o88o88 8:88 8e0:09n88 di 3I3.:h688 a A1L1N.Ei2s4 - BBBoooaiardsss B o03iss nhheeesses ess 888 & 8ae8::8888808ei84nf8 d4adb%st fl La5hwiiiesd BS BBoOoSsSe HEedE: 88 OF a8:888 d8k% 2 Ae aHdii sBeEaXn 8To8or a ios 3W7Pa disug0ses Group 8 p8p0oo03dp73ee82 pone nn1H1eEe60:Ee0 855 Mom pBBoaaduses pons HHeeet:ee Les 8o8&s BBoERImNneG: 88 Kgeoapn Group 8 gg&0:::.08008880 S80d9.pb81 f3E2.3eE885 15.00 am8a 88 &e::88e88 8 8e&%B E 3n3:Bf B aH8is 8e:g8 88s% I3sE ai8gE 38 TBeEi 5N5Ws E1,d3 Reference Ranngge 08o- 08 - 80- o8 Page 11 + 418-018:PAGE H-124 CCIYTICS csi sn 30 mrs wo mr INCORPORATED 301-921-0168 Fax: 301-977-0833 Client: A5R0GSUSSHEREEHSYEARDCRHIVELABORATORIES, INC. DDaattee RCeoclelievcetedd:: 02/08/2001 BHOURTSLHDAIMN,G PAA 19044- Date Reported: 10/10/2001 (215) 443-8710 Client No. 1028 Study: 418018 F1 LDS REP2 Species: RAT ARecscession S8pec. | BBB 000000333777445990980 111111666111321 BBB 000000333777358000312 1111l16e61l14e5 BBB 000000333777383000546 111111666111875 MSe7a8n7 Group 9 GpP SSeexxAAggee s59 rEEF 989 FEEFF 9SFFF ILDRLV-GHDTMIR 0o0ll0oo00o" 000ll0o0000 000ll00000 0 HMDeL-iDoIR o00.ll11d65 o00llll11oz0 _ooo0rllloo0s 1l1o8o cHMoiLGEST l33.ol9ss0 3339lie2378 3337l8e20 3",373421 TwRoI/Gow 1113331.l88g722 1F007l1i09918 2a13i11ll1o93y0 15750.36511 BBB 000000333777333999323 11i11l666111312 999 MMMHH BBB 000000333777333999756 11111166e11l45e s99 uMM BBB 008000333777333990890 1111116e61i1785 995 MMmHM Y$e5a7n Group 9 00.0000 000 00.:1100 012 32.:1876 176..1641 368 23.50 008:000000 00oi001d33 33l:15e81 i1128al7el72 00811.000000 00o.:u01i83 323l1l270879 12079i.205780 o T15i3o6 3J.a2y08 715i.s4s38 Re2ference RangRange g9 - 39- 03 - 3o0- ApPErpOYrovedagS e 12 of R 12 9 4aad 418.018:PAGEH-125 CGLYTICS wosumnmoassaimno INCORPORATED 301-821-0168 800-237-2815 QAAUDITREPORT GTThhoeeodrfLeiapnboaroltrartleoiprsoytretPdraabncedtliocawelslwaasansdsroewcviiitaehtweeEddPAfroaGrwoocddoamLtpaalbiowarenarcteeorwryietvPhireawctethdieceFfsDo.Ar Amacncaugreamceynt.and consistency and the findings were reported to TSfhcneeucromametptelhlioyidnscreefulwseiectdths twhteehreeFdDaAtthaea.ndmeETthPheAordeGsfooorded,eLsacbtrohierbsaeetdorsytaundPdireasctthiweceerser.edpoomret Fra2n.k2lin eB.aNrew,nan QA Auditor sponsor: __Afcys 485 REPORT TYPE Clb, 04, TIC AUDIT DATE 9-3-0] sTopY: _4{)50if REPORT TO MANAGEMENT 10-32-97. AUDIT # _9l-9ob Safe/s.`fsorumm7Tr0TEDBRTR . 5MocDkIRECTOR RNAGEMENT 73010] APPENDIX 1 BIOANALYTICAL REPORTS (PRIMEDICA WORCESTER, SPONSOR AND SRI) 418-018:PAGE 1-1 FINAL REPORT VALIDATION OF A GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC METHOD FOR THE ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR PRIMEDICA ARGUS STUDY 418-018 Primedica-Worcester Project Number: PABX-BVA PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 418-018:PAGE 1-2 r PRIMEDICA FINAL REPORT VALIDATION OF A GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC METHOD FOR THE ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR PRIMEDICA ARGUS STUDY 418-018 Primedica-Worcester Project Number: PABX-BVA Submitted to: PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 Submited by: Pr5i7 mUniionWoSrceeester `Worcester, MA 01608 Primedica-Worcester Report # PABX-BVA-01-62 Page 1 of 184 Issue Date: November 7, 2001 WD iadk 2,b LL pl woolt Mark A. Netsch / Date Project Scientist Departmentof Analytical Chemistry Primedica-Worcester 418-018:PAGE 1-3 OPRIVEDICA Primedica-WorcesterProject number: PABX-BVA Page2 Final Report For Primedica Argus Study Number: 418-018 COMPLIANCE STATEMENT This project was conducted in compliance with the Good Laboratory Practice Regulations, 21CFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision/ recommendation on compliance with principles of good laboratory practice. Official Journal of European Communities: Legislation 32 (No.L 315; 28 October): 1-17. There were no deviations from the aforementioned standards, which affected the quality or integrityofthe project or the interpretationof the results in this report. EO. Qutb wtrfo Mark A. Netsch /Date ProjectScientist 418-018:PAGE 1-4 GPRIMEDICA PrFiimnaeldiRceap-oWrotrc_e_s_te_r Project number: PABX-BVA For Primedica Argus Study Number: 41P3a-g0e138 QUALITY ASSURANCE STATEMENT `The following are the inspection dates and report datesof QAU audit/inspections for Validation Of A Gas Chromatographic-Mass Spectrometric Method For The Analysis Of Mevalonic Acid In EDTA Rat Plasma For Primedica Argus Study 418-018, Project `Number PABX-BVA. Critical Phases Date Inspected Date Report Submitted to Project Scientist ~~ Management 1. Laboratory Procedures 10/19/2000 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 ofA Gas Chromatographic-Mass Spectrometric Method For The Analysis Of Mevalonic Acid In EDTA Rat Plasma For Primedica Argus Study 418-018, report number PABX-BVA-01-62, was reviewed for compliance with the Good Laboratory Practice Regulations, 21CFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision/ recommendation on compliance with principles of good laboratory practice. Official Journal of European Communities: Legislation 32 (No.L 315; 28 October): 1-17, on 11/7/01. The results as presented accurately reflect the raw data. Nu av A . Quality Assurance Auditor wos Date 418018:PAGE LS SPRIMEDICA FPirniamledRiecpao-rWtorcester Project number: PABX-BVA ForPrimedicaArgus Study Number: 41P8a-g0e18d TABLE OF CONTENTS PageNo. COMPLIANCE STATEMENT......c.cooomsmsmmssesssmssssssmsssssssssssssssssssssssssssssssssssssssssssssssss 2. QUALITY ASSURANCE STATEMENT .....ccoooonmmmmmmmrmmmmmsssssmsssssssmssssssssssssesssssssseses 3. LIST OF TABLES AND APPENDICES......cocooeconmmininsmsmmssssssmissssssmssssessssssssssssssssoses 3. CONTRIBUTING PERSONNEL wc... ANALYTICAL REFERENCE STANDARD CHARACTERIZATION/STABILITY ..... 8 ABSTRACT... 10 DOTRODUCHI mmm 1 EXPERIMENTAL DESIGN ......ooovmsvnnnimninsnssssssssssssssssssssmisssssssssssssssssssssssssesssssssss 11 Materials and Methods ......... essstsssemsssssssesmsss seems 11 VAlGBOD DESIGN... 11 Validation DESOHPHON courenscrmsomenmmesmemesmsmsensensmmmnssesseses 11 RESULTS AND DISCUSSION....cccouuurmummmmmmmsmmessssessssssssssssssssssssssssssssssssssss 14 Validation RESUNS............cooovvsmirrrsssmisssssssssssssssmisssssssmsssssssssssssssssssssssssssssssssssssss 14 `Water/Plasma Extraction Comparison. ................ ssssestrrssenrsesssensremerers essen 1 oT YT ERE spun pmm------16 SUbIIIY rrr mm-------- 02 BEES1 (0)6 SS S------------------ 418-018:PAGE 16 PRIMEDICA Primedica-Worcester Project number: PABX-BVA. Pages Final Report For Primedica Argus Study Number: 418-018 LIST OF TABLES, FIGURES AND APPENDICES TEXT TABLE 1 Validation COUTSE uo. PageNo. 12 FIGURE 1 Day 1 Validation Calibration CUIVE...........cccevuveeesssssssesssssssssssrssssssssesssss2s0n TABLE 1 SummaryofInterpolated Mevalonic Acid Lactone QC Standard Concentrations with Inter and Intra-Day SUMMATY SAHSHCS......cvvewrsrssssmmssssssnsssen 2d TABLE 2 `SummaryofBack-Calculated Mevalonic Acid Lactone Calibration Standard TABLE 3 Mevalonic Acid Lactone Extraction Data & Recovery Calculations .................25. TABLE 4 Internal Standard Extraction Data & Recovery Calculations .............ccocuueererern 26. TABLES `SummaryofLeast-Squares Linear Regression Constants and Analysis Dates ...27 TABLE 6 `SummaryofQC Standard 72-Hour Room Temperature Stability in Matrix........28 TABLE 7 `Summaryof QC Standard Freeze/Thaw (3 Cycles) Stability in Matrix...............29 TABLE 8 Summary of Interpolated Mevalonic Acid Lactone Dilution QC Concentrations for Dilution Validation............cceeceeeveeususne ener snes 30 418-018PAGE LT GPRIMEDICA PFirniamledRiecpao-rWtorcester Project number: PABX-BVAForPrimedicaArgus Study Number: 41P8a.g0e186 LIST OF TABLES, FIGURES AND APPENDICES - CONCLUDED PageNo. TABLE 9 6-Day Room Temperature Final Extract Stability Back-Calculated Calibration Standard CONCENMTALONS ..........vvoreuvrrrsrssasssrrsssssssssssssssssssssssssssssssssssssssssssssees 3 TABLE 10 6-Day Room Temperature Final Extract Stability Quality Control Standard TABLE 11 SummaryofControl Blank EDTA Rat Plasma Concentrations............c.ceeeree:33 TABLE 12 Summary of Unique Lots of EDTA Plasma CONCentrations ............cccuuevsees3e4n TABLE`S1u3mmaryof Internal Sandard Peak ATEAs..........uvuosrssonnnnnen35 TABLE 14 S`SuummmmaarryyoSftPatliastsimcsa QC Stand4ard Concer ntrationn s with Is nter- and Intra-Day31 APPENDIX A LADOTRIOLY MEMHOA.....corrrsssrceerrrrsreersssssssssssssessssssessessssssssesssssssessssssessessssn 39 APPENDIX B Representative Chromatograms from Validation Day 1 ..........cccwummrrrsinnnen5n2. 418-018:PAGE 18 GPRIMEDICA Primedica-Worceser Project number: PABX-BVA Page? Final Report For Primedica Argus Study Number: 418.018 CONTRIBUTING PERSONNEL PROJCCt SCC vrs: Mark A. Netsch, B.A. RESEAICh ASSOCIAE woven. Heidi Gagnon, B.S. Director, AGIYtical CHEMISTY... JES A. Jersey, PRD. REPO COOTINALON .cocrvvvrrerssssssrsssssssssssssssssssssssssisssssscsesrssBesTsEsNeAeeAs L. Brooks: 418-018:PAGE LY PRIMEDICA Primedica-Worcester Project number: PABX-BVA Pages Final Report For Primedica Argus Study Number: 418-018 ANALYTICAL REFERENCE STANDARD CHARACTERIZATION/STABILITY AnalyticalReference Standard: Common Name: Physical Description: Lot Number: Storage Conditions: `Expiration Date: Date Received: Amount Received: Supplier: ICntoemrnmaol nStNaandmaer:d: Physical Description: Lot Number: ESxtpoirraagteiCoonnDdaittei:ons: Date Received: `Amount Received: Supplier: DMeLv-aMleovnailconAiccidAcLiadctLoancetoorne Mevalonolacetone or MVL `White crystals 47300/1 50498 543 C, desiccate, store under nitrogen `September 13, 2002 (Assigned by PSreipmteedmibcear) 13, 2000 S grams Aldrich Chemical Co. DMLV-LM-edvaylonolacetone-4,4,5,5-ds Clear liquid U-295 2Se2p4t5emCber 11, 2002 (Assigned by Primedica) September 11, 2000 0.1 gram Cambridge Isotope Laboratories Characterization and Stability: The characterizationofthe analytical standard and internal standard is the responsibilityofthe Supplier, as is the methodofsynthesis, fabrication or derivation and stability determination. 418.018:PAGE 1-10 GPRIMEDICA Primedica-Worcester Project number: PABX-BVA Page9 Final Report For Pimedica Argus Study Number: 418-018 ARCHIVAL STORAGE Archival Storage: The original Final Report and raw data will be maintained for a minimum period of five years following submission of the final report in the Primedica-Worcester Archives in Worcester, MA. After five years the Sponsor will be contacted for disposition instructions. Archival material will be indexed by Primedica-Worcester Report #PABX-BVA-01-62. 418018:PAGE L-11 SPRIVEDICA Primedica-Worcester Project umber: PABX-BVA Page 10 Final Report For Primedica Argus Study Number: 418-018 ABSTRACT: A procedure has been developed and validated for the determination of mevalonic acid in EDTA rat plasma. The procedure involves the conversion of mevalonic acid (MVA) to cmehvraolmoantiocgraapcihdy lcaocutpolneed (wMiVthL)maasnsd stpheectarnoamleytsriys (oGfCa/MlSi)quiuds/ilinqguipdoseixttirvaectchbeymicgaals ionization (PCI). A mathematical factor to convert sample concentrations of MVL to MVA is included in the Laboratory Method in Appendix A. Calibration and quality control standards are prepared in water since mevalonic acid is a naturally occurring compound in plasma and concentrations vary with diet. The internal standard, MVL-ds, `was used for quantification. The results for the validation indicate that the method is sufficiently linear, specific, reproducible and accurate to support the analysis of mevalonic acid in EDTA rat plasma. samples. 418-018:PAGE I-12 SPRIMEDICA PrFiimnaeldiRecpao-rWtorcester Project number: PABX-BVA For Primedica Argus Study Number: 41P8a-g0e1811 INTRODUCTION: The objective of this study was to develop and validate an analytical method for determining levels of mevalonic acid (MVA) in EDTA rat plasma. EXPERIMENTAL DESIGN: Overview: The assay described in this report employed the conversionof mevalonic acid (MVA) to mevalonic acid lactone (MVL) and the analysis of liquid/liquid EDTA rat plasma samples by GC/MS using positive chemical ionization. The internal standard, MVL-ds, was used for quantitation. The validated assay covered the concentration range of 10.0 ng/mL to 250 ng/mL of MVL in EDTA rat plasma using 100 uL sample volumes. Materials and Methods: Refer to Appendix A for a detailed description of materials and methods employed during the course of this validation study. In addition, quality control (QC) standards were prepared in water at the lower limit of quantiation (LLOQ QC, 10.0 mg/mL) using the same stock solutions used for the water QC standards. A set of QC standards (Low, Mid and High) was prepared in rat plasma using the same stock standards and dilution `procedure used for the water QC standards. One set of solution calibration standards was prepared to mimic final extract concentration assuming 100% recovery using the same stock standards used for the calibration standards. General Comments: Representative raw chromatographic data from validation day 1 are provided in Appendix B. Validation Design: Validation Description: `The assay was comprised of five analytical runs. Three of the five analytical runs were comprisedofbracketing seven-point water matrix calibration curves with six replicates of `water matrix quality control (QC) samples at four concentrations, six replicates of EDTA rat plasma matrix QC samples at three concentrations and six replicates of rat matrix control samples (EDTA rat plasma with internal standard). Multiple water matrix blanks and water matrix control samples (matrix with internal standard) were analyzed in each 418-018:PAGE 1-13 PREDICA Primedica-Worcester Project number: PABX-BVA Page 12 Final Report For Primedica Argus Study Number: 418-018 run. An analysis of the method recovery was included in the third analytical run, which consistedofan extracted calibration curve and a solvent calibration curve. The analysis of freeze/thaw (F/T) stability and room temperature (RT) stability, both in water and matrix, was included in the fourth analytical run. The fifth analytical run was a re-injectionofthe 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 | Validation Course - Day? Days EE `Precision, accuracye,quliinveaalreitnyc,yand water/plasma Precision, accuracy, lncarly, specificty, and waier/pascna| equivalency Precision, accuracy, linearly, recovery, dillon aceuracy,| precision. and waterfplasma equivalency. rr Parameters Evaluated: Mevalonic acid is a naturally occurring compound in plasma. The concentration levels found in blank plasma were anticipated to be above the validated lower limit of quantitation (LLOQ) of 10 ng/mL. This level of background concentration would significantly skew the results of plasma QC's at the lower concentration levels. Therefore, calibration and QC standards were prepared in water matrix to evaluate precision, accuracy and linearityof the method. The second setofQC's was prepared at three concentration levels in EDTA rat plasma to evaluate the ability of the method to extract and quanitate MVL equally from water or plasma matrices. The plasma QC's were also used to evaluate matrix stability. 418-018:PAGE I-14 PRIMEDICA| PFriniamledRiecpao-rWtorcester Project number: PABX-BVA For Primedia Argus Study Number: 41P3a-g0e1813 Water/plasma extraction equivalency was evaluated by the analysis of six replicates of each QC standard concentration levels in bothwaterand plasma. Mean peak area counts of the isotopically labeled mevalonic acid lactone intemal standard were calculated. The difference between the water and plasma mean peak areas were expressed as percent difference. The acceptable limits were 10% difference. The isotopically labeled internal standard was used to evaluate extraction equivalency because it is chemically identical to mevalonic acid lactone and is not present in blank plasma. Therefore, the intemal standard mimics the extraction behavior of mevalonic acid lactone but has no naturally occurring background concentration to interfere with the results. `Water/plasma extraction equivalency was also evaluated by the six replicatesofHigh QC standards in both water and plasma. The endogenous concentration of mevalonic acid was low compared to the high QC concentration and did not make a significant contribution to the concentration results. Mean concentrations of mevalonic acid lactone were calculated and expressed as percent bias compared 10 the nominal concentration. The present bias values from the high QC in plasma were compared with the water QC `and expressed as percent difference. The acceptable limits were 10% difference. Inter- and intra-day accuracy and precision of the method were evaluated at four concentrations by analyzing six replicatesofthe QC standards prepared in water over the courseofthree validation batches. Accuracy results are reported as % bias. Method precision for QC standards is expressed as% variance. Inter-day method precision and accuracy were also evaluated for calibration standards. Dilution accuracy and precision was measured by analyzing six replicate dilutions of a QC standard diluted 1:5 water. ~Intra-day precision and accuracy was evaluated and reported as % RSD and % bias, respectively. Recovery of the assay was evaluated by comparing the absolute peak areas in extracted standards to those obtained from solvent standards prepared to mimic final extract concentrations assuming 100% recovery. Linearity of the method was evaluated by visual inspection of the calibration curve and examination of the correlation coefficient (r) for each standard curve. Freeze-Thaw (F/T) stability in both water and plasma matrices were evaluated by analyzing six replicates of quality control samples prepared at two levels, which were subjected to three F/T cycles, prior to their extraction and analysis. 418-018:PAGE I-15 PRIMEDICA Primedica-WorcesterProject number:PABX-BVA Page14 Final Report For Primedica Argus Study Number: 418-018 Room temperature stability in both water and plasma matrices were demonstrated by analyzing six replicates of QC standards prepared at two levels which were leftstanding at room temperature for seventy-two hours prior to their extraction and analysis. Six-day room temperature stability of final extracts was evaluated by re-analyzing the day-one validation extracts consistingof duplicate calibration curves, six replicates of QC standards in both water and plasma matrices. Assay specificity with respect to endogenous compounds was evaluated by assaying six unique plasma lots from untreated rats. RESULTS AND DISCUSSION: Validation Results: Water/Plasma Extraction Equivalency: The ability of the method to extract and `water or plasma matrices was evaluated. quantitate mevalonic acid equivalently from The equivalence was evaluated by the analysis of six replicates of eachofthree QC standard levels in water and plasma during the first three validation runs. Mean peak area counts of the internal standard (isotopically labeled mevalonic acid lactone) found for the plasma matrix QC's during each run were compared to the results for water matrix QC's. The difference was expressed as % difference. Percent difference values for the first three validation runs were 0.5%, 0.5% and 4.4%, respectively. Refer to Table 13 for a complete summaryofresults. `The comparison was also evaluated by the analysis of six replicatesof high-QC standards in water and plasma during the first three validation runs. At this concentration level (200 ng/mL) the endogenous level of mevalonic acid was not a significant interference. Mean concentrations of found for the plasma matrix High-QC's during each run was compared to the results for water matrix High-QC's. The results were expressed as %difference. Percent difference values for the first three validation runs were ~2.0%, 0.5% and 0.5%, respectively. Refer to Table 14 for a complete summaryofresults. The percent difference results were all within the acceptable limits of 10%. The extraction of mevalonic acid lactone from water and plasma are equivalent. Therefore, 418-018:PAGE 1-16 PRIMEDICA PFirniamledRiecpao-rWtorcester Project umber: PABX-BVA For PrimedicaArgusStudy Number: 41P3a-g0e1515 the calibration curve and quality control sample prepared in water can be used to quantitate mevalonic acid lactone concentration levels in plasma. Accuracy: Inter-day and inira-day accuracy were acceptable for the assayof Mevalonic acid lactone: in water over the concentration range of 10.0 ng/mL to 250 ng/mL. Intra-day accuracy was evaluated by the analysis of six replicates of each of four QC standard levels in water during the first three validation runs. Mean concentrations found dreulraitnigvecatochthreuonrewteicrael cnoommpianraeld ctoonctehneotrreattiicoanls nwoamisnaelxpcroenscseendtraasti%onsbiaansd. thPeerdciefnfterbeinacse values for the lower limit of quantitation (LLOQ-QC, 10 ng/mL), low-QC (25 ng/mL), `middle-QC (70 ng/mL), and high-QC (200 ng/mL) standards ranged from 4.8% to 0.1% over the three analytical runs. RefertoTable 1 foar complete summaryofresults. Inter-day accuracy was evaluated by the analysis of six replicates of each of four QC standard levels in water during the firstthreevalidation 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. Referto Table 1 foar complete summaryofresults. The interday 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 Tabl2e for a complete summaryofresults. Precision: Inter-day and intra-day precision was acceptable for the assay of mevalonic acid lactone in water over the concentration rangeof 10.0 ng/mL to 250 ng/mL. Intra-day precision was evaluated by the analysis of six replicates of each of four QC standard levels in water during the first three validation runs. The intra-assay variance estimate of the interpolated concentrations for the replicate QC standards ranged from 1.3% to 4.5% across concentrations. Refer to Table 1 foar complete summary of results. Inter-day precision was evaluated by the analysis of six replicates of each of four QC standard levels in water during the first three validation runs. The inter-assay variance estimate of the interpolated concentrations for the replicate QC standards ranged from 0.4% to 1.2% across concentrations. Refer to Table 1 for a complete summaryofresults. 418-018PAGE 1-17 JPRIMEDICA PFirniamledRiecpao-rWtorcester Project number: PABX-BVA For Primedica Argus Study Number: 41P3a-g0e1816 `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 summaryofresults Dilution QC: `The dilution QC results were acceptable. Six replicates of the dilution QC with a predilution concentration of 200 ng/mL, were diluted with water by a factor of 5 prior to extraction and analysis. The mean concentration found was compared to the theoretical nominal concentration (40 ng/mL) and the difference relative to the theoretical nominal concentration was expressed as % bias. The result was --1.8% bias with an RSDof3.0%. See Table 8 for a complete summaryofresults. Recovery: Recovery values were calculated by comparison of mean duplicate spiked and extracted calibration standard peak areas in water to the peak areas obtained for solution standards prepared to mimic final extract concentrations assuming 100% recovery. Recovery ranged from 32.9% 0 39.6% for mevalonic acid lactone and from 33.6% to 42.0% for the intemal standard across concentrations. Refer to Tables 3 and 4 foar complete summary of results. Linearity: The assay was linear for mevalonic acid lactone within the calibration range of 10.0 ng/mL to 250 ng/mL. Fspiigkuerde c1alsibhroawtisonthseta1nd/aXrdwdeaitgahtpeoidntlseafsotr-osqnuearoefs tlhienevaarlirdeagtrieosnsriuonns.ploLtinweiatrhitythweasactaulaslo indicated by the correlation coefficients (r) obtained, which were greater than or equal to 0.998 for each of the first three validation runs. Refer to Table for a summary of regression constants. Specificity: Mevalonic acid is a nawrally occurring compound in plasma. The background concentration level in the plasma QC's was evaluated by the analysis of six replicates of control blank EDTA rat plasma matrix during the first three validation runs. The inter-assay mean concentration found was 16.6 ng/mL. Refer to Table 11 for complete results for the control blanks of plasma. 418-018:PAGE I-18 PRIMEDICA PrFiimnaeldiRecpao-rWtorcester Project number: PABX-BVA For Primedica Argus Study Number: 41P8a-g0e1817 The variation of background concentration levels in EDTA rat plasma was evaluated by the analysisofsix unique lotsofplasma during validation Day 2. The result was a mean concentration of 15.8 ng/mL. See Tables 12 for complete results ofthe six unique lotsofplasma. Stability: The acceptable criteria for stability evaluations were a mean bias of <15% from the theoretical values except for the lowest calibration, Low-QC and LLOQ QC standards, where the criteria were 20%. Due to the presence of background concentrating of `mevalonic acid lactone the theoretical values used for the Plasma QC standards were the mean concentrations obtain during the same analytical run from the analysis of six control Plasma QC standards. Room temperature matrix stability was evaluated by analyzing six replicates of quality control samples prepared at two levels, in both water and EDTA rat plasma matrices, which were exposed to room temperature for 72-hours prior to their extraction and analysis. The values for % bias were within the acceptance criteria for both concentrations for the 72-hour room temperature stability assessment. Refer to Table 6 for a complete summaryofthe results. Freeze/thaw matrix stability was evaluated by analyzing six replicates of QC standards prepared at two levels, in both water and EDTA rat plasma matrices that were subjected to four freeze/thaw cycles. The values for % bias were within the acceptance criteria. Refer to Table 7 for a complete summaryofthe results. Six-day room temperature stability of final extracts was evaluated by re-analyzing the duplicate calibration curves and the six replicates ofQC standards, in both water and EDTA rat plasma matrices, from the first day of the validation. The solvent in the extracts evaporated from the vials during the storage period. More solvent was added to each vial to reconstitute the extracts. Results from the re-analyses demonstrated that the extracts were stable for a minimum period of six days when stored at room temperature. See Tables 9 and 10 for the final extract stability results. Studiesoflong-term storage stabilityof mevalonic acid in matrix at -20C are ongoing. 418-018PAGE 119 OPRIVEDICA PFirniamledRiecpao-rWtorcester Project number: PABX-BVA For Primedica Argus Study Number: 41P8a-g0e1818 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. Primedica-Worcester Projet number: PABX-BVA 413-018:PAGE 120 Page 19 FIGURE imeiWorcs Poet he: PABXBVA suas Day ValinCaltrainCurse 418-018:PAGE 1-21 reg2e0 , " 1. LgSe F o.# w0l4] a. VeVseresmfsaPioss A | _--_ || _-- + swore ||1 -- Sogomore Yeint: 0.011375 TR----_ T-- E d CommntRitPitss Primedica:Worcester Project number: PABX-BVA 413-018:PAGE 122 Page21 TABLES Primedica-Worcester Project number: PABX-BVA 418018:PAGE 123 Page 22 TABLE | SUMMARY OF INTERPOLATED MEVALONIC ACID LACTONE QC STANDARD `CONCENTRATIONS WITH INTER AND INTRA-DAY SUMMARY STATISTICS Day 100"Theoretica2l No0minal 00 Concentrations (ng/mL)00 RReepp2 589613 225306 01 687 0 20 Rep3 551 250 8 196 RReepps 903785 223490 66734 11995 Reps 992 239 3 199 Meaan 9362 252 67 1598 Bias ame am am 0% Day2 100"Theoretical 2No5m0inal 00 Concentrations (ng/mL)m0 Repl 087 204 0 RReepp3 190239 242418 6a814 119955 Rep4 007 23 82 193 Reps 910 2438 0 194 Rep 6. 102 245 67.3 195 Mean 964 24 683 196 "Biaas S6e aa5n 24 % 205 % Primedica-Worceste Project umber; PABX-BVA 418-018:PAGE 124 Page23 TABLE 1 (Concluded) S`UCMONMCAERNYTROAFTIIONNTSERWPIOTLHAITNETDEMR-EVAANLDOINNITCRAL-ADCATYOSNEUMACMIADRQYCSTSATTAINSDTIACRSD Day3 `Th0eo0reticalNo3mi0nalConcemntorations (ngmiwmL) RReeppl2 oorm 2229 61 118 Rep3 os 24 06 20 RReeppsd o9s%e a2u1s6 000 11%8 Reps NA Bs 60 1s Mean ol ue 1 107 %Binas 225% e5n 06% L6s IInncnraassssayy vvaarriiaannccee ccssiimmaattee 4152%% 2057%% 1132%% 0148%% Intrday Mean "Bias 1os7e 204) @108 11:8 a6 2a aw sw NA = Not Availabe. This sample was accidnally spilled during preparation Primedica-Worcester Project number: PABX-BVA 413-018:PAGE 1-25 Page 24 TABLE2 SUMMARY OF BACK-CALCULATED MEVALONIC ACID LACTONE CALIBRATION STANDARD CONCENTRATIONS `Theoretical Nominal Concentrations (ng/mL) 100 200 30 50 00 10 20 Dayl 9% 200 307 06 895 151 254 01 200 300 497 81 144 254 Day2 98 195 296 S04 908 152 250 03 193 36 @2 97 us 250 Dy3 103 197 05 S17 917 17 21 954 209 06 8 848 us 251 Men 100 199 305 499 $94 148 253 SdDev. 0296 056 0681 134 266 30a 418 n 6 6 6 6 6 5 6 URSD 30% 28% 22% 27% 30% 20% 16% %Bias 00% 0S% 17% 02% 06% 16% 13% Primedica-Worcester Project number: PABX-BVA 413-018:PAGE 1-26 Page 25 TABLE 3 MEVALONIC ACID LACTONE EXTRACTION DATA & RECOVERY CALCULATIONS Extracted Standards Solution Standards MeanResponseAreas ResponseAreas sl ans 1066 s2 785 1981 s3 10575 2963 Sus 1396 975 sus 3144 3678 sub 46935 19279 su? 3433 25380 Mean Std. Dev. %RSD % Recovery 392% 39.6% 357% 311% 362% 20% 32% 364% 00327 24% Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-27 Page 26 TABLE4 INTERNAL STANDARD EXTRACTION DATA & RECOVERY CALCULATIONS Extracted Standards Solution Standards MeanResponseArea ResponseArea ssuul2 56817462 15019 14959 s3 5365 15158 ssuuads 65060547 14283 14739 sus sis 15226 sa? ss 15286 Mean Std. Dev. %RSD % Recovery 09% 39.3% 35.4% 20% 34% 336% 345% 37.7% 00327 87% Primedica-Worcester Project number: PABX-BVA 413-018:PAGE 1-28 Page 27 TABLES . SUMMARY OF LEAST-SQUARES LINEADRARTEEGSRESSION CONSTANTS AND ANALYSIS Slope Dayl 00065514 Day2 0.006308 Day3 00062253 Dayd 00062800 Days 0.006156 Imexept 00113750 00082756 00067910 00060925 00112850 ComelationCocfliient() ~~AnalysisDate: 099975 1017700 099939 1018/00 0.99834 1019/00 099891 102000 099981 1023/00 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 129 Page 28 TABLE 6 SUMMARY OF QC STSTAANBDIALRITDY7I2N-MHAOUTRRIRXOOM TEMPERATURE RReeppl2 RReepp3 Reps Reps Mean Std.nDev. %URBSiaDs WATER QC's `Theoretical Nominal Concentrations (ng/mL) 250 Ei 22318 119819 223938 119933 230 194 29 194 233 192 04632 1697 6198%% 4100%% EDTA RAT PLASMA QC's Theoreti3ca4l20Nominal Concentrat1i0o2ns0(ng/mL) Repl 351 199 RReepp23 334420 210907 Rep 32 Reps 36 119954 Rep6 3s 188 Msde.aDnev. 0363794 413926 n 6 6 %RSBiDas 0290%% 2212%% += Mean concentrationofsix contol Plasma QC's analyzed i the same analytical batch. Primedica-Worcester Project number: PABX-BVA. 418-018:PAGE 130 Page 29 TABLE? SUMMARY OF QCSTSATBAINLDIATYRDINFMRAETERZIEX/THAW (3 CYCLES) WATER QC's Theoretical Nominal Concentrations (ng/mL) 250 20 Rep 23 194 Rep2 256 194 Rep3 22 194 Rept 28 194 Reps 20 196 Rep6 23 192 Mean 25 194 Sud. Dev 04m 126 n 6 6 RSD 20% 06% % Bias 6.0% 2.0% EDTA RAT PLASMA QC's Theoretical Nominal Concentrations (ng/mL) 420 Loe Rep! ns 194 Rep2 344 200 Rep 29 197 Reps 336 193 Reps 23 190 Rep6 24 184 Mean 32 193 Su. Dev 0804 559 n 6 6 %RSD 24% 29% % Bias 29% 05% + = Mean concentrationofsix control Plasma QC's analyzed inthesame analytical batch. Primedica-Worcester Project number: PABX-BVA. 418-018:PAGE 1:31 Page 30 TABLES SUMMARY OF INTERPOLATED MEVALONIC ACID LACTONE DILUTION QC CONCENTRATIONS FOR DILUTION VALIDATION Theoretical Nominal2C0o0ncentration (ng/mL) Repl 04 Rep2 305 Rep3 07 Repd. 381 RReepp6s. 389 379 Mean 393 Std. Dev 116 n 6 RSD 30% % Bias 18% + = Dilution QC undiluted concentration was 200 ng/mL (1:5dilution performed prior to extraction and analysis) Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-32 Page 31 TABLES 6-DAY ROOM TEMPERATURE FINAL EXTRACT STABILITY BACK-CALCULATED CALIBRATION STANDARD CONCENTRATIONS `Theoretical Nominal Conceatrations (ng/mL) 100 200 00 500 200 150 20 987 198 02 502 899 150 254 102 197 305 04 906 146 253 Men 100 198 304 08 903 148 254 a 2 2 2 2 2 2 2 %Bias 04% 3% 12% 04% 03% 13% 14% Primedica-Worcester Project number: PABX-BVA 418.0I8PAGE 133 Page 2 TABLE 10 6-DAY ROOM TEMPERAT`SUTRAENFDIANRADLCEOXNTCREANCTTRASTTAIBOINLSITY QUALITY CONTROL RReeppl2 Rep3 RReeppss Rep6 Mean SDev. "RS D wBiss WATER QC'S Theoretical Nominal Concentrations (ng/mL) 100 250 200 200 501879 22531 1687s 119988 105 35 a7 197 00715 u4e1 ae3a 210908 101 243 699 199 00 247 ess 198 0S 042 086 103 s6s 206% Asm o05sw 00% A2% 20% -L0% EDTA RAT PLASMA QC'S 81 `Theoretical ne Nominal Concentrations (1n0g4/%mL) Repl 350 ns 195 Rep2 344 756 19 Rep3 28 762 192 Reps 22 ns 191 Reps 316 707 186 Reps. EY 749 185 Mean 33 57 190 Su. Dev. 131 226 378 a 6 5 6 %RSD 39% 1% 20% 4Bias 06% 03% 21% + Mean concentraioncsv onthe rst dayofvalidation Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1:34 Page33 TABLE 11 SUMMARY OF CONTROL BLANK EDTA RAT PLASMA CONCENTRATIONS Repl Rep2 RReepps3 Reps Rep6 Mean Sd.Dev. n RSD. InterAssay Mean Std. Dev. 4RSD Concentrations (ng/mL) Davl Dav2 Dav3 164 165 170 162 159 168 166 163 168 m4 174 173 163 161 171 158 162 170 165 164 170 05% 0320 0.190 5 32% 326 % 116 % 166 18 0506 30% Primedica-Worcester Project number: PABX-BVA 418-018PAGE 135 Page 34 TABLE 12 SUMMARY OF UNIQUE LOTS OF EDTA RAT PLASMA CONCENTRATIONS `Concentrations(ng/mL) LLoot2t 116120 Lots 163 LLootts s 116716 Lot6 170 StMd.eDaenv. 214518 "4RnSD 1563% Primedica-Worcester Project number: PABX-BVA 418-018PAGE 136 Page 35 TABLE 13 SUMMARY OF INTERNAL STANDARD PEAK AREAS Water QC's Davi Dav2 Davd LowQC RReeppl2 4038906 a4s30l3 SE1n1 ReRepp3 5031608 sssis "s0s6s RReeppss. pair2 6s463270 2380436 MidQC RReeppl2 4a3920 sawso 0522380 RReepp3 aam2 Ss7m57 4s7es9 RReeppss 01244 6s6a3s2t s5i95620 HighQC RReeppl2 asiosn s0a1n5s 511947 RReepp3d 2350668 53092590 6s1o8798 RReepps6. 5si3s1t S200207 7e638345 Memaea a821 sn sem Primedica- Worcester Project number: PABX-BVA 418018:PAGE 1:37 Page 36 "TABLE 13 (Concluded) SUMMARY OF INTERNAL STANDARD PEAK AREAS LoweC MdQC HighQC Plasma QC's Davl Dav2 Day3 Repl a0 as03 6100 ReRepp23 a3850 5m3663 s5i0s6t7 RReeppds S3038540 S18540 55910599 Reps sis ES 6755 RReeppl2 13598808 0S19517 s6a0n4z0 RReeppt3 w2r5 52063 166 RReepps6 s08i8 SSo0u1t 6s4n9d Repl 6540 nas so8s Rep2. 5208 5474 5811 RReepp3 570002 6a08t8 65068145 RReepps6 s3e5n08 7san3t 5547697 Menaws 447 57 sor `Comparison of PlasmaOCvsWaterOC MeanAvess "Difference 0.5% osw 9 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-38 Page37 TABLE 14 SUMMARY OF PLASMIANTQRCAS-DTAAYNDSAURMDMCAORNYCSETNATTRIASTTIICOSNS WITH INTER- AND Davi Repl Rep2 Rep3 Repd. Reps Rep6. Mean n % Bias % Differ(eTnacbelves1W)ater QC Dav2 RReeppl2 Rep3 Repd. Reps. Rep6 Mean n %Bias % Differ(eTnacbelves1W)ater QC `Theoretical Nominal Concentrations (ng/mL) 250 00 20 3s 757 198 na 73 191 24 753 192 50 ns 199 322 32 27313 119930 a1 79 194 6 6 6 24% se% 30% 20% `Theoretical Nominal Concentrations (ng/mL) 20 0 00 343 764 198 n2 759 194 30 769 202 34 753 194 348 783 194 351 74 199 340 769 197 6 6 6 360% 95% 15% 05% 418.018PAGE 139 Primedica-Worcester Project number: PABX-BVA Page 38 `TABLE 14 (Concluded) SUMMARY OF PLASMA QC STANDARD CONCENTRATIONS WITH INTER- AND INTRA-DAY SUMMARY STATISTICS Davi Repl RReepp32 Reps RReepns Meaa n % Differe%ncBeivsssWater QC (Table 1) 20 `Theoretical Nominal 0Concentrations 2(n0g/mL) 349 755 198 335415 L7a68 119097 2 m9 195 338477 746.70 210925 169 36 198 396% 0% 1050%% Inotterra.-assssaayy vvaarrioanncceeeessitimmaattee 22.34%% 2108%% 01%% Totr-Asay Mean Biss 31480 715s7 119s6 360% [A 20% Primedica:WorcesterProject number: PABX-BVA 413-018PAGE 140 Page 39 APPENDIX A LABORATORY METHOD PrimediaWorsePct ber: PAB BVA JEDI corronaTIoN eA Spier 418-018:PA1G-4E1 voged0 vente vais r r--RRRL er 1 or 1 somes JN72dP<)D. EI) ves (osin Sly Authorized By: ErAN ci ue _ultfoo owe silos vue _12 [5 Joe Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-42 Page dl FE ARATE SEVALONIEACID TN RAT FLRSA BY GO HNevsie n NombeSr:A8L FPopeietD1e2 Nerve Too 1 Purp ConencpeunrtpaoisoenosfofhismLeoirnysiMethodp(aLnM)sis0adesmcrbipybeGlpCrMeoSDcs.eso scoraly decmin he 1 Scope ToPTaeactsposnreo(EcDMuTYrALe.)s TpDrhaoreviivdnledihieocextdtiasccLtoMolnrimcenpvparllaoinncigcsesicio1g00ihmIeVgaAumaci1foi0nc2vt5ieornnotmfe.mdeovmfamlaoenlilcoonsd.cinirt Iaionrn inpttpmlEoDTEDATAs:iTghtehrcemointvcertbsdceocnrvieedne ohehcu oonfpt srcoimon.ofevan 3 DefintionsAbbrevitions cwics: 1s MGIaanssiCaSheSeurinndoearDmd eosc jivae: MMeevvalioonnicc Aaccidd) coDLnMeleoni Acid actoneor MvLa; DMIoMevmoiotnoomcined 45540 oGc QGuraavyiCono)moCorson 4 Meri a1 remiss pDDLLu-MMneeavavonsacciodncei.n4.e,i54,gCnaCmbhriidgce eCot. parLoaxibmoarla97,6 g38d% roarde, pEMyioiasncntenhe,i7.dBeak.er, BHaPkeLrC, WgPraLdCegorraccqeiovrlsenutvalen nFaorpnainoalcsda)proByok!ea,ccoag,egEMmScoernce,pHPlLC gree or spielen CShoidooimonumf,o1,7ABVakIer.. HrPeLgCesnnrdacoerosrqcuiivlaetn DcehiounirzeNdowrobeeo,stMiAlvlgoer.e Mil. lorawiaeh,porcyiofaqlue Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-43 Page 42 RE ARATYSR MEVALONIC ACTTNRAHT PEASHA BY GONSD" TWeeviaiovneNurmbeSr: A0T EPafc erotDr12e November 700 HIseorbsuntean,e, M-aT. tBheesro,nHGPsLCPrroaddesorrseisvearlnratde ce equivalent Pooled i lok pas EDTA 42 Consume DPiispnosaanblen1d3pmsl.conc)bovompalypeopyene et bes DEGipsippeaonscdirobiltmpestpeooppelaysettsyelnsndmsptlasesrtierhpwit pDotesbqesnievcsalec snt ot fcqvs alen Polypropylenevise wis ing sid aps o valet 43 Lapmen VLaObRndMulsttii-eTsuPbeptVeDirsspeensoerrocrqicviavllenen. iZeackkmanTGbSov6sKpRLcVeErvafpgaoreraissoueapsvlnt AJANWDGERC-1c8o2mAes,aDBy-s21]0b,t3n0mc,eo0.r25cmint1e, 0.25 Fil ickneosrequivlen HHewelettt PPaacckkaarrdd 668930gmsccrhroomqoiapahncrcqivalen Heevwilcet PPaacckkaarrdd C$9h7e3mrSuisniosenle(cGti1v7e0d1eBtAe,ctRoerovirsieonqu0a1l00) or eile Micros Exce7l version SK-2orcite sNotye: pEeqrufiopmmaenntc.eesSpenmso.neco.nsumablceasnbesubst poviddht souvent 5 Procedure To1ht5i1sp0ml0eamtnphaomEd.DhT1a25A0heby GlmEdlMs.SteoDfdvmeiovrnaglhoe1ns i0c0acisdl aosmfcpmlieeovnvas olpohnmliecsesshThee0vEmDaeTvlAa.leonTichcoecondmcveceirrnnie)rooanfninge iSreieor.nboetelpobl.oN.oeeMloaotvftimhenl6obndrsa43sniadcrunrolvety3pdolcqcusuarliiEtnyDgTccoAonmvoplocssetianrdpasrdlsoshaec3a1lp0rcelpaairoednsin rConmgecetmoenthwoidllailyswiinhwdiet.r sCopnocseren1f7oinndk.oTchids tcskgorfobuonnd coancepntlrastioEn DevTeAl Seid nef wilhthquanta. specily he ows calito evel This method bss + emovnasdiash1etvtieeweatveramnixccdSrcoonne Scuarvecs uantdeqeusaldlistyoCarowltpeitsi.nThceovuvlalannt ra pags EDTA. `Primedica-WorcesterProject number: PABX-BVA 418-018:P1A-G4E4 Page43 e[ mwr -- erat e tptmy gSs. I w S C= o eaE rer: 12 rS m--m--oe at--sp ----ehteeea edost i r Si E Se Bo TOhSTie,Seenaaurt or e i a --P -- aCoe AmT ep Le 15% DpottemesotmSpapdmtcsemnitmee53 es tthe 5 pPpeetceipveTsS neirE i E E P y e T es rete, A i 1 Wms `Primedica-Worcester Project number: PABX-BVA 418.0I8PAGE 145 Pagedd TERSAC SIS EVATONE RO IWATPEASY BY Ges NCeo mNameSr:A0T SpTase St 11 Re oNontelhtyMYL hiooeopS.lAtgofesmhoooPufOMRnYTLEisovoedfand51.swresheed tet aan ie ermi eg Wcheitmheout passks.Brn 1v0o0lmarg oif lVcLhstnacrk tndn.to 3Th1e0s0amn.i WL commonofthson 100 pm 532 SecondaryStock 8) Tt hinnsea h753a m1v L5w0iolfho he ro marrytStmn ocdk (i4)n he2s9o0mamli.Me VEemrsk.tof 533 pationofCatenion Sandan Spin (C59)Sons Tmhiebfcolnoowrisngs0anhdrprSpueaieonsocmheounec PeheAPPINwil e20Cnmuoeatodnebrawooidnyts.peO.hreoaonkg AbD syenddrdihasbrecnowimbHed5 534 PAhTrbveoaC5r5ansalaoroodrusght2roe50mR.orcswli5kmroicl iks.sshFoolvneoToAlfeuTerok.3 ined nd htt epson rer kumi me oy PRB EL E -- FontVsSoomma] E FimnsoSoo-- mon| SCGoncTemiSraotonn iy mi) nmi [C[C CESSSTS SSeceonedamrySSwoakn[|O 168 1 sisa o [ae s s e Tao hw ow ]| CE0Semmian So |_ -- ase-- [| s--eow--]| ICC57] Fam soa|e|eCS o asen w1m]| 53.5 OT nce arT emar 2Y a0lmimhcwehoaCma)iirofoPSptihnSorlos (uggrcseedp18 56 PuepaatonofCalfenion andr (STD)in Wate Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-46 Page ds THE ANALYNEVA ONIC ACID TR RAT FLASSA BY GORD RTevbioHn NumbeFr:A0L pEasee ootDR32E Nove Ptreeprareincdaivbidruailolyswainhda0s0ds25o0tmhLdoyf ohfen spectil vbeyCcoomlby iinntionng p2k.i5nmglSoofoMtiIonL(OCSS) Toll by mining. The fl concenaon in wr se he able blow. [SCuornadtaornd]| VWoaltuemre | SVpoiliunmgeSAodudteodEn T 1 SComicemSraoioonn [Sr| zomsy wnots eT[omniTo] [SLTFs[5s ToCmE s ess0 s2wBT O mo]| [[sCmses zzms Toomms |cGeas [ssweeT Tw mo o] ] C[CsomreTaams 1 ooom Tovser| |i sm 3w0w]] 55 Preparation of QuyContl (0) tockSoltions "mTihclaonwoinnsgoa7n6adchathpereppaeraetidonnheem)eC3eSmsYggTcIiAeTdSssperoRsche.PAeppardapwiaelbe o1c0u0%mpeanried.Si tahnedaprdewtisshtavbd. aSrnincdrdrrefcerencoermpausr. Aalrl asnsdna s0olubtions hlelcueinsmiron1d2t0oftssaeressooornd.tere. Ongoing ASD30sveCambang 551 PriOCmSoackr(Ay) NorotielchlaatyMcVosLkcisshiycaeonscaof npdic.Us nAsrefnmtsob4uo0vfnMhVtr3,OiscoEu aRofndit.weSitgheeed Woeimghecnt sHpakp.roxTiimntglyo1v0a0rm owfMiVh Lctsln aacnetesnd ti.o 3Th1e00nin SVT concentofthis lon 1000 im. 552 Secondny OC Sock(8) Bs ing 10m $vo0l0ummeoe ifhoahPrsciemOCaanSdrtomycixk.(ATAh)eionress2]50 MmVlL cmoanticcenfsof solution 200 pL. Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 147 Page 46 T ReviG e NumberA:T0O 56. PreparationofQuality Cone SiinSoltis(QCSS) heoeiccfoaluiloowmsinsaeaendcnahrdiphernpaseatrbmicoo.nspAcolhlna)naC3nodsnogcgouesrnd sipbeeechp.srhAepeprorpdr2i0wsielCbweish Proeceopdni.GrCiSss.iOonntos SamyHs 25.0 mLvlc SHARE he ks haw pion es fhse bleteow. A Loiion lsoetihoonuscghitooohstwihocns3rce.rioFin Yhumneasrofethsseongdaeyrcontol erro propeldocumented nh oypo ors SoSiowoikon|| SSoweiaen| t|e SSowiEon Vwoyte|| a"Cmonie || rtoVnoytan [(|OoOCCCSSSSTMoEawDN|||FSSaeemcoamrndyin0o0c6cSSSiocmckkaBhaBm||]--ssoi3se| aom0ew |sms0s | ao ms__|| --zs0 gue] 57 PrionofulyConta (0) Sudo n Wot CpolmibnigneSo1l0a0iomnLo(0fCMSSl) )aswhatoewr,ndioveidaulale. woiwt.10ml. of chQuyConsol C CGonareyr o SCtooCmckeSnmerlaatmtoannm||SSmopcikt SoVlouttioomnm e|| p VoFliunmale.||QCoC] ncSeamvraatrodn [SCtainoawarosc1D oesstow[aosan s iw [| iwCiny|| magin o | igh OC | GGCeSSsG To0w300 | T10o0 0 Tm] 50.1 COncoe prempare. iegtorateOiChsrannds sdugcgoespte3d5p4porronornnelyS1F2mC. wl etofmmaes Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-48 Page 47 FE ANALYSISMEVALONIC ACIDTNRAT PLASMBAY GOD Ea Reviion umber: S58 Sample Exncion S81 cTorannrsflerba10n0.k rmofdcpalcihcasr)a,nOdvra(lctCsoonfv1o4,Smtedtahrotdsb(lupklic(ianepslahte), . PCoonlcpmriospiyonieenevtean.ed ldys)amp,les to india 15 ml.cons] bom 82 AdA50.0, ofMQ we1e0carch method irk S83 mAadhdo5d01b0 aio.fth ere) Sander Skin Sion 0 cach uh exept oh. 584 Voren thetubes fors minimof seconds 585 cAo4nt0.0mLof formicacdsnc ubesndvar for minimofa 5586 BAlelcouwnuvbeerson otftMoVnAhe1b0eMnVchL.forspproximtly 30mit0elosw ie for 5587 CAodnIt0e5.00mlof MQwater 0cachtobe ndvor fr a iim of5 S88 ALo0n0.0Noml:. ofpcihpleotrdoifsopremns0eermxcyhbaebev3indd1v0o56ixfio aemsipie. of 589 CCEpueprnretowtneheeiib2m5ea0asn0ylGsspyyip.mrpoDlxieimswaihlnietclhyaySdeomressisnnohtuehlsadvfeaopdrrmessnitemaaciychelasym.p5l0e0ruhm. 51810 Tprnsrerethp0vst(squbeeosesa)nbdyesecnarodimeewcinhdivaidlualay15ems.lconical boom, S511 SFuiasthelaoquceossleepyw1itSh siodeiumhelfate.on(sNotyee:.spAosMiEmSatSlIy 0$.1030rasmys eae 0 4 hesodium lc becoheuesxaect weight nr cal) S832 {Ahdeds10uemsLnof smetahynlevneocrhfoorrdsip.ronpannolfsteocnont.o cNohcbe3 icoemming apener may bevid103d is rage S813 aCpeprnotxiimfatoteheleyh2e8s00s0p)p.eoximacly mimeas appoximaely 3500 pm Prints ors er ABV ss 418-018:PAGE 1-49 rr we sar T emamo nm m mr oko ee rn mememen oes reaant Co Eg IEEmoe ewe re n BrSee, w p rE--Er. mo om a EET Bo eTeE sane train oi E EorrTl e=Ee EE Sow 2 Pinder ors Pr be,PAB8A nas 418-018:PAGE I-50 i a -- oH=n- ee gE5 BEps EBEr"e na f hE=S es en, i, w iF.T2 rn:a w sepwmsewwos = o Hr a E A n e me ms 10 sEa tiprnt a--on-- , I, rD EeTn T TT een on oA rS A, E indenNorse PABXBVA ----- 418-018:PAGE I-51 aa. ern w ro EE Em E e 11 e SS E =resrime rT ee 0 E vC oH en E ttn S y mt sm x ms xm + wn reerm-- scre-- AA gg sats A SaE pmpry--f--e-- t is EmiLs mTmRs iscomint PrintsVoss rc ber PABBA Paes 418-018:PAGE I-52 kppasi co mPAom ma -- Et site maetmse comer sis ---- S B BAT SRS wr oe weI n tete etostn I I Ee --, FR mn Pimedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-53 Page 52 APPENDIX B REPRESENTATIVE CHROMATOGRAMS FROM VALIDATION DAY 1 PrimiWorestrPct umber: PABXBVA 418-018:PA1-G5E4 Pages Pak raRepent fm ts -- ngs EEETT S crass wends SEm Y T = =~ Primedica-Worcester Project number: PABX-BVA 418-018PAGE LSS Page sa Quantitation Report (7 Reviewed) RDSaietkaoolneFile :"1D;3i7x\'W0sSchDtaDnAk3T0W0Aa0\cBeAr1B\3P1A5B-BVA\1-062-1\06Z1001.DOTpeeraVcioarl:: wh1 ae @ == a16 ntatReEgrTaRcEiAoTn DPaHsRa\C: WreeiEot HO\PARDKBVSALM (RTE Integrator) fo wes --_-- 1 i1= ifii IA i|| homage A et nd - e=o| nsmoildn45 a0w.0C0067 = 0 GR 0 SRL= 16 ATio Tn To To 7hvi1m 7H 1mng|}| | ~ A | onr t 135--orrOr T inane: WotT Fours -- e | A | |Vermette/iSomerN N d|| RR ak re a ak ve ike He5 TRTTER) 06210010 BABCOVAM Wed Oct 18 12:40:03 2000 rage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-56 Page ss Peak Area Report Now Ais: ose ame:e20120. a raresttame 57 comoss Commis Ime) tem Sep Primedica-Worcester Projet number: PABX-BVA 413.018 PAGE 157 Page s6 Quintitacion Report (QF Reviewed) @ iSfBaotarEaPilRe LDoI\rSSeD_EOAtTeA\iGrAD\iPARK.BVA\L-062-1\0621002.S0fpr_oaraViioarlt: 2an 4a5nxtacRagTraEtion Y Paxams: sti otpHsoos:en ns ova.n (78 Tnceseator) pm . ) | al ii | ) | | I | {nifoinns b aCe r Sp e r --rrr Tre tGoe rrRe s atReree. Es rtnttsikSrTomTreTroe= terTeeeeerr rreeeee--r]! | i | CE TR oO |sat nsr : 3on2s00 T TEhu1,ns1TTe6.0h 0 eR TR ikR ih 2 i| Jfi\ i Rw a Ta 1 T a Th ed GGaiooa.n PAEEAM Wed Oct 18 12:40:22 2000 nage 2 Primedica-Worcester Projet number: PABX-BVA 418.018PAGE 158 Pages? PoAreasRepkort nsSemeE awe osesEeitro 430 --r-- at Pa -- SAECATAPADPABXBUALIELT, ens on 5 Gompect Conmpilue lnsiol fm team tata Primedics-Worcester Projct umber: PABX-BVA 418018PAGE 159 Page ss | | | uancication sapere (@7 Reviewed) DfIaotarnaPie + :R\ESTDORNrTAh\PeABi\PnARE-BVAVI-063-1\0G21003.0S_fporasVtioaarl]: ]a3 x @ ix n 45 ErtA ageation Becamet:iare\stWo:eheoaso_seva. (R73 Tacegrator) p! y ee 1 |- i | | ep ofE RT E SB TT e|T E ETeTTER i Fons ast Ia | Resp: 567 ee ---- . | @E m iT m i iakA ikheinieh: Ta0TTeRE TE | riteonn:: oanTETLIAT 1 || IiA |i Co R a a Te TJ e TE eT Te Gace PAmEAN Wed oct 18 12:40i43 2000 rage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PA1-G6E0 Page 59 Peak AreaReport o Samana sae ou Acqa2050 son sepraeatere oaFmomee A2s 1E003,B0B0AATAPADPADCO2U1A sumreudmenaammo:r 57 omp3nie Cwoimlpeile ImWplomdp taamD ote prostTe 1016200 1240 rage Primedica Worcester Projet number: PABX.BVA 418-018:PAGE 1-61 Pages titoeton Bugore (1 Saviownt @iDLMataoePhe Yn0ri\mDDmAeTH\BAVABL-0\52ERSR S\OTGEIBAr.Deeos:viiall:14be 1o 6 saceprg acion puS rus: seatn.p EN -- = ee ee,i ey | it Cm) rf Tes 610620 6%64 6406m 670 6% E |w T TRi EE ]| | fon: 13(1 7.1861 Tm ri 1x Tw 7T% w Th Tm Tm 1 Iommi ed NL oT PLATT) a WE yy oer 8 -- ihiis ETT Th Te 1m J I Lesine 8we dh enikTETh 151 1h 4TRTE Th i 1m e Primedica-Worcester Project number: PABX-BVA 413-018:PAGE 1-62 Page 61 PoakArea Report NoStumtiasn:Pstie: oeAS cquam : ot72000 1508 esrauFemonneeASBntEsBtUDAATAPASEABOALOE ammE med Complies fn lo fm fem gis Prete rae 101623801201 FP Primedics-Worcester Projet number: PABX-BVA 418.018:PAGE 163 Page G2 Quantitation Report (QF Reviewed) @ DrE iataa FniletN D:\SRD_E DaATA\PASNPARK-BUA\I-062-1\0621005.DSmoeenrsVtioarl!: t& a FEy EA E--S-- shops sas_sva. (R13 Tncegracor) paren Ey El| if | |- i | | ow J4 | i| w ar Paa P ETTR ER a sofoonm!: 1H3i1s (7.243) || | f\ i ee LN Ea aC,er cE _ | [ ny eon: ra is --A || i i | 1A Lek 6% sk sh Sw em aw ew 2 1% 7h Tk 7m 1% Te mim 2k Geai005.D BABEVAN Wed oct 16 12:41:20 2000 sage 2 se. ar rth 418-018:PAGE 1-64 0 crateSA gen EERRrEEns mans an ReSr ee i. Primedica Worcester Project number: PABXCBVA 418-018:PAGE 1-65 Page 64 ST Jun Bie + JYpT Or ST aL ot EEL re i IR TEr EE evenansvnx we ovegeacoet Re Vo| fil || wyTy-0U0CEE-- EEERREEEETNEELRSIIiSEEod||y fIi i| N| a JNhf || @noaEk ek -ek ok 6T % 6mT aw Tk Thm7wams 1% 10%.070k 7@ 7h I767-- | LooTe aks Tan a ea m wee The 1% re 16 7h Te IB. 288 1% Primedica-Worcester Project number: PABX-BVA 418-018:PA1G-6E6 Page 65 PoakArea Report NotaSamoaenness:FsBtCeEuvn eoe: esSeti om158 nmeT Meaen CAuBSeBAhThPADOOVAALBK samen7 Rein Primedica-Worcester Projct number; PABX-BVA 418-018:PAGE 1-67 Page 66 Guaseication Report (Gr Reviewed) @iDBEa:rtEaoNnFile |H DB: SueDeONTaA\PRG\PABK-BVAVI-062-\0621007.0SfroaeVitalo: r7o #f8 o1oseorasson BasramEs: costarS. R r= TH TT |= i | | wl I ! L I we TEE EEe E RETAeAh S ga EEE | I seep: 1054 ] | i | I JN | Cree:E ae sinpS r+o i AmGu|ntT : 0.00 cir | fi | | J\ | R ER T Te ve cErovno BARXBAN Wed oot 30 13:41:59 2000 rage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-68 Page 67 Peak Area Report [SStampale:t5o7r nhece1017250 1008 ron SEAT ErU appi5n1 nS noose PrimediaWorestrPretnumber: PABX.BVA 418-018:PAGE 1-69 saoes CO, e J B=otD EE n tapell 5 I LEE EE RR even. evs eeepc i He | IA | la e T EE EEE E RE AETE E RE i iIt i SR aJ. | Lal or trae , ! f\ | ! JA | Wek sk oo sm em en ow rw io 1m 1% 7 1m rw ik vk re | Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-70 Page 69 Poak Area Report o Sengeane: com oesc or 1077 Oui Fae Name:ce2c080 rune am: 7 PrimediaWorse rjc amber: PABXCBVA 418-018:PAGE I-71 Lo meen pBararide DIVmEaRDmATe PABc\uEnRcRiKtBatAiLon06r-e1p\eGrKEIOO%0.nDT_rrevrivieiiwahs:!t 3Bae a Ew PEER ees: 2 1 rgmErcEionS pass:R matesmemes san 8 tnceacon Jan Sr = it Lo I nw EE Toes 610 62 6% 60 6% &e 6% em {Mont 131 (0.0000 Ti rio 1m 1% 1 Et eT eT i AN rT fre es EMT EE RT SagI e EN | | Te | | = i I i lee 16% ox aw em am sh em sm rm rw 7m 7 re 7m te 1m iw rm Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 172 Page 71 PeakArea Report ne sama:BEocwo oecSueas:nr 0.47 nr C edia dPamo SAWBR,OCAATASADEADLOVAL tm emo 0 Gmmedr commits Ime] Bn teg feime at tecc ta pt tertont. Foun Tn 0012.2 ve 1 Primedica:Worcester Project number: PABX-BVA 418-018:P1A-G7E3 Page 2 Guancitation Bepore (Gr Reviewed) PY BDatoarFile | DB : USDR ONTA\PASa\PABK-BYA\A-062-1\0E21010.DS_gpeortviinarl:: i1a0n 45 aceEgraRtiTon E Furuse: REIMTE.rEha oospassava (kre Taceseaton) oor ---- - | N | F|o iI || | J4 | fimersm6104 SH 4B SR ToE ER ETRRiw SthEThSTeTH 1k1%18 1k) [|Yroeapn::5133616 7144) = 1i | f | i JA | o[rinsin ee ak oe en em aw 7k iimimete 1b ik tee 1y h tk Rh fA " 1 Ton: 135 (7.168) 1 |meme \ -- ih ik iw ak ish am aw Th Thim 1h re 7h im 1% 18 | Geol. EEEAN Wed oc 16 32:43:56 3000 fae 2 Primedica-Worcester Project number: PABX-BVA 413-018:PAGE 1-74 Page 73 Peak Area Report nme WO uno er s167 ov itameos1071 0 insettr: 7 cmm3eis cowmipsile Dowtoa wmo Emawm= Seme DivnsTee 101020001205 eae Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-75 Pages Quantitacion Report (GT Rovieved) @ HSSacaaFoFile |EDBEmE DDApTAE \BAS\PARK-BUR\L-062-1\OBZIOLLDSRoomeerr:aVtioatls:1t1a1n a 15 IacEeoErRacEion B params: RTR EWNiT.oDos\oE ass_sva. (878 nceseator) Reem Re ] { A | oje= IIi BP=orE m yr E EEEE EEEHAE N Laan 30% |!| A |i eo JA TEE 3 I. net sr : 105t r % am ew em= ax i th 1e6 niem 7% 7a6mTw Tn ] TT h w3 || A\ i| f rem i in e a Te Te ve e eTte orton maa wes oct 18 ned 2000 nage 2 PrimediaWorcester Proje number: PABX-BVA 418-018:PAGE 1-76 Pages Peak AresReport o Sumatera: 10C LOWWat napus: TE wee BIE rr Lo omp1unt Co=mp.re 2-t2am cage wmpapas rs Primedica-WorcesterProject umber PABX-BVA 418-018:PA1G-7E7 Page 76 Guascication fegort (Gx Reviewed) tSEaEtaNopirsotDBHo\SeED ADAT PAaB\eBRSKBURNS 062 S1\0E6231.D_perriitt:atl ]t33a a15 ETarTeocAasionBSuRsaEns: BTEENEE.2cos one svn (178 Tncesrator) p f= V 1 | = i E3e |iIh E =ee |E R EEE EEEERT TTIA TR TR E TRTE TTTER= T | E weap: E . 1 | I\ ! I. ees e+ mmm em mem mel em sree svete emer me i @LseoeSexepnt:akn2a0sa36koreeknow ahEoEk am Th ie Th 1 Th TedTeh e im ih e ih 1h 11 i i ih JAN | Te shieR Ta Te T Te ve he o6ri0iaD pAAN ved oot 18 12:63:36 2000 [I Primedica-Worcester Project number: PABX-BVA. 413-018:PAGE 1-78 Page 77 Peak Area Report Syme:tc oners1 rumenubDteFFeda:,m:0P2C150P.3CA0ATAPADPABCOVNRE seomveen anme 11357 Somgear Commons TReonptohn wm Eww eet J -- Prune 008200012.65 rages Primedica-Worcester Project number: PABX-BVA 418-0I8PAGE 179 Pages Quantitation Report (G7 Reviewed) PBDY aetaaonFiR le 1DE 1:5\'M0SsDe_-D2A0T0A0\GA1B7\:4P6ARX-BVA\-062-1\0621013.0OE peraVtiaoS lr:; u13a 4a5 nTosEegReaatiTMon PDIaramnse:iaRttE\NTW?ETA6O28D%BSV\A. (RTE Tacegrator) fo ~ wm [ " "T]| | | | wl Iih I | i i[J e ionns:m3vF33arrmITAT TR E R|eer | 1| | i i JAY ireieasas 0) weep: ars { I a ak th 1=TTT eaTR he th Bh1 . | i [I -- eA a (5% ok ie ussin sissw 7h Tw 1m 7% 16 rm vm vm ve im 0621013.0 PRBCOVAM Wed Oct 18 12:43:56 2000 wage 2 Primedica-Worcester Project umber: PABX-BVA 413.018:P1A-G8E0 Page 9 PaAresaRepkort [Isae menkccse tan onSsares 16 SSE Semcon, nA Com3put Confmpoitams Iswloa n Paomm tesme Prmedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-81 Page to Guansication Repore (07 Reviewed) PDYacaoR pirleeE : DB:\NRSD_DEATA\PAS\PRSK.SYA\A-062-1\OE21016.SDSepeercVtioarl! fiiatEn ae 4s IntEearEatiTon DFhucumsE:iawsT\EIE8oosPAS VA. (RTS Tacegrscor) Et it i : i eneefm:oo raaanrnR ----R ----R--TT) tSTIR -- i |! a pl T on y Te rr TT are a kepol @ saan: Bhooe pe) t | ER Ee Grrioies TEXAN Wed Ost i 12:48: 2000 [I Primedica-WorcesterProject number: PABX-BVA 418-018:PAGE 1-82 Page 81 Peak Area Report rote OCD one 1 135 nsrmestna Vome ACGApsopAeXBvAr6a armen 5 me3st cownmipstten Inmgaomgy tema tee rtngeTe 10182000200 fase Primedica:Worcester Project number: PABX-BVA 418-018:PAGE 1-83 Page 2 Guantitacion Repore (OF Reviewed) PYpSaotanRtPiEle+ DB :\ISDE _DATTaRe\PBURAB-06\2-2E\08A236B15.K0_GmpeerraVtioarl,:iw15a G48aInntWeghzoataion5sarRanCs:HNErFEA.ODS\PASBVKA _M (RTE Tncegzator) ES | wl IA | p mseSesomns8:15e 31G303128 36X36 68 68 &% 60 6% 7R% Tewne1m ) TR 0T5e 1% 7% % 7h Th | | - J\ i Ti aR aR Th TE 4 ge Te o Resp 930 T ee | || RinE va Ta ve ense sLkE dIiTe A7r8 TE 7TmiTr e Tm vy e te 060150 BARCHAN Wed Oct 18 12:44: 2000 sage 2 Primedica-Worcester Project number:PABX-BVA 418-018:PAGE 1:84 Page 3 Peak Area Report Samp: mr ome: nc nr SE UEShona oe mn eis congedt | Commpanns fa Ime mosmami bpm inionar PATSEVA 418-018:PAGE I-85 pass sn heLATcE omssciR on sertE(0 sevLiios1 oia | TE re E Te treT nton R pam:Tgrmewn. tr svesasen Fe treeeeeee x \\ : [p = nroRE G T reg | i | PoWRT ; ee ; \ | Tn shosk tm 1h im 1% 1a 7H 1k 1% 1h TE | es rem sd ar ER aR ER REE rimeticWorcs Project umber: PABX-BVA 4IS0ISPAGE 186 Pages [---- LJ om o(rimy nA s ..s.r Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-87 Page $6 Guancitation Report (GT Reviewed) [YaBHDlaotlanEtFilIe +|LD3:3\0CSehDeeDBrA0T0e0R\aPesArBmi\nPABK.OVA0V612 1\GEIOIT.DBE oerVsiialE t:: t17a 4a5rstotSesErEaRtiAo"n pBacRenSs:HmGSTEFNFEADGDS\PBAVSA.K (RTE Tncegtacor) =pu it { i pe RST wasp: I5R97 TTS || Osoas p: 160doee ARGB TgTle inTiTh th . -- if\ | i jLeLT | Te re ai tk ik Ute hm a veh skem Tm 7% TE 7%14 1% te tn ik rh Geai000.0 EBX UAM We occ 18 12:45:13 2000 rage 2 Primedica- Worcester Project number: PABX-BVA 418-018:PAGE 1-88 Page 7 Peak Area Report PySue mit:220e 6 ic OnSems a017701024. nmrE bhFr aPnoe. ADMEaCAATAPAGPAVAKL G62, mamma 1 Gomete Commun ISL m tem mest Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-89 Page 88 Quantitation Repost (QT Reviewed) Qo RDeagtaonFi|le:: D1i:7F\U0SGcDC_D2H0IA0GT0HA\WPaA29tB:e\2rP4ABK-BVA\1-062-1\0621018.`0fOperoaVtioarlr:: ew1a G45usaIcntMeagtrhaotdio:n DPair\aHmPsC:HERMT\E1I\NMTS.TDHODS\PABK_BVA.M (RTE Integrator) a Se Se =o \ | - \ | ben iL ToSi T Ee ---- eo w 08 TfR-- oIB TE Te7EET-- RE |=Son: 7133330 (73407 pe isso RSeospn:: 153554.5 171367) A Th ie Te Te thiki] Re -- A i = -- SE I VE | Len oe dwoh eh 6h sk om 16 7h tn 1 3k 1 70 1%101% 06210180 PABKBAM Wed Oct 18 12:45:33 2000 vage 2 Primedica-WorcesterProject number: PABX-BVA 418-018:PAGE 190 Page 89 Peak Area Report [SeSacmcplhei: 2t10oCMdD] oecnaeet:ern 0.43 vn EASE ECBO RVR Gommts Commons IEMA gn hmmm Smet 418-018:PAGE 1-91 Primedica-Worcester Project number: PABX-BVA Page 90 Guanisation Regor (G7 Reviewed: @:i DBE artaatFitR o DB :\R WSDa DNTANePASi\PABK.BVAVE-062-1\0623039.0_Dmpeernviianll:l S19e a45 lTotEegzRatiTon B furans: A wnEEn.oDR ns nmE e_sva. (0 savesracor) ------ ---- PE' bxeeSeaee pm+:i5 3085V 7t .15E 71 C I = ETXiR ne NL TR 2CTER TR = 3 ] | IE Cp Odhci soe raa m | le | Cao ieoeTa TeeT Te 062019. PEAR Wed et 10 32:45:52 2000 nage 2 rimedicWorcester Prot number: PABX-BVA Peak Ara Roport |J 418-018:PAGE 1-92 Fagen Sma Primedica-Worcestr Project umber: PABX-BVA 418-018:PAGE 1-93 Page? Quantitation Regore (QT Reviewed) @ Doi atnaePile +|E D1:5\WGSaDe_aD0e A0T0A.\PR2E0\:P0A3RK-BVA\L-062-1\06Z1020.SDEparavT ciearl;:Sw20y wee 45. IntEegnraetionBPahranaa: nsEENIETF.HDODS\PAB SVAN (RTE Integrator) =o i | - { oeSroe na3-- S-- I ER ST wai 767 TSeToeRmTTSRT TR 2e08r T TEe TR ee T | | f i o| iReonngt: e133s8i0e00a.k16se e>ek ihTi T e TH Bn 7% TeT re Tb i { - _ A on in eo e oe ea skaete th be re ier Th Tm 1h = | ih tk Ge2i020.0 PASK_BUAM Wed Gut 18 12:46:12 2000 sage 2 PimeticaWorse Projet nomber: PABXBVA 418-018:PAGE1-94 Page Presk Report spre co oe ron en EEE semana, mnEE Primedica-Worcester Project number: PABX-BVA 4I8.018PAGE 195 Pages Guncitasion seport (OF Reviewed) ozDfBatoEamSFile DLo :\SD OCNETa A\oPoAS\wPi eAeBrK-OVR\A-062-110671010GmpeeerraVtioarl:: i2H1a 1f5 bsacesration i arena: ETE.A -- gr Sl | | or weeepe: G3R0r rRaO Ch Vk4 TeiT . Te Te 1 Te -) emer mmm be mere J ET TE Re QneLon: 3s oe a) || f Gin ie va shiibin a To 7%TH 78 va ve Ta7h TB oeaion mmeAM Wed occ 16 12:46 2000 rage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-96 Page 95 PeakArea Report o NSoumep tsame:3C165oerc Poa OweaCesti1n 0 2002 Compass Gommogums THD gy Pm temo Primedica-Worcester Project number: PABX-BVA 418.018:PAGE 197 Page96 Quantitation Report (QF Reviewed) @ DBiaetaattonFile : D1:5\E GWeSeD-_aDE oNoTnA\PL AaBbNeSrABK-BVA\L-052-1\021022.ODfpe_rraVtioarla:, ]2x2 we a46 AsotSegEeEaRc"ion BPRazE ams: Ro TENT.PnpAs YAM (RTE Tategracor) pe il nR esoorsi:oisaesT 3s1 rr Ce hGe S lp .Tio i ovee remit 5 osen F:ont33s08tren | tg, Ere | mat IE ee Re" | it TeI eT A TORR! ! EER T eh ERma OE2022.0 BABCEAM Wes Oct 18 12:86:51 2000 rage 2 PrimediaWorcester Proje number PABX.BVA 418-018:PAGE 1-98 Page 97 Peak Ara Report @ .TERTEERET ac mnLE ESSLsma rosn. wandR Primedica-WPororjcecetnsutmbeer:PABX-BVA 418-018:PAGE 1-99 Page 98 Guancication Report (GF Reviewed) SAattoan"File D5P\WOSbRDs_eD-AaTsAn\mPA2iBn\iePoAnBK-BVA\L-062-1\0621023.S0E peraVtioaS rl,: 2w3a @ ux 4a5 nIncSeaTrIaRt]son pB arams:eRTEDah N.HEoosa \eABK_n SVA.M (R78 Integrator) sn; em, EOE HReeop:r3n280 ase Ta TeeR eee eRih romeeseeny JA | @ fShaonT svessois de e eh at k wl Teae ,.TH Te TaR vm vm teE te 1m i i i | Gon ea i ie wT Fe 7 7 TE TeTe TR Te tw Gh2101D BABCEVAM Wed Oct 20 12:47:10 2000 sage 2 rimstica Worcester Project mmber: PABX.BVA 4IB0IEPAGE 110 Pages [p---- o SE SampleName: 2xGCLOWPrasma OT DateAcquired: 101720002121 TIS mss nS Primedica-Worcester Project number: PABX-BVA. 418-018:PAGE 1-101 Page 100 Quancitacion keport (GT Reviewed) oDi ataorFile |T Dh:\WED ORENTAE\BAB\BTAKE-BVA\1-062-110621024.0STpecVeisail:; i24m 1f5atTeacegtacion frarac: wiTA . 8 p -- sory - ) \ [+SSe oh a SRi on Tio Te inTh Te The 7% eT Ll aTg6RT131 (7.134) TT | --1 A | \ | e RCC n C e ar leeL g N S T na oip noesp 1332 -- - moire" | f----i Dina 663% vian bis Tm tis rk 7% Tes 1s 7h Th Th. GeioeD PREXSUAM Wed Oct 18 12:47:30 2000 sage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-102 Page 101 Peak Area Report [IcSamcplleo:t CrvotwPeema oeATm EESpm NL, Onepcaree e0172002110 Gomm2ose cwonimlpdlans LaTelemmo tadlbam iota Primedica-Worcestr Project number: PABX-BVA 418-018:PAGE1-103 Page 102 Quantitation Report (QT Reviewed) @ DipaoaptePile:DI y : MSDG r DATA\PdAiBNvnEyABK-BVA1-062-1\0621025.0Ge pecaVtioaT rl:: a25x G#5ianttotWeEgirhaotdionTb parRaPmOsH:EIRATLEINNETT.H2ODS\PASXSVAN (RTE Integrator) FE EE pt we Eb 0Tg iei TT R ET Te Ai Fsleaom. m35rasg EE emcees) i i | oN aE QfLrei a ET TTE hi eT Th ts A . --| i | ai : i ie eekeen ake ie eb Ta TR ve 7 _| TeTh kia oGa1025.0 PAE BAN Wed Oct 18 12:47:48 2000 rage 2 imate Worcs Project number: PABXBVA 418-018:PAGE 1-104 Page 105 Peoksmnsap |J st nae oneEG evmans, wdTES Primedica-Worcester Project number: PABX-BVA 4I8.018PAGE 1-105 Page 104 Quincication feport (07 Reviewed) DRBaotarEFileE{9D:\WE5D ROGRTeAt\onBAneSNaS"tATeBatKre.rBYA\S-062-1\0G21026.0Sep_ekrvtioarrl:: 26 [ Ea #a5 rtacVegErRatAson pBareamhs:atEyTEDmNTiDmosa_ovA. (RTE Taceeator) Rg mm + pe ? \ -- | I efiseoooanmpii.:rshE ist s =T TEEL) sTe TSEE T Te TR =1 i ! es IA i| QReCI aopn:t s 42E 46 re y eie EE Lin TE Ve ae ee mis 76Th 78Te ra iaTe Th ER G6EN026.5 TECWAN Wed ook 18 12:96:08 2000 rage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-106 Page 105 Peak Area Report NotboSnakmRpoearme:P32A01B0BWaAte AcSae:m 7000210 ruCem edraame .ABenhACATAPAIBAB VALE, tment Sonm3adt Cwomimpistens InWf)omfn BAaNEE Self Primedica-Worcester Projct number: PABX-BVA 418-018:PAGE1-107 Page 106 Gunication feport (G7 Reviewed) DfMatoaEtFaiNlneiA DClio\ESaDDNriTtAig\tPAei Br\PABK-BVA\E-062-0623027.0SmperaVeiioarl:s 2a7 @ nist : f45 rTocegeation t Facams:tSTEIN. 3 Ap] fo sna, ) i h i J 00Re 66 08 oi cw en sm sw Tm Ts isTE ve a 1w 3m Te Tw | S Resp: 2059emone | @if SlfxosntTi nsssee 5% oe 100 em 8k TTEh 1a 1m 1%" Te 1% 18 LT oh i\ | PrrT TT TT TT i 0620.0 MBEAN ed oct 18 12:48:28 2000 sage 2 PrimediaWorcester Prot mabe: PABC-BVA 4IS018PAGE 1108 Page 107 PesRpt Son RsrRi s EToE SwCio zn EI sn mon REE PrimediaWorcester Project number: PABX-BVA 418.018PAGE 1-109 Page 108 Guantisacion Report (QT Reviewed) @ HwDataFeorPille I+1 5H:0 EEDATAPANB\YPARK-BVA\E-062-1\062302.DS_EpersvRiiaalr: E2i0m a#5 nTocTegEeaRcionDpeicanas: nsWmheos passa. (R78 xacegocor) pon ra -- Pe i i t{opbeeterTrEemEA orse e ever aT h pT TeeT r oSeTegeee 7TR Te=| Pnorlm: s336 am =| Ea A i TnikTe TET ye TaTe Th Oertp: r $310 r m.m BEER! ee JIa i 04STeiV Gdn7Ta 7Te1seeTh re Te GGRi02.D TREKBAN wed oct 18 12:48148 2000 wage 3 PrimeicWares Projct umber: PABXBVA 418-018:PAGE 1-110 Page 109 PevesaRepkort mim EE nnEEREE onorian mnRE copes comes SES wm mame one rimedisWorcester Project umber PAB BVA 418-018:PAGEI-111 Page t10 pce Fite DASE DATA ABowPne.itUatRiLon06Report G7\OGZIB.D_Revakise) 32 oTEe ome Saree tay J EEE CRE--been a (78 tnt) -- I I | Ee [Ioar NPR TT TI N pe E TeWET T oiital oie Tow n wm ek Te EERE ThTE 7% 7% 1% 1h 1m re iw | II | TE eh te ek te an om 6 76 Tw 18 7% 16 1h 18 1% 7m 7% Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-112 Page 111 Peak Area Report Simms Some a menCtEbein aa APBSACATAPABRABKVAIO Inte fmpolt Gmmpie IStMs) gm heAmE beeie PrimediaWorcester Projet namber: PABXCBVA AIS0ISPAGE 1113 Page 112 aeseitarion hepore (05 Reviewed ofsi ratefPiie | UE GuDmDE pATmA A\BB BUNVS-062 OGDDsporeeresviinas'l a30y 15 sncEearRation P parame: TsEeTee ona van (v7 Sosearacor) org re -- -_ El 1 - | Leo EEE oR TR TEETe Peer | wRios GBLh TFORT i | | ree SEE ER ER ER | Resp: S002 . i cm ee IiA || 4B 5% 6% 6% 6 SN G06%I 10 1m 1% Te 1 10 70 10 iw | GGrIOI0.0 TRIAL Hed Ont 10 12149126 2000 ne 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-114 Page 113 FP eT ree orEia JO TTRIpaE s wer EET 7 = il a Primedica-Worcester Project umber: PABX-BVA 4180I8:PA1-G11E5 Page 114 Guansitacion Report (QT Reviewed) @ DemataahnFile +DDt:Iy\WESDDaATA\PnAhRe\PyARK.RVA\L-062-1\0621031.DSaperraVciearl!s w31a 4G5an1tacWeigeehaet"ion DpFa\rHamRsC:HEKNT]E\IWNETFDHOD9\PABK_BVA.M (KTS Inegrator) ae --y - ih oes 6%4CR eh G tho 4% MRoansp:: 2s4385xwrm G8 $7Taie10 Th TR 7% 18 TH 18 18=:| GI ERE EER PR GR kPeraigs: o43s55 sen res| re i . I5.SU ---- ET RTT RT | 06210300 mEXEAN Wed Oct 1 12:49:46 2000 rage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-16 Page 115 Peak Area Report NetSboaokmRienmee:: S3O1LoA3Sn6m OeAS cquismon7200 257 Gomspudt Commpilne Lege) gm Sm Peles `Primedica-WorcesterProject number: PABX-BVA 418-018:PAGE I-117 Page 116 Quantitation Report (QF Reviewad) tDSSaetcEatoNrFile:| D1V:3\\CoMcSEeD_DSt0Ao0Tw0A.\mPaAaaBta\ePrABX-BVA\1-062-1\0621032.S0EpeEcaWcaeirl!: w32x l15anskocWeEgezhaocdi"oTn DpaIc\aRssR:CHwErTWeEnO.SS\PAR BVA (KTS Inegracor) J memmmmenomm Tem ft of it } pp otVe nfsGa nketmTeiiS TR pT eR er o wepm :Ioo: n (.A54 . TT | OL Re Re G-- G ds~ Te TR NN e TL TeTe--i_s sh e_in worm: soso" ieJ TR A{ i aa ww me Te ve versie ve TR re | 062032. PALER wed Oct 18 12:50:05 2000 fase 2 Primedica-Worcestr Project number: PABX-BVA 418-018:PAGE I-118 Page 117 PeAreaaRepkort @ Loam See media reser Fj number PABXCBVA 418-018:PAGE 1-119 Pets mon spre 320 esos ouHt EIR iy C LS T B DREEooemassn ae eens Md TT me | | I | Tees 46% 0 6B ow st am Gkaw Gm 1% Tw im Th Te 1% TE TR 7E TE i I oh IEEE aynA Lie om en ak elTm Tw = (7 3 is3 Enea Tn en sw ak skeh em am im im 7B 7% 76 1% te 1% im 1%| Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-120 Page 119 Peak Area Report Notrree PADS Oe Gomi2pats Comlpoantans Tewpeol mfn hima haope Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-121 Page 120 Quantitation Report (GF Roviewed) DBSaetEgaoNnPile:"iD:T6\OW0SRoDeD-H2A0TG0R0\PeA0Be0\r:PA3BK.BVRL-0G2-3\06ZI034 DGSep_reecraVtioarl.: W34n @ WE #a5 rsatReRoEsRaSiAoTn DPazerns:iseTEWNE.pmEoospax ava. (RIS Integrator) - i4 | hoe oOTR TT TT ihe fei fiM iResp:a4820 onae)es | erm EY SOO em JA ee] tTons 135 Te (7.162) Ta Ti E Te ThTeRi FTa " neup: 3500 ji i i | Lisieie aio wb enam eR TeTe Th 1% se ie re th vm TR | GEa10MLD PABXBVAM Wed Oct 16 12:50:48 2000 rage 2 PrimediaWorcester Proje umber: PABX.BVA 418-018:PAGE 1-122 Page121 Peskea Begs eeJ nSI SEBE ISE Rcovamassncr, w-- ST d me o `SampleName: 4 GCMO Water DateAcquired: 10182000055. pont mgs SE ag mee rimsWorcsstrPictumber: PAB BVA 418-018:PAGE 1-123 pager2 JBainrise I+ SumIEmReTe QT LRurllE t Be F E ro ereEi seron D pl ees si rsR neo ar n1 SsgerenSR po i pi fi Ti n Ts 030G0SH 6 564k A OR 4% [- AN 7 gw TE 7% Te Th 7& 7% TR 7B 3 room Lhtee sm a e an k EL LL sk Te Th th 1k yeTe ve 7k te 1% 7% Tm | I OR en eT en ee aw Te Tho ve 1h Te 1 ik To tk te | primed Worcs Pot umber: PABX.BVA 418-018:PAGE 1-124 ge 123 PokeReport SeR eE e srnprmmsponis oeR ERENE en oS er7 r m=peHgEmEam pp Primedica-Worceste Project number: PABX-BVA 418-018:PAGE 1-125 Page 124 Quantitation keport (QT Reviewsa) @iRDaocanFide :E D{E:G\EHRSDE _DATAP\YPAB\BABK-BV1A0\6A21-003662.-Bfp_erraVtiosria:, t3m6an RE a45 nsatFesAcRat"ion B vasans: i KTEOT.hPe opsphsn _wva (578 Integrator) rg ea 0 A |- il | pFr EReeE sp:n9R s35 er Ces r RR T i ST s my i it fem = i sn te oe ene 502 TE O | nBeSop: 9E 60 iE Sh ak ve Te ETis Rth 1E4e R18 Th TTEh tRh 1 || AA lomo ; JAN | SE Te i we ae ak im Te Te th 1% ve 7% ve Thm 16 0106.0 mESAN Hed oct 36 2151:22 2000 rage 2 Primedica-WorcesterProject number: PABX-BVA 418-018:PAGE 1-126 Page 125 Peak Area Report NSomtetee 3A 05 A 00ie ecrum: 10200013 ne oSsAmiTuEm 0CS 6U2037u 0 esinss 51 mme enotn rei: 75 Compass comple Iai) Bn mami pene Primedica-Worceser Project umber: PABX-BVA 4I8.0I8PAGE 11127 Page 126 Quntstaion Report (@F Reviewed) tDRSaetlaoenFile:|DI3:NRRDDETNATRS\?wAaBtR\eBYrARKBVA 062\0821037.0_SFpeErsviioarl.s i37n #S5hanIentFegarhaetaionracHamsesH:ansE.EpFHPAOS SDVA\N (RTS Tncegeacor) = ey wewefoSr E (v am i e --TRTNJR , iiA TT TR-- TR. TT Te-- naam: 308 ) | @neem: EeTeARTs Te iy a he EE soi mee RR| eR - A i Cie ea sie en ib ih TaTeTe os ih TeTe TeTe ie 060.0 PRE EAM Wed oct 18 2is1142 2000 nage 2 rimedicsWorsesie Pret number: PABX-BVA 418-018:PAGE 1-128 Page 127 PeakAva Raper LJ a onEE maa, mSen EES comet rz -- Primedica-Worcester Project umber: PABX-BVA 418-018:PAGE 1-129 Page 126 quantization Report (GT Reviewed) @ i uafrcmisaeoePeileDD:o\MEmEDCEpATAAVPa AB\PABK-BYAV1-062-2\GGRAO8.0SSep_zeerr:vviianlt: a38a 45 TncResrRationSsaAganS: aTEWIaNi.8oos\eanvaa.x (R72 Sategiacor) - hy |niep: 81331% 7.2330 )iE 4 || | Te TE eT ge ee | ow fon: 1 EU - )' 1| ih i Ie eii ae ee TR TeTa i wean mena ves tut 30 sees 20 fae 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-130 Page 129 Peak Area Report Sameer 0Dpms osc ren1s met TR UESCATRADPASLOVAL ELT, rR empeds cmmues label im egw hme Primedica WorcesterProject number: PABX-BVA 418-018:PAGE I-131 Page 130 = | cuancitation report (27 Reviewed) DfE aotrageiR le DR p:\ED DsATA\a EAB\PAeSKi-BUA\L-0GE-\08R103S.D_SgpeorVerioadre:' t33an @ Hi : J 45 nvesE racion HFararss: sEDIT.D AE -- Rr g e RR1 e |= JA { pT p E ET a Tg 7 7 o= is rar i == JI i Oi nLiin: i ayaia ef s8T : aa | 1. EETe IN i ww mT Te Te Th ie rae] Geaion.0 SAMA Wed Oct 18 13:52:30 2000 rage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-132 Page 131 Peak Area Report . OQ Sm ar ESEE Brenan, mn REE Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-133 Page132 Cuascication Repors ar Reviewed) BBE aecartFT ile 1 oE Sr\SD BuAEmHayPAAB BRURNS 062-10621040.0_SroennciVeinntd!: ti0au @ iE E 15 IovT earacion seratne:aIoTns seovna. (x7E Incessator) a -- ST -: iA | p-i I| | f bowsiee no:lai6s5s75 Te 6% twGmsekon6 7Bo E ) S in 7 .RIE | h TE-- im | | @U aF!irinest: HsoesreesTeES rinetahs anh kik i rhi Arh Tw7TTSeTETh Th 1--m =i I i n S in akiE sh she th ew sieiile TteehTE1%T1 TTEh T1hETTh E tE e| orzionn. TASKSAN Wed oct 38 12152040 2000 rage 2 Primed Worcester Pret number: PABXBVA 418-018:PAGE I-134 Page 133 PeAveaRepkort LJ SST ne SEE EU Sarena. mnEEE 7 Wa RTA rimedica-WorcesterProject number PABX-BVA 418-018:PAGE I-135 Page 134 cvneicasion fepors (08 views) @ f MisauocmFail r SSDLONTe A\PwASiVPlARK.SURV 062-1\OGHOILD_Sapees:rtVitaS:h B81n v5 smeA egracion E Eagan ym.ol on sna so ans Taceszaor) sr em------ , -- I | 1 - a res sla 04% 0% ssh GR ak 4k 1T% h Tk TR TH 7% 78 7% 18 1% TBrre Tefil | we - ---- AMOUB:E DO ee semen rpg a.- 1 L Yo ser a = Ee i, | TE ow Gesh ie eh em am TR wm 7% ie Th rw 1% tm 1s | |reaps 4850 . 1 | A i TR rRp a TT[i Te 1aensaeTei8)i Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-136 Page 135 Peak Area Report NomaSumRiaeme:1 FAi S06 OnpSqsm0820023 Iosauppodmaamee:SPHrAECEtBVCAATAPABOPVAABLC mtrVa omp3elt Cowmigpslim lemoel faam Se : Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-137 Page 136 Quancitation Repore (07 Reviewed) @ fDiaotartaPile N tDy:\e MSOE _Dt ATA\R Ge AB\PARK-BVA\-062-1\0621082.S0FeerraVtioal:: M&2ax wee e45 nTncBegErAaScTion EPaIraRmsS:hutRAT:E(INET.2THAOBSDBVSA (RTE Incegracor) --_---- | -- remany\ : wu i | i I re a rasp: 737 | : --| |{ ifi i be JIN J| CR RCWRTTi o| ry oomnpt 15286 (77e 60) o i Amount: 80 R e re i | fi\ i ee JES IJ N-- "| Fwmae we Te TR ee Te eh Te Te Ce0s2.0 PAEXEVAN Wed Oct 10 12:53:18 2000 tage 2 pimsticaWorcesterPrt mbar: PABXBVA 418-018:PAGE 1-138 Page 137 Pesk Area Report [] wnSEER SampleName: 5 xGCWO Wala: Sp `DateAcquired: 10/182000 331 J OT .. Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-139 Page 138 Quancitacion Report. (QF Reviewed) @:i DoSabtlaeenFile D5\WGSD_DwATbR\aPcABet\rEeASK-BVA\I-062-3\0621043.B0E pecaVtioaS rl,: $t2a G45ianstncWaagkzhaotaiTon DraIzAamRaR:HwEArWeEnFsAGO\ART BVA. (RTE Integrator) forte rr Ry i I eo roTaonn:: B131h (7,230 UR AB RT TeTE7 7% 7 7 TR TR | | I [ L[Ptenatta omAiTs etrdhenseen emiT m rh ve 7h TT h 78 Th vm im 7m | ET----e-- e a--a--i--e---- wn--Baie Late_-- e 7 ge s TET TR | Ge00.0 mex AN Wes 0st 18 12:53:38 2000 mage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-140 Page 139 PeakArea Report @ mmm ha i Core comptes IM wm same ype Primedica-Worcestr Project number: PABX-BVA 418-018:PAGE I-141 Page 140 Quantitation Report (QT Roviewsd) tSoDapcheaenFile D + Dy : NSe DDoAE TnAa\wPieArBra\rPyABX-BVA\E-062-1\0621048.0Caoezavcioarl:: $4 4S5aIrncSegErEaRtsAo"n pD arams:IRTER DaTr.h2ooS s\PA] S ava. (RTS Toteszator) er men ---- ia I! ! R hP owe p5tr TteEentn iT AmountT T: eTR B.e00rTR a Te Resp: 35 i i iI Ph r|S w ih tiemeenaA T aTT ih n1sI 7 sia i 1Thh, | nafconn:t 143s0 1.260) A i i i | TeAnE vei wm ww ih Te Te Te ie ve re in ie Te 0621000.5 BAEK BAN Wed Oct 18 12:53:87 2000 sage 2 PrimediaWorcs Projet number: PABXBVA 4ISOISPAGE 1142 Page 141 Pas AraRept O mm fac Primedica-WorcesterProject number: PABX-BVA 418-018:PAGE 1-143 Page 142 uitaion Repore (QF Reviewed) SEatEaSorPile +1LDR :EVSEDR DfNTeAd\PeAtSTe\rDRAK-BVAVE-062-\021045.0_SEeerbViiaanl!s t5an @ ET 45 socEegrAatiTon EPaPzanSs:iotFEWTEEFDoo sham. (R72. Integrator) Sret a oy,i | oo - i i i L F eT eA tE ge R emeR en] mMLsoenep:! 13341 0.1201 i. ae i"Amount: 6.00 on E-- i! | ni in -- A QTaE iie ee ek esko en h amR ew THTi TRETTH 1%eTE 1e 8 1% tw 1 naan i | } i | Lin e546 8% em 6% eR 6 T@ 70 1H 1% 14 1% 78 70 18 1% | Geos. TEXAN Wed cs 0 12:54:17 2000 wage 3 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-144 Page 143 Peak Area Report NotoSboaomteaNnacmee:A515 VA10p0 57s te Amide: 682000420 nen SEL EUS CAOASP OAA L, mm fomgets | cmgpttem Ima beam tame Pied Vestsrs ae FAT A 418-018:PAGE 1-145 eras nein vs. vt o BREE et iy FR -- pg TI - I ! mes 670 4% SH 4k 4k GB Gh oh 0 Th Tw im 1% Te 78 78 1% 18 1% | o AASi : : =| A TT rrr a a Se rer = ik se se ww se wn um sk10701% 1% re re re im im 1@ | Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-146 Page 145 Peak Area Report NottSambprle.:C30O0ROis ue ASnam018000449 aminesOeuaptotdirmaaapnmae:.GScAAcAeSrBCAATASAGEADLEUAOE1 mumreatneo 47 Gomg2pats Coimggetuns leool mn faREp ees Primedics-WorseProject number PABX-BVA 418-018:PAGE 1-147 Page 146 uaseisacion Repors (G7 Reviewed) @ iE SBeatree]PieE : D:R\NR SaD DmAa TAVPBVRAVER-06\2. E\GGRZIBONRT8SSppe_arr:Vstieat: i47hn 1 satEegraTcion pSaramsE : ETEcE2osshss_svn 478 Tatagracor i a,] -: A i BA ERr EGEN p a Ll ) RAT a | i NI | R_---- E a--_-- [= neEoant: 8ss3s3 eo = EA RAARER EE R Amount; 0.00. -| JiA | Ea a Re Orono EXAM ed oct 18 22:55 2000 noe 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1148 Page 147 Poak Area Report NosSeamRpenleee:P20A5DLOowVp0Ar8s oe sSesn0102500308 ven UE Snipes aa mA Gomes ope Saba) bn beam Primedica-Worcester Projct number: PABX-BVA 418-018:PAGE 1-149 Page 148 Quncication Report (QT Reviewed) SDBaehttaiotnFile:D:130\WBSeDrDAElToanA\aGAaBN\nPERRX-BUAVI-062-3\0621048.0Sm oeraVtionrS k:: 8h8ax [ eal a45nTtaceeghraoc"io)n DpTariaRmCeH:aMBTSEWLET.FEHODS\PASEBVA (RTS Incegeator) oe BR -- oo showy semen ol A L p Preoe a ose e NRa E EWR PeNeTr TT reYny [ret ae iA i . iA | @i| aeEnep: E335T0 ei .i ih i ee TeRahnts hs6.0a0 re th tek-- | | i i | i i Lb oE e dee aie ani TeLieE AveTee vea ve eve teeTve || Geaiow.o EAEKSAN wed Oct 19 12158038 2000 page 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-150 Page 149 PeakArea Report o Sarge: Cmts oetei risa Sort coe SR cnt copy Primedica-WorcesterProject umber PABX-BVA 418-018:PAGE 1-151 Page 150 quantitation Report (OF Reviewed) DfLataEoFilee :EDVCDDGNTA\PAnSa\eDeABK-BVA\E-062-1\0621089.0B_ToerakViioaetl!: t$9an [ ea 45 xntEegRraAtsTon Sseciam:aTEEmoEns pha sa. (RTS Integrator) pram em" wl A | Pe jt | \ fohese ITE TO eeecm!TR eR TeTR TRE Creepn: T35H6TRIN So | TT 1 f | ema oTrh iw .sw ew nism: as | Corere Ned me | ee em ek ek 7h TRWeiTnRe 7% S7% a7 | ie 1% ik im | } Ji |i ei es JAN eve 7B eeBeAT RRRT)| 0621009.0 BBXSVAM Wed Oct 1 12:55:34 2000 rage 2 rimedisWorcese Project number: PABX.BVA 418-018:PAGE 1-152 p-- [-- Ja SumoteHare: 8 1GCHGHViole rE DeAcquired: 10182000547 won SEcss, web Prmedica-Worcese Project number: PABXBVA 418-018:PAGE 1-153 Page 152 cuancication sepors (F Reviewed) BDBeuecsaOrilpe:iLde IDO:DI DATA\waPceAtB\PVABAKS 062-1\0671050.0__Sopecrtiine:tt f5i0an [ Ev5aTncEegeaAcion pB aans: sTWeuEEre 8rhons\anax svn (x78 Tacestator) = er TF re | = i i. TT I TR i Bram:eime a S) Es =1 i O sG eeme : bsTer e a am en em ew 1m 5ae | JI\ 2) 5 wm --_-- imr e e |! Eee ow a ie er sh em im i 1H re iw ve iw tn 1 1m cenoso.0 maka Wes oc 16 12:55:52 2000 re 2 rimetica-Worcs Pret number: PABXBVA 4IS0ISPAGE 115s Page 153 PereaaRepkort SiE enEe wgmmpen e a I n g TE asso. n_T ---- corr sone SE wn sam capt Primedica-Worcester Project umber: PABX-BVA 4I8-018PAGE 1155 Page 154 unsication Repore (GT Reviewed) HsRaetEaoEnFile 1 DR:e\NSDi_OATA\aPnReSGs\PYABE UAVS -062-1A0621051.D_SEpoernvtioarl!: t5i3 @ uF 4T5 sEateRgracTion DparRassS: ETEIIrT.oEo sre wa. (373 Integrator) ef Do = +R a [ern Boke ! i ; i | ja | TiT TeT eoa 1 TT A fmBh a aaeT mRd TEENA a ai hE | onaal n Pe L|p-- | i i LF o ETE E | SEEREEE GeziosnD mm VAM ved oct 18 12:36:32 2000 nage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-156 Page 155 Poak Area Report o NoacSkumRiaanme:A5 OLowAva omsS equaasorucone 625 smeuhneaFsimnaes::S521AE0D5B,20AD.ATAPADEADCVAIO. nrt uima emnees573 ongpnit Commpines Ima ln Rem Spe retese me 10142001256 an Primdica-Worcester Projet number: PABX.BVA HSOISPAGE L157 Page 156 tn Tg 5 Swit oo pPaopoils DIY WESEa EETOA o OE TTOSE ,e9R llTh A A EE Te SEE wg= ---- i) -- | |- I L pree ew a ) JA TT Ta || Tes 10 $5 GB G8 Gh 4 6% 6H eR 18 ie TH 7% 7 TW Te1% TR 18%) A Th ew se ah sh Gn im ek 1h 1% rm iw 1s 1k 7h a m 1m Rep, S187 . 1 i i1 1| | TTTEETTEEa Ri TT EERTa EE Primedica-Worcester Project number: PABX-BVA 415018PAGE LISS Page 157 Penk eager mmm em wTo EE eames, wel REE Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-159 Page 158 Quaaicasion Report (GF Reviewed) @ieDSanicealnaFile :1D:Ee\xWoGGDe_nDuAsTgR\PvAaBet\kEeAeBX-BUA\L-062-1\0623053.0Ge p_eraVcioar rl:: w83a 1a5 nIncGegErRaIt"ionDPIarT ams: e RTSIENTTc .HPODSh BABYBn VAM (RTE Tntegracor) a necen, em ol 1I |- I I#mpes567065T Teaa a3 OAR oaRk 44 TTpeTh i1sT78SvTee TThiiw i1fh a1e8 18 Resp: n4a6r2prays i| i i i QR xaonnt: s13o5ds(7.163) |-- a -- 7 | Ii | EEEWe we oR ae ee 7Te in rh ve is i ik i vel 062105.0 PARK SVAN Wed Oct 18 12:56:51 2000 sage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-160 Page 159 Poak Area Report otSuampreasme 4A2B0BiRtp8ana mesSenums aenaznn705 ee ERE ELE Shrsmest on wr emi3gedr coimonpiims Imhplomfm team hips Frown Tn 008200257 rae Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-16] Page 160 Quantitation Report (G7 Reviewed) RDSaetaEaonN*Pile 1 DE:3\ISDNaODK0AeTrRa\HnPoAa0B\rBAa7B2Ke0-8BVA\1-062-1\0621054.0SFp_eerrtaVtioarls: w5ax @ wid : a45nsencRegEeEaCcEionDPIarUasM: RETEINTT.?HOPARDKBVSA.M (RTE Integrator) == i | | pEe Esen hvoman heah 4anTTeL i\ TeSTTRR Te RTGe Te Te TE Te TE | IRR mE nrop:T4406tro ee. 7 I | onnieenimt:ke123580l 6 (7s.1hs 59s) k en eH em 7m 7%oir1:B >1% . 70S00TH Tk TTR R7%E1EB a] f A i CIN i GE1105.0 BAB AVAM Wed Oct 18 12:57:10 2000 sage 2 Primedica-Worcester Project number: PABX-BVA 413-018:PAGE 1-162 Page 161 Poak Area Report o NetiSampRlensame:0 HER 0PaS5n5s opSusi0182000725 Fo A,SATAPBAUPN OA2X1 mammami 5 Seo3met Gmwsipsiun Img mooom taume ellen Primedica-Worcester Project number: PABX-BVA 418-018PAGE 1-163 Page 162 Quantitation Report (QF Reviewed) FYERSDae:tEcaoNnPile ED{:TaWR0EeDeDziAoToAn\mSAaR7r\:Pe3A8BK-BVA\L-062-1\0621085.SDSpheereavcioarl:: w55n a15tseotEegEeCaAt"ionDaIzaNns:rE1Ee WTET.HH ODS\m PABKA _SVA.M (KTS Integracor) [on amg | ] | [--. i{ { i| rele Th ew vw ew sl en om eR TRTe Preaop: s24t52oma . a wa } | fi | bee Te iha i i deem 1 TR Tie 7% TB ie 7h Tk TE O= l a E. S a-- : fi | i i i J || Ua ox seams oh ek ek Te TW TR 1% Te 1H 1 1m TB IE | O62a0s5.0 EABXBVANM Wed Oct 18 12:57:20 2000 page 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-164 Page 163 Peak Area Report NomsScaompRleenamae AGAL2o 008ms oe Aocese0na047 emnson masz opium loi team tah rimedica-Woreestr Project number; PABX-BVA 418-018:PAGE 1-165 Page 164 [I -- QiSfB a"tarPilae L | Do\WEDCAT TR\rEiAaS\BARRBURVE-06E-NGKELOSE.DSe.orereeViiaatl::12i56)y E1 saEcepsaAcion DFurRaneT : ETEE DIT.E LaaEava (78 Tocegracors fog vena ree |= i i - i | - ;A i pe ER ms a0 63OB 6 6% Sm sr am 6m TTww 1m 1% Te Tl TE 7% 1% 1% i | 5% io ah Gk eR sk ak Tm Tl TE TR Te TH 18 TR TR 1 i J. | En a% ek aio 4m ew 6 Tm 7% 1B 1% 76 TH 78 1% 2% 1m Primedica-Worcester Project number: PABX-BVA 418-013:PAGE 1-166 Page 165 Peak Area Report o [SamS s am:l xCot rts oeAi cq 12009800 rumensam PIA a " fom3ent Coaie lewthaol wfm taamni epee ried Worst Prtumber PAB BVA PrpS---- fa ogcooruge:IE muRmEpA NaEcossiI zntsn rst ieLbmeCoseeEstI T EEA SEaT-- as oma,a ereteTeseen re-t : ii { o i | BwnL[roearr | T I:\ T ! a TE TET 7. 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Page 167 Peak Area Report NoSuomRp eamm:2PxAR Ai5t52 oun AEcqrbionn8000 023 mensounpTeiiataamae:cASArAEeCAATAPABPABLOVAY O21, nusuVmmaanamameen:S507 onpedt Commpiam Impl i team heim Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-169 Page 168 Quintication Regore (QT Reviewed) @ RDiSaohcaofemil"e1:D38:0eDlra-DnaNkoT0Ana\cPAeBra\iPaARK BVA\L-062.110622056.S0EpaEciVcioarlt: w58a G45antencHegartahcaion BpaFr\asRsF:GRHTNE1L\TN.7ETHODS\BPVAABKW (RTE Integtator) [sl i = i A =Vv STNi ASEYAYA - wfone : 131 (0000) 4 rm? is i a Crm i -- er ---- I i AAT Nm IN RR 1 i| Te TE ThTieh vsih is rri | TEee ie eeik i TE Te TeTS Te Te Te1h tk ie oeat0se.0 PASIAN Wed oct 18 12:58:28 2000 rage 2 PrimediaWorse Project number: PABXBVA 418-018:PAGE 1-170 Page 169 Peak Avs Report @ LIES I menSRIS So Aman mnSRR 3 Wr BET Primedica-Worceser Project umber: PABX-BVA 418-018:PAGE 1-171 Page 170 Quancication Report (G7 Reviewed) @:ifDEaEtoaNFrilea1yD:\WeSCD_nDrAtTrA\aPweAaBt\ePrABE.BVAVE-062- 1\0621055.0SE_pearavtioarl:: 5span a#5 n1ntTesEeRaetiAoTn pazRanesc:h RNTEIEN.FP RGPADXBSVA\M (RTE Incegeacor) ses ms f | la: II i fhreb-- He neo rEo-- eET sEE-- EEEE-- TETTE neTRaTTR E Te Te T TT-- re - neni Lnee A| OLE neefnemo:6n%53ss69wroka eemsner m 4m im m 70 i| m 7% Te TE vw ih1 1 SeTTT | I -- -- tN | La inaeTa ako 4% am sw rm 7 7A 7% 7m 7h 7k7% TR 1m | 0GEI0N9.0 PABBA Wed Oct 16 12:50547 2000 sage 2 Primedica-WorcesterProject number: PABX-BVA 418-018:PAGE 1-172 Page 171 Poak Area Report NomsSatmplee:2A51D01Ovi552 owe pecotsy101020092 eo.SAA SAPABPABKEVAL O21 umenamo congue commit Imi) afm Se pea tne E800 1250 frat Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-173 Page 172 Quantitation Report (07 Reviewed) sBEaotaNneFile+1LDR:eTSeD DATRe\PrABN\EYABKBYR\1-063-1\0622060.0.S_fperiaVtioaar:, 0tow @ ir 45 tacEegeAscsTon TE aress: TSEWErS honssha va. (37% Integrator) F = r R \ r | | I - I\ | | MmL GwaasaEm$:55487 TR a we IAN a 4% 8 The TR Srount: | TR Te Te 0.00 7h TH 7% 7 --i :i Ai [R iE =at 1 aT e= E Ri RE m TEm TTeiea a hee A | JN | hiasn ek ee Eh ew em 1% 7% 7B 7% 76 TR TE TH 7%TH Gczion0.0 ThE SAN wed oct 18 12:59:08 2000 nage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-174 Page 173 Peak Area Report NorSouFmeiets:H20o5a02nvan2 uefS essn0200062 nseempadeieem,nCEASTtBAPAADEADLOVALOE meni 61 omgant Commpiins Imo) fn Salm imme -- sie 418-018:PAGE I-175 psn sermon mEoe |ANRETY Si E [ S Fram TERISanTv Enr or f. 7 Ne | ha! I | oAees RET = i i. J | rasp: 781 | A @i| snen ee em= em ah am ek fie io 1% 5) 1% 7HTooTm 167m im i Hi\ i Ih on imak ee e n ek sw ik ik vm3% ve TR TE I TR 1B | PrimedisWorcs Pret number: PAB BVA AISOIPAGE L176 Page 175 Peak AreaReport o STE SapteName: 2x STO-3Wate Co `DateAcquired: 10102000941 w TATr E Tonamecmncr sonREET com3ers com=pouns SZTEDLwp =sma spn Hr ons Primedica-WorcesterProject number: PABX-BVA 418-018:PAGE I-177 Page 176 Quantitation Report (07 Reviewed) PY DRaecgaaRnFileE+ D3:6\oKSeDa_uSAoToA\PAsDt\aPnARK-BVAVA-0G2-2\0621062.S0e peraViioai rlt: t2in 4a5 1atTegReEatAioTn 5sacanaS:intKrEMnE.R3GE AXBVA. (KTS Snegracor) for em | 1 - i | fp |refsopn:: 1133108 (7,13 Te TT TRA 200 A | " Fontees ie oe a oe ee TeTRsiTh 1-- ik Th re vee i resp: sam | i ji| || foo JN . | Ton ee eek eTTe ere dis rk i ik ia) o6aios2. mmevan wed oct 10 32159147 2000 nage 2 Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-178. Page 177 Poak Area Report NoboSumRpentueme::216BT04A1 O ecStas m085 1001 ee ATI UESones, ar 3 Som3at Cnwmigtelan lWoolon beWiwm aime Primedica-Worcester Project number: PABX-BVA 418-018:PAGE 1-179 Page 178 Guaneitacion Report (07 Reviews) @ DAfaetgatonPile :1 D3:6\NoGeDraDA0ToA0\PADd\0PtA00RK-BVA\L-062-1\0621063.0S_fpoarraaVtioarlt: t6)an G#5uaInntWeEstzhaotaion BrrorRaPnCs:HENRTIE\DTW.EETHSAOGXOBYSA.\M (RTE Integzator) rs we -- [= i | il jTi e0i1m5GeT aan vw 45 0h 6% 7ReiTeneTR 7% oToen 7 ve7h Te ik |reap 1517 , ] i ifh | } /\ | LLi tsk n se in wm an ew am Tm 1H%e1aht170sh1% re th rk vrkn | Resp: Sos . i Ifi ;| = WA ee hd i on on em xGman om ew 1% 1 ik tk re Th TE Th 1% 18 0621063.0 PASKEVAM Wed Oct 18 13:00:06 2000 sage 2 PrimateWorst Project mbar: PABX-BYA 418-018:PAGE 1-180 Page 179 PeanRepkort mimi Em oo EES ssn, on toe rs I. Primedica-Worcester Project number: PABX-BVA 418-018:PAGE I-181 Page 150 Quantitation Report (GT Reviewed) DB ataarFeE ileEDR :\WR SDR _DATA\PAAS\ABK-BUAVE-063-1\OR21064.0_B_gpaorsViinatl]: 6t4an @ ET L15 AsaTcegEra?cion S parass: TENRoEa os oarsv (kre tgeacon one1 meee TEi R --| -5 [1it i wP[w=oeInaspr: 31312wr0.130ra)ray ara ITe Ei NWe EN =WPT TRI = || ih i A | pi sk sk sk 64mm TE EER ie tho 7k 7h 1h [| na i---- -- || | Ii | e LahTeser e i aam am Te we Tr R reTe 7 7H 78 Tm ke Geaosen mR SAN Wed oot 18 13:00:36 2000 [I PrimediaWorse Project umber: PABXBVA 418-018:PAGE 1-182 rom J J SampleName: 2 x STO Wate: Date sr Acquired: 10182900 10.40 enEE eerasemen. wn ES nt capine SHE em cope Primedics Worcester Project number: PABX-BVA 418-018:PAGE 1-183 Page 182 Guaneitasion Report (GF Reviewed! oiDrHatalEPike + BY REEDIAETA\PTAS\GRSK-BVRVL-062-AOGRA065.0.D_r ertviitaE hls: 25, NET e aia--ioos sen ova 4 (478 Tneprutor) = ra om fi | El It i = rE I EEEA EERE E" AA we wR ! SR |Lo is oa [" a oo I | JA _ eh ri 1h Th Te ve rm vm 7m iE |i || 1 N FA) | Len ew en em sm em em ew tw 1 1h 1% Te rie 7k im 7h TE oai06s.0 PASIAN Wed out 18 11:00:43 2000 ee PrimateWorcstr Prt ub: PAB VA 418-018:PAGE1-184 rege 19 Pear aReker 0 mmm, eg ir InstrumentMethodName:PAEX_GVA NE... on tm T TE WEE Primedica-Worcester Projet number: PABX-BVA 418-018:PAGE 1185 Page 184 uansitacion Report (G7 Raviawea) Fonos @ iDAaetnaonFile+:1 D:3\NSD5DGKeT-A2\0P0A0D\P1A0R1K6.8BVA\L-062-1\0621066.DOEp_eTraVtioarl:. u85x 4G5ianItntMeegtrhaotdion 5sacAanCs:HARTEEWDETEHDODS\BABEBVA. (RTE Integrator) -- i | a,- |i iP = TIa E TTEE J TR rTrTTrTerry RoTmop:rr0i33 R| R -- h | i EAinR ikaeA Tw Tie T Th Th S 74 76 : TE TR TR rr | resp: T 62x se . il i (E TTEea E iin iEaE k Te R Te TE TTRET a Th ve Ta RTe Ta 0210660 PABK SVAN Wad Oct 36 13:03:04 2000 page 2 418-018:PAGE I-186 FINAL REPORT GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR ARGUS STUDY 418-018 Primedica Project Number: PABX-BBA PRIMEDICA ARGUS 905 Sheehy Drive, Building A Horsham, PA 19044 ~ 2 PRIMEDICA 418-018:PAGE 1-187 FINAL REPORT `GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA RAT PLASMA FOR ARGUS STUDY 418-018 Primedica Project Number: PABX-BBA Submited to: PRIMEDICA ARGUS 905 Sheehy Drive, Building A `Horsham, PA 19044 PrimeSduibemaitWteod bry:cs `Worcester, MA 01608 Primedica Report # PABX.BBA01-145 Page 1of 187 Issue Date: November 7, 2001 A Nes De LL @ DUBE 11/7 Project Scientist Bioanalytical Chemistry Department Primedica-Worcester PRIMEDICA PrFiinmaeldRiecpao-rWtorcester Project Number: PABX-BBA 418-018:PAGE 1-188 Primedica Argus Study 41P8-a0g1e82 COMPLIANCE STATEMENT interpretationofthe results in this report. `This project was conducted in compliance with the Good Laboratory Practice Regulations, 2ICFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision/ recommendation on compliance with principles of good laboratory practice. Official Journal of European Communities: Legislation 32 (No. L 315; 28 October): 1-17. Therewerenodeviations from the aforementioned standards, which affected the quality or integrity of the project or the DAAEIAL yo Mark A. Netsch, B.A. / Date Project Scientist PRIMFDICA Primedica-Worcester Project Number: PABX-BBA Final Report 413-018:PAGE 1-189 Page 3 Primedica Argus Study 415-015 QUALITY ASSURANCE STATEMENT "The following are the inspection dates and report dates of QAU audit/nspections for GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA PLASMA FOR ARGUS STUDY 418-018, Project Number PABX-BBA. Critical Phases 1. Laboratory Procedure 2. Raw Data 3. Draft Final Report Date Inspected 01/23/2000 03/29/2001 0419/2001 Date Report Submitted to Study Director ~~ Management 04272001 0112972001 0412772001 04/03/2001 042772001 042612001 The Final Report GAS CHROMATOGRAPHIC-MASS SPECTROMETRIC ANALYSIS OF MEVALONIC ACID IN EDTA PLASMA FOR ARGUS STUDY 418-018, report number PABX-BBA-01-145, was reviewed for compliance with the Good Laboratory Practice Regulations, 21CFR, Part 58, Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997 and Council decision recommendation on compliance with principles of good laboratory practice. Official Journalof European Communities: Legislation 32 (No. L 315; 28 October): 1-17, on 11/07/01. The results as presented accurately reflect the raw data. Noa Quality Assurance Auditor _uleals Date GPRIMEDICA Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-190 Page 4 Primedica ArgusStudy 413-018 TABLE OF CONTENTS PageNo. COMPLIANCE STATEMENT css 2 QUALITY ASSURANCE STATEMENT........ srnim--------" LIST OF TABLES AND APPENDICES... S CONTRIBUTING PERSONNEL ....crr simmers ANALYTICAL STANDARD CHARACTERIZATION/STABILITY ....cocorcs1 ARCHIVAL STORAGE wma SUMMARY... ------------ a) SAMI RECEP... sneer Calibration Standard ACCUanrdPTAECCISIYON ....ecvecrrrmsmrr orn Quality Control Sample ACCUTACY aNd PFECISION .....corcrrrnrnrrr rene 10 Study SampleConcentrations....... mmm----" CONCLUSION Summ J OPRIMEDICA Primedica-Worceser Final Report Project Number: PABX-BBA 418-018PAGE L191 Primedica Argus Study 418P.a0g1e8s LIST OF TABLES AND APPENDICES Page No. TABLE 1 Summary of Back-Calculated Mevalonic Acid Lactone Calibration SANGRIA CONCORTRONS corr smnssisn|2 TABLE 2 Summaryof Interpolated Mevalonic Acid Lactone QC Standard CONCERTAONS rvs3 TABLE 3 `CSOuNmSmIaArNyS oafndMeAvRaBlIoYSnIiSc DAAcEiSd.L.a.ctone Least-Squares Linear Regression | TABLE 4 Summary of Mevalonic Acid Pasa SEY SAMPLES. Lactone Concentrations in EDTA Rat 1S APPENDIX A Representative Chromatographic Data-Batch No. PABX-BBA-1-047-1.........23 PRIMEDICA Primedica-Worcester Project Number: PABX-BBA Final Report 418-018 PAGE 1-192 Page 6 Primedica Argus Study 418.018 CONTRIBUTING PERSONNEL Project Scientist - Mark A. Netsch, B.A. Director, Analytical CEISIY cco ..Jim Jersey, Ph.D. ANBIYHCE] CREMISL....rorse YER Villalobos, ACE. Report Coordinator mmm------------EEA L. Dicks PRIMEDICA Primedica-Worcester Project Number: PABX-BBA Final Report 418-018:PAGE 1-193 Page? 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: IntemalStandard: 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 4730071 50498 53 C, desiceate, store under nitrogen September 13, 2002 (Assigned by Primedica) September 13, 2000 5 grams `Aldrich Chemical Co. DL-Mevalonolacetone-4,4,5,5-d, MVL-d, Clear liquid U-295 215C September 11,2002 (Assigned by Primedica) September 11, 2000 0.1 gram `Cambridge Isotope Laboratories Characterization and Stability: The characterizationof the analytical standard and internal standard is the responsibility fabrication or derivation and stability ofthe Supplier, determination. as is the method of synthesis, PRIMEDICA FPirniamledRiecpao-rWtorcester Project Number: PABX-BBA 418-018:PAGE 1-194 Primedica Argus Study 41P8a.g0e188 ARCHIVAL STORAGE Archival Storage: The original Final Report and raw data will be `maintained fora minimum period of five years following submission of the final report in the Primedica-Worcester Archives in Worcester, MA. After five years, the Sponsor will be contacted for disposition iRnesptorructt#ionPs.ABXAr-cBhBiAva-l01ma-t1e4ri5a.ls will beindexedbyPrimedica-Worcester SPRIMEDICA PrFiinmaeldiRecpao-rtWorcester Project Number: PABX-BBA 418-018:PAGE 1-195 Primedica Argus Study 41p8-a0g1e8d SUMMARY: Sample Receipt: Samples in this project were received in good condition from Primedica Argus Laboratories in four shipments between the dates of December 5, 2000 and February 13, 2001. The samples were stored at <-20C until the timeofanalysis. 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 conversionof mevalonic acid (MVA) to mevalonic acid lactone (MVL) and the analysis ofa liquid/liquidextractby 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 intemal standard, MVL-d,, was used for quantification. `The method was validated for the extraction ofmevalonic acid lactone from 100 uL EDTA rat plasma samples over a concentration range of 10.0 ng/mL to 250 ng/mL. The complete description of the assay and summaries of its performance during the validation can be found in the report cited above. Sample analyses for this project were initiated on January 2, 2001 and completed on February 15, 2001. See Table 3 for the analysis dates of each batch. A copyofrepresentative raw chromatographic data from the first batchof sample analyses is provided in Appendix A. Calibration Standard Accuracy and Precision: Duplicate and bracketing seven-point calibration curves were prepared and analyzed with each batch of samples in an analytical run. For each analytical run, a 1/XTM-weighted least-squares linear regression was performed on the combined calibrant data set. Calculated correlation coefficients (r) for the regressions performed were 20.996 (sec `Table 3). Summariesof back-calculated calibration standard concentratiaornes 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 r`uSntsa,ndard Deviation (%RSD), ranged from 2.2% to 4.9% across concentrations and analytical PRIMEDICA 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: `Summariesof 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 limitofquantification, were labeled as "BQL" (below quantitation limit). Some samples reported as BQL were analyzed with a dilution but due to limited sample volume could not be reanalyzed. The detection limit for these samples is the lower limit of quantification (10.0 ng/mL) multiplied by the dilution factor. One samples where the interpolated concentration was greater than 250 ng/mL before adjustment for the dilution factor, the assay upper limit of quantification, was labeled as "AQLY (above quantitation limit). This sample could not be reanalyzed du to limited sample volume. The result for this sample was estimated to be greater than 12,500 ng/mL (250 ng/mL multiplied by the dilution factor of 50). Interpolated concentrations for all remaining samples are expressed in ng/mL. Refer to Table 4 for concentrationsofmevalonic acid lactone in rat plasma study samples. CONCLUSIONS: All analytical results were within acceptable limits with the exception ofone value reported as an AQL estimate due to insufficient sample available for reanalysis. Primedica-Worcester Project Number: PABX-BBA A18018PAGE 1197 Page) TABLES Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-198 Page 12 TABLE | SUMMARY OF BACK-CALCULATED MEVALONIC ACID LACTONE CALIBRATION STANDARD CONCENTRATIONS Batch ID 100 20Th0eoretica3l0N0ominal$Co0ncentrat%io0ns (ng/m1L)0 20 PABXBBALOSI 0N1R 210909 9N6R 448869 99469 1150 22849s PABXBBALOSSI 104 198 303 484 884 150 246 9% 193 301 slo 91s ss 2s PABXBBAL0SG1 916083 210908 330029 4s9i0z 889148 115500 224584 PABX-BBALOSSI 985 213 307 481 894 12 249 968 206 309 481 870 12 248 PABXBBA10621 973 202 305 486 82 1 242 NR 205 310 00 912 Isl 244 PABXBBA106S1 10014 119866 330059 44667 893586 11469 225555 PABXBBAI0671 99397 21994 33042 4m618 $9200 114s 223s5 PABXBBA-LOS-1 914 190 296 522 908 15s 239 07 206 324 46 86 147 240 PABXBBA-L0721 100 197 304 485 921 1a 239 NR NR ONR S13 920 159 246 PABXBBA107S1 9N9R7 1N0R1 334110 446515 898L60 11465 22591 MSde.aDnev. RSnD. o&Nominal 0939956 0290702 308 105 490 202 01 237 12 508 247 554 4106% 4198% 3148% 41%0 206% 303% 2220% 99.5% 100.0% 1027% 979% 102% 101.3% 98.6% NR = Calibration standard excluded from regression, data nt reported | Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-199 Page 13 TABLE2 SUMMARY OF INTERPOLATED MEVALONIC ACID LACTONE QC STANDARD CONCENTRATIONS Bach ID `Theoretical Nominal Concentrations (ng/mL) 20 0 0 PABX-BBA-1-047-1 21 73 199 sie 612 184 PABX-BBA-1-054-1 2s 654 183 29 00 187 PABX-BBA-1.056-1 2s 652 184 29 659 182 PABX-BBA-1-058.1 22 681 189 23 682 191 PABX-BBA-1-062-1 20 642 187 21 65.5 188 PABX-BBA-1.065-1 29 641 184 265 640 192 PABX-BBA-1.067-1 24 668 183 2856 684 182 PABX-BBA-1-069-1 2225 66614 187 188 PABX-BBA-1-072:1 22121 66918 200 192 PABX-BBA-1.075-1 203 616 18 239 6s 9 Mean Sd. Dev. n RSD Nominal 29 213 66.5 302 187 546 19 39% 20 45% 20 20% 95.6% 95.0% 93.6% *Gr=ubFbasi'leTdestto.meVeatluteheexacclcuedpetdanfcreocmritseriua amndmsdteaatteirrstmiyicnsed to be a satistical outlier using 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 PABXBBA-L047-1 PABXBBA-L-0S-1 PABX-BBA-LOSG-1 PABXBBA-LOSS1 PABX-BBA-LOGL1 PABXBBA-L0GS1 PABX-BBA-106T-1 PABX-BBA-106-1 PABXBBA-LOT2-1 PABXBBACLOTS-1 Slope 00067118 00062552 00061528 00060042 00061041 00064579 00064673 00063214 0.0064268 00069212 Intersept ~~ConelationCoeficent () AnalysisDae 0014046 099956 ovoz01 0.007087 099953 ovo 0.007974 099965 ongol 0.006094 099909 ozson 0.006886 099954 on2601 0.008650 099802 020101 0.002970 099698 o20m01 0.00018303 099789 20701 0.009049 099919 301 0.01975 099652 os01 Primedica: Worcester Project Number: PABX-BBA 418-018:PAGE 1201 Page 15 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES. Animal#Group Generation Concentration (ng/mL) BachiD 1100990023.-GGRROOUUPPII--FFOO 1100990045--GGRROOUUPP1I--FFOO 1100990067--GGRROOUUPPII--FFOO 1100990089--GGRROOUUPPII--FFOO 1100991101--GGRROOUUPPII--FFOo 1100991123..GGRROOUUPPLLLFFoO 10914-GROUPI-FO 21733 7 PPAABBXX..BBBBAA.-11..005544.-11 22630 PPAABBXX--BBBBAA--11--005544--11 230680 PPAABBXX-BBBBAA--11.-005544--11 33261 5 PPAABBXX-BBBBAA1-1.-005544--11 235017 PPAABBXXBBBBAA--11-005544.-11 314928 PPAABBXX-BBBBAA--1L-005544--11 253 PABX-BBA-1-054-1 1100990021.-GGRROOUUPPIL-FF11 1100990043-.GGRROOUUPPIL-FF1I 1019090056--GGRROOUUPPII--FFLI 1100990078..GGRROOUUPPLI--FFII 1100990190-.GGRROOUUPPIL-FFII 1100991113--GGRROOUUPPLL--FFI1 33698 BQL(DF = 1400,8<100 ng/ml) BOL (DF = 4104,3<100 ng/mL) 21476 2a252 311616 PPAABBXX-BBBBAA--1L-00S544--11 PPAABBXX--BBBBAA--1L-00S5A4--11 PPAABBXX-BBBBAA--11.-005544.-11 PPAABBXXBBBBAA-11.005544--11 PPAABBXX-BBBBAA--11--005544--11 PPAABBXXBBBBAA-1L005544..11 1100991165--GGRROOUUPP22--FFO0 1100991187.-GGRROOUUPP2-.FFO0 1100991290--GGRROOUUPP22--FFOO 1100992221.G.RGORUOP2U-PFF2OO 1100992234..GGRROOUUPP22--FFOO 1100992257..GGRROOUUPP22--FFOO 10928.GROUP2-FO 21350500 PPAABBXX.-BBBBAA--11.-005588--11 11510300 PPAABBXX-BBBBAA-110-05588--11 11815400 PPAABBXX--BBBBAA--11--005588--11 6927 PPAABBXX--BBBBAA--11--005548--11 x159 PPAABBXXB.BBAB-A1--10.5045--11 1271880 PPAABBXX..BBBBAA--11..005545.-11 2060 PABX-BBA-L0SS-1 BAOQLL== BAebloovweqquuaannttitaattoionnlliimmiitt DF = Dilution Factor Primedica- Worcester Project Number: PABX-BBA 418-018:PAGE 1202 Page 16 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal#Group #-Generation~~ Concetrai(on ng/mL) BachD 1100991165--GGRROOUUPP22-FF1I 1100991178..GGRROOUUPP22--FFII 1100991290-.GGRROOUUPP22--FFI1 1100992212-.GGRROOUUPP22--FFII 1100992245-.GGRROOUUPP22--FFII 1100992278-.GGRROOUUPP22--FFI1 AQL (est2i9m3a0te0d 23,200) 1157100000 2142900000 71618 13724600 18721100 PPAABBXX--BBBBAA--11--005588--11 PPAABBXX-BBBBAA--11..00662.211 PPAABBXX-BBBBAA--11--006528.-11 PPAABBXX.BBBBAA--11005548..11 PPAABBXXBBBBAA--1L00SSSS..11 PPAABBXXBBBBAA--11--005682-.11 1100992390--GGRROOUUPP33--FF0O 1165780 PPAABBXX-BBBBAA--11--005487.-11 1010993312..GGRROOUUPP33--FF0O 11244200 PPAABBXX-BBBBAA--11.-005588.-11 1100993334--GGRROOUUPP33--FFOO 1846460 PPAABBXX--BBBBAA--11--005588.-11 1100993357--GGRROOUUPP33--FFOO 1151300 PPAABBXX--BBBBAA--11-.005588.-11 . 1100993480.GGRROOUUPP33--FF0O a1s3t80 PPAABBXX--BBBBAA--11.-006528--11 1100994412-.GGRROOUUPP33--FFOO E4E16 PPAABBXX-BBBBAA--1L-00S5S8.-11 1100992390..GGRROOUUPP3--FF11 1100993321.-GGRROOUUPP33.-FF1II 1100993343-.GGRROOUUPP33--FF11 1100993357.-GGRROOUUPPS3.-FF1I 1100994402--GGRROOUUPP33--FFI1 25515060 PPAABBXX--BBBBAA--1L-00S487.-11 289570700 PPAABBXX--BBBBAA--11005S88.-11 199343000 PPAABBXX-BBBBAA--11.-005S88.-11 1133940000 PPAABBXX--BBBBAA--11005388--11 122700 PPAABBXX--BBBBAA--11--005588--11 BAOQLL == BAebloovweqquuaannttiittaattiioonn lliimmiitt. D+F= =EsDtiilmuattieodnvFaalcuteo,rDF = > 12.500 ng/mL. Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-203 Page 17 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES. Animal # Group #-Generation~~Concentration(ng/mL) BachID 10943.GROUP4-FO 10944.GROUP4-FO 10945.GROUP4-FO 1100994467-.GGRROOUUPP44--FFOO 10948.GROUP4-FO 1100994590..GGRROOUUPP44--FFOO 10951-GROUP4-FO 10952.GROUPA-FO 1100995534..GGRROOUUPP44--FFOO 1100995556--GGRROOUUPP44--FFOO 133 PABX-BBA-1-047-1 mn PABX-BBA-1-047-1 1590 PABX-BBA-1-055-1 1266000 PPAABBXX--BBBBAA--11--005588--11 161 PABX-BBA-1-047-1 177600 PPAABBXX--BBBBAA--11--005586--11 1560 PABX-BBA-1.058-1 99.1 PABX-BBA-1.047.1 1106000 PPAABBXX--BBBBAA--11..0085--11 22040300 PPAABBXX--BBBBAA--11.-004578.-11 10945.GROUP4-F1 10946-GROUP4-F1 1100994479.-GGRROOUUPP44--FF11 1100995501--GGRROOUUPP44--FF11 1100995534--GGRROOUUPP44--FF11 25300 PABX-BBA-1.058-1 2900 PABX-BBA-1.08-1 2015630000 PPAABBXX-BBBBAA--11.-005588--11 2013900000 PPAABBXX--BBBBAA--11--005588-.11 1189010000 PPAABBXX--BBBBAA--11--005558--11 1111003219--GGRROOUUPP1111--FF0O 11032-GROUP FO 1111003334-.GGRROOUUPPIII1--FFOO 11035-GROUP11-F0 11036.GROUP11.F0 11037-GROUPLI-FO 11038-GROUPH-FO 651947 PPAABBXX--BBBBAA--11--004477--11 20 PABX-BBA-1.047-1 29s7 PPAABBXX-BBBBAA--11..005487.-11 373 PABX-BBA-1-047-1 80 PABX-BBA-1-047-1 380 PABX-BBA-1-047-1 26 PABX-BBA-1-047-1 11029-GROUP11-FI 11031-GROUP11-FI 11033-GROUPL1-FI 11036-GROUP11-FI 11037-GROUPI1LFI 174 PABX-BBA-1-047-1 na PABX-BBA-1-047-1 202 PABX-BBA-1-047-1 17s PABX-BBA-1-047-1 157 PABX-BBA-1-047-1 BQL = Below quantitation limit AQL = Above quantitation limit DF = Dilution Factor Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-204 Page 18 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES. Animal #-Group #-Generation Concentration (ng/ml) Bach ID 11040-GROUPI2-F0 1111004412--GGRROOUUPPI1122--FF00 11044-GROUP12-F0 1111004456--GGRROOUUPP1122..FF00 11047-GROUP12-F0 1111004489--GGRROOUUPP1122-.FF00 11050-GROUP12.F0 1111005521--GGRROOUUPP1122.FFO0 155 22383 31s 115589 25 BQL(DF=11,4<0I0ngiml) 149 220129 PABX-BBA-1.056-1 PPAABBXX--BBBBAA--11..005566--11 PABX-BBA-1-056-1 PPAABBXX--BBBBAA--11.-005566--11 PABX-BBA-1.056-1 PAPABBXXB-BBAB-A-11--005566-.11 PABX-BBA-1-056-1 PPAABBXX-BBBBAA--11.-003566--11 11040-GROUP12FI 11044-GROUPI2-FI 1111004465.-GGRROOUUPPII22-FFII 1111004570..GGRROOUUPPII22--FFI1 1111005512-.GGRROOUUPPII22--FFII 525 PABX-BBA-1.056-1 Ses PABX-BBA-1.056-1 311560 PPAABBXX--BBBBAA--11..005566--11 83773 PPAABBXX--BBBBAA--11..005566--11 535160 PPAABBXX--BBBBAA--11-.005566--11 1111008534--GGRROOUUPPII33--FFO0 11055-GROUPI3.FO 111100876--GGRROOUUPPII33--FF00 11058-GROUPI3-F0 11059-GROUPI3-F0 1111006601--GGRROOUUPP1133-.FF00 11062-GROUP13.FO 11063-GROUP13.F0 11064-GROUP13.F0 11065-GROUP13.FO 11066-GROUPI3-FO 258243 BQL(DF=1,<I0ngiml) 228088 294 367 115902 238 29 180 69 13 PPAABBXX--BBBBAA--11..005662--11 PABX-BBA-1-056-1 PPAABBXX--BBBBAA--11-.00566--11 PABX-BBA-1-056-1 PABX-BBA-1-056-1 PPAABBXX--BBBBAA--11-.005566--11 PABX-BBA-1.056-1 PABX-BBA-1.056-1 PABX-BBA-1-056-1 PABX-BBA-1.056-1 PABX-BBA-1-056-1 11083-GROUPI3-F1 320 11054-GROUPI3-FI 199 11056.GROUPI3-FI a3 11058-GROUPI3-FI 640 BQL = Below quantiation limit AQL = Above quantitation limit DF = Dilution Factor PABX-BBA-1-056-1 PABX-BBA-1.056-1 PABX-BBA-1.056-1 PABX-BBA-1.056-1 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 # Group #Generation ~~Concentration(ng/mL) Bach ID 11059.GROUPI3-FI 1111006602-.GGRROOUUPPII33--FFII 1111006636.-GGRROOUUPPII33--FFII 11501-GROUPI-FO 1111550023--GGRROOUUPP11--FFOO 11504-GROUP1-FO 11505.GROUPI-FO 1111550076-GGRROOUUPP11-.FFOO 11508-GROUP1-FO 11509-GROUP1-FO 11510.GROUPI.FO 11511-GROUPI-FO 1111551132..GGRROOUUPP11FFOO 11514.GROUP1-FO 11515.GROUP2FO 11516.GROUP2-FO 1111551187.-GGRROOUUPP22-FFOO 11519.GROUP2FO 11520.GROUP2-FO 11521-GROUP2FO 11523-GROUP2-FO 1111552245..GGRROOUUPP22-.FFOO 11526.GROUP2-FO 11527.GROUP2FO 11528.GROUP2-FO 11529.GROUP3 FO 11530.GROUP3-FO 11532.GROUP3-FO 11533.GROUP3-FO BOL(DF=2,<20ngiml) 71815 194 S19 21 32%052 BQL(DF=2,<20ngml) 107 238 215758 190 189 280 272 208 24 m1631 928 160 135 1a35 190 21176 896 356 m 126 a 1180 821 PABX-BBA-1-036-1 PPAABBXX--BBBBAA--11--005566--11 PABX-BBA-1.056.1 PABX-BBA-1-056-1 PABX-BBA-1-065-1 PPAABBXX--BBBBAA--11--006655--11 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PPAABBXX--BBBBAA--11--006655--11 PABX-BBA-1-065-1 PABX-BBA-1.065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PPAABBXX--BBBBAA--11--006655--11 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PABX-BBA-1-065-1 PPAABBXX--BBBBAA--11.007625-.11 PABX-BBA-1.065.1 PABX-BBA-1.065.1 PABX-BBA-1.065-1 PABX-BBA-1.065-1 PABX-BBA-1.072.1 PABX-BBA-1.065.1 PPAABBXX--BBBBAA--11..007627.-11 PABX-BBA-1.072-1 PABX-BBA-1-072.1 BOL = Below quantation limit AQL = Above quaniiation limit DF = Dilution Factor 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 # Group Generation Concentration (ng/mi Bach ID 11534-GROUP3-FO 11535.GROUP3-FO 11536.GROUP3.FO 11537-GROUP3-FO 11538.GROUP3-FO 11539.GROUP3-FO 11540.GROUP3-FO 1111554412-.GGRROOUUPP33--FFOO 11543-GROUP3-FO 128 PABX-BBA-1-067-1 8210 PABX-BBA-1-072-1 375 PABX-BBA-1-072-1 141 PABX-BBA-1-067-1 an PABX-BBA-1.072-1 2530 PABX-BBA-1-072-1 13 PABX-BBA-1-067-1 336180 PPAABBXX--BBBBAA--11--007722--11 47 PABX-BBA-1-072-1 11544.GROUP4-FO 11545-GROUP4-FO 11546-GROUP4-FO 11547.GROUP4-FO 11549.GROUP4-FO 11550-GROUP4-FO 11551-GROUP4-FO 11552.GROUP4-FO 1111555534..GGRROOUUPP44--FFOO 11555.GROUP4-FO 11556.GROUP4-FO 11557-GROUPA-FO. 70 1550 9720 164 521 494 a8 457 330880900 ++ 648 320 sa PABX-BBA-1-072-1 PABX-BBA-1-072-1 PABX-BBA-1-075-1 PABX-BBA-1-067-1 PABX-BBA-1-067-1 PABX-BBA-1.072-1 PABX-BBA-1.072-1 PABX-BBA-1072-1 PPAABBXX--BBBBAA--11.-007752.-11 PABX-BBA-1-072-1 PABX-BBA-1-072:1 PABX-BBA-1-072-1 11630-GROUP11.F0 11632-GROUP11.F0 11633-GROUP11-F0 11634-GROUPL1-FO 11635-GROUP11-FO 11636-GROUP11-FO 11637-GROUP1 LF T1636-GROUP11-F0 11639-GROUP11-F0 81 PABX-BBA-1-067-1 254 PABX-BBA-1-067-1 258 PABX-BBA-1-067-1 296 PABX-BBA-1-067-1 808 PABX-BBA-1-067-1 03 PABX-BBA-1-067-1 387 PABX-BBA-1-067-1 358 PABX-BBA-1-067-1 383 PABX-BBA-1-067-1 BOL = Below quantitation limit. ADFQL= =DiAlubtoivoen Fqaucanttoirtation limit *+ = Value was confirmed Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-207 Page 21 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA RAT PLASMA STUDY SAMPLES Animal#Group Generation Concentration(ng/ml) BachID 1111664401--GGRROOUUPPII22--FFOO 1111664423..GGRROOUUPPI122-.FFO0 1111664445..GGRROOUUPP1122-.FF00 1111664467..GGRROOUUPP1I22-.FF0O 1111664439-.GGRROOUUPP1122--FF00 1111665501.-GGRROOUUPP1122--FF00 11653-GROUP12-Fo a3432 PPAABBXX--BBBBAA--11.-006677--11 a1465 PPAABBXXBBBBAA--11.-006677.-11 2i5n8s PPAABBXX--BBBBAA--11--006677--11 23555 PPAABBXX-BBBBAA--11.-006677--11 119072 PPAABBXX--BBBBAA--11--006677--11 72436 PPAABBXX--BBBBAA--11--007627--11 381 PABXBBA-1.067-1 1111665545--GGRROOUUPPI133--FFo0 1111665567--GGRROOUUPP1I33..FF0o 1111665589--GGRROOUUPPI133..FFoo 111166610-GGRROOUUPPI133-FFOo 1111666632--GGRROOUUPP1133..FF00 1111666645--GGRROOUUPP1133..FF00 11667-GROUP13.F0 32045 3105s BOL(DF=11,8<10ngiml) 83112 21608 20 BBQQLL((DDFF<=11,,<<II00nnggmmll)) PPAABBXX--BBBBAA--11--006699--11 PPAABBXX--BBBBAA--11--006699--11 PPAABBXXB-BBAB-A-11.-006699--11 PABXBBA-1.069-1 PPAABBXX--BBBBAA--11..006699--11 PPAABBXX-BBBBAA--11.-006699--11 PPAABBXX--BBBBAA--11.-006699--11 11510115/10145-0G6R-OGURPOLUPFL1LFL 11510125/10155G0R7O-UGPRLO-UPF11-FI BQL(DF=41,5<440ng/ml) BQL(DF=31,9<230ngiml) PPAABBXXB.BBAB-A1-1.-006699.-11 PPAABBXX--BBBBAA--11--006699--11 1111550089//1111550130--GGRROOUUPP11LLFFII 11IISSII231111551114--GGRROOUUPPIILLFFII 22548 PPAABBXX--BBBBAA--11--006699--11 23 PABX-BBA-1.069-1 205 PABX-BBA-1.069-1 1111551175//1111552204.-GGRROOUUPP22-FFII 339830 BAQLL -= ABebloovweqquuaannttiittataitoinonlilmiimitt DF = Dilution Factor PPAABBXX--BBBBAA--11.-007722--11 Primedica- Worcester Project Number: PABX-BBA 418-018:PAGE 1-208 Page 22 TABLE 4 SUMMARY OF MEVALONIC ACID LACTONE CONCENTRATIONS IN EDTA. RAT PLASMA STUDY SAMPLES Animal#Group#-Generaion Concentration(ng/mL) BacDh 1111551291//1111551186.-GGRROOUUPP22-FFII 1111552253//1111552287--GGRROOUUPP22-FFII 11526-GROUP2-FI 227359 PPAABBXX--BBBBAA--11..007625--11 211542 PPAABBXX--BBBBAA--11.-006699--11 490 PABX-BBA-1.069-1 11115S3368//111513573-4G-ROGURPO3U-FPF3I1 11514105/4121-5G3R0-OGURPO3U-PFSI-F1 11543/11529-GROUP3-F1 230925 PPAABBXX--BBBBAA--11--006752--11 21093 PPAABBXX..BBBBAA-.11..006655--11 340 PABXBBA-1.072-1 1111555427-.GGRROOUUPP44--FFII 11116633021111663336--GGRROOUUPPLH1--FFII E3S1/1N6I386-3G7R-OGURPOLU1-PFI1LFI 11639/11634-GROUPLLFI 5313 22470 BBOQLL((DDFF-=22,,<<2200nngg/mmll)) 214 PPAABBXX.BBBBAA--11-.006699--11 PPAABBXXBBBBAA--11..006699--11 PAPABBXX-BBBBAA--11.-006699--11 PABXBBA-1-069-1 1111664430//1111664471.-GGRROOUUPPI1Z2.-FFII 1111664468//1111664541--GGRROOUUPPII22-FFII H6S1H1161506-4G2R.OGURPOLU2-PFIIZFI BQL(DF=42,3<d0ngiml) 211497 BQL(DF~22,4<20ngiml) PPAABBXX--BBBBAA--11.-006792--11 PPAABBXXBBBBAA11-006699--11 PPAABBXXB-BBAB-A-1L.-007629--11 6H64S16/116166525-GGRROOUUPPIISS-FFII BBQQLL((DDFF==22,,<<2200nnggiimmll)) PPAABBXX--BBBBAA--11..006699--11 BAOQLL == BAebloovweqquuaantniitaattioonn lliimmiitt DF = Dilution Factor Primedica-WorceteProject Number:PABX-BBA Pages 418-018:PAGE 1-209 APPENDIX A REPRESENTATIVE CHROMATOGRAPHIC DATA - BATCH NO. PABX-1-047-1 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1210 Page 24 Peak Area Report NotbaSnakpPoiaeene:s PuAsBBA 1087 oneeSqon 121 152 SAT SrA apper meen Gompadt Comgpue Img m tmami tapes ATor oe stmr no rtPt. Primedica-Worcester Project Number: PABX-BBA 4ISOISPAGE 1211 Page 25 0 STtI rL en , Ts, @ Method : D:\NPCHEN\1 \NETHODS\PABX_BIA.N (RTE Integrator) Wr mr oF REEEEEheRE nd A pag EE EEE TEEEEE Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1212 Page 26 Peak Area Report NobSoamRpoaanmen:1AxSH50A+167 owedocqutme 1201 154 enspaeds cowissmpm Imwao) sBm muem smo retour 1801 1480 vase 1 PrimatesWorst Project Number: PABXBBA Bia fet QL ET ene 418-018:PAGE 1-213 Fagen HI ET ET woo = Wael aie eB eh ek ie ie on EOS Rk Ikte 3% 3 ih te th ey R TREE R Nh RE Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-214 Page 28 Peak Area Report NesoSmuRneeaarme1PxA5B70B2A 41 oeArcaqnd 1201 1628 emeroobvaeaFli apumne oOArM1B0S2AA30TAPAGEASK BALSA. taenrVenettarme 57 ompedr Gonppilem Imfel BR hemi haa Ttmi he eean in spend et rg Primedica-Worcester Project Number: PABX-BBA fSonatriete : 0o1mD pATRenePABCAER -G47GATIO05puvriia 3 QT EAT naan a sneer 418-018:PAGE 1-215 Page 20 on TE TETRI_ RTRETeeTR TR to ow t oA o wT l5 h--eCMTR E TR Te TaTR eTe e Ths e TnI ounom.o maEMAK won 3 lesen 200 ras Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-216 Page 30 PeakArea Report NoseSsanFpeertnes::P1xAG5E70 1067 oweAeoqurme: 1201 620 versAESAT ADCBA 10 van A Comat Gonmptuns IMR fm eam Psusas FrawT 1801 485 [ rimedica- WorcesterPojct Nur: PABYBBA L1 r A ST ragYiu + TT 418-018:PAGE 1-217 Paget i AA -- ------_ -- Er T ve Se et hh TeTeh TgE veITe 7I h TehhR TTR aRa veWh ve aw 7m Tie vk ih ik 7h ve im ve ve | Primedica-Worcester Project Number: PABX-BBA 418.018:PAGE 1-218 Page 32 Peak Area Report Nee Saameea.me A1570C4. S67 oveciaer:20 047 en sTmieas:CABSRBADATAPKESABLSBAO07.1\ meen oS fomeapeds cwomaplum lewfo) wBm meAaRK Pome Pun wtTew 11801 1448 [ Primedia:Worcester Project Number: PABX-BBA 418-018:PAGE 1-219 Page33 STBrinAL ELREERSE1i00. QE Rrse oe ren F A e _ roo wlTlERRWh We AR aRjReTle Th Te 7h ik 7hevRe te hiee te EE EERE Primedica-Worcester Project Number: PABX-BBA 413-018:PAGE 1-220 Page 34 Peak Area Report NotbSounkmPoearnees::P1xA5K70S5 1087 oveAac:u20n1 1a707 en SETAE SU Snorapiesanarn min Gmmest 2 Comspoun wis Dain wm mesma caus Ea `Primedica-PWroojreccteNsumtbeerr: PABX-BBA Bata File 0:\W80 DRTR\PAG\PAGK-UAVE-047-1oK7096.9 vial: RERLARNT TY fa QT SHR ros east_ssa ne saves) 418-018:PAGE 1221 Page 35 FE TS TEEAEMPPP ae ph EgES ih a-- akan ak -- 3m7=7h7h--7--h 7--6--Th--Te----Th -- aoe 1 CORR NN aha a 7h 7k 7 7% ve 7a 7h TRThTh Primedica-WorcesterProject Number: PABX-BBA 418-018:PAGE 1-222 Page 36 Peak Area Report Naeooosakrapienasne: xA0O8A 47 oeAScnoe 12011727 rmaCoemotesa: ME5R,DAATAPABPABKSBALGAT, mesurevn] fomsjeds comple Impl Rum mpm J -------- SRS TA rest 418-018:PAGE 1-223 @ Method : :\NPCKEM\1\METHOOS\PABX_BAA.M (RTS Integrator) pp EEeReEeEeeEe TR i re Nn Te TTTeTame Primedica-Worcester Project Number: PABX-BBA 413-018:PAGE 1-224 Page 38 Peak Area Report NoSsuenetmtaacnse.:1F3A5S7S0a7 104 oveApcrqouta 12011747 Te SUESurname mis Gomp2edt comopnas InwgeombesinmE mie Primedica-WorcesterProject Number: PABX-BBA fmauisEerie RDoimgeYN smE n S7s TIONS5n,e48 + QT BR Eommasn 1 scr 418-018:PAGE 1-225 Page 9 Fe. NE gp -- ee w ETt E ih eRT lThep E a R TThR he o 220 i AN Tvi e eve TRTeTe a ve Th TRTe Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-226 Page d0 Peak Area Report NeSkaomr p amn e:P1 Axs cKoAnTR:OL47 Over(erd 201 1057 recConamdaoirrACBAES9TAACABEAISMTK nasavhs meer 2 hn compe wis DEED mo rem tems Sam we lc to otre pror or toso nTe 11601 1647 rages Primedica-WorcstrProject Number: PABXBBA 418-018:PAGE 1-227 Page 41 A B J zeE + 5 EEY5=0Ya. t 3 QERE EREEosmansun (8 seseaser tra HR RRR TE R a h RR r TR T eT e Primedica-WorcesterProjectNumber: PABX-BBA 418-018:PAGE 1-228 Page a2 Peak AreaReport NotbSaaokmFepelrees:: A1GxH08O1W047 owe Apcqad1201 1826 enemyBATA BESBTAPASPABLOG1O1A.mar ompgaedt Cowmipsan Imwplow eaamm baipges Aer ch in mrt gonoterr rg PrimediaWorst Proj mbar, PAB BBA BBy ENEg @ ET fret fEi e Sp -- 418-018:PAGE 1-229 [oY Toe6X826X se6% SE 8% ew 6 Tw 10 TE 7% Te 7h 81% 1% 1h Mg pga A eE d Eg Ta he EEE IE TEEEE TETEEErr \ i CEE| ET. I. Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-230 Page 44 Peak Area Report Nenesanpen: oogroeos0 aEd-- pe3 r Copoises Iswfoe wB MaiOwne Sui Tt on hnds tgtn rp Princes WorcesterProjst Number PABX-BBA 418-018:PAGE1-231 Page is B ge itRe 0E 080 m E aeeve o, a QTE Intn rs seen i" A A TR Mla em MS TR A @e e R Te S J eehe | N TIE ee ie Re ThTeeTe Teee TR RR primed Worcs restNahr: PABYEBA er 418-018:PAGE 1-232 PessoaBape PE [EO NaTm onIeE cn men Era ree no re pe ape tet maticsWorcs Project Number: PABX-BBA Pages? 418-018:PAGE 1-233 Be ARE EL QTE IOI mn one 2 een J ee TR TRE TR Fg meme et rem EE ECTET Er EE ne TRE vk we an ek ae rh 7% vie ve ve 7 ve 7% Te Th Primedia Worcs rjcNumber: PABXBBA 418-018:PAGE 1-234 Pages [-- 0 SspE p E nmges ps aii mo1ar sozme BE R ynE nage ppp PriWormstaProtj Ners: PABXB54 BSAi TIALRS rE eir RETR QTE ET Emons, eee 418-018:PAGE 1-235 Pages C a N EE mes 6 GR $% ee Sho SwWF ek 4% 150 70 1 1%14 16Te 7% te TE RRen Rh TRAh e A e | rime WorsatoePjNobo: PABBA 418-018:PAGE 1-236 _-- Pear pent 0 I[EREEE csfod wen RESI, ran, wnBRE me cme BE sme see re ee terre rere, Primica Worcester roe Number: PABXBBA 418-018:PAGE 1-237 Faget RE BE wE. QB I svwergo ra semis } Er hy or A Te TR 8 Tim we ee ie Oh ie we ve TTek TR TR 7a 7h Th 7h ih im TTR eR eekaean em ew 7h 7% 7m 1% 1 7% TE ih ie me Primedica-Worcester Project Number: PABX-BBA 418.018:PAGE 1238 Page 52 Peak Area Report [Sul mp at me t 34GHOU5R0 [ pred-- enAAT PLESrna 0, mi fonads Gomwgopintes Imwao) mdm hmuwm Maes Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-239 Pages saa pie. SAee iat 30 BER ear i QT BE unemarson 2 sesso =| La 8TR0 8 Pg i vn SR TRC EeTR 7 TR TR TeTR ARe Ra TR Wea wh ee iTTRTR Re T TRTR RTe Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-240 Page 54 Peak Area Report NebSoamRiaramees:1P0B53K5GSO0A037EO oveA= cq 12012026 senpoidetaPro:,SABSS ,G8S4ATAPABATK BAY411 arvams30 fampesr compas mie Rommime sae Twmn chrec atro opted emg Primedica-WorPcreojiecrt Number: PABX-BBA 418-018:PAGE 1-241 Page 55 r ESaEUT AT REA E TASwru2th .7 QT EST = renso prssomens errr N EE N eT I e JAFFEr Ce EHIEN BmEeNeNeety 8e3 do sm ow 6 6% 600 6% 70 110 7 10 6 t 1m tmiw| rimatica-WorceeProject Nahr: PABYBBA 418-018:PAGE 1-242 Pages Peakrea Report @ eS eSmNIS mEN otfia ms ce comTes co=mers ZEwl mame ce `Primedica-WorcesterProject Number: PABX-BBA Sg Sp BEE fit, QL ER nnnsn or rn a1s0ispAGE 1253 Page 57 ET a a fp RAE 6Jn Ra R a ae Nn TE TT A Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-244 Page 8 Peak Area Report NelsSeunmRpomaemr::P1A3O40BGERAO5U4P7E Acouprea: 201 2100 meteFdehaar.e SAS.SBATAPASSPBAA0B7K, Wer mem Gear AT ommplum Ima mR mlm ncneo a gtrsper mp Primedica-Worcester Project Number: PABX-BBA 418-018PAGE 1-245 Page 59 Esagtaa tile 5,000BEAmTA PARK BERTR030Sperkviiand] 32 QE DR | aimoosmax ow 7 Enceacor) P EET Te ve eeTe TR TR TRTE TR TR RR eS Ny REE i i eR ee ghTe TThk 7h 7h Tk ieRR A] @ = | A CRT RR ae ek ek ew ae ve th 7% ve 7h Th 7h TRTR ens mecHAN The dan 16 30.56.00 000 fase s Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-246 Page 60 Peak Area Report NotbSanakmRpeeam: 1F5A2X0S600009520751 onepocors 1212124. me coon EE mgm oye Primedica-Worcester Project Number: PABX-BBA sarin asses ers oy Tr QT Lhonsensn. (57 osegessor A15.018:PAGE 1247 Page61 i | ETT R ThTeTT oN TE ee Te TT TT TTR RR Te TeTR TR Primedica Worcs Project Number: PABX-BBA AIS0ISPAGE 1248 Page rmaPnear Report omg o NotebookReference: PABX-BBA-1-047 Operator: renCEIET SmRacspsn eS mest omg HD mame ean Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1249 Page 63 Ee DatLaEFile:e Dh:aD DBnARsTAR\PBARARVA\04B7-A1\RONKTIO.N F2SpEecVkiinahr: 34 er [opiate i L$" TREO VR vieTT7a7 7hhh P EEe kW ik WsWB g R Te TETe7 rh Te ih TTh h TE OR we vee eeamTe TeTh 1h ve 1 ve 7h mw wm Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-250 Page 64. Peak Area Report o NabSouoneoleamseA 1033B2o6B 80003851 OverSese: 12012209 eEn AT ESBrno ss, weVva e S| Gomspest Tu Somptens TR Meme Spans ho leStak to opr emp Primeica Worcs ProjectNumber: PABXBBA 418-018:PAGE 1-251 Pages L BE BIY f=et. QL ST en sn an R a R TR EE 0 PiEEREEER ETE Cm en akse em am em ew 7 77 2m is vk te 3k Tw am Primedica-WorcesterProject Number: PABX-BBA 418-018:PAGE 1-252 Page 66 Peak Area Report NottSeamFpeleern:A1xO05AuD1067 oneapcaets 201220 mensBTR SASSAPDPABAOAL 01 iG omnais Gowmisme Ifwao) wBn mueem Mages Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-253 Page 67 E DReultoanTMFilR e: D:\A WGEaDnD2A0T8A1\GAHBE\PYAOKBRON.1NOUTIONEDBTeoeitals: 13 QETRA Bu Wemcoe\s saa wean (x78 sesesrator) FE i i MEME NE NEMEME TE Bropr gresserece beS smsesris tre sevens, BJ R I m Ea ME EMA TR CTT Raae eham ak am7hk 7k 7hve 1 TR TR Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-254 Page 68 Peak Area Report [ORSumwo ihatme 10335 8009951 oeppceaeure 01 2.00 erCTE ESATA ADELA, warearon3 omipest | csmpuem Imfel BR OmUDT SMmpe T on ie aab grorntd meorts oeom ATX8A - 418-018:PAGE 1-255 ERNE @EE Jit SR TE ---- eee060 R 4X Gk 6% Se 6% eM em TT_ RTw TeT 1h 1k 7% Te TR Tih wh swaw sk ak_sk_sh Th TW 3k 7% ve Th TE Th Th 1% | epfA|| I 82 6X sw 6% em eM6 _4w 7m 7013 1% 14 1% vm 1R 7 m we Primedica-Worcester Project Number: PABX-BBA. 418-018:PAGE 1-256 Page 70 Peak Area Report NooSoampRleewannees:A68B3C0GSROOnUP4D11 J ovepores 1201 2202 ommaest Gwoimmepium Imadommo tama tame Primsica WorstPretNombr: PABX-B3A ET REE Er PE -- 418-018:PAGE 1-257 Paget . A ek a -- I E i EEEEEE h EEETEe aETT == TE ET AT a Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-258 Page 72 Peak Area Report NeSoumcp oamne:a0336GSRenOUoRtD1 ove epntn 121 022 mntTaeisrapaeCABSC5OAATAPADPABLIGAT O71) abuse vermemoe 8 2 wo wom tm Primed WoreesrProj Number: PAB 354 Pers 418-018:PAGE 1-259 OH Enns si nee810 63063 6 6%Sm EM 6m eT 101% 1% 7% 76 ve 1% 18 1m EE E ri N a oHH |i EE RT ee a ih ek th 3h Te ih Te ve Taih Th The rimetca WorcesterPrjst Nmber: PABXBBA 418-018:PAGE 1-260 fo Pook eaepon 0 EE ae sE om ongmreain OoOE 7 Eo HE ere sme rete rt et Primed WorsssProjNumber: PABX.584 TT 50 QoHRT ERS bamosaox s sw (85 tncracon) 418-018:PAGE 1-261 gers elrr TRRR ro| S23 sk sh a GT eR ew 01% 3B IW Te 1% te vm th 7% Primedia:Worcester Projct Number: PABXBBA -- 418-018:PAGE 1-262 PecArn port 0 Sn EIiT: E J Se R sA aE mI s SI sgnwen rTeRArTEeS 7 = 1% Prime Worcs Project Number. PABX334 rage 418-018:PAGE 1-263 BBO ENEEER 3bi0ns ET BR oma sx xe crac) : I RT a rd I TE Primedica-Worcester Project Number: PABX-BBA 413-018:PAGE 1-264 Page78 PeakArea Report NotaSsaRmapnlee::F01A2C3S080502750 oweAecquris 1531021 eeampaon ipane.SAuBsC8SXATAPABPABK BALO41, nme Tn Gom2eds Comwpwass ImTeo) wbn hmahema tte oe bhsh cd ota oonopr otrn. Primedia:Worcester Projct Number: PABX-BBA iEI eER50Espn-- oon t @- Wathod: D:\HPCHEM\1\METHODS\PASX_BSA.N (RTE Integrator) 418-018:PAGE 1-265 Paseo ETo RT h TTTEeR Wee Tahawwk ek am en eEml i sw Ta Tn 7h Ee 7% a3s ve 3% 7m 1ah d dd EE MMEMT| PrimediaWorcs Project Nbr PABX-BBA AIBOISPAGE 1266 pgs 0 Peak AreaReport PR soigipyn wenn EErwemasuonn, won o PrimiWorestrProject Number: PABX.BBA E Hi m| SHaATI TL OT SIT erasx me 418-018:PAGE 1-267 Pages er eee OS rEr rrarra itt r lf SR RR . | TEER eeN Re e Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-268 Page 82 Peak Are Report o NebSoamapnleers: 1A03B42S6A805015p0 ove ASocie: 1501 100 net eaTdoHaar.n:3ADSEh5,6C4ATAPABSPBAABK G01. sarVaR gomaett on Cowmimsies cnree IwmlowBoRdwmmE time gn rnd mg. Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-269 Page 3 SEatErite Du naohn as BERT\GE002fiint 3 E TSTp RE-- monnsoa x7 ncesrcen) ta I ER TRTETE RR Te a TT R Th h Niho TehTh Th eTh oT ------ Ch EE rarh rE h aaa] coronene en isn 200 oz PrimedicsWorcs Project Number PABX.BBA 418-018:PAGE 1-270 Pages Peakaren Report . PR. -- reBeErcmon reSTE 7 z= Ee PrimeticaVareseProj Number: PABX-BBA Lm g SEe gpg ----r= ot 418-018:PAGE 1-271 rages eeRE SEE R EE hao e da R TTTEJ EERRe TEa R REEh AER I| | PrimediaWorse Proj Number: PAB.BBA paess 418-018:PAGE 1-272 JSI o `Notebook Reference:PABX.08A-1047 a Operator. oo SEE nan wRon ES comTes ommeer EOZ EsmEe se er erp t ette Primedica-Worcester Projet Number: PABX-BBA sso 0 nanam o Toe ros gi 18 HE hm Be QT ESTE arom,sun re omens 418-018:PAGE 1-273 Pages? " NT ET TI TE RT VE ah E wh OR ih gah ile 7h 7h Te ve a Th Th A TERR a eeeh ue Th Tie TR Te ve 7h Th 7h TT h h PrimeicaWrest rjcNumber: PAB BBA 41501PAGE 1274 Pages PeakAes Report 0 IsaN m sE ome oes ssorm SEE InstrumentMthodName:PABX_BBA worn, wd " s7e copmixpe SE ETmaEgma me wrews-------------------- et nn es Primedica-Worcester Project Number: PABX-BBA 413.018:PAGE 1275 Pagess pa rite 5,\gp ETRE AEoenGETION 8 vil: 46 BLN dsm TE er ITE ETE Lon stn re sees ee EERE EEREA ER ET AEE TEE EF ee ~~ N petteOg TR eenme Re i ve TTTeTe Te Te TR Te PrimediaWorse Pret Number. PABXCBBA Fogo 418-018:PAGE 1-276 Pe rea Pups 0 SEs NTMm EEL sBsn TSS InsetthrosNuamme PeABnX_5tA sr wrERE cos cmmpiee EE wp nme ne Primedica-WorcstesPojct Numbers PABXBBA 418-018:PAGE 1-277 Pager SBL RE B=ut , rSTe ST--wangon 5 gens in ER ge I ee al Sl Pe R TER el ak sk sm an ewew vw 700 TmIm 7a 1% re ih im 7m rimedisWorcester Projet Number: PABX.BA 4I8018PAGE 1278 Fagen PeenRekpo o S-- um-- m rnye men ESessa. min / Ir---------------- ri Were m Projecte Number PAB BBA LBeL" |AREENei------_-- tt LE BE in sve a se ges 418-018:PAGE 1-279 i. I ETREAT EEE TEE Weer et n r p R ENRE NR EE SA o EE E h T Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-280 Page 94 Peak Area Report NottSe uFloaens: AsBBROA 04TD oned= ens: 1201258 J ey Gompageat Comwmipitans Teweol mfm miEmmi cei PrimedicaWorcs Project Number PABX-BBA Pages 418-018:PAGE 1-281 FEe EoRu mEen twte LTT ESmT a snr ae rego tn ] EA E TT TR ,| @CT ETEmoETeRTE R EEE R T R Ee E E ee EE A R he Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-282 Page 96 Peak Area Report o NolSamnRtoaeme:0F0O7O00P02650 owe oqidn: 1301310 meneoomdiaameSAoEiXSGAETAAPABEABXOSALGN, Warmer momen onsp2adc Cowmimsies laE falEBn otegm ulm rime Wore ProjectNumber: PABCBEA 418-018:PAGE 1-283 Fagen QWLhRieE EpE oTmR e ons TE er ee er be A A Boa tte EdE ER e WE welET EE - | Nr aa Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-284 Page 98 Peak Area Report NoSbunaolmamae: 1P0S35EG8A0UoP1b50 weAecarrd: 1201337 strat eamoid apro:.SASMK.,BSAATAPABPASKSRAL O71. ane memo 5 Compadr comptes Imi mode Ree J -- Primdica Worcester Pret Number: PABXBBA 418-018:PAGE1-285 Pages BEE TE TLET Oe TI aangon we apes i TR TT TET A We4 ik v8 gTkh uh 7h ve 7h 7h 7h Th Th Re Te TeRR Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-286 Page 100 Peak Area Report Semenmocoirro onecen: sam 357 J Gsndr | spies pr ef lm tint eRipe J ---------- Primedica-Worcester Project Number: PABX-BBA E gre N1 sEm yDey STte QE EEocrT n so 8 sgn 418-018:PAGE 1-287 Page 101 i EERE ERE EE EEE Wg ga I Se JC ol r AE er ET TET rimeWorcs Post Number: PABXBBA oeak apt @ IA EEE 418-018:PAGE 1-288 Page 102 ctte re 1 fo EE Primedica-Worcesier rojct Number: PABX-BEA pce ie 3 SAFnAesRacIin rOssEEroVsiaels 33 Bia Tet QE BE comma sun he pcs 418-018:PAGE 1-289 Page 103 Lee 4% AB_4%G6 SW Gm Gh ew ew TeTnTm TN7a 7% TR TR TR TR mr ee TT Ta TR a Rh TEE ee aTeTe Ta ee Te Te mh TE Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-290 Page 104 Peak Area Report NetStamopFle oam:rAe1G5H3SsERAOUvPo4b50 oe Aecers 1901436 ES SU Shes mar gesm2ess Gowmmsplem Imoplw me mpe Tr cottrgn rd gn Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE1-291 Page 105 Be imma = ppt A -- i A a T R ET ve ShiE b3sThiT 1% 7hTe 7h 1h Th th TR h EEha Te TR rimetica-PrWojoecrtNcabseer: PABX-B3A 418-018:PAGE 1-292 ---- PenkAreaReport 0 EPE--E omamno ion ar3 r cfoom)pe SrDin2ape pe ere Yr eAsrd PimedisWorcs rjc Numbers PABX.BBA E pe iE e 0R2s E p fta QTE Eensa 8 csr 418-018:PAGE 1-293 Page 107 eB a = ae te dD 1080. Th Te a 1 T NR NTehe TeRe rimeticaWorcesterPrefect Number: PABKBBA 418-018:PAGE 1-294 Fae 108 Poon 0 EERE renmeCSE M gon ggg enRS oer comp Em am eae FS, Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-295 Page 109 aaa rile 00DBYR AEEBAST NeNTISS 2 Vi: 55 HEE end Tr QsT erenaa . wr conone ww (18 epeaon) me a WE AR 44 SHGW 6% 6m 4% 1m 1%TR 1%1% 1% 1 1Th% TW i Ee iE ah wh vk wk en ek gmTh 7s TR 3% Th 1 3% 7h Th Tm ! \ || Tr E i r eR Th RTr Tee TT TR Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-296 Page 110 Peak Area Report o NotsbooSkaRmpelwee:r PAB88G8RAO1U64P7E AcOpacqror. 01535. os TI UE BAAPABPADCOB1A1I,wo a e4 Gommaadt AToe Comwminits ImTpel omo mTiume Sms tn et a tn ei rdor tr Primedica-Worcestr Projet Number PABX-BBA BEIER T fWhut LT ET wasn gin is goes 418-018:PAGE 1-297 Page 111 i oT h TT TR FE . FA ARR RAREEEEAE a EE TEE ee Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-298 Page 112 Peak Area Report NomsoSokaempelne:.P60A6B7B6A80o04s51 oweaScnoe io 5 - EE EASAaA sa marr5 mets cmmpum Imi dm tame mime . Primedia:Worcester Project Number: PABXBBA BaEaEepie osose FURR SpA AyLTER S( E--Ein-- 1er QrsT prasion EureTwanEecono tr seen 418-018:PAGE 1-299 Page 13 wf | Lome 30 SR 63 44 4% am Am em GK 7m1% 130 1H 1% 18 78 1m 180 tw Fh gy me eee TE eT TeT EE TER I TE eh ie TETeeT ee hh Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-300 Page 114 Peak Area Report np: ros oungoin 1301814 in ahFdiNe Fao SAt BORSSATAPABPAXBOALGH, WotmR Gomanar ceEmmapam Imwaod w ECuRe mide Ahb re mr prot dn gn FetomewsTme 11851 500 [ rimdinWorssis Profs Number: PAB.324 fBBEgESEIERRBEpo ---- t $BSisL QLAE-- R av or cr pope 11s 418-018:PAGE 1-301 Se = Em Ben lf BT RR RR C EE ETEEN EE eh rimedis-Worcester Project Number: PABX.BBA 418-018:PAGE 1-302 Page 16 [---- @ o spnI oy omi=g=mimien nrSTEMI yess nS err seme SHED gy cme egg vam vormo---------------- ntsNor rsae PABKB98 co ss enTL ron ah LR Toe ws ET BAWe aho\o PAS s 88 (R75 Integrator) pr 418-018:PAGE 1-303 i rms N ee i ey EE A | A $2 63 6%6%60ET 6M6M 7070 IX 1% 1%IW 1m 7m 1a 1m | Primedica-Worcester Project Number: PABX-BBA 418.018:PAGE 1-304 Page 118 PeakArea Report o NamSounrpeeno:A00sSA00TUP0L4E ovecSrrs 501654 Gongess Gommpum or Ima mmm Se a tn tn PrimediaWorcester Projst Number PABXCBBA QI | GESgF--T----g--JE Be ARETE a LTT DGornun. or sper 418-018:PAGE 1-305 Page 1o i RT EE @T oer EE Te e SaaS eE a TTR aR ae vk aw an em am im ik TR ihre ie ie in ve me) Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-306 Page 120 Peak Area Report NotboSnuknpRleoraems::F05A5H1SEOAU5L0E1 vtSecr: 01 713 tet aeasm ABSA ro an " compaess +o cwmimpeium Imbel HowOMIhEe SMR omtt at tno rd rg Primedica WorcesterProject Number: PABX-BBA 418-018:PAGE 1-307 Page121 fsBecpeaeperite 1Lo0y0aBaRmmRmRaREBL RGETIOROSSiiHa.48 QL ET oma, en to sro fo I RE Te TTTTe RRR Frappr 0 -- ome NE NEET = Re Te RTO eTrhe e Te Te e a e ctor mana on 1 see 20 rrr PrimediaWorseProj Nb: PAB BBA rage 12 418-018:PAGE 1-308 FO PeakAra Report NotebookReference:PABK584-1047 veges `Operon: JO vc. A... 7 Zz SU EE Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-309 Page 123 Buca File| 01\ISD DATA\PADLTABLBBRVL-047-1\04TIO6L 8_ ink: 63 RE aiden fie QE rE cos pnsx_swaw (7 tnceseator) re aRTVTeReh eRTT Th 7k he TeTR ili ener, r rr EEETNhNE T E REEa TTR ev va e eh verhiTRl7 evk Th vk Tie 3kRT Th Primedica-Worcester Project Number: PABX-BBA. 418-018:PAGE 1310 Page 124 PeakArea Report [IcSamidaimc:tiGoRn:OrUePdE onecpedn]: 1301758 IaS ABSASnpasesxanron mare6 Gets cometens TBR fm tame tems Primedica-Worcester Project Number: PABX-BBA 418-018:PA3G1E1 Page 125 Saa DaEtaRFileh iD:dD DBeAIEAn\PAB\BARK-88R\1-0UT-\GHTIORE Sf5pi_esrnViiaalr: 62 QL SRT BeeShove mae wen (x7 tnceszaser) So 0 GGR e 6% R enek aw7hTw 77%Th Th Te 1hTh Te wa eltrey S90 FEAT RS TR TE Te an Tht RT TR ve vn ee Th Te Th TT Te e Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1312 Page 126 Peak Area Report NosScaomtplReeaamees:o3o3oGnURL7 oucSnm: von 832 [ounI ioAahem:S ooroi,ey30SATAPABPARKBBA 07. naarnmVaaann 6:E5 r E 2 ew= s Ta ow wm Toe oa oma gnre hg. rimeticsWorcetr Projct Number: PABXBBA 418-018:PAGE 1-313 fr riarr Br er TH ETT Efrat,son es sven I o E e E Te RTR E TERT wd TR aneh il Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1314 Page 128 Peak Area Report o NonSeurRplomo:sF0B3L8S5O7N00094450 onecpaecs 01832 ee ATA SUE SAAABLOG1I1,var 4 [ER -- Primedica-WorcesterProject Number: PABX-BBA secon pore sets BgEnEEi a 9T9D r P EeTSTEMnIerE , EET ET coon sn ee censor 418-018:PAGE 1-315 Page129 fens I EETR TRTRTER hee ee TR, TahThRR @ Aon: 338 Re) TTT TT mm ee1 TR evama ikeih 7h Te ThTe ih TRTR Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-316 Page 130 Peak Area Report NoksSooarmep eamse: sssSOAOUoP T [ Soa fmgest | Gmmplem J Imfl ln teime ties Primedica-Worcstr rest Number; PABX-BBA 418-018:PAGE1-317 Page 131 aLErsag95 3ISpRDp EA T E5 Eie Qho TE BRoorE n poAmapas ; | oe TRRT TT eT TR Nh rye eet noe o E a EE eeE jBTe a ve TMT E Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-318 Page 132 Peak Area Report Y Nese CSR oT Or smr eoa svai nr e:FaAXit .t gNe Ns R ---- es2 r cowmiipre SR ow roEnmg cpp Ta heintk gt ord tr PrimedicaWrest rjc Number: PABXBBA 418-018:PAGE 1-319 Fage1ss LE LT pe---------- ag IER amen TE er OTT ST os. sept i EE ETE ERR EEE EEE T EEEe EETSRE T Ee TEETe ETEE S CEE h Eh hE he ee ns mp0 mo vs primed Worcester Proje Number: PAB BBA 418-018:PAGE 1-320 Page134 Pk ren oor 0 EmE emN orovisgpma BIeS nasonn snRERES 77 eran EE op pr rres Prmedica Worcs Pojct Number: PABX-BBA 418-018:PAGE 1-321 Page 135 quie ou stsce moon5 tr = 10956-GROUP FO TE EE LT mongon sm sve fee a TR Frere8 em EE BR Te TE ee EEE ReaRaah na Primed Worst rjc Number: PABXBA 418-018:PAGE 1-322 Page 136 PR rene 7 oakrea Rept ggee EIRE n, enSRE Eula o som in ss Primedica:Worcester Project Number: PABX.BBA 418-018:PAGE 1-323 Page 137 Sesepe BEiv BERA L67[=8 QL BRE comma, sun at scracon pu]| ! | a EETR RR eR TE Te i 4X4 6a 636% 610 6m 6m 100100 TX IW Te 1M Tw = IR 1m Iw T R e e E ew ahae amew am 7m ir w tm tw Tw ik tm im re om Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-324 Page 138 Peak Area Report NosSaempleanme::P02A858B00o71r1]50 ecu: 2m 1011 eer TESESoA01 1, ware3 GompHeds omwpiislem leWfo) WBn umam hambe 1 cernstrr gnwsper gn Primdica-Worcester Projet Number: PABX-BBA wm sto; He Ens BE OT EE wrengon ot spn 418-018:PAGE 1-325 Page 139 - rr a a IR ak ew ew ew em ak ew Tm veim 7%wie 78 3% ve te TTR Wh swew ew ew ek sw 7m ie sm 7% 7a 1s. ve 7% 7h 1m Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-326 Page 140 Peak Area Report NetbSoaomkpeleinamee,:F10A9C1B.5n9000P81150 ove Apcuera 13011031 Sent pogtams ot 1 oe ei Dope) wean baigge gn rd rg. PrimediaWorst Poe Nbr: PABXBBA no 50 s rse " nty-- QT EEE oceans ce sncsecen = 418-018:PAGE 1-327 Pager EEE ET RR B e a R e A TE EI Jar OI IC I EE MET ME MEDIE EM T Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-328 Page 142 PeakArea Report NekehoSskFmeerree:P2A7G0E0S001047 onAecqnsns 13011050 sumanounoFFieosaamm.oSArShH,S8D40ATAPABPABCOBA O71 srm Vritrc o:t173 Comune 2 Comopiums wis In mm Pum Putin wow a J PrimediaWorcester Projct Number: PABX-BBA rie 0 sae eet: ET ETT means te errs 418-018:PAGE 1-329 Page 143 T TT Teh ER TET eR py $e Ee ET T h rEe IT Th o | TTR kak ae k ehr eke es 7h1 7%e 1%e re 7k ie im 3m Ttme rimeticaWorcesterProject Number: PABBA 418-018:PAGE 1-330 ge144 Pksven Bape PR J otte ve i T = EEE PitsNoss Fo Nab FAR 35 _ 418-018:PAGE 1-331 BET desea Te wr QT ERAT Daraui cossa aa _sn ea. (RTE Integrator) z rT h h eer yy. ow ---- IE | Le enss : i 6m 60 6eM w 70 710 73 13 7a 1%7 1% 180780 Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-332 Page 146 Peak Area Report NowbSanakmFpalrees:15C8PO0LUS Po1b150 Owe AEec1e291 1s30 Gomsapmit Comwmipteans Teaelw Mam m Euthmpee Tn omns a gen sperma rp Primedica-Worceste Project Number: PABX-BBA 418018:PAGE 1333 Page 147 Sac ile 3100 AP PABBA 00OUIITED Viad 72 SEE deen TE er OE STEvo wt meron ee OREREeh arr a em JETSIIA EM MEE amen mes me se 10 15i0mes 2000 nes primedicsWorcester Projst Number: PABYCBBA 418-018:PAGE 1-334 ws Foksma aren 0 m JE E JO auIrMEI ns co NsoeO rss moIar peepaee EEal Tas nage pe Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-335 Page 149 aoHleeie VliEdSaSmNaA MRSOIRTTuerlTtta:3 SE EIT an 7 separ I" I TT Te TTRT TRATR i -- tive Wk w isyR eTh Thi 7a 7 7h Th Th th S EEEh RR Primedica-WPororjeccetNsumtbeerr: PABX-BBA 418-018:PAGE1-336 Page 150 PeakArea Report NotaSampReatmteA1635S50A0081]150 oeAoceas:a 1210 fmspelt | mgrgums Impal f MmE Bie A -- Primedica-PWroojertNsuembiesr:PABXBBA FHee iopeammney fweut QL EEE enon an re pan 418-018:PAGE 1-337 Page 151 Coie 6 4B 4% Ge 6%G0 enew ew 20 70 10 TW141%Te1% 00 TE i T o I a T @i } | R A T Tee Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-338 Page 152 Peak Area Report NoneSunRpieenanee:: PcA3BeGARAO2U4P71510 owecEauos 01 1220 ernER BASAPNA11, re ro fomgpess comme Imps! em times t csrc gso drtd. Primedica-WorcesterProjectNumber: PABX-BBA 418-018:PAGE 1-339 Page 153 Sn {A HEN delet Ti To ver TT dioEnNTE avons,an w5eeonegreenes 2 | " TE a T EEJEN EEE TERE RE Maron I oo a MN A TER enh we in sk ae ve 7h Th 7h 7a 7h Th Th ihel connor wanawn ma tens rs Primstica Worst Projsct Nbr: PABY.BBA Pos rea Bagot 0 IJ EE-- E 418-018:PAGE 1-340 pope 190 aFoa r I Tw wR Primedia:WorcesterProject Number: PABXBBA WATLP SEr steinh apo evEeles BF IRE Tr QR DT faa,san. wre epee 418-018:PAGE 1-341 Page 15s i TE TeTee TETeTeTE ERT lg a e leree TEE ee TTReTh rimediceWorcs Project Number: PABX.BBA 418-018:PAGE 1-342 Fae 156 peensRepkort Ip m-- m:-- miFgensoni ee ESTs wenRATES po7ns og=ee E=D1=m2ime mye re mm tetop eet et Primedics-Worcester Project Number PABX-BBA 418-018:PAGE 1-343 Pogo 157 J in + sym Ep------T J 7 QL BRE omnes x 1 saan) i TEE Ey ee Bh OEE wor ETR a Te eT TTveR ve ew an aw aw vk 7h Te Te 7% ik Tn 7h Th Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-344. Page 158 Peak Area Report mgt scrou1 ounpt 12011330 Comp2nst owmsapsitem DTafeol m maumei mene Primedica-Worestr Project Number: PABX.BBA sm unt S eIEEE QE smon rs 418-018:PAGE 1-345 Page 159 1 EEEo EEREh TERE EERE RET a ee on B R r RT L Ea T EN E TTT Pimetica-Worcse rjc Nahr: PABX.BDA PekAra apo 0 SerHranEEsm 418-018:PAGE 1-346 Page 160 eswem or wmnte comme SD nme spp ei------_------------ Primedica- Worcester Project Number PABX-BBA DBEynEepie + L5opnmEppE hguE PE IS ann go ih opens 418-018:PAGE 1-347 Page 161 - o A l a TIER ve sk seE em E kiR TR7E7m TRR 6Rme veR 1m thR me TR wh eE e akeeem em en tw 7 vk Tw te 1m ve 1h Em | imeiWorcstr Project Nombsr: PABBEA 418-018:PAGE 1-348 ge 162 Pesesnsrape 0 mE gg aea eres beSirspiasiecemnican, erm oom comes EE nme mene omee erect SA TR SAAT cen oo EERE et @ ET ARSENY METHODS PARX_BBA.N (RTE Integrator) 418-018:PAGE 1-349 Ss Fr re E eR Fr ee eB ee i A| PrimediaWorcester Proj amber: PABX-58A ose 166 418-018:PAGE 1-350 PeakAres Report 0 Rs ENrEe E SaT. Or CA. resus ---- Primedica-Worcester Project Number: PABX-BBA ie sSe E nl 11036 GRovRLIF a Te wer 418-018:PAGE 1-351 Page 165 J E-" EE EEE AEEE EEE i ---- eer AMea Err i Then ewen ew an em sw 7k vw 1k Te 1a 7% tie = y th 77 h m) EET RR RR rimdicWorsen Project Number: PARK.BBA 418-018:PAGE I-352 Page 165 PO I sk rea Bape ru ----- JR -- o rimedicn-Worcester Projet Number PABX.BBA coining psn Lo 5 TI ST eros,sean oe eens 418-018:PAGE 1-353 Page167 L E oa EJ R N AETEE EE emerPS Boe ha EE ER TIE awe e wh ve en ve ih Te Th Th a ie Th ve 7h ih re PrimsticaWorcs Prject Nbr: PABX.BBA 418-018:PAGE 1-354 Page 165 Peak ren event 0 SsHr EE". eLro o Jo OI... oA --------------------_, Primedica-Worcester Project Number: PABX-BBA enTie O5EsEmAonBL Qn ERS]Dainoos pass seu (575 tegeator) A150IPAGE 1355 Page 169 eel TT RT a RT RR RR a) PrimiWorcs ProjectNumber:PABXBBA A180 PAGE L356 age 170 oareaRekport PO ren SchI wT nI acss wad com3es coHmp SwEDR mm ages Primedica-WProojrecct Neusmbteer: PABX-BBA 418-018:PAGE 1-357 Page 171 sSE aaapir e im 0 e a T BOONEMBrvss 36 TER ERTDens hn sees 4 || _ D TR aVe d one Tn T Th Tra 1e h J i TR Ta gy meen e ES R ehE Th me "rei PMs rt ng AA To Ww we aw AT SR am TN_J0 18 7H 7a1 te Thim mh seen vargas hao 1 Teen os wr Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-358 Page 172 Poak Area Report NobSoamFpelnee:s2PxACBONBTARO5L67 wehSerres 1301 1540 Gomest 2 Comeptes wor InBh wm mame Pte fame Primedics-Worcester Project Number; PABX-BBA 418.018:PAGE 1359 Page 173 Boararite omssEBRB ARR TET0HCvin 17 MET ET oar gos sweets = w| | | REE var wa RaTTR TR RTE TR TTR pe TE TTT T A a ee N RRR R R i Th h 77e TR 7h th Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-360 Page 174 Peak Area Report NotSteanFseannne:2F5S01A1087 oweASca e125 160 cempens conmpiam lmfal eume tebe Primedica-Worcester Project Number: PABX-BBA amr nso Bg RT Lo PE 418-018:PAGE 1-361 Page 175 3 EEEA TAEETERE EE EERE i ---- FRemRREE RR rw rindi rjc Nob: PAB BBA 418-018:PAGE 1-362 _-- es ra Report 0 IsEonEoE: [EsTm Ce era. DMSO OATAPABPASK BOAT. stemnoe o1 InstrumentMetnodName: PABX_BBA mr comer come ZED mame mane Primedica-Worcester Project Number: PABX-BBA gage 53 mFpR BE RY ni avn-- 3g we LT ET eons ws ees 418-018:PAGE 1-363 Page 177 Ls 618 4B 46 GH SH SRAMGh 4% 16 TT 1%TR HTh 7 1% 78 Th EE TTTe Te Te Th TTa \ | T ee T Te re mses | PrimediaWorcesr Project Number: PABX-BBA 4IBOISPAGE 1364 ope 78 [-- 0 EJR EE or genpyisstion A amcsassane verRE InstaumentMethodName: PABX_BBA w= re etreem PrimdicsWorse Prt Number: PABX.BBA priEe + E 33EmRma noonfort 23 @r EHngR Ho0S PAs S _saA. 0 (RIE Inteseacor) 418-018:PAGE 1-365 Page 179 o Te E Te TR TE Te TR er Bt e a ee CTR a ae ae se en ee sw te rw tm 1k a 1%1m te wm 1m | a ter RRR oo a] T E TTE E T R E RT RETE E e E R e rimetica-Worcs Project Nbr: PABYCBBA 418-018:PAGE 1-366 Fag 180 Parken opr IEpEET ... [rEo T CRISIS pn, son STE coI r copmyps SE E E sam eg FO --. aren a PrimiWorst Pret Numba: PABXBBA I Ip 5 LE ETen. re cars 418-018:PAGE 1-367 spre ore Le ai eRm ah eGrseGnWce4HerGFe Et 4k.4% 1 rTWeTnEe7e%Te TR 78I% TreHse0 m Ce Th ee aa n akw am am sw ti m s im iw Te 1 rm im vmm e primiWass Poet Number: PAB 3B 418-018:PAGE 1-368 J ak ve pont 0 IEarEpa csrlrn i -- comes ESE wp mums myn PrimetcaVorssts Proj Nas: PAY884 cies os et Bp lt QE I cen, en ae napa 418-018:PAGE 1-369 rages : A EEE RRR gg oe 50 a A TR E RR Primedica-Worcester Project Number: PABX-BBA 418-018:PAGE 1-370 Page 184 Peak Area Report NotSaamspRleeannee:2A5G70S600167 oweAScqrua 25 748 Comp2ndt | Gowmistptms lomfo) wRm meume mime Tr or cr otmr gn rd n rg rimeWorst Proj Nb: PAB BBA cio pr es Rises ful?) TE ETE es. er er 418-018:PAGE 1-371 Togo 185 > TE EEE AEE ET EREEERE EEE -- E i E E mE E NN E Primedica-Worcester Project Number: PABX-BBA Peak ra epon ITsEr.e.. 418-018:PAGE 1-372 Page 186 sieg rimeticaWorcs Prjst Number: PABX-DBA ane st Amt Em pin TTEET fs one re nes 418-018:PAGE 1-373 page 137 - TT TR REe m Ot a TR ER ta we en ik am vk 1% vk ve ve 1h te im im 1% 418.018:PAGE 1-374 3M Medical Department Study: T-6295.25 3M Medical Department Stuy: T-6295.25 Analytical Report: FACT-TOX-169 LRN-EO1-0150 Analytical Fepor: FLAACNT-T0O1X--0116890 Study Title DeitnerthmeinMaetviaolnoonficthAeciPdr/eCsheolnecsetearnodl CCohnaclelnetnrgaetiaonndoNf OPEerLflIunovresotoicgtaatnieonsuSftoundaytein(RPaFtOs.S) Analytical Laboratory Report Title Determination of the PresFelnucoeroacnhedmCiocnacleinntrRaattiSonoroaf SPaermfpllueosrooctanesulfonate (PFOS) Data Requirement Not Applicable Author 3M Environmental Laboratory `Study Completion Date July 06, 2001 Performing Laboratories Sera Analyses Sera Extractions aM Environmental Laboratory Pace Analyical Services nc.--Tier2 Buiking 236-09, 35 Bush Avene St.Paul MN 55106 170M0inEnemapSoliirs.e SMEN.5S5u41e4100 Project Identification 3M Med`iAcraglusDIenp-aLirftemSetnutdyS:tu4d1y8:-T0-168295.25 `3MAnLaalbytoircaatlorRyepoRretq:ueFsAtCNTo.TOEX0-11-061880 Total Number of Pages 68 34 Environmental Laboratory 331 Environmental Laboratory Page| Page | 418-018:PAGE 1-375. 3M Medical Departmen Study: T-6295.25 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-EOI-0150 Anaiyical Report: FLAACNT-T0O1X.-0116890 "This page has been reserved for specific country requi-ements. 30Environmental Laboratory SM Environmental Laboratory Page2 Page? 418.018:PAGE 1376 3M Medical Deparment Study: T-6295.25 SM Medical Department Stay: 6265.25 GLP Compliance Statement Analytical Report: FALRCNT--ETOO1X--0116590 Analytical Report: F(AACNTgoTrO.Xo-t1e609 Analytical LaborofatPoerryluRoerpooorcttTainloe:su_ifDoentaetremi(nPaFtOiSon) oFflutoheroPcrhoemsiecnacleiannRdatCoSnecreantration Samples Study Identification Numbers: T-6265.25, FACT TOX-169, LAN-E01-0160 TAwidhlmihisnhsitseutrdeayxtcwieoapnsti(coFonDnsAd)iuncGttoheoeddbiunLlalcbeootmerpdaltliosartnycbePelrowawic:tthicUeni(tGeLdP)SaRteeguslaFtoioodnsan21d DCrFuRg Part 58, Excoptions to GLP compliance: + Tfohretrheewaenraelytiwcaolspthuadsyei.rTohtoerns ffor tsitsudsytupdhya;sneawafos ctohnesindefredpthoaesnedanatdtohneo Cgeonnesriadtoiroendatnodstsarthattih oprfesmcpaeectiomnfetnthse.seThsepeacnialmyetnicsalfosrtaunddylypshsa.seSiwnacse the `etxepcehncitceadl pferrofmotrhmisanecxecaopfieoanc.h phase was onrly separate. no afect = + Automated data colection systams have not been valdated as roquied daactcaording to 21 CFR 58.130(c). The hard copy printoutis considered the raw + dTehtersmtianbeld.y ofthe reference standard and internal stand wera not Deanna J. og f, M.S., Study Director Mote 16 Marvin T. Case, D.V.M., Ph.D., Sponsor Representative Date Duns 200 Date KisfteindJ.eHanseLnb, Ph.D.. PrincipalAnalyical vesfgaior rDaete Cow) Wiia am K. Feagen, Ph.0., Laboratory Manager 3M Environmental Laboratory $M Ewironmental Laboratory CTaatodstor Pages Pages 54 tit Ds St TTS scat epamarscy Tzss 418-018:PAGE 1-377 FIp--d ct eg AwGTTs 0109 GLP Study--Quality Assurance Statement vaya LaorsayFopon Toa: CotofmheaPrsannce and Cneansaton Soros of Perfluorooctanesulfonate (PFOS) Fiuorochemical in Rat Sera Sy Wasson mbes: A285.25, FACTTCX165 LANEDL NSD re -- rr ~~ [motores [mee ER EE EE [ae oa m ||oeo enm s | [sawmoer | [aewmwo |_wwoonses|orsoesmio| r eas sao Fm baz goier Erion iaotars cay ran pages 4I8-0I8:PAGE 1-378 3M Medical Department Study: T-6295.25 aM Medical Dopariment Stay: T-6295.25 Analytical Report: FACT-TOX-169 LRN-E01-0150 Analytical Fepert FLAACNT-T0O1X.-0116890 Table of Contents GLP Compliance S1Q1OMeNt... mm--0 (GLP StUy--QUaHY ASSUAGE SIABMON rd Sly PISO ANCOMBOS... T I010GUCHON 81S PUPOSO...vv A Specimen Colecion and Aas enmm------ Specimen RECeipt and MaIENANCS vd Chemical Characotf eherRefierzenacetSUBiSIoANnCS vr 10 MetFhTod SUMMaAeS..........A..c-- . smmnemma-------------- ----)1" L J S ------ -- nni-- m--_| De O Rrrpmmmome mm---------------- " wem-- Data QualityOBeCHVES and DATE INMGQTRY rrr 13 DatSa Suummmamroyf,aQAUrnaallytyyseCs,onatn]d RAENSBUIS.Y.S.ES RESUS... rrr 1134 SStuamtoofefSmDaammtpaaelQeUnrRBeItsYyuis......v..v.......... wem--m ------ Statistical Methods and CIGURIIONS crv 15 SHaleMent of CONGHSION.....crrr onsen Vl AppendiAx:Control Matrices and Test AC... 16 ADPEBN:PIOI0GO! ANG DEVBUONS...rs Appendix G: Extraction and Anytcal MBINOG re 30 EHTPSL.C8..E4l,eEcxttorsapctriaoyn/MofasFsuoSrpocehcamtiocmaletC(oy1m,6poPuagnedss)f.r.o.m Serum for Analysis Using 31 ESTeSr-u8m:5E,xtArnaacltyssiUssionfgPoHtPaLsCs-iEulmecPterroflssporraoyoMcatasnsesSupiefcotniaotmoeotry,Ot(h1e1rpFaigueos)r.o.c.h.e.m.i.c.a.l..s in 46 J ------------ ADPONGIK E: Da SPIBUSNERIS. rrr: 6 J-- Appendix G: Interim Certicate of AnaySis........ enn 68 `AppendiHx: Report SIgnalurePage... ao 68 a EnvironmentalLaboratory $3 Environmental Laborators Pages Pages 418-018:PAGE 1-379 3M Medical Department Study: T-6295.25 3M Medical Department Study: 6295.25 List of Tables Analytical Report: FACT-TOX-169 LRN-EOI-0180 Analytical Report: FLARCNT-T0O1X.-0116890 `Tabl1e. FemaleRatFO Population DemografpohrSitcudsy (Argus 418:018)...........8 Table 2. Characterization of the Analytical Reference Substance in Study FACT TOXTable 3. Target lonsMonitoredin 3M Laboratory ANBYSS.........sumnrn 12 Table4. Determinatioonfs the LOQ in the Analyses of PFOS EXU8EIS rcv. 14 `Table 5.FCAhaCrTacTteOrXi-za1t8i9o.n.o.fctrheesCosnntmroslsMrastsrmiceenssUmsmesdssforrmSmesrnaiAsnalyses in Study 8 Table 6. Characterization of theTest Articl in Study FACT TOX-169 errr 16 `Table 7. TOX 169 Data Summary of PFOS Concentration--Rat Serum (g/mL).......... 57 3M Environmental Laboratory SM Environmental Laboratory Pages. Page 6 3M Medical Department Study: T-6295.25 3M Madical Department Study: T-6295.25 Study Personnel and Contributors SDteuadnynaDiJr.ecLtuoerbker, M.S. B3uMilCdoirnpgo2r2at0e2T6o0xi2cology (Medical Department) St Paul, MN, 55144 Analytical Chemistry Laboratories KaSreMirsaEtneAnvniaJ.rloyHnsamenessnetna,l PLha.bDo.raPtroirnyc(i3pMalALnaabl)ytical Investigator 418-018:PAGE 1-380 Analytical Report: FACT-TOX-169 LRN-E0I-0180 Analyical Report: FLAACNT-T0O1X.-0116890 S3pMonCosroprorate Toxicology 3BuMikMiendgic2a2l0D-e2pE-a0r2tment SMta. nPianulT,. MCNa,se5,51D4V:M. Ph.D., Sponsor Representative PSearcaeEAxntarlaycttiicoansServ.cas, Inc. Tier2 Faclty 3M Lab Contributing Personnel RLihsoanAd.aCDliocmk"en MHaarroilednO,M.JoHhenyisnogn" KBoolblWJ.. (WKyunenhiewein) Swartout" Conta pretesse t rion Location of Archives ASLalpbeoocrriiagtmioenrnayls. rpTaewhrtedaaittneais,ntgparrttooitctolhceoolaa,nndaalnaydntaialcnyaatlliycptahilacarslaafrooefrpootnrhctiehssattvuaednydbaaerredon raaerrsccehhriivvveeeddsaaatmtpttlhheeesa,3MMasEEnwnveviilrroaonsnmmetenhnettaall Ltaesbtoirnagtofracyi.ltRye(sAerrgvuse)s.amples of the control article and vehicle components are stored al the a Environmental Laboratory 3M Ensironmental Laboratory Page" Page? 418-018:PAGE 1-381 3M Medial Deparment Sud: T-6295.25 SM Medial Doarimos Sty 629525 Anaya Report: FLARCTn-ET0O1X0-118609 Avant Report FeACTsT01o69 Introduction and Purpose pEThoeertbrpomurirsnpooacstaeonnoefsotufhlehoesntaPutrdsyes(sePnoFcOedSe)atneidrmrCitonngschooensapairmeopsnleenoscfekPaaenrnduocitoanaocccecanonrtedacaiunotcneoonfwaithe h(ePR3OS)pr1ottoncoel, Movant: AGeLCahGsloagtorlDraEnge andrNOeELSIonvepsgaatgon SousaShn aTts:TFoenmaSleFTOS mEwea0rte5mcaoGllolmaeecnstd,edf2ofourlweaalnlailayossitosofbattohsaprceocdimofiieindgisntteharvPaloRs. SoTfhme0evicnamtlesonentniovcfeadthcfeeostreuccliyhvewesalst(etVroO)dLew)t.earnmTihnwaeihsthte y Vas iiaed 31 Jansary 2001. TToeissrtoSSnyosgtlroeumops pof feImatlewaarse Gwoern uotshdaaTs ahente8st,smystoomv. Garoau0p.1,oGfrXoRuGps2tearndl Gornoy:up & SSGroronuidposbt3og.i0mCaiornmdn45a5l1Goa5nyswwiponriermemvientlaeotnrieaocddnP,orFaSOnhSdldecososnaetcsi.cniFn0ga.h5m%oauTgwrbhaetdhwaeayrb2o(0vo!esnprrhelsuymteoidt er mGeasttaonha{tCdoamaotndesbeeconogrr)ouop)ddaay42p4oOfsparreusummcedaeutsataGrav(eartsaee8y.44 1 Panral The test systam species and strain selected was the Ci:CD'(SD) GS BR VAF/Plus rat received oituenrtmsedof 5npgaHrorfsMoone,Tabblta w1a:ts nt meal 120360, av 1 PramOrOrs ware valalea from Charles River Laboratories, Inc., permanently identified using a Mone!" self-piercing ear tag. Only FO generation rats were identified with ear tags. F1 generation rats were identified and Table 1. Female Rat FO Population ---- DeSomoiongeisrotanephifcosr Study (Argus 418-018) oo [i[mmeVr so osconcscerr| |c2o0| ]miteninoeeassgss ] Geoaose =T Ts a| Tsomnoosgrrrrrooss..nmeemosncseeess CT TyS-- ---- TT cis oommeer omes T1w |1cnoraocarammrourgogrrrsroocss mee|n ooewsen T [a a] ] r onorarros Gcemeen TTa hT | remme gres s| ow Ermer Laboratory Som atin rages Pages 3M Medical Department Study: T-6295.25 3M Medical Department Stuy: T-6205.25 418-018:PAGE 1-382 Analytical Report: FACT-TOX-169 LRN-EOI-0180 `Analytical Report: FLAACNT-T0O1X--0116890 `Specimen Collection and Analysis pInootlheedarnaalyttiecralsswtourdey sreenptorotedthhoe3reM, 2E3nv5isreornamesnpteacliLmaebnosractoolrlyecatenddfarnoamlyfzeemdalfeorFPOFrOaSts.aTnhdeF pnruemsbeenrteodf bseelroaws.pecimens collected for analyses in the analytical phaseof tis study are `SSepreacSipmeencsiCmoelnlsec--t5e4dsfpreocmiSmteundsy(FGOroFuempasle1st,hGrDo2ug1h, 1C3a:esarean section) SSeerraa SSppeecciimmeennss----7584 ssppoecciimmeennss ((FF1O fpoomoalleeds,itdera, yG5Do2f1,lacCtaatoisona,renaatnursaolcdteolni)vory) Sera Specimens--49 specimens (F1 pooled liter, day 5 of lactation, natural delivery) BElnovoirdosnpmeenctiamlenLsabwoerraetocreyntfrriofzuegend.andSeornumdrywaicse.then harvested and shipped to the 3M `mSeetrhaysia-emrptluestywereteheerxt(rVaIcBtEe)d.bSeagimnpnliengexotnra0c5tsFweebrreuaarnyal2y0z0e1duussiinnggahnigiho-nppraeisrsiunrgerieqaugiednt and crhersopmoantsoegrmaopnihtyo-reilnegctmroodsep.raPyfFtOaSndleomvemlas swsersepeqcutarnoimieattreyd(bHyPeLxCt-eEmSalMScaMlSib)raitniotnhevemrulstuisplaen extracted curve. Analytical details are included i this report `Specimen ReceiptandMaintenance `LTahbe.3MAllEsnpveircoinmmeennstawlerLoabroercaatlovreydrfercoezievnedinrgatoosdercuomndsiptieocnimoenndsrycoilcleecatnedd waterAergiumsmeRdeisaetaerlcyh apancskfse.rArfetdertoexsttroarcatgioenaatt-T2e0r2C, t1h0eCs.peScpiemceinmseannsdwtehreeetxrtarnasctpsorwteerdetotPuarcee dTiecro2ldfroonzeinceopnacikces:. c`Coomnmtreorlcimaatlrsicoeusrcuessedanidn aserreaparneasleynsteesd pinerAfpopremneddixduAri(ngseTOTXa-b1le695).weSraempoblteasinaendalfyrzoemd at the 3laMboErnavtoirryonamte2nt0alCL+a1b0orCa.tory wil be maintained fora period of 10 years and will bo stored at the 3M Environmental Laboratory $M Ensironmental Laboratory Pages Page coat i T35 oar 418-018:PAGE 1-383 re Chemical Characterization otoeahne ete of the Reference Standard mr [Teme| ee Fa. CharcifrzhartaoinlRefsnesubsanenSFACTTOK169 1 PL rowsa THPFOS (Tetrahydroderfluorooctanesulfonic acid) Tes [Epmonows| [rege Conations| wor Fraznst0c | | 2005 20C rerimein| iscsipe] pone ew 1e Eet woe |wr] 418-018:PAGE 1-384 3M Medical Department Study: T-6295.25 3M Medical Department Stuy: T-6205.25 Analytical Report: FACT-TOX-169 LRN-EO1-0180 Analyical Report: FLAACNT-T0O1X--0116890 Method Summaries Followingis a brief description of the methods used during tis analytical study by the 3M AEpnpviernodnimxenC.tal Laboratory. Detailed descriptions of the methods used n this study are located in 3M Environmental Laboratory "PREPARATORY METHOD + ETS-84.2, "Extraction of Fluorochermical Compounds from Serum for Analysis using HPLC- Electrospray/Mass Spectrometry" Apaniarliyntgicraleasgeerntumwasasmapdldeesdwetorea elaxbtorraacttoerdyussainmgplaenainond-ptahierianngaleyxtter-aicotniopnaiprrowcaesduprea.riAionnieodniEnatcohMIeBxEtr.aTctheed MlaIbBoEraetxotrryacstamwpalsetwhaens rreecmoonvsteidtuatnedd pinut1o.0ntmoLaonfimteratgheannoelvaapnodraftiotreruendti dry. through a 3 mL plastic syringe attached to a 0.2 um nylon fier into glass autovials. AnaLyncat Memon + ETS-8:5.2, "Analysis of Potassium Perfiuorooctane-sulfonate or other Fluorochermicals in `Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" The analyses were performed by monitoring one product ion selected from a single primary mfoonlecchaurlaacrteiroinst4i9c9,ofsealpeacrtteidcualsartfhleuoprroicmhaermyiicoanl fuosriPngFOHSPL(CC-sFESSMOSxH)S.anaFloyrsiesx,amwpasl.e, qfuraangtmietanttievde atonaplrysoidsu.ce ion 98 (FSO). The characteristic ion 99 wes monitored for 30 Environmental Laboratory SM Environmental Laboratory Page 11 Page 11 418-018:PAGE 1-385 SACS Sh TOES tet epmanSo T2525 Ars Report: FpACTtIONi160 AAcI k SE To oli present of stig uss un thenayphase of iuy. Liquid Chromatograph: Hewlett-Packard" Series 1100 Liquid Chromatograph system on ppEEBEleaeooa ,emReYaemnnma Analytical column: Keystone BetasiTM Cy, 2x50 mm (5 ym) e EEMasrsyiSpenA ce tnrometer;McHrromass APUMass Spectrometer Quattro I" Triple Quadrupole system [artei mipteoSdasseonaoisr J --_ [rr o|mT saimni s |_romweoicnion pros ws ws mma pagers 3M Medical Department Study: T-6295.25 0 Meal Deparment Stuy: T-6295.25 418-018:PAGE 1-386 Analytical Report: FLARCNT--ETOOLX--0116890 Anayical Report: F(ARGiT.gT0OrX.-0116890 Deviations Deviations from the original protocol and methods are included in the Append B. Data Quality Objectives and Data Integrity The following data quality objectives (DQOs) were indicated in the protocol for this study: + wLeiingehartiitnyg:. The coefficienotf determination 1) equal 0 or greater han 0.990 using 1x + cLailmiibtrsatoifonQucaurnvtei,tadtaifoinne(dLaOsGt:heThleowLesOtGstiasnedqauradltthoathies blootwhesttwaoctcismpeasbtlheesmaatnrdiaxrbdlainnkthaend is calculated within 30% of the expected concentration. + SAtcacnedpatradbs,leprPorveicdiesdiotnh:eyQuaarleitqyuaconnittraotlesdawmiptlhienstahreesreelqeucitreeddctaolimbersattio2n57r%anpgree.cision + Confirmatory Methods: If a confirmatory methodisused, an amendment o this protocol should be wien + Dreetmeonntisotnriatmieon(aopfprSopxeicmiafticeiltyy7:.P3FmOinSutiedse)n,tibfiycatthieonchwairlacbteersiusbtisctapnriiimaatreydfboync(h4r9o8m)aatnodgrtahpehic characteristic product on (99). Data Summary, Analyses, and Results DLaabtoarqautaolriytyporobtjoeccotlivoers fFoAr CthTeTaOnaXl-yt1i6c3al(psheeasAeppofentdhiisxs8t)udwyerouetmineetdwiinththteheIeMxcEenpvtiiroonnsmennottaeld in this report. a EnvironmentalLaboratery 3M Ensironmental Laboratory Page 13 Page 13 418-018:PAGE 1-387 ssa JR--w-- e Summary ofGulGonAnsys Ress Linearity: The coefficient of determination (2) of the standard curve was 0.990. a Suma 2 geeas eben bey Methods). Limitsof Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the rn Ta ETCH TOL rie + Internal Standards: The internal standard (THPFOS) was added to all samples and statement of ata Gust [P-- rare 3M Medical Department Study: T-6295.25 314 Medical Department Study: T-6285.25 418-018:PAGE 1-388 Analytical Report: FACT-TOX-169 LRN-EO1-0180 Analytical Report: FLAACNT-ET0O1X.-0116890 SummaryofSample Results PFOS results have been corracted for purity of the analytical reference material. + cSoanmtrpolleasnifmraolms.CTohntersoellAenvielmsalwse:reLosiwgnliefvieclanstolfy PloFwOerStwhearnethooftseenfdoeutnedctinedthien tlhoewsdeorsaeotfestthe animals. + `SaanmimpallessinfcrroemasDeodsweidthAdnoismeallsev:el.InDgeetnaeirlaeld, sPaFmOpSleledvaetlastfaobulneds ainrethperesseernatoefdtihneAtpepstendices DandE. Statistical Methods and Calculations SAtpaptiesntdiciaxl mFeftorhoedxsamwpelreeclailmciutleadttoiotnsheuscealdcutioagteionneroaftmeetahnessearnudmsstaanmdalredadtevaiaitnioFnAs.CTSeTeOX168. Statement of Conclusion UofndalelrFtOhgeecnoenrdaittiioonnsraotfstdhoesperdesweintht stthuedtieesst,atrhteiclfeluaonrdocihnemalilcFa1l PgeFnOeSratwiaosn orabtssefrrvoemd FinOtdhoesseerda. rats. 30 Environmental Laboratory. SM Ensironmental Laboratory Page 15 Page 15 3M Medical Deparment Study: T-6295.25 Modal Doparmas Sty 29525 418-018:PAGE 1-389. Analytical Repor. TFRACNTE-0TO1X-0118609 Ava Repar FEACTrToxe169 Appendix A: Control Matrices and Test Article Table'. Characerizaton of he GonirlMatrices Usedfor Sera Anaysas in Study FACT TOX-169 [eTmO {Som comet |SomaGrom | Soman |SomaGrom [Ewmi|onovooveoaron| oworaoe | _owevaoii | ovovaoto [crom| caFtrommoo2m0roC| s| Frmomomo2n0C || _Frroman2n00 | Frozen-20C [o Fv e | fr os | ors so Table 5CharacterizationoftheTestArce Study FACTTOX-160 [mm|For rwoe wsre | [Sms| Cop Tosiyon ieDr Fon 510 riwso a aw EnmtonmantaLaboratory $1 Ewironmontal Laborators pages Page 16 3M Medical Department Study: T-6295.25 M4 Metal Deparment Stuy: T-6295.25 `Appendix B: Protocol and Deviations 418-018:PAGE 1-390 Analytical Report: FACT-TOX-169 LRN-EOI-0150 Anayical Report: F(ARCNT0T1O.X0-118609 au Environmental Laboratery $M Environmental Laboratory Page 17 Page 17 YE Lf 418-018:PAGE 1-391 marscr EO Demof bisrnedoh i Connetmeyn51t00S0) PHASE PROTOCOL. ME WeErEoSmiTnaEgiLnabsyorastoyriyes Sis St. Paul, MN 55106 eA pttsnhiant--ra--et M E o Su R y s25 LaboPrr ojea ctIt deno tifir caty ion 418-018:PAGE 1-392 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 Protocol FACT-TOX-L1R6N9-E01-0180 Study Identification DetermiMneavtailoonnoifctAhceidP/rCehsoelnecsetsernodlCCohnaclelnetnrgaetiaonnodfNPOeErfLluIonrvoeoscttiagnaetisounlfSotnuadtye i(nPFRaOtSs) in the Sponsor 33MM MCoerdpiocraalteDeTpoaxritcmoelnotgy BStu.iPladuiln,g M22N0-525E1-4042 Sponsor Representative Marvin. Case D.V.M., PhD. 33MM CMoerdpiocraaltDeeTpoaxritcmoelnotgy BStu. ilPdauiln,g M22N0-525-10424 Telephone: (651) 733-5180 Study Director Phase Locations P(rPian)cipal Analytical Investigator D3eManCnoarpJo.rLautcebTkoexricMoSl.ogy 3BuMilMdeidnigc2a2l0D-e2pEa-r0t2ment STte.lPeapuhlo,neM: N(65551)147437-1374 Kristen J. Hansen, Ph.D. Analytical Testing Laboratories 3M Environmental Laboratory B9u3i5ldBiunsgh2A-v3e8n-0u9e StPaul, MNS5106 Proposed Phase Timetable Analyses Start Date Analyses Termination Date P1a7c0e0 AEnlalmytSitrceaeltSSeEr,viScueist,eIn1c0.0 Minneapolis, MN $3614 January 20,2001 February 28, 2001 3M Environmental Laboratory $01 Environmental Laboratory Pogo2ors Page 19 meerHERO 1. Stupy 2 DPuevomsie E n fe Fer n d CocrioNnoStEPmeernssyie (FOS) ns `The analytical phaseofthis study is designed to evaluate levels of PFOS in sera specimens of sas ebony Ses FEoR Argus In-lifeStudy 418-018. All sera sampleswillbe 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. Te sP y oop n cre REGULATORY COMPLIANCE hSs UrSod Sa FdSDrnAdin CFR Part 58. +. Curves `The 3M Environmental LaboratoryQualityAssurance Unit will audit the study conduct, raw &. E Rese Sion e Fon FACTTOK168 coo Lamas Poi ep this protocol, and 3M Environmental Laboratory Standard Operating Procedures. Potassium Perfluorooctanesulfonate (KPFOS), CsF11SO3K, CAS 2795-39-3 TePsorpsevsodreneEsstileoklwr gooeny milienPoGmtgti)Svoey,eVfino six F1 pooled littersonday21ofgestation (at Caesarean sectioning), and from six F1 pooled f pes t fin seck - litters from day 5of lactation (after natural delivery). There are 312 sera specimens sent to Ems rmsas 418-018:PAGE 1394 3M Medical Deparment Study: T-6295.25 Analytical Report: FACT-TOX-169 Protocol FACT-TOLXR-N1-6E901-0180 Number of Specimens por Dosage group and Dosage Levels SonvdgLeovGeor n | a FCooRnsa, rsoann|| CFasIaRrroaPnanRSSecTtiNon| rormaron Lactation Bays | | FLramcetraetrisounpsD!aryo5m oemeeema 6 6T&5 T lea=s T eT ] = T&1] Pluss 1 [oT 1 [ l hl oloo na [ m =g T e1 r e T s T+1] To 1+ 71 [loa 1 o | [| F[aempserses[5 5|e [| [[oolfotsommpprrggrprrooss[ T 5 T6T 6& T 1 [[irllawmeemrpogrprrooss T 6T T e 1e &e 6] ] Col momorgrros 616 | 6 |6] 7. SPECIMEN RECEIPT lSiefreapshpaesceiomnenJsanwuialrlyb2e0r0e1ce(ievsetdimfartoedm)A.rgSeursaRsepseecairmcehnLsabwoerraetosrhiiepsp,eIdncb.yaotvtehreniegnhdot fcotuhreieirn and were received frozen on dry ice. Specimens were identified with sample numbers (lab rreegqiusetsetrnedumwbiethrsthoer3nMumEbnevrisroasnsmiegnnteadlbLyabAorrgautsorRyesaenadrtcrhanLsafbeorrraetdortioeas,frIenecz.e)rufpoornstreocreaigpet.,All tshpeecSiammepnlsesTernatctkoitnhgeS3yMstEenmviSrtoanndmaerndtaOlpeLraabtoirnagtoPrryocweedruerer.ecDeeitvaeidlaonfdspteraccikmeednacicnosrpdeictnigotno fporresdeanmtaegdei,n rtehceeipphta,ssetorreapogret,.identification, chainof custody protocols,and data will be 8. PREPARATORYMETHODS `ETCSo-m8p-o4u,nd"sExftrroamctSioenroufmPfootraasnsailuysmiPseursfilnugorHoPoLcCt-anEelseucltfroonsatperaoyr/oMbaessr SFlpueocrtorcohmeemtircya"ls Arneaalgyetnitciaslasdadmepdletos aarleaebxotrraatcotreydsuasmipnlgeaanndiotnh-epaainailnygteexftornacptaiiornipsrpoacretdiutrioen.eAd ninitoonM-pBaEir.inTghe 'MIBE extract is then removed and put onto a nitrogen evaporator uai dry. Each extracted laboratory sample is reconstituted in 1.0 mLof methanol and filtered through a 3 cm' plastic: syringe attached t0.2 02 um nylon fle ito glass autovials. 3M Environmontal Laboratory 3M Environmental Laboratory Page dof9 Page 2! 418-018:PAGE 1395 3M Medical Department Study: T-6205.25 Analytical Report: FACT-TOX-169 Protocol FACT-TOLXR-N16-9E01-0180 9. ANaLyTICaL METHODS ETS-8.5, "Analysis ofPotassium Perfluorooctaneslfonate or other Fluorochemicals in Serum Extracts using HPLC-Electrospray/Mass Spectrometry" oAnnalcyhsairsacitserpiesrtficoromafedpbayrtmiocnuliatrolriunogroocnheemoricmaolruespirngodHuPctLoCn-sESsMelSeMstSe.d fFroormeaxasimnpgllee,primary `molecular ion 499, elected asthe primary ion for PFOS(CF11SO5) analysis, s fragmented further to produce ion 99 (FSOx-). The characteristic ion 99 is monitored for quantitative analysis. Minor modifications o the preparatory and analytical methods may be implemented at the discretionofthe PAI. 10. DATA QuALITYOBJECTIVES 10.1 LTihneecaoreiftfiycientofdetermination (7)ofthesandard curve mst be equal 0 or grater than 0.990 using 1/x weighting. 10.2 L`TihmeiLtsOoQfwiQlulabnetietqautaliotno t(hLeOlGo)west acceptable standard inthe calibration curve, wdeiftihniend=a3s 0th%eolfotwheestesxtpaencdtaerddctohncteinstrbaottihon2.times the matrix blank and is calculated 103 Acceptable Precision `Quality control samples are required to meet + 25% precision, provided they are `quantitated within the slected calibration range. 10.4 UfsaeocofnfCiornmaftiorrmyamteotrhyodMeituhsoedd,sanamendmen1t0 this pretocol will be writen. 105 DPeFmOoSnisdterntaitfiicoatnioonfwSiplelcbiefiscubisttya,nteitact.ed by chromatographic retention time. (approximately 7.3 minutes), by the characteristic primary ion (499) and the characteristic product on (99). Minor modifications to the Data Quality Objectives may be implemented atthe discretionofthe PAIL " 3MEnvironmental Laboratory 3M Environmental Laboratory Page sors Page 22 3M Medical Department Study: T-6295.25 418-018:PAGE 1-396 Analytical Report: FACT-TOX-169 LRN-E01-0180 ProtocolFACT-TOX169 11. SPUaBc-eCAOnaNlTytRiAcCalTESeDrAviNcAesL,YISnIcS(Tie If Facility) will be performing the extractionofserum sLapbeocriamteonrsyffoorraannaallyysseiss.. APlalceexAtnraalcytteidcsaalmSpelrevsicwiel,lIbnce.swhiilplpfeodltloowthaell3aMppElnivcaibrloenm3eMntal EAnnavliyrtoincmaelnItnacl. LSatbaonrdaatrodrOypeSrtaantdianrgdPrOopceerdautriensg. Procedures and the following Pace: PSS-Admin-01 PPSSSS.-0DPCS--0051 PSS.OPS-02 PPSSSS.-0OPPSS--0034 PPSSSS.-OOPPSS--0056 PPSSSS.-0OPPSS--0078 PPSSSS-.OOPPSS--1135 PSS.OPS-16 PPSSSS--OOPPSS3312 EPSTSS-9O4PS0-37 PSS:MTR-01 PPSSSS:-MMTTRR..0026 PPSSSS-MMCI-TR0.108 PPSSSS-.ADRMC--0032 QLuaabliNtoyteSbyosotkesm, Logbooks, and Phone Logs Use and MaintenanceofUltra-Turax T25 Homogenizer HRaazwarDdaotuas Waste Disposal UUsseeoafndCoMmapirmeesnsaendcGeoasfeNs-iEnvtahpe LAanbaolryatitcoarlyEvaporator CUlseeanainndgMNaoinn-tdeinsapnocseabolfeNVAoNlOumpeutrriecIPiWpaettteersPurification System LUasbeoraantdoMryaiCnatlecnualantcieoonfsLab Refrigerators, Freczers, and Ovens UGlsaesasnwadrMeaCilnetaennianngce ofthe Fisher Scientific Isotemp Freezer UOpseeraantdionM,aiMnatienntaenocaencoefaSnhdakCearlsibrationofLaboratory OOppeerraattiioonn,aMnadinMtaeinnatnecnea,ncaenodfCaSlhiabkreartsiaonnodfVpoHrtMeexteMrisxearnsd(p3HMESlOePct)rodes CCaalliibbrraattiioonnooffCCeerrttiiffiieeddTWheeirgmhotmSeettersThermocouples VVeerriiffiiccaattiioonnooff CCaalliibbraattiioonnoafntdheUEspepoefnAdnoalryftiPicpaeltBtoarlances PurcofhLabaoratsoryiSupnpliges DSitastpiosstiictailonEvoaflAuracthioinveofMaDtaetar.ials 12. STATISTICAL ANALYSIS SEtxaatimsptilceaslomfetthheodcsalvciullatbieonlsimuisteedd itno tthhee acanlacluylsaetsiwoinlolfbmeeianncsluadneddsitnatnhdearpdhadseeviraetpioornts.. 13. RAerpepoorrttofthe analyte results willbe prepared by 3M Environmental Laboratory. The report vil include, but not be limited t0, he following, whe apolicable: 1133.29 NDaatmees uapnodnadwdhriecshsotfhethpehafsaeciwliatsy pineirtfioartemdiangndthceompphalseet.ed. 133 A statementofcompliance by the PAI addressing any exceptions to Good Laboratory Practice Standards. aM Envicnmental Laboratory 3M Environmental Laboratory Pago sols Page 23 418-018:PAGE 1397 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 Protocol FACT-TOXL-R1N6-901-0180 13.4 Objectives and procedures as stated inthe approved phase protocol, including any 13.5 aIdmeenntidtmy,enputrsittyo,tshaeboirliigyinaanldphtahseesoplruobtiolciotlyo.fthe reference standardunderthe 13.6 Aconddeistciroinpstoiofnoadfmtihneismteratthioodns. used 0 conduct the tst(). 1133.87 AA ddeessccrriippttiioonnoofftahneyscpiercciummesntsa.nces that may have affected the quality or the integrity 13.9 oTfhtehneadmateao.fthe PAI and the namesofother scientists, professionals, and supervisory 13.10 Aperdseosncrniepltiionnvoolftvehdeitnrtahnesfpohramsae.tions, calculations, or operations performed on the dcaotnec,luassiuonmsmadrraywannfdraonmaltyhseisanoaftlyhseesa.nalytical chemistry data, anda statement ofthe 1133..1112 TSthaetisstiigcnaeldmaentdhoddasteudserdetpooretvsoalfueaatcehothfetdhatea,inidfaipvpildiucaalbslcei.emists or other professionals 13.13 Tihnevlolovceadtiionnthwehpehraesrea,widfaaptpalaicnadblteh.e phase reportare tobe stored. 13.14 AisnstpaetcetimoennstapnredpaaurdeidtbsywetrheemQaudaleitayndAstshuerdaantceesoUfnaintylisftiinndginthges draetpeosrttehdattosttuhdeyStudy Director and Management. 13.15 Iafcicteipstende,cetshseacrhyatnogemsakviellcobreremcatidoensinorthae dfodrmioft1a0in.oafnminesanldrmeeponrttiastseureidtbhyasthbeeSetnudy Daimreencdtoerd.,Tthhee aremaesnodnsmefonrttvhiellamcleenardlmyeind,enatinfdywtihellpbaertsoifgtnehdebfiynatlhereSptourdtythDaitreicstboer.ing 14. LOCATIONOFRAW DATA, RECORDS, AND FINAL REPORT Ofarciigliintaatledaautdaitosrocfotphieesstthuedryeodfu,riwniglibtseparvoagirleasbsleanatd3beMfoErneviacrcoenpmteanntcaelofLatbhoerapthoarsyetroeport, b`Wehleonwtwhiellpbheasreetraeipnoerdtiins tchoempalrectheidv,esaolfl o3rMigiEnnavipraopnemrednattaal, iLnacblouradtionrgyt.hoAslelictoemmesslpiosntedding tprraoicneidnugrersec,oerdqsu,icpamleinbtraptrioocnerdeucroersd,s,anidnsmtertumheondtsmwaiilnltbeenarentcaeinleodgsi,nstthaendaarrcdhiovpeesraotfitnhge 3M Environmental Laboratory. 14.1 Tahcceorfdoilnlgowtiong3MraEwndvaitraonamnednrteaclorLdasbwoirlaltobreyreSttaainndeadridnOpteherasttiundgyPfroolcdeedruirnest.he archives 14.1.1 Approved protocol and amendments 1144..11..23 SShtiupdpyicnogmreescpoornddsence. 1144..11..45 RApapwrdoavtead final report (orginal signed copy) 14.2 14.1.6 Electronic copiesofdata The following supporting records willberetained separately from the study folder in the archives according to 3M Environmental Laboratory Stendard Operating. Procedures: aM Envionmantl Laboratory 331 Environmental Laboratory Pago 7of9 Page 24 418-018:PAGE 1-398 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 Protocol FACT-TOXL-R1N6-9E01-0180 1144..22..12 TCarlaiibnriantgironecroercdosrds 1144..22..34 ISntsatnrduamrednOtpmearianttinegnaPnrcoeceldougrses, Equipment Procedures, and Methods 15. DATA/ SAMPLE RETENTION `Amllaisnetraainsepdebcyi:mens remaining afte the analytical phase is completed will be sent to and D3aMveCoErhproerastmeanToxicology 3StM. PCaeunl,teMrNBld5g5124346-0C-148 TFealxe(p6h5o1n)e7(3675-14)775343-5070 16. PROTOCOL AMENDMENTSANDDEVIATIONS PSltaundnyeDdicrehcatnogreasntdotthhee pSrpootnoscoorl'swiRelpbreesiennttahteivfeo.rmAomfewnrdimtetenntsamwielnldbmeecaosnssiidgenreeddbays tphaert oifntdhiceatperdotionctohleapnhdasweilrlebpoerta.ttAanchyedotthoetrhcehfainngalesprwoitlolcoble.iAnltlhcehfaonrgmeosftwortihtetepnrodteovcioaltiwoinlsl,be signed by the Study Director and fled with the raw data. 17.ATTACHMENTS 17.1 AMattetraiaclhSmafeetnAyt:Data Sheets (MSDS) for Reference Standard 34 Environmental Laboratory $M Environmental Laboratory Page gof9 Page 25 3M Metical Deparment Sud: T-6295.25 15. SomaTuReES Yess, TGs `Marvin T. Case, D.V.M., Ph.D., Sponsor Representative 418-018:PAGE 1-399 Anaya! Report: FACT-TOX-169 ProcolFACTOKBRYED1-0150 3 porn 2tart Date MDeaanmnamZacaokelrCb3, SatudytDivetstors ziDlaeol CRl eineHare-- an, PAD. Proce Analyte Tvesigaor T 1/24/e 01 [---- Boy Page ooro Page 26 418-018:PAGE 1400 3MMedicalDepartmentStudy: T-6295.25 Analytical Report: FLARCNT--ET0OLX--0116S90 Record of Deviation T Tdantiication St"uFdyAC/TP-uTsjOiXt-N1o6:9,Lims #EOL-0IS0,Argus418.018 - - (DCehvieactkioonnTey)pe XSPrOotPocol (0MOetthheord: (JEquipment Procedure DoFdAuCiTnTenOXN-a1b6e9r~~ Daoeioloolfoceimencs TC Description: Re"TqhuepirroetdoPcroloscaetdeutrheal3p1r2oscpeecsisne:nswillb o eceved athe_ Environmeno tal Laboratory. A2c3t5usaplePcriomcesndsurweeiprreorceecsse:iveda the Enviromental Laboratory. (chaosamTi eActsinoendsd,TSaOkmePnr:eevisinon,ett) Thisdevisfonwaswriten. - sn ResoBry TT pg On A Was TV. Trpacton StuTPrdojeyct os/avln Noadver Tnpa onesHeomeof dy. Gh 0521/01 Authorized By Woes (SuFdy Daigeteor / roject Lead) ohe EBreTae oe Wp Tr 3g 00) DSaae rs pry 3M EnvironmentalLaboratory unt ns OctDevisateioknNoS. E_T__|RY SM Environmental Laboratory Page 27 418-018:PAGE 1-401 3M Medical Department Study: T-6295.25 Record of Deviation Analytical Report: FACT-TOX-169 LRN-EOL0IS0 1. Identification EFACT T-TOX-169,Lims#EOL-0180, Argus418.018 DeviationType (Checkone) osopr Wenod O'Equipment Procedure 0Protocol Other: DoEcTiSme8n4t2Number ~~ Dat20e1l501of-o2c20c/u0r1ence II." Description: Re1q)uSiercteidonP1r0o.c1e.d2aunrde1i0.1p.3.r2 oPcreses:pa_r_ie__w_at_erbanksandI tio maiBlanforkcasch stu| dy. 32))SSeeccttiioonn1111..11..42PDroepnaorteeaxtnrdascptliekseos nthea0sntan.darSdocfummrvaeLif.oreach study. | 4v)olSeucmteisoenqu1a1.l1o.2tThfemossatmpslaempvloelvuomleusm,esarelessthan 1.0 mL, extract standards with matrix |ActualProcedurerprocess: ee 21))FSoiuxrwaextterrabcltaendkcsuarnvdessiwxemreetphroedpabraedn-t kswerr weaidtdhe eidnidtaue leatnod hfeinallarvgoelnuummebseorfo0f.5smamLpalneds. | o3n)eMwaintyh sinaimtpalleasnwdefraelevxolutmerwsiaotfhcaLniOiWenf.dil volume less than 0 ml. Refer (6 the attached S4e)rFvoprlsmvoolsumseamlpolaeisg,h<t0s.o5nmstLfoorfsSeorbaelwvaoslauvmaielSablOe, MHAowe,ver, curveesewere nol prepared with sor teerl O e e] 1II. Actions Taken: Thisdevisonwoaos weite{n.u0 cehs amendment fssued: SOPrevision, eto) TT Recorded By Date Kelly Swartout 22101 IV. Impact on Study7 Project 31)&B2e)cauAsdedtihemtieotnhQaoCld iiss annotrschparroavcteemreiznetadnfdorwviolllinmoetas<d0v.5ermLs,aefsleamcyptlheissswitthuidnyal.volumes | l4e)sTshtihsanco0u.r5smeLofwialcltboen wlaasgtgaekdienntbheecadusaet{ah_e._eihodRasnot beer characterized or extractions<0.5mL. Robuofsthetcurnveeswisllnsot beadverselyaffected. retest ---------- -- bh oaiazor AuthorizedBy (Sidy Director/Projesi Lead) SHErrol Labret DoD viitlon Foa . Pom ETS 080 tlle (pettySy incrpry Lend Tr Tr) $3 EnsironmSehnatdayl DLarbeocrhart.orFyoeOca budber fimwerglobtrtin 2a 00 1 Page 25 418-018:PAGE 1-402 3M Medical DepartmentStudy: T-6295.25 Record of Deviation Analytical Repor: FLARCNT--ETOOIX--0116890 I. Identification SO Sudy PojeNo. Deviation Type (Checkone) Tagen = osop Bictiod O Equipment Procedure. OProtocol Other: Documen Naber ES B55 Dagefocumenes BT, TI." Description: or RequiredProcedwrefprocess:__________________ | Pring Fo M3_7a_talielautpuisorusd eink al| te aepel LooeA Gaie useCi p E A a A o fo. eooA J i Li ai e t rre E IK. i Optia E libkbimoddra'es| Df Loki Lasts Jon Esscakis Lovef r entai| cap Gecrre _ (suchas ame1nlld.meAncttiiossnused,TSaOkePnr:evision, etc) D`aecapdeps2aLesLieonrsts.eTaliSSeemhseicaellsergyraDttSeneFat, tor as, By |remeany 27% Ne advoas apgecte a TV. TriepadTtonStu7Prdojeyct pn oWlexiel ow A=n2y0)/ Rithorized By (StudyDirecor/ProjeciLead) Dae ro Teich Mo Coe Shy Slr Dror Uuabar Dlr opBoer 714001 kTGo EnvironmentalLabora ndJ ls sy re ees `Deviation NB. 3 $M Enironmental Laboratory Page 29 418-018:PAGE 1403 3M Medical Department Study: T-6295.25 3M Medical Deparment Study: T-6295.25 Analytical Report: FACT-TOX-169 LRN-EOL-0180 Analyical Report: F(AACNT-ET0O1X0-118609 Appendix C: Extraction and Analytical Methods "This appendix includes the folowing methods: EETlSe-c8t-r4o.s2p,raEyxtMraascstiSopneocftFoumoertoyc,he(m1i5caplagCeosm)pounds from Serum for Analysis Using HPLCESTeSr:u8m:5E.x2t,raAcntaslyUssiisngofHPPoLtCa-sEslieucmtPreorsfplruaoyroMoacstsanSepseucitfroonmaetteryo,r O(1t1hepragFeluso)rochemicals in $M Emironmental Laboratory Page 30 3M Medical Department Study: T-6295.25 : 3M ENVIRONMENTAL LABORATORY 418-018:PAGE 1-404 Analytical Report: FACT-TOX-169 LRN-E01-0180 METHOD EXTRACTIONUSOIFNFGLHUPOLRCO-CEHLEEMCITCRAOLSPCROAMYP/OMUANSDSS SFPREOCMTSREORMUEMTRFYOR ANALYSIS Method Number: ETS-8-4.2 Adoption Date: 03/01/99 Effctive Date: 32/2191 Approved By: pf Laboratory Manager lb to Group Leader 03afey Date ospoiser Date 1.0 Score AND APpLICATION 11 Scope: This method i for the extraction of luorochemical comoounds from serum. 12 Applicable compounds: Fluorochemical surfactants or other fliorinated compounds. TRY 13 MSatreicesm: Reabbni, ca, bovine, monkey, and human serum or otherfluidsas designated in EU 2O20 SummoFaMerTHOyD B21 Tshulifsopnaeterf(PoFrOmS)anourcseotd.hemreftlhuoodrodcehsecmriicbaelsstuhrfapcrtoacnetdsufrreofmoseerxuamc,toirnogtpheerrfiidosro,oiusainneg-an Ole & 8BIR ion pairing reagent tehve asanmspleanatnpdttheo and vaneal methyk-tert-butyl ether (MIBE). An ythe-tioonppaiErviasppoarrattiotriovnetd idnityo.MIEBaEc.h ion pairing reagent is added eTxhaecMtIsBeEseoxtmrascat eisd in to & 0 mL of methanol, then ered through a 0.2 um nylon ie using a 3-mL plastic 2Wort 695 Exrscion ofFsEcrrascshe4m2icas from Scum, Page ari $31 Environmental Laboratory Page 31 3M Medical Department Study: T-629525 : 418-018:PAGE 1-405 Analytical Report: FALCRTN--TEOOXL-0116590 sdyermionngsetrianttoedg:laPssFOaSut,ovPiaFlOs.SA(,ApPplFiOcSaAtoAn,oEf(tFhiOsSmEe-tOhHo,d tPoFsOeSvEeAn,flMv5or5o6c,heamnidcaalssuwrarsogate standard (see 3.0 Definitions). {22 Tmhetehsoed.sample extracts are analyzed following method ETS-8-5.2 or ther appropriate 30 DeeNTIONS 31 PROS: perfluorooctanesulfonate (anionofpotassium sal) C.F SO5 32 PFOSA: perfluorooctane sulfonylamide CsFirSO;NH; 33 PFOSAA: perfluorooctane sulfonylamido (ethyacetate CuFyrSON(CH;CH,)CH,COY 34 E(FOSE-OH: 2(N-ethylperfuorooctane sulfonamido)-cihyl alcohol CsF13SO:N(CH:CH;)CH;CH;OH 35 PFOSEA: perflorooctane sulfonyl exhylamide CifSON(CHyCHyH 36 MSS6: CiF,sSON(HI(CH:COOH) 37 Sumogate standard THPFOS: 1H-1H-2H-2H perfluorooctane sulfonic acid 40 WamnesawpCanons 41 Health and safety warnings 411 Uhasnedluinnigvaenrsiamlaplrteicsasuutei,onwsh,iceshpemcaiyalcloynltaabionraptaotrhyogceonast.s, goggles, and gloves when 50 INTERFERENCES 51 There are no interferences known at this time. 60 Eouruext 6.1 Thacecefpotalblloew.ingequipmentisused whileperforming this method. Equivalent equipment is 611 Vortex mixer, VWR, Vortex Genie 2 61.2 Centrifuge, Mistral 1000 or IEC 6.1.3 Shaker, Eberbach or VWR 61.4 Nitrogenevaporator, Organomation 6.1.5 Balance (0.100 g) 10 SussoMatemars 71 Gloves 72 Eppendorf or disposable pipetes, plastic or glass (or equivalent) 73 Polypropylene bottles, capabie of hokding 250 mL and I L (Nalgene or equivalent) $M Environmental Laborators Exuasion of Fl1sr5a8e4he2micl rom Sem Page 20015 Page 32 5M Medical Deparment Study: T-6295.25 418.018 PAGE 1-406 Analytical Report: FLARCNTC-ETOOLX--0116590 74 Volumetric flasks, glass, ype A 7.5 40 mi glass vias (CHEM or equivalen) | 7:6 Centrifuge wbes, polypropylene, 15 ml. F11708 LBaobtelles-top Dispenser~ 3.010 100mLor a 5-10mL graduated pipette, glass 79 Syringes, capableofmeasuring 5 uL 10 0 uL. 710 Graduated pipers 7.11 Syringes, disposable plastic, 3mL (B&D or equivalent) 72 Syringe fiers, nylon, 02 ym, 25 mm 713 Timer 7.14 Crimp cap gas autovials and caps 7.15 Crimpers Note: Prwaitoerr1.0 uRsiinnsgesgylraisnsgweasremnidnibmoutmso,fritnsieme3stiwmietshwmietthhamneotlh,an3orliansnesd3frtoimme3s sweiptahrate vials $0 REAGANDESTNANDTARSDS 81 Reagent grade water, Mill-QTM, Nanopure IT or equivalent 82 Sodium hydroxide (NOH), 1 Baker or equivalent 83 Tetrabutylammonium hydrogen slfate(TBA), Kodak or equivalent 84 Sodium carbonate (Na:COy), LT. Bakerorequivalent 85 Sodium bicarbonate (NaHCO), JT. Baker or equivalent 8.6 Methyl-T-Butyl Ether, Omnisolv, glass distilled or HPLC grade 88.87 MSeetrhuamnoolr,bOlmonodi,soflrvo,zegnlafssodmisstuilplpeldieorr HPLC grade 89 Fluorochemical standards 89.1 KPFOS (PFOS) (3M Specialy Chemical Division), malesular weight = 538 89.2 PFOSA (3M Specialty Chemical Divison), molecular weight = 499 $9.3 F5 FOSAAGH (PFOSAR) GM Spaciy Clic ivr). moll wiht = 8.9.4 EXFOSE.OH (3M Specialty Chercal Division), molculas weight = 570 89.5 PFOSEA (3M Speciaty Chemical Divison), molecular weight = 527 88.99.76 MSu5r5ro6ga(t3eMstSapnedcairadl:ty4C-hHe,mpiecrafluDoirvoioscitona)n,e msoullefocnuilcaracwiedi(g1h-tH=,15-5H7, 2-H, 2-H 8.9.8 COt4hFe1rSfOl,uHo,ro[cThHePrFiOcSa)l)as,, maoplpercouplrairatweeight = 428 $3 Emiromnental Laboratory Extactonof FrEsshseanrias rom Ser Pagesorts Pages 3M Medical Department Study: T-6295.25 418-018:PAGE 1-407 Analytical Report: FACT-TOX-169 LRN-E01-0180 810 Reagent preparation NOTE: Wphreepnarparteiopna,riandgjudsitfearcecnotrdvionglluym.es than listed in reagent, standard, or surrogate: . 810.111000N0msoLdibuemakheyrdrcooxnitdaien(inNgaO5H0)0:mWLeiwgathera,ppmrioxxiumntaitleallys2o0l0idgs NaraeOdHi.ssoPlovuerd.inSttoorae: ina 1 L polypropylene (or equivalent bottle 8.102 11 0NNsNodaiOuHm hsyodlruotxioindient(oNaaOH1)00: mDLiluvtoelu1m0etNriNcaflOasHk a1:n1d0.briMnegas10urveol1u0mme Lusoinfg water. Store ina 125mLpolypropylene (or equivalent) bot. 8.10.3o0f.5TMBAteitnrtoaabut1yLlavmomlounmieutmrihcycdornotgaeinnisnuglf5a0t0e (mTLBA)w:ateWre.iAgdhjuasptptrooxpiHmat1e0ulysi1n6g9 aapsptrhoexpimHatcehlayng4e4staob5ru4pmtlLy)ofDi1l0utNeN{0&OvHol.u(meAdwditthhewaltaetr.1.S0tmorLeonfaNa1OLH slowly, polypropylene (or equivalent) bottle. 8.10.3.1 TneBeAderdequusiirnegs |aNchNecakOpHrisoorltuotieoanch use to ensure pH = 10. Adjust as 8.10.4 0a.p2p5roMximsaotdeiluym2c6a.5rbgoonaftseo/dsioduimumcabribcoanrabtoena(tNe&b:ufCfOe)r (aNndai2C1.O0YNgoGfHCsOoyd)i:umWeigh bSitcoarrebionnaate1L(poNlGyHpCrOoyp)ylinetnoea(o1rLeqvuoilvuamleetnrti)cbfollasek.and bring to volume with water. 811 Standards preparation 8.111 Prepare PFOS standards for the standard curve 8.11.2 fPlrueopraorcehoetmhiecralflsutoarnodcahredmsiacraelasctcaenpdtaarbdlse, (afsoarpepxroapmrpilaet,e.onMeuwlotrikcionmgposnteanndtard PsoFlOutSiEonA,coMnt5a5in6inagndapEp(roFxOiSmEa-teOlHy).1.00 ugo/fPmFOlS, PFOSA, PFOSAA, 8.11.3 Wtheeiagchtuaaplpwreoixgihmta.teFloyr1s0ta0nmdgarodfs wPiFtOhSK-inotrooat1h0r0smaLi.s,vmoullutmieptlryibcyfalasckoarrnedctrieocnord fwaecitgohr.toFforPeFxOaSmp=le4,9t.heCmaollceuclualtearthweeicgohrtreocftiKonPFacOtSor=b5y38u,sianngdthtehefomlolloewciunlgar equation: mmoolleeccuullaarrw.i.KPPFFOOSS((459398)) 07 (Corectionrection acfaectror,) 8.11.4 Bring to volume with methanol for a stock standard of approximately 1000 pg/mL. 8.115 Daiplpurtoexitmheatsetloyck50soplgu/timoLn.with methanol for a working standard 1 solution of (10F00 matxt)) pm 8.11.6 Daiplpurtoex.wo5r.k0iungg/smtla.ndard | with methanol for a working standard 2 solution of 3M Environmental Laboratory Extracion ofFluEcToSsh8e4m2icas from Serum Page dos Page 34 418-018:PAGE 1-408 (mnt) nm 1 Sur EOS so tt pi (msmtont) approx. 0.50 pg/mL. gms CF13SO3H (THPFOS) into a50 mL volumetric flask and record the actual weight. Po1200 aps pg/mL. tsAeont gm ot Soma `and useasmethod blanks. ER Cont epmp hone 3M Medical Department Study: T-6295.25 418.018:PAGE 1409 Analytical Report: FACT-TOX-169 LRN-E01-0180 10.1:3.3 Scounrtrionguaitnegmcaatlriibxractiuornvvee--rIifficaatsiuornrosgaamtpelemsa:trix is used for the curve and k 4) prepare two (2) matrix blanks using the surrogate matrix. b) Amlastor,ixpsrpeipkaer/amearmiatxrsipxibkleadnukplwiictahtecaacthosneet loefvemla)tursiixnsgpiakceosm(maersceiailslay available matrixofthe same species a the study animals. 102 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: Mstautdryixansipmiaklesaanrdea nescuersrsoagrayteifmmaattrriixxiisunsoetd afvoaritahbelecuirnvteheansdamCeCsVpescaimepslaesst(hee.g. rtaobbmietassuerraabmlaeylebveeulsoefde0ndgoegneenroautes satnaanlydtaerdincutrhveemsofnokre2ymsoenrak)e.yPsreerpaasrteudayn,ddue tahnealeyxzteramcattiroinxfsopritkaergaentdamnaatlyrtiexs.spike duplicate samples to verify te accuracy of 10.2.4 Pcornecpeanrteraetaicohn)MuSsianngdasMaSmDpalte cthwoose(n2)blyevtehlesa(nuasluyasltl,ytaypliocawllaynsdemriadf-rarnagecoontmrol `group animal received with each sample set 10.2.4.1 Imfatthreirxespisikneostussuiffnigciceonmtmseerrcaiaalvlaiylabvlaeiflrbloemmtahtercioxnftrroolmgtrhoeups,amperesppaerceies as the study animal 10.2.5 pPerrep2a0resaomnpelemsa,trwiixthspaikmeiannidmumamtorif2x smpaitkreidxupslpiickaetsepart elaecvehllpeevrelbalticsht.ed iInf1a0b.2a.c4h linocwl,udoershmigohreratnhgaenl2e0velssa.mples, additional spikes may be arepared at the same, 10.2.6 1atfmaoprperoxtihmanat2el5y%2o-f5tthiemseasmtphleesexapreect<eLdOLQO,Qt.wo matrix spikes should be prepared 10.2.7 1afddtihteiomnaajlormiattyroifx tshpeiskaesmpslhoeusladrebea proerpaarbeodvetothaepphrigohxirmaantgeeosfamtphleeccuornvcee,ntrators. 10.3 Continuing calibration verifications (CCVs) 10.3.1 vPerreipfaircaeticoonntdiunruiinnggacnaalliybsriast.iPonrevpearrifeiacantdioannsaalmyzpeleCsCfoVr ifnsrtreuvmeernytasstsaabyilrituyn, regardless of the matrix type used to prepare the standard curve 10.3.2 Prepareeach continuing prepare the initial curve calibration (ic. either verification rom the study control tmhaetrsiax,mecommamterrixciuasledmattorix of same species, or surrogate matrix) $31 Environmental Laboratory ExuacionofFoEreTcShe8m4i2cals from Serum Page 015 Page 36 3M Medical Department Study: T-6295.25 418-018:PAGE 1-410 Analytical Report: FACT-TOX-169 LRN-EO01-0180 10.3.3 The expected concentrations of the CCV willfallwithin the range of the initial calibration curve; typically, the low to mid-rangeof the curve, + 10.3.4 cPornecpeanrter,aattioan,mipneirmgurmo,uptwoof 1c0onstaimnpuliensg.caFloibrraetxiaomnpvleer,ifificaatsiaonmsp,leeasceht a=t a34d,ieffiegrhetnt (8) checks are prepared; four (4) at a low range and four (4) at a mid-range concentration. 103.5 Additional continuing calibration verificationsmaybe included at the same, low, mid or high-range concentration. 11.0 CALIBRATION AND STANDARDIZATION 1.1 Prepare matrix calibration standards 1L11 Tarvaainlsafbelre |semraLiosfuaspepdr,opprreipaatreestewroumma0triax1b5lmanLksc,enutsirnigfutgheetvboel.umIefocofmsmeerracially available for most study animals. 11.1.1.1 Daveapieanbdlie,ngauspurornogsattuedymagtorailxs moraiyfabnealuysteed-ffroeregceonmermaetricoinaolfsetrhae cisalniobtration sattansdtaurddi,esFiofrqueaxnatmiptlaet,iornaobbfietxsterreammelayylboewulseevdelas<ofatshuerraongaaltyetemaitrriexquifroerd. 11.1.2 Tvfomlousmtessaeqmupalletvootlhuemessamaprleelevsosltuhmaens.1.D0 omLn,otexetxrtarcatctstlaesnsdatrhdasn w0i.t5h0mmiatroifx matrix. Record cach sample volume on the extraction sheet. 11.1.3 sWehrialebeptrweepearnianlgiqauottotsa.lof thirteen aliquots in 15-mL centrifuge tubes, mix or shake: 11.1.4 TTywpoica1l-lymLusaelitqhueotsst,anodraortdhecronacpepnrtorpartiiaotnesavnodlusmpei,kisnegrvaemaosucnutrsvelismtaetdriinxTbalbalneks| at the endofthis section to spike one standard curve,for a total of nine standards, two matrix blanks, and two method blanks. 11.15 RraenfgeerstoanvdaltihdeatLiionneraerpCoarltibErTaSt-i8on-4R.a0ng&eE(TLSC-R8)-5f.o0r-cVa-l1ib,rawthiiocnhculrivstess.the working 11.16 SSteaendSaercdtsi.on 13.0 to calculate actual concentrationsofPFOS in calibration 11.2 Tsuorrcoagcahtestwaonrdkaridn,gbsltaannkd,acrodnttoinauincg hcheiacckoe,nsavtnadenstacmopnlceenatdrdatainonap-phraotpfrailaltsewiatmhoiunnttheof ccaolnisbtraanttiocnocnucrenvterraatnigoenoifn2a.ll5sanmgp/lmeLs~a1t0a0p0prnogx/immLa.teUlsyua5l0l0y,nsga/mmLp.leosr aortehesrpickoendcfeonrtraation as determined by the analyst. 113 Etoxtersatcatblsipsihkeeadcmhaitnrtiixalsctaunrdvaerdosnftohlelmowaisnsgs1p2e.c6t-r1o2m.e1t6eor.fthis method. Use these standards 30 Environmental Laboratory `Exssion ofEFuTceoSc8he4mi2cals rom Sersm, Page 70615 Page 37 3M Medical Department Study: T-6295.25 418-018:PAGE 1-411 Analytical Report: FACT-TOX-169 LRN-EOI-0150 Table 1 Approximate spiking amounts for standards and spikes t Using1.0mLof matrix (approx. conc.) Hw analyte in matrix -- - T--1 Blank [o0S0| 0p1p 0 mT 0.0001005ppepmm [soo| pp 5m1 00pm [s00|pp 10 | m 0050pem [s S0 o0o |pp 5 p iom m1 | EE 0025500pmm 0.750 pr 120 ProcepuRe eee 12.1 Obiain frozen samples and allow (0 thaw at room temperature or in lukewarm water. 12.2 Laanableylst2i1ni5timalLs, pSoeleypartotpayclheendewcoernktsrihfeuegte(tAutbteawcihmhenttheAstourdysinmiulmabrewro,rkssahmepelte)IfDo,rdate and documenting the remaining steps. 123 Vortex mix for 15 seconds, then transfer 1.0mLor othr appropriate volume to the 15 mL polypropylene centrifuge tube. 12.4 Rerum unused samples to freer after extraction amounts have been removed. 12.5 Rwoerckosrhdeetth,e initial volume on the extraction worksheet (Attachment A or similar 12.6 SStpainkdeaarldl assamdpelsecsr,ibbeldanikns1a1.n2d.standards, that are ready for extraction with surrogate. 127 dSepsickreiabecdhinca1l1i.b1r,atoironTasbtalneda1irdnmtahtaricxtwiiotnh,thfeorepthperocpalriibtreataiomnoucnurtvoefssttanadnadradrsd. asAlso prepare matrix spikesif necessary and continuing calibration standards. 128 Vsaomrptleexsmfioxr h15essetcaonnddasr.d curve samples, matrix spike samples, ard continuing calibration 129 Check to ensure the 0.5 M TBA reagentisat pH 10. If no, adjust accordingly. 1210 To each sample, add | mL 0.5 M TBA and 2mLof 0.25M sodium carbonate/sodium bicarbonate buffer. 12.11 bUustiynlgeatnhebrottle top dispenser or 5-10mLgraduated glass pipette, add 5 ml. meshy-tet12.12 Cap each sample and put on the shaker at a seting of 300 rpm for 20 minutes $3 Environmental Laboratory Exacion of FElrursasch4em2icls rom Serum Pagers Page 38 3M Medical Department Study: T-6295.25 418-018:PAGE 1-412 Analytical Report: FACT-TOX-169 LRN-E01-0150 12.13 Centrifuge for 20 to 25 minutes at a seting of 3500 rpm or until layers are wel separated. 12.14 Labelafresh 15mLcentrifuge tbe with the same information as in 12.5. 12.15 Remove 4.0mLoftheorganic (top) layer to this clean 15mL centrifuge tube. 12.16 Putcach sample on the analytical nitrogen evaporator unl 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 lonifgneeederd. 12.19 Lnaubmeblera,1a.n5mimLalglnausmsbearutaovnidalge(nodrerl,ows-avmoplluemteimaeuptooivnita,l mwahternixn,efciensalsasroyl)vewnit,thextthreacsttiuodny date, and analys(s) performing the extraction. 12.20 SAtuetpac1h2.21285tommth,is0s.y2riungme.nyFliolnlemreisnthoftilhlerlatboeale3dmauitosvyirali.nge and transfer the sample from 12.21 Cap and store extracts at room temperature or at approximat4elCy unl analysis. 12.22 Cthoempstluedtyebtihnedeerx,traction worksheet (Attachment A or similar worksheet) and include in 13.0 DATA ANALYSIS AND CALCULATIONS 13.1 Calculations 13.1.1 CcaallicburlaattieonacsttuaanldacrodnsceunstirnagtitohnesfoofllPoFwOinSg,eoqruaottihoenr:applicable florochemical, in Lfsidb+ofil osfuxucrornocgeantriadt+ionfniaslpemlairinxvolkume)mL). Fingl! concenrason(ks /mi)eoff PPRROOSS in marix 14.0 METHOD PERFORMANCE. 14.1 TfohrespmeectifhiocdMdDetLectainodnliimmiito(fMqDuaLn)tiitsatainoanly(teLOaQnd)mvaatlruiexs s(pseeceifAitc.taRcehfmeernttosMBDaLndreCp)o.rtAt the discretionof the PAL, MDL may not be defined for some studies 142 Tthheeqfuoalliltoywionfgtqhueaelxityraccotnitornolansdamapnalleyssias.re extracted with each batch of samples to evaluste. 14.2.1 Method blanks and matrix blanks. 14.22IdfuspluircoagtaetseammpaltersixtoisvuesriefdyfeoxrtrcaucrtvieonanefdfiQcCi,encmya.trix spike and matrix spike 14.2.3 Cinointitailncuailnigbrcaatliiobnrcautirovne.check samples to determine the continued accuracy ofthe 143 Refer to section 14ofETS-8-5.2 for method performance criteria 5M Environmental Laboratory ExtractionofFusErTocSh8em4i2cals rom Serum Pagegorts Page 39 3M Medical Department Study: T-6295.25 418-018:PAGE 1-413 Analytical Report: FACT-TOX-169 LRN-E01-01580 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Dispose of sample waste, flammable solvent waste and used glass pipette waste using ` proper methods and by placing each in their designated waste container. 16.0 RECORDS 16.1 Complete the extraction worksheet attached to this method (or a similar worksheet), and. include in the study binder. 17.0 ATTACHMENTS 17.1 Auachment A, Extraction worksheet (a similar worksheet may be substituted for Attachment A) 172 Attachment B, MDL/LOQ values and summary 17.3 Attachment C, Ton Pair Standard Curves worksheet (a similar worksheet may be substituted for Attachment C) 18.0 REFERENCES 18.1 The validation report associated with this methodisETS-8-4.0 & 5.0-V-1. 182 FACT-M-3.1, "Analysisof Serum or Other Fluid Extracts for Fluorochemicals using 'HPLC-Electrospray Mass Spectrometry" 183 ETS-8-5.2, "AnalysisofPotassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" 9.0 AFTECTED DOCUMENTS 19.1 ETS-8-5.2, "Analysisof Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" 200 REVISIONS Revision Number 1 Sesion 1221 Changed includeResaampsloenstForoagreRatervoiosmitoenmperate. `Secon 12.13 Adthdeshaekerpdeed. `Seton 12.17 Final volume is 1.0 mL ot sesedfo initial volumes les haa 1.0 ml. 2 Secion L1,2.1 Eininaie poussiun' from perlucroocanesulfonate 89.1 AUK 10PFOS. 89.3 Add H1o PROSAA 102.1024, 142.2 Addiformasion abou sin suvogte mati,clarifycarol maieix and spiking he cure, change spikes to cvry20samples 10.3 Clarify purposeof coining albanchecks 11.1 Added wordingabout conol animal srs. using ed ot ingcommereally valble Revision oDuante 1 general, make les spfeorecquiipmefndsiuppcis. Subsite waterfo MilQl Nalgene or equivalent bolemaybe used. 5M Environmental Laboratory ETS:8.42 Exiracionof Fluorochemicais from Serum Page 100f15. Page 40 418-0I8:PAGE 1414 3M Medical Departmen Study: T-6295.25 Analytical Report: FACT-TOX-169 Extraction Worksheet ETS-8-4.2 Vivi tm SMuaiuir.. sScwuralopg/ems.d|| apFprCoxM. n05| | aFppCrenM.5|| apFproxC. 50 | Gomme| Veonyted Bwoxs. . pre f blee pre DaAtnaStpiked r wyr r r r r wr o] TTrr r rrr r Tr r T rrr F r Tr r Tr---- r ----1 r rrr r -- r r r r r r rr r r r r r r r r rr T T rr TT -- -- r E r S PS RSA A || ee Ei E RE A ---- Ir r rr Hei si whse FO Foie | mo 03M TOA GHI0 gi= Fr mLoTi 02 NCO FM NaHCO ber DremeLloimoe te her Dime ETL Sav Si iy Sie et Fr m-- -- 1 -- -- Hd cer Volume Deemer "Con. Cal Vercautseosnmsemtx afor 4 ure AppendiAa: Extraction Worksheet $31 Environmental Laboratory ETS842 1 Page lors Page 41 3M Medical Department Study: T-6295.25 418-018:PAGE 1-415 Analytical Report: FACT-TOX-169 LRN-EOL-0180 MCDoLm/poLuOnQd|valuMesDfLor|rabbiLtOseQru|m Linear Calibration Range (LCR) ` (ng/mL)| (ng/mL)| tAhpeprSotxainmdaatred CcaolnicbernattriaotnioCnusrtvoebe used for preparing S ng/mL~1000 ng/mL S ng/ml~1000 ng/mL. [PFOS 3.4A 6|A2_05| [5 ng/ml-1000 ng [EFOS11E .4 -|O3H 62| [5 ng/mL1000 ng/ml. [Mss6 16|0193 2 [Sng/ml-1000ng [ 'MDLP ILOGF v|alue5O s 7in1atS .|boviE n1e,8mo2A nke|y,Sanndgoimmaln-s1e0ru0m,0anngd/mLmonkey pas 7awerneotSastcally dCuertveersmioneddetTerwmoinceuerqvueisvainlecnacceh.ofRtehsepsonesmeastriinchesewraet,ebeoxvtirnaec,temdonaknedya,naalnydzhedumiatnhwtehreeraebqbuiitvasleerntu(m0 the Fabbit responses, therefor, ther MDL and LOQ will be the same valuesa determined i rabbi serum. PleaseseeLOQ Summary and MDL study in ETS-84.0 & 5.0-V-1 for further information. AppendiBx: MDLILOQ Values andESxuwmamcaiornyofFuEroTsShe8m4i2cals rom Serum 3M Environmental Laboratory Page 120015 Page #2 418-018:PAGE 1-416 ---------- | = | TEE [[oivmeen|monn[|oowonm||wowo || swoomn| =----|=---- p------ | | == 3M Environmental Laboratory Te Page 43 418-018:PAGE 1-417 ; J oS=R |r TaT [| c--re| = | -- | IE p----r i | i m FE I. es 418-018:PAGE 1-418 fm, Analyst(s) EXAMPLE Ton Pair em Standard Curves - Fluids `Studynumber: JSEoonDra on BX EEn r FCmixstd approx. S00 PRIS 0 vor ano pps - heextn caren E a EtEtttE e en Tic E ae ] oL S [Tos00| os07 | 0532 0501 _|_0521 |_0s01 0020 T1025| [TsooW|--s3%27| sor_| sa soi| ool 1.015 [C500Bogso enSr prygBm su [0020 1.02|5 [soo HW sor 3 "sso Hf Si8@5oF ASOT| 000s 1.010 [sooBsa 7Th SolBSATRon RBNeed | 0010 1015 [500 | soi|saa |sor [sai [sor 00151020| fIhe p A HlI An Calculated concentrofasttaindoanrdss ARin the sera lnmatrix: eErERabbit WN (Tass TT s001sa4 T404Tsoi sipTio 1 ooos | [as grey Suro [orsHs 8 3 - ngmL. [aH 29 /% 2688 #6B Bod Qos,| so | Bos 766 | | tadtsseopp t cn | aEpL a at ie wo DZem Duroetom 3M Medical Department Study: T-6295.25 : 418-018:PAGE 1-419 Analytical Report: FACT-TOX-169 LRN-E01-0180 3M ENVIRONMENTAL LABORATORY : MerHoD ANALYSIS OF POTASSIUM PERFLUOROOCTANESULFONATE OR OTHER FLUOROCHEMICALSIN SERUM EXTRACTS USING HPLC-ELECTROSPRAY/MASS SPECTROMETRY Method Number: ETS-8-5.2 Author: Lisa Clemen, Kris Hansen Approved By: Adaption Date: 0301/99 Revision Date: 2) / [01 Laboratory Manager Due Le fe Group Leader oz/eife) Date TechnAicaal RevLieCwera ve 2Dactoel. ol R 5 o 8 11 uSscionpge:HPTLhiCscmleectihroodsdpersacyrmiabsess tshpeecatnraolmyestirsyo.f serum extracts for luorochemical surfactants 916 =2 12 AoptpclicoanibzlaebCieomcpoomupnodusn:dssorcherical surfactants rer fuornatd compounds, or BJ 3Q 13 Mvaaltirdiacteiosn:reRpaobrtb.i, rat, bovine, moniey. and human serum, or other idsas designated in the 325 Word 695 ETsss2 Page tort Anyi of Seam Exact Using ESAS 3M Environmental Laboratory Page 46 3M Medical Department Study: T-6295.25 418-018:PAGE 1-420 Analytical Report: FACT-TOX-169 LRN-E0I-0180 20 SummoFAMETRHOYD. 2.1 pAelrtfhoorumgahncseu-ppboarsteeddmbeythaodva.liCdaarteifounlfaotrtemnotsitoncsohmomuoldnlbye upsaiedd tmoatmreitchso,dthQisCisasathere is great : evaxrtiraabcitlietdyfirnosmersa.erTuhmisormetthheordfdleisdcsr,iubessintgheHPaLnaCl-yesliescotfrfolsuporraoyc/hmeamsiscaslpescurtfraocmteatnrtys, or cShiamrialcatresryissttiecmoafsaappaprrtoipcriualtaer.fluTohreocahneamliycsails,issupcehrfaosrtmheedpbeyrfmlounoirtooorcitnagncasuslifnognleatieon(PFOS) atnoifount,hmeirzv=er4i9f9y.thAeddiidetnitointaylloyf,ascamopmlpeosumnadybbyedeatneacltyiznegdduasuignhgtaertaionndseomfmtahsespsapreencttrioonm.eter 30 Depmvmions 3.1 quAadtrmuopsoplheesryisctePmrsesaslulroew Ifoornivzaartiioouns (meAtPhDo:dsThoef iMoincirzaotmiaosnsbyQuuattiltirzoinIg vaanrdioUulstsiomuarcterisp,le Aprtomboessp,haenrdicinPtreersfsacuerse.cThheemisceailncIlonuidzeatbiuotna(reAPno)t,liTmhietremdotso:prEalye,ctertocs.prTahyeIoinoniizzaattiioonn(pErSoDc,ess inthese techniques occurs at atmospheric pressure (ic., not under a vacuum). 32 presEsulreec,trwohseprreabyylioonniszaintisoonlu(tEiSon, aEeS)t:raansmfeetrheoddtoofthieongiazsatpihoanspeevrifaortimneydcahtaragtemodsdprhoeplreitcs. "These charged droplets are produced by the application of a strong electrical field. 33 (MSM/aMsSs):SpTehcterAoPmIetQruya,tMtraosIs SepnedcUtlrtoimmeatteirpl(eVSq)u,adTraupnodleemsyMstaesmssSapreecetqruoimpepteedrwith qrautaidor(ump/ozl)eamnadsssusbesleeqcuteinvetldyetdeectteocrtse.d.ToAnssianrgelseeMleSctimvaelyybdeisecmrpimlionyaetdedfbory fmoansdsetteoccthiaorngoer a series (MS/MS) for more specific fragmentation information. 3.4 quadCrounpvoelnetsiyosntaelmsvs(.poZs-tsp1r9a9y8)purobteiiantl"eZrifsapczrea:ey"Thceonlftoersmtatmioodne.lsTohfeMsipcrraoymeamsisttterdifprleom a pdirroebcetliysaotrtthheogcoonnaelaopertthuerce,onaetearpepratusrsei.ngItnhrtoheugchonavetnotritounoauls pcaotnfhowramyaitniotnhetciosuanitmeerd. e`lmeacitnrtoedne.ancTehaoruegthhesthaemceo.nfiHgouwreavtieorn,iZs-dsipffrearyenct,omtphoenmeentthsodasnodfcoonpveernattiioonn,alclceoamnpiongn,enatnsd are: ncootmpcaotmipbalteibwliethwistohmeonoethaenrotZh-esrp,rbauytsoynsltyewmsi,thetcs.i)milar systems (i... Z-spray components ase 35 QuatMtarsosILtyinplxeSqoufatdwraurpeo:leSsyysstteemms.sofCtuwrarreentdleysiMgansesdLfyonrxthheasspWeciinfdicoowpser9a5tiaonndoWfitnhedsoewsNT 4i.n0stvreurmseinonts.(MiAclrlovmearssisoQnsuaatrtersoimIilaorr.UlFtoirmmaotrriepldeetqauialdssreuepotlheeMmaasnsuLaylnsxpeocrifMicasosLtyhenx NT User's Guide). 40 WARNINGS AND CAUTIONS 41 Health and Safety Warnings: 411 Uemspelcoayustiaovnolwtiatgheothfe avoplptraogxeimcaatbelelsy5fo0r0t0heVoprloib.e. When engaged, the probe Word 695 3M Environmental Laboratory Ansys of SerEuTmSEx8t5ra2ct Using ESAS Page20f 11 Page 47 3M Medical Department Study: T-6295.25 ) 418-018:PAGE 1421 Analytical Report: FACT-TOX-169 LRN-EOI-0180 412 Wanhdecnlohtahinndgl,ing samples or solvents wear appropriate protective gloves, eyewear, 42 Cautions: 421 Dtohenotbaocpkerparteesssuorleveenxtcepeudmsp4s0a0bboavre, ctahpeaHciPtyLoCfw4i0ll0nbiaart(eS8a0u0topmsai)tibcacskhuptrdeoswsunr.e. 422 Do ot run solvent pumps to dryness. 5.0 INTERFERENCES. 5.1 TStoormaigneiomrizaenyinptaerrtfeorfenicnesstrwuhmeenntaantailoynztihnagtscaommpleesis, Tceofnltoanctshwoiutlhdtnhoetsbaempulseedorfoerxtsraamctp.le 60 Equipe 6.1 E`mqoudiipfimceanttiolnissitendthbeelaoww mdaaytabaesmomdeitfhioedddienvioartdieorns(0 optimize the system. Document any 6.1.1 MwiitchroamnaeslsecQturaotstprraoyIioonrizUalttiionmasoruprlcee quadrupole Mass Spectrometer equipped 6.1.2 HcoPm1p1a0rt0moerntA,gialnedntalutoowspaumlpsleersolvent pumping system, solvent degasser, column 7.0 SUPPLIES AND MATERIALS 7.1 Supplies 7.01 Hniitgrhogpeunristyysgtreamd)e nitrogen gas regulated to approximately 100 psi (House ar or 7.0.2 HinPtLheCraanwaldyattiac.al column, specifics tobedeterminedby the analyst and documented 7.13 Capped autovials and capped 15mLcentrifuge tubes 8.0 REAGENTS AND STANDARDS 81 Reagents 8.1.1 Methanol, HPLC grade or equivalent 8.12 Meqiulivla-lQenTMt,waatnedr,maalyl wbaetperrouvsieddeidnbtyhiasMmieltih-oQdTsOhoCulPdlubsesMyistle-mQoTMr owtahteerrvoerndor 8.13 Ammonium acetate, reagent grade or equivalent 82 Standards 8.21 pTyrpeipcaarleldydtuwroinmgetthheoedxtbrlaacntkiso,ntpwroocmeadturriex. blSaeneksE,TaSn-8d-e4i.g2h.teen matrix standards are 30 Environmental Laboratory Ansys of SerEuTmSEx8t5ra2ct Using ESMS Page sail Page 48 3M Medical Department Study: T-6295.25 418-018:PAGE 1422 Analytical Report: FACT-TOX-169 LRN-EO1-0180 2.0 SAMPLE HANDLING 9.1 Farreesshtomraetdriixn csatapnpdeadrdasutaorveiaplrseoparrceadpwpietdh 1e5amchLacnaelnytsirsi.fugEexttruabcetseudntsitlanandaalrydsiss.and samples 9:2 aIfpapnraolxyismiastweillyl 4be Cd,eloaryeadt,reoxotrmactteemdpesrtaatnudraer,dsunatinldasnaamlpysliessccaannbbeepreerffroirgemreadt.ed at 10.0 QuaLITY ConrrOL eee eee 10.1 Solvent Blanks, Method Blanks and Matrix Blanks 10.1.1 Soonlcveendturbilnagnktsh,e mceoutrhsoedofblatnhkessatnuddymtaotdiextbelramnikns iafrceopnrteapmairneadtiaonndoacncaulryrzeedddautrlienagst sample prep. 10.1.2 Analyze at least one solvent blank prior to cach calibration curve. 10.1.3 eMxattrraictxs.blanks should be analyzed with cach sample ls that includes undiluted 102 Matrix Spikes 102.1 sIfecrua,rvseasmpalnedsmaertehmoodnQkeCy asreerap),rempaatrreidxisnpaiskuersraongdatmeatmraitxrsipxik(ee.gd.upcluircvaetsesnarreabbit epxrterpaacrteidoninefbfliacnieknscya.mple matrix (.g., monkey sera) and analyzed to verify 10.2.2 mIfacturrivxesspiaknedsmaertehroedquQirCed.are prepared in the same matrix as samples, no additional 10.3 Continuing Calibration Verifications (CCVs) 10.3.1 tChoentcianliubirnagticoanlicburravtei.on verifications are analyzed to verify the continued accuracy of 10.3.2 Asanmapllyezse,twwiothcaalmibirnatiimounmstoafndtawrodspe(ronbeatactheaancdh aolf2waylesveflisn)isahfitnegraenveirnjyeoctnieonto ten sequence with at least two calibration standards, 11.0 CALIBRATION AND STANDARDIZATION. 11.1 Awinlallbyezepltohteteedxtbryacltiendeamrartegirxescsailoinb,rawteioinghstteandda1/rxd,soptriofrortcoeedatchhrsoeutgohfzeexrtor,acutss.ingTMhaescsuLrvyen.x or other suitable software. 11.2 Trfeatnhaelcyuzrevtehdeosetsanndoatrdmecuertver.equirements, perform routine maintenance, reextract samples, of 113 Fraonrgep,uirpomsaeys obfe anceccuesrsaacryywthoeunsequcaintthietrattihneglloewveelndoforantahleytheaigh tehnedolfimtihtseofcatlhiebrcautrivoen.curve: aaptphreorxitmhaantetlhy f1u0lpapnbgoefoafnatlhyetes,ttanmdaayrdbecurbveen.efiEcxiaalmptloeg:enwerhaetne &tctaelmipblriatnigontocquuravnetitate cpopnbsitsoti1n0go00ftphpeb).staTnhdiasrdwsilfrreodum5ceppibnatcocu1r0a0cypaptbtrriabtuhteerdtthoanlitnheearurlelgrreasnsgieonofwetiheghctuirnvgeo(f5 $M Eninonmental Laboratory Anaisof SEerTumSE8xt5ra2ct Using ESS Pagesorit Page 49 3M Medical Department Study: T-6295.25 418-018:PAGE 1-423 Analytical Report: FLARCNT--ETOO1X--0116890 haingdhacohnicgehncturravtei.onIfstthainsdairddso.ne, inoalmsooraecctehpatnabolnee(p0oibnrteaskhotuhledlibneeaursreadnigneciontmomaolnowbectuwreveen tphpebcaunrvdest.heFhoirghexcaumrpvlee,matyheilnoclwucdeurtvheempoarynsi:nc2l5u0d,et5h00,fo7l5l0o,wi1n0g60po,in1t2s5:01,pp5b,.25. 100, 250 120 ProcepuREs 121 Acquisition Set up. 12.11 dOensitghneatMoarsesvLery,maosta2n dpiagghets,osfetsutp ayesaarm-pmloe-dlasyt,naamned.a sSatvre tthheatiswitllasiincnrsetarsuement through the alphabet with each addtional lis or tat day. Example Sample List: IYYMMDDa orDO10712a IYz=iynsetarurmeonfttensatm(:01()D for "Davey") MDDM==dmaoyntofhtoefstte(s1t2)(07) athierestxsta"mplaendlssto(orunn))ofthe day (the nex sample lst will end with `0. 12.1.2 Adasys,igannda fai3l-edniagmte suesqiunegnttihaelifnlsetrnuummenbterdetshiagtnsattaorrtstweirth, t| haendlasitn2credaigsietssboyfoynecarf-omrocach filename. Example File Name: [YYMMDD### or DO10712001 IYsi=nysteraurmoenfttensatm(e01()D for "Davey") MDDM==dmaoynoifheosftt(e1st2)(07) #t=3.digit sequential fle number starting with | through 999 (001) Aboltstol,eansupmabretro,fatnheinsjaecmtpiloen svho,luamsesaignndasmaemtphloedde(sMcSri)ptfioorns.cquiring, an net fl, 12.13 sTeoleecrtcSatIeRa(mSeintghloedI,onclRicekcoorndiMnegt)hoordMEdRitMor(bMuuttloinpien RtehaecMtiSonSMtoantiutsoPriannge).andSet sTeotnitzheatiaocnquMisoidtieonasstaaprptaronpdrisattoepatnidmems.asSsavt0e4a9cq9uiosriotitohnemeatpphroodp.ri1ateMSmTasMsSe.s. Also cionlsltercutmeedn.tsSeareeMeimcprloomyaeds,s aMdadsistiLoynnalxpGrUodIuDctEjToOn fDraAgmTeAntAatCiQonUIinSfIoTrmIaOtiNonfomray be additonal information and MRM. 12.1.4 Texytpriaccatleldy mthaetrainxalsyttaincdalarbdast.ch run sequence begins with solvent blanks and a set of 12.15 SSoalmvpelnet ebxltaenkascsshaorueladnableyazneadlwyiztehd tpewroioCdiCcaVllyintfoecmtoendietvoerr2yososniebl(e0antaelnystaemcpalersr.yover and are not considered sample exacts but maybe includedas such. $31 Environmental Laboratory AnyiofSerEuTmsEsxse2s Using ESIMS Pugsser Page 50 3M Medical Department Study: T-6295.25 418-018:PAGE 1424. Analytical Report: FACT-TOX-169 LRN-EOL-0180 122 Using the HPLC 12.2.1 Set up sample tray according to the sample lst prepared in Section 12.1.1 : 12.2.2 Sapeptr-oupprtihaeteHfPorLoCpttiomtahlerfeoslploonwsien.gRceocnodirtdioanctsuoarl caoncdointdiiotnisonisn tthhee ainnasltyrsutmecnotnsiders logbook: 12221 Samplesize = 10uL. injecton 12.2.22 Inectsampl=e | 12.2.2.3 Cycle time = 10.0 minutes 12.2.2.4 Flow rate = 300 pLimin 12.2.2.5 Mobile Phase (program) 2.0acmetMateA(minmo0n)ium [000min 10% [00%| [L00min[710% 100%| [SSomin [ ose [5%| [70min [ose [5%| [B00min | 10%[00%| 12.3 Instrument Set-up 1231 Refer to ETS-9-24 formoredetails 12.3.2 Check the solvent level in reservoirs and refill f necessary. 12.3.3 tChheectikp.thTehsetatiinplseshsousledelbecafpaitllwariytha ntohjeaegngdedofedtgheesp.rIofbet.heUtsiepasnfoeyuenpdietcoebet:o check upnrsoabteissfhaocutolrdy,bedicshaescskeemdblweeetkhleyp.robe and replace the stainless steel capillary. The 12.3.4 SOebtseHrPvLeCdrpopluetmstpcoo"mOinn"g,oSuettofthethfeltoiwp toof t1h0e-p5r0ob0e.L/minoras appropriate. 12.3.5 Taruorunnodtnhtehetnpitorftogheen.prAobfei.neRmeisatdsjhoutushletdbteipeoxftpehlelepdrwoibtehinfononimtirsotgeisnolbesaekrivnegd. 12.3.6 Tchhaenignestirnuomrednetru(s0eosptthiemsiezepatrheamreestpeonarstse:the following seitings. These setings may 12.3.6.1 Drying gas 250-400 ltershour 12.3.6.2 ESI nebulizing gas 10-15 ltershour 12.3.6.3 HPLC constant flow mode, low rate 10 -500ul/min 12.3.6.4 PHrPeLssCuire <op4e0r0atbianrg(cTohrriesctplya.r)ameter is not st, it is a guide to ensure the. 12.3.7 Cfaurrtehfeur.llyCognuniedcetthteheprvoolbteagientcoatbhleesop1e0ntihneg,proIbnes.et probe uatl i will not go any 331 Environmental Laboratory Ansys of SerEuTmSExetsra2ct Using ESIMS Page a 11 Page 51 3M Medical Department Study: T-6295.25 418-018:PAGE 1-425 Analytical Report: FACT-TOX-169 LRN-EOI-0130 12.3.8 soPgru.imnmtatrheytpuangeepnagde,sMtoSr fiinle,thHePsLtuCdypabrianmdeetrewrist,hs2amcpolpeyltisatpaenddintthoe tMhiecrnotsrouftmeWnotrd 1239 Clik on start bution i the MassLynx min page (his may vary among MassLynx nveurmsbioenrs,insceleudaepsprlolprsiaamtpelMesastsoLbyenaxnaUlySzEeRd.'S GUIDE). Ensure start and end sample: 130 DATA ANALYSIS AND CALCULATIONS 131 Calculations: 13.11 Calculate matrix spike percent recoveries using th following equation Recovery = QObuseermved RReeh-sMautrrilslBaontkk bReess 1 13.02 Caleulte percent iffeence using the following equation %Difoerrenmce == Eee E Cone Ci aleited Cone. 00 13.14 Calelate actual concentation of PFOS, or other lorochemical, in matrix (ug/mLy (CoinnciotfalPVRoOlSumCelofe fMroomrSeidu.ls)Curv+oe] ng/ml)Serxo_DrieluSiaonndoFra)ir) 11 100075 FiatVoime (nL) 140 14.1 METHOD PERFORMANCE Method Detection Limit (MDL) and Limitof Quantitatione(LOQe) aceemethod,sanaleytem, aned maurix specific. Please sce ETS-8-4.2, Attachment B, fora ising of current vlided `MDL and LOQ values. 142 Solvent Blanks, Method Blanks, and Matrix Blanks 14.2.1 aScotlivveentstbalnadnakrsd,imetthheocdalbilbarnaktsi,onancudrvmea.trix blanks valles must be below the lowest 143 Calibration Curves 14:31 Tbehteerc,oefficient of determination value for the calibration curve mustbe0.990 or 3M Enirommental Laboratory AnsysofScrEurmsExaisri2s Using ESIMS Pagel Page 32 3M Medical Department Study: T-6295.25 418-018:PAGE 1-426 Analytical Report: FACT-TOX-169 LRN-EO1-0180 14.3.2 Atlhle aecxtcievpetciaolniobfrattihoenLcOurQvepopionitn,tswhmiucsht mbaeywidtehviinat2e5u%potfo t3h0e%t.heoretical value with ` 14.3.3 Calibration standards with peak areas les than two times the curve must be deactivated to disqualifay data range that may be affected bmyatbraicxkbglraonuknd levelsofthe analyte. 14.3.4 Low or high curve points may be deactivated to optimize u linear range appropriate to the data. 14.3.5 `Nmoaty ibneclduedaicntgivlaotwedorifhtihgehypdoeivnitastedrmooprpeedthtaono2p5ti%mifzreomthtehleintehaerorreatnigcea,l cvaulruvee wpohiennts the curve is evaluated ovaelinrear range appropriate (0 the data. 14.3.6 OhanseapopienatkbaerlcoawletshsetLhaOnQtwmoaytirmeemsatihneamcattirviexebvleannki;ihtodweevvieatre,sthmeorLeOtQhanwi=ll3b0e% or defined atthelowest point with acceptable deviation. 14.3.7 A valid calibration curve must contain at least active points above and including the LOQ. 14.4 Matrix Spikes 14.4.1 cTohneceanvterartaigoen.maRtericxovsepriikeespoeurtcseindteorefctohviserireasngsehosuhlodulbde bweitdihsicnus3se0d%oinftthheerespporitk.ed 145 Continuing Calibration Verifications (CCVs) 14.5.1 pCeorncteinntuirnegcocvaelirbyrwatiitohninsa2mp5l%esofwitthhienstphiekeldinceoanrcreanntrgaetoifont.hIefraunCmCuVstissohuotwsiade of tbhriascrkeectoevderbyy,tshuebcsuerqvueenatnddaptaasssihnogulCdCnVo.t be accepted. Acceptable data must be 14:6 Ipfecrrfiotrermieadliosntetdhienstyhisstemmetahnoddspaemrpfleosrmraenacnealsyezcetdioonriosnt'htermeatc,timoanisnatsendaentceermmianyedbbey the analyst. Document all actions in the appropriate logbook. 14.7 1fdooattnaoatreed o1n0 tbaeblreespoarntdeddiwshcuesnsepderifnotrhmeatnecxet ocrfittehreiarehpaovret.not been met, the data must be 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 pSiapmepttleeweaxstrtaectiswdaisstpeosaenddifnlbarmomkaebnleglsaoslsvecnotntiaidniersspolsoecdatiendhiinghthBeTlUabocroanttoariyn.ers, and glass 16.0 RECORDS 16.1 Tihnfeorfmrasttipoangienocfluedaecdheidtahtearpianctkheethgeeandeerr,aitedtfhoerfaoostteurdyormuhsatndhawrvietttehne ofnoltlhoewipnagge: study or project number, instrument, sample matrix and tims point, date, and analyst. 16.2 `Acodmatpaoupnadckseutmimnaclruyderseptohretffoolrleowaicnhgt:ardgaettaanraelvyiteew,squuamnmtairfyy cfaolrimb,raMtiaosnsrLeypnorxtqfuoarnteiafcyh target analyte (curve), method report, Word document isting set-up parameters, tune. SM Environmental Laboratory Analysisof SerEumTSE3xa5c2t Using ESMS. Pages of 11 Page 53 3M Medical Department Study: T-6295.25 418-018:PAGE 1427 Analytical Report: FACT-TOX-169 LRN-EOL-0180 qmueatnhtoitdyrseapmorptl,eMraesposrLty(ncxhrsocmaantnoignrgamm)e.thod report, HPLC method report, sample lst, and { 16.3 Fpaorraemaetcehrsan,alMyasisss,Layfnexrspcrainnntiinnggtmheesthuonde mreeptohrto,darnedposrat,mpWloerldst,doccoupmyeanntditsatpineg,intsoett-huep instrument runlog. The original is maintained inthe data packet. 16.4 O`qnuanetaicfhy psaamgpeoleftrhepeorqtua(ncthirfoymcaotmogproaumn)d, rtehpeofrto,llqouwainnigifiynfcoalrimbartaitoinonmuresptorbte(icnucrlvued),edaenidther imnetthheodh,eacdaelri,brtahteiofnoo(ttehre,moerthhaonddawnrditctaelnibornattiohne apraegeu:suasltluydyasosrigpnreodjetchtensuammbeern,amiens)t,rument, analyst, and date. 16.5 aTrheeelaencatlryosnticmaulslty diantceluadnedd oinniticaalcthhepafgires.t Ipfaigneitiianlas apnadckdeatteasarleonngo:aseltehcitrroinniictailallsyainndclduadteed, they mast date and inital each page. 16.6 `SAutmtmaacrhimzeentdaAtfaoursainngesxuaimtapblleeosfoft2wsaurmem(ac.rgy.,sEpxrceeald)shfeoert.inclusiion the final report. See 16.7 Bbaacckkuuppeleelcetcrtornoincicdadtaataintoinasptprruompernitatleogmebdooiku.m. Record the file name and location of 17.0 TABLES, DIAGRAMS. FLOWCHARTS. AND VALIDATION DATA 17.1 Auachment A: Data summary spreadsheet 18.0 REFERENCES 18.1 cFAoCmTp-oMu-n4d.s1f,ro"EmxSterarcutmiofnoorfAnPaoltyassissiUusmiPnegrfHlPuLoCr-oEolcteacntersouslpfornaayt/eMaosrsOtShpeecrtFrolmueotrroyc.hemical 18.2 EToTnSi-z9a-t2i4o.n0/,Ma"sOspeSrpaetcitornomaentderMQauianttternoanIcIeotrfiptlheequMaidcrruopmoalsesSyAsttmesmpsh"eric Pressure 18.3 The validation report associated with this methodisETS-8-4.0 & 5.0-V-1. 19.0 AFFECTED DOCUMENTS 19.1 ECToS-m8p-o4u.n2d,s"EfxrtormacSteiornuomfPfoortaAnsasliyusmisPUesriflnugoHrPoLoCst-aEnleescatfroonsaptreaoyr/MOatshserSpFelcuotrroocmheetmriyc"al 531 Environmental Laboratory AnalysisofSeEruTmSE8x5ac2t Using ESIMS Page9a11 Page 54 3M Medical Department Study: T-6295.25 418-018:PAGE 1-428 Analytical Report: FACT-TOX-169 LRN-EO1-0180 200 Revisions ReNvuimsbeiro,n Reason For Revision 1 SSeeccttiioonn 61.11..12AvClearraigfiecoaftitonwoofcHuPrv!es1,00nostyssttaenmdacrodmpvoalnueenst,sare used for plotting linear regression and added the 1/x weightingof the curve. SSeeccttiioonn 1172..12:C2.h4anClgaerdififcraotmioantotfacshomlevnenttBratmop.A. 2 1100:2CAldadriefdieidnswthruecntibolnasnfkosrswrheernn.sumogat mati i ved 1101:13 SRpeeqcuiifryesreoquiraen cmleinbtesay toiroCnVcsu.rv befor the samples. 111132 ACllalroiwfyowhtartuntcoatdeotfhecucruvree.docs not mee requirments. 1122..1113 CCllaariiftyy tsyapmipcallerlusnt. ID. 12.2 Changes to mobile phase gradient and speityflow ae. 11443.33 ChWahnegentaocdceeapcttaibvlaetmeicsal.ibrAatdiosnpsetciafnidcasrfdosrbaacsceedpotannbceloafnkcarelsipobnasoe.n curve. 14.35 When todeactivate calibration standards within thelines range. 1144..54DMeosdcirfiibeesseevvaalluuattiioonn annddususeofofmCaCtVi. spikes. 1164.50.1AdEdvarleuqautiiroenmoefntCsCVfosr.records and documentation. ReDviitsieon 04/02/99 Auschment A: SummarySpreadsheet`AnalysisofSeruEmTSEx8t5ra2ct Using ESMS 3M Environmental Laboratory Page 100f11 Page 55 418-018:PAGE 1-429 : LRN-E01-0180 Laboratory Study # 5 am[er[oe[a|ar[vo | I 418-018:PAGE 1-430 TT Appendix D: Data Summary Tables. | a [rsJremermen ve|msm = = mir|reesrovsmrons| moon mmo | a m=e=| a | mee | w| wow goee| EE | = |e | HSsE || ww ww | ow | i=nGaps NA wo= 2724187 I 2:38 eaiiie|| ow || ow | wr | Gres sae 1 1708381 EEe t EE rr NNOOTTEE Risoasasoo troesvean.rytarc ofGnisnoptiossratasin scuss wrokroa id rncmarlTh ny messinf- 3M Medical Department Study: T-6295.25 04 Metical Deparment Suc: T-6295.25 Appendix E: Data Spreadsheets 418-018PAGE 1.431 Analytical Report: FACT-TOX-169 LRN-EOL.0180 Anaytal Ror: F(ARCNT.T0O1X.-0116890 $M Environmental Laboratory Page 38 | 418-018:PAGE 1-432 EHIHH == =EE [e TIET=TeE ET rm]ES] EERE eT.| =EiE EfTE .T.] EE EEE TT] EE EEE 1.TL] 418-018:PAGE 1-433 A yuSR ie Ee, EES Arwen re frei Ei | Se n prc EEE) EE = HEE PEE: i TEE fli E EEl EE E lLl [L] Ef Z| iEE ||, ee mss BE cements EVEIOIS0 |= =EE [E ETiEeTEETEL --l ETEE EEEE L] 3M Environmental Laboratory Page 61 SM Medial Dparmen Sy: T9525 fu pron somoomomeeEEE 418018PAGE 1435 Avalyca Report: FACT-TOX 169 ps sit i reer I lv l = = (Ei[ifEE EL. SEs Pages? 418-018:PAGE 1-436 3M Medical Department Study: T-6295.25 aM Mecical Deparment Stuy: T-6295.25 Analytical Report: FACT-TOX-169 LRN-EOI-0150 Anaytcal Report FLARCNT.T0O1X.-0116890 Appendix F: Example Calculations Formula UsedforSora Analyses in Study FACTTOX-169 AR (ng/mL) x DF x FV (mL) x 10ug = Reported Concentration (ug/mL) EV(ml) 1000ng Calculation Used for Group 3, F9, GD21, Animal ID 10830F 530ng/mLx400x O130mmLL x T1000p0ngg = 70.7 pg/mL DARF-- DAinlaultyitoincaflacrteosurlt from MassLyn summary [EFVV----_TFVionlalumexetorfactsevroaleuxmtreac(1t.e0d mL unless otherwise noted) $3 Environmental Laboratory Page 63 418.018PAGE 1437 3M Medical Department Study: T-6295.25 a0 Mec! Deparment Suc: T-6295.25 Analytical Report: FACT-TOX-169 LRN-EOI-0130 Analytical Report F(ARCNT-T0O1X..0116890 `Appendix G: Interim Certificate of Analysis $3 Enironmental Laborators Page 64 418-018:PAGE 1-438 3M Medical Department Study: T-6295.25 Analytical Report: FACT-TOX-169 . | LRN-EO1-0150 - Centr30e48 RAasneaarlehyDtiviocal Laboratories, Inc. Stale Cale. PA 16201 . Prone: (814) 2318032 Fax: (514) 231-1253 or 814) 2311580 INTERIM CERTIFICATE OFANALYSIS Centre Analytical LabRoervaitsoiroines1(C9/O7A/00R)eference #: 023-01A ` ema Ee 3M RPreofdeucrte:n#cPe:FOSSD,-L0o1t8 217 Purity: 869% seetes T Rew 1 Ew] Cons | TaentfeNstRon el1s. (CaClPciSu)m 23.. MSoadginuemsiom 456.. PNIoiotcnakseslium' 7._ Manganese otal %Impurity(NMR) . |"aTeoumls)lmpury| (ReGaetnesd) Compound--s PReOsiAdAual Solvents (TGA) 2L. 00000015wwaislne 5 L439wen 465.. 6<0.0.800400981wwwrisiwmtn% 7_<0001 wis NoweDetected T | T0o3n3ewDeitmected Not Applicable T|norg21a.niCcFhAolnoinnodenes (IC) 2L 0o0s1o5wwiemns | | 555 NbNreiomeeide 5350: 0<Z00o000a906wwwiemlwitsk|| | 76 Pshoopehe || i 75 ._<0803076wwlnhd |1 1 EA 2. PFPA | 1 <0lwmn 2. <01wim | 5 mma | a wea 3 0l0wins 5_028wem% [| ElememaClarAbronaya I TheoreticalValue 176% L 1248 wth | 25. NHyidorgoegnen 25. ThTehoeroreettiiccaallVVaalluuee ==00%% 32. 012s7wi4n " | 3 sume 5. Fluorine ! 5 TheoreiealVabes 595% S. Theoretical Value=60% || 4 5. S84wein Salwtiw% | cosomaisa 3M Environmental Laboratory Page tor Page 65 418-018:PAGE 1-439 3M Medical Department Study: T-6295.25 i Analytical Report: FACT-TOX-169 LRN-EOL-010 i | Centre Analytical 3048 Research Dive Laboratories, Inc. Site College. PA 16801 Phone: (314) 231.8082 Fax: (814)2310-r(6114)22015-15380 INTERIM CERTIFICATE OFANALYSIS Centre Analytical Laboratories COA Reference #: 023-0134 DateofLast Analysis: 0831/00 Expiration Date: 08/31/01 Storage Conditions: Frozen 10C Re-assessmentDate: 08/31/01 "Purity = 100% - (sumofmetal impurities, 1.45% +LC/MS impurities, 8.41%+Inorganic F0l3u3or%i)de, 0.59%+NMR impurities, 1.93%+organicacid impurities, 0.38%+POAA, TotPaulriitmypu=ri1ty0f0r-%om1a3ll.t0es9t=s%=8163..90%9% Potassium is expected in this salt form and is therefore not considered an impurity. opbusreirtvyebdyfoDrStCisissgaemnpelrea.lly not applicable to materialsof low purity. No endotherm was "iSnuolrfguarniicn atnhieosnammeptlehoadppceoanrdistitoonbse. coTnhveeratneidontoreSsOul4t aangrdeheesnwceeldletweictthetdhuessiunlgfutrhe determination in the elemental analysis, lending confidence to this interpretation. Based on the results, the SO, is not considered an impurity. SHTFFABA `THreipftlaufolruooarcoebtuitcyarciicdacid PNFFPPAA PNeonntoafflluuoororporpoepanntoaicnaocaciicdd "Theoretical value calculations based on the empirical formula, CF17SOSK" (MW=538) "This work was conducted under EPA Good Laboratory Practice Standards (40 CFR 160). coxomatsa $3 Environmental Laboratory Frgeon2 Page 65 418-018:PAGE 1-440 3M M.edial Department Study: T-6295.25 Analytical Report: FLARCNT--ETOOIX--0116590 Ceno trPheonw e.A(n51a4R )l2y31t8e 0i3c2 as lFaxL:ae (8b14o) 2ra 3a1t-S1ao2O wr5C0o31il(ee5gm 1se4),2P3A1I-e IG1n8I8cO0T. CenItNreTAEnaRlyItiMcalCLEabRoTraItoFriIeCs CAOTAEROefFereAnNceA#L: Y02S3-I0S184. LOMS Purity Profile: o [i CTo7ur W1E32E [1533 1 aBam t Ncruoortvmee:st,hTerheCesp4Cec4atniavdenCldy6.C6sTtvahanelduaCerSsd wcvuearrvleees,cwaalLcsiuklcaeatwleicdsuleua,sttiehndegtuCsh7ienCgv4atlahueenvweCadr6sagcsaetlacrnuedlsaaprtodendsceuaslfiiabncrgtaottrihsoen average response factors from the C6 and C8 standard curves. ( Prepared BCy: raASBgellorel ABueke Reviewed By: Sci(cZLp.i1st3, CeFpltozy Aghlytcal Laboratories ZZ ohn Flaherty Date Laboratory Manager, Centre Analytical Laboratories cosomaisa 3M Ensironmental Laboratory Pesan Pages? 3M Metical Department Study: T-6295.25 3 Metcal Deparment Stuy: 6295.25 Appendix H: Report Signature Page 418-018:PAGE 1-441 Analytical Repors FLARCNT--ETOO1X--0116590 Anica Report: FRAeCTorTOoXt-1e600 lovead ody Deanna J. Luebker,\1.S., StudyDirector lens Date Poses TG Marvin T. Case, D.V.M., Ph.D., Sponsor Representative 6 Duns Dr Cate Keston A J. Hany sen, P/ h...-- PrincipalAnalytical mvesigator LeDnaeto 2001 Y Wiliam Kr . Reagen, Ph--.0., (aboratoryManager osGab te y $M Emironmentl Laboratory Page 65 418-018:PAGE 1-442 PFOS Concentrations in Rat Livers Souther Research Institute Study A344.1.4 418-018:PAGE 143 Methods W1:e5 (fbirystwperiegphatr)ewdictohntDrIolwattiesrs.ueThhoimsowgaesnautseedbytohpormeopgaerneitzhiencgalriatbrlaitvieornfsrtoamndcaorndisr.ol Wraets salpiiqkueodteisnokfntohwenblaamnokunttisssuoeftheosmtogarnteiactlee.(PAFObSl)anaknd+ iinntt.esmtad.l astnadndaabrlda(nPkF-OiCn)t.isnttdowere prepared with each calibration curve. `aTnhyeofstamhpelsees swaemrpeleaslswoehroemwoigtehniuznsepdi(k1e:d5)blbaynwkeciognhttrowlitlihveDrIhwoamtoegr.enaDitleutpiroenpsarmeaddaebotvoe. "Then an equal amount of internal stanard (PFOC) was add to each sample. "Then to both the samples and carbonate/bicarbonate buffer, standards we added | mLof DI water, and S00 uLof ion pairing reagent (0.5 1 mLof M 0.25/0.25 M tSoelturtaibonutwyalsammmioxneidumbrhiyedflryoxaindde)thaedwjuestaedddteodp2H.51m0.L5 owfitehthsyoldaicuetmathey.drTohxiidsem.ixTthuere was csehnatkreinfufgoerd|athr25o0n0arpplamtffoorrmS mmiixn.er.ThAefteetrhyaln ahcoeutrattehelasyaemrpilsresemaorevetadkeannodfefvaanpdorated 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. S`tSuoduythAe3raR4e1se4arch Insitute PFOS in Rat Livers FiloName_ Sample Namo. Measured Cone. (ngiml) 65//2288//22000045 FFOO 1100990068ccoonnttrrooll 51558523 6600228822000076 FFOO 1110580113ccoonnttrrooll 032.478835 6610228822000089 FFOO 1111550045ccoonnttrrooll 11508259 660022881122001110 FF11 1100990068ccoonnttrrooll ""91.020678 6282012 6282013 FF111110590111ccoonnttrrooll 8a50z4 66/22882/2001145 FF111111550032ccoonnttrrooll 1"012075 661228822001178 FF11110100..9044 mm90gkk80g 2153775 660228822001219 FF11111106001000..44mmgglhkkgg 8138947 660228822005242 FF111116012060..44m0mogk1kag a2r1s5 66228822002245 FF1 11111116..066 4mm4gah0h9kag 4112475 66228822002268 FF1 11111116..666 4mm4aa1kk2aa e051399 2628822002390 FF11111111..6666mm44yghh84kkgg 8052071 660228822003323 FF111111065594 22mmoghkkag 8362023 660228822003364 F11111166555 22mmoglkkag 0864552 662288122003378 FFOO 1111600004 00..44mmggllkkgg 38s18022 55228822004410 FFOO 111166003100..44 mmhgg 105855 5282043 6282044 FFOO1111664403 11.66mmggikkgg 193221 51282045 60282047 FFOO 111111665644mmgo25kkga 98190.245 66228822004580 FFOO 1111664569 12.m6mgohlkkgg 96134829 5282051 6282052 FO FO 1111666547 22 mmaglkkgg 11109857 5600228822005535 FO0111165082 2m5gmhg9khgg 11038505 61282056 FO 1162m5gh4kg 1182 "Pup and Dam samples were mixed together during preparation 80L = below quantitation imi, < 12.5 ng/mL. OR nglg CCoanlec.ul(antgeidg) ssaall BBaall 21.020815 ssaal ssaar ssaall 9203.77435 140313,71582 7918..042279 111961..870502" 714786.89985 324092.309880 315463..126392 218919.,846650 5248.82041 4458550303 112322.24300 1873.02815 112581 115509..003145 113831..813711 110.148 161501 418-018:PAGE 1-444 APPENDIX J HISTOPATHOLOGY REPORT 418.018PAGE J-1 RESEARCH PATHOLOGY SERVICES, INC. 438 EPahsotneB:utl2er1A5v.en3u4e,5.N7e+0wF7aBx0r.ain2,16.P3A451.84930216 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.AB.T. Associate Director of Research Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 SUBMITTED BY: WR Brown Dec 13, 2002 W Ray Brown, D.V.M., Ph.D. Veterinary Pathologist December 13, 2002 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 RESUS 3 SUMIRY....ccorrrer sso SE ---------------- 4 Quality Assurance Unit Statement 5 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. EI =I I [wom [T|o orn m | te ome | Cem= |=Towrn|owe| oa on, | C rs l =Toawn, |i www |e JEooTn, | CleT=Toon|vers | me, | Com T=Tom| new| =| Come To[oom |oe| = HTEomer |EaEToE Tourn|] CCeoTomimuerrssT|o0T TnT|owernn ||e| ] CCooTommaom TTaoT Tu u|Tummurss| | w] ] Co TomomseTatt| er | mere|] 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 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 liter) or Day 4 postpartum (rats that delivera litter). Rats in the process of delivering were not given the test/control article. Male rats were used only as breeders and were not considered part of the test system. F1 generation pups were not directly given the test article, but were possibly exposed to the test article during maternal gestation (in utero exposure) or via maternal milk during the lactation period. Dosages of the FO generation rats were adjusted for the most recently recorded body weight and given at approximately the same times each day. On Day postpartum, pups were sacrificed and the heart and thyroid were collected and preserved in 10% neural buffered formalin. The in-ife portion of the study, necropsies, and recording of the gross observations at necropsy were performed by the staff of Argus Research Laboratories, Inc Representative sections of the heart and thyroid of one male and one female F1 generation pup from the vehicle control group (Group 1) and 2 mg/kg PFOS group (Group 13) were routinely processed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin for microscopic examination. Upon completion of the. project, all raw data (remaining wet tissue, paraffin blocks, microscopic slides and histology records) will be returned to Argus Research Laboratories, Inc. for archiving. Research Pathology Services, Ic. 2 418-018:PAGJ-E5 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. SDeoxse Group: DNuammbNerumobfePrups Examined: TNoH.YREOxIaDm:ined No. Normal HNoE.ARETx.amined No. Normal M7 om7 10901 9 1101 59 11 11 11 11 7F [F 109109 110159 11 11 11 11 No microscopic changes were observed in the heart or thyroid of the male and female pups exposed to 2 mg/kg of PFOS. The sections examined were all within normal histologic limits Research Pathology Servis, nc. 3 418-018PAGE J-6 ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACIDICHOLESTEROL 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 female pups exposed to 2 mg/kg of PFOS. Research Pathology Services, nc. + 418-018:PAGE J-7 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 QUALITY ASSURANCE UNIT STATEMENT report prepAalrlataisopnecftors tohfetshteutdiyssluiestepdroacbesosviengh,avmeicbreoescnoppiecrfsoirdmeedpraecpcaorartdiionng, tohitshteopSatthaonldoagridc Oevpaelruaattiinogn Parnod- `bcyedtuhreesQoufalRietsyeAasrscuhraPnactehoUlnoigtyoSferRveisceesa,rcIhnc.PaatnhdolwoegryeSaeurdviicteesd,inInacc.cionrdcaonmcpelawnichethweitphrtohceedGuoroesd eLsatbaobrlaitsohreyd Practice regulations specified in the protocol. SnteudoynDedestiegcntiaon MofoxdiccaFiooornfe.pUroSdFucotoidonafonrdmerdginAadpiriasct.aFta1d9oo9r4na)l.Rotgeirstaatro,nSCeoptnef2mo,eb1ne9rK8ar,meaVnnoi.s5ca6o,enN;o.Gu18d3e: US Foodan DrugAdriaan, GoodLaboPrractacstReogironys: Final Rue. 21CFRPan 8 `ECuotmompaenaintyEocforaarOiEcCCoDmdmeiiatnf(u1c8a8m9i)n.aCnoduantodndaoncicsomopnloa2nceuwliy 1p96r9i0 oheoafcgcoptdanccreabtoynrecEutreo.peOafnfiEccoramiucra fhe Etopean Communes: Logaton. 32 No. L328O1ct5o: 117. JnaapnacneeseNuMmibserty21o, Meaarclh 2a5d,1W9e9l7l (1657). GuodLabaratyPrctieStandardforSaytusonDrugs,MANO. tions QfuraolmitythAesspruortaoncocle, USntitansdtaurddy-ObpaesreadtiinngspPecrtoicoendsurweesreanpdelrofrorampepdroapsrisatheowGnoobedloLwa.borTahteorreywPerraectnicoederveigau-- plaotritosnsubnmoitteedddutroitnhgetShteucdoynDdiurcetctoofrtohne Dsteucdye.mbTehre13s,u2m0m0e2.ry report of GA inspections is included in the fina re- InDsapteescioofn Study Phase DatMeasnaRgeepmoernttetdo tDoaStteudRyepDoirrtecetdor o0a4n00B9i0011 MParset-enrtSicathieodnule 0o4a3r0300011 1122111330022 o0a5r021a1i0011 ETmrbiemdmdinigng 00543310/00011 112211113300022 0068110087110011 MiSctraoitnoimngy 006613300110011 11221111330022 oo77nn7ao0tt HistDoaptaatEhnotlroygy 0773311110011 1122113300022 0077227500011 DDaattaa VPerroicfeicsastiinogn 0773311110011 11221113300022 007830030011 PRreop-osrutbmPirsespiaornatAiuodnit or08a1t08o10r1 1122111330022 08080112,011300/217/01 DFrianfatl rreeppoorrtt oB0B1O211,1310021701 112210113300022 A bey J Mhkins DD KareQhualWi.tyHAasrskiunrsa,ncBeSUnit 2:13 02 Date Research Pathology Services, Inc. 5 APPENDIX K| HISTORICAL CONTROL DATA SUMMACRrYLCODF(SRD)EIPGRSOBDRUCRTAITVSE INDICES DAY 21 C-SECTION 418-018:PAGE K-1 PERIOD JUNE 1998 - SEPTEMBER 1999 NUMBER OF STUDIES 3 NUMBER OF RATS: TESTED 252 PFROEUGNNDADNETAD 235 1 ABORTED 0 DELIVERED 0 NUMBER OF RATS PREGNANT AT CAESAREAN-SECTIONING 23 NUMBER OF RATS WITH SINGLE CONCEPTUS LITTER: uve 0 RABEOSROTREBDED 0 0 % PREGNANT AVERAG#E CORPORA LUTEA AVERAGE # IMPLANTATIONS AVERAGE LITTER SIZE AVERAG#E LIVE FETUSES AVERAGE # DEAD FETUSES AVERAGE # RESORPTIONS AVERAGE # EARLY RESORPTIONS AVERAGE # LATE RESORPTIONS MEANOr% 934 178 149 145 [4 [3 05 00 RANGE/STUDY MEAN Or% (67.5100) (16.4191) (14.0183) (13.659) - 011.4) 11.4) 0.1) 418-018:PAGE K-2 `SUMMACrREYCOD(FSRD)EIPGRSOBDRUCRTAITVSE INDICES DAY 21 C-SECTION AVERAGE % DAMS WITH ANY RESORPTIONS AVERAGE % DAMS WITH ALL CONCEPTUSES RESORBED AVERAGE % DAMS WITH ONE OR MORE LIVE FETUSES AVERAGE SEX RATIO, (% MALESILITTER) AVERAGE FETAL BODY WEIGHT (6) AVERAGE FOR MALES (6) AVERAGE FOR FEMALES (6) AVERAGE % DEAD OR RESORBED CONCEPTUSESILITTER MEANOr% 393 00 1000 94 528 5.40 512 33 RANGE/STUDY MEAN Or% (125714) - - (43.0:57.0) (5105.48) (5155.63) (492529) (0888) `SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CrtCD(SD)IGS BR RATS - DAY 21 C-SECTION 418-018:PAGE K-3 PERIOD JUNE 1998 - SEPTEMBER 1989 # STUDIES 13 # RATS TESTED 252 # RATS PREGNANT 235 # RATS DIED 1 # RATS ABORTED 0 # RATS DELIVERED 0 # RATS WITH 100% RESORPTION 3 OBSERVATIONS ver Left lateral lobe, tan area UTERUS Moderate hydrometra INGUINAL AREA Subcutaneous, tan mass. onright side RANGE/ STUDY NO% ON % 1040 01 (040) 1040 01 (042) 1040 01 (048) SUMMCrAiRCYD(OSFDF)IEGTSABLRGRROASTSS -EDXATYER21NAGL-SAELCTTEIROANTIONS 418-018:PAKG-4E PERIOD. JUNE 198 - SEPTEMBER 1999 # STUDIES INCLUDED 13 # LITTERS EXAMINED 23 # LIVE FETUSES EXAMINED 3383 ALTERATION EVES) Eyebugesdepressed Jaws Micrognathia HINDPAWExtra digitpresent TAL Threadiike No% L 1 043 Fo1008 RANGE /STUDY ON % 01 (042) 01 (003) L104 01 (042) F100 01 (003) FL 1100408 0011 ((0040835)) L104 01 (043) F 1003 O01 (003) L: LITTER INCIDENCE F: FETAL INCIDENCE 418-018:PAGE K-5 `SUMMARY OF FETAL SOFT TISSUE ALTERATIONS. CrECD(SD)IGS BR RATS DAY 21 C-SECTION PERIOD JUNE 1998 - SEPTEMBER 1999 # STUDIES INCLUDED 8 # LITTERS EXAMINED 180 # FETUSES EXAMINED 1245 ALTERATION BRAIN Dilofalatetralvientoricnles L Fo RANGE/STUDY N% ON % 2 1.11 0:2(083) 2 016 02(012) HEART Septaldetect LFo 1100808 0011(004028)) VESSELS Umbilicalartery, descends toleftof urinarybladder innominateartery, absent Aonapadorssasl teotshe tracheaandesophagus Lfefitgtchaehrrriisgteohstttcoaoirtohgfteid Rifgghhttocafrtohteirdirgihsiessutbo-the clavian Pumonary artery, small Semiunarvave, small Ductus arteriosisattached totheleftsubciavian ~~ Great vessels, transposed L 3 1.67 F 3 024 L 2 1.11 F206 L 1 056 F 1 008 L11 00.5068 L106 F 1 008 L 1 056 F 1 008 LFo 11 056 008 L 1 056 F 1 008 LF 22 1.11 016 02 (09.1) 02(012) 02(083) 02012) 0-1 (04.2) 0-1 (0:08) 00--11((00--4026)) 01(0:42) 04(008) 0-10-42) 041008) 0-1(0-42) 00-11((00-0482)) 0-1 (0:08) 0-1 (0-43) 01(007) LUNGS Apicalandcardiac lobes, L absent F 1 056 0-1 (0-42) 1 008 01008) KIDNEY(S) Pelvis,slight dation Pelvis, marked dilation Lo 1 088 01(045) F 1 008 01(007) L 1 056 0-1 (0-45) F 1 008 04(007) L: LITTER INCIDENCE F: FETAL INCIDENCE 418-018:PAGE K-6 `SUMMARY OF FETAL SKELETAL ALTERATIONS CriCD(SD)IGS BR RATS - DAY 21 C-SECTION PERIOD JUNE 1998 - SEPTEMBER 1999 # STUDIES INCLUDED 8 # LITTERS EXAMINED 180 # FETUSES EXAMINED. 1332 SKULL ALTERATION Eye socket: small Mandbles: short VERTThEorBaRciAcE: Centrum,bifid aCreenntrum,fusedto 4present Lumbar: Fused + 8present Centrum,fusedto ~~ arches RIBS CervicalRib(s) present. Oneor more,wavy Oonsesiofremdo(rhey,poipnlcaostmipcl)e,teolry.not ossiied Fused 4present No% L108 Fo1008 L 10% F 1008 L422 F 403 FL 11000586 FL 11000s8 LFoo22 011115 L 10% F 1008 L 1 056 F 1008 L 12 667 F L 1a531035 F 8 060 L211 F 302 FL 11000%8 L108 F 1008 RAN/SGTUEDY Nw 01 (042) 0011 ((000428)) 01 (008) 01 (048 01 01 ((000488)) 01 (008) 01 (048) 01 (008) 01 (048) 0011 ((000488)) 01 01 ((00-0488)) 01 (008) 05 (0208) 0062 ((003955)) 03 (018) 02 (09.1) 03 (018 0011 ((004028)) 01 (048) 01 (008) L: LITTER INCIDENCE F: FETAL INCIDENCE 418-018:PAGE K-7 SUMMARY OF FETAL SKELETAL ALTERATIONS CriCD(SD)IGS BR RATS - DAY 21 C-SECTION MANUBRIUMALTERATION Duplicated STERNEBRAE Oneormore incompletely ossiied or notossilied Asymmetric Duplicated XIPHOID Duplicated HINDLIMB Extra digtpresent Femur, bent RANGE STUDY No% ON % FL1100088 0011 ((004058)) L422 F 4030 LFoo22 011115 FL 1100058 02 (09) 02 012 0011 ((004088)) 0011 ((004058)) FL 11000s8 0011 ((004058)) L108 Fo1008 FL 11000s8 01 (048) 01 (008) 0011 ((000488) L: LITTER INCIDENCE F: FETAL INCIDENCE SUMMARY OF FETAL OSSIFICATION SITES. C`rS.KCEDL(ESTDA)LIGASVBERRARGAETSS DAY 21 C-SECTION 418-018:PAGE K-8 PERIOD: JUNE 1898 - SEPTEMBER 1998 # STUDIES INCLUDED 8 # LITTERS EXAMINED 180 # FETUSES EXAMINED 1831 `SKELETAL AVERAGES HYOID VCEERRTVEIBCRAALE THORACIC LSAUCMRBAALR CAUDAL RIBS (pairs) SMTAENRUNBURMIUM XISPTEHRONIADL CENTERS FOREPAWS (Calculated as average per imb) CARPALS METACARPALS DIGITS PHALANGES HINDaPvAerWaSge(Cpaelrcumlbat)ed as TARSALS DMIEGTIATTSARSALS PHALANGES FETUSILITTER MEAN RANGE/STUDY 089 (096.00) 7.00 1807 (13-034314) 592 (585597) 300 - 178.0454 ((1731012716391)2) 100 - 319090 (398-4.00) 000 - 400 (399-400) 500 - 828 (804852) 002 (0007) 45.8000 (47-3487) 621 (581654) APPENDIX L STATEMENT OF THE STUDY DIRECTOR 418-018PAGE L-1 r~PriPMREIMETICA pa Ar5g05RSehsecahrychDrLiavbeo,raBtuoirlideisngIncA TelTephcon:e: 2(21139) 444433-85578107 PROTOCOL 418-018: ONE GENERATION REPRODUCTION STUDY OF PFOS - MEVALONIC ACID/CHOLESTEROL CHALLENGE AND NOEL INVESTIGATION IN RATS SPONSOR'S STUDY NUMBER: T-6295.25 STATEMENT OF THE STUDY DIRECTOR "This final report accurately reflects the raw data obtained during the performance of the study. No deviations from the U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations; Final Rule', the Japanese Ministry of Health and WtehelfOarrgean(isMaHtiWo)n GfooroEdcoLnaboomriactoCroy-oPprearcattiicoenSatnadndDaervdefloorpSmaefnetty(SOtEuCdDi)es.onThDeruRgesv"isaendd OECD Principlesof Good Laboratory Practices' occurred that affected the quality or integrity of the study, with the exceptions in the bulleted list bellow: There were two study directors for this study; one for the in-life phase and one for the analytical phase. The in-life study phase was considered to end a 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. Cerne 0A eae Rayolond G. York, PED, DABT Date Associate Director of Reschrch and Study Director Argus Research Laboratories, Inc. a. US. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standardfor Safety Studies on Drugs. Ordinance No. 21, March 26, 1997, Japan. . Organisation for Economic Cooperation and Development (1998). The Revised OECD PrinciplesofGood Laboratory Practices [C(97)186/Final). APPENDIX M QUALITY ASSURANCE STATEMENT r+,PriFRIMEDICAA 418-018:PAGE M-1 LE Arg"u9s03RSeisceeahrych DLraibvoe,ratBourilideisn,gIcA TeleTpelheofnaex:: (22115)54)4434.38-8578170 QUALITY ASSURANCE STATEMENT Argus Protocol: 418-018 Study Director: Raymond G. York, Ph.D. DABT Sponsor's Study Number: T-6295.25 "The protocol, critical phases, raw data and final report were inspected by the Quality Assurance Unit (QAU), to assure conformance with: USS. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule 21 CFR Part 58. Japanese Ministry Studies on Drugs, of Health and Welfare (1997). MHW Ordinance Number 21, Good March Laboratory 26, 1997. Practice Standard for Safety EhueroEpueraonpeEacnonEocmoincomCiocmmCuomnmiutnyi(t1y98o9f)a.nCOoEunCciDlddeecciissiioonn/ornec2o8mmJuelnyda1t9i8o9noonn tchoemapclcieapnrcaenwcietbhy principlesofgood laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. `pTrhoevuidnedderbsyigtnheedSipnodniscoatreotrhaatstuhbecornetporratctiosranwearcecunroatteaurdeiptreedsebnytatthieonPorfitmehdeicraawArdagtuas. QuDaaltiaty Assurance Unit. sTthaeteddraafbtovper,otoonco2l7foJrUtNhis00s,tu3d1yJwUaLs00au,di1t5edAfUorGad0h0earnendc2e1toAtUhGe a0p0p.ropriate regulations(s), as `The QAU inspection and report audit dates are listed below: Inspection Phase Date(s) Findings~~ Date(s) Findings Submitted to Study Submitted to Inspection Date(s) _ Director Management Test Article Preparation Blood Collection Caesarean-Sectioning Tissue Collection Natwral Delivery/Liter DanvLitter Sacrifice Milk Collection 04 OCT00 310CT00 310CT00 310CT00 02 NOV 00 06NOV 00 06NOV 00 110CT 00 310CT00 03 NOV 00 03NOV 00 03 NOV 00 08 NOV 00 08 NOV 00 110CT00 310CT00 03NOV 00 03 NOV 00 03 NOV 00 08 NOV 00 08 NOV 00 Observations Inspection Phase In-Life Raw Data Formulations Raw Daa Report Tables Report Text Revised Report 418-018:PAGE M-2 Inspection Date(s) Date(s) Findings Submitted to Study Director Date(s) Findings Submitted to Management 1189JIAANNOOII,, 22JANOI to 22S691IIJAAANNNOO0II1 0and 01 FEB01, 000957 FFFEEEBBB 000111,0and 12 FEB OI (0 219SFFEEBBO0L1to 28 FEB 01 25 FEB 01 to 2236 MFEABR01OIa(n6d 29 MAR 01 17 MAR 01, 30 MAR 01 and 02APR 01 to 03 APR 01 to 10 APR 01, 12 APR OI, 13 APR 01 and 16 APR01 and 17 APR 01 18 APR01 04 MAY 01 and 0177MJUALYOI01 100CT01 300CT 01 31JAN 02 04FEB 02 27 MAR02 10 DEC 02 01 FEB 01 19FEB 01 01 MARO1 29 MAR 01 03 APR 01 16 APR 01 17 APR 01 18 APR01 07 MAY 01 17JUL01 100CT 01 310CTO01 31JAN02 04FEB 02 27 MAR 02 10 DEC 02 01 FEB01 19FEB01 01 MAR 01 29 MAR 01 03 APR 01 16 APR 01 17 APR 01 18 APR01 07 MAY 01 17JULOI 100CT01 310CTO01 31JAN 02 04 FEB 02 27 MAR 02 10 DEC 02 DB sso 13507 Mafthew J. Vaneman, B.S. Date Manager, Regulatory Compliance o faw Chea n te erecn ea "Cynthia M. Kelsch Date Quality Assurance Associate and Principal Auditor