Document 2Nj66eMB2RVLLz26LbOaXVmq7

DownloadViewSend us a messageRandom document
A R r ) & -o & 5 FINAL REPORT PROTOCOL 418-013 ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.12 FINAL REPORT DATE: 24 JUNE 1999 00780 PROTOCOL 418-013 ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.12 TABLE OF CONTENTS SUBJECT I. SUMMARY AND CONCLUSION A. Methods B. Results C. Conclusion II. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study A.1. Sponsor A.2. Testing Facility A.3. Study Number A.4. Sponsor's Study Number A.5. Purpose of the Study A.6. Study Design PAGE 1-1 1-1 I-2 I-4 11-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1 i C007S1 SUBJECT A.7. Regulatory Compliance A.8. Ownership of the Study A.9. Study Monitor A.10. Alternate Study Monitor A.11. Study Director A. 12. Technical Performance A.13. Report Preparation A. 14. Report Review A.15. Date Protocol Signed A.16. Dates of Technical Performance A. 17. Records Maintained B. Test Article Information B.1. Description B.2. Lot/Batch Number B.3. Date Received and Storage Conditions B.4. Special Handling Instructions B. 5. Analysis of Purity C. Vehicle Information C.1. Description C.2. Lot Numbers C.3. Date Received and Storage Conditions C.4. Special Handling Instructions ii PAGE 11-1 II-2 II-2 II-2 II-2 II-2 II-2 II-2 II-2 II-3 II-3 II-3 II-3 II-3 II-3 II-3 II-4 II-4 II-4 II-4 II-4 II-4 000732 SUBJECT C.5. Analysis of Purity D. Test Article Preparation and Storage Conditions D.1. Sample Information D.2. Analytical Results E. Test System E.1. Species E.2. Strain E.3. Supplier (Source) E.4. Sex E.5. Rationale for Test System E.6. Test System Data E.7. Breeder Male Rat Data E.8. Method of Randomization E.9. System of Identification F. Husbandry F.1. Research Facility Registration F.2. Study Rooms F.3. Housing F.4. Lighting F.5. Sanitization F.6. Feed F.7. Feed Analysis Ni PAGE II-4 II-4 II-5 II-5 II-5 II-5 II-5 II-5 II-5 II-6 II-6 II-6 II-6 II-7 II-7 II-7 II-7 II-7 II-7 II-8 II-8 II-8 000753 SUBJECT PAGE F.8. Water II-8 F.9. Water Analysis II-8 G. Methods II-9 G.1. Dosage Administration II-9 G.2. Rationale for Dosage Selection II-9 G.3. Route of Administration II-9 G.4. Rationale for Route of Administration II-9 G.5. Frequency of Administration II-9 G.6. Length of Study II-9 G.7. Method of Study Performance 11-10 G.8. Gross Necropsy 11-11 G.9. Statistical Analyses 11-13 III. RESULTS 111-1 A. Mortality, Premature Delivery, Clinical and Necropsy Observations 111-1 A.1. Mortality and Premature Delivery 111-1 A.2. Clinical Observations MI-1 A.3. Necropsy Observations IM-1 B. Body Weights, Uterine Weights and Body Weight Changes MI-1 B.1. Precohabitation MI-1 B.2. Gestation MI-2 C. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values MI-2 C.1. Precohabitation iv III-2 000754 SUBJECT C. 2. Gestation D. Caesarean-Sectioning, Litter and Fetal Gross External Observations D.1. Day 15 of Gestation D.2. Day 21 of Gestation REFERENCES APPENDIX A - REPORT FIGURES Figure 1. Body Weights - Precohabitation Figure 2. Maternal Body Weights - Rats Caesarean-Sectioned on Day 15 of Gestation Figure 3. Maternal Body Weights - Rats Caesarean-Sectioned on Day 21 of Gestation APPENDIX B - REPORT TABLES Table 1. Clinical and Necropsy Observations Summary Table 2. Body Weights - Precohabitation - Summary Table 3. Body Weight Changes - Precohabitation - Summary Table 4. Maternal Body Weights and Gravid Uterine Weights Gestation - Summary Table 5. Maternal Body Weight Changes - Gestation - Summary Table 6. Absolute Feed Consumption Values (g/day) Precohabitation - Summary Table 7. Relative Feed Consumption Values (g/kg/day) Precohabitation - Summary Table 8. Maternal Absolute Feed Consumption Values (g/day) Gestation - Summary PAGE 111-2 111-3 III-3 III-3 III-4 A-1 A-2 A-3 B-1 B-4 B-5 B-6 B-10 B-12 B-13 B-14 v 0007SS SUBJECT Table 9. Maternal Relative Feed Consumption Values (g/kg/day) Gestation - Summary PAGE B-16 Table 10. Caesarean-Sectioning Observations - Summary B-18 Table 11. Litter Observations (Caesarean-Delivered Embryos or Fetuses) - Summary B-21 Table 12. Clinical Observations - Individual Data B-22 Table 13. Necropsy Observations - Individual Data B-29 Table 14. Body Weights - Precohabitation - Individual Data B-34 Table 15. Maternal Body Weights - Presumed Gestation Individual Data B-44 Table 16. Feed Consumption Values - Precohabitation Individual Data B-49 Table 17. Maternal Feed Consumption Values - Presumed Gestation - Individual Data B-54 Table 18. Caesarean-Sectioning Observations - Individual Data B-59 Table 19. Litter Observations (Caesarean-Delivered Embryos or Fetuses) - Individual Data B-63 Table 20. Embryonal Vital Status or Fetal Sex, Vital Status and Body Weight - Individual Data B-67 Table 21. Placental Weights - Individual Data B-73 APPENDIX C: - PROTOCOL AND AMENDMENT C-1 to C-34 APPENDIX D- DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY D-1 APPENDIX E - TEMPERATURE AND RELATIVE HUMIDITY REPORTS E-1 to E-3 APPENDIX F - HISTORICAL CONTROL DATA F-1 to F-13 vi 0 007S6 SUBJECT APPENDIX G - STATEMENT OF THE STUDY DIRECTOR APPENDIX H - QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT PAGE G-1 H-1 to H-5 VII C0 0 7 S 7 418-013:PAGE 1-1 TITLE: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-013 SPONSOR'S STUDY NUMBER: T-6295.12 I. SUMMARY AND CONCLUSION A. Methods* Eighty virgin female Crl:CDBR VAF/Plus (Sprague-Dawley) rats were assigned to five dosage groups (Groups I through V), 16 rats per dosage group. The rats were administered the test article, PFOS (FC-95), or the vehicle, 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water), orally (via gavage), once daily beginning 42 days prior to cohabitation through either day 14 or 20 of presumed gestation (DG 14 or DG 20). Dosages were 0 (Vehicle), 0.1, 0.4, 1.6 and 3.2 mg/kg/day. The dosage volume was 5 mL/kg. The rats were observed for viability at least twice each day of the study and for clinical signs of effects of the test article before and approximately one hour after dosage and on the day sacrificed. Body weights were recorded daily during the dosage period and at sacrifice. Feed consumption values were recorded weekly to cohabitation and daily during presumed gestation. Urine and fecal samples were collected one day prior to initiation of cohabitation to the following morning and DGs 6 to 7, 14 to 15 and 20 to 21. Samples were frozen and shipped to the Sponsor for analysis. Blood samples were collected on the day cohabitation was initiated and on DGs 7, 15 and 21. Blood was centrifuged and the serum was frozen and shipped to the Sponsor for analysis. Rats were sacrificed on DG 15 or 21, as described below, and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. A liver section and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal 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 AMENDMENT). 000788 418-013:PAGE I-2 regions of each dam were collected, frozen and shipped to the Sponsor for analysis. Eight randomly selected female rats, with a confirmed date of mating, per dosage group were Caesarean-sectioned on DG 15. The gravid uterus was excised and weighed. Rats were examined for number and distribution of corpora lutea, implantation sites, viable and nonviable embryos. A sample of the amniotic fluid of each viable embryo was collected and pooled by litter. Each viable embryo was removed from the uterus with the attached placenta and pooled by litter. These samples were frozen shipped to the Sponsor for analysis. All remaining female rats in each dosage group were Caesarean-sectioned on DG 21 or estimated DG 21. The gravid uterus was excised and weighed. Individual placental weights were recorded. Rats were examined for number and distribution of corpora lutea, implantation sites, live and dead fetuses and early and late resorptions. Each fetus was removed from the uterus, weighed and examined for sex and gross external alterations. Samples of the amniotic fluid were collected when possible, and pooled by litter. Placentae were collected and pooled by litter. Blood samples were collected from each fetus, pooled by litter and centrifuged. All samples were frozen and shipped to the Sponsor for analysis. Lung and liver samples were collected from three fetuses per litter from two to five litters per dosage group. Sections from the half of the lungs and one lobe of the liver were shipped to Pathology Associates International, Durham, North Carolina, for possible future evaluation. The other half of the lungs and the other lateral lobe of the liver were frozen and shipped to the Sponsor for analysis. The livers of the remaining fetuses, plus any remaining liver from the three selected fetuses, were collected, pooled by litter and frozen. The remaining carcass (and head) of each fetus was frozen and shipped to the Sponsor for analysis. B. Results No deaths or premature deliveries were attributed to PFOS treatment. Three rats (one each in the 0.1, 0.4 and 1.6 mg/kg/day dosage groups) were sacrificed because the eye was injured during orbital sinus bleeding. One rat in the vehicle control group did not have a confirmed mating date and delivered on day 66 of the study. Two rats in the 3.2 mg/kg/day dosage group delivered on DG 21. All other rats survived to scheduled sacrifice. 0007S9 418-013:PAGE I-3 All adverse clinical observations during the precohabitation and gestation periods were considered unrelated to the test article and no gross lesions were revealed by necropsy. Body weight gains for the entire precohabitation period (DSs 1 to 42) were 88.8%, 80.8%, 66.3% and 17.4% of the control group value in the 0.1, 0.4, 1.6 and 3.2 mg/kg/day dosage groups, respectively. Body weights were 98.0%, 96.3%, 93.6% and 85.3% of the control group value in these four respective dosage groups on DS 42. As the result of random selection for assignment to Caesarean-sectioning on either DG 15 or DG 21, groups assigned to Caesarean-sectioning on DG 15 weighed less on DG 0 than groups assigned to Caesarean-sectioning on DG 21. Reflecting the random selection and differences in average body weights of the two subsets, as well as the relatively small group sizes, body weight gains differed in the two subsets. In both subsets, body weight gains were reduced on DGs 0 to 7 in groups administered 0.4 mg/kg/day and higher dosages of PFOS, after which body weight changes did not demonstrate a clear dosage-dependent pattern. Absolute (g/day) and relative (g/kg/day) feed consumption values were reduced in the 0.4, 1.6 and 3.2 mg/kg/day dosage groups in each week of the precohabitation period. Reflecting these effects of the test article, absolute and relative feed consumption values were reduced for the entire precohabitation period (DSs 1 to 42) in the 0.4, 1.6 and 3.2 mg/kg/day dosage groups. Groups administered 0.4 mg/kg/day and higher dosages of the test article had reduced absolute and relative feed consumption values during the first week of the gestation period. Absolute feed consumption values continued to be reduced for the remainder of the dosage period in the 0.4 and 1.6 mg/kg/day dosage groups assigned to DG 15 Caesarean-sectioning and in the 3.2 mg/kg/day dosage group for rats assigned to either DG 15 or DG 21 Caesarean-sectioning. No Caesarean-sectioning or litter parameters were affected by dosages of the test article as high as 3.2 mg/kg/day administered to rats Caesarean-sectioned on DG 15. The litter averages for implantations, litter sizes and live fetuses were reduced in the 3.2 mg/kg/day dosage group Caesarean-sectioned on DG 21. Values were below the ranges observed historically at the Testing Facility. Fetal body weights were also reduced in the 3.2 mg/kg/day dosage group. No other Caesareansectioning or litter parameters were affected by dosages of the test article as 0C079O 418-013:PAGE I-4 high as 3.2 mg/kg/day administered to rats Caesarean-sectioned on DG 21. No fetal gross external alterations were observed. C. Conclusion The purpose of this study, as stated in the protocol, was to evaluate the pharmacokinetics of PFOS in Fo generation dams and F1 generation fetuses following PFOS treatment to female rats during premating and gestation. This reports includes study data from the in-life portion which was conducted at Argus Research Laboratories, Inc. The PK samples that were collected during the inlife portion of the study were sent to the Sponsor for analysis and will be reported separately. It is the responsibility of the Sponsor, 3M Corporate Toxicology, to combine the in-life results with the analytical results into a final pharmacokinetic report. The results for this study mimic those reported in the Combined Oral (Gavage) Fertility, Developmental and Perinatal/Postnatal Reproduction Toxicity Study of PFOS in Rats (Argus Research Laboratories, Inc., Protocol 418-008), reported 10 June 1999. Mildred S. Christian, Ph.D., Fellow, ATS Executive Director of Research Date Alan M. Hoberman, Ph.D., DABT Director of Research Date Associate Director of F^search and Study Director Date 0G0791 418-013:PAGE 11-1 II. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study: A.1. Sponsor: 3M Corporate Toxicology, 3M Center, Building 220-2E-02, St. Paul, Minnesota 55144-1000 A.2. Testina Facility: Argus Research Laboratories, Inc., 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 A.3. Study Number: 418-013 A.4. Sponsor's Study Number: T-6295.12 A.5. Purpose of the Study: The purpose of this study was to evaluate the pharmacokinetics of PFOS in Fo generation dams and F1 generation fetuses following PFOS treatment of Crl:CDBR VAF/Plus female rats during premating and gestation. A.6. Study Design: The requirements of the U.S. Food and Drug Administration (FDA)(1) were used as the basis of study design. A.7. Regulatory Compliance: The study was conducted in compliance with Good Laboratory Practice (GLP) regulations of the U.S. Food and Drug Administration (FDA)(2), the Japanese Ministry of Health and Welfare (MHW )<3) and the European Economic Community (EEC)<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. 000792 418-013:PAGE II-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: Marvin T. Case, D.V.M., Ph.D. A.10. Alternate Study Monitor: Andrew M. Seacat, Ph.D. A.11. Study Director: Raymond G. York, Ph.D., DABT (Associate Director of Research) A.12. Technical Performance: John F. Barnett, B.S. (Director of Laboratory Operations) Margaret M. Martin (Research Associate) Corrie L. Pabst (Quality Control Associate) A.13. Report Preparation: Raymond G. York, Ph.D., DABT Jo Ann Frazee, M.S. (Study Coordinator) Susan K. Bradshaw, B.S. (Data Management Specialist) Karen G. Parker, A.A. (Report Administrator) A.14. Report Review: Mildred S. Christian, Ph.D., Fellow, ATS (Executive Director of Research) Allan M. Hoberman, Ph.D, DABT (Director of Research) A.15. Date Protocol Signed: 9 November 1998 CO0793 418-013:PAGE II-3 A.16. Dates of Technical Performance: Rat Arrival Date Dosage Period (42 days prior to cohabitation until DGa 14 or 20) Cohabitation Period Caesarean-Sectioning Period (DG 15) Caesarean-Sectioning Period (DG 21) 10 NOV 98 16 NOV 9 8 -2 1 JAN 99 27 DEC 98 PM - 01 JAN 99 AM 12 JAN 9 9 - 13 JAN 99 18 JAN 9 9 - 2 2 JAN 99 A. 17. Records Maintained: The original report, raw data and reserve samples of the bulk test article and vehicle components are retained in the archives of Argus Research Laboratories, Inc. Any preserved tissues are retained in the archives of the Testing Facility for one year after the mailing of 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 will be returned to the Study Monitor upon completion of all work with the test article. B. Test Article Information: B.1. Description: PFOS (FC-95) - an off-white powder B.2. Lot/Batch Number: 217 (Expiration date: May 2000) B.3. Date Received and Storage Conditions: The test article was received on 21 October 1998, and stored at room temperature. B.4. Special Handling Instructions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles or safety glasses and a face-shield) were taken when handling the bulk test article and prepared suspensions. a. DG is used as an abbreviation for day of (presumed) gestation. 000794 418-013.PAGE II--4 B. 5. Analysis of Puritv: Information regarding the identity, composition, strength and purity of the test article is on file with the Sponsor. C. Vehicle Information: C.1. Description: 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). C.2. Lot Numbers: Tween 80 - M29477 and M03H05 C.3. Date Received and Storage Conditions: The Tween 80 was received from J.T. Baker, Phillipsburg, New Jersey, on 17 September 1998 (Lot Number M29477) and 3 December 1998 (Lot Number M03H05), and stored at room temperature. The R.O. deionized water is available from a continuous source at the Testing Facility and is maintained at room temperature. C.4. Special Handling Instructions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles or safety glasses and a face-shield) were taken when handling the vehicle components and prepared vehicle. C. 5. Analysis of Puritv: Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. D. Test Article Preparation and Storage Conditions: Suspensions of PFOS were prepared daily at concentrations of 0, 0.02, 0.08, 0.32 and 0.64 mg/mL. Prepared formulations were stored at room temperature. 000795 D.1. Sample Information: 418-013:PAGE II-5 Date Storage/Shipping Date Sample Type Size Retained Conditions Shipped To Shipped Concentration 2mL* 15 NOV 98 Frozen Sponsor 16 NOV 98 (all levels) 20 JAN 99 20 JAN 99 Bulk Test Article Testing Facility Reserve 1 g 12 NOV 98 Room temperature Archives 25 NOV 98 Vehicle Components Reserve Tween 80 5 mL 12 NOV 98b Room temperature Testing Facility 25 NOV 98 10 DEC 98c Archives 27 JAN 99 R.O. deionized water 5 mL 12 NOV 98 Room temperature Testing Facility 25 NOV 98 Archives a. Duplicate samples were taken from the first and last preparation on the day prepared. One sample of each set was shipped for analysis. The remaining samples were retained at the Testing Facility as backups. Backup samples are stored frozen (-70C or below) and will be discarded at the request of the Sponsor. b. Lot Number M29477. c. Lot Number M03H05. D.2. Analytical Results: Data verifying the stability of the test article in the vehicle for 48 hours under the conditions of administration and the stability of the bulk test article are on file with the Sponsor. Homogeneity of prepared formulations is on file with the Sponsor. Results of the concentration analysis were not available at the time of the writing of this report. E. Test System : E.1. Species: Rat E.2. Strain: Crl:CDBR VAF/Plus (Sprague-Dawley) E.3. Supplier (Source): Charles River Laboratories, Inc., Raleigh, North Carolina E.4. Sex: Female (Note: Male rats were used only for the purposes of breeding and are not considered part of the Test System.) 000796 418-013:PAGE li-6 E.5. Rationale for Test System: The Crl:CDBR VAF/Plus (Sprague-Dawley) rat was selected as the Test System because: 1) this strain of rat was used in the reproductive and developmental toxicity studies; 2) historical data and experience exist at the Testing Facility'5'75; and 3) the test article is pharmacologically active in the species and strain. E.6. Test System Data: Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) on the Day After Arrival Weight (g) at Study Assignment 122 07 SEP 98 65 days 193-223 208 - 224 E.7. Breeder Male Rat Data: Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) on the Day After Arrival Weight (g) at Cohabitation 112 13 JAN 98 78 days 300 - 356 558 - 871 E.8. Method of Randomization: Upon arrival, rats were assigned to individual housing on the basis of computer generated random units. After acclimation, virgin female rats were selected for study on the basis of physical appearance and body weight recorded during acclimation. Female rats were assigned to five dosage groups (Groups I through V), 16 rats per dosage group, using a computer-generated (weight-ordered) randomization procedure. Within each dosage group, consecutive order was used to assign female rats to cohabitation with breeder male rats, one male rat per female rat. A table of random units was used to select eight female rats with confirmed mating dates per dosage group for Caesarean-section examinations on DG 15. The remaining female rats in each dosage group were Caesareansectioned on DG 21a. a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), item 1. 000797 E. 9. System of Identification: 418-013:PAGE II-7 Male rats were given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats were assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study. Each rat was individually identified with a Monel self-piercing ear tag (Gey Band and Tag Co., Inc., No. MSPT 20101) inscribed with the rat's designated unique permanent number. F. Husbandry: F.1. Research Facility Registration: USDA Registration No. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 et seq. F.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%. See APPENDIX E (TEMPERATURE AND RELATIVE HUMIDITY REPORTS.) F.3. Housing: All cage sizes and housing conditions were in compliance with the Guide for the Care and Use o f Laboratory Anim als(8). Female rats were individually housed in stainless steel, wire-bottomed cages except during the cohabitation period and during collection intervals for urine and fecal samples. During cohabitation, each pair of rats was housed in the male rat's cage. During collection intervals for urine and fecal samples the female rats were individually housed in metabolism cages. No nesting materials were supplied because the female rats were sacrificed before parturition was expected. F.4. Lighting: An automatically-controlled fluorescent light cycle was maintained at 12-hours light: 12-hours dark, with each dark period beginning at 1900 hours EST. 000798 418-013:PAGE II-8 F.5. Sanitization: Cage pan liners were changed approximately three times each week. Cages were changed approximately every other week. F.6. Feed: Rats were given ad libitum access to Certified Rodent Diet #5002 (PMI Nutrition International, St. Louis, Missouri) in individual feeders. F.7. Feed Analysis: Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration limits 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 Study Director nor the Sponsor was aware of any agent present in the feed that was known to interfere with the results of this study. F.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 individual water bottles and/or from an automatic watering access system. Chlorine was added to the processed water as a bacteriostat. F.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 Study Director nor the Sponsor was aware of any agent present in the water that was known to interfere with the results of this study. 000799 G. Methods: 418-013:PAGE II-9 G.1. Dosage Administration: Dosage Group I II III IV V Dosage (mg/kg/day)a 0 (Vehicle) Concentration (mg/mL) 0 Dosage Volume (mL/kg) 5 0.1 0.02 5 0.4 0.08 5 1.6 0.32 5 3.2 0.64 5 Number of Female Rats 16 16 16 16 16 Assigned Rat Numbers 19501 - 19516 19517- 19532 19533 - 19548 19549- 19564 19565 - 19580 a. The test article was considered 100% pure for the purpose of dosage calculations. G.2. Rationale for Dosage Selection: Dosages were selected on the basis of a previous study conducted with the test article (Argus Research Laboratories, Inc., Protocol 418-008). G.3. Route of Administration: Oral (gavage) G.4. Rationale for Route of Administration: The oral (gavage) route was selected for use because: 1) this was the route of administration in the developmental and reproductive toxicology studies; and 2) it is one of the possible routes of human exposure. G.5. Freouencv of Administration: Appropriate dosages of the test article or vehicle were administered orally (via gavage) once daily to female rats beginning 42 days prior to cohabitation through either DG 14 or DG 20. The dosage volume was 5 mL/kg, adjusted daily on the basis of the individual body weights recorded before intubation. The rats were intubated once daily at approximately the same time each day. G.6. Length of Study: Approximately 11 weeks CQOSOO 418-013:PAGE 11-10 G.7. Method of Study Performance: The female rats were observed for viability at least twice each day of the study and for general appearance at least once during acclimation. The rats were also examined for clinical observations of effects of the test article, abortions, premature deliveries and deaths before and approximately one hour after dosage and once prior to sacrifice. Body weights were recorded at least once during acclimation, daily during the dosage period and at sacrifice. Feed consumption values were recorded at least once during the acclimation period, weekly to cohabitation and daily during presumed gestation. After 42 days of test article administration, 80 healthy virgin female rats were placed into cohabitation with 80 breeder male rats (one male rat per female rat in the male rat's cage)3. Mating was evaluated daily during the cohabitation period and confirmed by the observation of spermatozoa in a smear of the vaginal contents and/or a copuiatory plug in situ. Female rats considered to be at DG 0 were returned to individual housing. Urine and fecal samples were collected from female rats at the following intervals: one day prior to initiation of cohabitation to the following morning and from each of the mated female rats on DGs 6 to 7, 14 to 15 and 20 to 21. Following each 24-hour collection interval, samples were collected into centrifuge tubes, placed on dry ice and maintained frozen (-70C or below) until shipment (on dry ice) to the Sponsor for analysis. Blood samples were collected from each of the female rats following removal from metabolism caging (prior to test article or vehicle administration) on the day cohabitation was initiated (prior to cohabitation) and from each of the mated female rats on DGs 7, 15 and 21. On all days of collection except DGs 15 and 21 (dams at their terminal collection interval), blood samples (approximately 1 mL each) were collected from the orbital sinus. On DG 21, blood samples (approximately 4 mL each) were collected via the inferior vena cava. When blood collection on DG 15 was a final bleed for the respective dam (i.e., the dam was Caesarean-sectioned that day), the sample was approximately 4 mL and was collected via the inferior vena cava. Blood was transferred into serum separator tubes and spun in a refrigerated centrifuge. The serum was transferred into polypropylene tubes labeled with the study number, rat identification, date of collection, study day and collection timepoint. All samples were immediately frozen on dry ice and stored (-70C or below) until shipment (on dry ice) to the Sponsor for analysis. a. See APPENDIX D, item 1. 000801 418-013:PAGE 11-11 G.8. Gross Necropsy*: Rats were sacrificed by carbon dioxide asphyxiation on DG 15 or 21, as described below. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. A liver section (right lateral lobe) and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal regions (left side only) of each dam (except rats without confirmed dates of mating) were collected, frozen and stored (-70C or below) until shipment (on dry ice) to the Sponsor for analysis. To confirm the pregnancy status, uteri from rats that appeared nonpregnant were stained with 10% ammonium sulfide<9). Tissues with gross lesions were preserved in neutral buffered 10% formalin for possible future evaluation; all other maternal tissues were discarded. Representative photographs of maternal lesions are available in the raw data. G.8.a. DG 15 Caesarean-Sectioning Eight randomly selected female rats, with a confirmed date of mating, per dosage group were Caesarean-sectioned on DG 15. The gravid uterus was excised and weighed. Rats were examined for number and distribution of corpora lutea, implantation sites, viable and nonviable embryos. A viable embryo was defined as oval or crescent shaped, pink, firm and enclosed in an amniotic sac filled with clear fluid. A nonviable embryo is amorphous, small, pale pink to tan or deep red to black, soft and enclosed in an amniotic sac filled with clear, cloudy or opaque fluid. A sample of the amniotic fluid of each viable embryo was collected and pooled per litter, frozen and stored (-70C or below) until shipment, on dry ice, to the Sponsor for analysis. Each viable embryo was removed from the uterus with the attached placenta, pooled per litter frozen and stored (-70C or below) until shipment, on dry ice, to the Sponsor for analysis. G.8.b. DG 21 Caesarean-Sectioning All remaining female rats in each dosage group were Caesarean-sectioned on DG 21 (rats with confirmed dates of mating) or estimated DG 21 (rats without confirmed dates of mating). The gravid uterus was excised and weighed. Individual placental weights were recorded. Rats were examined for number and distribution of corpora lutea, 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 a. A table of random units was used to select one control group rat from which all tissues examined at necropsy were retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions. 000502 418-013:PAGE 11-12 which the occurrence of organogenesis was grossly evident. A live fetus was defined as a term fetus that responded to stimuli. Nonresponding term fetuses are considered to be dead (there were no dead fetuses). Dead fetuses and late resorptions are differentiated by the degree of autolysis present; marked to extreme autolysis indicated that the fetus was a late resorption. Each fetus was removed from the uterus, placed in an individual container and identified with a tag noting the study number, litter number, and uterine distribution. Each fetus was subsequently weighed and examined for sex and gross external alterations. Live fetuses were sacrificed via decapitation. Samples of the amniotic fluid (when possible) and the placenta of each fetus were collected, individually pooled per litter, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Blood samples were collected from each fetus via decapitation, pooled per litter and transferred into serum separator tubes. The samples were spun in a refrigerated centrifuge. The serum was transferred into polypropylene tubes labeled with the study number, litter identification, date of collection, study day and collection timepoint. All samples were immediately frozen on dry ice and stored (-70C or below) until shipment (on dry ice) to the Sponsor for analysis. Lung and liver samples were collected from three fetuses per litter from two to five litters per dosage group. One half of the lungs and one lateral lobe of the liver were individually retained in neutral buffered 10% formalin in scintillation vials. Prior to fixation, several (minimum of two) sections (approximately 1 mm thick) of this half of the lungs and this lobe of the liver were removed using a scalpel and retained in McDowell-Trump's Fixative (stored under refrigeration in scintillation vials). The lung and liver sections in McDowell-Trump's Fixative and the lung and liver samples in neutral buffered 10% formalin were shipped (on cold packs), for possible future evaluation by electron microscopy or light microscopy, respectively, to Pathology Associates International, Durham, North Carolina. The other half of the lungs and the other lateral lobe of the liver were flash frozen in liquid nitrogen and stored in heat-sealable pouches on dry ice. These flash frozen lung and liver samples were maintained frozen (-70C or below) until shipment (on dry ice) to the Sponsor for analysis. The livers of the remaining fetuses, plus any remaining liver from the three selected fetuses, were collected, pooled by litter and frozen. The remaining carcass (and head) of each fetus was frozen. These samples were maintained frozen (-70C or below) until shipment (on dry ice) to the Sponsor for analysis. G.8.C. Moribund Sacrifice or Premature Delivery Rats that were sacrificed because of moribund condition or premature delivery were examined for the cause on the day the observation was made. The rats 000503 418-013:PAGE 11-13 were examined for gross lesions. A liver section (right lateral lobe) and the milksecreting glands from the axillary, thoracic, abdominal and inguinal regions of each dam (left side only) were collected, frozen and stored (-70C or below) until shipment (on dry ice) to the Sponsor for analysis. Pregnancy status and uterine contents were recorded. Delivered pups were examined to the extent possible, using the same methods described for term fetuses. Uteri of apparently nonpregnant rats were stained with 10% ammonium sulfide to confirm the absence of implantation sites(9). G.9. Statistical Analyses: Averages and percentages were calculated. Litter values were used where appropriate. 0G0304 III. RESULTS 418-013:PAGE 111-1 A. Mortality. Premature Delivery, Clinical and Necropsy Observations (Summary - Table 1: Individual Data - Tables 12 and 13) A.1. Mortality and Premature Delivery No deaths or premature deliveries were attributed to PFOS treatment. Three rats (one each in the 0.1, 0.4 and 1.6 mg/kg/day dosage groups) were sacrificed because the eye was injured during orbital sinus bleeding. One rat in the vehicle control group did not have a confirmed mating date and delivered on day 66 of the study. Two rats in the 3.2 mg/kg/day dosage group delivered on DG 21. All other rats survived to scheduled sacrifice. A.2. Clinical Observations All adverse clinical observations during the precohabitation and gestation periods were considered unrelated to the test article because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in only one or two rats. These observations included missing or swollen forepaw digit, scabs on forepaw, swollen forepaw, localized alopecia on the limbs, head, back and/or underside, chromorhinorrhea, chromodacryorrhea, swollen ears, dental problems (missing, broken and/or misaligned incisors), swollen snout, corneal hemorrhage, corneal opacity, axillary mass, exophthalmos, enophthalmos, lenticular opacity, lacrimation, ungroomed coat, traumatized cornea, cloudy, white film on eye, missing right eye, large eye, dark red eye, and lids unable to close. A. 3. Necropsy Observations No gross lesions were revealed by necropsy. B. Body Weights. Uterine Weights and Body Weight Changes (Figures 1 through 3: Summaries - Tables 2 through 5: Individual Data - Tables 14 and 15) B.1. Precohabitation Body weight gains for the entire precohabitation period (DSs 1 to 42) were 88.8%, 80.8%, 66.3% and 17.4% of the control group value in the 0 .1 ,0 .4 , 1.6 and 3.2 mg/kg/day dosage groups, respectively. Body weights were 98.0%, 96.3%, 93.6% and 85.3 % of the control group value in these four respective dosage groups on DS 42. 000305 B. 2. Gestation 418-013:PAGE III-2 As the result of random selection for assignment to Caesarean-sectioning on either DG 15 or DG 21, groups assigned to Caesarean-sectioning on DG 15 weighed less on DG 0 than groups assigned to Caesarean-sectioning on DG 21. Reflecting the random selection and differences in average body weights of the two subsets, as well as the relatively small group sizes, body weight gains differed in the two subsets. In both subsets, body weight gains were reduced on DGs 0 to 7 in groups administered 0.4 mg/kg/day and higher dosages of PFOS, after which body weight changes did not demonstrate a clear dosage-dependent pattern. Gestation body weights and body weight gains were unaffected by the 0.1 mg/kg/day dosage of the test article. Gravid uterine weights were unaffected by dosages of the test article as high as 3.2 mg/kg/day. C. Absolute (q/dav) and Relative (q/kq/dav) Feed Consumption Values (Summaries - Tables 6 through 9: Individual Data - Tables 16 and 17) C.1. Precohabitation Absolute (g/day) and relative (g/kg/day) feed consumption values were reduced in the 0.4, 1.6 and 3.2 mg/kg/day dosage groups in each week of the precohabitation period. Reflecting these effects of the test article, absolute feed consumption values were 94.8%, 92.2% and 83.8% of the control group value, and relative feed consumption values were 95.1%, 94.3% and 90.7% of the control group value for the entire precohabitation period (DSs 1 to 42) in the 0.4, 1.6 and 3.2 mg/kg/day dosage groups. Absolute and relative feed consumption values during the precohabitation period were unaffected by the 0.1 mg/kg/day dosage of the test article. C.2. Gestation Groups administered 0.4 mg/kg/day and higher dosages of the test article had reduced absolute and relative feed consumption values during the first week of the gestation period (no values were available for evaluation for the 1.6 mg/kg/day dosage group assigned to Caesarean-sectioning on DG 15). Absolute feed consumption values continued to be reduced for the remainder of the dosage period in the 0.4 and 1.6 mg/kg/day dosage groups assigned to DG 15 Caesarean-sectioning and in the 3.2 mg/kg/day dosage group for rats assigned to either DG 15 or DG 21 Caesarean-sectioning. Relative feed consumption values tended to be increased over the control group values for the 1.6 and 3.2 mg/kg/day dosage groups after DG 10. 0C0806 418-013:PAGE III-3 Absolute and relative feed consumption values during gestation were unaffected by the 0.1 mg/kg/day dosage of the test article. D. Caesarean-Sectioning. Litter and Fetal Gross External Observations (Summaries-Tables 10 and 11: Individual Data - Tables 18 through m D.1. Day 15 of Gestation Pregnancy occurred in 6 (75.0%), 7 (87.5%), 8 (100.0%), 6 (85.7%) and 8 (100.0%) of the rats in each dosage group. No Caesarean-sectioning or litter parameters were affected by dosages of the test article as high as 3.2 mg/kg/day administered to rats Caesarean-sectioned on DG 15. The litter averages for corpora ltea, implantations, viable and nonviable embryos and percent nonviable embryos per litter were comparable among the five dosage groups. No dam had a litter consisting of only nonviable embryos. D.2. Dav 21 of Gestation Pregnancy occurred in 7 (100.0%), 7 (100.0%), 5 (83.3%), 2 (50.0% ) and 6 (100.0%) rats with a confirmed mating date in each dosage group. One rat in each of the 0.1 and 0.4 mg/kg/day dosage groups was moribund sacrificed on DG 0 and the pregnancy status could not be determined and two rats in the 3.2 mg/kg/day dosage group delivered before Caesarean-sectioning on DG 21, as previously described. As a result, Caesarean-sectioning observations were based on 7, 7, 5, 2 and 4 pregnant rats with one or more live fetuses in Groups I through V, respectively. The litter averages for implantations, litter sizes and live fetuses were reduced in the 3.2 mg/kg/day dosage group. Values were below the ranges observed historically at the Testing Facility3. Fetal body weights were also reduced in the 3.2 mg/kg/day dosage group. No other Caesarean-sectioning or litter parameters were affected by dosages of the test article as high as 3.2 mg/kg/day administered to rats Caesarean-sectioned on DG 21. The litter averages for corpora ltea, early and late resorptions, placental weights, percent live male fetuses and percent resorbed conceptuses were comparable among the five dosage groups. No dam had a litter consisting of only resorbed conceptuses, and there were no dead fetuses. No fetal alterations were identified at gross external examination. a. See APPENDIX F (HISTORICAL CONTROL DATA). GO0807 REFERENCES 418-013:PAGE III-4 1. 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 Standard for 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 o f an OECD decision/recommendation on compliance with principles o f good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. 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 Cri:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 8. Institute of Laboratory Animal Resources (1996). Guide fo r the Care and Use o f Laboratory Animals. National Academy Press, Washington, D.C. 9. Salewski, E. (1964). Farbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. 0C0S08 APPENDIX A REPORT FIGURES 000S09 300 290 I BODY WEIGHTS PRECOHABITATION Figure 1 0 (VEHICLE) MG/KG/DAY 0.1 MG/KG/DAY n 0.4MG/KG/DAY 16 MG/KG/DAY X 3 2MG/KG/DAY TSQGO 418-013:PAGE A-1 220 a. Lastvaluerecorded beforecohabitation 15 22 29 DAY OF STUDY I 36 Aia MATERNAL BODY WEIGHTS RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION Figure 2 360 I I 340 0 (VEHICLE) MG/KG/DAY 320 CD 300 I- X CD 1280 260 r_r 0.1 MG/KG/DAY -n - 0.4MG/KG/DAY -A1.6MG/KG/DAY X 3 2 MG/KG/DAY x240 418-013: PAGE A-2 000811 220 6 7 9 10 11 12 13 14 15 DAY OF GESTATION MATERNAL BODY WEIGHTS RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION Figure 3 450 0 (VEHICLE) MG/KG/DAY 01 MG/KG/DAY 0.4MG/KG/DAY - 16 MG/KG/DAY X 3.2MG/KG/DAY 200 6 7 8 10 11 12 13 14 15 18 17 18 19 20 21 DAY OF GESTATION 418-013: PAGE A-3 COOS12 APPENDIX B REPORT TABLES 000813 418-013: PAGE B-1 000814 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 1 (PAGE 1): CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY (See footnotes on the last page of this table.) DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 0.1 MORTALITY DELIVERED AND SACRIFICED MORIBUND SACRIFICED 11 la 0 0 Id PRECOHABITATION (DAY 1 OF STUDY TO THE DAY OF COHABITATION) MAXIMUM POSSIBLE INCIDENCE 672/ 16 672/ 16 FOREPAW: THIRD DIGIT MISSING 0/ 0 0/ 0 LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE 40/ 1 40/ 1 0/ 0 43/ 2 43/ 2 0/ 0 FOREPAW: THIRD DIGIT SWOLLEN 0/ 0 0/ 0 FOREPAWS: SCAB(S) 0/ 0 0/ 0 CHROMORHINORRHEA 0/ 0 0/ 0 CHROMODACRYORRHEA 0/ 0 5/ 1 EARS: SWOLLEN 0/ 0 0/ 0 FOREPAWS: SWOLLEN 0/ 0 0/ 0 INCISORS: TOTAL MISSING/BROKEN MISALIGNED 0/ 0 0/ 0 0/ 0 21/ 1 12/ 1 9/ 1 SNOUT: SWOLLEN 0/ 0 1/ 1 MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. III 0.4 1 0 le 672/ 16 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 IV 1.6 1 0 If 672/ 16 0/ 0 26/ 2 14/ 1 12/ 1 0/ 0 14/ 1 5/ 4 0/ 0 3/ 1 2/ 1 0/ 0 0/ 0 0/ 0 0/ 0 V 3.2 2 2b ,c 0 672/ 16 34/ 1 9/ 1 9/ 1 0/ 0 3/ 1 3/ 1 1/ 1 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 PROTOCOL 416-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295,12) TABLE 1 (PAGE 2): CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY (See footnotes on the last page of this table.) DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 0.1 PRESUMED GESTATION: q MAXIMUM POSSIBLE INCIDENCE 282/ 15 283/ 16 LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE HEAD BACK 3/ 1 3/ 1 0/ 0 0/ 0 0/ 0 37/ 3 23/ 2d 0/ 0 14/ 1 0/ 0 FOREPAW: THIRD DIGIT MISSING 0/ 0 0/ 0 CORNEAL HEMORRHAGE 0/ 0 0/ 0 CORNEAL OPACITY 0/ 0 27/ 3d RIGHT AXILLA: MASS 0/ 0 0/ 0 EXOPHTHALMOS 0/ 0 4/ 2d CHROMODACRYORRHEA 11/ 1 1/ Id FOREPAWS: SCAB(S) 0/ 0 0/ 0 ENOPHTHALMOS 0/ 0 0/ 0 LENTICULAR OPACITY 16/ la 0/ 0 LACRIMATION 0/ 0 0/ 0 RIGHT EYE: LARGE 0/ 0 1/ Id RIGHT EYE: DARK RED 0/ 0 1/ Id MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. o III 0.4 261/ 15 3/ 1 0/ 0 3/ 1 0/ 0 0/ 0 0/ 0 0/ 0 20/ 2e 0/ 0 4/ 2e 1/ le 0/ 0 8/ 1 3/ 1 2/ 1 1/ le 1/ le IV 1.6 216/ 12 45/ 3 37/ 2 16/ 1 0/ 0 7/ 1 0/ 0 6/ 1 0/ 0 0/ 0 1/ 1 0/ 0 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 V 3.2 260/ 14 46/ 4 43/ 3c 3/ i 0/ 0 0/ 0 22/ 1 16/ 1 16/ 1 11/ 1 2/ 1 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 418-013: PAGE B-2 PROTOCOL 418-013; ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR 'S STUDY NUMBER: T- 6295.12) TABLE 1 (PAGE 3): CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV V 0 (VEHICLE) 0.1 0.4 1.6 3.2 PRESUMED GESTATION: q MAXIMUM POSSIBLE INCIDENCE 282/ 15 283/ 16 261/ 15 216/ 12 260/ 14 RIGHT EYE: LIDS UNABLE TO CLOSE 0/ 0 1/ Id 1/ le 0/ 0 0/ 0 CHROMORHINORRHEA SNOUT: SWOLLEN UNGROOMED COAT 7/ 2 0/ 0 0/ 0 0/ 0 3/ 2d 0/ 0 1/ 1 1/ le 1/ i 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 TRAUMATIZED CORNEA 0/ 0 33/ 3 0/ 0 0/ 0 0/ 0 RIGHT EYE: CLOUDY, WHITE FILM 0/ 0 11/ 1 0/ 0 0/ 0 0/ 0 RIGHT EYE: MISSING 0/ 0 11/ 1 0/ 0 0/ 0 0/ 0 INCISORS: MISSING/BROKEN 0/ 0 7/ 1 0/ 0 0/ 0 0/ 0 PERSISTENT ADVERSE CLINICAL OBSERVATIONS WERE CONFIRMED AT NECROPSY, NO ADDITIONAL GROSS LESIONS WERE IDENTIFIED 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 19503 did not have a confirmed mating date and delivered on day 66 of study. b. Dam 19571 delivered on day 21 of gestation. c. Dam 19579 delivered on day 21 of gestation. d. Rat 19520 was moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. e. Rat 19541 was moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. f. Rat 19557 was moribund sacrificed on day 43 of study. g. Restricted to rats with a confirmed mating date. 418-013: PAGE B-3 9TS000 418-013-.PAGE B-4 C00S17 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 2 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) RATS TESTED N 16 BODY WEIGHT (G) DAY 1 MEAN+S.D 227.3 + 7.2 DAY 8 MEAN+S.D 243.4 10.2 DAY 15 MEAN+S.D 252.9 + 11.6 DAY 22 MEAN+S.D 266.3 + 12.8 DAY 29 MEAN+S.D 270.9 + 16.0 DAY 36 MEAN+S.D 273.3 + 18.6 DAY 42a MEAN+S.D 278.9 + 18.2 DAY = DAY OF STUDY a. Last value recorded before cohabitation. II 0.1 16 227.4 + 4.3 244.6 + 6.8 252.1 + 8.6 263.9 + 10.5 266.9 + 11.5 272.2 12.7 273.2 + 15.4 hi 0.4 16 226.9 + 6.2 242.0 + 9.4 249.0 + 11.7 261.1 + 15.1 264.0 + 17.1 270.3 + 17.9 268.6 + 23.4 IV 1.6 16 226.9 + 6.9 241.7 + 10.1 249.8 11.2 258.7 + 12.3 258.4 + 14.4 263.0 + 15.0 261.1 + 15.9 V 3.2 16 228.8 + 7.0 240.6 + 10.3 242.9 + 10.7 246.6 + 12.4 243.9 + 13.7 242.2 + 15.1 237.8 + 14.2 418-013.PAGE B-5 000318 PROTOCOL 418-011: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 3 (PAGE 1): BODY HEIGHT CHANGES - PRECOHABITATION - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) RATS TESTED N 16 BODY HEIGHT CHANGE (G) DAYS 1 - 8 MEAN+S.D . +16.1 + 5.6 DAYS 8 - IS MEAN+S.D . +9.5 + 5.6 DAYS 15 - 22 MEAN+S.D . 13.4 + 5.8 DAYS 22 - 29 MEAN+S.D. +4.6 + 6.6 DAYS 29 - 36 MEAN+S.D . +2.4 7.1 DAYS 36 - 42a MEAN+S.D . +5.6 + 5.2 DAYS 1 - 42a MEAN+S.D. 51.6 + 16.9 DAYS = DAYS OF STUDY a. Last value recorded before cohabitation. II 0.1 16 +17.2 + 5.1 +7.5 + 4.7 +11.8 + 4.8 +3.0 + 5.7 +5.3 + 5.4 +0.9 + 6.1 45.8 14.6 III 0.4 16 +15.1 + 5.1 +7.0 + 4.4 +12.1 + 5.2 +2.9 + 6.1 +6.3 6.4 -1.7 + 23.6 +41.7 + 22.6 IV 1.6 16 +14.8 + 5.7 +8.1 + 4.7 +8.9 + 4.2 -0.2 + 4.8 +4.6 + 4.4 -1.9 + 6.0 34.2 + 13.2 V 3.2 16 11.9 6.2 +2.3 + 4.6 +3.6 + 4.9 -2.6 + 6.0 -1.7 + 6.4 -4.5 + 5.7 +9.0 + 11.3 418-013: PAGE B-6 000319 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 1): MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV 0 (VEHICLE) 0.1 0.4 1.6 RATS TESTED N 8 8 8 8 INCLUDED IN ANALYSES N a 8 8 7a PREGNANT N6 7 8 6 MATERNAL BODY WEIGHT (G) DAY 0 MEAN+S.D . 281.0 4 28.9 276.0 4 10.5 270.4 4 14.4 267.2 4 16.4 DAY 1 MEAN+S.D . 287.0 4 26.0 282.4 4 13.0 275.1 4_ 20.8 267.2 4 15.1 DAY 2 MEAN+S.D. 294.3 4 25.1 290.0 4 11.2 282.8 4 19.3 275.3 4 17.0 DAY 3 DAY 4 MEANS .D . MEAN+S.D . 297.2 4 27.5 302.8 + 28.4 289.1 4_ 11.2 293.6 _4 10.5 285.0 16.9 286.8 4 17.4 274.8 4 17.4 275.8 4 15.8 DAY 5 MEAN+S.D . 304.2 4 26.2 296.6 4 12.4 289.4 4 16.3 276.8 4 19.0 DAY 6 MEAN+S.D . 308.7 4 27.1 302.0 +_ 14.2 291.9 4 17.3 278.3 4 16.3 DAY 7 MEAN+S.D . 310.2 4 28.2 302.6 4 12.2 285.5 4 20.6 273.0 4 14.5 DAY 8 MEAN+S.D . 309.2 4 25.0 302.8 4 12.0 295.8 4 21.3 279.0 4 17.7 DAY 9 MEAN+S.D . 315.7 23.1 308.0 4 13.7 294.4 4 17.9 280.3 4 16.1 DAY 10 MEAN+S.D . 319.7 25.0 313.7 17.9 296.9 4 19.0 285.3 4 15.6 DAY 11 MEAN+S.D. 327.3 4 24.5 314.6 4 13.9 302.4 4 18.6 292.2 4 16.8 DAY 12 MEAN+S.D. 330.5 23.9 321.7 4 15.0 309.1 4 18.7 297.2 4 18.9 DAY = DAY OF GESTATION a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. to < 3. 8 8 8 240.5 4 10.5 238.9 4 13.6 240.2 4_ 13.4 240.0 4 14.8 242.1 4 10.8 245.6 4_ 12.2 248.4 4 11.7 243.4 4 13.0 246.5 4 9.7 252.2 4 10.2 262.0 4 10.5 265.9 4 9.9 269.1 4 9.5 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 2) : MATERNAL BODY HEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV 0 (VEHICLE) 0.1 0.4 1.6 RATS TESTED N 8 8 8 8 INCLUDED IN ANALYSES N 8 a 8 7a PREGNANT N6 7 8 6 MATERNAL BODY WEIGHT (G) DAY 13 MEAN+S.D . 335.3 +24.6 324.3 + 15.6 314.0 + 19.8 298.8 + 17.8 DAY 14 MEAN+S.D. 339.2 + 25.8 328.4 + 15.5 317.8 + 19.2 305.5 + 20.8 DAY 15 MEAN+S.D . 345.2 27.8 331.1 + 18.6 319.9 + 22.4 309.8 + 17.7 GRAVID UTERINE WEIGHTS (G) MEAN+S.D . 19.8 4.3 17.5 + 5.2 16.3 5.6 17.6 + 1.3 DAY 15C b MEAN+S.D. 325.3 + 25.7 313.7 + 15.7 303.6 + 19.5 292.2 + 16.9 DAY = DAY OF GESTATION a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. b. 15C = Corrected maternal body weight (day 15 of gestation body weight minus the gravid uterine weight). V 3 .2 8 8 8 276.6 13.3 281.6 + 12.6 280.4 13.3 17.1 + 4.0 263.3 11.3 418-013: PAGE B-7 0G Q 82 o PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 3) : MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV V 0 (VEHICLE) 0.1 0.4 1.6 3. 2 RATS TESTED N 8 8 8 88 INCLUDED IN ANALYSES N 7a 7b 6a ,b 4a 4C 6a PREGNANT N7 7 5 2 6 MATERNAL BODY WEIGHT (G) DAY 0 MEAN+S.D . 281.6 4 14.5 282.3 4 19.1 292.8 4 22.7 273.0 4_ 1.4 242.0 4_ 14.7 DAY 1 MEAN+S.D. 290.3 + 15.6 290.4 4 17.9 294.4 4 24.7 277.5 4 0.7 242.8 4_ 17.3 DAY 2 MEAN+S.D. 293.8 4 14.8 298.3 4 16.5 301.2 4 24.8 282.5 4 0.7 242.7 4 18.2 DAY 3 MEAN+S.D . 298.4 4 12.6 300.4 4 19.3 302.6 4 23.3 284.5 4 2.1 243.2 4_ 20.3 DAY 4 MEAN+S.D. 30Q.4 4 13.4 304.0 4 14.2 305.4 _4 24.5 283.5 4 0.7 241.8 4 21.3 DAY 5 MEAN+S.D. 304.8 4 9.6 307.4 4 16.8 307.8 4 26.1 289.5 4 2.1 244.8 4_ 22.8 DAY 6 MEAN+S.D . 307.6 4 10.7 310.6 4 17.4 310.6 4 26.6 290.0 4 2.8 245.2 24.2 DAY 7 MEAN+S.D . 301.0 4 12.8 313.3 4 18.3 307.4 4 25.9 285.5 4 0.7 242.0 _4 19.8 DAY 8 MEAN+S.D . 305.8 4 12.8 314.4 4 16.6 309.0 4 26.1 286.0 4 1.4 243.5 4 21.0 DAY 9 MEAN+S.D. 311.0 4 10.4 318.7 4 15.4 313.4 4 27.3 292.5 4 2.1 249.0 4 20.5 DAY 10 MEAN+S.D . 317.0 4 8.8 323.1 4 16.9 316.6 4 26.3 295.0 4 7.1 256.3 4 18.1 DAY 11 MEAN+S.D . 321.7 4 10.1 329.6 4 15.2 320.2 4 23.7 300.5 4 7.8 260.7 4 21.2 DAY 12 MEAN+S.D. 326.1 4 11.6 330.8 4 16.8 327.2 4 28.2 304.5 4 12.0 263.5 4 21.0 DAY DAY OF GESTATION a. Excludes values for rats that did not have a confirmed mating date. b. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. c. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). 418-013:PAGE B-8 00082 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 4): MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV V 0 (VEHICLE) 0.1 0.4 1.6 3 .2 RATS TESTED N 8 8 a 8 8 INCLUDED IN ANALYSES N 7a 7b 6a ,b 4a c 6a PREGNANT N7 7 5 2 6 MATERNAL BODY WEIGHT (G) DAY 13 DAY 14 MEAN+S.D . MEAN+S.D . 329.1 10.1 334.0 + 12.1 336.0 i 14.9 340.3 + 17.4 331.8 + 28.0 337.0 + 28.7 312.5 4.9 317.0 + 12.7 26B .2 21.2 272.2 f 22.2 DAY 15 MEAN+S.D . 336.0 16.0 339.7 17.5 329.2 + 24.3 323.5 6.4 273.2 f 20.3 DAY 16 MEAN+S.D . 345.7 13.8 350.7 + 20.9 349.4 + 29.5 339.0 15.6 283.8 20.8 DAY 17 MEAN+S.D . 361.a 16.2 366.8 + 22.8 361.0 + 28.1 357.5 12.0 300.0 27.4 DAY IB MEAN+S.D . 377.4 15.5 381.4 23.2 379.8 + 31.0 377.5 + 20.5 312.0 + 26.7 DAY 19 MEAN+S.D . 395.6 4- 18.5 397.6 24.9 391.8 + 29.7 394.5 + 24.7 324.5 + 29.6 DAY 20 MEAN+S.D . 410.1 20.1 413.0 jf 25.2 410.2 + 34.7 413.5 34.6 334.3 + 32.5 DAY 21 MEAN+S.D . 423.0 4- 27.4 427.0 + 29.0 423.0 + 30.7 426.5 23.3 346.0 + 33.8 GRAVID UTERINE WEIGHTS (G) DAY 21C d MEAN+S.D . MEAN+S.D . 96.3 f 12.0 326.7 22.6 92.0 + 27.0 335.0 + 22.4 91.3 + 34.0 331.7 + 26.2 112.7 313.8 37.0 13.6 59.5 + 26.8 283.2 23.8 DAY = DAY OF GESTATION a. Excludes values for rats that did not have a confirmed mating date. b. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. O c. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation), d. 21C Corrected maternal body weight (day 21 of gestation body weight minus the gravid uterine weight). 418-013: PAGE B-9 C0S22 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE S (PAGE 1): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY RATS CAESAREAN -SECTIONED ON DAY 15 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV 0 (VEHICLE) 0. 1 0. 4 1. 6 RATS TESTED N 8 8 8 8 INCLUDED IN ANALYSES N 8 8 8 7a PREGNANT N6 7 8 6 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 MEAN*S.D . t29.2 + 7.7 t26.6 4 5.0 tlS.l 7.6 t5.8 4 9.0 DAYS 7 - 8 MEAN+S.D . -1.0 t .4 t0.3 3.9 tl0.2 4 7.8 t6.0 4 6.1 DAYS 8 - 9 MEANtS.D . t6.S t 3.9 t5.1 4 2.5 -1.4 _4 10.6 tl.3 4 5.5 DAYS 9 - 10 MEAN+S.D . 4.0 + 3.5 t5.7 + 11.6 t2.5 4 3.2 t5.0 + 2.4 DAYS 7 - 10 MEANtS.D . t9.5 t 5.8 tll.l + 8.9 til.4 jfr 5.0 tl2.3 8.8 DAYS 10 - 13 MEANtS.D . t15.7 t 2.1 tl0.6 4 11.0 tl7.1 + 7.6 +13.5 4.3 DAYS 13 - 15 MEANtS.D . t9.8 t 3.9 t6.8 4- 12.2 t5.9 4 9.6 +11.0 4.6 DAYS 0 - 15 MEANtS.D . t64.2 t 9.9 t55.1 _4 10.2 t4 9.5 + 12.8 +42.7 4 4.2 DAYS 0 - 15C b MEANtS.D . t44.3 t 11.7 t37.7 4 8.0 t33.2 11.6 + 25.1 4 3.2 DAYS = DAYS OF GESTATION a. Excludes values for rat 19561; necropsy and Caesarean-section observation were not recorded. b. 15C Corrected maternal body weight (day 15 of gestation body weight minus the gravid uterine weight). V 3. 2 8 8 8 +2.9 4 li. +3.1 4 8. +5.8 4 2 . +9.8 4 5. +18.6 f 8 +14.6 4 5 +3.8 4 4 +39.9 4 8 +22.8 4 7 418-013:PAGE B-10 ZS 0 0 0 Co PROTOCOL 416-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 5 (PAGE 2): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV V 0 (VEHICLE) 0.1 0.4 1.6 3. 2 RATS TESTED Na 8 8 88 INCLUDED IN ANALYSES N 7a 7b 6a ,b 4a 4 C 6a PREGNANT N7 7 5 2 6 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 MEAN+S. D . 19.4 + 9.6 +31.0 4 11.7 +14.6 4_ 7.6 +12.5 4 0.7 0.0 4 6.8 DAYS 7 - 8 MEAN+S. D . +4.8 4 10.2 +1 . 1 f 7.6 +1.6 4 4.8 +0.5 4 2.1 +1.5 4_ 3.7 DAYS B - 9 MEAN+S. D . +5.1 4 3.1 +4.3 3.4 +4.4 4 8.0 + 6.5 4 0.7 + 5.5 4 4.9 DAYS 9 - 10 MEAN+S. D . +8.0 4 2.9 4 4 . 4 4 4.9 +3.2 4 5.2 +2.5 4 4.9 +7.3 4 4 . 4 DAYS 7 - 10 MEAN+S.D . +16.0 4_ 11.0 +9.8 4 6.9 +9.2 4 7.2 +9.5 _4 7.8 +14.3 4 6.7 DAYS 10 - 13 MEAN+S.D . +12.1 4 4.0 +12.8 3.7 +15.2 4 5.5 17.5 4 2.1 11.8 _4 8.1 DAYS 13 - 15 MEAN+S.D . +6.8 4 7.4 +3.7 4 9.6 -2.6 4 15.4 +11.0 4 1.4 +5.0 4 7.3 DAYS 15 - 18 MEAN +S .D . +41.4 4 4.5 441.7 4 14.8 +50.6 4 11.6 +54.0 4 14.1 +38.8 4 11.8 DAYS 18 - 21 MEAN+S.D . 45.6 4 15.7 +45.6 4 12.2 +43.2 4 12.3 +49.0 4 2.8 + 34.0 4 8.5 DAYS 0 - 21 MEAN + S .D . +141.4 4 25.0 +144.7 4 20.3 +130.2 4_ 23.8 +153.5 4 24.7 + 104.0 4 26.3 DAYS 0 - 21C d MEAN + S .D . +45.2 18.5 + 52.7 4 16.4 + 38.9 4 14.0 +40.8 4 12.2 + 36.2 4 16.7 DAYS = DAYS OF GESTATION a. Excludes values for rats that did not have a confirmed mating date. b. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. c. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). d. 21C = Corrected maternal body weight (day 21 of gestation body weight minus the gravid uterine weight). 418-013: PAGE B-11 000324 418-013:PAGE B-12 000825 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 6 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PRECOHABITATION - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 0.1 III 0.4 RATS TESTED N 16 16 16 FEED CONSUMPTION (G) DAYS 1 - 8 DAYS 8 - 15 DAYS IS - 22 DAYS 22 - 29 DAYS 29 - 36 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 19.7 + 1.8 ( 15) a 18.8 + 1.8 I 11) a 20.0 1.7 ( 14 ]a 18.6+ 1.8 ( 12) a 18.7 + 2.0 19.4 + 1.5 18.8 + 1.6 1 15) a 19.5 + 1.6 18.9 + 1.9 ( 15) a 18.0 + 1.8 18.8 + 1.8 17.4 + 1.6 [ 15) a 18.5 + 2.1 ( 15) a 18.1 + 2.3 ( 14)a 18.0 + 1.5 DAYS 36 - 42b DAYS 1 - 42b MEAN+S.D. MEAN+S.D. 19.5 + 2.2 1 13) a 19.2 + 1.3 ( 13] a 18.0 + 1.8 18.8 + 1.4 17.9 + 1.7 [ 13) a 18.2 + 1.8 ( 13) a DAYS = DAYS OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that were incorrectly recorded, as well as those associated with spillage. b. Last value recorded before cohabitation. IV 1.6 16 18.9 + 1.5 [ 15) a 18.2 + 1.9 [ 15) a 17.9 + 1.7 I 131a 17.2+ 1.7 [ 13 )a 17.1 + 1.8 16.7 + 1.8 l 13] a 17.7 + 1.4 [ 13] a V 3.2 16 19.1 + 1.5 16.6 1.6 ( 15) a 16.4 1.5 [ 14] a 15.3 + 1.7 15.1 + 1.5 1 15)a 14.8 + 1.3 I 13] a 16.1 1.1 t 131 a PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 7 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PRECOHABITATION - SUMMARY DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 0.1 III 0.4 RATS TESTED N 16 16 16 FEED CONSUMPTION (g) DAYS 1 - 8 DAYS 8 - 15 DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 MEAN+S.D . MEAN+S.D . MEAN+S.D. MEAN+S.D . MEAN+S.D. 83.8 + 6.1 ( 151a 75.8 + 5.4 I 111a 77.0 5.1 [ 141a 69.4 + 5.2 1 12] a 68.8 + 6.2 82.1 + 6.1 75.6 + 6.0 1 15] a 75.5 + 5.8 71.4 + 6.4 [ 15] a 67.0 + 5.8 80.0 + 5.7 70.8 + 4.2 I 151 a 72.8 + 4.8 ( 15] a 68.8 + 4.8 1 141 a 67.6 + 4.7 DAYS 36 - 42b DAYS 1 - 42b MEAN+S.D. MEAN+S.D . 71.5 + 9.4 ( 131a 74.2 + 4.9 ( 131a 65.6 + 5.4 72.4 + 4.7 65.9 + 4.8 l 13] a 70.6 + 4.3 [ 131a DAYS = DAYS OF STUDY [ ] - NUMBER OF VALUES AVERAGED a. Excludes values that were incorrectly recorded, as well as those associated with spillage. b. Last value recorded before cohabitation. IV 1.6 16 80.6 + 5.9 1 15)a 74.3 + 7.6 ( IS] a 70.4 + 5.6 [ 13] a 66.1 + 5.8 1 13) a 65.6 + 6.5 64.0+ 7.9 ( 131a 70.0 + 6.0 [ 131 a V 3.2 16 81.4 + 4.9 68.7 + 5.0 l 15) a 66.6 5.2 [ 14) a 62.4 5.1 61.4 + 4.9 l 151a 61.9 + S.O [ 13] a 67.3 + 2.6 l 13) a 418-013: PAGE B-13 2S000 o PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 8 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION DOSAGE GROUP DOSAGE IMG/KG/DAY) I II III IV 0 (VEHICLE) 0. 1 0. 4 1.6 RATS TESTED N8 a 88 INCLUDED IN ANALYSES N 8 a B 7a PREGNANT N6 7 8 6 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 7 DAYS 7 - 8 MEAN+S.D . MEAN*S .D . 24.0 + 2.2 22.7 1.2 22.5 4 2.0 21.4 4 2.7 20.4 4 2.8 [ 2]b 21.0 4 2.5 b 19.5 2.8 DAYS 8 - 9 MEAN+S.D . 24.2* 1.9 24.7 4 3.4 22.1 4 2.4 22.3 + 1.6 DAYS 9 - 1 0 MEAN+S.D . 25.0 2.9 23.0 3.5 22.6 4 3.5 21.7 + 1.4 DAYS 7 - 1 0 MEANJrS .D. 24.0 + 1.7 23.0 4 3.0 21.9 2.3 21.2 * 1.7 DAYS 10 - 13 MEAN+S.D . 24.8 + 2.6 24.3 4 2.2 23.2 4 1.9 23.0 2.0 DAYS 13 - 15 DAYS 0 - 15 MEAN+S.D. MEAN+S.D. 21.9 + 2.1 23.9 + 2.0 20.7 4 1.4 ( 6]b 23.0 4 l.B I 6)b 18.9 4 3.0 l 71b 21.0 4 2.2 [ 71b 18.8 + 1.1 20.1 + 1.2 DAYS = DAYS OF GESTATION ( ) < NUMBER OF VALUES AVERAGED a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. b. Excludes values that were not recorded, as well as those associated with spillage. V 3. 2 8 8 8 16.5 4 1.7 ( 31b 16.8 4_ 4.3 20.1 4 1.4 20.5 4 2.3 19.1 4_ 1.8 22.6 4 1.7 18.7 4 2.6 l 51b 18.0 4 1.8 [ 5] b 418-013: PAGE B-14 o o N -Si PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 8 (PAGE 2): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY RATS CAESAREAN -SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV V 0 (VEHICLE) 0. 1 0. 4 I. 6 3. 2 RATS TESTED N 8 a e 8 8 INCLUDED IN ANALYSES N 7a 7b 6a, b 4a,,c 6a PREGNANT N7 7 5 2 6 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 7 DAYS 7 * 8 DAYS 8 * 9 MEAN+S.D. MEAN+S.D. MEAN+S.D . 23.6 + 1.4 21.8 + 2.6 23.6 f 2.4 23.7 2.3 23.0 3.1 25.6 + 1.8 21.0 + 3.3 l 3)d 22.2 + 2.4 22.8 4.5 20.3 + 1.0 20.5 0.7 20.0 + 4.2 17.0 + 1.8 ( 41d 15.2 3.4 I 5]d 18.2 2.7 DAYS 9 - 10 MEAN+S.D . 24.3 1.5 24 .B + 3.4 22.4 4.5 22.5 + 0.7 20.0 2.1 DAYS 7 - 10 MEAN+S.D . 23.2 1.4 24.4 f 2.3 22.5 f 3.6 21.0 + 1.4 18.0 + 2.1 DAYS 10 - 13 DAYS 13 _ 15 DAYS 15 - 18 DAYS 18 - 21 MEAN+S.D. MEAN+S.D. MEAN+S.D . MEAN+S.D . 24.4 + 1.8 22.1 f 2.5 25.3 2.1 23.9 3.2 25.3 + 2.0 20.9 + 1.6 I 6)d 24.3 + 5.6 25.1 2.6 24.5 2.8 19.1 2.9 24.9 4.7 24.9 + 2.4 23.3 1.4 21.8 1.8 26.8 f 0.2 25.5 i 0.7 20.9 f 1.9 17.6 2.0 23.6 2.4 21.9 4.2 DAYS 0 - 21 MEAN+S.D . 23.8 f 1.1 24.1 1.6 23.0 f 2.6 22.6 + 0.1 19.0 + 1.9 DAYS DAYS OF GESTATION l ) = NUMBER OF VALUES AVERAGED a. Excludes values for rats that did not have a confirmed mating date. b. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. c. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). d. Excludes values that were not recorded, as well as those associated with spillage. 418-013: PAGE B-15 0Q03Z 00 418-013: PAGE B-16 000829 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 9 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV 0 (VEHICLE) 0.1 0.4 1.6 RATS TESTED N 8 8 8 8 INCLUDED IN ANALYSES N 8 8 8 7a PREGNANT N6 7 8 6 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 DAYS 7 - 8 MEAN+S.D . MEAN*S.D . 80.6 + 8.2 73.6 + 7.3 77.2 4 6.8 70.9 4- 9.1 71.6 4 5.2 [ 2]b 72.2 4 6.8 b 7.5 4 7.6 DAYS 8 - 9 MEAN+S.D . 77.6 + 7.0 80.8 4 9.8 75.0 4 5.9 79.9 + 4.5 DAYS 9 - 10 MEAN +S .D . 78.8 + 8.1 74.0 f 11.1 76.3 4 8.6 76.7 + 4.3 DAYS 7 - 10 MEAN+S.D . 76.6 + 6.2 75.2 4 9.4 74.7 4 4.2 75.8 + 4.0 DAYS 10 - 13 MEAN+S.D . 75.8 + 6.5 76.3 4 6.3 75.7 4 3.3 78.6 + 7.5 DAYS 13 - 15 DAYS 0 - 15 MEAN+S.D. MEAN+S.D. 64.6 + 5.2 76.5 + 6.2 63.3 4 4.2 l 6) b 75.8 4 5.1 ( 6)b 60.2 4 8.6 l 7] b 71.8 4 3.8 ( 7]b 61.7 + 5.2 70.9 4.7 DAYS = DAYS OF GESTATlAl [ ] = NUMBER OF VALUES AVERAGED a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. b. Excludes values that were not recorded, as well as those associated with spillage. V 3. 2 a 8 8 67.0 4 6.6 I 31b 69.0 4 20.7 80.8 4 6.1 79.7 4 8.0 76.4 4 9.0 84.3 4 5.6 67.2 4 6.4 ( 51b 71.3 4 4.6 1 51b PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 9 (PAGE 2): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY RATS CAESAREAN -SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV V 0 (VEHICLE) 0. 1 0. 4 1. 6 3. 2 RATS TESTED N B B a 8 8 INCLUDED IN ANALYSES N 7a 7b 6a ,b 4a *C 6a PREGNANT N7 7 5 2 6 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 DAYS 7 - 8 DAYS 8 - 9 MEAN+S.D. MEAN+S.D. MEAN+S.D . 79.6 4 7.2 72.0 + 7.7 76.7 4 10.0 79.0 4 7.5 73.1 4 7.6 80.8 4 5.7 71.6 4 5.0 ( 3] d 72.2 4 7.9 73.2 4 12.7 71.6 4 3.3 71.8 4 2.6 69.1 4 14.3 70.2 4 5.7 l 4]d 61.6 4 14.6 1 5]d 73.8 4 9.7 DAYS 9 - 10 MEAN+S.D . 77.4 4_ 6.1 77.3 4 8.2 70.6 +_ 9.6 76.6 4 1.2 79.8 4 12.4 DAYS 7 - 10 MEAN+S.D . 75.3 4 5.4 77.0 4_ 5.2 71.9 4 8.7 72.4 4 4.3 73.2 4 11.1 DAYS 10 - 13 DAYS 13 _ 15 DAYS 15 - 18 MEAN+S.D . MEAN+S.D. MEAN+S.D. 75.3 5.3 66.4 4 6.7 71.4 4 5.2 76.6 4 4.6 62.3 4 4.1 ( 6]d 67.1 4_ 14.1 75.4 4^ 4.2 57.4 4 7.6 70.1 4 12.2 76.8 4 2.6 68.4 4 3.8 76.9 4 3.5 80.1 4_ 8.9 65.0 4 7.5 80.6 4 4.5 DAYS 18 - 21 MEAN+S.D. 59.4 4 6.2 62.3 4 7.0 62.1 4 6.7 63.5 4 5.8 66.4 4 11.2 DAYS 0 - 21 MEAN+S.D . 72.1 4 4.0 72.0 4_ 3 2 69.1 4 5.4 71.2 4 1.6 70.4 4 4.1 DAYS = DAYS OF GESTATION ( ) = NUMBER OF VALUES AVERAGED a. Excludes values for rats that did not have a confirmed mating date. b. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. c. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). d. Excludes values that were not recorded, as well as those associated with spillage. 418-Q13:PAGE B-17 ocsooo PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 10 (PAGE 1) : CAESAREAN -SECTIONING OBSERVATIONS - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV 0 (VEHICLE) 0.1 0.4 1.6 RATS TESTED N 8 a 8 8 INCLUDED IN ANALYSES N 8 8 8 7a PREGNANT N (%) 6( 75.0) 7( 87.5) 8(100.0) 6( 85.7) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 15 OF GESTATION N 6 1 86 CORPORA LUTEA MEAN+S.D . 17.3 + 3.4 18.0 + 1.7 17.1 + 3.0 19.8 + 3.2 IMPLANTATIONS MEAN+S.D . 15.8 + 1.9 13.4 + 3.8 13.0 + 3.7 14.2 + 1.3 VIABLE EMBRYOS N MEAN+S.D . 89 14.8 + 2.8 86 12.3 + 3.7 94 11.8 + 4.7 77 12.8 + 1.2 NONVIABLE EMBRYOS N MEAN+S.D. 6 1.0 + 1.3 8 1 . 1 + 1.3 10 1.2 + 1.4 8 1.3 + 1.0 DAMS WITH ANY NONVIABLE EMBRYOS N(%) 3( 50.0) 5( 71.4) 4( 50.0) 5 ( 83.3) DAMS WITH ALL NONVIABLE EMBRYOS N (%) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) DAMS WITH VIABLE EMBRYOS N (%) 6(100.0) 7(100.0) 8(100.0) 6(100.0) % NONVIABLE EMBRYOS/LITTER MEAN+S.D. 6.8 + 8.4 8.6 + 9.3 13.4 + 17.7 9.2 + 6.5 a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. V 3.2 8 8 8(100.0) 8 17.4 + 2.6 13.6 3.5 101 12.6 + 2.9 8 1.0 1.4 4( 50.0) 0( 0.0) 8(100.0) 6.2 + 8.3 418-013: PAGE B-18 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 10 (PAGE 2): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I II III IV V 0 (VEHICLE) 0.1 0.4 1.6 3.2 RATS TESTED N 8 8 8 8 8 INCLUDED IN ANALYSES N 7a 7b 6a, b 4a,c 6a PREGNANT DELIVERED N(%) N(%) 7(100. 0) 0( 0. 0) 7(100.0) 0( 0.0) 5( 83.3) 0{ 0.0) 2( 50.0) 0( 0.0) 6(100.0) 2( 33.3) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N 7 7 5 2 4 CORPORA LUTEA MEAN+S.D . 18.6 + 4.2 16.1 + 2.3 21.8 + 4.8 20.5 + 2.1 16.2 + 2.5 IMPLANTATIONS MEAN+S.D . 15.1 + 2.1 14.0 + 2.9 14.4 + 4.2 17.0 + 7.1 10.5 + 4.4 LITTER SIZES MEAN+S.D . 13.7 1.7 12.4 + 4.0 13.2 + 5.2 16.0 + 5.6 8.8 + 4.4 LIVE FETUSES N MEAN+S.D. 96 13.7 + 1.7 87 12.4 + 4.0 66 13.2 + 5.2 32 16.0 + 5.6 35 8.8 + 4.4 DEAD FETUSES N 0 0 0 0 0 RESORPTIONS MEAN+S.D . 1.4 + 1.7 1.6 + 1.5 1 . 2 + 1.8 1 .0 + 1.4 1.8 1.5 EARLY RESORPTIONS N MEAN+S.D . 10 1.4 + 1.7 9 1.3 + 1.1 6 1.2 + 1.8 2 1.0 + 1.4 7 1.8 + 1.5 LATE RESORPTIONS N MEAN+S.D . 0 0.0 + 0.0 2 0.3 + 0.8 0 0.0 + 0.0 0 0.0 + 0.0 0 0.0 + 0.0 a. Excludes values for rats that did not have a confirmed mating date, b. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. c. Excludes values for rat 19557, which was moribund sacrificed on day 43 of atudy (day 2 of cohabitation). 418-013: PAGE B-19 00332 PROTOCOL 41B-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 10 (PAGE 3) : CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 0.1 III 0.4 IV V 1.6 3.2 RATS TESTED N8 8 8 88 INCLUDED IN ANALYSES N 7a 7b 6a, b 4a, c 6a PREGNANT DELIVERED N (%) N(%) 7(100.0) 0( 0.0) 7(100.0) 0< 0.0) 5( 83.3) 0( 0.0) 2( 50.0) 0( 0.0) 6(100.0) 2( 33.3) RATS PREGNANT AND CAESAREAN-SECTION ED ON DAY 21 OF GESTATION N 7 7 5 24 DAMS WITH ANY RESORPTIONS N (%) 5 ( 71.4) 5( 71.4) 2( 40.0) 1( 50.0) 3( 75.0) DAMS WITH ALL CONCEPTUSES RESORBED N (%) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) DAMS WITH VIABLE FETUSES N (%) 7(100.0) 7(100.0) 5(100.0) 2(100.0) 4(100.0) PLACENTAE APPEARED NORMAL N (%) 7(100.0) 7(100.0) 5(100.0) 2(100.0) 4(100.0) a. Excludes values for rats that did not have a confirmed mating date. b. Excludes .values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. c. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). 418-013: PAGE B-20 000S33 418-013:PAGE B-21 00534 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 11 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - SUMMARY RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 0.1 III 0.4 LITTERS WITH ONE OR MORE LIVE FETUSES IMPLANTATIONS N MEAN+S.D. 7 15.1 . 2.1 7 14.0 + 2.9 5 14.4 + 4.2 LIVE FETUSES N MEAN+S.D . 96 13.7 + 1.7 87 12.4 + 4.0 66 13.2 + 5.2 LIVE MALE FETUSES N 44 46 36 % LIVE MALE FETUSES/LITTER MEAN+S.D . 46.0 + 11.7 52.2 + 14.2 59.3 + 19.4 LIVE FETAL BODY WEIGHTS (GRAMS)/LITTER MEAN+S.D . 5.12 0.29 5.31 + 0.42 5.06 + 0.09 MALE FETUSES MEAN+S.D . 5.24 + 0.29 5.50 + 0.54 5.19 + 0.12 FEMALE FETUSES MEAN+S.D. 5.02 + 0.33 5.06 + 0.33 4.84 + 0.14 PLACENTAL WEIGHTS (GRAMS)/LITTER MEAN+S.D . 0.50 + 0.06 0.54 + 0.08 0.48 + 0.06 MALE FETUSES FEMALE FETUSES MEAN+S.D . MEAN+S.D. 0.50 + 0.06 0.49 + 0.05 0.55 + 0.09 0.52 + 0.07 0.49 + 0.07 0.44 + 0.08 % RESORBED CONCEPTUSES/LITTER MEAN+S.D. 8.8 + 9.9 13.4 + 14.6 11.0 + 19.2 NO FETAL ALTERATIONS WERE IDENTIFIED AT GROSS EXTERNAL EXAMINATION ( ) = NUMBER OF VALUES AVERAGED a. Litter 19568 had no female fetuses. IV 1.6 2 17.0 + 7.1 32 16.0 + 5.6 15 45.8 + 5.9 5.02 + 0.16 5.12 + 0.20 4.95 + 0.16 0.50 + 0.06 0.52 + 0.09 0.47 + 0.04 4.6 + 6.4 V 3.2 4 10.5 + 4.4 35 8.8 + 4.4 19 55.3 31.8 4.72 + 0.23 4.82 + 0.28 4.69 + 0.27 ( 3) a 0.55 + 0.11 0.56 + 0.11 0.53 + 0.13 t 3) a 18.3 + 13.7 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 12 (PAGE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY RAT DESCRIPTION 19501 19502 19503 19504 19505 19506 1950*7 19508 19509 19510 19511 19512 19513 19514 19515 19516 DS ( 43- 66) DS ( 66 ) DG ( 17- 21) DS ( 3- 42) DG ( 0- 2) DG ( 3 ) DG ( 0- 15) DG ( 14- 15) DG ( 11- 21) NO ADVERSE FINDINGS NO ADVERSE FINDINGS LENTICULAR OPACITY DELIVERED AND SACRIFICED NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT LENTICULAR OPACITY CHROMORHINORRHEA a CHROMODACRYORRHEA a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-013: PAGE B-22 C00S35 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 12 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA 6o 0) u o DOSAGE GROUP II 0.1 MG/KG/DAY RAT DESCRIPTION 19517 19518 19519 19520 19521 19522 19523 19524 v DG ( 7- 15) DS ( 13- 42) DG ( 0- 21) DS( 10- 14) DS ( 10- 21) DS ( 22 ) DS ( 22- 30) DS ( 31 ) DS ( 30- 42) DS ( 42 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DS( 44- 45) DG ( 0 ) DG< 4- 5) DG ( 6 ) DG ( 11- 21) DS ( 43- 45) DS ( 43- 45) DG ( 0 ) DS ( 44- 45) DG ( 0 ) DG ( 0- 10) DG ( 0- 10) DG ( 0- 13) DG ( 14 ) DG ( 11- 21) TRAUMATIZED CORNEA a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a CHROMODACRYORRHEA INCISORS: MISSING/BROKEN INCISORS: GROWN IN INCISORS: MISALIGNED INCISORS: REALIGNED LOCALIZED ALOPECIA: LIMBS SNOUT: SWOLLEN CHROMODACRYORRHEA a EXOPHTHALMOS a SNOUT : SWOLLEN a LOCALIZED ALOPECIA: LIMBS a CORNEAL OPACITY RIGHT EYE: LARGE RIGHT EYE: DARK RED RIGHT EYE: LIDS UNABLE TO CLOSE MORIBUND SACRIFICED EXOPHTHALMOS EXOPHTHALMOS NO LONGER APPARENT SNOUT: SWOLLEN SWELLING RESOLVED CORNEAL OPACITY NO ADVERSE FINDINGS NO ADVERSE FINDINGS RIGHT EYE: CLOUDY, WHITE FILM SNOUT : SWOLLEN SWELLING RESOLVED EXOPHTHALMOS EXOPHTHALMOS NO LONGER APPARENT TRAUMATIZED CORNEA ' RIGHT EYE: CLOUDY, WHITE FILM LOCALIZED ALOPECIA: HEAD ALOPECIA NO LONGER APPARENT RIGHT EYE: MISSING a DS * DAY OF STUDY DG DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-013: PAGE B-23 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 12 (PAGE 3) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP II 0.1 MG/KG/DAY RAT DESCRIPTION 19525 19526 19527 19528 19529 19530 19531 19532 DG ( 2- 8) DG( 9 ) DG ( DG ( DG ( DG ( 0- 2) 3) 1- 15) 3- 15) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN INCISORS: GROWN IN NO ADVERSE FINDINGS NO ADVERSE FINDINGS EXOPHTHALMOS EXOPHTHALMOS NO LONGER APPARENT CORNEAL OPACITY TRAUMATIZED CORNEA a NO ADVERSE FINDINGS DS = DAY OF STUDY DG DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-013:PAGE B-24 00OS37 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.I2) TABLE 12 (PAGE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP III 0.4 MG/KG/DAY RAT DESCRIPTION 19533 19534 19535 19536 19537 19538 19539 19540 19541 19542 19543 19544 19545 19546 19547 19548 DG ( 0- 1) DG ( 0- 2) DG ( 0- 2) DG ( 3 ) DG ( 3- 21) DG ( 14- 21) DS ( 25 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 0 ) DG ( 10- 12) DG ( 13 ) DG ( DG ( 1) 1) LACRIMATION LENTICULAR OPACITY EXOPHTHALMOS EXOPHTHALMOS NO LONGER APPARENT CORNEAL OPACITY ENOPHTHALMOS a NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMODACRYORRHEA a EXOPHTHALMOS a SNOUT: SWOLLEN a CORNEAL OPACITY RIGHT EYE: LARGE RIGHT EYE: DARK RED RIGHT EYE: LIDS UNABLE TO CLOSE MORIBUND SACRIFICED LOCALIZED ALOPECIA: UNDERSIDE ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA UNGROOMED COAT NO ADVERSE FINDINGS NO ADVERSE FINDINGS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-013:PAGE B-25 00033 00 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 12 (PAGE 5): CLINICAL OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP IV 1.6 MG/KG/DAY RAT DESCRIPTION 19549 19550 19551 19552 19553 19554 19555 19556 19557 19558 19559 19560 19561 19562 19563 DS ( 57- 59) DS ( 60 ) DS ( 62- 63) DS ( 64 ) DS ( 44- 50) DS ( 50- 68) DS( 62- 68) DS ( 1 ) DS ( 29- 43) DG< 0- 21) DS ( 43 ) DS ( 43 ) DS ( 43 ) DS( 43 ) DS( 43 ) DS ( 43 ) DS ( 43 ) DS ( 43 ) DS( 36- 38) DS ( 39 ) DG ( 15 ) DG( 16 ) DG( 16- 21) DS( 2 ) DG ( IS- 21) DS ( 23 ) LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS NO ADVERSE FINDINGS CORNEAL OPACITY CORNEAL HEMORRHAGE a LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a CHROMODACRYORRHEA a EXOPHTHALMOS a SNOUT : SWOLLEN a CORNEAL OPACITY RIGHT EYE: LARGE RIGHT EYE: DARK RED RIGHT EYE: LIDS UNABLE TO CLOSE MORIBUND SACRIFICED EARS : SWOLLEN SWELLING RESOLVED EXOPHTHALMOS EXOPHTHALMOS NO LONGER APPARENT CORNEAL HEMORRHAGE a CHROMORHINORRHEA LOCALIZED ALOPECIA: BACK a NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS DS * DAY OF STUDY DG * DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-013:PAGE B-26 000339 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 12 (PAGE 6): CLINICAL OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP IV 1.6 MG/KG/DAY RAT ft DESCRIPTION 19564 DS( 1 ) DS ( 6 ) DS ( 29- 42) DS( 34- 35) DS ( 36- 42) DSl 31- 42) DS ( 36- 37) DS( 38 ) DG ( 0 ) DG ( 0 ) DG ( 1 ) DG ( 0- 15) DG ( 1- 15) CHROMORHINORRHEA CHROMORHINORRHEA LEFT FOREPAW: SCAB (DID NOT EXCEED 0.5 CM X 0.3 CM) RIGHT FOREPAW: SCAB (0.4 CM X 0.3 CM) RIGHT FOREPAW: TWO SCABS (DID NOT EXCEED 0.5 CM X 0.3 CM) LOCALIZED ALOPECIA: UNDERSIDE FOREPAWS: SWOLLEN SWELLING RESOLVED LEFT FOREPAW: SCAB (DID NOT EXCEED 0.5 CM X 0.3 CM) RIGHT FOREPAW: TWO SCABS (DID NOT EXCEED 0.5 CM X 0.3 CM) SCABS HEALED LOCALIZED ALOPECIA: UNDERSIDE a LOCALIZED ALOPECIA: LIMBS a DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-013-.PAGE B-27 COO540 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OP PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 12 (PAGE 7): CLINICAL OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP V 3.2 MG/KG/DAY RAT tt DESCRIPTION 19565 19566 19567 19568 19569 19570 19571 19572 19573 19574 19575 19576 19577 19578 19579 19580 DG( 0- 15) DG ( 1- 2) DG( 3 ) DS ( 13 1 DG( 0- 15) DG ( 13- 15) DS ( DS ( DS ( DG ( 9- 11) 12 ) 9- 42) 0- 21) DS( 34- 43) DS ( 36- 38) DS( 39 ) DG ( 0- 15) DG ( 21 ) DG ( 11- 21) DS ( DG ( 1) 1) DS ( 54- 68) DG ( 0- 21) DG ( 21 ) DG ( 11- IS) CORNEAL HEMORRHAGE a EXOPHTHALMOS EXOPHTHALMOS NO LONGER APPARENT CHROMODACRYORRHEA CORNEAL OPACITY LOCALIZED ALOPECIA: UNDERSIDE a NO ADVERSE FINDINGS RIGHT FOREPAW: THIRD DIGIT SWOLLEN SWELLING RESOLVED RIGHT FOREPAW: THIRD DIGIT MISSING RIGHT FOREPAW: THIRD DIGIT MISSING a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS RIGHT FOREPAW: SCAB (0.2 CM IN DIAMETER) SCAB HEALED LOCALIZED ALOPECIA: LIMBS a DELIVERED AND SACRIFICED LEFT AXILLA: MASS (1.5 CM X 1.0 CM X 1.0 CM)a NO ADVERSE FINDINGS CHROMORHINORRHEA CHROMODACRYORRHEA NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: BACK a LOCALIZED ALOPECIA: LIMBS a DELIVERED AND SACRIFICED LOCALIZED ALOPECIA: LIMBS a DS - DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 418-013:PAGE B-28 T frS O O O PROTOCOL 418-011: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 13 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a I 0 (VEHICLE) 19501 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19502 DG 15 NP 58 ALL TISSUES APPEARED NORMAL. 19503 DS 6 P 66 DELIVERED AND SACRIFICED ON DAY 66 OF STUDY. ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: 14 IMPLANTATION SITES (14 DELIVERED PUPS).b 19504 DG 21 P 65 ALL TISSUES APPEARED NORMAL. 19505 DG 21 P 64 ALL TISSUES APPEARED NORMAL. 19506 DG 21 P 64 ALL TISSUES APPEARED NORMAL. 19507 DG 21 P 67 ALL TISSUES APPEARED NORMAL. 19508 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19509 DG 21 P 64 ALL TISSUES APPEARED NORMAL. 19510 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19511 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19512 DG 21 P 66 ALL TISSUES APPEARED NORMAL. 19513 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19514 DG 15 NP 57 ALL TISSUES APPEARED NORMAL. 19515 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19516 DG 21 P 65 ALL TISSUES APPEARED NORMAL. P = PREGNANT NP = NOT PREGNANT DS = DAY OF STUDY DG DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necropsy. b. Appeared normal for their developmental ages at the time maternal death occurred. 418-013: PAGE B-29 000342 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 13 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a II 0.1 19517 DG 15 NP 58 ALL TISSUES APPEARED NORMAL. 19518 DG 21 P 64 ALL TISSUES APPEARED NORMAL. 19519 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19520 DG 0 - 43 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION PREGNANCY STATUS COULD NOT BE DETERMINED. ALL TISSUES APPEARED NORMAL. 19521 DG 21 P 66 ALL TISSUES APPEARED NORMAL. 19522 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19523 DG 21 P 67 ALL TISSUES APPEARED NORMAL. 19524 DG 21 P 66 ALL TISSUES APPEARED NORMAL. 19525 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19526 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19527 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19528 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19529 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19530 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19531 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19532 DG 15 P 57 ALL TISSUES APPEARED NORMAL. P - PREGNANT NP - NOT PREGNANT DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necropsy. 418-013:PAGE B-30 CC0S43 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 13 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a III 19533 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19534 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19535 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19536 DG 21 P 65 ALL TISSUES APPEARED NORMAL. 19537 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19538 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19539 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19540 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19541 DG 0 - 43 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION PREGNANCY STATUS COULD NOT BE DETERMINED. ALL TISSUES APPEARED NORMAL. 19542 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19543 DG 21 P 66 ALL TISSUES APPEARED NORMAL. 19544 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19545 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19546 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19547 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19548 DG 21 NP 63 ALL TISSUES APPEARED NORMAL. P = PREGNANT NP = NOT PREGNANT DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necropsy. 418-013: PAGE B-31 000544 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 13 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a 19549 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19550 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19551 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19552 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19553 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19554 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19555 DG 15 NP 57 ALL TISSUES APPEARED NORMAL. 19556 DG 21 NP 64 ALL TISSUES APPEARED NORMAL. 19557 DS 43 - 43 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION). ALL TISSUES APPEARED NORMAL. 19558 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19559 DG 21 NP 64 ALL TISSUES APPEARED NORMAL. 19560 DG 21 P 66 ALL TISSUES APPEARED NORMAL. 19561b DG 15 - 57 - 19562 DG 21 P 65 ALL TISSUES APPEARED NORMAL. 19563 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19564 DG 15 P 57 ALL TISSUES APPEARED NORMAL. P = PREGNANT HP = NOT PREGNANT DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necropsy. b. Necropsy and Caesarean-section observations were not recorded for rat 19561. 418-013: PAGE B-32 000345 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 13 (PAGE S): NECROPSY OBSERVATIONS - INDIVIDUAL DATA DOSAGE GROUP DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a V 3.2 19565 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19566 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19567 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19568 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19569 DG 21 P 66 ALL TISSUES APPEARED NORMAL. 19570 DG 15 P 58 ALL TISSUES APPEARED NORMAL. 19571 DG 21 P 66 DELIVERED ON DAY 21 OF GESTATION. ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: 15 IMPLANTATION SITES (SEVEN DELIVERED PUPS, SIX LIVE FETUSES AND TWO EARLY RESORPTIONS)-b 19572 DG 21 P 64 ALL TISSUES APPEARED NORMAL. 19573 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19574 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19575 DG 15 P 57 ALL TISSUES APPEARED NORMAL. 19576 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19577 DG 21 P 63 ALL TISSUES APPEARED NORMAL. 19578 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19579 DG 21 P 65 DELIVERED ON DAY 21 OF GESTATION. ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: 11 IMPLANTATION SITES (THREE DELIVERED PUPS AND EIGHT LIVE FETUSES).b 19580 DG 15 P 57 ALL TISSUES APPEARED NORMAL. P = PREGNANT NP = NOT PREGNANT DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necropsy. b. Appeared normal for their developmental ages at the time maternal death occurred. 418-013:PAGE B-33 OC0846 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 14 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT H DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY DAY 1 2 3 4 5 6 7 8 19501 19502 19503 19504 19505 19506 19507 19508 19509 19510 19511 19512 19513 19514 19515 19516 215. 221. 227. 231. 225 . 240. 219. 225. 221 . 236. 229. 235. 235. 234 . 223 . 221. 218. 231. 224 . 233. 224 . 236 . 222. 227. 219. 234 . 229. 232 . 238. 228. 224 . 226. 222. 234 . 224 . 235. 225. 240. 224 . 228. 218. 234 . 230. 240. 240. 238. 227. 230. 219. 240. 226. 239. 232. 243. 223 . 226. 222. 238. 236. 243. 246. 240. 222. 234 . 220. 230. 234 . 242. 238. 248. 221. 232 . 226 . 243. 242. 241 . 252. 244 . 223. 231 . 226. 240. 235. 240. 242. 253. 221. 238. 227. 238. 243. 240. 254 . 240. 229. 237. 232. 243. 233. 251. 243. 251. 226. 240. 227. 243. 241. 245. 252. 250. 233. 239. 231. 243. 238. 252. 242. 254. 231. 238. 227. 250. 251. 251. 261. 252. 230. 243 . DAY 17 18 19 20 21 22 23 24 19501 19502 19503 19504 19505 19506 19507 19508 19509 19510 19511 19512 19513 19514 19S15 19516 237. 256. 246. 272 . 253. 268. 250. 258. 246. 261 . 269. 267. 2S4 . 264 . 244 . 251 . 237. 257. 254 . 271. 257. 267. 251 . 265. 249. 261. 271. 264 . 288. 269. 244 . 258. 236. 254 . 253. 273. 261. 264 . 251. 264 . 249. 256. 266 . 270. 284 . 268. 245. 265. 239. 260. 252. 272 . 260. 268. 251 . 260. 245. 262 . 272 . 274 . 294 . 269. 249. 260 . 244 . 266. 253 . 281 . 260. 268. 259. 267. 253 . 264 . 277. 272. 300. 270. 252 . 256. 244 . 264 . 260. 280. 257. 266. 261. 272 . 257. 265. 273 . 273. 299. 275. 251. 264 . 238. 263 . 259. 282. 257. 262. 262. 268. 256 . 255. 267. 275. 292. 274 . 247. 266 . 244 . 262. 252. 277. 262. 265. 255. 267. 251. 262 . 280. 282. 297. 278 . 248 . 265. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 9 228. 240. 237. 252. 242 . 255. 234 . 241. 230. 250. 256. 248. 264 . 257. 229. 238 . 25 246 . 268. 257. 279. 264 . 268. 257. 270. 256. 264 . 280. 280 . 304 . 276. 253 . 265 . 10 223 . 244. 240. 254. 245. 255. 229. 247. 233. 252. 251. 249. 266. 247. 230. 249. 26 246. 273. 259. 285. 268. 267. 263. 273. 257. 258. 274 . 275. 303. 273 . 246 . 268. 11 227. 248. 236. 256. 246. 253. 234. 249. 236. 245. 248. 254. 264 . 249. 235. 246 . 27 245. 273. 261. 286. 272. 265. 263 . 271. 257. 259. 274 . 284 . 305. 282 . 242 . 272 . 12 232. 247. 241. 259. 247. 260. 238. 244. 231. 253 . 258. 259. 267. 256. 235. 248. 28 247. 268. 256. 284. 271. 269. 264. 268. 252. 262. 285. 284 . 310. 287. 245. 270 . 13 234. 249. 247. 264. 250. 263. 242. 252. 238. 258. 262. 255. 274 . 259. 235. 248. 29 250. 268. 260. 281. 275. 276. 260. 274 . 258. 261. 286. 282 . 311. 284 . 248. 261. 14 231. 246. 248. 263. 254. 261. 240. 256. 242 . 258. 258. 256. 276. 260. 234. 250. 30 248. 270. 263. 287. 272. 272. 268. 274. 258. 259. 281. 279. 308. 277 . 248. 268. 15 230. 251. 245. 266. 256. 263. 240. 258. 245. 251. 256. 261. 274 . 260. 235. 255. 31 244 . 268. 263 . 291. 273 . 265. 264 . 276. 260 . 257 . 277. 280. 302 . 285. 244 . 275. 16 234. 254. 241. 271. 254. 261. 246. 252. 243 . 258. 268. 263. 281. 257. 241. 254 . 32 250. 266. 259. 287. 271. 271. 267. 271. 256. 264. 288. 283 . 313 . 290 . 240 . 274 . 418-013:PAGE B-34 0084*7 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 2): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT ft DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY DAY 33 34 35 36 37 38 39 40 19S01 19502 19503 19504 19505 19506 19507 19508 19509 19510 19511 19512 19513 19514 19515 19516 251. 268. 263. 288. 281. 273. 262. 278. 258. 268. 290. 285. 317. 288. 250. 271. 252. 276. 266. 293 . 279. 266. 266. 285. 262. 265. 287. 281. 314. 283 . 248. 277. 249. 270. 267. 295. 280. 264 . 270. 283 . 260. 257. 282. 284 . 308. 286. 245. 278. 250. 266 . 263. 292. 278. 260. 262. 277. 259. 259. 291. 286. 317. 288. 246. 279. 252. 271. 265. 292. 280. 256. 262. 283 . 258. 254. 292. 286. 318. 287. 251. 274 . 252. 274 . 268. 295. 285. 257. 268. 288. 260. 260. 284 . 284 . 317, 286. 255. 278. 249. 271. 268. 296. 283. 257. 268. 283. 258. 260. 284 . 287. 312. 290. 252. 282. 251. 270. 269. 292. 282. 254. 267. 279. 260. 263. 292. 292. 322. 298. 251. 280. DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. 41 256. 273. 270. 293. 280. 261. 263. 288. 262. 261. 296. 288. 325. 293. 257. 274. 42a 255. 280. 277. 298. 285. 261. 266. 289. 266. 260. 292. 288. 324. 290. 257. 275. 418-013: PAGE B-35 000348 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 3): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT ft DOSAGE GROUP II 0.1 MG/KG/DAY DAY 19517 19518 19519 19520 19521 19522 19523 19524 19525 19526 19527 19528 19529 19530 19531 19532 12 34 5 221. 234. 225. 230. 220. 234 . 228. 233. 224 . 227. 227. 229. 226 . 228. 222. 230. 219. 238. 230. 227. 222. 236. 227. 231 . 218. 230. 233 . 231. 229. 229. 220. 230. 225. 237. 236 . 232. 229. 237. 228. 235. 225. 229. 231. 237. 227. 230. 230. 230. 228. 249. 237. 237. 235. 242. 232. 239. 229. 228. 233 . 238. 234 . 238. 233. 228. 230 250 237 240 239 247 238 239 231 234 237 240 235 239 236 234 67 227. 250. 243. 238. 235. 250 . 238. 236. 229. 238. 239. 239. 234. 242. 235. 239. 233. 252. 247. 243. 242. 245. 234. 242 . 233. 240. 244 . 245. 234 . 234. 242. 244 . DAY 17 18 19 20 21 22 23 19517 19518 19519 19520 19521 19522 19523 19524 19525 19526 19527 19528 19529 19530 19531 19532 247. 275. 264. 248. 259. 268. 260. 264 . 245. 256. 267. 255. 244 . 259. 255. 257. 245. 276. 267. 246 . 258. 266 . 259. 264 . 249. 252. 264. 254 . 243. 261. 255. 256. 250. 279. 270. 254. 264. 257. 259. 266. 247. 250. 264 . 260. 242. 254. 253 . 255. 249. 278. 265. 250. 269. 265. 262. 272 . 247. 256. 269. 260. 245. 264 . 258. 262. 253 280 266 254 267, 269. 268 270, 251. 262. 270 256 246 269, 264 268. 247. 288. 271. 253. 264 . 270. 266. 268. 255. 265. 275. 256. 248. 267. 261 . 269. 254 . 286. 274 . 258. 26B . 267. 262 . 275. 253. 260. 273 . 256 . 241. 264 . 255. 265. DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 8 237. 261. 244 . 242. 245. 256. 239. 250. 235. 241. 248. 248. 239. 240 . 244 . 245. 24 256. 284 . 274 . 258. 272. 275. 263. 277. 251. 260. 270. 266 . 246 . 264 . 262 . 265. 9 235. 264. 248. 239. 244. 255. 240. 249. 232. 239. 244 . 242. 238. 244 . 247. 239. 25 257. 289. 269. 257. 272. 282. 267. 276. 250 . 263. 272. 261. 247. 270. 265. 270. 10 232. 264. 252. 240. 242. 255. 243. 249. 234 . 246 . 252. 243 . 243. 245. 242 . 246 . 26 256. 290. 273 . 260. 266. 278. 269. 276. 247. 266. 277. 259. 246. 267. 265. 270 . 11 238. 265. 254. 240. 248. 250. 243. 254. 239. 249. 254. 248. 238. 243 . 241. 248. 27 261. 289. 278. 260. 275. 272. 269. 280. 251. 268. 278. 266. 240. 265. 262. 274 . 12 244 . 262. 252. 246. 254. 258. 245. 257. 240. 251. 260. 253. 241. 251. 243. 253. 28 260. 285. 276 . 260. 275. 278. 267. 285. 253. 264 . 277. 264 . 251 . 271. 270 . 271 . 13 244. 271. 251. 241. 251. 263. 248. 256. 240. 247. 258. 252. 244 . 255. 249. 250. 29 256. 293 . 270. 263. 271. 281. 268. 276. 252. 260. 270. 256. 246 . 271 . 271. 267. 14 240. 273. 258. 240. 250. 258. 251. 256. 242 . 243. 255. 251. 243 . 252. 248. 246. 30 249. 291. 277. 255. 268. 277. 271 . 273. 257. 268 . 275. 252 . 247. 266. 270. 270. 15 242. 272. 260. 246. 257. 257. 250. 262. 240. 250. 258. 250. 242. 250. 244 . 254. 31 258. 289. 280. 259. 276. 274 . 268. 278. 252 . 271 . 280. 261 . 244 . 263 . 262. 276. IS 249. 271. 2SB. 250. 259. 253. 254. 255. 246. 258. 262. 256. 243. 257. 253. 256. 32 261. 289. 278. 261. 280. 280. 264 . 282 . 258. 270. 278. 263. 248. 265. 270. 273 . 418-013: PAGE B-36 000849 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 4): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT DOSAGE GROUP II 0.1 MG/KG/DAY DAY 33 34 35 36 37 38 39 40 19517 19518 19519 19520 19521 19522 19523 19524 19525 19526 19527 19528 19529 19530 19531 19532 262. 294. 273. 265. 279. 285. 271. 282 . 253. 262 . 278. 257. 251. 274 . 273. 272. 256. 298. 277. 265. 274. 286. 273. 276. 254. 262. 276. 253. 247. 270. 269. 277. 259. 295. 280. 269. 281. 276. 274 . 280. 254. 268 . 282. 260. 242. 267. 266. 281. 262. 296. 276. 272. 284. 280. 266. 279. 251. 270. 286. 261. 247. 271. 277. 278. 260. 301. 273. 267. 280. 286. 266. 278. 257. 269. 286 258 249. 274. 283 . 271. 255. 303. 269. 264. 279. 285. 268. 276. 252. 268. 284 . 256. 250. 272. 276. 271. 262. 300. 276. 265. 288. 278. 273 . 281. 254 . 271. 279. 262. 244 . 267. 273. 282 . 266. 298. 278. 268. 288. 282. 274. 284. 255. 270. 283. 266. 254. 270. 280. 278. DAY DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. 41 42a 265. 303. 278. 273. 285. 287. 275. 280. 252. 272 . 285. 259. 253. 277. 282. 280. 257. 310. 276. 263. 288. 282 . 274. 279. 255. 272. 293. 257. 251. 269. 276 . 269. 418-013: PAGE B-37 000350 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 5) : BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT 8 DOSAGE GROUP Ill 0.4 MG/KG/DAY DAY 19533 19534 19535 19536 19537 19538 19539 19540 19541 19542 19543 19544 19545 19546 19547 19548 DAY 12 221 . 234 . 221 . 225. 217. 230. 226 . 230. 218. 237. 234. 231. 222. 234 . 223 . 228. 224. 241 . 220. 225. 219. 227. 227. 229. 218. 236. 233 . 231. 220. 235. 228. 231. 17 18 3 222. 244 . 226. 230 . 223 . 229. 225. 230. 216. 231. 242. 232. 232 . 231 . 228 . 233 . 19 4 220. 251. 223. 233. 221. 233. 230. 234. 220. 240. 244 . 235. 231. 239. 230. 234 . 20 5 226 256 226 235 222 237 231 236 225 245 249 241 231 240 235 239 21 6 7 8 9 10 11 12 13 14 236. 258. 230. 230. 224 . 237. 230. 236. 226. 244 . 245. 245. 229. 239. 232. 242 . 228. 253. 227. 230. 227. 241. 234. 233. 227. 240. 252. 246 . 234 . 242. 234 . 246. 229. 262. 229. 242. 233 . 242. 238. 241. 229. 250. 254. 248. 238. 248 . 243. 246 . 223 . 266. 230. 242. 227. 243. 233 . 243 . 230 . 252 . 254 . 245. 239. 248. 242. 243 . 227. 263. 230. 243. 229. 243. 230. 243. 230. 253. 253 . 254. 240. 248. 240. 247. 233 . 259. 233. 240. 235. 244 . 233. 241. 228. 248. 257. 253. 235. 250. 239. 249. 231. 264. 230. 243. 233 . 248. 236 . 246. 234 . 250. 257. 258. 239. 258. 247. 250. 232. 266. 230 . 246. 231. 246. 239. 250. 236. 257. 263. 257. 241. 263. 248. 253 . 230. 269. 232. 246. 231. 247. 238. 250. 236. 261. 258. 253. 245. 265. 246. 254. 22 23 24 25 26 27 28 29 30 15 16 235. 267. 235. 245. 234 . 248. 236. 249. 236. 258. 266. 259. 248. 263. 247. 258. 236. 276. 234 . 246. 237. 250. 240 . 251. 231. 255. 268. 262. 246. 267. 251. 259. 31 32 19533 19534 19S35 19536 19537 19538 19539 19540 19541 19542 19543 19544 19545 19546 19547 19548 240. 282. 239. 246. 238. 252. 242. 256. 235. 261. 266. 265. 244 . 269. 249. 261. 237. 280 . 238. 250. 234 . 253. 242. 256. 240. 265. 265. 265. 247. 274 . 252. 260. 238. 281. 242 . 253 . 235. 252 . 244 . 253 . 239. 264 . 276. 262. 253. 278 . 249. 261. 244 . 286. 243 . 255. 237. 255. 246. 264 . 238 . 259. 281. 269. 253. 274 . 251. 266 . 246 291 244 252 244 254 248 263 239 266 274 273 254 283 258 268 245. 288. 248. 255. 243 . 252 . 248 . 263 . 244 . 269. 277. 276. 255. 288. 259. 267. 246. 281. 244 . 259. 236. 253 . 246 . 260. 241 . 268. 281 . 274 . 257. 286 . 254 . 264 . 243. 293 . 247. 255. 239. 255. 248 . 266 . 242 . 266. 285. 271. 259. 285. 257. 266 . 245. 296. 245. 255 . 238. 255. 253. 274 . 236. 266 . 287. 284 . 260 . 288. 263 . 269. 248. 297. 247. 258. 238. 256. 250. 270. 242. 272. 284 . 278. 261. 292 . 262 . 268. 252. 295. 251. 261 . 244 . 256 . 247. 269. 244 . 272 . 289. 280. 260 . 295 . 259. 268. 250. 303. 251. 260. 243. 256. 250. 275. 246 . 264. 290 . 275. 263. 293 . 263 . 266. 244 . 298. 254. 256. 240. 254 . 253 . 280. 239. 272. 284 . 270. 262 . 286 . 267. 265. 247. 296. 255. 262. 240. 254. 249. 274 . 243. 274. 278 . 278 . 265. 294 . 265 . 268. 253. 291. 258. 265. 240. 256. 251. 273. 248 . 272. 282 . 281 . 260 . 297. 267. 264 . 257. 302. 256 . 261. 245. 259. 253. 277. 247. 265. 290. 276. 260. 293 . 269. 263. M DAY DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013:PAGE B-38 00085 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 6): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT tt DOSAGE GROUP Ill 0.4 MG/KG/DAY DAY 33 34 35 36 37 38 39 19533 19534 19535 19536 19537 19538 19539 19540 19541 19542 19543 19544 19545 19546 19547 19548 254. 302. 254. 262. 246. 258. 256. 280. 242 . 271. 28S . 274. 268. 291. 269. 264. 252. 300. 254. 264. 247. 258. 254. 281. 243. 278. 282. 276 . 267. 299. 268. 268. 256. 299. 258. 264. 247. 259. 252. 280. 244 . 274 . 289. 283 . 264. 300. 273. 266 . 259. 308. 257. 262. 253. 259. 258. 282. 246. 267. 291. 289. 260. 298. 272. 264 . 260. 308. 262. 259. 252 261. 258 289. 243. 274 294 282 264 295 274 261 257. 305. 263. 266. 250. 260. 259. 281. 248. 276. 288 . 284 . 272. 299. 271. 260. 256. 302. 261. 266. 248. 258 . 257. 278. 244 . 274. 293. 275. 272. 308. 271. 261. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Laat value recorded before cohabitation. 40 261. 308. 262. 264 . 255. 256 . 257. 284 . 249. 269. 296. 290. 270. 305. 275. 264. 41 264. 307. 259. 262. 253. 260. 257. 289. 247. 273 . 291. 295. 270. 300. 275. 267. 42a 259. 311. 259. 268. 254. 260. 255. 286. 246 . 276. 286. 211. 290. 274 . 297. 266. 418-013:PAGE B-39 00035 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 7): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT tt DOSAGE GROUP IV 1.6 MG/KG/DAY DAY 1 2 3 4 19549 19550 19551 195S2 19553 19554 19555 19556 19557 19558 19559 19560 19561 19562 19563 19564 220. 239. 228. 230. 218. 232. 231. 230. 221. 234 . 220. 225. 220. 235. 216. 231. 220. 236. 230. 230. 216. 236. 234. 229. 225. 239. 227. 229. 220. 239. 220. 230. 219. 247. 230. 235. 215. 234. 237. 232. 224 . 239. 228. 232. 222. 242. 221. 229. 226. 250. 234. 242. 221. 239. 234. 240. 223 . 244 . 230. 230. 228. 243. 225. 230. DAY 17 18 19 20 19549 19550 19551 19552 19553 19554 19555 19556 19557 19558 19559 19560 19561 19562 19563 19564 248. 271. 265. 245. 232. 266. 266. 258. 251. 266 . 242. 257. 242 . 258. 243 . 255. 246. 267. 270. 240. 238. 267. 263. 251. 247. 266. 244 . 253. 239. 256 . 247. 253 . 248. 265. 272. 244 . 237. 263. 268. 260. 246. 268. 245. 255. 236. 259. 246 . 254 . 250. 271. 273. 247. 236 . 267. 272. 263 . 251. 271. 242. 260. 241. 256. 243. 257. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 5 229 251 240 236 228 247 240 238 226 247 227 232 232 239 229 240 21 252 273 273 246 235 271 272 266 253. 269 246. 260 244 . 264 . 242 . 260 . 6 227. 251. 242. 232. 226. 248. 249. 233 . 230. 250. 235. 238. 224 . 236. 230. 240. 22 252. 275. 275. 243. 240. 271. 270. 264 . 254 . 276. 250. 256. 245. 264. 245. 259. 7 228. 253. 242. 237. 223. 246. 248. 240. 234 . 246. 237. 243. 218. 241. 229. 242 . 23 248. 268. 279. 244 . 236. 266. 275. 264 . 252. 273 . 248. 259. 241 . 267. 248. 258. 8 231. 264. 252. 240. 224. 250. 248. 246. 236. 249. 234. 246. 230. 241. 234. 242 . 24 253 . 274. 284 . 249. 238. 269. 276. 268. 254 . 273. 244 . 264 . 247. 266. 246. 256. 9 233. 260. 250. 239. 233. 254. 246 . 243. 231. 252. 235. 240. 237. 240. 233 . 241. 25 254. 275. 280. 249. 233 . 271. 277. 261 . 256 . 276 . 248. 262 . 249. 262 . 247 . 261. 10 233. 263 . 249. 231. 230. 254 . 250. 240. 238. 256. 239. 246. 235. 240. 235. 247. 26 253. 272. 278. 246. 234. 269. 273. 262. 255. 277. 247. 256 . 244 . 266. 246. 261. 11 230. 256. 250. 239. 232. 251. 253. 247. 239. 256. 237. 247. 233. 245. 229. 245. 27 251. 266. 279. 248. 239. 267. 282. 272. 257. 280. 248. 260. 242 . 266. 246 . 259. 12 235. 264 . 251. 243. 230. 258. 258. 253. 243. 258. 233. 252. 234. 249. 233. 250. 28 254. 272. 281. 252. 238 . 271. 284 . 267. 256 . 282 . 242 . 262. 245. 267. 249. 259. 13 237. 270. 261. 242. 233 . 260. 253 . 251. 243. 259. 238. 248. 242. 247. 239. 251. 29 249. 275. 274. 247. 232. 271. 282 . 261 . 249. 280. 246. 257. 247. 262 . 249. 254 . 14 238. 269. 263 . 238. 236. 263. 2S6 . 246. 241. 263. 242. 248. 239. 250. 240. 252. 30 251. 272. 279241. 234 . 266. 275. 257. 252 . 284 . 248 . 250 . 242 . 262 . 253 . 258 . IS 238. 263. 264 . 242. 236. 259. 260. 254. 244 . 267. 242. 255. 231. 250. 239. 252. 31 250. 265. 283 . 243. 241. 265. 283 . 260. 251. 283. 246. 256. 240. 268. 252. 257. 16 241. 269. 263. 245. 232. 264. 264 . 257. 247. 263. 238. 255. 239. 253. 240. 252. 32 256. 271. 284 . 249. 239. 271. 285. 263. 251. 280. 243. 266. 243 . 269. 253. 254. Co 418-013: PAGE B-40 00035 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 8) : BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT H DOSAGE GROUP IV 1.6 MG/KG/DAY DAY 33 34 35 36 37 38 39 40 19549 19550 19551 195S2 19553 19554 19555 19556 19557 19558 19559 19560 19561 19562 19563 19564 256. 272. 280. 250. 237. 273. 284 . 257. 252. 282. 248. 261. 246 . 266. 254. 249. 255. 274. 284. 252. 228. 272. 280. 254 . 252. 281. 250. 259. 246. 263. 251. 256 . 252. 266. 284. 254. 231. 267. 287. 261. 255. 284. 249. 265. 242. 266. 253. 256. 258. 272. 284 . 257. 235. 272. 287. 261. 255. 288. 247. 266. 246. 268. 258. 2S4 . 259. 272. 280. 257. 231. 272. 290. 258. 259. 288. 252. 265. 249 269. 253 253. 263. 270. 278. 260. 226. 268. 282. 250. 256. 290. 254. 260. 252. 271. 257. 250 . 266. 266 . 283. 262. 227. 268. 288. 258. 252. 291. 255. 266. 244 . 268. 260. 257. 264. 269. 283. 262. 232. 271. 288. 258. 254. 294. 252. 264. 249. 268. 258. 256. DAY DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. 41 42a 269. 271. 283. 261. 233. 273. 2B6 . 256. 255. 291. 254. 259. 246. 273 . 260. 256. 272. 270. 282. 263 . 230. 270. 280 . 253. 251. 290. 252. 258. 239. 263. 255. 250. 418-013: PAGE B-41 00035 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 9): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT ft DOSAGE GROUP V 3.2 MG/KG/DAY DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 19565 19566 19567 19568 19569 19570 19571 19572 19573 19574 19575 19576 19577 19578 19579 19580 217. 238. 234 . 240. 220 . 228. 235. 227. 221 . 236. 230. 230. 222. 232. 220. 230. 222. 234. 237. 246. 221 . 234 . 230. 227. 224 . 237. 233 . 233 . 224 . 232. 221. 230. 214. 240 . 239. 247. 224 . 234 . 228. 226. 221. 237. 230. 234. 225. 235. 223. 228. 221. 246 . 241. 249. 226. 234 . 243. 226. 221. 244 . 237. 236. 226. 242 . 224. 238. 225. 245 245 . 248. 229. 233 . 241 228 226 249 238 242. 226 245 221 , 242 . 226 . 240. 240. 24 7. 236 . 237. 245. 230. 231. 249. 245. 237. 229. 244 . 224. 240 . 224 . 243 . 231. 250 . 233 . 238. 249. 231. 233. 245. 242 . 243 . 229. 250. 227. 237. 222. 249. 236. 247. 238. 243. 252. 234 . 231. 254. 248 . 249. 231. 254. 225. 237. 224 . 251. 237. 248. 236. 242 . 252. 233. 230. 255. 247. 242 . 235. 252. 222. 243 . 224 . 242 . 232. 2S0. 234 . 238. 248 . 235. 236. 255. 248 . 238. 236. 252. 224 . 242. 232. 240. 230. 251. 236 . 243. 253. 238. 234. 253. 245. 247. 238. 256. 217. 234. 229. 245. 234 . 256. 238. 246. 255 . 241. 233. 254. 245. 248. 240. 258. 215. 241. 224 . 252 . 235. 255. 236 . 247. 252. 240. 232. 258. 249. 247. 239. 258. 215. 238. 229. 247. 230. 253. 238. 247. 250. 242 . 230. 260. 252. 247. 242. 257. 223. 237. 228. 243 . 229. 250. 238. 247. 256. 242. 238. 257. 252. 250. 240. 258. 223. 236. 231. 250. 230. 253. 239. 247. 256. 242 . 235. 256. 245. 254 . 241. 261. 215. 238. DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 19565 230. 227. 236 . 230. 232 230. 227. 229. 232 . 231. 227. 231. 236. 235. 232. 231. 19566 255. 253 . 246. 252. 257, 253. 241. 249. 251. 251. 243. 252 . 249. 249. 239. 245. 19567 234 . 230. 227. 233 . 235 229. 226 . 232. 230. 228. 223 . 230. 231. 226 . 220. 225. 19568 258. 262. 258 . 264 . 265. 266. 262. 261. 263. 267. 264. 263 . 264 . 264. 265. 264. 19569 237. 241. 241. 241. 243 245. 241. 237. 246. 247. 242. 238. 240 . 242. 239. 243 . 19570 247. 248 . 251. 250. 254 255. 249. 252. 249. 250. 254 . 251. 251. 249. 249. 254 . 19571 256 . 252. 257 . 255. 258 254 . 257. 256. 258. 252 . 256. 255. 250. 240. 244 . 244 . 19572 243 . 243 . 242 . 242 . 242 243. 241 . 242. 241. 244. 245. 248. 245. 245. 242. 241. 19573 235. 235. 234 . 241. 242 242. 238. 235 . 238. 240. 241. 237. 237. 238 . 239. 237. 19574 258. 254 . 258. 258. 256 258 . 258 . 259. 252 . 247. 253. 254 . 250 . 242. 249. 249. 19575 247. 254 . 255. 256 . 256 254 . 255. 260. 261 . 257. 255. 256 . 256. 253 . 248 . 244 . 19576 254 . 250. 253 . 250. 252 253. 259. 260. 259. 253. 261. 257. 257. 253 . 266. 260. o 19577 19578 241. 256. 243 . 242. 239. 241 257 . 261 . 262 . 261 241. 259. 238. 262. 234 . 232. 232. 262. 260 . 259. 228. 227. 222. 224 . 226. 225. 264 . 264 . 258 . 249. 255 . 257. o 19579 212. 219. 215. 213. 221 220. 220. 220. 218 . 213 . 215. 214 . 213. 214 . 211. 210. o 19580 239. 238. 238. 245. 244 243 . 239. 247. 243 . 245 . 241. 247. 244 . 242 . 237. 242 . 03 07 07 DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013: PAGE B-42 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 10): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA RAT ft DOSAGE GROUP V 3.2 MG/KG/DAY DAY 33 34 35 36 37 3B 39 19565 19566 19567 19568 19569 19570 19571 19572 19573 19574 19575 19576 19577 19578 19579 19580 233 . 246. 224 . 264 . 246. 252. 244 . 238. 238. 248 . 249. 257. 227. 256. 211. 244 . 234 . 248. 224. 263. 245. 250. 243 . 235. 241. 245. 250. 257. 225. 252. 209. 242. 233. 236. 221. 267. 240. 247. 252. 236. 243. 249. 245. 265. 227. 254. 208 . 239. 233 . 242 . 222. 266 . 244 . 250. 244 . 239. 243 . 251. 241. 268. 226. 257. 210. 240. 232. 249. 223. 267 , 241 249. 244 237 238 251 246 261 227 258 214 237 233 . 243. 226. 267. 241. 250. 239. 234 . 240. 248. 243. 261. 226. 251. 217. 235. 227. 236. 224. 265. 238. 247. 243. 236. 237. 247. 245. 262. 224. 253. 214. 234. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. 40 41 42a 231. 236. 222. 269. 239. 245. 242. 243. 240. 250. 246. 264. 224 . 254. 216. 238. 232. 244. 225. 268. 238. 246. 241. 240. 236 . 251. 243 . 261. 224 . 251. 212. 237 . 225. 244. 223. 268. 234. 249. 241. 245. 235. 240. 233. 255. 221. 249. 211. 231. S9000 418-013: PAGE B-43 O PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295..12) TABLE IS (PAGE 1): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA RAT DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 ii 12 19501 P 19502 NP 19503 P 19504 P 19505 P 19506 P 19507 P 19508 P 19509 P 19510 P 19511 P 19512 P 19513 P 19514 NP 19515 P 19516 P 257. 263 . 269. 270. 274 . 278. 280. 276. 288 . 294. 297 . 300 . 304. 303. NO CONFIRMED MATING DATE ; DELIVERED ON DAY 66 OF STUDY 295. 306 . 308. 308. 311. 313. 317. 281 . 294 . 301. 301. 302. 308. 309. 266 . 273 . 279. 284 . 287. 295. 297. 272 . 2B4 . 286. 290. 291. 298. 296 . 291. 297. 306. 313 . 314. 313 . 315. 269. 273 . 273. 284 . 287. 293. 299. 259. 267. 278. 279. 290. 291. 301. 291 . 299. 305. 307. 311. 311. 316. 306 . 314 . 312. 316. 323. 318. 324. 330. 329. 334 . 340. 350. 349. 355. 290. 298. 302. 301. 311. 313 . 316. 258. 267. 274 . 274 . 278. 283. 285 . 282. 288. 298. 306 . 302 . 309. 311. 277. 307. 316. 306. 294 . 289. 319. 299. 302. 322. 318. 355. 321. 286. 285. 285. 306 . 318. 309. 301. 287. 318. 293. 302. 310. 322. 353 . 321. 287. 311. 289. 308. 320 . 316 . 304 . 296 . 325. 302. 311. 321. 324. 353 . 325. 295. 315. 295. 316. 323. 323 . 313. 305. 333 . 306 . 311. 324 . 326. 360. 326. 295. 323. 302. 316. 329. 332. 312. 310. 339. 311. 325. 329. 330. 367. 330. 302. 328. 306. 316. 343. 328. 318. 310. 344 . 317. 322. 334. 333 . 369. 337. 308. 334. DAY 13 14 15 16 17 18 19 20 21 GRAVID UTERINE WEIGHTS 19501 P 19502 NP 19503 P 19504 P 19505 P 19506 P 19507 P 19508 P 19509 P 19510 P 19511 P 19512 P 19513 P 19514 NP 19515 P 19516 P 311 . 310. 315. 318. 314 . 310. NO CONFIRMED MATING DATE; DELIVERED ON DAY 66 OF STUDY 342. 347. 348 . 357. 368. 385. 396. 333 . 337. 343. 356. 374 . 390. 415. 325. 326. 335. 340. 360. 376. 397 . 312. 316. 304 . 323 . 335. 350. 363. 350. 349. 364 . 322. 324 . 328. 333. 345. 363 . 380 . 330. 338. 339. 338. 345. 351. 336. 341 . 344 . 354 . 376. 388. 406 . 374 . 380. 386. 335. 337. 342. 309. 313. 316 . 334. 347. 350. 357. 375. 390. 412. 418. 433 . 412. 371. 399. 416 . 422. 430. 452. 431. 394. 377. 431 . 446 . 16.56 79.93 118.23 99.38 93.66 21.37 86.56 15.76 27.22 96.29 21.00 17.04 99.80 P PREGNANT NP * NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) Si DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013: PAGE B-44 00085 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PAGE 2): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA RAT * DOSAGE GROUP II 0.1 MG/KG/DAY PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 11 12 19517 NP 19518 P 19519 P 19520 19521 P 19522 P 19523 P 19524 P 19525 P 19526 P 19527 P 19528 P 19529 P 19530 P 19531 P 19532 P 277 . 308. 276. 247. 292. 292. 288 . 296. 259. 278. 281. 258. 258. 271. 270. 281. 276. 268. 270. 272 . 272. 271. 272. 272. 273 . 272. 274 . 316. 319. 320. 322. 329. 333. 338. 334. 334. 339. 346. 280. 288. 287. 296. 296. 302. 304 . 307. 316. 320. 328. MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 299. 308. 309. 303. 306. 315. 306. 313 . 321. 320. 321. 295. 305. 305. 308. 318. 320. 320. 319. 328. 328. 332. 294 . 304 . 310. 309. 312. 315. 330. 317. 318. 332. 334 . 304 . 311. 319. 318. 327. 327. 323. 331 . 336. 342. 346. 261. 271 . 272 . 274 . 277. 278. 281. 281. 285. 283 . 287. 280. 288. 287. 291. 291. 294. 301. 303 . 306. 308. 313 . 293 . 294 . 296. 300. 302. 310. 310. 303. 310. 340. 318. 275. 285. 290. 300. 300. 300. 308. 315. 317. 317. 330. 265. 273. 268. 280. 282. 282. 284 . 284 . 289. 292. 302. 285. 293. 287. 296. 297. 312. 307. 309. 313. 317. 323. 270. 281. 280 . 290. 292. 294 . 296. 296 . 298. 308. 312 . 293 . 298. 297. 296. 299. 306. 303 . 309. 316. 312. 317. 281. 347. 332. 328. 338. 336. 348. 291. 317. 326. 327. 298. 329. 323. 328. DAY 13 14 15 16 17 18 19 20 21 GRAVID UTERINE WEIGHTS 19517 NP 279. 273 . 281. 19518 P 354 . 357. 359. 370 . 385 . 392. 412. 424 . 448. 71.83 19519 P 333. 338. 339. 361 . 379. 397. 414. 429 . 433. 115.99 19520 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19521 P 336. 335. 339. 353. 370. 389. 402. 413. 429. 101.43 19522 P 339. 346 . 355. 18.40 19523 P 339. 346. 325. 353 . 368. 378. 390. 405. 411. 88.34 19524 P 351. 361. 367. 373 . 393 . 411. 430. 450. 470. 117.23 19525 P 292. 297. 303 . 8.80 19526 P 318. 322. 324 . 15.42 19527 P 328. 335. 342. 25.35 o o 19528 P 19529 P 19530 P 330. 309. 333 . 337. 308. 335. 329. 320 . 314 . 318. 327. 329. 344 . 343 . 360. 355. 380. 370. 400. 378. 420 . o 19531 P 328. 332. 333. ta 19532 P 332. 332. 347. 42.75 106.57 16.30 21.41 16.54 Cl CO P PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013:PAGE B-45 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PAGE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA RAT ft DOSAGE GROUP III 0.4 MG/KG/DAY PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 11 12 19533 P 19534 P 19535 P 19536 P 19537 P 19538 P 19539 P 19540 P 19541 19542 P 19543 P 19544 P 19545 NP 19546 P 19547 P 19548 NP 267. 270. 273 . 273. 275. 274. 278. 276. 281. 278. 282. 289. 323. 325. 333. 329. 333. 336. 342. 336. 342 . 345. 341. 349. 253. 244. 260. 266. 267. 270. 274 . 268. 278. 281. 281. 285. 274 . 275. 282. 285. 286. 289. 289. 290. 293. 295. 296. 304. 257. 257. 266 . 265. 268. 269. 272. 257. 264. 266. 267. 275. 260. 267. 268. 274 . 272. 278. 279. 269. 279. 280. 285. 289. 262. 261 . 271. 277. 280. 284 . 286. 279. 292. 298. 296. 303. 285. 234 . 291 . 301. 302. 310. 310. 315. 303. 308. 319. 320. 320. MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 284 . 304 . 292. 316. 294. 318. 294 . 319. 297. 323. 302. 328. 306 . 327. 304. 331. 303. 331. 304. 333. 310. 338. 317. 328. 290. 304 . 314. 310. 308. 306. 313. 315. 327. 313. 320. 328. MATING NOT CONFIRMED 296. 286. 300. 307. 310. 312. 317. 304. 298. 316. 326. 331. 272. 285. 288. 292 . 292. 296. 290. 289. 315. 294 . 296. 302. 270. 276. 279. 280. 284 . 283 . 283 . 267. 284. 288. 288. 287. 288. 356. 293 . 310. 279. 297. 306. 330. 324. 349. 330. 333. 314. 292. DAY 13 14 15 16 17 18 19 20 21 GRAVID UTERINE WEIGHTS 19533 P 19534 P 19535 P 19536 P 19537 P 19538 P 19539 P 19540 P 19541 19542 P 19543 P 19544 P 19545 NP 19546 P 19547 P 19548 NP 294. 298. 298. 306. 323. 338. 363. 367. 334 . 378. 380 . 400 . 303. 305. 312. 315. 320. 317. 338. 353. 371. 281. 284 . 285. 301. 302. 291. 310. 321. 319. 331. 340. 335. MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; 343. 334 . 339. 353. 361. 364 . 375. 396. 419. 329. 333. 345. MATING NOT CONFIRMED 334 . 339. 333. 350. 353. 371. 314 . 323. 333 . 296. 291. 286. 284 . 284 . 284. 352. 405. 397. 371. 438. 418. 387. 445. 429. PREGNANCY STATUS COULD NOT BE DETERMINED 430. 447. 458. 375. 285. 377. 291. 396. 294 . 85.28 94.87 9.30 123.08 6.02 18.47 19.11 20.45 .... 19.46 116.27 16.14 36.94 21.57 P - PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013: PAGE B-46 00035 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PAGE 4): MATERNAL BODY HEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA RAT If DOSAGE GROUP IV 1.6 MG/KG/DAY PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 19549 NP 19550 P 19551 NP 19552 NP 19553 P 19554 P 19555 NP 19556 NP 19557 19558 P 19559 NP 19560 P 19561a 19562 P 19563 P 19564 P MATING NOT CONFIRMED 276. 276. 279. 279. 278. 284 . 290. 266. MATING NOT CONFIRMED MATING NOT CONFIRMED 245. 245. 252. 254 . 256 . 253 . 255. 256. 282. 275. 282. 278. 280. 282. 286. 282. 292. 293 . 300. 299. 301. 300. 295. 296. 267. 268. 261. 264 . 266. 264 . 258. 262. MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 286. 287. 302. 305. 303 . 308. 299. 297. 260. 260. 264 . 267. 265. 269. 268. 268. 274 . 278. 283. 286 . 284 . 291. 288. 286. 254. 260. 262. 259. 264 . 265. 262. 268. 272. 277. 282. 283 . 283 . 288. 292. 285. 2S6 263 . 273 . 267. 270. 266. 267. 266. 258. 257. 264. 266. 268. 268. 273 . 271. DAY 13 14 15 16 17 18 19 20 19549 NP 19550 P 19551 NP 19552 NP 19553 P 19554 P 19555 NP 19556 NP 19557 19558 P 19559 NP 19560 P 19561a 19562 P 19563 P 19564 P MATING NOT CONFIRMED 311. 314 . 322. MATING NOT CONFIRMED MATING NOT CONFIRMED 286. 287. 292. 307. 318. 323. 298. 296. 299. 268. 263. 261. 268. 270. 265 272 . MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 324 . 337. 332. 267. 259. 250. 252. 254 . 252 258. 309. 308. 319. 328. 349. 363 377 . 260. 262. 263. 316. 326. 328 . 350. 366. 392 412 . 278. 285. 295. 287. 292. 295. 276. 258. 389. 438. P - PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Necropsy and Caesarean-section observations were not recorded for rat 19561. 8 9 10 11 12 281. 291. 294. 303. 312. 259. 286. 287. 263. 309. 271. 285. 262. 287. 265. 274 . 21 261. 288. 293 . 261. 302. 273 . 291. 266 . 294 . 264. 276. 269. 296. 287. 259. 275. 299. 294 . 267. 277. 303. 296 . 266. 306. 276. 290. 268. 300. 268. 279. 317. 279. 295. 271. 306 . 276 . 283. 324. 273. 296. 265. 313. 279. 288. GRAVID UTERINE WEIGHTS 17.57 270. 257. 410. 443 . 18.42 17.90 19.53 86.60 138.89 16.06 16.16 03S000 418-013: PAGE B^7 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE IS (PAGE 5): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA RAT ft DOSAGE GROUP V 3.2 MG/KG/DAY PREGNANCY STATUS DAY 0 1 2 3 4 5 19565 P 19566 P 19567 P 19568 P 19569 P 19570 P 19571 P 19572 P 19573 P 19574 P 19575 P 19576 NP 19577 P 19578 NP 19579 P 19580 P 234 . 224 . 227. 244 . 252. 249. 228. 226. 228. 259. 263. 264 . 249. 247 . 248. 257. 257. 258. 245. 247. 249. 250. 255. 252 . 229. 225. 223 . 252 . 253. 256. 243 . 239. 239. MATING NOT CONFIRMED 230. 229. 230. MATING NOT CONFIRMED 219. 216. 213. 237. 235. 242. 228. 251, 230. 266 . 247. 260. 253 . 255. 227, 261. 228, 227 211. 235 229. 246. 231. 266. 244 . 253. 256. 253 . 238. 261. 239. 219. 213 . 240. 230. 244. 234. 270. 252. 266. 255. 257. 244 . 261. 242. 208. 227. 244 . DAY 13 14 15 16 17 18 19565 P 19566 P 19567 P 19568 P 19S69 P 19570 P 19571 P 19572 P 19573 P 19574 P 19575 P 19576 NP 19577 P 19578 NP 19579 P 19580 P 259. 264 . 261. 278. 286. 285. 269. 278. 266. 288. 291. 290. 264 . 266. 278. 294. 302. 291. 288. 293 . 297. 282. 290. 276. 268. 273 . 275. 285. 288. 291. 294 . 292. 299. MATING NOT CONFIRMED 247. 245. 248. MATING NOT CONFIRMED 240. 248 . 250 . 266. 270. 275. 296 287 304 300 258 258 312. 306. 332. 317. 315. 319. 346. 330. 264 . 269. 274 . 288. P PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 6 234. 244. 237. 270. 252. 265. 259. 259. 247. 266. 250. 208. 223 . 244 . 19 328. 334. 357. 349. 280. 299. 7 219. 245. 236. 264. 248. 260. 245. 259. 250. 245. 256. 219. 217. 236 . 20 341. 346. 369. 360. 286 . 304. 8 234. 241. 245. 270. 245. 265. 245. 262. 243. 254. 250. 217 . 222. 240. 21 351. 349. 385. 376. 295. 320. 9 240. 249. 250. 275. 249. 272. 259. 261. 248. 262. 252. 221. 229. 245. 10 11 12 250. 266. 254. 276. 262. 275. 268. 264. 256. 275. 269. 251. 270. 256. 284 . 256. 281. 275. 277. 260 . 275. 268. 257. 271. 261. 287. 265. 282. 278. 275. 266. 279. 277. 230. 236. 240. 238 . 251. 236. 266. 236. 260. GRAVID UTERINE WEIGHTS 7.98 15.63 17.59 41.84 86.30 20.43 78.32 17.44 17.60 19.43 31.64 20.54 9S 000 418-013: PAGE B^8 PROTOCOL 418-013! ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 1): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA RAT DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY DAYS 1- 8 8- IS 15- 22 22- 29 29- 36 36- 41 41-- 42a 19501 19502 19503 19504 19505 19506 19507 19508 19509 19510 19511 19512 19513 19514 19515 19516 131. 131. 138. 144 . 144 . 147. 141. 124 . 123 . 161. 149. 126. b 156. 120. 130. 114 . 123. 145. 147. b b b 131. 128. b b 124 . 149. 144. 120. 122. 129. 128. 150. 152. 140. 147. b 144 . 127. b 150. 131. 167. 140 . 138 . 123. b 119. 128. 140 . b 133. 146 . 129. 121. b 150. 133. b 141. 107. 120. 124. 107. 141. 141. 143. 122. 148. 130. 120. 132. 140. 120. 158. 131. 106 . 135. 92. 81. 100. 95. 90. b 97. b b b 99. 93. 114. 92. 88. 87. 18. 19. 23. 20. 22. 24. 19. 21. 23. 18. 22. 16 . b b c 14 . DAYS DAYS OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. c. Value was incorrectly recorded and was excluded from group averages and statistical analyses. 418-013: PAGE B-49 000S6 w PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 16 (PAGE 2) : FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA RAT ft DOSAGE GROUP II 0. 1 MG/KG/DAY DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36 - 41 41- 42a 19517 19S18 19519 19520 19521 19522 19523 19524 19525 19526 19527 19528 19529 19530 19531 19532 127. 143. 128. 122. 148. 138. 120. 140. 121. 140. 145. 137. 146. 129. 154 . 133. 116. 139. 128. 115. 143. 126. b 150. 119. 148. 138. 124. 131. 135. 142. 121. 119. 137. 130. 125. 149. 125. 150. 149. 123. 146. 144 . 128. 127. 145. 153 . 133. 126. 135. 128. 132. 152. b 119. 153. 108. 148. 124. 126. 117. 140. 144 . 136 . 109. 129. 125. 132. 143. 124 . 120. 127. 114 . 151. 121. 111. 112. 128. 143 . 135. 87. 98. 83 . 87. 115. 87. 83. 99. 73 . 97. 82. 84 . 88. 94. 102 . 85. 11. 21. 16. 15. 16. 19. 18. 18. 17 . 18. 23 . 16. 20. 19. 20. 14 . DAYS DAYS OF STUDY ALL HEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. 418-013: PAGE B-50 SSGOO C4 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 3) : FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA RAT 8 DOSAGE GROUP Ill 0. 4 MG/KG/DAY DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36 - 41 41- 42a 19533 19534 19535 19536 19537 19538 19539 19540 19541 19542 19543 19544 19545 19546 19547 19548 125. 151. 115. 124 . 130. 129. 117. 122 . 112 . 144 . 149. 138. 130. 146. 127. 146 . 108. 131. 103. 123. 118. 115. 105. 122. 112. 135. 138. 135. 123. c 124 . 131. 122. 152. 122. 125. 118. 118. 114. 132. 117. 132. c 140. 128. 168. 126. 134. 113. 141. 117. 122. 109. 117. 110. 133. 112 . c 150. 133. c 162. 134 . 123. 126. 129. 118. 121. 122. 115. 115. 133. 127. 123. 126. 130. 124. 158. 139. 118. 88. 94. 84 . 79. 80. B7. 79. 87. 85. 87. 101. 97. 94 . c 96. 78. 17. 23. 15. 20. 20. 17. 15. 18. b c c 24. 19. 21 . 21. 16 . DAYS = DAYS OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Value was incorrectly recorded and was excluded from group averages and statistical analyses. c. Spilled fqed precluded the calculation of this value. 418-013:PAGE B-51 0QOS6 H*4 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 4): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA RAT DOSAGE GROUP IV i .6 MG/KG/DAY DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36 - 41 41- 42a 19549 19550 19551 19552 19553 19554 19555 19556 19557 19558 19559 19560 19561 19562 19563 19564 127. 146. 141. 121. 131. 141 . b 127. 124 . 144 . 137. 132. 109. 140. 143. 120. 121. b 142. 103. 145. 147. 123. 130. 124. 143. 122. 122. 118. 119. 138. Ill. b 123. 137. 108. 116. b 141. 135. 124 . 140. b 115. 119. 129. 137. 109. 118. 107. 126. 105. b b 138. 125. 111. 140. b 118. 116. 123. 134 . 105 . 124. 108. 125. 104. 122. 139. 122. Ill. 10B. 140. 115. 121. 111. 120. 142. 105. 92. 75. 74. 82. b 96. 89. 77. 72. 98. 92. 85. 87. 84 . 105. 77. 18. IS. 16. 17. 17. 18. 13. 15. c b b 12. 10. 16. 21. 9. DAYS = DAYS OF STUDY ALL HEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. c. Value was incorrectly recorded and was excluded from group averages and statistical analyses. SSGOd 418-013: PAGE B-52 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 5) : FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA RAT # DOSAGE GROUP V 3 .2 MG/KG/DAY DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36 - 41 41- 42a 19565 19566 19567 19568 19569 19570 19571 19572 19573 19574 19575 19576 19577 19578 19579 19580 118. 143. 125. 132. 141. 139. 140. 117 . 132. 157. 144 . 141. 126. 135. 128 . 127. 109. 119. 101. 123. 117. 128. 122. 118. 113. b 125. 121. 128. 123. 90. 103. 102. 124. 105. 125. 114. 144 . 115. 111. 110. b 114. 116. 103. Ill. b 111. 104 . 106. 101. 117. 100. 120. 112. 112. 98. 117. 115. 122. 76. 116. 91. 112 . 100. 103. 97. 114. 113. 118. 95. 93. 117. 113. 97. 126. 90. 103. b 106. 68. 78. 72. 81. 67. 84. 73. 73. 73 . b 77. 82. 68. 77. 80. 74 . 11. 17. 14 . 18. 10. 17. 15. 19. b b c 9. 10. 15. 13 . 13 . DAYS DAYS OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. c. Value was incorrectly recorded and was excluded from group averages and statistical analyses. 418-013: PAGE B-53 00QS66 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA RAT H DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 19501 P 19502 NP 19503 P 19504 P 19505 P 19506 P 19507 P 19508 P 19509 P 19510 P 19511 P 19512 P 19513 P 19514 NP 19515 P 19516 P 17. 20. 22. 22. 24 . 23. 22. 21. 22. 22. 23. 22. 23. 23. NO CONFIRMED MATING DATE ; DELIVERED ON DAY 66; o f STUDY 23. 24. 22. 22. 27. 21. 24 . 22. 21. 24 . 23 . 24 . 25. 22. 24 . 29. 28. 23 . 25. 26. 23 . 27. 32. 27. 21. 28. 25. 20. 21. 23. 26 . 24 . 23. 24 . 24. 20. 22. 23. 23. 24 . 27. 22. 16. 25. 23. 26. 45. 30. 27. 18. 23. 23. 24 . 25. 23. 26. 24 . 21. 23 . 24 . 21. 24 . 19. 22. 21. 25. 29. 27 28. 29. 21 . 13 . 21. 24 . 24 26. 27. 22. 21. 22. 22. 23 23 . 24. 20. 26. 22 . 24 . 24 24 . 17. 24 . 21. 25. 22. 23 . 17. 21. 21. 23. 22. 21. 24. 24 . 22. 24. DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 19501 P 19502 UP 19503 P 19504 P 19505 P 19506 P 19507 P 19508 P 19509 P 19510 P 19511 P 19512 P 19513 P 19514 NP 19515 P 19516 P 23. 18. 19. 16. NO CONFIRMED MATING DATE ; DELIVERED ON DAY 66 OF STUDY 25. 20. 25. 28 . 27 24 . 33. 26. 23. 23 . 29. 28 28. 27 . 21. 25. 22. 25. 28 25. 22. 21. 17. 22. 27. 28 27. 16 . 23 . 25. 20. 25. 18. 20. 25 23. 24. 25. 18. 21. 17. 20. 17. 28. 24 . 24 23. 25. 26. 23. 26. 22 . 24 . 20. 27. 23. 24 . 31. 26 30. 26. 26. 24. 23. 23. 11. 17. 25. P PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 8- 9 22. 19. 24 . 23. 27. 25. 25. 25. 27. 22. 21. 25. 28. 24. 20. 9 - 10 1 0 - 1 1 11 - 12 12 - 13 23 . 23. 18. 24 . 24 . 22. 21. 24. 24 . 28. 28. 23. 26. 25. 26. 25. 26 . IB. 28. 24 . 24. 24. 24. 22. 25. 25. 29. 26. 25. 25. 26. 23. 29. 28. 26. 29. 22. 21. 22. 24. 22. 19. 23 . 22. 28. 26. 26. 29. 28. 26 . 27. 29. 23. 23. 27. 21. 23. 25. 28. 26. 418-013: PAGE B-54 000367 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 2) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA RAT K DOSAGE GROUP II 0.1 MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 1 0 1 0 - 1 1 11 - 12 12 - 13 19517 NP 1951B P 19519 P 19520 19521 P 19522 P 19523 P 19524 P 19525 P 19526 P 19527 P 19528 P 19529 P 19530 P 19531 P 19532 P 21. 12. 16. 19. 18. 15. 18. 16. 16. 14. 17. 26. 24 . 26. 23. 24 . 28. 25. 25. 25. 23. 26. 19. 23. 15. 23. 21. 24 . 27. 20. 25. 24. 26. MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 25. 23. 25. 19. 21. 22. 15. 24 . 24. 25. 22. 17. 23 . 20. 22. 23 . 23 . 24. 19. 25. 22. 23. 22. 21. 25. 23. 24. 24. -21. 22. 25. 25. 24. 23 . 24 . 29. 27. 29. 28. 25 . 26 . 27. 32. 29. 16. 21 . 19. 21. 19. 19. 22 . 20. 19. 18. 20. 20. 25. 23. 26. 23. 23. 28. 22 . 24. 25. 23. 20 . 19. 20. 21 . 18. 24. 26 . 18. 22. 19. 19. 25. 24 . 28. 28. 26. 26. 30. 26. 29. 24. 30. 21. 23. 20. 26. 22. 21. 24. 18. 24. 21. 22. 22. 27. 23. 29. 25. 30 . 24 . 26. 2B . 27. 25. 16 . 21 . 24 . 28. 24 . 24 . 25. 22. 28. 26. 26. 20. 23 . 20. 23. 23 . 25. 22. 23 . 27. 24. 22. 21. 27. 21. 23. 24. 26. 26. 21. 24. 27. 24 . 20. 24 . 28. 25. 18. 25. 26. 25. 2S . 26. 27. 23. 26. 20. 30. 26. 28. 29. 28. DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 19517 NP 19518 P 19519 P 19520 19521 P 19522 P 19523 P 19524 P 19525 P 19526 P 19527 P 19528 P 19529 P 19530 P 19531 P 19532 P 11 . a 22. a 27. 25. 32. 28. 28. 28. 21. 22 . 26. 28. 25. 29. 22 . 12. MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION;; PREGNANCY STATUS COULD NOT BE DETERMINED 20. 17 . 28. 29. 25. 26 . 26 . 18. 21 . 22. 26 . 18 . 21. 23. 26. 28 . 24 . 20. 24 . 22. 27. 26. 28 . 30. 28. 26 . 22. 18 . 25. 18 . 19. a 21 . 19. 4 . 16. 16. 28. 26. 23 . 20. 21. 27. 26. 25. 31. 24 . 23 . 23. 15. 22. 17. 24 . 21. P = PREGNANT NP * NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL HEIGHTS WERE RECORDED IN GRAMS (G) . a. Spilled feed precluded the calculation of this value. e s s G i) 418-013: PAGE B-55 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA RAT DOSAGE GROUP III 0.4 MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 1 0 1 0 - 1 1 11 - 12 12 - 13 19533 P 19534 P 19535 P 19536 P 19537 P 19538 P 19539 P 19540 P 19541 19542 P 19543 P 19544 P 19545 NP 19546 P 19547 P 19548 NP 19. 17. 17. 20. 19. 19. 19. 19. 18. 18. 21. 18. 22. 24 . 27. 23. 24. 22. 26. a 21. 23 . 25. 24. 25. 29. 8. 17. 17. 21. 18. 20. a 20. 21. 22. 20. 21. 27. 16. 21. 21. 21. 21. 17. 21. 22. 19. 17. 25. 21. 22. 16 . 16 . 19. 21. 22. 20. a 18. 21. 20. 19. 21. 23. 19. 18. 19. 18. 17. 22. 16. 18. 20. 19. 18. 24. 19. 18. 17. 20. 22. 20. 22. a 25. 22. 20. 22. 22. 23 . 17. 24 . 20. 24 . 23 . 25. a 21. 27. 24. 22. 22. 28. MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 25. 20. 22. 21. 24 . 24. 20. 21. 20. 23. 24. 26. 26. 27. 23. 25. 26. 23. 26 . 24. 24. 25. 26. 25. 26. 28. 24. 28. 19. 25. 24. 22. a 24. 24. 30. 21. 27. 28. MATING NOT CONFIRMED 12 . 22. 24 . 27. 26. 30. a 25. 29. 26. 25. 24. 32. 21. 23. 23 . 25. 21. 23. a 21. 22. 23. 23. 23. 27. 16. 21. 20. 21. 20. 17. a 21. 23 . 22. 23. 20. 24 . DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 19533 P 19534 P 19535 P 19536 P 19537 P 19538 P 19539 P 19540 P 19541 19542 P 19543 P 19544 P 19545 NP 19546 p 19547 P 19548 NP 19. 16. 22. 25. 25. 23. 22. 19. 26. 11. 24 . 17. 17. 25. 30. 23. 22. 19. 21. 10. 21. 20. 25. 26 . 28. 24 . 20 . 17. 17. 9. 22. 18. 22. 14 . MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION;; PREGNANCY STATUS COULD NOT BE DETERMINED 22. b 23. 19. 39. 27. 28. 23. 28. 20. 24 . 21. MATING NOT CONFIRMED 26 . 20. 31 . 21. 31. 34 . 27. 21 . 22. 18. 20. 19. 18 . 18. 20 . 18. 20 . 19. P PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS - DAYS OF PRESUMED GESTATION ALL HEIGHTS WERE RECORDED IN GRAMS (G) . a. Value not recorded. b. Spilled feed precluded the calculation of this value. 418-013:PAGE B-56 ssooo PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 4): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA RAT 8 DOSAGE GROUP IV 1.6 MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4-5 5-6 6-7 7- 8 19549 NP 19550 P 19551 NP 19552 NP 19553 P 19554 P 19555 NP 19556 NP 19557 19558 P 19559 NP 19560 P 19561c 19562 P 19563 P 19564 P MATING NOT CONFIRMED 14. 16. 16. 16. 18. 20. a MATING NOT CONFIRMED MATING NOT CONFIRMED 15. 16. 20. 21. 18. 17. a b 17. 15. 19. 23. 22. a 19. 20. 18. 24 . 18. 16. a 19. 11 . 18. 16. 15. 11. 17. MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 14 . 22. 22. 22. 20. 18. a 15. 18. 18. 19. 20. 21. 21. 24 . 23 . 21. 22. 23. 18. 16. 18. 20. 18. 17. 20. 18. a 18. 21 . 19. 20. 21. 21. 17. 19. 23 . 21. 21. 19. 18. a 13 . 18. 16. 20. 18. 20. a 17. 19. 23. 17. 18. 23. 20. 21. 19. 20. 18. 17. DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 19549 NP 19550 P 19551 NP 19552 NP 19553 P 19554 P 19555 NP 19556 NP 19557 19558 P 19559 NP 19560 P 19561c 19562 P 19563 P 19564 P MATING NOT CONFIRMED 20. 16. MATING NOT CONFIRMED MATING NOT CONFIRMED 20. 18. 21. 18. 17 . 17 . 14 . b 21. 20. 14. 19. 18. MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 23. 15. 8. b 13. 14 . 14. 17. 20. 17. 24 . 28. 26. 27. 27. 29. 18 . 13 . 25. 21 . 22 . 33. 25. 28. 26. 22. 18. 19. 15. 15. 17. 22. 21. P PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . a. Value not recorded. b. Spilled feed precluded the calculation of this value. c. Necropsy and Caesarean-section observations were not recorded for rat 19561. 8- 9 23. 22. 22. 19. 18. 25. 20. 17. 20. 23. 22. 20. 9 - 10 10 - 11 11 - 12 12 - 13 22. 23. 25. 21. 21. 23. 24. 28. 21. 22. 19. 26. 15. 20. 20. 23. 15. 19. 22. 16. 24. 25. 23. 27. 21. 23. 20. 16. 22. 21. 20. 26. 20. 21. 15. 13. 23. 25. 25. 23. 22. 22. 23. 24. 20. 20. 17 . 22. 418-013: PAGE B-57 000370 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE S) : MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA RAT DOSAGE GROUP V 3.2 MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 19565 P 19566 P 19567 P 19568 P 19569 P 19570 P 19571 P 19572 P 19573 P 19574 P 19575 P 19576 NP 19577 P 19578 NP 19579 P 19580 P 6. 9. 12. 10. 15. 14. a 26. 19. 17. 17. 13. 10. 13. 14. 14 . 10. 15. 17. 15. 18. 21. 20. 19. 13. 16. 17. 17. 17. 17. a 19. 16. 14 . 15. 13. 19. 17. 15. 10. 16. 15. 15. 14 . 22. 22. 23. 14. 18. 15. 19. 22. 19. 18. 11. 16. 19. 18. 18 . 13. 18. 17. 33. 14. 4 . 8. b 19. 22. 20. a 15. 15. 18. 21. 23. 18. 21. a 18. 7. 12. b b 19. 20. a 13. MATING NOT CONFIRMED 13 . 14. 11. 6. 2. 6. a b MATING NOT CONFIRMED 9. 13 . 6. 30. 27. 16. 10. 17. 9. 16. 15. 13 . 18. 15. a 15. DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 19565 P 19566 P 19567 P 19568 P 19569 P 19570 P 19571 P 19572 P 19573 P 19574 P 19575 P 19576 NP 19577 P 19578 NP 19579 P 19580 P 18. 14 . 20. b 22. b 21. 17. 20. 14. 17. 24. 21. b 21. 19. 25. 18 . 14 . 25. 22. 16. 20. 19. 25. 20. MATING NOT CONFIRMED 18. 20. 23 . MATING NOT CONFIRMED 17. 15. 19. 18. 15. 23. 22. 27. 25. 17. 23. 25. 26. 24 . 23. 24. 17. 21. 11. 25. 29. 24 . 23. 31. 26. 23. 30. 22. 24 . 23. 24 . 18 . 15. 18. 19. P - PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS DAYS OF PRESUMED GESTATION ALL HEIGHTS HERE RECORDED IN GRAMS (G) . a. Value not recorded. b. Spilled feed precluded the calculation of this value. 8- 9 21. 20. 22. 20. 14. 21. 21. 20. 20. 20. 20. 18. 16. 17. 9-10 19. 22. 22. 19. 18. 19. 22. 18. 19. 23. 23. 20. 23. 17. 10 - 11 11 - 12 12 - 13 21. 20. 23. 21. 27. 27. 21. 24. 24. 21. 19. 24. 21. 19. 20. 17. 22. 24. 21. 24. 22. 19. 24 . 20. 21. 20. 24. 24. 22. 27. 22. 22. 28. 23. 21. 26. 19. 16. 18. 21. 20. 21. 418-013: PAGE B-58 00371 PROTOCOL 418 013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 18 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA RATS CAESAREAN-SECTION ON DAY 15 OF PRESUMED GESTATION VIABLE EMBRYOS NONVIABLE EMBRYOS RAT K RIGHT LEFT HORN TOTAL RIGHT LEFT HORN TOTAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 19501 19502 19508 19510 19511 19513 19514 19515 5 9 14 NOT PREGNANT 9 7 16 5 6 11 8 11 19 10 6 16 NOT PREGNANT 7 6 13 101 000 112 000 000 213 DOSAGE GROUP II 0.1 MG/KG/DAY 19517 19522 19525 19526 19527 19530 19531 19532 NOT PREGNANT 8 7 15 426 5 5 10 8 9 17 4 7 11 8 7 IS a 4 12 000 011 000 011 404 101 101 DOSAGE GROUP III 0.4 MG/KG/DAY 19535 19537 19538 19539 19540 19542 19544 19547 437 033 8 4 12 7 6 13 9 7 16 8 8 16 6 5 11 5 11 16 112 123 000 202 000 000 213 000 IMPLANTATION SITES RIGHT LEFT HORN TOTAL 6 9 15 9 7 16 6 7 13 8 il 19 10 6 16 9 7 16 8 7 15 437 5 5 10 8 10 18 8 7 IS 9 7 16 9 4 13 549 156 8 4 12 9 6 15 9 7 16 8 8 16 8 6 14 5 11 16 CORPORA LUTEA RIGHT LEFT OVARY TOTAL 9 10 19 9 7 16 6 7 13 11 12 23 10 6 16 10 7 17 9 8 17 7 9 16 10 9 19 10 10 20 9 9 18 9 11 20 10 6 16 8 7 15 4 8 12 9 6 15 9 9 18 11 7 18 9 10 19 12 10 22 6 12 18 418-013:PAGE B-59 coosv: PO PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 18 (PAGE 2): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA RATS CAESAREAN-SECTION ON DAY 15 OF PRESUMED GESTATION VIABLE EMBRYOS NONVIABLE EMBRYOS IMPLANTATION SITES RAT 8 RIGHT LEFT HORN TOTAL RIGHT LEFT HORN TOTAL RIGHT LEFT HORN TOTAL DOSAGE GROUP IV 1.6 MG/KG/DAY 19550 19553 19554 19555 19558 19561 19563 19564 9 5 14 000 9 5 14 6 7 13 10 1 7 7 14 9 5 14 011 9 6 15 NOT PREGNANT 1 6 13 303 10 6 16 NECROPSY AND CAESAREAN-SECTION OBSERVATIONS WERE NOT RECORDED 5 7 12 112 6 8 14 7 4 11 011 7 5 12 DOSAGE GROUP V 3.2 MG/KG/DAY 19565 19566 19567 19570 19573 19574 19575 19580 246 7 5 12 8 5 13 6 9 15 S 9 14 7 6 13 7 6 13 7 8 15 000 10 1 000 112 011 000 044 00 0 246 8 5 13 8 5 13 7 10 17 5 10 15 7 6 13 7 10 17 7 8 15 CORPORA LUTEA RIGHT LEFT OVARY TOTAL 9 10 19 10 10 20 10 7 17 10 8 18 8 11 19 13 13 26 6 7 13 9 8 17 9 8 17 7 10 17 5 10 15 10 10 20 8 11 19 11 10 21 418-013: PAGE B-60 a o o (a si PROTOCOL 418-013: ORAL ((7WVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 18 (PAGE 3): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON1 DAY 21 OF PRESUMED GESTATION VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 19503 NO CONFIRMED MATING DATE ; DELIVERED ON DAY 66 OF STUDY 19504 8 4 5 7 12 0 0 0 2 3 5 0 0 0 7 10 17 10 13 23 19505 8 9 4 13 17 0 0 0 0 2 2 0 0 0 4 15 19 4 16 20 19506 4 9 s 8 13 0 0 0 1 0 1 0 0 0 6 8 14 7 a 15 19507 7 7 9 5 14 0 0 0 0 1 1 0 0 0 9 6 15 13 12 25 19509 5 7 6 6 12 0 0 0 1 0 1 0 0 0 7 6 13 7 8 15 19512 7 7 4 10 14 0 0 0 0 0 0 0 0 0 4 10 14 4 11 15 19516 5 9 6 8 14 0 0 0 0 0 0 0 0 0 6 8 14 8 9 17 DOSAGE GROUP II 0.1 MG/KG/DAY 19518 6 5 6 5 11 0 0 0 2 0 2 0 0 0 8 5 13 8 7 15 19519 9 7 9 7 16 0 0 0 0 0 0 0 0 0 9 7 16 10 7 17 19520 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19521 7 8 9 6 15 0 0 0 0 0 0 0 0 0 9 6 15 9 6 15 19523 3 7 8 2 10 0 0 0 1 1 2 1 1 2 10 4 14 10 4 14 19524 12 4 8 8 16 0 0 0 0 1 1 0 0 0 8 9 17 9 12 21 19528 3 2 1 4 5 0 0 0 I 2 3 0 0 0 2 6 8 6 9 15 19529 6 8 7 7 14 0 0 0 1 0 1 0 0 0 8 7 15 8 8 16 DOSAGE GROUP h i 0.4 MG/KG/DAY 19533 8 4 4 8 12 0 0 0 0 0 0 0 0 0 4 a 12 4 12 16 19534 8 6 7 7 14 0 0 0 0 0 0 0 0 0 7 7 14 13 15 28 19536 5 13 6 12 18 0 0 0 0 0 0 0 0 0 6 12 18 7 12 19 19541 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19543 11 6 10 7 17 0 0 0 1 1 2 0 0 0 11 a 19 11 10 21 19545 MATING NOT CONFIRMED 19546 4 1 4 1 5 0 0 0 1 3 4 0 0 0 . 5 4 9 14 11 25 19548 NOT PREGNANT M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-013: PAGE B-61 CC0374 PROTOCOL 418-013! ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 18 (PAGE 4)i CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT O M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL DOSAGE GROUP IV 1.6 MG/KG/DAY 19549 MATING NOT CONFIRMED 19551 MATING NOT CONFIRMED 19552 MATING NOT CONFIRMED 19556 NOT PREGNANT 19557 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19559 NOT PREGNANT 19560 5 7 9 3 12 0 0 0 0 0 0 0 0 0 9 3 12 12 7 19 19562 10 10 8 12 20 0 0 0 0 2 2 0 0 0 8 14 22 8 14 22 DOSAGE GROUP V 3.2 MG/KG/DAY 19568 6 0 6 0 6 0 0 0 0 3 3 0 0 0 6 3 9 8 11 19 9569 6 7 9 4 13 0 0 0 0 0 0 0 0 0 9 4 13 10 6 16 19571 DELIVERED ON DAY 21 OF GESTATION 19572 6 6 7 5 12 0 0 0 1 2 3 0 0 0 a 7 15 8 9 17 19576 MATING NOT CONFIRMED 19577 1 3 4 0 4 0 0 0 1 0 1 0 0 0 5 0 5 8 5 13 19578 MATING NOT CONFIRMED 19579 DELIVERED ON DAY 21 OF GESTATION M > MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. 418-013: PAGE B-62 000875 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 19 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 15 OP GESTATION NUMBER OF LIVE EMBRYOS RAT (I TOTAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 19501 19502 19508 19510 19511 19513 19514 19515 14 NOT PREGNANT 16 11 19 16 NOT PREGNANT 13 DOSAGE GROUP II 0.1 MG/KG/DAY 19517 19522 19525 19526 19527 19530 19531 19532 NOT PREGNANT 15 6 10 17 11 15 12 DOSAGE GROUP III 0.4 MG/KG/DAY 19535 19537 19538 19539 19540 19542 19544 19547 7 3 12 13 16 16 11 16 CONCEPTUSES ..... NONVIABLE N N% 15 i 6.7 16 0 0.0 13 2 15.4 19 0 0.0 16 0 0.0 16 3 18.8 15 0 0.0 7 1 14.3 10 0 0.0 18 1 5.6 15 4 26.7 16 1 6.2 13 1 7.7 9 2 22.2 6 3 50.0 12 0 0.0 15 2 13.3 16 0 0.0 16 0 0.0 14 3 21.4 16 0 0.0 418-013: PAGE B-63 000876 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 19 (PAGE 2): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA RATS CAESAREAN- SECTIONED ON DAY 15 OF GESTATION NUMBER OF LIVE EMBRYOS -- CONCEPTUSES ----NONVIABLE RAT # TOTAL N N% DOSAGE GROUP IV 1.6 MG/KG/DAY 19SS0 195S3 19554 19555 19558 19561 19563 19564 14 14 0 0.0 13 14 1 7.1 14 IS 1 6.7 NOT PREGNANT 13 16 3 18.8 NECROPSY AND CAESAREAN -SECTION OBSERVATIONS HERE NOT RECORDED 12 14 2 14.3 11 12 1 8.3 DOSAGE GROUP V 3.2 MG/KG/DAY 19565 19566 19567 19570 19573 19574 19S75 19580 6 12 13 IS 14 13 13 15 6 0 0.0 13 1 7.7 13 0 0.0 17 2 11.8 15 1 6.7 13 0 0.0 17 4 23.5 15 0 0.0 418-013: PAGE B-64 CQ0377 PROTOCOL 418-013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.I2) TABLE 19 (PAGE 3): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION NUMBER OF LIVE FETUSES AVERAGE FETAL BODY WEIGHT (G) --- CONCEPTUSES ----RESORBED RAT (t MALE FEMALE TOTAL MALE FEMALE TOTAL1 a N N% DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 19503 19504 19505 19506 19507 19509 19512 19516 NO CONFIRMED DATE OF MATING; DELIVERED ON DAY 66 OF STUDY 8 4 12 4.75 4.44 4.64 8 9 17 5.13 5.10 5.12 4 9 13 5.66 5.48 5.54 7 7 14 5.47 5.21 5.34 5 7 12 5.34 5.14 5.22 7 7 14 5.14 4.87 5.01 5 9 14 5.17 4.87 4.98 17 19 14 15 13 14 14 5 29.4 2 10.5 1 7.1 1 6.7 1 7.7 0 0.0 0 0.0 DOSAGE GROUP II 0. 1 MG/KG/DAY 19518 19519 19520 19521 19523 19524 19528 19529 6 5 11 4.80 4.52 4.67 13 2 15.4 9 7 16 5.29 5.16 5.23 16 0 0.0 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 7 8 15 5.12 4.79 4.94 15 0 0.0 3 7 10 5.47 5.28 5.34 14 4 28.6 12 4 16 5.51 5.11 5.41 17 1 5.9 3 2 5 6.45 5.06 5.89 a 3 37.5 6 8 14 5.87 5.54 5.68 15 1 6.7 DOSAGE GROUP III 0 .4 MG/KG/DAY 19533 8 4 12 5.30 4.89 5.16 12 0 0.0 19534 8 6 14 5.00 4.82 4.92 14 0 0.0 19536 5 13 18 5.31 5.05 5.12 18 0 0.0 19541 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19543 11 6 17 5.18 4.72 5.02 19 2 10.5 19545 MATING NOT CONFIRMED 19546 4 1 5 5.18 4.70 5.09 9 4 44.4 19S48 NOT PREGNANT O .......... ............................ ........................................ ^ a. TOTAL = SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES. o C3 oo 418-013:PAGE B-65 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 19 (PAGE 4): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION NUMBER OF LIVE FETUSES AVERAGE FETAL BODY WEIGHT (G) --- CONCEPTUSES -.... RESORBED RAT tt MALE FEMALE TOTAL MALE FEMALE TOTAL a N N% DOSAGE GROUP IV 1.6 MG/KG/DAY 19549 19551 19552 19556 19557 19559 19560 19562 MATING NOT CONFIRMED MATING NOT CONFIRMED MATING NOT CONFIRMED NOT PREGNANT MORIBUND SACRIFICED ON DAY 43 OP STUDY (DAY 2 OF COHABITATION) NOT PREGNANT 5 7 12 5.26 5.06 5 1 4 12 10 10 20 4.97 4.84 4.91 22 0 0.0 2 9.1 DOSAGE GROUP V 3.2 MG/KG/DAY 19568 19569 19571 19572 19576 19577 19578 19579 6 0 6 4.50 6 7 13 4.84 DELIVERED ON DAY 21 OF GESTATION 6 6 12 4.78 MATING NOT CONFIRMED 1 3 4 5.18 MATING NOT CONFIRMED DELIVERED ON DAY 21 OF GESTATION -- 4.50 4.57 5.00 4.50 4.66 4.68 5.04 9 13 15 5 3 33.3 0 0.0 3 20.0 1 20.0 a. TOTAL - SUM OF FETAL HEIGHTS/NUMBER OF LIVE FETUSES. 418-013:PAGE B-66 o o o U3 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 1): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY IS OF PRESUMED GESTATION EMBRYO K 1 2 3 4 S 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY RAT tt CLs 19S01 9/10 E A A A A A / A A A A A A A A A 19S02 NOT PREGNANT 19508 9/ 7 A A A A A A A A A / A A A A A A A 19510 6 / 7 E A A A A A / A A A A A E A 19511 11/12 A A A A A A A A / A A A A A A A A A A A 19513 10/ 6 A A A A A A A A A A / A A A A A A 19514 NOT PREGNANT 19515 10/ 7 E A A E A A A A A / A A E A A A A DOSAGE GROUP II 0 .1 MG/KG/DAY 19517 NOT PREGNANT 19522 9/ 8 A A A A A A A A / A A A A A A A 19525 7/ 9 A A A A / E A A 19526 10/ 9 A A A A A / A A A A A 19527 10/10 A A A A A A A A / A A E A A A A A A A 19530 9/ 9 E A E A A E A E / A A A A A A A 19531 9/11 A A A A A A A A E / A A A A A A A 19532 10/ 6 A E A A A A A A A / A A A A A = VIABLE EMBRYO E = NONVIABLE EMBRYO "/" DENOTES POSITION OF CERVIX CLs - CORPORA LUTEA/OVARY 418-013:PAGE B-67 ossooo PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 2) : EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 15 OF PRESUMED GESTATION EMBRYO 8 1 2 3 DOSAGE GROUP III 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 0.4 MG/KG/DAY RAT CLs 19535 8/ 7 A E A A A / E A A A 19537 4/ 8 E / E A A A E 19538 9/ 6 A A A A A A A A / A A A A 19539 9/ 9 A A A A E E A A A / A A A A A A 19540 11/ 7 A A A A A A A A A / A A A A A A A 19542 9/10 A A A A A A A A / A A A A A A A A 19544 12/10 E A A A A A A E / A A A A E A 19547 6/12 A A A A A / A A A A A A A A A A A DOSAGE GROUP IV 1 .6 MG/KG/DAY 19550 9/10 A A A A A A A A A / A A A A A 19553 10/10 A A A A E A A / A A A A A A A 19554 10/ 7 A A A A A A A A A / A A A E A A 19555 NOT PREGNANT 19558 10/ 8 E E E A A AAAAA/AAAAAA 19561a 19563 8/11 A E A A A A/ A A E A A A A 19564 13/13 A A A A A A A/ A E A A A A * VIABLE EMBRYO E * NONVIABLE EMBRYO ./. DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY a. Necropsy and Caesarean-section observations were not recorded Cor rat 19561. A 418-013: PAGE B-68 000SS1 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 3): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 15 OF PRESUMED GESTATION EMBRYO 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP V 3.2 MG/KG/DAY RAT 0 CLs 19565 6/ 7 A A / A A A A 19566 9/ 8 E A A A A A A A / A A A A A 19567 9/ 8 A A A A A A A A / A A A A A 19570 7/10 A A A A A A E / E A A A A A A A A A 19573 5/10 A A A A A / A A E A A A A A A A 19574 10/10 A A A A A A A / A A A A A A 19575 8/11 A A A A A A A / A A E E A A E E A A 19580 11/10 A A A A A A A / A A A A A A A A A VIABLE EMBRYO E NONVIABLE EMBRYO /" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY 418-013: PAGE B-69 0003S2 PROTOCOL 418-013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 4) : EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY RAT # CLs 19503 NO CONFIRMED MATING DATE; DELIVERED ON DAY 66 OF STUDY 19504 10/13 19505 4/16 19506 7/ 8 19507 13/12 19509 7/ 8 19512 4/11 19516 8/ 9 FA E MA E MA MA MA / E MA E MA FA MA E FA MA FA 4.29 4.79 4.76 4.62 4.71 5.10 4.01 4.84 5.21 3.80 4.77 4.83 FA FA MA FA / MA FA MA E FA MA MA FA MA MA MA E FA FA FA 5.04 5.46 5.37 5.41 5.47 4.69 4.15 5.13 5.21 4.81 5.12 5.31 5.36 5.38 4.78 5 .34 4.93 E FA MA FA MA MA / FA FA FA MA FA FA FA FA 5.53 5.93 5.66 5.31 5.43 5.40 5.68 5.64 5.95 4.67 5.87 5.22 5.70 MA FA MA MA MA MA FA FA FA / E FA MA MA FA FA 5.48 5.42 5.34 5.53 5.47 5.64 5.06 5.06 5.28 5.32 5.32 5.49 5.18 5.13 FA FA FA FA MA FA E / MA MA FA FA MA MA 5.03 5.31 4.90 5.51 5.12 5.14 5.44 5.31 5.12 4.98 5.27 5.54 MA FA FA FA / MA FA MA MA MA MA FA FA FA MA S .45 4.87 4.76 5.07 4.78 4.54 4.82 5.29 5.21 4.98 5.30 5.00 4.58 5.47 MA FA FA FA FA FA / MA MA MA FA FA FA MA FA 5.34 4.85 4.87 5.03 5.46 4.92 4.84 5.38 5.06 3.84 5.13 4.91 5.23 4.85 DOSAGE GROUP II 0. 1 MG/KG/DAY 19518 8/ 7 19519 10/ 7 19520 FA E MA FA FA E FA FA / MA MA MA MA MA 4.21 4.70 4.78 4.52 4.53 4.56 4.76 4.70 4.85 4.72 5.07 FA MA FA FA MA FA FA MA MA / FA MA MA MA FA MA MA 4.99 5.31 5.48 5.14 5.63 5.06 4.63 5.31 5.83 5.59 5.16 4.98 4.88 5.22 5.00 5.51 MORIBUND SACRIFICED ON 1DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19521 9 / 6 19523 10/ 4 19524 9/12 19528 6/ 9 19529 8/ 8 FA FA FA MA FA FA MA MA MA / FA FA MA FA MA MA 4.89 4.74 4.62 4.66 4.90 4.81 4.92 4.92 5.28 4.49 4.86 5.20 4.98 5.45 5.42 E FA L FA FA FA FA FA MA MA / MA E FA L 4.80 5.52 5.42 5.23 5.57 4.81 5.58 5.32 5.52 5.62 FA MA MA MA MA FA MA MA / MA MA E MA MA MA FA MA FA 5.16 5.37 5.51 5.64 5.54 5.11 5.70 5.55 5.00 5.49 5.73 5.32 5.56 5.16 5.72 5.00 E MA ,/ MA E E FA MA FA 6.86 6.59 4.83 5.89 5.30 FA MA FA MA E FA FA MA / FA MA FA MA FA MA FA 5.13 5.87 5.63 5.97 5.54 5.64 5.19 5.68 6.11 5.27 6.25 5.71 5.81 5.74 M MALE F 3 FEMALE A - ALIVE E = EARLY RESORPTION L = LATE RESORPTION CLs CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G) . . / . 1DENOTES POSITION OF CERVIX 418-013: PAGE B-70 C00SS3 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 5): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION FETUS # 1 2 DOSAGE GROUP III 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 0.4 MG/KG/DAY RAT CLb 19533 4/12 19534 13/15 19536 7/12 19541 MA MA FA MA / FA FA MA MA MA MA MA FA 5.19 5.23 4.92 5.42 4.80 5.01 5.13 5.09 5.57 5.40 5.35 4.82 FA MA MA MA MA MA FA / FA FA MA MA FA FA MA 4.08 4.80 4.80 4.79 5.20 5.06 5.24 4.78 5.00 5.12 4.87 4.60 5.22 5.32 MA FA FA FA MA FA / PA FA FA FA MA FA FA FA MA FA FA MA 5.57 5.15 5.13 4.97 5.89 5.36 4.88 5.22 5.08 4.93 5.16 4.82 4.74 5.23 4.46 5.12 4.97 5.46 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19543 11/10 19545 E FA MA FA MA MA FA MA MA FA MA / FA 4.83 5.07 4.60 5.10 5.26 4.66 5.08 5.19 4.47 5.51 4.60 MATING NOT CONFIRMED MA MA MA MA MA FA 4.74 5.41 5.48 5.38 4.71 5.17 19546 14/11 MA MA FA 5.04 5.56 4.70 19548 NOT PREGNANT E MA / MA 5.30 4.84 DOSAGE GROUP IV 1.6 MG/KG/DAY 19549 MATING NOT CONFIRMED 19551 MATING NOT CONFIRMED 19552 MATING NOT CONFIRMED 19556 NOT PREGNANT 19557 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19559 NOT PREGNANT 19560 12/ 7 FA FA MA MA MA FA FA FA MA / MA FA FA 5.17 5.13 5.20 5.43 5.44 5.12 5.27 4.70 5.13 5.08 4.85 5.15 O 19562 8/14 MA MA FA FA FA FA MA FA FA FA E MA E MA FA MA FA MA FA MA MA MA 5.50 5.70 4.75 5.27 5.28 4.61 4.84 4.78 4.97 4.74 4.79 4.95 4.93 4.94 4.37 4.75 4.73 5.04 4.77 4.43 M - MALE F - FEMALE A * ALIVE E - EARLY RESORPTION L = LATE RESORPTION */* DENOTES POSITION OF CERVIX CLs CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013:PAGE B-71 00334 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 6): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION FETUS tt 1 2 3 4 S 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 DOSAGE GROUP V 3.2 MG/KG/DAY RAT 8 CLs 1956B 8/11 19569 10/ 6 19571 MA MA MA MA MA MA / E E E 4.12 3.73 4.16 5.19 4.98 4.84 FA MA FA MA FA FA HA MA FA / MA FA FA MA 4.33 4.85 3.43 5.12 4.78 4.33 4.81 4.86 4.66 5.12 5.34 4.61 4.30 DELIVERED ON DAY 21 OF GESTATION 19572 8/ 9 FA- MA FA MA MA 4.51 4.56 4.55 5.02 4.96 19576 MATING NOT CONFIRMED E MA FA / FA 4.31 4.76 4.39 E FA 4.67 E FA MA MA 4.55 4.67 5.15 19577 8/ 5 FA E FA MA FA / 4.98 5.15 5.18 4.87 19578 MATING NOT CONFIRMED 19579 DELIVERED ON DAY 21 OF GESTATION M - MALE F - FEMALE A ALIVE E = EARLY RESORPTION L = LATE RESORPTION "/* DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G). 418-013.PAGE B-72 0003S5 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 21 (PAGE 1) : PLACENTAL WEIGHTS - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION FETUS K 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT * MEAN DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 19503 NO CONFIRMED MATING DATE; DELIVERED ON DAY 66 OF STUDY 19504 19505 19506 19507 19509 19512 19516 0.58 0.44 0.54 0.41 0.53 0.49 0.51 FA E MA E MA MA MA / E MA E MA FA MA E FA MA FA 0.52 0.47 0.59 0.59 0.64 0.45 0.61 0.60 0.69 0.65 0.64 0.50 FA FA MA FA / MA FA MA E FA MA MA FA MA MA MA E FA FA FA 0.53 0.48 0.52 0.47 0.48 0.43 0.40 0.37 0.47 0.38 0.41 0.41 0.48 0.41 0.38 0 .46 0.43 E FA MA FA MA MA / FA FA FA MA FA FA FA FA 0.63 0.48 0.63 0.41 0.61 0.55 0.53 0.52 0.65 0.46 0.53 0.45 0.51 MA FA MA MA MA MA FA FA FA / E FA MA MA FA FA 0.41 0.44 0.38 0.39 0.38 0.44 0.32 0.35 0.38 0.43 0.37 0.44 0.44 0.52 FA FA FA FA MA FA E / MA MA FA FA MA MA 0.46 0.53 0.56 0.53 0.55 0.45 0.57 0.54 0.57 0.54 0.55 0.47 MA FA FA FA / MA FA MA MA MA MA FA FA FA MA 0.56 0.54 0.54 0.54 0.53 0.36 0.46 0.47 0.51 0.40 0.48 0.51 0.47 0.53 MA FA FA FA FA FA / MA MA MA FA FA FA MA FA 0.60 0.46 0.49 0.63 0.63 0.47 0.44 0.48 0.57 0.33 0.61 0.42 0.59 0.42 RAT It MEAN DOSAGE GROUP II 0.1 MG/KG/DAY 19518 19519 19520 0.51 0.45 FA E MA FA FA E FA FA / MA MA MA MA MA 0.49 0.53 0.55 0.54 0.53 0.42 0.66 0.44 0.41 0.46 0.54 FA MA FA FA MA FA FA MA MA / FA MA MA MA FA MA MA 0.43 0.48 0.47 0.47 0.47 0.42 0.38 0.46 0.43 0.44 0.50 0.45 0.55 0.38 0.45 0.49 MORIBUND SACRIFICED ON 1DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19521 19523 19524 19528 19529 0.45 0.53 0.53 0.66 0.62 FA FA FA MA FA FA MA MA MA / FA FA MA FA MA MA 0.44 0.46 0.40 0.44 0.49 0.49 0.48 0.42 0.46 0.37 0.47 0.41 0.48 0.42 0.50 E FA L FA FA FA FA FA MA MA / MA E FA L 0.49 0.55 0.56 0.44 0.55 0.55 0.56 0.49 0.52 0.59 FA MA MA MA MA FA MA MA / MA MA E MA MA MA FA MA FA 0.60 0.52 0.58 0.67 0.47 0.55 0.52 0.58 0.46 0.46 0.67 0.54 0.53 0.40 0.49 0.42 E MA / MA E E FA MA FA 1.08 0.53 0.63 0.52 0.54 FA MA FA MA E FA FA MA / FA MA FA MA FA MA FA 0.93 0.77 0.54 0.63 0.56 0.67 0.69 0.49 0.51 0.50 0.59 0.68 0.53 0.58 M - MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L * LATE RESORPTION "/" 1DENOTES POSITION OF CERVIX CLs CORPORA LUTEA/OVARY PLACENTAL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013: PAGE B-73 C003S6 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 21 (PAGE 2) : PLACENTAL WEIGHTS - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION FETUS ft 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT MEAN DOSAGE GROUP III 0.4 MG/KG/DAY 19533 19534 19536 19541 0.45 0.57 0.46 MA MA FA MA / FA FA MA MA MA MA MA FA 0.52 0.38 0.34 0.44 0.42 0.46 0.40 0.43 0.47 0.58 0.50 0.47 FA MA MA MA MA MA FA / FA FA MA MA FA FA MA 0.49 0.58 0.50 0.51 0.60 0.61 0.62 0.50 0.61 0.53 0.6S 0.61 0.53 0.64 MA FA FA FA MA FA / FA FA FA FA MA FA FA FA MA FA FA MA 0.45 0.48 0.50 0.54 0.53 0.40 0.38 0.54 0.42 0.39 0.49 0.47 0.45 0.47 0.33 0.48 0.46 0.47 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD1 NOT BE DETERMINED 19543 0.41 19545 E FA MA FA MA MA FA MA MA FA MA / FA 0.50 0.39 0.44 0.37 0.49 0.38 0.42 0.43 0.41 0.42 0.28 MATING NOT CONFIRMED E MA MA MA MA MA FA 0.38 0.43 0.43 0.40 0.39 0.42 19546 MA MA FA E MA / MA E E E 0.51 0.44 0.47 0.36 0.49 0.79 19548 NOT PREGNANT RAT ft MEAN DOSAGE GROUP IV 1.6 MG/KG/DAY 19549 MATING NOT CONFIRMED 19551 MATING NOT CONFIRMED 19552 MATING NOT CONFIRMED 19556 NOT PREGNANT 19557 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19559 NOT PREGNANT 19560 19562 0.54 0.44 FA FA MA MA MA FA FA FA MA / MA FA FA 0.48 0.54 0.59 0.67 0.74 0.54 0.50 0.53 0.39 0.56 0.44 0.50 MA MA FA FA FA FA MA FA / FA FA E MA 0.46 0.54 0.43 0.43 0.41 0.45 0.49 0.54 0.53 0.38 0.57 E MA FA MA FA MA FA MA MA MA 0.39 0.34 0.39 0.43 0.45 0.41 0.49 0.48 0.32 M = MALE F FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION /" DENOTES POSITION OF CERVIX CLs = CORPORA LUTEA/OVARY PLACENTAL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-013: PAGE B-74 G003S7 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 21 (PAGE 3) : PLACENTAL WEIGHTS - INDIVIDUAL DATA RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION FETUS ft 1 2 3 4 5 S 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT ft MEAN DOSAGE GROUP V 3.2 MG/KG/DAY 19568 19569 19571 0.62 0.50 MA MA MA MA MA MA / E E E 0.71 0.54 0.76 0.59 0.59 0.54 FA MA FA MA FA FA MA MA FA / MA FA FA MA 0.54 0.45 0.47 0.55 0.51 0.44 0.48 0.50 0.58 0.42 0.56 0.53 0.48 DELIVERED ON DAY 21 OF GESTATION 19572 FA MA FA MA MA 0.42 0.38 0.38 0.39 0.58 0.56 19576 MATING NOT CONFIRMED E MA FA / FA 0.39 0.42 0.38 E FA 0.51 E FA 0.36 19577 FA E FA MA FA / 0.67 0.69 0.68 0.68 0.63 19578 MATING NOT CONFIRMED 19579 DELIVERED ON DAY 21 OF GESTATION M * MALE F FEMALE A ALIVE E = EARLY RESORPTION L * LATE RESORPTION / DENOTES POSITION OF CERVIX CL8 CORPORA LUTEA/OVARY PLACENTAL WEIGHTS WERE RECORDED IN GRAMS (G). 418-013: PAGE B-75 C008S8 APPENDIX C PROTOCOL AND AMENDMENT 418-013:PAGE C-1 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215) 443-8587 PROTOCOL 418-013 SPONSOR'S STUDY NUMBER: T-6295.12 STUDY TITLE: Oral (Gavage) Pharmacokinetic Study of PFOS in Rats PURPOSE: The purpose of this study is to evaluate the pharmacokinetics of PFOS in Fo generation dams and F1 generation fetuses following PFOS treatment of Crl:CDBR VAF/Plus female rats during premating and gestation. TESTING FACILITY: Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Telephone: (215)443-8710 Telefax: (215)443-8587 STUDY DIRECTOR: Raymond G. York, Ph.D., DABT Associate Director of Research SPONSOR: 3M Corporate Toxicology 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 STUDY MONITOR: Marvin T. Case, D.V.M., Ph.D. Telephone: (651)733-5180 Telefax: (651)733-1773 ALTERNATE STUDY MONITOR: Andrew M. Seacat, Ph.D. Telephone: (651)575-3161 Telefax: (651)733-1773 000390 418-013:PAGE C-2 Protocol 418-013 Page 2 REGULATORY CITATIONS: U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Federal Register, September 22, 1994, Vol. 59, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. REGULATORY COMPLIANCE: This study will be conducted in compliance with the Good Laboratory Practice (GLP) regulations cited above. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. The Quality Assurance Unit (QAU) will audit the protocol, the raw data and the report, and will inspect critical phases of the study in accordance with the Standard Operating Procedures of Argus Research Laboratories, Inc. The final report will include a statement signed by the Study Director that the report accurately reflects the raw data obtained during the performance of the study and that all applicable GLP regulations were followed in the conduct of the study. Should significant deviations from GLP regulations occur, each will be described in detail, together with how the deviation might affect the quality or integrity of the study. SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. 000391 418-013: PAGE C-3 Protocol 418-013 Page 3 TEST ARTICLE AND VEHICLE: Identification: Test Article: Name: Physical Description: Lot/Batch Number Specific Gravity: Purity: Expiration Date: PFOS (Synonym: FC-95). Light-colored powder. 217. - 0.6. 98.9%. May 2000. Information on the identity, composition, strength and purity of the test article is on file with the Sponsor. Vehicle: 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Supplier and lot identification of Tween 80 to be documented in the raw data. Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this protocol will be conducted. Safety Precautions: Gloves, mask, appropriate eye protection and a uniform/lab coat are to be worn during formulation preparation and administration. The Material Safety Data Sheet (MSDS) is attached to the protocol (ATTACHMENT 2). Storage: Bulk Test Article: Vehicle Components: Prepared Vehicle: Prepared Formulations: Roomtemperature. Roomtemperature. Roomtemperature. Room temperature. All test article shipments to the Testing Facility should be addressed to the attention of Julian Gulbinski, Manager of Formulations, at the previously cited address and telephone number. 418-013: PAGE C-4 Protocol 418-013 Page 4 Shipments should include information concerning storage conditions and shipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment. FORMULATION: Frequency of Preparation: Formulations (suspensions) will be prepared daily at the Testing Facility. Data verifying the stability of the test article in the vehicle for 48 hours under the conditions of administration are on file with the Sponsor. Detailed preparation procedures are attached to this protocol (ATTACHMENT 3). Adjustment for Puritv: The test article will be considered 100% pure for the purpose of dosage calculations. Testing Facility Reserve Samples: The Sponsor will reserve a sample (1 g) of each lot of the bulk test article used during the course of this study. The Testing Facility will reserve a sample (5 mL) of each lot of the vehicle components used during the course of this study. Samples will be stored under the previously cited conditions. ANALYSES: Samples additional to those described below may be taken if deemed necessary during the course of the study. Bulk Test Article Sampling: No analyses of the bulk test article will be conducted during the course of this study. Information on the stability of the bulk test article is on file with the Sponsor. Analyses of Prepared Formulations: Homogeneity and stability of prepared formulations is on file with the Sponsor. However, records will be maintained to document how the test article formulations were prepared. 00039 418-013:PAGE C-6 Protocol 418-013 Page 6 Body Weight and Aae: Female rats will be ordered to have body weights of 200 g to 225 g each at receipt, at which time they will be expected to be at least 60 days of age. Actual body weights will be recorded the day after receipt and will be documented in the raw data. The weight range will be included in the final report. Sex: Female rats will be given the test article. Male rats of the same source and strain will be used only as breeders and are not considered part of the Test System. Source: Charles River Laboratories, Inc. The rats will be shipped in filtered cartons by air freight and/or truck from Charles River Laboratories, Inc., to the Testing Facility. Identification: Rats are permanently identified using Monel self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats are given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats are assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study. ANIMAL HUSBANDRY: All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratory A n im a te . Housing: Female rats will be individually housed in stainless steel, wire-bottomed cages, except during the cohabitation period and during collection intervals for urine and fecal samples. During cohabitation, each pair of rats will be housed in the male rat's cage. During collection intervals for urine and fecal samples, the female rats will be housed individually in metabolism cages. No nesting materials will be supplied because the female rats will be sacrificed before parturition is expected. 000394 418-013: PAGE C-5 Protocol 418-013 Page 5 Concentration of Test Article Formulations: Concentration of the prepared formulations will be verified during the course of this study. Duplicate samples (2 mL each) will be taken from the first and last preparation on the day prepared. One sample of each set will be shipped for analysis; the remaining samples will be retained at the Testing Facility as backup samples. Backup samples will be stored frozen (-70C or below) and discarded at the Testing Facility upon request of the Sponsor. Shipping Instructions: Samples to be analyzed will be shipped (frozen on dry ice) to: Kris J. Hansen, Ph.D. 3M Environmental Technology and Safety Services 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone: (612) 778-6018 Telefax: (612)778-6176 The recipient will be notified in advance of sample shipment. DISPOSITION: Prepared formulations will be discarded at the Testing Facility. All remaining bulk test article will be returned to the Study Monitor at the previously cited address upon completion of all work with the test article. TEST SYSTEM: Soecies/Strain and Reason for Selection: The Cri:CDBR VAF/Plus (Sprague-Dawley) rat was selected as the Test System because: 1) this strain of rat was used in the reproductive and developmental toxicity studies; 2) historical data and experience exist at the Testing Facility0'3'; and 3) the test article is pharmacologically active in the species and strain. Number: Initial population acclimated: Population selected for study: 120 virgin female rats. 80 mated female rats (16 per dosage group). 418-013:PAGE C-7 Protocol 418-013 Page 7 Room Air. Temperature and Humidity: The animal room is independently supplied with at least ten changes per hour of 100% fresh air that has been passed through 99.97% HEPA filters (Airo Clean room). Room temperature will be maintained at 64F (18C) to 79F (26C) and monitored constantly. Room humidity will also be monitored constantly and maintained at 30% to 70%. Light: An automatically controlled 12-hour light:12-hour dark fluorescent light cycle will be maintained. Each dark period will begin at 1900 hours EST. Diet: Rats will be given Certified Rodent Diet #5002 (PMI Nutrition International) available ad libitum from individual feeders. Water Water will be available ad libitum from individual bottles attached to the cages or from an automatic watering access system. All water will be from a local source and passed through a reverse osmosis membrane before use. Chlorine will be added to the processed water as a bacteriostat; processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. Contaminants: Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the certified diet or the drinking water at levels that would 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: Upon arrival, male and female rats will be assigned to individual housing on the basis of computer-generated random units. After acclimation, virgin female rats will be selected for study on the basis of physical appearance and body weights recorded during acclimation. The female rats will be assigned to dosage groups based on computergenerated (weight-ordered) randomization procedures. 000336 418-013:PAGE C-8 Protocol 418-013 Page 8 Within each dosage group, consecutive order will be used to assign female rats to cohabitation with breeder male rats, one male rat per female rat. The cohabitation period will consist of a maximum of five 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. A table of random units or a computer-generated randomization procedure will be used to select eight female rats per dosage group for Caesarean-section examinations on day 15 of presumed gestation. The remaining female rats in each dosage group will be examined on day 21 of presumed gestation. ADMINISTRATION: Route and Reason for Choice: The oral (gavage) route was selected for use because: 1) this was the route of administration in the developmental and reproductive toxicology studies; and 2) it is one of the possible routes of human exposure. Method and Frequency: Female rats will be given the test article or vehicle once daily beginning 42 days prior to cohabitation through either day 14 of presumed gestation or day 20 of presumed gestation. Dosages will be adjusted daily for body weight changes and given at approximately the same time each day. Rationale for Dosage Selection: Dosages were selected on the basis of a previous study conducted with the test article (Argus Research Laboratories, Inc., Protocol 418-008). Dosage Levels. Concentrations and Volumes: Dosage Group Number of Female Rats Dosage (mg/kg/day) I 16 0 (Vehicle) II 16 0.1 III 16 0.4 IV 16 1.6 V 16 3.2 Concentration (mg/mL) 0 0.02 0.08 0.32 0.64 Dosage Volume (mL/kg) 5 5 5 5 5 Argus Batch Number B-418-013-A(Day.Month. Year) B -4 1 8 -0 1 3-B (Day.Month . Year) B-418 0 1 3-C(Day.Month.Year) B-418-013-D(Day.Month.Year) B-418-013-E(Dey.Month.Year) The test article will be considered 100% pure (dr the purpose of dosage calculations. C00397 418-013: PAGE C-9 Protocol 418-013 Page 9 TESTS. ANALYSES AND MEASUREMENTS: Viability: All Periods: At least twice daily. Clinical Observations and/or General Appearance: Acclimation Period: At least once. Dosage Period: Twice daily. Prior to administration and once approximately one hour postdosage. Postdosage Period: Once prior to sacrifice. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or Study Monitor. Body Weights: Acclimation Period: At least once. Dosage Period: Daily. Sacrifice: Terminal weight. Feed Consumption Values (recorded and tabulated): Acclimation Period: At least once. Dosage Period: Weekly to cohabitation and daily during presumed gestation. Feed consumption values may be recorded more frequently if it is necessary to replenish the feed. These intervals will not be tabulated. 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. 418-013: PAGE C-10 Protocol 418-013 Page 10 Pharmacokinetic Sample Collection: Urine and Fecal Samples: Female rats will be housed individually in metabolism cages for collection of urine and fecal samples for the following intervals: one day prior to initiation of cohabitation to the following morning and days 6 to 7, 14 to 15 and 20 to 21 of presumed gestation. Following each 24-hour collection interval, samples will be collected into centrifuge tubes, placed on dry ice and stored frozen (-70C or below) until shipment for analysis. In the event that a dam begins to deliver before completion of urine and fecal sample collection (day 20 to day 21 of presumed gestation), the dam will be removed from the metabolism cage and placed in a nesting box with sufficient bedding until sacrifice. Blood Samples: Blood samples will be collected from each of the female rats following removal from metabolism caging (prior to administration) on each of the following days: on the day cohabitation is initiated (prior to cohabitation) and on days 7 and 15 of presumed gestation, as well as day 21 of presumed gestation. The time of blood collection will be recorded in the raw data. On ail days of collection except days 15 and 21 of presumed gestation (dams at their terminal collection interval), blood samples (approximately 1 mL each) will be collected from the orbital sinus. If necessary, whole blood may be collected from an alternate site; if so, the alternate site will be documented in the raw data. On day 21 of presumed gestation, blood samples (approximately 4 mL each) will be collected via the inferior vena cava. If blood collection on day 15 of presumed gestation will be a final bleed for the respective dam (i.e., the dam will be Caesarean-sectioned that day), the sample will be approximately 4 mL and will be collected via the inferior vena cava. Blood will be collected and transferred into serum separator tubes. The samples will be spun in a refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. Shipping Instructions: All samples will be maintained frozen (-70 C or below) until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. 000399 418-013: PAGE C-11 Protocol 418-013 Page 11 Caesarean-Sectioning Observations - Dav 15 Presumed Gestation: Eight randomly selected female rats per dosage group will be Caesarean-sectioned on day 15 of presumed gestation. The gravid uterus will be excised and weighed. Placentae that appear abnormal (size, color or shape) will be noted in the raw data. The female rats (pregnant dams) will be examined for number and distribution of: Corpora Lutea. Implantation Sites. Viable and Nonviable Embryos. (A viable embryo is oval or crescent shaped, pink, firm and enclosed in an amniotic sac filled with clear fluid. A nonviable embryo is amorphous, small, pale pink to tan or deep red to black, soft and enclosed in an amniotic sac filled with clear, cloudy or opaque fluid.) Sample Collection: Caps and labeled tubes will be weighed (combined weight, to the nearest .001 gram) before and after retention of pooled embryonic samples for subsequent use in pharmacokinetic analyses. These weights will be documented in the raw data, and copies of these weights will be included with the packing list prior to shipment. Samples of the amniotic fluid of each viable embryo will be collected, pooled (per litter), frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Each viable embryo will be removed from the uterus with the attached placenta, pooled (per litter), frozen and stored (-70C or below) until shipment to the Sponsor. Shipping Instructions: All samples will be maintained frozen (-70C or below) until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. 000300 418-013: PAGE C-12 Protocol 418-013 Page 12 Caesarean-Sectioning Observations - Day 21 Presumed Gestation: All remaining female rats in each dosage group will be Caesarean-sectioned on day 21 of presumed gestation. The gravid uterus will be excised and weighed. Individual placental weights will be recorded. The fetuses will be removed from the uterus, weighed and placed in individual containers. The female rats (pregnant dams) will be examined for number and distribution of: Corpora Ltea. Implantation Sites. [Placentae that appear abnormal (size, color or shape) will be noted in the raw data]. Live and Dead Fetuses. (A live fetus is defined as one that responds to stimuli: a dead fetus is defined as a term fetus that does not respond to stimuli and that is not markedly autolyzed; dead fetuses demonstrating marked to extreme autolysis are considered to be late resorptions.) Early and Late Resorptions. (A conceptus is defined as a late resorption if it is grossly evident that organogenesis has occurred; if this is not the case, the conceptus is defined as an early resorption.) Fetal Observations: Caps and labeled tubes will be weighed (combined weight, to the nearest .001 gram) before and after retention of pooled fetal samples for subsequent use in pharmacokinetic analyses. These weights will be documented in the raw data, and copies of these weights will be included with the packing list prior to shipment. Placental Samples and Amniotic Fluid: Samples of the amniotic fluid (when possible) and the placenta of each fetus will be collected, individually pooled (per litter), frozen and stored (-70C or below) until shipment to the Sponsor for analysis. C00901 418-013: PAGE C-13 Protocol 418-013 Page 13 Gross External Alterations. Sex. Body Weights and Identification: Fetuses will be examined for sex and for gross external alterations. Late resorptions and dead fetuses will be examined for gross external alterations to the extent possible. The individual body weight of each fetus will be recorded. Only body weights of live fetuses will be used to determine litter fetal body weight averages. Representative photographs of fetal gross external alterations will be taken. Blood Samples: Blood samples will be collected from each pup via decapitation, pooled (per litter) and transferred into serum separator tubes. The samples will be spun in a refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (-70 C or below) until shipment to the Sponsor for analysis. Liver Samples: The liver of each fetus will be collected, pooled (per litter), frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Fetal Carcasses: The remaining carcass (and head) of each fetus will be collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Shipping Instructions: All samples will be maintained frozen (-70C or below) until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. METHOD OF SACRIFICE: Rats will be sacrificed by carbon dioxide asphyxiation. Live fetuses will be sacrificed via decapitation. 000902 418-013:PAGE C-14 Protocol 418-013 Page 14 NECROPSY: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation (a table of random units will be used to select one control group rat from which all tissues examined at necropsy will be retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions). Unless specifically cited below, all other tissues will be discarded. Scheduled Sacrifice: On either day 15 of presumed gestation or day 21 of presumed gestation, following the final collection interval for urine and fecal samples and blood sample collection, female rats will be sacrificed, and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. The number and distribution of implantation sites will be recorded. A liver section (right lateral lobe) and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal regions of each dam (left side only) will be collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Uteri of apparently nonpregnant rats will be stained with 10% ammonium sulfide to confirm the absence of implantation sites<5). Rats Found Dead or Moribund: Rats that die or are sacrificed because of moribund condition, abortion or premature delivery will be examined for the cause of death or moribund condition on the day the observation is made. The rats will be examined for gross lesions. A liver section (right lateral lobe) and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal regions of each dam (left side only) will be collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Pregnancy status and uterine contents of female rats will be recorded. Aborted fetuses and/or delivered pups will be examined to the extent possible, using the same methods described for term fetuses. Uteri of apparently nonpregnant rats will be stained with 10% ammonium sulfide to confirm the absence of implantation sites'5*. Shipping Instructions: All samples will be maintained frozen (-70C or below) until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. C00303 418-013: PAGE C-15 Protocol 418-013 Page 15 STATISTICAL EVALUATION. Averages and percentages will be calculated. Litter values will be used where appropriate. Additional procedures and/or analyses may be performed, if deemed appropriate. DATA ACQUISITION. VERIFICATION AND STORAGE: Data will be hand- and/or computer-recorded. Records will be reviewed by the Study Director and/or appropriate management personnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft final report, after which time the Sponsor will be contacted to determine the disposition of these materials. RECORDS TO BE MAINTAINED: Protocol and Amendments. Test Article, Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (if prescribed by Staff Veterinarian). General Comments. Clinical Observations and/or General Appearance. Tissue and Sample Collection, Processing and Shipment. Cap and Labeled Tube Weights. Body Weights. Feed Consumption Values. Caesarean-Sectioning and Fetal 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. 0C0304 418-013:PAGE C-16 Protocol 418-013 Page 16 KEY PERSONNEL: Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Director of Research: Alan M. Hoberman, Ph.D., DABT Associate Director of Research and Study Director Raymond G. York, Ph.D., DABT Director of Laboratory Operations: John F. Barnett, B.S. Director of Study Management: Valerie A. Sharper, M.S. Manager of Animal Operations and Member, Institutional Animal Care and Use Committee: Dena C. Lebo, V.M.D. Director of Operations and Compliance: Barbara J. Patterson, B.A. Consultant, Veterinary Pathology: W. Ray Brown, D.V.M., Ph.D., ACVP FINAL REPORT: A comprehensive draft final report will be prepared on completion of the study and will be finalized following consultation with the Sponsor. The report will include the following: Summary and Conclusion. Experimental Design and Method. Evaluation of Test Results. Appendices: Figures, Summary and Individual Tables Summarizing the Above Data, Protocol and Associated Amendments and Deviations, Study Director's GLP Compliance Statement, Reports of Supporting Data (if appropriate) and QAU Statement. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described in this protocol have been reviewed by the Testing Facility's Institutional Animal Care and Use Committee. All procedures described in this protocol that involve study animals will be conducted in a manner to avoid or minimize discomfort, distress or pain to the animals. The Sponsor's signature below documents the fact that information concerning the necessity for conducting this study and the fact that this is not an unnecessarily duplicative study may be obtained from the Sponsor. No alternative (in vitro) procedures were available for meeting the stated purposes of the study. 000305 418-013:PAGE C-17 Protocol 418-013 Page 17 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, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Cri: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. C00306 PROTOCOL APPROVAL: FOR THE TESTING FACILITY Alan M. Hoberman, Ph.D., DABT Director of Research 418-013:PAGE C-18 Protocol 418-013 Page 18 0 i - ZD6 v Date o q -A jo L f-v z Date J. Patters'^,/B.A. 'Chairperson, Institutional Animal Care and Use Committee FOR THE SPONSOR re . Marvin T. Case, D.V.M., Ph.D. Study Monitor Date Date 000907 418-013:PAGE C-19 ATTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE CC090S ATTACHMENT 1 418-013.PAGE C-20 Protocol 418-013 Page 1 of 2 STUDY SCHEMATIC PHARMACOKINETIC STUDY Start of Dosage Premating Period (42 Days) Cohabitation Period (5 days) Presumed Gestation Period Endof Dosage Caesarean- inninnC Sectioning5 Day15of Day20of Presumed Presumed Gestation Gestation Dosage Period. a. For additional details see "Tests, Analyses and Measurements" section of the protocol. b. Evaluation of corpora lutea, implantation sites and viable and nonviable embryos. c. Fetal evaluations (all fetuses - external examinations). 000909 ATTACHMENT 1 418-013:PAGE C-21 Protocol 418-013 Page 2 of 2 SCHEDULE* 10 NOV 98 16 NOV 98 -2 1 JAN 99 27 DEC 98 PM - 01 JAN 99 AM 28 DEC 98 - 01 JAN 99 12 JAN 9 9 -1 6 JAN 99 18 JAN 99 - 22 JAN 99 26 MAY 99 Animals Arrive - Acclimation Begins. Dosage Period - Female Rats (42 days prior to cohabitation until day 14 or 20 of presumed gestation). Cohabitation Period. Day 0 of Presumed Gestation. Caesarean-Sectioning Period (Day 15 of presumed gestation). Caesarean-Sectioning Period (Day 21 of presumed gestation). Draft Final Report. a. The study initiation date is the day the Study Director signs the protocol. 000910 418-013:PAGE C-22 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 000911 418-013: PAGE C-23 MATERIAL SAFETY DATA SHEET 3M 3M Center St. Paul, Minnesota 55144-1000 1-800-364-3577 or (612) 737-6501 (24 hours) Copyright, 1998, Minnesota Mining and Manufacturing Coapany. All rights reserved. Copying and/or downloading of this inforaation for the purpose of properly utilizing 3M products is allowed provided that: 1) the inforaation is copied in full with no changes unless prior agreeaent is obtained from 3M, and 2) neither the copy nor the original is resold or otherwise distributed with the intention of earning a profit thereon. DIVISION: 3M CHEMICALS TRADE NAME: FC-95 FLU0RAD Brand Fluorocheaical Surfactant 3D NUMBER/U.P.C.: 98-0207-0103-7 00-51135-09054-1 95-0207-0104-5 98-0211-0888-5 00-51135-09362-7 98-0211-3916-1 Z F - 0002-1044-1 ISSUED: January 29, 1998 SUPERSEDES: Noveaber 05, 1997 DOCUMENT: 10-3796-9 00-51135-09055-8 00-51135-02311-2 1. INGREDIENT C.A.S. NO. PERCENT POTASSIUM PERFLUOROALKYL SULFONATE---POTASSIUM PERFLUOROALKYL SULFONATE.... POTASSIUM PERFLUOROALKYL SULFONATE.... POTASSIUM PERFLUOROALKYL SULFONATE.... POTASSIUM PERFLUOROALKYL SULFONATE.... 2795-39-3 3871-99-6 29420-49-3 60270-55-5 3872-25-1 82 3 3 2 1 - 86 -8 -7 -6 -3 2. PHYSICAL DATA BOILING POINT:...... VAPOR PRESSURE:.... VAPOR DENSITY:..... EVAPORATION RAT E : ... SOLUBILITY IN HATER: SPECIFIC GRAVITY:... PERCENT VOLATILE:... P H : ................................ V I S C O S I T Y : .......... MELTING POINT:...... N/A N/A N/A N/A slight ca. 0.6 Hater*l (Bulk) 0% 7 -8 (0.1% Aqueous) N/D N/D APPEARANCE AND ODOR: Light colored, free flowing powder. 000912 Abbreviations: N/D - Not Deterained N/A - Not Applicable CA - Approximately 418-013: PAGE C-24 HBOS: FC-95 FLU0RA0 Brand Fluorochemical Surfactant January 2, 1998 PAGE 2 3. FIRE AND EXPLOSION HAZARD DATA FLASH POINT:.................... None FLAMMABLE LIMITS - LEL:.. HfA FLAMMABLE LIMITS - UEL:.. N/A AUTOIGNITION TEMPERATURE:..... N/A EXTINGUISHING MEDIA: Hater, Carbon dioxide, Dry chemical, Foaa SPECIAL FIRE FIGHTING PROCEDURES: Hear full protective clothing, including helaet, aelf-contained, positive pressure or pressure deeand breathing apparatus, bunker coat and pants, bands around ares, waist and legs, face aask, and protective covering for exposed areas of the head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See Hazardous Decoaposition section for products of coebustion. 4. REACTIVITY DATA STABILITY: Stable INCOMPATIBILITY - MATERIALS/CONDITIONS TO AVOID: Not applicable. HAZARDOUS POLYMERIZATION: Hazardous polyaerization will not occur. HAZARDOUS DECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Sulfur, Hydrogen Fluoride, Toxic Vapors, Gases or Particulates. 5. ENVIRONMENTAL INFORMATION SPILL RESPONSE: Observe precautions froa other sections. Vacuua, use wet sweeping compound or water to avoid dusting. CAUTION! A vacuum cleaner could be an ignition source. Clean up residue with water. Place in an approved aetal container. Seal the container. RECOMMENDED DISPOSAL: Do not release to waterways or sewer. Do not use in products or processes that could result in aquatic concentrations greater than 1/10 of the lowest EC50 or LC50 concentration. Incinerate in an industrial or commercial facility in the presence of a combustible material. Combustion products will include HF. Disposal alternative: Dispose of waste product in a facility permitted to 000313 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-013:PAGE C-25 NSOS: FC-95 FLUORAD Brand Fluoroeheaical Surfactant January 29, 1998 PAGE 3 5. ENVIRONMENTAL INFORMATION (continued) accept chemical waste. ENVIRONMENTAL DATA: 96-Hr. Aquatic Fish LCSQ, Fathaad Minnow(PiaephaIas proaelas)*38 ng/1, B l u e g U l Sunfish(Lepoais aacrochirus)*68 ag/1, Rainbow Trout(Salmo gairdneri)*11 ag/1; 48-Hr. EC50, Daphnia Magna * SO ag/1; COD*.004 g/g; B002D * Nil. REGULATORY INFORMATION: Volatile Organic Coapounds: N/A. VOC Less H20 & Exeapt Solvents: N/A. Since regulations vary, consult applicable regulations or authorities before disposal. U.S. EPA Hazardous Haste Nuaber * None (Not U.S. EPA Hazardous). This product coaplies with the cheaical registration requireaents of TSCA, EINECS, CDSL, AICS, MITI and Korea. EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes 6. SUGGESTED FIRST AID EYE CONTACT: Immediately flush eyes with large aaounts of water for at least 15 minutes. Get immediate medical attention. SKIN CONTACT: Immediately flush skin with large amounts of water. Remove contaminated clothing. If irritation persists, call a physician. contaminated clothing before reuse. Nash INHALATION: If signs/symptoms occur, remove person to fresh air. If signs/symptoms continue, call a physician. IF SHALLOWED: Drink two glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION EYE PROTECTION: Avoid eye contact. Hear vented goggles. O Q 033-4: Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-013:PAGE C-26 MSDS: FC-95 FLUORAD Brand Fluorocheaieal Surfactant January 29, 1998 PAGE 4 7. PRECAUTIONARY INFORMATION (continued) SKIN PROTECTION: Avoid akin contact. Hear appropriate glovaa whan handling this atarial. A pair of glovea Bade froa the following Material(a) are recoBnended: butyl rubber. Uae one or More of the following personal protection iteas as necessary to prevent akin contact: head covering, coveralls. Protective g a m e n t s (other than gloves) should be wade of either of the following Materials: polyethylene/polyvinylidene chloride (Saranex). RECOMMENDED VENTILATION: Use with appropriate local exhaust ventilation. Use in a wellventilatad area. Provide sufficient ventilation to Maintain eaissions below recommended exposure H a l t s . If exhaust ventilation is not adequate, use appropriate reapiratory protection. RESPIRATORY PROTECTION: Avoid breathing of dust. Select one of the following NIOSH approved respirators based on airborne concentration of contaainants and in accordance with OSHA regulations: half-Bask dust and eist respirator, half-Bask supplied air respirator, full-face dust and aist respirator, full-face supplied air respirator. PREVENTION OF ACCIDENTAL INGESTION: Do not eat, drink or snoke when using this product. Wash exposed areas thoroughly with soap and water. Hash hands after handling and before eating. RECOMMENDED STORAGE: Keep container dry. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: Nonflammable. OTHER PRECAUTIONARY INFORMATION: No smoking: Smoking while using this product can result in contaaination of the tobacco and/or saoke and lead to the formation of the hazardous decomposition products aentioned in section 4 of this MSDS. HMIS HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.) EXPOSURE LIMITS INGREDIENT VALUE UNIT TYPE AUTH SKIN* POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 TWA 3M TWA 3M TWA 3M TWA 3M Y Y ._ Y 00031.5 Y Abbreviations: N/D - Not Determined N/A Not Applicable CA - Approximately 418-013:PAGE C-27 MSDS: FC-95 FLUORAD Brand Fluorochaaical Surfactant January 29, 1998 PAGE 5 EXPOSURE LIMITS (continued) INGREDIENT VALUE UNIT TYPE AUTH SKIN* POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 THA 3M Y - SKIN NOTATION: Listed substances Indicated with *Y* under SKIN refer to the potential contribution to the overall expoaure by the cutaneous route including eucous eeabrane and eye, either by airborne or, aore particularly, by direct contact with the substance. Vehicles can alter skin absorption. SOURCE OF EXPOSURE LIMIT DATA: - 3M: 3M Recoaaended Exposure Guidelines 8. HEALTH HAZARD DATA EYE CONTACT: Mild Eye Irritation: signs/syaptoas can include redness, swelling, pain, and tearing. SKIN CONTACT: Mild Skin Irritation (after prolonged or repeated contact): signs/syaptoas can include redness, swelling, and itching. May be absorbed through the skin and persist in the body for an extended tiae. INHALATION: May be haraful if inhaled. May be absorbed by inhalation and persist in the body for an extended tine. Single overexposure, above recoaaended guidelines, nay cause: Irritation (upper respiratory): signs/syaptoas can include soreness of the nose and throat, coughing and sneezing. IF SWALLOWED: Ingestion is not a likely route of exposure to this product. Illness nay result froa a single swallowing of a aoderate quantity of this Material. May be harmful if swallowed. MUTAGENICITY: Mutagenicity assays indicate the product is not autagenic. 000916 Abbreviations: N/D - Not Deterained N/A - Not Applicable CA * Approximately 418-013: PAGE C-28 MSDS: FC-B6 FLUORAD Brand Fluorochamical 8urTactant January 29, 1998 PAGE 6 B. HEALTH HAZARD OATA (continuad) REPRODUCTIVE/DEVELOPHENTAL TOXINS: Not taratogenic in tha rat at oral dosas baloM aatarnally toxic levels. OTHER HEALTH HAZARD INFORMATION: This product is not known to contain any substances regulated under California Proposition 65. A Product Toxicity Suaaary Sheet is available. SECTION CHANGE OATES HEADING SECTION CHANGED SINCE Novenber 05, 1997 ISSUE Abbreviations: N/D - Not Deternined N/A - Not Applicable CA - Approximately The information in this Material Safety Data Sheet (MSDS) is believed to be correct as of the date issued. 3M MAKES NO WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User is responsible for determining whether the 3M product is fit for a particular purpose and suitable for user's eethod of use or application. Given the variety of factors that can affect the use and application of a 3M product, some of which are uniquely within the user's knowledge and control, it is essential that the user evaluate the 3M product to determine whether it is fit for a particular purpose and suitable for user's eethod of use or application. 3M provides information in electronic fora as a service to its customers. Due to the remote possibility that electronic transfer nay have resulted in errors, omissions or alterations in this information, 3M makes no representations as to its completeness or accuracy. In addition, information obtained from a database may not be as current as the information in the MSDS available directly from 3M. P0031-V ^J 418-013: PAGE C-29 ATTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE 000918 418-013:PAGE C-30 ATTACHMENT 3 Protocol 418-013 Version: 418-013 (05 NOV 98) Page 1 of 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE Test Article: PFOS Vehicle: 0.5% Tween 80 in R.O. Deionized Water A. Purpose: The purpose of this procedure is to provide a method for the preparation of dosage suspensions of PFOS and the vehicle for oral administration to rats on Argus Study 418-013. B. General Information: 1. All suspension containers will be labeled and color coded. Each label will specify the protocol number, test article identification, Argus batch number, concentration, dosage level, preparation date, expiration date and storage conditions. 2a. Suspensions will be prepared: X Daily __ Weekly __For____ days of use 2b. Vehicle will be prepared: __ Daily X Weekly __For____ days of use 3. Suspensions will be prepared at a final dosage volume of 5 mL/kg. 4. Safety: X Gloves, lab coat, goggles or safety glasses and faceshield X Dust-Mist Respirator __ Half-Face Respirator __ Full-Face Respirator/Positive Pressure Hood __ Tyvek Suit/Apron 5. Dosage suspensions adjusted for Free base and % Purity. __ Yes X No (Calculations based on 100%) ___ Free Base ___ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol. 000319 418-013:PAGE C-31 ATTACHMENT 3 Protocol 418-013 Version: 418-013 f05 NOV 98) Page 2 of 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE NOTE: Test article will be prepared as a serial dilution from the high dosage to the low dosage. Once the final volumes are achieved, stir bars are to be added to the containers: mixing should occur during sampling and/or administration. C. Preparation of Vehicle 1. Add the required amount of R.O. deionized water to an appropriately labeled container. Heat the water to 50C, 5C, add the required amount of Tween 80 and mix until uniform (See TEST ARTICLE CALCULATIONS). D. Test Article Suspension Preparation: 1. To prepare the 0.64 mg/mL, Group V suspension, add the required amount of test article (See TEST ARTICLE CALCULATIONS) into an appropriately sized, labeled container. QS ad to the required amount with vehicle and heat the mixture to 80C 5C for approximately 30 minutes or until the TA/S dissolves. 2. Once the test article has dissolved; spin while the suspension cools. (Be sure there is a visible vortex, this will achieve the desired emulsion. This may be prepared the day before use.) 3. To prepare the 0.32 mg/mL, Group IV suspension, remove the required amount of stock suspension (Group V) (See TEST ARTICLE CALCULATIONS), QS ad with the vehicle and mix. 4. To prepare the 0.08 mg/mL, Group III suspension, remove the required amount of stock suspension (Group IV) (See TEST ARTICLE CALCULATIONS), QS ad with the vehicle and mix. 5. To prepare the 0.02 mg/mL, Group II suspension, remove the required amount of stock suspension (Group III) (See TEST ARTICLE CALCULATIONS), QS ad with the vehicle and mix. 000920 418-013:PAGE C-32 ATTACHMENT 3 Protocol 418-013 Version: 418-013 35 NOV 981 Page 3 of 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE 6. To prepare the 0 mg/mL, Group I suspension, add required amount of vehicle to an appropriately sized, labeled container (See TEST ARTICLE CALCULATIONS) and mix. Written by: Approved by Clarification:_ No Initials/Date : Ak C - 3 - *31 000921 418-013: PAGE C-33 O P rim edica Argus Research Laboratories. Inc. 905 Sheehy Drive. Building A Horsham. FA 19044 Telephone: {215) 443-8710 Telefax: (215)443-8587 PROTOCOL 418-013 Oral (Gavage) Pharmacokinetic Study of PFOS in Rats SPONSOR'S STUDY NUMBER: T-6295.12 Amendment 1 - 6 January 1999 1. Caesarean-Sectionino Observations - Dav 21 Presumed Gestation (pages 12 and 13 of the protocol) and Necropsy (page 14 of the protocol): At Caesarean-sectioning on day 21 of presumed gestation, lung and liver samples will be collected from three fetuses per litter from five litters per dosage group. These collections will occur after weighing and examination for sex and gross external alterations. One half of the lungs and one lateral lobe of the liver will be individually retained in neutral buffered 10% formalin in scintillation vials. Prior to fixation, several (minimum of two) sections (approximately 1 mm thick) of this half of the lungs and this lobe of the liver will be removed using a scalpel and will be retained in McDowell-Trump's Fixative (stored under refrigeration in scintillation vials) for future evaluation. The other half of the lungs and the other lateral lobe of the liver will be flash frozen in liquid nitrogen and stored in heat-sealable pouches on dry ice. The procedure for collecting and flash freezing these tissues will be available in the raw data. The liver of each of the remaining fetuses, plus any remaining liver from the three selected fetuses, will be collected, pooled (per litter), frozen and stored (-70C or below) until shipment to the Sponsor for analysis as described on page 13 of the protocol. The lung and liver sections in McDowell-Trump's Fixative and the lung and liver samples in neutral buffered formalin will be shipped (on cold packs and ambient conditions, respectively), for possible future evaluation by electron microscopy or light microscopy, respectively, to: 000922 418-013: PAGE C-34 Protocol 418-013 Amendment 1 Page 2 Jeanne deWard Pathology Associates International 4915 D Prospectus Drive Durham, North Carolina 27713 Telephone: (919)544-5257 Telefax: (919)544-3218 The frozen samples will be shipped (on dry ice) to Kris J. Hansen, Ph.D., at the address cited in the protocol for possible future biochemical evaluation. All recipients will be notified in advance of sample shipment. Reason for Change: The Sponsor has requested that these additional samples be retained for continued evaluation of the reasons for reduced pup survival observed in previous studies. T ^ f - Alan M. Hoberman, Ph.D., DABT Director of Research Date Raymond G. York, P h.^^ DABT Associate Director of Research Study Director Date 99 Dena C. Lebo, V.M.D. Date Chairperson, Institutional Animal Care and Use Committee Marvin T. Case, D.V.M., Ph.D. Study Monitor ._ Date 000923 APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 418-013:PAGE D-1 DEVIATION FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY Eighty virgin female rats were randomly assigned to the study and received the test article for forty-two days prior to cohabitation, rather than 120 virgin females assigned to study, dosed, mated and then assigned as the population for the study. Female rats with confirmed date of mating were randomly selected for gestation day 15 (DG 15) Caesarean-sectioning. All remaining rats (with and without confirmed dates of mating) were assigned to DG 21 Caesarean-sectioning. Rats with no confirmed date of mating were Caesarean-sectioned on their estimated DG 21. These deviations did not adversely affect the outcome or interpreation of the study because sufficient rats were available to collect the pharmacokinetic data. All deviat RyQond G. YorRi Rh.D/, DABT Associate Director of Research and Study Director Date APPENDIX E TEMPERATURE AND RELATIVE HUMIDITY REPORTS 000926 ARGUS 418-013:PAGE E-1 Temperature and Relative Humidity Report Location: Room 04 Protocol Number: 418-013 Range of Dates: 10-Nov-1998 15:00 to 23-Nov-1998 15:49 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64*F to 79*F 14 312.75 312 Relative Humidity 30% to 70% 14 312.75 312 Mean ( SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 69.1 ( 2.5) 58.5 (1.7) 76.4 64.4 68.3 58.6 66.8 53.6 312 (100.0) 312 (100.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) Report Generated: 03-Feb-1999 at 13:44 COMMENTS: REVIEWED BY: 0 DATE: Cumulative by Location (v04.01.97) C00927 ARGUS 418-013: PAGE E-2 Temperature and Relative Humidity Report Location: Room 27 Protocol Number: 418-013 Range of Dates: 23-Nov-1998 15:49 to 22-Jan-1999 16:00 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64*F to 79*F 61 1439.74 1442 Relative Humidity 30% to 70% 61 1439.74 1442 Mean ( SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 70.7 72.6 70.8 69.0 1442 0 0 ( 0.7) (100.0) (0.0) (0.0) 54.1 69.8 56.1 14.9 1418 0 24 ( 8.2) (98.3) (0.0) (1.7) Report Generated: 03-Feb-1999 at 13:48 COMMENTS: REVIEWED BY: DATE '. _ 2 h l 2 l Cumulative by Location (v04.01.97) 000923 ARGUS 418-013: PAGE E-3 Relative Humidity Deviations Report Location: Room 27 Protocol Number: 418-013 Range of Dates: 23-Nov-1998 15:49 to 22-Jan-1999 16:00 Humidity Target Range: Species: Rat 30% to 70% Date 04-Jan-1999 05-Jan-1999 06-Jan-1999 06-Jan-1999 Oe-Jan-1999 06-Jan-1999 Oe-Jan-1999 06-Jan-1999 06-Jan-1999 06-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 14-Jan-1999 Time 16:00 23:00 00:00 01:00 02:00 03:00 04:00 05:00 06:00 07:00 11:00 12:00 13:00 14:00 15:00 16:00 00:00 R.H. 29.1 L 26.1 L 23.7 L 23.3 L 20.1 L 18.4 L 17.6 L 16.9 L 14.9 L 27.0 L 27.9 L 29.1 L 26.1 L 28.6 L 26.9 L 28.7 L 29.6 L Date 14-Jan-1999 14-Jan-1999 14-Jan-1999 14-Jan-1999 14-Jan-1999 14-Jan-1999 14-Jan-1999 Time 02:00 03:00 04:00 05:00 06:00 07:00 08:00 R.H. 28.6 L 27.8 L 27.5 L 26.3 L 27.0 L 26.3 L 25.8 L H = Value out of range - High L = Value out of range - Low R.H. = Relative Humidity (%) Report Generated: 03-Feb-1999 at 13:57 i/T h e s e deviations did not adversely affect the outcome or interpretation of the study. __The following deviations) impacted on the outcome of the study as described: Deviations by Location (v04.01.97) APPENDIX F HISTORICAL CONTROL DATA 000930 418-013:PAGE F-1 SUMMARY OF REPRODUCTIVE INDICES CD RAT PERIOD JUNE 1996 - JUNE 1998 NUMBER OF STUDIES 105 NUMBER OF RATS: TESTED PREGNANT FOUND DEAD ABORTED DELIVERED 2233 2065 4 0 0 NUMBER OF RATS PREGNANT AT CAESAREAN-SECTIONING 2052 NUMBER OF RATS WITH SINGLE CONCEPTUS LITTER: LIVE RESORBED ABORTED 6 1 0 MEAN or % % PREGNANT 92.8 AVERAGE # CORPORA LUTEA 16.9 AVERAGE # IMPLANTATIONS 15.3 AVERAGE LITTER SIZE AVERAGE # LIVE FETUSES 14.5 AVERAGE # DEAD FETUSES 0.1 AVERAGE # RESORPTIONS 0.7 AVERAGE # EARLY RESORPTIONS 0.7 AVERAGE # LATE RESORPTIONS 0.0 RANGE/STUDY MEAN or % (64.0-100) (14.0-21.0) (12.9-18.0) (11.8-16.9) (0-1.4) (0-2.0) (0-1.9) (0-0.1) 000931 418-013:PAGE F-2 SUMMARY O F REPRODUCTIVE INDICES CD RAT MEAN or % AVERAGE % DAMS W ITH ANY RESORPTIONS 46.1 AVERAGE % DAMS W ITH ALL CONCEPTUSES RESORBED 0.1 AVERAGE % DAMS W ITH ONE OR MORE LIVE FETUSES 99.8 AVERAGE SEX RATIO, (% MALES/LITTER) 50.2 AVERAGE FETAL BODY WEIGHT (G) 3.48 AVERAGE FOR MALES (G) 3.58 AVERAGE FOR FEMALES (G) 3.39 AVERAGE % DEAD OR RESORBED CONCEPTUSES/LITTER 4.7 RANGE/STUDY MEAN or % (0-87.5) (0-4.5) (95.4-100) (42.8-57.0) (3.10-3.78) (3.17-3.90) (2.98-3.67) (0-12.4) 000332 418-013:PAGE F-3 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT PERIOD JUNE 1996 -JUNE 1998 # STUDIES 124 # RATS TESTED 2668 # RATS PREGNANT 2480* # RATS DIED 8** # RATS ABORTED 0 # RATS DELIVERED 431 # RATS WITH 100% RESORPTION 3 EXTERNAL OBSERVATIONS Red substance around nose and/or mouth White substance present in anterior chamber of eyes Red perivaginal substance MEAN RANGE /STUDY N% N % 2 0.07 0-1 (0-12.5) 1 0.04 0-1 ((M .2) 1 0.04 0-1 (0-4.0) GROSS LESIONS ESOPHAGUS Tom( attributed to an intubation accident) LUNGS Dark red (attributed to an intubation accident) THORACIC CAVITY Filled with light red or cloudy red fluid THYMUS Large AXILLA Mass present Lymph nodes enlarged on left side 1 0.04 0-1 (0-4.0) 1 0.04 0-1 (0-4.0) 2 0.07 0-1 (0-4.0) 1 0.04 0-1 (0-4.0) 1 0.04 0-1 (0-4.0) 1 0.04 0-1 (0-4.2) * Pregnancy status for one dam could not be determined ** One was a monbund sacrifice and one was attributed to an intubation accident C00933 418-013:PAGE F-4 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT GROSS LESIONS LIVER Adhesion between left lateral lobe and left lateral abdominal wall Right lateral and median lobes protruded through diaphragm MEAN RANGE /STUDY N% N % 1 0.04 0-1 (0-4.0) 1 0.04 0-1 (0-4.0) STOMACH/INTESTINE Stomach and intestines distended with gas Cecum contained a black substance Stomach contained a dark red, granular substance 1 0.04 1 0.04 1 0.04 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-4.2) BACK Spine, protrusion of bone on dorsal thoracic portion 1 0.04 0-1 (0-4.0) KIDNEY(S) Pelvis, slight/moderate dilation with or without fluid Pelvis, marked/extreme dilation Mottled Large Small Cortex pitted Left, pelvis contained yellow fluid Bilateral, pelvis contained numerous calculi Right, indentation Pale Left, small; right, enlarged cortex with numerous pinpoint yellow areas and viscous yellow fluid in pelvis 16 0.60 7 0.26 1 0.04 2 0.07 1 0.04 1 0.04 1 0.04 1 0.04 1 0.04 1 0.04 1 0.04 0-2 (0-12.5) 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-3.4) 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-12.5) 0-1 (0-4.0) ADRENAL GLAND(S) Large 1 0.04 0-1 (0 4 .0 ) 000934 418-013:PAGE F-5 SUM M ARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT GROSS LESIONS ABDOMINAL CAVITY Filled with light red fluid MEAN RANGE /STUDY N% N% 1 0.04 0-1 (0-4.0) SPLEEN Two white areas on serosal surface Large 1 0.04 2 0.07 0-1 (0-4.0) 0-2 (0-8.0) BLADDER Wall thick and contained one calculus 1 0.04 0-1 (0-4.0) URETERS Distended with dear fluid 2 0.07 0-1 (0-4.0) VAGINA/CERVIX Cervix contained a dark red, gelatinous substance Cervix contained green, viscous fluid Vagina contained brown, viscous fluid 1 1 1 0.04 0.04 0.04 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-2.1) UTERUS Contained red-brown fluid 1 0.04 Right horn, absent 1 0.04 Right hom, threadlike 1 0.04 Left hom, lumen absent on cervical end; ovarian end, distended with dear fluid 1 0.04 Right hom, dear masses containing gelatinous substance present 1 0.04 Portion of left uterine hom prolapsed 1 0.04 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-2.1) 0-1 (0-4.0) 0-1 (0-4.0) 0-1 (0-4.0) OVARIES Right contained a red, fluid-filled bursal cyst 1 0.04 0-1 (0-4.0) C0Q33S 418-013:PAGE F-6 SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS CD RAT PERIOD JUNE 1996 - JUNE 1998 # STUDIES INCLUDED # LITTERS EXAMINED # LIVE FETUSES EXAMINED 84 1652 23689 ALTERATION HEAD Exencephaly Hematoma Microcephaly L F L F L F EYE(S) Eye bulges depressed Eyelids open Microphthalmia L F L F L F EARS Low set L F SNOUT Short L F TONGUE Absent Protrudes Fused to lower margin of oral opening L F L F L F N% RANGE /STUDY N% 4 0.24 4 0.02 1 0.06 1 0.00 1 0.06 1 0.00 0-1 (0-4-2) 0-1 (0-0.3) 0-1 (0-2.2) 0-1 (0-0.2) 0-1 (0-4.0) 0-1 (0-0.3) 9 0.54 9 0.04 2 0.12 2 0.01 1 0.06 1 0.00 0-2 (0-12.5) 0-2 (0-0.9) 0-1 (0-4.2) 0-1 (0-0.3) 0-1 (0-4.0) 0-1 (0-0.3) 1 0.06 1 0.00 0-1 (0-12.5) 0-1 (0-0.9) 2 0.12 2 0.01 0-1 (0*4.2) 0-1 (0-0.3) 1 0.06 1 0.00 1 0.06 1 0.00 1 0.06 1 0.00 0-1 (0-4.0) 0-1 (0-0.3) 0-1 (0-4.2) 0-1 (0-0.3) 0-1 (0-4.8) 0-1 (0-0.4) L: LITTER INCIDENCE F: FETAL INCIDENCE 000936 418-013: PAGE F-7 SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS CD RAT ALTERATION JAWS Micrognathia Agnathia Cleft BODY Edema Umbilical hernia Spina bifida Hematoma Conjoined twin HINDLIMBS Third limb present ANUS No opening present TAIL Threadlike Agenesis Short Constricted N% RANGE /STUDY N% L 2 0.12 F 2 0.01 L 1 0.06 F 1 0.00 L 1 0.06 F 1 0.00 0-1 (0-4-5) 0-1 (0-0.3) 0-1 (0-12.5) 0-1 (0-0.9) 0-1 (0-4.8) 0-1 (0-0.4) L 2 0.12 F 2 0.01 L 5 0.30 F 5 0.02 L 1 0.06 F 1 0.00 L 2 0.12 F 2 0.01 L 1 0.06 F 1 0.00 0-1 (0-4.3) 0-1 (0-0.3) 0-1 (0-4-3) 0-1 (0-0.3) 0-1 (0-4.2) 0-1 (0-0.3) 0-1 (0-4.0) 0-1 (0-0.3) 0-1 (0-4.0) 0-1 (0-0.3) L 1 0.06 F 1 0.00 0-1 (0-14.3) 0-1 (0-0.9) L 1 0.06 F 1 0.00 0-1 (0-2.6) 0-1 (0-0.2) L 5 0.30 F 5 0.02 L 2 0.12 F 2 0.01 L 1 0.06 F 1 0.00 L 1 0.06 F 1 0.00 0-1 (0-12.5) 0-1 (0-0.9) 0-1 (0-4.5) Qpi (0-0.3) 0-1 (0-2.4) 0-1 (0-0.2) 0-1 (0-2.4) 0-1 (0-0.2) L: LITTER INCIDENCE F: FETAL INCIDENCE e 00937 418-013:PAGE F-8 SUMMARY OF FETAL SOFT TISSUE ALTERATIONS CD RAT PERIOD JUNE 1996 - JUNE 1998 # STUDIES INCLUDED 36 # LITTERS EXAMINED 830 # FETUSES EXAMINED 5753 ALTERATION BRAIN Lateral ventricles, moderate dilation Lateral ventricles, marked dilation Third ventride, marked dilation Lateral and third ventrides, irregularly shaped Irregularly shaped L F L F L F L F L F EYES Microphthalmia L F TONGUE Absent L F JAW Micrognathia L F HEART Septal defect L F VESSELS Innominate, absent L F RANGE/STUDY N% N % 1 0.12 0-1 (0-4.3) 1 0.02 0-1 (0-0.6) 2 0.24 0-1 (0-4.3) 2 0.03 0-1 (0-0.6) 1 0.12 0-1 (0-4.2) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.2) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.2) 1 0.02 0-1 (0-0.6) 3 0.36 0-1 (0-4.3) 3 0.05 0-1 (0-0.7) 1 0.12 0-1 (04.0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 6 0.72 0-2 (0-8.3) 6 0.10 0-2 (0-1.2) L- LITTER INCIDENCE F: FETAL INCIDENCE C00938 418-013:PAGE F-9 SUMMARY O F FETAL SO FT TISSUE ALTERATIONS CD RAT ALTERATION VESSELS (CONT.) Innominate, arises on left Umbilical artery, descends to left of urinary bladder Situs inversus Aorta, descends to right Pulmonary artery, descends to right behind aorta L F L F L F L F L F LUNGS Right apical, cardiac and diaphragmatic lobes appear as one Intermediate lobe, absent L F L F BODY Edema L F ABDOMINAL CAVITY Situs inversus of liver, intestines, stomach, spleen, pancreas and kidneys L F KIDNEY(S) Pelvis, slight dilation L F Pelvis, moderate dilation L F ADRENAL(S) Dark red L F L: LITTER INCIDENCE F: FETAL INCIDENCE RANGE/STUDY N% N % 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 15 1.81 0-2 (0-8.7) 16 0.28 0-2 (0-1.4) 1 0.12 0-1 (0-4.2) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0 4 .0 ) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0 4 .0 ) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0 4 .0 ) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0 4 .0 ) 1 0.02 0-1 (0-0.6) 2 0.24 0-2 (0-8.7) 2 0.03 0-2 (0-1.3) 1 0.12 0-1 (0-4.3) 1 0.02 0-1 (0-0.6) 2 0.24 0-1 (0 4 .3 ) 2 0.03 0-1 (0-0.6) 000939 418-013:PAGE F-10 SUMMARY OF FETAL SKELETAL ALTERATIONS CD RAT PERIOD # STUDIES INCLUDED # LITTERS EXAMINED # FETUSES EXAMINED JUNE 1996 -JUNE 1998 36 816 6151 ALTERATION SKULL Frontal(s): incompletely or not ossified Parietal(s): incompletely or not ossified Nasai(s): short Sphenoid: incompletely ossified Maxillae and Premaxillae: short Skull: incompletely or not ossified VERTEBRAE Thoracic: Centrum, bifid : Centra, unilateral ossification : Centrum, incompletely or not ossified : Centra, fused : Arch, small Lumbar Centrum, bifid : Centrum, incompletely or not ossified : Arches, incompletely or not ossified : Centra, unilateral ossification RANGE/STUDY N% N% L 2 024 F 2 0.03 L 2 0.24 F 2 0.03 L 3 0.37 F 3 0.05 L 1 0.12 F 1 0.02 L 3 0.37 F 3 0.05 L 1 0.12 F 2 0.03 0-1 (0 4 .2 ) 0-1 (0-0.6) 0-1 (0 4 .2 ) 0-1 (0-0.6) 0-1 (0 4 .2 ) 0-1 (0-0.6) 0-1 (0 4 .2 ) 0-1 (0-0.6) 0-1 (0 4 .2 ) 0-1 (0-0.6) 0-1 (0 4 .2 ) 0-2 (0-1-2) L 67 8.21 F 74 1.20 L 6 0.74 F 6 0.10 L 3 0.37 F 4 0.06 L 1 0.12 F 1 0.02 L 1 0.12 F 1 0.02 L 1 0.12 F 1 0.02 L 2 0.24 F 3 0.05 L 12 1.47 F 17 0.28 L 2 0.24 F 2 0.03 0-5 (0-20.8) 0-6 (0-3.4) 0-2 (0-8.0) 0-2 (0-1.0) 0-2 (0-8.3) 0-3 (0-1.8) 0-1 (0 4 .2 ) 0-1 (0-0.5) 0-1 (0 4 .0 ) 0-1 (0-0.6) 0-1 (0 4 .3 ) 0-1 (0-0.6) 0-2 (0-8.3) 0-3 (0-1.8) 0-3 (0-12.0) 0 4 (0-2.1) 0-1 (0 4 .2 ) 0-1 (0-0.6) L: LITTER INCIDENCE F: FETAL INCIDENCE 000940 418-013:PAGE F-11 SUMMARY OF FETAL SKELETAL ALTERATIONS CD RAT ALTERATION RIBS Cervical Rib(s) present One or more, wavy One or more, incompletely ossified (hypoplastic), or not ossified Extra ribs, 14th, unilateral Extra ribs, 14th, bilateral RANGE/STUDY N% N % L 23 2.82 F 26 0.42 L 32 3.92 F 52 0.84 0-4 (0-17.4) 0-5 (0-2.9) 0-4 (0-33.3) 0-8 (0-5.1) L 22 2.70 F 36 0.58 L 4 0.49 F 4 0.06 L 1 0.12 F 1 0.02 0-2 (0-9.1) 0-5 (0-2.6) 0 4 (0-16.0) 0 4 (02.2) 01 (04.0) 0 1 (0-0.6) STERNEBRAE One o r more incompletely ossified or not ossified Fused Asymmetric L 109 13.36 F 150 2.44 L 1 0.12 F 1 0.02 L 1 0.12 F 1 0.02 0 9 (036.0) 0 1 2 (0-6.7) 01 (04.2) 0 1 (0-0.6) O I (04.2) 0 1 (0-0.5) SCAPULAE Bent L 1 0.12 F 1 0.02 01 (04.2) 0 1 (0-0.5) PELVIS Pubis(es) and/or Ischium(a): incompletely or not ossified Pubis(es): incompletely ossified Pubis(es): not ossified Ischium(a): incompletely or not ossified L 116 14.22 F 180 2.93 L 96 11.76 F 147 2.39 L 3 0.37 F 3 0.05 L 39 4.78 F 58 0.94 0 7 (030.4) 016 (09.5) 0 7 (030.4) 0-16 (0 9 .5 ) 01 (04.2) 01 (00.6) 0 4 (016.7) 0 9 (0-4.7) FORELIMB(S) Radius and Ulna: Bent L 1 0.12 F 1 0.02 0 1 (04.2) 0 1 (00.6) L: LITTER INCIDENCE F: FETAL INCIDENCE 000941 418-013:PAGE F-12 SUMMARY OF FETAL OSSIFICATION SITES . SKELETAL AVERAGES CD RAT (CAESAREAN-SECTIONED DAY 20 GESTATION) PERIOD: JUNE 1996 -JUNE 1998 # STUDIES INCLUDED 35 # LITTERS EXAMINED 793 # FETUSES EXAMINED 5961 SKELETAL AVERAGES HYOID VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS (pairs) STERNUM MANUBRIUM STERNAL CENTERS XIPHOID FOREPAWS (Calculated as average per limb) CARPALS METACARPALS DIGITS PHALANGES HINDPAWS (Calculated as average per limb) TARSALS METATARSALS DIGITS PHALANGES FETUS/LITTER MEAN RANGE/STUDY 0.86 7.00 13.04 5.95 3.00 4.82 13.03 (0.69-0.98) _ (13.00-13.15) (5.85-5.99) (2.96-3.01) (4.35-5.16) (13.00-13.09) 1.00 (0.98-1.01) 3.61 (3.26-3.75) 0.99 (0.94-1.00) 0.00 3.54 5.00 5.04 (3.33-3.74) -- (4.90-5.27) 0.00 3.99 5.00 4.98 (3.93-4.03) -- (4.82-5.08) 000942 418-013:PAGE F-13 FETAL OSSIFICATION SITES SKELETAL AVERAGES CD RAT (CAESAREAN-SECTIONED DAY 21 GESTATION) STUDY# # LITTERS EXAMINED # FETUSES EXAMINED SKELETAL AVERAGES HYOID VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS (pairs) STERNUM MANUBRIUM STERNAL CENTERS XIPHOID FOREPAWS (Calculated as average per limb) CARPALS METACARPALS DIGITS PHALANGES HINDPAWS (Calculated as average per limb) TARSALS METATARSALS DIGITS PHALANGES 580 23 190 0.96 7.00 13.06 5.94 3.00 7.23 13.05 1.00 3.97 1.00 0.00 3.99 5.00 7.53 0.02 4.62 5.00 5.78 000943 APPENDIX G STATEMENT OF THE STUDY DIRECTOR 000944 OPrimedica 418-013:PAGE G-1 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215)443-8587 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.12 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 Welfare (MHW) Good Laboratory Practice Standard fo r Safety Studies on Drugs? and the European Economic Community (EEC) Council decision on 28 July 1989 on the acceptance by the European Economic Community o f an OECD decision/recommendation on compliance with principles o f good laboratory practice0occurred that affected the quality or integrity of the study. ^-ym ond G. Y Associate Dired and Study Direct ,D,, DABT Date Research a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1988). Good Laboratory Practice Standard for Safety Studies on Drugs, M HW Ordinance Number 21, March 26, 1997. c. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community o f an OECD decision/recommendation on compliance with principles o f good laboratory practice. Official Journal of the European Communities: Legislation. 32(No. L 315; 28 October): 1-17. 000945 APPENDIX H QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT 000946 O P rim edica 418-013:PAGE H-1 Argus Research Laboratories. Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215)443-8587 QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT Study Director: Raymond G. York, Ph.D., DABT Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Protocol 418-013: Oral (Gavage) Pharmacokinetic Study of PFOS in Rats Sponsor's Study Number T-6295.12 The draft protocol for this study was audited for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and 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 on 18 OCT 98. Critical phases of this study were inspected six times; study information and raw data were audited twice (see tables 1 and 2 for dates and phases/data) The draft final report and the raw data for this study were compared and audited for accuracy, for adherence to protocol requirements, and for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and 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 between 25 APR 99 and 26 MAY 99, and for revisions requested by the Sponsor 17 JUN and 18 JUN 99 and for finalization on 24 JUN 99. 000947 418-013:PAGE H-2 This study was conducted according to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and 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. Nancy J. Gongliewski Date j#J_isa A. Zaborowski, B.S. Date Quality Assurance Manager ( j Senior Quality Assurance Associate and Principal Auditor 000948 TABLE 1 CRITICAL PHASES INSPECTED 418-013:PAGE H-3 Test Article Preparation Date of inspection: 19 NOV 98 Date results reported to the Study Director and Management: 20 NOV 98 Test Article Administration - Gavaae Date of inspection: 19 NOV 98 Date results reported to the Study Director and Management: 20 NOV 98 Cohabitation Date of Inspection: 28 DEC 98 Date results reported to the Study Director and Management: 31 DEC 98 Blood Collection Date of inspection: 04 JAN 99 Date results reported to the Study Director and Management: 04 JAN 99 Urine and Fecal Collection Date of inspection: 04 JAN 99 Date results reported to the Study Director and Management: 04 JAN 99 Caesarean-Sectioning Date of inspection: 13 JAN 99 Date results reported to the Study Director and Management: 19 JAN 99 00949 418-013:PAGE H-4 TABLE 2 RAW DATA AUDIT(S) The following study information and raw data were audited from 18 MAR 99 to 25 MAR 99: Protocol. Protocol amendment. List of personnel and computer operator codes. Error codes and codes for clinical sign observations. Animal receipt, randomization, physical examination and acclimation. In-life transaction record. Feed consumption. Cohabitation. Caesarean-sectioning. Maternal gross observations. Fetal gross observations. Necropsy. Gravid uterine weights. Placental weights. Tissue packing lists. Male breeder colony records. General comments. Study maintenance records. Temperature and relative humidity reports. Feed and water analyses. Edit requests. Randomizations. Dosage volumes. Deviations. Data review pages. Blood and liver collection data and packing lists. Maternal packing lists for liver/glands. Fetal liver/lung tissue packing lists. Pooled liver, placenta, liver and lung packing lists. Fecal/urine collection data and packing lists. Embryo/placenta collection and packing lists. Fetal carcass packing lists. Amniotic fluid collection and packing lists. The results of this audit were reported to the Study Director and Management on 26 MAR 99. 00950 418-013:PAGE H-5 The following study information and raw data were audited on 25 MAR 99: Vehicle/Control article receipt, preparation and use. Test article/substance receipt, preparation and use. Test article/substance packing lists. Test article/substance analysis. The results of this audit were reported to the Study Director and Management on 29 MAR 99. 000951
Contact UsLoginSign Up
CUNY - The Graduate CenterColumbia University Mailman School of Public Health - Sociomedical Sciences