Document QJ2jE242LOJEqBQygVXEQ7oG7

COVArfiCE^ THE DEVELOPMENT SERVICES COMPANY fiR AS. 6 -- 13 S'S' Sponsor: 3M St. Paul, Minnesota FINAL REPORT Study Title: 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Author: Peter J. Thomford, PhD Study Completion Date: April 9, 2002 Performing Laboratory: Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704-2595 Laboratory Project Identification. Covance 6329-222 Sponsor Project Identification: 3M Study No. T-6295.6 Page 1 o f 235 000137 Covance 6329-222 3M T-6295.6 COMPLIANCE STATEMENT 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys All aspects o f this study were in accordance with the Environmental Protection Agency Good Laboratory Practice (GLP) Standards, 40 CFR 792. However, during the timeframe of March 10, 1998, through April 16, 1998, prestudy procedures such as general husbandry, health screening activities, and testing for assignment to study were conducted. The protocol was finalized on April 17, 1998. While the performance of these prestudy events without a final protocol represents a GLP deviation, they did not affect either the outcome or the interpretation of the data. Study rector / Covance Laboratories Inc. Andrew M. Seacat, PhD Study Monitor 3M Date 000138 2 Covance 6329-222 3M T-6295.6 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit o f Covance Laboratories Inc. The following inspections were conducted and findings reported to the study director (SD) and associated management. Inspection Dates From To 04/21/98 04/21/98 05/20/98 05/20/98 05/20/98 05/20/98 05/21/98 05/21/98 06/01/98 06/01/98 07/08/98 07/13/98 08/14/98 08/14/98 08/10/98 08/18/98 06/04/99 06/04/99 03/27/02 03/27/02 03/27/02 03/27/02 Phase Protocol Review Protocol Amendment Review Protocol Amendment Review Clinical Laboratory Inspection Postlife Data Review Protocol Amendment Review Report Review Protocol Amendment Review Protocol Amendment Review Report Review Date Reported to Study Director and Study Director Management 04/21/98 05/20/98 05/20/98 05/21/98 06/01/98 07/13/98 08/14/98 08/18/98 06/04/99 03/27/02 03/27/02 Representative Quality Assurance Unit ( y I A ril 3 0 0 $ Date ' 000139 3 STUDY IDENTIFICATION Covance 6329-222 3MT-6295.6 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Test Material Sponsor Study Monitor Alternate Study Monitor Study Location Study Director Study Timetable Study Initiation Date In-Life (Experimental) Start Date In-Life Termination Date Experimental Termination Date Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) 3M Toxicology Services Building 220-2E-02, 3M Center St. Paul, Minnesota 55144-1000 Andrew M. Seacat, PhD 3M 651.575.3161 Marvin T. Case, DVM, PhD 3M 651.733.5180 Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704-2595 Peter J. Thomford, PhD Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704-2595 608.241.7207 April 17, 1998 April 23, 1998 May 22, 1998 April 8, 2002 000140 4 KEY PERSONNEL Covance 6329-222 3M T-6295.6 Study Director Study Toxicologist Study Coordinator Supervisor, Large Animal Toxicology Supervisor, Dose Formulation Supervisor, Laboratory Animal Medicine Clinical Pathologist Supervisor, Clinical Pathology Anatomical Pathologist Anatomical Pathologist Supervisor, Anatomical Pathology Peter J. Thomford, PhD Dale Aldridge, BS Patricia K. McKee Pesik, BS, LAT Meechelle Bordeaux, LAT Dixie Bushee, BS, LATG Donna J. Clemons, DVM, MS Diplomate, ACLAM Robert L. Hall, DVM, PhD Diplomate, ACVP (Clinical Pathology) Ronald Markevitch, BS, MT (ASCP) Dawn G. Goodman, VMD Diplomate, ACVP Director of Pathology Thomas E. Palmer, PhD Kimberly W. Durland, BS, HT 00141 5 Covance 6329-222 3M T-6295.6 CONTENTS Page ABSTRACT................................................................................................................................ 10 PURPOSE................................................................................................................................... 12 REGULATORY COMPLIANCE.............................................................................................12 TEST MATERIAL, VEHICLE, AND SOLVENT................................................................ 12 Test M aterial.......................................................................................................................... 12 V ehicle.................................................................................................................................... 12 Solvent.................................................................................................................................... 13 Reserve (Archive) Samples...................................................................................................13 D is p o s itio n .............................................................................................................................. 13 TEST SYSTEM ......................................................................................................................... 13 Test Animal.............................................................................................................................13 Identification...........................................................................................................................13 A c c lim a tio n ............................................................................................................................. 13 Housing and Maintenance..................................................................................................... 14 Justification............................................................................................................................. 15 P R O C E D U R E S .......................................................................................................................... 15 Group Designations and Dose Levels..................................................................................15 Dose Preparation.................................................................................................................... 15 Dose Analyses.........................................................................................................................17 Method o f A dm inistration.................................................................................................... 17 Clinical Observations.............................................................................................................17 Body Weights..........................................................................................................................18 Rectal Body Temperatures.................................................................................................... 18 Blood Hormone Determination.............................................................................................18 Serum PFOS Level Determination...................................................................................... 18 Clinical Pathology.................................................................................................................. 19 Additional Blood Collection.................................................................................................19 N ecropsy.................................................................................................................................20 Palmitoyl CoA Oxidase Determinations............................................................................. 20 Cell Proliferation Evaluation................................................................................................20 Liver PFOS Determ ination...................................................................................................20 Tissue Preservation................................................................................................................21 Histopathology....................................................................................................................... 21 Statistical A nalyses................................................................................................................22 RECORD RETENTION........................................................................................................... 22 6 000142 Covance 6329-222 3M T-6295.6 CONTENTS (Continued) Page RESULTS................................................................................................................................... 23 Dose Analyses........................................................................................................................ 23 Clinical Observations............................................................................................................ 23 Body Weights......................................................................................................................... 23 Food Consumption.................................................................................................................24 Rectal Body Temperatures....................................................................................................24 Blood Hormone Analyses.....................................................................................................24 Clinical Pathology..................................................................................................................25 Cell Proliferation Evaluation............................................................................................... 25 Anatomical Pathology........................................................................................................... 25 CONCLUSIONS........................................................................................................................ 25 SIGNATURES........................................................................................................................... 26 REFERENCES.......................................................................................................................... 27 PATHOLOGY REPORT.......................................................................................................... 28 COMMENTS ON THE D A T A ............................................................................................... 31 CODES, ABBREVIATIONS, AND U N ITS..........................................................................32 General Codes and A bbreviations.......................................................................................33 Codes for Clinical Pathology............................................................................................... 34 Abbreviations and Units for Clinical Hematology............................................................36 Abbreviations and Units for Clinical C hem istry...............................................................38 Codes for Anatomical Pathology.........................................................................................40 TABLES 1 Summary o f Clinical Observations A.M./W eekly......................................................... 41 2 Summary o f Clinical Observations 30 Minutes Postdose............................................. 42 3 Summary o f Clinical Observations 60 Minutes Postdose............................................. 43 4 Summary o f Clinical Observations 90 Minutes Postdose............................................. 44 5 Summary o f Clinical Observations Unscheduled.......................................................... 45 6 Summary o f Body Weight Data (k g )............................................................................... 46 7 Summary o f Rectal Body Temperature Data (C )......................................................... 49 8 Summary o f Clinical Hematology Data - Day -7 ............................................................51 9 Summary o f Clinical Hematology Data - Day 2 9 ...........................................................55 10 Summary o f Clinical Chemistry Data - Day -7.............................................................59 11 Summary o f Clinical Chemistry Data - Day 2 ..............................................................65 12 Summary o f Clinical Chemistry Data - Day 7 ..............................................................71 13 Summary o f Clinical Chemistry Data - Day 14............................................................77 7 000143 Covance 6329-222 3M T-6295.6 CONTENTS (Continued) Page TABLES 14 Summary of Clinical Chemistry Data - Day 2 9 ............................................................83 15 Summary of Clinical Chemistry Data - Day 3 0 ............................................................89 16 Incidence of Macroscopic Observations........................................................................91 17 Incidence o f Microscopic Observations.........................................................................92 APPENDIX 1..............................................................................................................................94 Protocol Deviations................................................................................................................95 Protocol................................................................................................................................... 97 Protocol Amendment No. 1 ................................................................................................115 Protocol Amendment No. 2 ................................................................................................117 Protocol Amendment No. 3 ................................................................................................120 Protocol Amendment No. 4 ................................................................................................123 Material Safety Data S heet.................................................................................................125 APPENDIX 2 ............................................................................................................................131 Individual Animal Fate Data............................................................................................ 132 Individual Clinical Observations......................................................................................133 Individual Rectal Body Temperature Data (C)............................................................... 140 APPENDIX 3 ............................................................................................................................142 Individual Body Weight Data (kg)..................................................................... APPENDIX 4 ............................................................................................................................145 Individual Clinical Hematology D ata.............................................................................. 146 Individual Clinical Chemistry D a ta ................................................................................. 158 APPENDIX 5 ............................................................................................................................ 190 Individual Animal Pathology D a ta ....................................................................................191 APPENDIX 6 ............................................................................................................................203 Quality Assurance Statement - AniLytics.........................................................................204 Summary and Individual Hormone Analyses D ata..........................................................205 Figure 1 - Mean Estradiol Data (pg/mL) - M ales.......................................................... 209 Figure 2 - Mean Estradiol Data (pg/mL) - Females....................................................... 210 Figure 3 - Mean Triiodothyronine Data (ng/dL) - M ales.............................................. 211 Figure 4 - Mean Triiodothyronine Data (ng/dL) - Fem ales.......................................... 212 APPENDIX 7 ............................................................................................................................213 Dose Confirmation Analysis Report..................................................................................214 Compound Stability R eport................................................................................................219 143 8 000144 Covance 6329-222 3MT-6295.6 CONTENTS (Continued) Page Certificate o f A nalysis........................................................................................................ 221 Quality Assurance Statement - 3 M ....................................................................................226 APPENDIX 8 ............................................................................................................................227 Cell Proliferation Report.....................................................................................................228 000145 9 ABSTRACT Covance 6329-222 3M T-6295.6 The purpose of this study was to provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study and to assess the effect of the test material, Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295), on critical enzyme levels, hormones, and other selected biochemical parameters. Male and female cynomolgus monkeys were assigned to three groups (two animals/sex in Group 1, three animals/sex in Group 2, and one animal/sex in Group 3). Animals in Groups 2 and 3 received gelatin capsules containing 0.02 and 2.0 PFOS/kg of body weight/day (mg/kg/day), respectively, triturated with lactose (1:999 and 1:9, w:w, respectively). Animals in Group 1 received gelatin capsules containing lactose only. The total material dose level for each group was 20.0 mg/kg/day. Food was provided once or twice daily. Water was provided ad libitum. The animals were observed twice daily (a.m. and p.m.) for mortality and moribundity. At least once daily, animals were examined for abnormalities and signs o f toxicity, and food consumption was assessed qualitatively. In addition, animals were observed approximately 30, 60, and 90 minutes postdose for signs o f poor health or abnormal behavior. Body weight data were collected twice weekly beginning 2 weeks before initiation o f treatment, on Day -1, on the first day o f treatment, and twice weekly thereafter. Rectal body temperatures were recorded twice weekly beginning 2 weeks before initiation o f treatment. Blood samples for hormone analyses [estradiol, estrone, estriol, thyroid stimulating hormone, triiodothyronine (T3), and thyroxin (T4)] were collected before initiation o f treatment and predose on Day 29. Serum was harvested, and the samples were sent to Ani Lytics Inc., for analyses. Blood samples for serum PFOS concentration analyses were collected before initiation of treatment and predose on Days 2, 3 (approximately 24 hours after the first and second dose, respectively), 7, 14, and 29. Serum was harvested, and samples were sent to the Sponsor for analyses. Blood samples were collected for hematology and clinical chemistry tests before initiation of treatment and on Day 29. In addition, blood for clinical chemistry tests was collected predose on Days 2 ,7 , and 14. Additional blood was collected at the time o f exsanguination, and samples were shipped to the Sponsor for possible future analysis. On Day 30, the animals were anesthetized, weighed, exsanguinated, and necropsied. At necropsy, macroscopic observations were recorded, and selected tissues were collected 10 000146 Covance 6329-222 3M T-6295.6 and preserved. In addition, a sample o f liver was collected from each animal for palmitoyl CoA oxidase activity analyses. Representative samples o f liver, testes, and pancreas were collected from each animal and sent to Pathology Associates International for proliferation cell nuclear antigen evaluation. A sample o f liver was collected from each animal and sent to the Sponsor for PFOS concentration analyses. Microscopic examinations were done on the adrenals, eye, femoral bone marrow, lung, liver, kidney, pancreas, spleen, testes, and thymus from each animal. All animals survived to the scheduled sacrifice. There were no test material-related effects on clinical observations, body weights, food consumption, rectal body temperatures, clinical pathology, or macroscopic or microscopic findings. There were no apparent test material-related effects on estrone, estriol, thyroid stimulating hormone, or thyroxin. There were no apparent test material-related effects on estradiol or triiodothyronine at 0.02 mg/kg/day. Given the low number of animals in this study, it is not possible to determine conclusively whether there was a test material-related effect on estradiol or triiodothyronine. Estradiol levels for the male and female given 2.0 mg/kg/day were numerically lower than those of the control animals on Day 29 and their respective prestudy values. Triiodothyronine levels for the male and female given 2.0 mg/kg/day were numerically lower than those of the control animals on Day 29; however, their prestudy values were also considerably lower than the control values. Triiodothyronine levels for animals given 0 or 0.02 mg/kg/day were numerically higher on Day 29 compared with their prestudy levels while triiodothyronine levels for animals given 2.0 mg/kg/day were numerically lower compared with their prestudy levels. Cell proliferation, as determined by measuring the proliferating index, was not increased in the liver, testes, or pancreas of monkeys. In cynomolgus monkeys treated with 0, 0.02, or 2.0 mg PFOS/kg/day orally for at least 4 weeks, there were no apparent test material-related effects on clinical observations, body weights, food consumption, rectal body temperatures, or clinical or anatomical pathology findings. The one male and one female treated with 2.0 mg PFOS/kg/day appeared to have lower levels o f estradiol on Day 29. 000147 n PURPOSE Covance 6329-222 3M T-6295.6 The purpose o f this study was to provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study and to assess the effect o f the test material, Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295), on critical enzyme levels, hormones, and other selected biochemical parameters. REGULATORY COMPLIANCE All aspects o f this study were done in accordance with the Environmental Protection Agency Good Laboratory Practice Standards, 40 CFR 792. TEST MATERIAL, VEHICLE, AND SOLVENT Test Material The test material, Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295), Lot No. 217, is a white to off-white powder and is 86.9% pure. It was received at Covance on September 4, 1997. The test material was stored at room temperature. Information on synthesis methods, stability, purity, composition, or other characteristics that define the test material is on file with the Sponsor. The Certificate o f Analysis is in Appendix 7. Vehicle The vehicle was lactose (Spectrum, New Brunswick, New Jersey), Lot No. NN0192 (expiration date February 13,1999). It was received at Covance on March 30,1998. The vehicle was stored at room temperature. Information on synthesis methods, purity, composition, or other characteristics that define the vehicle is on file with the manufacturer. Q00148 Covance 6329-222 3M T-6295.6 Solvent The solvent was acetone, USP (Spectrum, Gardena, California), Lot No. LH0253 (expiration date June 2000), and is 99.8% pure. It was received at Covance on June 23, 1997. The vehicle was stored at room temperature. Information on synthesis methods, purity, composition, or other characteristics that define the solvent is on file with the manufacturer. Reserve (Archive) Samples A reserve sample (10 g each) o f each lot o f the test material, vehicle, and each test material/lactose trituration was taken before initiation of treatment and stored at room temperature. These samples were transferred to the Sponsor after completion o f the in-life phase. Disposition Remaining test material will be retained at Covance for use in possible future studies. TEST SYSTEM Test Animal Young adult to adult cynomolgus monkeys were obtained from Covance Research Products Inc. (Alice, Texas), on March 10, 1998. The animals weighed 2.0 to 2.5 kg at initiation o f treatment. Identification Each animal was assigned a permanent number upon arrival and identified with collar tag before initiation o f treatment. All data for an animal are recorded under this number. Acclimation Seven males and seven females were received on March 10, 1998, transferred from the Covance in-house stock colony on March 16, 1998, and acclimated in Animal Room 2015 for 38 days before initiation o f treatment. In general, animals in this 13 000149 Covance 6329-222 3MT-6295.6 shipment appeared healthy. During acclimation, the animals were examined for abnormalities indicative of health problems. In addition, three tuberculosis tests, a physical examination, and a fecal flotation for parasites were done on each animal. The physical examination done after the animals were received revealed no major abnormalities. Results o f the tuberculosis tests were negative for all animals; the results are recorded in the data. The results of the fecal flotation tests indicated the animals were negative for parasite ova. One male and one female were eliminated from study consideration based on clinical pathology findings or abnormal fecal observations during acclimation. There were six males and six females in the randomization for assignment to groups (two animals/sex in Group 1, three animals/sex in Group 2, and one animal/sex in Group 3). Animals not selected for the study were transferred to the Covance stock colony. Housing and Maintenance Animal Room 2015 was used for this study. Environmental controls for the animal room were set to maintain 18 to 29C, a relative humidity o f 30 to 70%, and a 12-hour light/12-hour dark cycle. Variations from these conditions are documented in the data and are considered to have had no effect on the outcome o f the study. The animals were housed individually in stainless steel cages. Certified primate diet (#8726C, Harlan Teklad) was provided once or twice daily. The lot numbers are recorded in the data. The diet is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Results o f specified nutrient and contaminant analyses are on file with Covance-Madison. Fruits or additional supplements were provided, but did not require analysis. Water was provided ad libitum. Samples of the water are analyzed for specified microorganisms and environmental contaminants. The results are on file with Covance-Madison. There were no known contaminants in the diet or water at levels that would have interfered with this study. 000150 14 Covance 6329-222 3M T-6295.6 Justification PFOS is a known hepatic peroxisome proliferator (PP) in the rat. When exposed to PP, nonhuman primates (such as the cynomolgus monkey) respond similarly to humans (i.e., low to no hepatic response) and therefore are an appropriate human surrogate species. PROCEDURES This study was conducted in accordance with the Protocol dated April 17, 1998, and Protocol Amendment Nos. 1 through 4. The protocol, protocol amendments, and protocol deviations are in Appendix 1. Group Designations and Dose Levels Selection o f animals for the study was based on clinical observations, clinical pathology, and other data as appropriate. Animals were assigned to treatment groups using a computerized blocking procedure designed to achieve body weight balance with respect to treatment group. Dose Level Total Material Dose Number of Animals Group (mg/kg/day) Level (mg/kg/day) Males Females 1 (Control) 0a 20.0a 2 2 2 (Low dose) 0.02 20.0b 3 3 3 (High dose) 2.0 20.0C 1 1 a The control group (Group 1) received an equivalent amount o f lactose in gelatin capsules as the total material administered to Group 3. b The low-dose group received the test material triturated with lactose (1:999, w:w). c The high-dose group received the test material triturated with lactose (1:9, w:w). Dose Preparation Gelatin capsules (Torpac, Inc., Fairfield, New Jersey), Size No. 2, Lot No. 122630 (expiration date June 26, 2002), were used for dose administration. The test material triturated with lactose and lactose only (control group) were dispensed into capsules twice weekly and stored at room temperature. 15 000151 Covance 6329-222 3M T-6295.6 Vehicle. Lactose was used as the vehicle in the dose preparations. Dose levels were based on the vehicle as supplied for Group 1. For Group 1 dose preparations, the specified amount of lactose was weighed and transferred into the gelatin capsule. The top and bottom halves of each capsule were joined, and the capsule was placed into the appropriate container labeled with study number, group number, animal number, and dose level. Test Material. For Group 2 and 3 dose preparations, the test material was dissolved in acetone and triturated with lactose (1:999 and 1:9, w:w, respectively) once before initiation o f treatment. Acetone (Spectrum, Gardena, California), Lot No. LH0253 (expiration date June 2000) was used to dilute the test material with lactose. Trituration with lactose was necessary to facilitate capsule preparation. For Group 3 dose preparations, the specified amount of test material was weighed, placed into a labeled mixing container, and the appropriate volume of acetone was added to the container. The preparation was stirred manually until the test material was dissolved. The specified amount o f lactose was weighed, placed into the mixing container, and thoroughly mixed using a spatula. This preparation was exposed to air until the acetone was evaporated and stirred periodically. Samples of the finished preparation for dose analyses were taken directly from the container. The triturated test material was transferred to a container and stored at room temperature and protected from light and moisture. For Group 2 preparations, the required amount of the Group 3 preparation was measured and placed into a labeled mixing container. The specified amount of lactose was weighed, placed into the mixing container, and thoroughly mixed using a spatula for approximately 5 minutes. Samples of the finished preparation for dose analyses were taken directly from the container. The triturated test material was transferred to a container and stored at room temperature and protected from light and moisture. The specified amount o f triturated test material was weighed and transferred into the gelatin capsule. The top and bottom halves of each capsule were joined, and the capsule was placed into the appropriate container labeled with study number, group number, animal number, and dose level. 000152 16 Covance 6329-222 3M T-6295.6 Dose Analyses Samples (1 g each) were collected from the top, middle, and bottom o f the test material/lactose preparations for the low- and high-dose groups and stored at room temperature. The samples were shipped under ambient conditions to the Sponsor for analysis (see Protocol Deviations for exceptions). Homogeneity samples collected from the middle o f the preparations were used for the prestudy stability analysis. One set o f samples (1 g each) were taken from the low- and high-dose test material/lactose preparations at the end of the in-life phase and shipped to the Sponsor for analysis (see Protocol Deviations for exceptions). Results o f the homogeneity, stability, and dose confirmation analyses provided by the Sponsor are in Appendix 7. Method of Administration Gelatin capsules were used for test material preparation administration to compare with data from previous toxicology studies using the oral route, which is the most likely route of exposure. The dose preparations were administered orally in gelatin capsules once daily (7 days/week) for 29 days (see Protocol Deviations for exceptions). The dosing tubes were flushed with 5 mL of reverse osmosis water. Individual doses were calculated based on the most recently recorded body weights, with the exception o f body weight collection days when the previous body weight was used. Animals were dosed at approximately the same time each day. Clinical Observations The animals were observed twice daily (a.m. and p.m.) for mortality and moribundity. Each animal was observed daily, and food consumption was assessed qualitatively; abnormal findings were recorded. Animals were observed approximately 30, 60, and 90 minutes postdose for signs o f poor health or abnormal behavior. Effects were recorded as they were observed. Animals were observed once weekly; abnormal findings or an indication o f normal was recorded. 000153 17 Covance 6329-222 3M T-6295.6 Body Weights Individual body weight data were recorded twice weekly beginning 2 weeks before initiation o f treatment (Monday and Thursday), on the first day of treatment, and twice weekly thereafter. An additional body weight was recorded on Day -1 for the Day 1 dose calculations. Rectal Body Temperatures Rectal body temperatures were taken at approximately 11:00 twice weekly beginning 2 weeks before initiation o f treatment [Monday and Thursday (see Protocol Deviations for exceptions)]. Animals were not anesthetized for the procedure. Blood Hormone Determination Approximately 5 mL of whole blood were collected from a femoral vein of each animal before initiation o f treatment (Day -7) and predose on Day 29. Blood was collected between 07:00 and 09:00. Animals were fasted overnight before collections. All samples were collected without anticoagulant, maintained at room temperature, and allowed to clot. Samples were centrifuged within 1 hour after collection, and serum samples were harvested. Serum was divided into two approximately equal aliquots (blood collected on Day 29 only) and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to Ani Lytics Inc., for analyses. The samples were analyzed for estradiol, estrone, estriol, thyroid stimulating hormone, triiodothyronine (T3), and thyroxin (T4). Results o f these analyses were provided by Ani Lytics Inc., and are in Appendix 6. Serum PFOS Level Determination Approximately 5 (Day -7 only) or 2 mL o f whole blood were collected from a femoral vein o f each animal before initiation o f treatment (Day -7) and predose on Days 2, 3 (approximately 24 hours after the first and second dose, respectively), 7, 14, and 29. Animals were fasted overnight before collections. All samples were collected without anticoagulant, maintained at room temperature, and allowed to clot. Samples were centrifuged within 1 hour after collection, and serum was harvested and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to the Sponsor for analyses. The samples were analyzed for PFOS. Results o f these analyses will be reported separately by the Sponsor. 000154 18 Covance 6329-222 3M T-6295.6 Clinical Pathology Blood samples were collected from each animal on Days -7 and 29 for hematology and clinical chemistry tests. In addition, blood for clinical chemistry tests was collected from each animal predose on Days 2, 7, and 14. Animals were fasted overnight; water was provided ad libitum. Blood was collected from the femoral vein. The following were evaluated: Hematology red blood cell (erythrocyte) count differential blood cell count hemoglobin hematocrit segmented neutrophil count lymphocyte count mean corpuscular volume monocyte count mean corpuscular hemoglobin eosinophil count mean corpuscular hemoglobin concentration basophil count platelet count white blood cell (leukocyte) count blood cell morphology reticulocyte count glucose urea nitrogen creatinine total protein albumin globulin total bilirubin cholesterol triglycerides aspartate aminotransferase alanine aminotransferase alkaline phosphatase Clinical Chemistry gamma glutamyl transferase sorbitol dehydrogenase creatine kinase calcium inorganic phosphorus sodium potassium chloride serum bile acids amylase lipase pancreatic-specific amylase Additional Blood Collection Whole blood (approximately 20 mL) was collected from the vena cava of each animal at the time o f exsanguination (See Protocol Deviations for exceptions). The anticoagulant was potassium EDTA. Samples were transferred into containers (approximately 5 mL each) and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to the Sponsor possible future analysis. 000155 19 Covance 6329-222 3M T-6295.6 Necropsy On Day 30, animals that were fasted overnight were anesthetized with ketamine and xylazine, weighed, bled for required tests, exsanguinated, and necropsied. Animals were necropsied in random order. The necropsy included a macroscopic examination o f the external surface o f the body; all orifices; the cranial cavity; the brain and spinal cord; the nasal cavity and paranasal sinuses; cervical tissues and organs; and the thoracic, abdominal, and pelvic cavities and viscera. Palmitoyl CoA Oxidase Determinations A sample o f the right lateral lobe o f liver was collected from each animal at scheduled sacrifice. The sample was flash-frozen in liquid nitrogen and stored in a freezer set to maintain -60 to -80C until analyzed for palmitoyl CoA oxidase activity. Cell Proliferation Evaluation Representative samples o f the liver, testes, and pancreas were collected and preserved in zinc formalin. After fixation, samples for proliferation cell nuclear antigen (PCNA) evaluation were embedded in paraffin and shipped to Pathology Associates International for PCNA evaluation. PCNA evaluation (including examination of slides stained with hematoxylin and eosin) was done on the samples. Results of the evaluation provided by Pathology Associates International are in Appendix 8. Liver PFOS Determination A sample o f liver was collected from each animal at sacrifice, weighed, flash-frozen in liquid nitrogen, and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped with plasma samples to the Sponsor for PFOS analysis. Results of these analyses will be reported separately by the Sponsor. 000156 20 Covance 6329-222 3M T-6295.6 Tissue Preservation The following tissues (when present) or representative samples were collected and preserved in 10% neutral-buffered formalin, unless otherwise specified, for possible future microscopic examination: adrenal (2) aorta brain cecum cervix colon duodenum epididymis (2) esophagus eyes [preserved in Davidson's fixative (2)] femur with bone marrow (articular surface o f the distal end) gallbladder heart ileum jejunum kidney (2) liver lung lymph node (mesenteric) mammary gland muscle (thigh) ovary (2) pancreas pituitary prostate rectum salivary gland [mandibular (2)] sciatic nerve seminal vesicle (2) skin spinal cord (cervical, thoracic, and lumbar) spleen sternum with bone marrow stomach testis (2) thymus thyroid (2) with parathyroid trachea urinary bladder uterus vagina Histopathology The adrenals, eye, femoral bone marrow, lung, liver, kidney, pancreas, spleen, testes, and thymus were embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically from each animal (see Protocol Deviations for exceptions). Bone marrow smears from the sternum of each animal at the scheduled sacrifice were prepared, stained with Wright's stain, and retained for possible examination. 000157 21 Covance 6329-222 3M T-6295.6 Statistical Analyses Only data collected on or after the first day of treatment were analyzed statistically. Statistical analyses were not done on data collected for Group 3 due to the small number o f animals. One-way analysis o f variance [ANOVA (Winer, 1971a)] was used to analyze initial body weights, rectal body temperature, palmitoyl CoA oxidase activities, and continuous clinical pathology values. Levene's test (Levene, 1960) was done to test for variance homogeneity. In the case of heterogeneity of variance at p < 0.05, transformations were used to stabilize the variance. ANOVA was done on the homogeneous or transformed data. If the ANOVA was significant, Dunnett's t-test (Dunnett, 1964) was used for control versus treated group comparisons. One-way analysis o f covariance [ANCOVA (Winer, 1971b)] was used to analyze body weights, with initial body weights as the covariate. Although Levene's test for variance homogeneity was done (see above), no transformations were used because covariance adjustment removed extraneous heterogeneity. If the ANCOVA was significant, least squares means t-test was used for control versus treated group comparisons. For each sex, Group 2 was compared with Group 1 (Control). Group comparisons were evaluated at the 5.0% two-tailed probability level. RECORD RETENTION All raw data, documentation, records, protocol, and specimens generated as a result of this study will be archived in the storage facilities o f Covance-Madison for a period of 1 year. One year after the submission o f the final report, the Sponsor will determine the final disposition o f the materials. All raw data stored on magnetic media and an original copy o f the final report will be retained by Covance-Madison. 22 0001S8 Covance 6329-222 3M T-6295.6 Within 1 year after submission o f the final report, all o f the aforementioned materials (as appropriate) from the Sponsor's designees (3M E.T. & S and Ani Lytics Inc.) will be sent to the Sponsor (Andrew Seacat, PhD, 3M). Pathology Associates International (PAI) is responsible for the maintenance of any raw data or specimens produced by PAL RESULTS Dose Analyses Results of analyses provided by the Sponsor are in Appendix 7. Dose analysis results indicate that the average + standard deviation for the 2.0 mg/kg/day dose level prepared at a 1:9 dilution was 80 + 02% o f target. The average + standard deviation for the 0.02 mg/kg/day dose level prepared at a 1:999 dilution was 54% + 02% of target. The stability samples obtained from the middle of each mixture on May 22, 1998, were 79% and 71% of target for the 2.0 and 0.02 mg/kg/day preparations, respectively. Inherent variability and lack o f sufficient sensitivity in the analytical method for measuring (PFOS; T-6295) resulted in stability and routine analysis data that were outside of the generally recognized as acceptable standard limits o f + 15%. Clinical Observations Clinical observations are summarized in Tables 1 through 5; individual data are in Appendix 2. All animals survived to the scheduled sacrifice. There were no test material-related clinical observations. Body Weights Body weight data are summarized in Table 6; individual data are in Appendix 3. There were no apparent test material-related effects on body weights. 23 0003,59 Covance 6329-222 3M T-6295.6 Food Consumption Food consumption data are summarized in Table 1 (Summary of Clinical Observations - A.M./Weekly); individual data are included in the individual clinical observations in Appendix 2. There were no test material-related effects on food consumption. Rectal Body Temperatures Rectal body temperature data are summarized in Tables 7; individual data are in Appendix 2. There were no test material-related effects on rectal body temperatures. Blood Hormone Analyses Summary and individual hormone analyses data are in Appendix 6. Estradiol and triiodothyronine analyses data are illustrated in Figures 1 through 4 (Appendix 6). There were no apparent test material-related effects on estrone, estriol, thyroid stimulating hormone, or thyroxin. There were no apparent test material-related effects on estradiol or triiodothyronine at 0.02 mg/kg/day. Given the low number o f animals in this study, it is not possible to determine conclusively whether there was a test material-related effect on estradiol or triiodothyronine. Estradiol levels for the male and female given 2.0 mg/kg/day were numerically lower than those of the control animals on Day 29 and their respective prestudy values. Triiodothyronine levels for the male and female given 2.0 mg/kg/day were numerically lower than those of the control animals on Day 29; however, their prestudy values were also considerably lower than the control values. Triiodothyronine levels for animals given 0 or 0.02 mg/kg/day were numerically higher on Day 29 compared with their prestudy levels while triiodothyronine levels for animals given 2.0 mg/kg/day were numerically lower compared with their prestudy levels. 000160 24 Covance 6329-222 3M T-6295.6 Clinical Pathology Hematology and clinical chemistry data are summarized in Tables 8 through 15; individual data are in Appendix 4. The Pathology Report contains a discussion of the data. Administration of PFOS had no effects on clinical pathology test results. Cell Proliferation Evaluation Results o f cell proliferation evaluation provided by Pathology Associates International are in Appendix 8. Cell proliferation, as determined by measuring the proliferating index, was not increased in the liver, testes, or pancreas o f monkeys. Anatomical Pathology Incidences o f macroscopic and microscopic observations are summarized in Tables 16 and 17. Individual data are in Appendix 5. The Pathology Report contains a discussion o f the data. There were no test material-related macroscopic or microscopic changes in the organs and tissues examined. CONCLUSIONS In cynomolgus monkeys treated with 0, 0.02, or 2.0 mg PFOS/kg/day orally for at least 4 weeks, there were no apparent test material-related effects on clinical observations, body weights, food consumption, rectal body temperatures, or clinical or anatomical pathology findings. The one male and one female treated with 2.0 mg PFOS/kg/day appeared to have lower levels o f estradiol on Day 29. 000161 25 SIGNATURES Patricia K. McKee Pesik, BS, LAT Study Coordinator Covance Laboratories Inc. iter J. Thomfpra, PhD Study Direefer Covance Laboratories Inc. Date Covance 6329-222 3M T-6295.6 000162 Covance 6329-222 _______________________________________________________________________________3M T-6295.6 REFERENCES Dunnett, C. W., "New Tables for Multiple Comparisons with a Control," Biometrics. 20:482-491 (1964). Levene, H., "Robust Tests for Equality o f Variances," Contributions to Probability and Statistics, (eds.) I. Olkin et al., Ch. 25, pp. 278-292, Stanford University Press: Stanford, California (1960). Winer, B. J., "Design and Analysis o f Single-Factor Experiments," Statistical Principles in Experimental Design. Second Ed., Ch. 3, pp. 149-260, McGraw-Hill: New York, New York (1971a). Winer, B. J., "Analysis of Covariance," Statistical Principles in Experimental Design. Second Ed., Ch. 10, pp. 752-812, McGraw-Hill: New York, New York (1971b). 000163 27 PATHOLOGY REPORT Covance 6329-222 3M T-6295.6 SUMMARY The purpose o f this study was to provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study and to assess the effect of the test material, Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295), on critical enzyme levels, hormones, and other selected biochemical parameters. Administration o f PFOS had no effects on clinical pathology test results and caused no test material-related macroscopic or microscopic changes in the organs and tissues examined. METHODS Three groups o f male and female cynomolgus monkeys were administered the test material orally via capsule at a dose level of 0 (control group; received the vehicle, lactose), 0.02, or 2.0 mg/kg o f body weight/day (mg/kg/day) for 29 days. The control group had two animals/sex, the low-dose group had three animals/sex, and the high-dose group had one animal/sex. Blood was collected for hematology and clinical chemistry tests before initiation of treatment (Day -7) and on Day 29. In addition, blood for clinical chemistry tests was collected on Days 2, 7, and 14. The animals were sacrificed and necropsied on Day 30; macroscopic observations were recorded, and tissues were preserved in fixative as specified by the protocol. Samples of liver were collected and frozen for determination of hepatic palmitoyl CoA oxidase activity and for determination o f PFOS content (by the Sponsor). Samples o f liver, testes, and pancreas were collected and preserved in zinc formalin for evaluation o f proliferation cell nuclear antigen (PCNA; by Pathology Associates International). Microscopic examinations were done on adrenals, eye, femoral bone marrow, lung, liver, kidney, pancreas, spleen, testes, and thymus from all animals. Results o f clinical pathology tests for the control and low-dose groups were compared statistically. 000164 28 RESULTS AND DISCUSSION Mortality All animals survived to the scheduled sacrifice. Covance 6329-222 3M T-6295.6 Clinical Pathology Individual values for hematology and clinical chemistry tests are in Appendix 4. Mean values and the results of statistical comparisons are in Tables 8 through 15. Day -7. Clinical pathology test results indicated no obvious group or individual health abnormalities. Days 2, 7, and 14. Administration o f the test material had no effects on clinical chemistry test results at these collection intervals. Statistically significant differences for clinical chemistry results between the control and low-dose groups were inconsistent over time and between the sexes. Several of the differences were similar to differences present at Day -7, and none o f the differences exhibited a strong dose relationship (i.e., the high-dose animals did not exhibit notable differences for the same parameters). Day 29. Administration o f the test material had no effects on hematology or clinical chemistry test results at Day 29. Statistically significant differences for red blood cell count, neutrophil count, monocyte count, and hepatic palmitoyl CoA oxidase were inconsistent between the sexes and exhibited no dose relationship. Statistically significant differences for triglycerides and alkaline phosphatase were similar to differences observed at Day -7. Anatomical Pathology Anatomical pathology findings for each animal are in Appendix 5. Incidence summaries o f the macroscopic and microscopic observations are in Tables 16 and 17. U0015 29 Covance 6329-222 3M T-6295.6 Week 5. There were no macroscopic and few microscopic findings in the organs and tissues examined, and there were no major differences between control and treated animals in the incidence of any finding. All microscopic observations were considered incidental and unrelated to the test material. Robert L. Hall, DVM, PhD Diplomate, ACVP (Clinical Pathology) J-- \ \ XamV W i a Dawn~0: Goodman,\VMD Diplomate, ACVPx^X Director of Pathology.'Tlorth America a / a ia t J .M 3 - Date ( i t y W i. . ^ C O -X Date 000166 30 COMMENTS ON THE DATA Covance 6329-222 3M T-6295.6 Various models o f calculators, computers, and computer programs were used to analyze data in this study. Because different models round off or truncate numbers differently, values in some tables (e.g., means, standard deviations, or individual values) may differ slightly from those in other tables, from individually calculated data, or from statistical analysis data. Neither the integrity nor the interpretation of the data was affected by these differences. Some tabular data were compiled using Excel Version 7.0 software. The summary tables for clinical observations indicate the number of animals for which a condition was observed without regard to the specific nature, severity, reversibility, number of incidences/animal, or the length of time the condition persisted. Only observations other than normal are indicated on the summary clinical observations tables. The day o f initiation o f treatment is "Day 1." 000167 31 CODES, ABBREVIATIONS, AND UNITS Covance 6329-222 3M T-6295.6 General Codes and Abbreviations Codes for Clinical Pathology Abbreviations and Units for Clinical Hematology Abbreviations and Units for Clinical Chemistry Codes for Anatomical Pathology Note: The following lists of codes, abbreviations, and units are used by Covance. Some, but not necessarily all, o f this information may be needed for this report. 000168 32 Covance 6329-222 3MT-6295.6 General Codes and Abbreviations WK N Mean; MEAN CAM SD; S.D.; STAND DEV; STANDARD DEV; sd * -; NA P UNSCHED DISPATCH MIN OBS Week. Number o f measurements in a group. Arithmetic mean. Covariate-adjusted mean. Standard deviation. Group 2 mean is significantly different from the mean o f the control group (Group 1) at p < 0.05. No value; not applicable; not present. Present. Unscheduled. Observations transferred from the in-life module o f the data collection system to the necropsy module for reference during necropsy. Observations are duplicates of the last in-life observations. Minute. Observation. Animal Death Codes: T Terminal sacrifice. 000169 33 NS QS/QNS NR FS SC SH H SL L SI I NF U DT/DOT DB TJ TE RE EE SE PC PD PI PL PA CO HB PLASMO NOAGG FR UTD NO COAG Covance 6329-222 3M T-6295.6 Codes for Clinical Pathology GENERAL CODES No sample Quantity not sufficient No repeat (sample volume not sufficient for repeat analysis) Fibrin strands Sample clotted Slightly hemolyzed Hemolyzed Slightly lipemic Lipemic Slightly icteric Icteric Animal not fasted Unscheduled/moribund bleed Animal died on test Died during bleeding Technician judgment to repeat test Technical error (instrument or technician error that results in unacceptable data, e.g., unacceptable instrument output, sample spilled, entry of invalid data) Recording error (recorded incorrect data, e.g., wrong number, spelling error, incorrect date) Entry error (incorrect keyboard entry) Sampling error Platelets clumped Platelets decreased Platelets increased Platelets large Platelets appear adequate Color interferes with test Heinz bodies observed Plasmodium No aggregation Fractious Unable to determine No coagulation 34 000170 Codes for Clinical Pathology (Continued) Covance 6329-222 3M T-6295.6 RESULTS NOT INCLUDED IN STATISTICAL ANALYSES Hemolyzed clinical chemistry or coagulation samples Samples from animals at unscheduled intervals Prothrombin times (PT) greater than 50 seconds Activated partial thromboplastin times (PTT) greater than 110 seconds Bleed times (BLETIME) greater than 30 minutes CODES FOR BLOOD CELL MORPHOLOGY The following scale was used to measure the degree o f anisocytosis (ANISO), poikilocytosis (POIK), polychromasia (POLY), hypochromasia (HYPO), and toxic neutrophils (TOXNEUT): Scale - 1 2 3 4 Degree Normal for the species Slight Moderate Marked Not applicable Presence Not present Rare Few Moderate Many 000171 35 Covance 6329-222 3M T-6295.6 Abbreviations and Units for Clinical Hematology Test Red blood cell count Hemoglobin Hematocrit Mean corpuscular volume Mean corpuscular hemoglobin Mean corpuscular hemoglobin concentration Platelet count Mean platelet volume Reticulocyte count Absolute reticulocyte count Heinz body count Erythrocyte sedimentation rate Prothrombin time Activated partial thromboplastin time Thrombin time Activated coagulation time Fibrinogen Fibrin/fibrinogen degradation products Platelet aggregation Collagen Adenosine diphosphate Alpha 2-antiplasmin Bleeding time Methemoglobin Plasma hemoglobin Myeloid/erythroid ratio Estimated myeloid/erythroid ratio White blood cell count Differential blood cell count Nucleated red blood cell count Corrected white blood cell count Segmented neutrophil count Band neutrophil count Lymphocyte count Monocyte count Eosinophil count Basophil count Anisocytosis Polychromasia Abbreviation (Units) RBC (E6/UL or X106/pL) HGB (G/DL) HCT (%) MCV (FL) MCH (PG) MCHC (%) PLT (E3/UL or X103/pL) MPV (FL) RETIC (%) RETIC (E3/UL or X103/pL) HEINZ (%) ESR (MM/HR) PT (SEC) PTT (SEC) TT (SEC) ACT (SEC) FBR (MG/DL) FDP (UG/ML) PAGG/COL (%) PAGG/ADP (%) ANTIPLAS (%) BLE TIME (SEC) METHGB (%) PLA HGB (MG/DL) M/E RATIO EST M/E RATIO WBC (E3/UL or X103/pL) NRBC (/100 WBC) COR WBC (E3/UL or X103/pL) N-SEG (E3/UL or X103/pL) and % N-BAND (E3/UL or X103/pL) and % LYMPH (E3/UL or X103/pL) and % MONO (E3/UL or X103/pL) and % EOSIN (E3/UL or X103/pL) and % BASO (E3/UL or X103/pL) and % ANISO (-,1,2,3) POLY (-,1,2,3) 000172 36 Covance 6329-222 3M T-6295.6 Abbreviations and Units for Clinical Hematology (Continued) Test Poikilocytosis Hypochromasia Howell-Jolly bodies Basophilic stippling Toxic neutrophils Atypical lymphocytes Aqueous white blood cell count (right eye) Aqueous white blood cell count (left eye) Abbreviation (Units) POIK (-,1,2,3) HYPO (-,1,2,3) HJBODY (-,1,2,3,4) BASTIP (-,1,2,3) TOXNEUT (-,1,2,3,4) ATYPLYM (-,1,2,3,4) R EYE (WBC/UL) L EYE (WBC/UL) 000173 37 Covance 6329-222 3M T-6295.6 Abbreviations and Units for Clinical Chemistry Test Glucose Urea nitrogen Urea Creatinine Total protein Albumin Globulin Albumin/globulin ratio Total bilirubin Direct bilirubin Indirect bilirubin Cholesterol Triglyceride Urea nitrogen/creatinine ratio Total lipids Phospholipids High-density lipoprotein cholesterol Low-density lipoprotein cholesterol Uric acid Aspartate aminotransferase Alanine aminotransferase Alkaline phosphatase Gamma glutamyl transferase Sorbitol dehydrogenase Lactate dehydrogenase Creatine kinase Amylase Lipase Palmitoyl CoA oxidase Calcium Ionized calcium Inorganic phosphorus Sodium Potassium Chloride Magnesium Zinc Strontium Iron Abbreviation (Units) GLU (MG/DL) UN (MG/DL) UREA (MG/DL) CREAT (MG/DL) T PRO (G/DL) ALB (G/DL) GLOB (G/DL) A/G RATIO T BILI (MG/DL) D BILI (MG/DL) I BILI (MG/DL) CHOL (MG/DL) TRIG (MG/DL) UN/CREAT (RATIO) T LIPIDS (MG/DL) P LIPIDS (MG/DL) HDL (MG/DL) LDL (MG/DL) UA (MG/DL) AST/SGOT (IU/L) ALT/SGPT (IU/L) ALK PHOS (IU/L) GGT (IU/L) SDH (IU/L) LDH (IU/L) CK (IU/L) AMYLASE (IU/L) LIPASE (IU/L) PCOAO (IU/G) CA (MG/DL) ION CA (MG/DL) I PHOS (MG/DL) NA (MMOL/L) K (MMOL/L) CL (MMOL/L) MG (MEQ/L or MG/DL) ZN (MG/L or PPM) SR (MG/L or PPM) FE (UG/DL) 38 000174 Covance 6329-222 3M T-6295.6 Abbreviations and Units for Clinical Chemistry (Continued) Test Excess iron Total iron binding capacity Unbound iron binding capacity Percent iron saturation Plasma cholinesterase Red blood cell cholinesterase Brain cholinesterase Caudate putamen Hippocampus Frontal cortex Cerebellum Bicarbonate Serum hemoglobin Serum bile acids Fecal bile acids Average fecal weight Fecal bile acids (calculation) Osmolality Electrophoresis Albumin A lp h a -1-globulin Alpha-2-globulin Beta globulin Gamma globulin High-density lipoprotein Low-density lipoprotein Very-low-density lipoprotein Insulin Adrenocorticotropic hormone Cortisol Glucagon Triiodothyronine Thyroxine Pancreatic-specific amylase Creatine kinase isoenzymes BB MB MM Abbreviation (Units) EX FE (UG/DL) TIBC (UG/DL) UIBC (UG/DL) FE %SAT (%) CHEP (MU/ML) CHER (MU/ML) CHEB (MU/ML) CAUD PUT (UMOL/G) HIPPOCAM (UMOL/G) F CORTEX (UMOL/G) CEREBELL (UMOL/G) BICARB (MMOL/L) SER HGB (MG/DL) SBA (UMOL/L or MG/DL) FBA (UG/ML) FCC WGT (G) FBA (MG/Day) OSMO (MOSM/KG) E ALB (G/DL) E A -l (G/DL) E A-2 (G/DL) E BETA (G/DL) E GAMMA (G/DL) E-HDL (%) E-LDL (%) E-VLDL (%) INSULIN (UU/ML) ACTH (PG/ML) CORTISOL (UG/ML) GLUCAGON (PG/ML) T3 (NG/DL) T4 (UG/DL) P AMYL (U/L) CK-BB (U/L) CK-MB (U/L) CK-MM (U/L) 39 000 Covance 6329-222 3M T-6295.6 Codes for Anatomical Pathology MICROSCOPIC CODES Distribution of Findings Focal Diffuse Multifocal Grades for Severity or Amount 1 Minimal - the least amount o f change that can be observed with the light microscope 2 Slight - less than average amount of change, but readily discernible as abnormal 3 Moderate - the average amount o f change that is expected for a lesion 4 Moderately severe (marked) - a marked amount of change with possible loss of function of the affected cells or organs 5 Severe - a great amount o f change with probable loss o f function of the affected cell or organs and frequently involves large areas of the organ 000176 40 iTOOO Table 1 Summary of Clinical Observations A.M./Weekly 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS DAYS 1-30 CATEGORY KEYWORD QUALIFIER *** TOP OF LIST *** APPEARANCE SWOLLEN LIP(S) DISCHARGE VOMITUS CONTAINING FOOD APPEARS TO BE MENSTRUATING DISCHARGE UNKNOWN SOURCE FOUND IN PAN RED IN COLOR EXCRETION FEW FECES NON-FORMED FECES SKIN & PELAGE RED SKIN LIP(S) SCAB(S) MOUTH QUALITATIVE FOOD CONSUMPTION LOW *** END OF LIST *** SEX: GROUP: DOSE: NUMBER: NUMBER OF ANIMALS AFFECTED --MALE- -- 123 0 0.02 2 -- -FEMALE-123 0 0.02 2 231231 000 100 001000 000010 001000 000 00 1 000200 000 100 000 100 0 10 10 1 41 Covance 6329-222 3M T-6295.6 PAGE: 1 DAYS 1-29 CATEGORY KEYWORD QUALIFIER *** TOP OF LIST *** APPEARANCE SWOLLEN LIP(S) EXCRETION NON-FORMED FECES SKIN & PELAGE RED SKIN LIP(S) SCAB(S) MOUTH *** END OF LIST *** Table 2 Summary of Clinical Observations 30 Minutes Postdose 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS SEX: GROUP: DOSE: NUMBER: NUMBER OF ANIMALS AFFECTED -- MALE- -- i23 0 0.02 2 -- -FEMALE-123 0 0.02 2 2 3 12 31 000100 000 100 000100 000100 Covance 6329-222 3M T-6295.6 PAGE : 1 000178 42 DAYS 1-29 CATEGORY KEYWORD QUALIFIER *** TOP OF LIST *** APPEARANCE SWOLLEN LIP (S) EXCRETION MUCOID FECES SKIN i PELAGE RED SKIN LIP(S) SCAB(S) MOUTH *** END OF LIST *** Table 3 Summary of Clinical Observations 60 Minutes Postdose 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS SEX: GROUP: DOSE: NUMBER: NUMBER OF ANIMALS AFFECTED -- --MALE- -- 12 3 0 0.02 2 -- -FEMALE' 123 0 0.02 2 231231 0 00 100 100000 000100 000100 Covance 6329-222 3M T-6295.6 PAGE: 1 000179 43 DAYS 1-29 CATEGORY KEYWORD QUALIFIER *** TOP OF LIST *** APPEARANCE SWOLLEN LIP(S) SKIN & PELAGE RED SKIN LIP(S) SCAB(S) MOUTH *** END OF LIST *** Table 4 Summary of Clinical Observations 90 Minutes Postdose 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS SEX: GROUP: DOSE: NUMBER: NUMBER OF ANIMALS AFFECTED --MALE- -- i23 0 0.02 2 -- -FEMALE123 0 0.02 2 2 3 12 3 1 000100 000100 000100 Covance 6329-222 3M T-6295.6 PAGE: 1 QQ0180 44 DAYS 1-30 CATEGORY KEYWORD QUALIFIER *** TOP OF LIST *** DISCHARGE VOMITUS CONTAINING FOOD *** END OF LIST *** Table 5 Summary of Clinical Observations Unscheduled 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS/T-6295) IN CYNOMOLGUS MONKEYS SEX: GROUP: DOSE: NUMBER: NUMBER OF ANIMALS AFFECTED --MALE- -- 123 0 0.02 2 -- -FEMALE-12 3 0 0.02 2 2 31231 010000 Covance 6329-222 3M T-6295.6 PAGE: 1 T8TOOO 45 000X82 Table 6 Summary of Body Weight Data (kg) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS SEX: --------------- MALE--------------- -------------- FEMALE-------------- GROUP: 1 2 3 1 2 3 DOSE: 0 0.02 2 0 0.02 2 DAY UNITS: MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY -14 N MEAN S.D. -10 N MEAN S.D. -7 N MEAN S.D. -3 N MEAN S.D. -1 N MEAN S.D. 1N MEAN S.D. 5N CAM MEAN S.D. 2 2.3 0.00 2 2.3 0.07 2 2.3 0.07 2 2.3 0.07 2 2.3 0.00 2 2.3 0.14 2 2.3 2.3 0.07 3 2.3 0.21 3 2.3 0.21 3 2.3 0.21 3 2.3 0.21 3 2.4 0.25 3 2.3 0.21 3 2.3 2.3 0.17 1 2.2 1 2.2 1 2.2 1 2.2 1 2.2 1 2.2 1 2.2 2 2.1 0.07 2 2.1 0.07 2 2.1 0.00 2 2.1 0.00 2 2.1 0.00 2 2.1 0.00 2 2.0 2.0 0.00 3 2.1 0.17 3 2.1 0.10 3 2.1 0.15 3 2.1 0.15 3 2.1 0.20 3 2.2 0.15 3 2.1 2.1 0.15 1 2.2 1 2.1 1 2.2 1 2.1 1 2.2 1 2.2 1 2.1 46 Covance 6329-222 3M T-6295.6 PAGE: 1 oooi8a Table 6 Summary of Body Weight Data (kg) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS SEX: - - M A L E ----- --FEMALE--- GROUP: 1 2 3 1 2 3 DOSE: 0 0.02 2 0 0.02 2 DAY UNITS: MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY 8N CAM MEAN S.D. 12 N CAM MEAN S.D. 15 N CAM MEAN S.D. 19 N CAM MEAN S.D. 22 N CAM MEAN S.D. 26 N CAM MEAN S.D. 29 N CAM MEAN S.D. 2 2.5 2.5 0.07 2 2.5 2.5 0.07 2 2.4 2.4 0.14 2 2.5 2.5 0.07 2 2.5 2.5 0.07 2 2.5 2.5 0.07 2 2.4 2.4 0.07 3 2.4 2.4 0.12 3 2.3* 2.3 0.12 3 2.4 2.4 0.06 3 2.4 2.4 0.06 3 2.4 2.4 0.10 3 2.4 2.4 0.10 3 2.4 2.4 0.06 1 2.3 1 2.2 1 2.3 1 2.3 1 2.1 1 2.1 1 2.0 2 2.2 2.2 0.07 2 2.1 2.1 0.00 2 2.2 2.2 0.07 2 2.1 2.1 0.00 2 2.2 2.2 0.07 2 2.2 2.2 0.07 2 2.1 2.1 0.00 3 2.2 2.2 0.20 3 2.1 2.2 0.15 3 2.2 2.2 0.20 3 2.3 2.3 0.17 3 2.2 2.2 0.15 3 2.2 2.2 0.15 3 2.1 2.1 0.15 1 2.2 1 2.2 1 2.1 1 2.3 1 2.1 1 2.1 1 2.1 47 Covance 6329-222 3M T-6295.6 PAGE: 2 000184 Table 7 Summary of Rectal Body Temperature Data (C) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS SEX: --------------- MALE--------------- -------------- FEMALE GROUP : i 2 3 1 2 3 DOSE : 0 0.02 2 0 0.02 2 DAY UNITS: MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY -14 N MEAN S.D. -10 N MEAN S.D. -7 N MEAN S.D. -3 N MEAN S.D. 1N MEAN S.D. 5N MEAN S.D. 8N MEAN S.D. 2 39.8 0.28 2 40.0 0.35 2 39.8 0.57 2 39.6 0.28 2 39.0 0.57 2 38.9 0.07 2 38.5 0.35 3 39.2 0.57 3 39.2 0.40 3 39.0 0.72 3 39.2 0.45 3 39.4 0.30 3 39.1 0.59 3 39.2 0.40 1 39.6 1 39.5 1 39.1 1 39.2 1 39.4 1 39.9 1 39.6 2 40.2 0.28 2 39.8 0.49 2 39.9 0.21 2 39.8 0.49 2 39.6 0.35 2 39.0 0.35 2 39.1 0.42 3 40.1 0.50 3 39.7 0.55 3 40.0 0.38 3 39.8 0.10 3 39.9 0.29 3 39.8 0.25 3 39.8 0.21 1 40.0 1 39.8 1 39.9 1 39.7 1 39.5 1 39.7 1 39.9 Covance 6329-222 3M T-6295.6 PAGE: 1 00018S 49 o o 000186 Table 7 Summary of Rectal Body Temperature Data (C) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS SEX: --------------- MALE--------------- -------------- FEMALE GROUP : 1 2 3 1 2 3 DOSE: 0 0.02 2 0 0.02 2 DAY UNITS: MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY MG/KG/DAY 12 N MEAN S.D. 15 N MEAN S.D. 19 N MEAN S.D. 22 N MEAN S.D. 26 N MEAN S.D. 29 N MEAN S.D. 2 39.0 0.07 2 39.7 0.00 2 38.7 0.21 2 39.4 0.21 2 38.8 0.07 2 38.5 0.42 3 39.1 0.47 3 39.7 0.42 3 38.7 0.64 3 39.5 0.57 3 39.2 0.38 3 38.8 0.35 1 1 39.7 1 38.9 1 39.4 1 39.2 1 38.9 2 39.2 0.14 2 39.7 0.57 2 39.0 0.35 2 39.6 0.42 2 39.0 0.35 2 38.8 0.14 3 39.8 0.25 3 40.0 0.20 3 39.3 0.21 3 39.8 0.25 3 39.6 0.15 3 39.1 0.38 1 39.7 1 39.8 1 39.3 1 39.4 1 39.1 1 39.2 Covance 6329-222 3M T-629S.6 PAGE: 2 50 Table 8 Summary of Clinical Hematology Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE RBC HGB HCT MCV MCH MCHC PLT RETIC RETIC mg/kg/day XlOVjiL G/DL FL PG % x i o V ml % XlOVnL 0 MEAN S.D. N 0.02 MEAN S.D. N 5.64 .332 2 5.18 .272 3 14.0 1.41 2 12.6 .31 3 42.6 3.82 2 39.1 .90 3 75.4 2.26 2 75.4 2.21 3 24.8 1.06 2 24.4 1.10 3 32.9 .42 2 32.4 .91 3 502 138.6 2 454 88.1 3 .9 .00 2 .9 .53 3 51 2.8 2 46 26.1 3 MEAN 5.91 12.8 41.0 69.4 21.7 31.2 531 .7 41 N 1111111i1 ro o 000187 51 Table 8 Summary of Clinical Hematology Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE mg/kg/day WBC XIOVuL N-SEG XlOVnL LYMPH XIOVuL MONO XIOVuL EOS IN xioV ml BASO XIOVuL N-SEG% LYMPHi MONO% EOSIN% BASO% 0 MEAN 8.4 3.4 4.6 .4 .1 .0 40 54 4 10 S.D. .99 1.06 .07 .07 .14 .00 7.8 7.1 .7 1.4 .0 N 22222222222 0.02 MEAN 9.2 2.9 5.8 .4 .1 .0 31 63 4 1 0 S.D. 1.00 1.33 1.46 .20 .10 .00 16.5 14.6 2.0 1.0 .0 N 33333333333 2.0 MEAN 8.2 2.8 4.8 .6 .0 .0 34 58 800 N 11111111111 88T000 52 Table 8 Summary of Clinical Hematology Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE RBC HGB HCT MCV MCH MCHC PLT RETIC RETIC mg/kg/day X I O V h L G/DL % FL PG % X103/)lL % x i o '/hl 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 5.25 .325 2 5.00 .241 3 5.53 1 13.3 1.34 2 12.8 .21 3 13.4 1 40.4 3.96 2 39.1 .21 3 40.2 1 76.9 2.69 2 78.2 3.30 3 72.7 1 25.4 .92 2 25.6 1.55 3 24.2 1 33.1 .00 2 32.7 .56 3 33.3 1 478 180.3 2 430 121.3 3 505 1 .8 .21 2 .7 .17 3 .8 1 45 14 .1 2 35 8.5 3 44 1 Q 001& 9 53 Table 8 Summary of Clinical Hematology Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE WBC mg/kg/day XlOVjiL 0 MEAN S .D . N 9.0 1.48 2 0.02 MEAN S.D. N 10.2 1.97 3 2.0 MEAN N 11.7 1 N-SEG XlO'/pL LYMPH X10V|iL 3.6 1.91 2 5.1 .14 2 3.5 3.29 3 6.2 4.05 3 6.2 4.8 11 MONO XlOVpL EOSIN XIOV^L BASO xioV hl N-SEG% LYMPH% MONO% EOSIN% BASO% .2 .0 .0 38 58 2 0 0 .28 .07 .00 14.8 11.3 3.5 .7 .0 22 222222 .4 .1 .0 36 59 4 1 0 .12 .10 .00 34.2 32.9 1.5 1.0 .0 33 333333 .4 .2 .0 53 42 4 1 0 1 1 11 1111 OGTOQO 54 Table 9 Summary of Clinical Hematology Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE RBC HGB HCT MCV MCH MCHC PLT RETIC RETIC mg/kg/day X10 VflL G/DL % FL PG % XlOVjiL % x i o V hl 0.02 MEAN S. D. N MEAN S.D. N MEAN N 6.02 .035 2 14.0 .64 2 5.12 * .262 3 12.6 .78 3 5.69 1 13.2 1 44.0 1.84 2 39.1 2.18 3 41.5 1 72.9 2.69 2 76.4 1.69 3 73.1 1 23.3 .99 2 24.6 .61 3 23.1 1 31.9 .21 2 32.2 .26 3 31.7 1 440 116.7 2 392 92.1 3 503 1 .8 .14 2 .5 .35 3 .2 1 48 8.5 2 28 19.8 3 11 1 T6T000 55 Table 9 Summary of Clinical Hematology Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE mg/kg/day WBC XIOVuL N-SEG x i o V nl LYMPH xioV nl MONO XIOVuL EOS IN XIOVuL BASO X103/r L N-SEG% LYMPH* MONO* EOSIN* BASO* 0 MEAN 9.2 1.6 6.8 .6 .2 .0 18 74 6 2 0 S.D. .21 .49 .78 .21 .14 .00 5.7 7 .1 2.8 1.4 N 22222222222 0.02 MEAN 8.8 1.0 6.9 .6 .2 .0 13 78 6 2 0 S.D. 2.44 .15 2.27 .38 .10 .00 6.1 5.5 2.3 1.0 N 33333333333 2.0 MEAN 5.8 .9 4.4 .4 .0 .0 16 77 7 0 0 N 11111111111 000192 56 Table 9 Summary of Clinical Hematology Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE RBC HGB HCT MCV MCH MCHC PLT RETIC RETIC mg/kg/day X l o V n L G/DL % FL PG x i o V hl % X10a/|iL 0.02 MEAN S.D. N MEAN S.D. N MEAN N 5.14 .170 2 5.13 .116 3 4.81 1 13.2 .78 2 13.1 .87 3 12.3 1 40.5 2.33 2 41.1 1.74 3 36.7 1 78.9 2.12 2 80.1 3.93 3 76.4 1 25.6 .71 2 25.5 1.80 3 25.6 1 32.5 .00 2 31.9 .81 3 33.6 1 511 182.4 2 425 116.7 3 529 1 1.2 .21 2 .8 .26 3 1.0 1 59 12.7 2 41 14.0 3 48 1 000193 57 Table 9 Summary of Clinical Hematology Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE mg/kg/day WBC XlOVfiL N-SEG LYMPH XlOVjiL X103/RL MONO EOS IN BASO Xl03/|iL X103/r L XlO3/JiL N-SEG% LYMPH% MONO% EOSIN% BASO% 0 00 ro MEAN li.i 6.2 1.0 .0 .0 36 54 10 0 0 S.D. 2.69 .14 2.55 .21 .00 .00 10.6 10.6 .7 .0 N 22222222222 0.02 MEAN S.D. N 10.0 2.55 3 2.2 * .45 3 7.1 2.67 3 .6 * .06 3 .1 .00 3 .0 22 70 .00 7.6 9.2 333 6 10 2 .1 .0 333 2.0 MEAN 15.7 7.2 7.9 .5 .1 .0 46 50 3 1 0 N 11111111111 ftG T O O 58 DOSE mg/kg/day GLU MG/DL Table 10 Summary of Clinical Chemistry Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN S. D. N 0.02 MEAN S. D. N 2.0 MEAN N 82 19.1 2 61 13.2 3 54 1 24 1.4 2 29 2.0 3 28 1 1.0 .07 2 1.0 .06 3 1.0 1 8.2 .00 2 7.7 .06 3 8.2 1 5.2 .00 2 4.8 .29 3 5.0 1 3.0 .00 2 3.0 .32 3 3.2 1 .2 8 155 64 .07 .7 62.2 24.7 222 2 .3 9 161 85 .06 1. 5 35.4 22.2 3333 .4 10 150 68 i111 S6T000 59 Table 10 Summary of Clinical Chemistry Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S. D. N 0.02 MEAN S.D. N 2.0 MEAN N 48 4.9 2 50 7.6 3 54 1 40 9.2 2 612 36.1 2 58 13.0 3 681 81.1 3 50 1086 11 88 4.9 2 101 36.7 3 117 1 4 228 424 .7 102.5 99.7 222 6 214 315 1.2 17.2 63.1 333 5 220 244 1 11 64 37.5 2 62 61.2 3 7 1 291 69 2 223 50 3 144 1 96T000 60 Table 10 Summary of Clinical Chemistry Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L 0 MEAN S. D. N 0.02 MEAN S.D. N 2.0 MEAN N 10.7 .14 2 10.6 .52 3 11.4 1 7.2 .57 2 7.7 .71 3 8.7 1 162 2.1 2 159 5.0 3 162 1 5.8 .07 2 5.6 .47 3 5.9 1 110 .0 2 112 4.6 3 111 1 Covance 6329-222 3M T-6295.6 000197 61 DOSE mg/kg/day GLU MG/DL Table 10 Summary of Clinical Chemistry Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN S. D. N 76 8.5 2 24 .0 2 1.0 .14 2 8.0 .49 2 5.1 .42 2 2.8 .07 2 .5 .28 2 4 142 5.7 15.6 22 76 14.1 2 0.02 MEAN S. D. N 64 26 .9 7.9 4.9 3.0 9.0 4.6 .06 .30 .26 .10 3 33 3 33 .4 .06 3 9 154 2.1 34.6 33 69 35.8 3 2.0 MEAN 71 27 1.0 8.2 4.9 3.3 .3 12 141 67 N 1111111111 86T000 62 Table 10 Sununary of Clinical Chemistry Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 99 76.4 2 58 6.7 3 53 1 53 21.2 2 58 14.2 3 55 1 593 66.5 2 828 65.5 3 683 1 80 2.1 2 74 10.1 3 110 1 6 1907 4.9 2286.8 22 4 385 1.0 163.0 33 3 1094 11 334 31.1 2 324 44.1 3 315 1 10 10.6 2 64 81.8 3 89 1 232 21.9 2 215 19.4 3 224 1 TOOO 63 Table 10 Summary of Clinical Chemistry Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 11.0 .49 2 11.0 .59 3 11.3 1 7.4 1.27 2 8.0 1.72 3 6.8 1 159 2.8 2 158 6.7 3 165 1 7.2 1.91 2 5.8 .64 3 6.0 1 113 2.8 2 114 4.0 3 115 1 Covance 6329-222 3M T-6295.6 000200 64 DOSE mg/kg/day GLU MG/DL Table 11 Summary of Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umo1/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN 57 25 .9 7.4 5.0 2.4 .4 16 142 80 S. D. 7.1 5.7 .00 .07 .07 .00 .07 4.2 64.3 48.8 N 2222222222 0.02 MEAN 62 24 .8 7.0 * 4.5 * 2.6 .3 10 * 163 69 S. D. 6.0 2.3 .06 .12 .12 .23 .06 .6 18.3 18.6 N 3333333333 2.0 MEAN 58 21 .9 7.3 4.7 2.6 .4 N 1111111 9 151 11 50 1 000201 65 Table 11 Summary of Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S. D. N 0.02 MEAN S. D. N 2.0 MEAN N 75 14.1 2 41 2.5 3 54 1 56 21.2 2 726 48.8 2 56 14.8 3 599 82.7 3 55 1028 11 84 6.4 2 88 31.8 3 105 1 2 899 412 .7 5.7 69.3 222 2 579 289 .0 391.1 51.3 333 2 219 302 111 31 5.7 2 40 19.6 3 11 1 256 40 2 180 30 3 139 1 000202 66 Table 11 Summary of Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 10.2 .14 2 9.5 * .26 3 10.0 1 6.8 1.06 2 6.7 .31 3 7.0 1 152 1. 4 2 147 2.5 3 152 1 5.2 .07 2 4.7 .25 3 5.0 1 106 .7 2 108 4.0 3 104 1 Covance 6329-222 3M T-6295.6 C02000 67 DOSE mg/kg/day GLU MG/DL Table 11 Summary of Clinical Chemistry Data Females Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN S.D. N 70 19 .8 7.4 4.9 2.5 14 .1 .0 .07 .42 .28 .14 222222 .4 .07 2 8 145 .7 2.8 22 42 11.3 2 0.02 MEAN 57 24 .8 7.3 4.7 2.6 .5 16 * 133 69 S.D. 3.5 3.5 .06 .15 .30 .23 .06 1.0 28.2 33.7 N 3333333333 2.0 MEAN 68 27 1.0 7.5 4.7 2.8 .4 17 136 47 N 1111111111 000204 68 Table 11 Summary of Clinical Chemistry Data Females Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 58 18..4 2 54 4 .5 3 46 1 72 43. 1 2 57 16. 9 3 55 1 578 123.0 2 805 65.3 3 672 1 81 5.7 2 69 8.1 3 100 1 2 523 502 .7 15.6 138.6 222 2 426 .6 133.3 33 351 46.2 3 2 308 291 111 39 5.7 2 31 9.2 3 84 1 324 96 2 208 24 3 184 1 000205 69 Table 11 Summary of Clinical Chemistry Data Females Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 10.2 .71 2 10.0 .25 3 10.0 1 6.7 .28 2 7.6 .85 3 6.5 1 153 4.2 2 151 6.2 3 151 1 5.4 .64 2 4.9 .21 3 4.7 1 108 .0 2 105 2.3 3 106 1 Covance 6329-222 3M T-6295.6 000206 70 DOSE mg/kg/day GLU MG/DL Table 12 Summary of Clinical Chemistry Data Males Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN 81 20 .8 7.8 4 .8 3.0 .2 10 138 52 S. D. 4.2 .0 .07 .28 .21 .07 .07 4.9 65.8 7 .1 N 222222222 2 0.02 MEAN 68 * 21 .8 7.5 4.5 * 3.1 .3 10 156 51 S.D. 3.1 1.5 .06 .25 .06 .25 .00 1.2 19.3 1. 5 N 3333333333 2.0 MEAN 63 18 .8 7.5 4 .6 2.9 .4 N 111111i 7 137 i1 50 1 000207 71 Table 12 Summary of Clinical Chemistry Data Males Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L 1P AMYL U/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 51 15.6 2 39 3.8 3 46 1 46 13.4 2 770 78.5 2 57 15.0 3 556 106.4 3 49 1012 11 80 .0 2 85 34.6 3 98 1 0 249 .0 79.2 22 0 399 .0 415.4 33 0 270 11 397 67.9 2 327 35.6 3 260 1 33 7.1 2 31 13.5 3 0 1 272 48.1 2 225 24.1 3 139 1 000208 72 Table 12 Summary of Clinical Chemistry Data Males Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 10.2 .35 2 9.8 .32 3 10.3 1 7.0 .49 2 7.0 .35 3 6.6 1 156 3.5 2 152 3.6 3 156 1 5.2 .21 2 4.8 .29 3 5.1 1 112 2 112 2 3 112 1 Covance 6329-222 3M T-6295.6 000209 73 DOSE mg/kg/day GLU MG/DL Table 12 Summary of Clinical Chemistry Data Females Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umo1/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0.02 MEAN S.D. N MEAN S.D. N 86 20 .8 8.0 4.6 3.5 9.2 6.4 .00 .07 .49 .42 222222 73 19 .8 8.0 4.8 3.2 4.6 1.5 .10 .17 .31 .15 333333 .2 .07 2 .3 .06 3 7 130 1.4 2.1 22 8 147 .0 31.3 33 32 13.4 2 38 6.8 3 MEAN 73 18 .8 7.9 4.6 3.3 .3 12 117 44 N 1111111111 c c c c 74 Table 12 Summary of Clinical Chemistry Data Females Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 46 .7 2 49 8.1 3 45 1 52 13.4 2 57 6.9 3 57 1 558 94.0 2 775 108.2 3 67 4 1 69 8.5 2 68 9.6 3 90 1 0 142 .7 55.2 22 0 218 .6 27.8 33 0 166 11 434 26.9 2 356 * 18.9 3 349 1 30 10.6 2 25 5.1 3 101 1 303 26.9 2 231 34.1 3 238 1 T T -0 0 0 75 Table 12 Summary of Clinical Chemistry Data Females Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 11.0 .78 2 10.7 .40 3 10.6 1 6.1 .57 2 7.3 .78 3 6.4 1 156 2.1 2 155 6.1 3 155 1 6.4 .92 2 5.5 .20 3 5.3 1 112 .7 2 113 1.5 3 113 1 Covance 6329-222 3M T-6295.6 000212 76 DOSE mg/kg/day GLU MG/DL Table 13 Summary of Clinical Chemistry Data Males Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN S. D. N 0.02 MEAN S.D. N 2.0 MEAN N 80 19 .8 7.7 5.0 2.7 13.4 1. 4 .07 .42 .28 .14 22 22 2 2 63 19 .7 7.2 4.5 2.7 3.8 1.0 .23 .45 .17 .32 333333 64 15 .7 7.4 4.8 2.6 111111 .4 .00 2 .4 .00 3 .4 1 8 141 2.1 58.0 22 8 150 2.1 12.2 33 6 132 11 38 7.1 2 44 9.5 3 27 1 000213 77 Table 13 Summary of Clinical Chemistry Data Males Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S. D. N 0.02 MEAN S. D. N MEAN N 47 7 .1 2 36 5.7 3 39 1 40 9.2 2 49 3 45 1 761 33.9 2 519 131.9 3 908 1 80 7.8 2 77 37.0 3 89 1 4 162 394 2.1 43.8 30.4 222 3 111 306 .6 35.2 40.8 333 2 245 242 111 35 8.5 2 35 18.6 3 2 1 2 62 20.5 2 203 28.9 3 123 1 o 00 o CM frTZOOO 78 Table 13 Summary of Clinical Chemistry Data Males Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L 0 MEAN S .D. N 0.02 MEAN S.D. N 2.0 MEAN N 10.2 .21 2 9.7 * .15 3 10.1 1 7.6 1.63 2 6.7 .64 3 6.4 1 151 1 .4 2 148 1.0 3 153 1 4.8 .28 2 4.5 .30 3 4.8 1 106 .7 2 107 1.0 3 108 1 Covance 6329-222 3M T-6295.6 000215 79 DOSE mg/kg/day GLU MG/DL Table 13 Summary of Clinical Chemistry Data Females Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T -6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN S. D. N 0.02 MEAN S.D. N 2.0 MEAN N 80 3.5 2 58 9.0 3 59 1 19 4.2 2 20 2.5 3 18 1 .6 7.5 .07 .14 22 .7 7.7 .20 .25 33 .8 7.8 11 4.6 .21 2 4 .8 .30 3 4.6 1 2.8 .35 2 2.9 .06 3 3.2 1 .4 .07 2 .5 .15 3 .4 1 7 119 .0 11.3 22 8 139 1.5 24.1 33 8 107 11 24 4 2 35 7 3 41 1 OOO 80 Table 13 Summary of Clinical Chemistry Data Females Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 46 8.5 2 47 7.5 3 40 1 46 7.8 2 55 11.3 3 51 1 499 58.0 2 800 * 68.6 3 727 1 64 .7 2 63 10.4 3 86 1 4 259 .7 144.2 22 428 40.3 2 30 16.3 2 288 36.1 2 3 307 320 * .6 97.5 27.2 3 33 39 24.6 3 200 * 18.8 3 3 462 440 309 283 11111 000217 81 Table 13 Summary of Clinical Chemistry Data Females Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE /day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L MEAN S. D. N 0.02 MEAN S .D. N 2.0 MEAN N 10.4 .49 2 10.4 .46 3 10.4 1 6.8 .28 2 6.7 .69 3 6.1 1 154 3.5 2 151 6.8 3 152 1 6.0 .71 2 5.2 .25 3 5.4 1 110 .7 2 108 3.6 3 108 1 Covance 6329-222 3M T-6295.6 000218 82 DOSE mg/kg/day GLU MG/DL Table 14 Summary of Clinical Chemistry Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T -6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL 0 MEAN S .D. N 81 24 .8 7.6 4 .6 3.0 8.5 .0 .07 .14 .00 .14 22 22 2 2 .2 .07 2 7 140 1. 4 65.8 22 36 6.4 2 0.02 MEAN 73 24 .8 7.5 4.3 3.2 .2 10 151 53 * S.D. 7.0 4.0 .15 1.04 .44 .65 .06 3.1 21.5 4.7 N 3333 333333 2.0 MEAN N 80 1 15 1 .1 8.2 4 .8 3.4 11111 .3 1 8 91 25 111 000219 83 Table 14 Summary of Clinical Chemistry Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L :SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S. D. N 0.02 MEAN S.D. N 2.0 MEAN N 52 12.7 2 42 7.5 3 31 1 46 14.1 2 62 26.6 3 38 1 755 60.8 2 564 127.2 3 815 1 96 9.2 2 94 42.9 3 88 1 4 410 424 2.8 93.3 87.0 222 3 314 334 1.2 255.2 57.7 333 1 202 285 111 37 .0 2 35 33.9 3 3 1 285 62.2 2 228 39.5 3 156 1 000220 84 Table 14 Summary of Clinical Chemistry Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L MEAN S .D. N 0.02 MEAN S.D. N 2.0 MEAN N 10.5 .14 2 10.2 .29 3 11.0 1 7.5 .99 2 6.3 .40 3 4 1 154 2.1 2 5.6 .92 2 111 1.4 2 152 3.0 3 5.1 .79 3 110 2.0 3 156 6.0 107 111 Covance 6329-222 3M T-6295.6 TZZOOO 85 DOSE mg/kg/day GLU MG/DL Table 14 Summary of Clinical Chemistry Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL MEAN S. D. N 86 25 .8 7.6 4 .4 3.3 .7 1.4 .07 .35 .07 .42 222222 .2 .00 2 9 121 2.8 19.8 22 38 3.5 2 0.02 MEAN S.D. N 62 24 .9 7.9 4.6 3.3 10.8 3.8 .12 .57 .38 .20 333333 .3 .10 3 9 143 1.7 24.8 33 46 6.0 3 2.0 MEAN 78 24 1.0 8.2 4.6 3.6 .2 17 62 27 N 1111111111 222000 86 Table 14 Summary of Clinical Chemistry Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 DOSE AST/SGOT ALT/SGPT ALK PHOS mg/kg/day IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN N 53 9.9 2 45 7 .1 3 36 1 46 9.2 2 54 5.7 3 47 1 467 69.3 2 908 * 68.8 3 637 1 72 3.5 2 76 7.6 3 93 1 6 298 538 130 370 3.5 51.6 133.6 157.7 99.0 22222 3 273 340 1.2 79.2 46.1 333 23 14.7 3 218 34.0 3 2 341 326 111 18 221 11 000223 87 Table 14 Summary of Clinical Chemistry Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L MEAN S.D. N 0.02 MEAN S.D. N 2. 0 MEAN N 10.4 .21 2 10.9 .46 3 11.1 1 6.4 1.06 2 6.9 .89 3 6.5 1 152 1.4 2 155 4.4 3 158 1 5.7 .42 2 5.5 .10 3 5.7 1 111 2.8 2 111 2.1 3 110 1 Covance 6329-222 3M T-6295.6 000224 88 Table 15 Summary of Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day PCOAO IU/G 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN S.D. N 0 .7 2 4* .6 3 1 .0 1 Covance 6329-222 3M T-6295.6 000225 89 Table 15 Summary of Clinical Chemistry Data Females Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS DOSE mg/kg/day PCOAO IU/G 0 MEAN S.D. N 0.02 MEAN S.D. N 2.0 MEAN S.D. N 2 2.8 2 3 3.5 3 1 .0 1 Covance 6329-222 3M T-6295.6 000226 90 TABLE 16 Incidence of Macroscopic Observations Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 1 TABLE INCLUDES: SEX=ALL;GROUP=ALL;WEEKS=ALL DEATH=T;SUBSET=ALL ORGAN AND KEYWORD(S) OR PHRASE ** TOP OF LIST ** GENERAL COMMENT (GC) .... BONE MARROW SMEAR TAKEN EYES - DAVIDSONS STAINS-PERINEUM/PERIANAL NO MACROSCOPIC LESIONS ** END OF LIST ** -- N U M B E R - O F - A N I M A L S SEX: ---- MALE---- --- FEMALE--- GROUP: -1- -2- -3- -1- -2- -3- NUMBER: 2 312 31 A F F E C T E D -- NUMBER EXAMINED: 2 2 2 0 2 31 2 31 2 31 2 00 0 31 2 31 31 31 10 21 000227 91 TABLE 17 Incidence of Microscopic Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS TABLE INCLUDES: SEX=ALL;GROUP=ALL;WEEKS=ALL DEATH=T;FIND=ALL;SUBSET=ALL ORGAN AND FINDING DESCRIPTION ** TOP OF LIST ** EYE (EY) ........................................... BONE, FEMUR (FE) .................................. MARROW, FEMUR (FM) ................................ KIDNEY (KD) ........................................ --INFILTRATE, LYMPHOPLASMACYTIC LUNG (LU) .......................................... LIVER (LI) ......................................... --INFILTRATE, LYMPHOPLASMACYTIC --INFLAMMATION, LYMPHOHISTIOCYTIC -- LIPIDOSIS, SUBCAPSULAR (TENSION LIPIDOSIS) -- N UM BER -0 F-AN SEX: --MALE--- -- FEMALE--- GROUP: -l- -2- -3- -1- -2- -3- NUMBER: 2 3 1 2 3 1 2 312 31 NOT REMARKABLE: 2 3 1 2 3 1 2 31231 NOT REMARKABLE: 2 3 1 2 3 1 0 10 12 1 NOT REMARKABLE: 0 1 0 1 2 1 231231 NOT REMARKABLE: 2 2 1 1 1 1 010120 2 3 12 31 NOT REMARKABLE: 2 3 1 2 3 1 2 3 12 3 1 NOT REMARKABLE: 2 2 1 0 1 1 0 10 120 000100 000 100 Covance 6329-222 3M T-6295.6 PAGE: 1 A F F E C T E D -- 000228 92 0 1 CC 000229 TABLE 17 Incidence of Microscopic Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS TABLE INCLUDES: SEX=ALL;GROUP=ALL;WEEKS=ALL DEATH=T;FIND=ALL;SUBSET-ALL ORGAN AND FINDING DESCRIPTION SPLEEN (SP) ................. PANCREAS (PA) .............. ADRENAL, CORTEX (AC) ....... --HEMORRHAGE --THROMBUS ADRENAL, MEDULLA (MA) ...... . THYMUS (TH) ................. TESTIS (TE) ................. -- IMMATURE ** END OF LIST ** -- NUMBE F-AN SEX: --MALE- -- -- FEMALE- -- GROUP: - 1 - -2- -3- -l- -2- -3- NUMBER: 2 3 1 2 3 1 NUMBER EXAMINED: 2 3 1 2 3 1 NOT REMARKABLE: 2 3 1 2 3 1 NUMBER EXAMINED: 2 3 1 2 3 1 NOT REMARKABLE: 2 3 1 2 3 1 NUMBER EXAMINED: 2 3 1 2 3 1 NOT REMARKABLE: 2 3 1 2 2 1 0000 10 000 0 10 NUMBER EXAMINED: 2 3 1 2 3 1 NOT REMARKABLE: 2 3 1 2 3 1 NUMBER EXAMINED: 2 3 1 2 3 1 NOT REMARKABLE: 2 3 1 2 3 1 NUMBER EXAMINED: 2 3 1 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 2310 0 0 Covance 6329-222 3M T-6295.6 PAGE: 2 A F F E C T E D -- 93 APPENDIX 1 Protocol Deviations Protocol Protocol Amendment No. 1 Protocol Amendment No. 2 Protocol Amendment No. 3 Protocol Amendment No. 4 Material Safety Data Sheet Covance 6329-222 3M T-6295.6 000230 94 Covance 6329-222 3M T-6295.6 Protocol Deviations Protocol. Group Designations and Dosage Levels. Dose levels were 0, 0.02, and 2.0 mg/kg/day. The control group (Group 1) was to receive an equivalent amount o f lactose in gelatin capsules as the total material administered to Group 3. The total material was 20.0 mg/kg/day for each group; however, the low- and high-dose groups were to receive test material triturated with lactose (1:999, w:w, and 1:9, w:w, respectively). Actual Procedure. Because the capsule broke during dose administration, the following animals may not have received the full amount o f dose preparation: Animal No. 105350 (Group 1 male) on Day 1; Animal No. 105367 (Group 1 female) on Days 2, 7, and 14; Animal No. 105344 (Group 2 male) on Day 7; and Animal No. 105369 (Group 3 female) on Day 2. Protocol. Dose Analysis. Stability. "One set o f samples (approximately 1 g each) will be taken from the low- and high-dose test material/lactose preparations at the end o f the in-life phase and analyzed for test material content." Actual Procedure. Two reserve samples (10 g) were shipped to the sponsor for stability analysis. The samples for stability analysis (1 g) were retained as reserve samples, until shipped to the Sponsor on March 19, 2002. Protocol. Observation o f Animals. Rectal Body Temperatures. "Beginning 2 weeks before initiation of treatment, rectal body temperatures will be taken at approximately 11:00 on two days/week (Monday and Thursday)." Actual Procedure. Rectal body temperatures were recorded between 12:50 and 13:30 on Day -14 and between 08:17 and 08:40 on Day -10. Protocol. Serum PFOS Level Determination. Frequency. "Before initiation o f treatment (Day -7 or -6), and prior to treatment on Days 2 (approximately 24 hours after the first dose), 7, 14, and 29." 000231 95 Protocol Deviations (Continued) Covance 6329-222 3M T-6295.6 Actual Procedure. On Day 2, incorrect blood collection tubes were used for the collection o f blood for clinical chemistry test; therefore, blood collected for serum PFOS level determinations was used for clinical chemistry tests. Remaining serum from clinical chemistry tests was used as the Day 2 sample for serum PFOS level determinations. In addition, whole blood (approximately 2 mL) was collected from a femoral vein of each animal before treatment on Day 3 (approximately 24 hours after the second dose). Protocol. Additional Blood Collection. "At scheduled and unscheduled necropsies, blood (as much as possible, up to 20 mL) will be collected from animals at the time of exsanguination." Actual Procedure. Additional blood was not collected from Animal Nos. 105364 (Group 2 female), 105365 (Group 2 female), and 105369 (Group 3 female). Protocol. Postmortem Procedures. Tissue Preservation. "The following tissues (when present) from each animal will be preserved in 10% neutral-buffered formalin, unless otherwise specified, for possible future microscopic examination:" Actual Procedure. Femoral bone marrow from Animal Nos. 105349 and 105350 (Group 1 males), 105344 and 105348 (Group 2 males), 105345 (Group 3 male), 105366 (Group 1 female), and 105364 (Group 2 female) were insufficient; therefore, these tissues were not examined microscopically. This tissue is listed with the appropriate comments in the pathology data sheets for each individual animal. Summary tables do not include it as having been examined. These deviations are not expected to have affected the results o f the study. 000232 COVA1tfcl> THE DEVELOPMENT SERVICES COMPANY Sponsor: 3M St. Paul, Minnesota PROTOCOL Study Title: 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Date: April 17, 1998 Performing Laboratory: Covancc Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704-2595 Laboratory Study Identification: Proposal No. 90545A Covance 6329-222 000233 97 Covance 6329-222 _________ Page 2 Study 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Purpose To provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study and to assess the effect of the test material on critical enzyme levels, hormones, and other selected biochemical parameters Sponsor 3M Toxicology Services Building 220-2E-02, 3M Center St. Paul, Minnesota 55144-1000 Study Monitor Andrew M. Seacat, PhD 3M Telephone No.: 612.575.3161 Facsimile No.: 612.733.1773 Alternate Study Monitor Paul Lieder, PhD, DABT 3M Telephone No.: 612.737.2678 Facsimile No.: 612.733.1733 Study Location Covance Laboratories Inc. 3301 Kinsman Boulevard Madison. Wisconsin 53704-2595 Mailing Address: PO Box 7545 Madison, Wisconsin 53707-7545 000234 98 Study Director Peter J. Thomford, PhD Covance Laboratories Inc. Telephone No.: 608.241.7207 Facsimile No.: 608.242.2736 Covance 6329-222 _________ Page 3 Study Toxicologist Dale Aldridge, BS Covance Laboratories Inc. Proposed Study Timetable In-Life Start Date: April 23, 1998 In-Life End Date: May 22, 1998 Audited Draft Report Date: September 3, 1998 Regulatory Compliance This study will be conducted in compliance with the Environmental Protection Agency Good Laboratory Practice Regulations as set forth in Title 40 of the US Code of Federal Regulations, Part 792, issued November 29, 1983 (effective December 29, 1983), and with any applicable amendments. Animal Care and Use Statement All procedures in this protocol are in compliance with the Animal Welfare Act Regulations, 9 CFR 1-4. In the opinion of the Sponsor and study director, the study does not unnecessarily duplicate any previous work. Quality Assurance The protocol, study conduct, and final report will be audited by the Covance Quality Assurance Unit (QAU). The proliferation cell nuclear antigen evaluation data and report will be audited by the QAU of Pathology Associates International. The dose analysis and plasma and liver PFOS analysis and report will be audited by the QAU of 3M Environmental Laboratories. The blood hormone determinations and report will be audited by the QAU of AniLytics Inc. 000235 99 Covance 6329-222 ______________________________________________________ _________________ Page 4 Test Material Identification Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) Lot Number The lot number will be maintained in the raw data. Purity Responsibility of the Sponsor Stability Responsibility of the Sponsor Storage Conditions At room temperature Characteristics Information on synthesis methods, composition, or other characteristics that define the test material is on file with the Sponsor. Vehicle Identification Lactose Lot Number The lot number will be maintained in the raw data. Purity On file with the manufacturer Stability On file with the manufacturer 000236 100 Storage Conditions At room temperature Covance 6329-222 ___________Pa8e5 Gelatin Capsules Capsules (Size No. 2) obtained from Torpac, Inc. (Fairfield, New Jersey); lot number will be supplied by the manufacturer. Gelatin capsules will be stored at room temperature. A copy of the Certificate of Analysis provided by the manufacturer will be maintained in the data. Reserve (Archive) Samples A reserve sample of each lot of test material, vehicle, and each test material/lactose trituration (10 g each) will be taken and stored at room temperature. These samples will be transferred to the Sponsor after completion of the in-life phase. Disposition of Test Material Any remaining test material will be retained at Covance for use in possible future studies. Animals Species Cynomolgus monkey (purpose-bred) Source Covance Research Products Inc., Alice, Texas Age at Initiation of Treatment Young adult/adult Weight at Initiation of Treatment Approximately 2 to 4 kg Number and Sex Six males and six females Identification Collar tag 000237 101 Husbandry Covance 6329-222 ----------------- P*ge .6 Housing Individual; animals will be housed in suspended, stainless steel cages. Diet Certified primate diet (#8726C, Harlan Teklad) once or twice daily. The diet is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Results of specified nutrient and contaminant analyses are on file at Covance-Madison. Fruit, vegetables, or other supplements may be provided but will not require analysis. Water Ad libitum. Samples of the water arc routinely analyzed for specified microorganisms and environmental contaminants. The results are on file at Covance-Madison. Contaminants There are no known contaminants in the diet or water at levels that might interfere with this study. Environment Environmental controls for the animal room will be set to maintain 18 to 29C, a relative humidity of 30 to 70%, and a 12-hour light/12-hour dark cycle. Acclimation Minimum of 4 weeks Randomization Animals will be weighed, stratified by body weight, and allocated to the number of blocks equal to the number of animals to be selected for each group. Animals in each block will then be assigned to groups using computer-generated random numbers. 000238 102 Covance 6329-222 _________ Page 7 Justification PFOS is a known hepatic peroxisome proliferator (PP) in the rat. When exposed to PP, nonhuman primates (such as the cynomolgus monkey) respond similarly to humans (i.e., low to no hepatic response) and therefore are an appropriate human surrogate species. Group Designations and Dosage Levels Group Dose Level (mg/kg/day) Total Material Dose Number of Animals Level (mg/kg/day) Males Females 1 (control) 2 (low-dose) 3 (high-dose) 0a 0.02 2.0 20.0* 20.0b 20.0e 2 3 1 2 3 1 a The control group (Group 1) will receive an equivalent amount of lactose in gelatin capsules as the total material administered to Group 3. b The low-dose (Group 2) will receive the test material triturated with lactose (1:999, w:w). c The high-dose (Group 3) will receive the test material triturated with lactose (1:9, w:w). Dosing Procedures Method of Administration Orally by gelatin capsules, daily (7 days/week) for 28 days Reason for Dosing Route To compare with data from previous toxicology studies using the oral route, which is the most likely route of exposure Dose Preparation Capsules will be prepared at least twice weekly. The size and number of capsules will depend on the physical characteristics of the lest material, the dose level, and the weight of the monkey. Individual daily doses will be based on the most recent individual body weights, with the exception of body weight collection days when the previous body weight will be used. Dose levels will be based on the vehicle as 0002o9 103 Covance 6329-222 __________ PageS supplied for Group 1. For the Groups 2 and 3 dose preparations, the test material will be triturated with lactose (1:999 and 1:9, w:w, respectively) once before initiation of treatment. Trituration with lactose is necessary to facilitate capsule preparation. All dose preparations will be stored at room temperature until dosed. Dose Analysis Dose analyses will be done by the Sponsor. Homogeneity Samples (approximately 1 g each) will be collected from the top, middle, and bottom of the test material/lactose preparations for the low- and high-dose groups and analyzed for test material content. All samples will be stored at room temperature until analyzed. Stability Homogeneity samples collected from the middle of the preparations will be used for the prestudy stability analysis. One set of samples (approximately 1 g each) will be taken from the low- and high-dose test material/lactose preparations at the end of the in-life phase and analyzed for test material content. Sample Shipping Samples will be shipped under ambient conditions to: Kris J. Hansen, PhD 3M E.T. & S Bldg. 2-3E-09 935 Bush Avenue St. Paul, Minnesota 55106 Telephone: 612.778.6018 Facsimile: 612.778.6176 Kris J. Hansen or her alternate will be notified regarding the shipment of the samples. Analysis of for test material content will be done on the samples. Results will be provided for inclusion in the final report. 000240 104 Observation of Animals Covancc 6329-222 __________Page 9 Clinical Observations Each animal will be observed twice daily (a.m. and p.m.) for mortality and moribundity; findings will be recorded as they are observed. Each animal will be observed daily and food consumption will be assessed qualitatively; abnormal findings will be recorded. Once weekly, each animal will be observed; abnormal findings, or an indication that the animal is normal will be recorded. During treatment, each animal will be observed for signs of poor health or abnormal behavior approximately 30, 60, and 90 minutes postdose. Additional findings will be recorded as they are observed. Body Weights Beginning two weeks before initiation of treatment, twice weekly (Monday and Thursday), on the first day of treatment, and twice weekly thereafter. An additional body weight will be recorded on Day -1 for the Day 1 dose calculations. Rectal Body Temperatures Using nonanesthetized animals. Beginning 2 weeks before initiation of treatment, rectal body temperatures will be taken at approximately 11:0() on two days/week (Monday and Thursday). Clinical Pathology Frequency Unscheduled Collections When possible, blood will be collected for clinical pathology tests from animals sacrificed at unscheduled intervals 000241 105 Covance 6329-222 ________ Page 10 Scheduled Collections Hematology and clinical chemistry will be done once before initiation of treatment (Day -7 or -6) and on Day 29. Clinical chemistry will also be done prior to the daily dose on Days 2, 7, and 14. Tests Method of Collection Animals will be fasted overnight; blood will be collected from a femoral vein. Anticoagulant will be potassium EDTA for hematology tests. No anticoagulant is used for clinical chemistry tests. Hematology red blood cell (erythrocyte) count hemoglobin hematocrit mean corpuscular volume mean corpuscular hemoglobin mean corpuscular hemoglobin concentration platelet count white blood cell (leukocyte) count differential blood cell count blood cell morphology reticulocyte count Clinical Chemistry glucose urea nitrogen creatinine total protein albumin globulin total bilirubin direct bilirubin (if total bilirubin is greater than 2.0 mg/dL) cholesterol triglycerides alanine aminotransferase alkaline phosphatase aspartate aminotransferase gamma glutamyltransferase sorbitol dehydrogenase creatine kinase calcium inorganic phosphorus sodium potassium chloride serum bile acids amylase lipase pancreatic-specilic amylase 000242 106 Covance 6329-222 _____________________________________________________ __________________Page 11 Blood Hormone Determination Frequency Before initiation of treatment (Day -7 or -6) and prior to treatment on Day 29; blood will be collected between 7:00 a.m. and 9:00 a.m. Number of Animals All Method of Collection Animals will be fasted overnight; blood will be collected from a femoral vein. Approximately 5 mL of blood will be collected without anticoagulant and allowed to clot for serum samples. Sample Handling Samples for serum will be centrifuged within 1 hour after collection, and serum will be harvested. Serum will be stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to: Dr. Das AniLytics Inc. 200 Girard Street, Suite 200 Gaithersburg, Maryland 20877 Telephone No.: 301.921.0168 Facsimile No.: 301.977.0433 AniLytics Inc. will be notified regarding shipment of samples. Tests Samples will be analyzed by AniLytics Inc., for estradiol, estrone, estriol, thyroid stimulating hormone, triiodothyronine (T3), and thyroxin (T4). Serum PFOS Level Determination Frequency Before initiation of treatment (Day -7 or -6), and prior to treatment on Days 2 (approximately 24 hours after the first dose), 7, 14, and 29 000243 107 Number of Animals All Covance 6329-222 ________ Page 12 Method of Collection Animals will be fasted overnight; blood (approximately 2 mL) will be collected from a femoral vein without an anticoagulant. Sample Handling Samples will be centrifuged within 1 hour after collection, and serum will be harvested. Serum samples will be stored in a freezer set to maintain -60 to -80C. Sample Shipping Serum samples will be packed on dry ice and shipped to: Kris J. Hansen, PhD 3M E.T. & S Bldg. 2-3E-09 935 Bush Avenue St. Paul, Minnesota 55106 Telephone: 612.778.6018 Facsimile: 612.778.6176 Kris J. Hansen or her alternate will be notified regarding the shipment of the samples. Analysis of serum for PFOS will be done on the samples. Results will be provided for inclusion in the final report. Additional Blood Collection At scheduled and unscheduled necropsies, blood (as much as possible, up to 20 mL) will be collected from animals at the time of exsanguination. Blood from each animal will be transferred into containers (approximately 5 mL each) and stored in a freezer set to maintain -60 to -80C until shipped to the Sponsor for possible future analysis. 000244 108 Termination Covance 6329-222 ________ Page 13 Unscheduled Sacrifices and Deaths Necropsies will be done. Animals to be sacrificed will be anesthetized with sodium pentobarbital, weighed, bled for required tests, and exsanguinated. Scheduled Sacrifice After at least 4 weeks of treatment, all surviving animals will be fasted overnight, then anesthetized with sodium pentobarbital, weighed, bled for required tests, exsanguinated, and necropsied. Postmortem Procedures Necropsy The necropsy will include examination of: all orifices cranial cavity external surface of the brain; the external surface of the spinal cord and cut surfaces of the brain and spinal cord will be examined whenever tissue trimming is performed cervical tissues and organs thoracic, abdominal, and pelvic cavities and viscera external surface of the body nasal cavity and paranasal sinuses Palmitoyl CoA Oxidase Determinations A sample of the right lateral lobe of liver will be collected from each animal at the scheduled sacrifice. The sample will be flash-frozen in liquid nitrogen, and stored in a freezer set to maintain -60 to -80C until analyzed for palmitoyl CoA oxidase activity. Cell Proliferation Evaluation Representative samples of the liver, testes, and pancreas will be collected and preserved in zinc formalin. After fixation, samples for proliferation cell nuclear antigen (PCNA) evaluation will be embedded in paraffin and shipped to: 000245 109 Sandra R. Eldridge, PhD Pathology Associates International 15 Worman's Mill Court Suite I Frederick, Maryland 21701 Telephone: 301.663.1644 ext. 2201 Facsimile: 301.663.8994 Covance 6329-222 ________ Page 14 Pathology Associates International will be notified regarding shipment of samples. PCNA evaluation will be done on the samples. Results will be provided for inclusion in the final report. Liver PFOS Determination A sample of liver (any non-formalin treated liver remaining after sampling for histopathology) will be collected from each animal at the scheduled sacrifice, weighed, flash-frozen in liquid nitrogen, and stored in a freezer set to maintain -60 to -80C until shipped with plasma samples to 3M (Kris Hansen) for analysis. Analysis of liver for PFOS will be done on the samples. Results will be provided for inclusion in the final report. Tissue Preservation The following tissues (when present) from each animal will be preserved in 10% neutral-buffered formalin, unless otherwise specified, for possible future microscopic examination: adrenal (2) aorta brain cecum cervix colon duodenum epididymis (2) esophagus eye [(2) to be preserved in Davidson's fixative] femur with bone marrow (articular surface of the distal end) gallbladder mesenteric lymph node pancreas pituitary prostate rectum salivary gland [mandibular (2)] sciatic nerve seminal vesicle (2) skeletal muscle (thigh) skin spinal cord (cervical, thoracic, and lumbar) spleen sternum with bone marrow 000246 no heart ileum jejunum kidney (2) lesions liver lung mammary gland (females only) ovary (2) Covance 6329-222 _________Page 15 stomach testis (2) thymus thyroid (2) with parathyroid trachea urinary bladder uterus vagina Bone Marrow Smear From the sternum of each animal at unscheduled and scheduled sacrifices (made and held for possible future examination) Histopathology The adrenals, eye, femoral bone marrow, lung, liver, kidney, pancreas, spleen, testes, and thymus from each animal will be embedded in paraffin, sectioned, stained with hematoxylin and eosin, and examined microscopically. Reports A draft report that includes the following information will be prepared and submitted: Experimental Design and Methods Results mortality clinical observations body weights food consumption rectal body temperatures clinical pathology results palmitoyl CoA oxidase activities hormone analyses (provided by AniLytics) dose analyses (provided by 3M) plasma and liver PFOS levels (provided by 3M) macroscopic observations microscopic observations cell proliferation evaluations (provided by Pathology Associates International) dose analyses (provided by 3M) 000247 ill Covancc 6329-222 ________ Page 16 Statistical Evaluation Levene's test will be done to test for variance homogeneity. In the case of heterogeneity of variance at p 0.05, transformations will be used to stabilize the variance. Comparison tests will take variance heterogeneity into consideration. One-way analysis of variance (ANOVA) will be used (if applicable) to analyze initial body weights, rectal body temperature, palmitoyl CoA oxidase activities, and continuous clinical pathology values. If the ANOVA is significant, Dunnett's t-test will be used for control versus treated group comparisons. One-way analysis of covariance (ANCOVA) will be used to analyze body weights, with initial body weights as the covariate. If the ANCOVA is significant, covariate-adjusted means will be used for control versus treated group comparisons. Group comparisons (Group 2 versus Group 1) will be evaluated at the 5.0% two-tailed probability level. Only data collected on or after the first day of treatment will be analyzed statistically. At the end of 1 year after issuance of the audited draft report, if no requested revisions or instructions to finalize have been communicated by the Sponsor, then the audited draft report will be considered 'final' and issued as the final report, signed by the study director, and submitted to the Sponsor. Any modifications or changes to the audited draft report requested 1 year after issuance will be performed at additional cost to the Sponsor. Record Retention All raw data, documentation, records, protocol, specimens, and final report generated as a result of this study will be archived in the storage facilities of Covance for a period of 1 year following submission of the final report to the Sponsor. One year after submission of the final report, all of the aforementioned materials will be sent to the Sponsor, and a return fee will be charged. The Sponsor may elect to have the materials retained in the Covance archives for an additional period of time, and Covance will charge a storage fee. If the Sponsor chooses to have Covance dispose of the materials, a disposal fee will be charged. All raw data stored on magnetic media will be retained by Covance. 000248 112 protocol and protocol amendments dose preparation records in-life records animal receipt acclimation animal room maintenance randomizations dose administration clinical observations body weights food consumption sample collection clinical pathology records anatomical pathology records statistical analyses study correspondence tissue specimens (wet and in paraffin) blood and tissue slides final report (original signed copy) Covance 6329-222 ________ Page 17 The following supporting records will be retained at Covance-Madison but will not be archived with the study data. feed analysis records water analysis records animal room environment records refrigerator and freezer temperature records room temperature records for test material storage instrument calibration and maintenance records. 000249 PROTOCOL APPROVAL Covance 6329-222 ________ Page 8 Date oooso c o v / n rc I> THE DEVELOPMENT SERVICES COMPANY PROTOCOL AMENDMENT NO. 1 Covance 6329-222 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Sponsor: Study Monitor: Testing Facility: Study Director: 3M, St. Paul, Minnesota Andrew M. Seacat, PhD Covance Laboratories Inc., Madison, Wisconsin Peter J. Thomford, PhD This amendment modifies the following portions of the protocol: Effective April 17,1998 1. Page 7, Dosing Procedures, Method of Administration. To more precisely define the duration of treatment, delete the text in this section replace with the following: Orally by gelatin capsules, daily (7 days/week) for at least 28 days 2. Page 11, Blood Hormone Determination, Sample Handling, Sentence 2. To modify the sample handling method, delete this sentence and replace with the following: Serum will be divided into two approximately equal aliquots and stored in a freezer set to maintain -60 to -80C until packed on dry ice and shipped to: 00051 115 Covance 6329-222 Protocol Amendment No. 1 _______________ Page 2 3. Page 14, Postmortem Procedures, Tissue Preservation. To correct an error in the protocol, delete "mammary gland (females only)" and replace with the following: mammary gland Effective April 28,1998 4. Page 2, Alternate Study Monitor. To change the alternate study monitor, delete the text in this section and replace with the following: Marvin T. Case, DVM, PhD 3M Toxicology Services Telephone No.: 612.733.5180 Facsimile No.: 612.733.1773 AMENDMENT APPROVAL Date 3M 000252 COV/rttiT' TM1 O W tL O m iH T SH V ICSS COMPANY PROTOCOL AMENDMENT NO. 2 Covance 6329-222 4-Week Capsule Toxicity Study with Perfhiorooctane Sulfonic Add Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Sponsor: Study Monitor Testing Facility: Study Director 3M, St Paul, Minnesota Andrew M. Seacat, PhD Covance Laboratories Inc., Madison, Wisconsin Peter J. Thomford, PhD This amendment modifies the following portions of the protocol: Effective May 15,1998 1. Page 13, Termination, Unscheduled Sacrifices and Deaths. To change the anesthetic to be used before exsanguination because the barbiturate may interfere with tissue analyses, delete the text in this section and replace with the following: Necropsies will be done. Animals to be sacrificed will be anesthetized with ketamine and xylazine, weighed, bled for required tests, and exsanguinated. 2. Page 13, Termination, Scheduled Sacrifice. To change the anesthetic to be used before exsanguination because the barbiturate may interfere with tissue analyses, delete the text in this section and replace with the following: After at least 4 weeks of treatment, all surviving animals will be fasted overnight, then anesthetized with ketamine and xylazine, weighed, bled for required tests, exsanguinated, and necropsied. 000253 117 000254 AMENDMENT APPROVAL Covane* 6329-222 Protocol Amendment No. 2 ______________Pa*e2 Andrew M. Seacat, PhD/^ Study Monitor 3M Date &> /fto A r k ' ^ fy lZ rc L ^ ' 7^ y j. r d u " ' * k/C 000255 PROTOCOL AMENDMENT NO. 3 Covance 6329-222 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Sponsor: Study Monitor: Testing Facility: Study Director: 3M, St. Paul, Minnesota Andrew M. Seacat, PhD Covance Laboratories Inc., Madison, Wisconsin Peter J. Thomford, PhD_____________________ This amendment modifies the following portions of the protocol. Effective April 17,1998 1. Page 5, Vehicle, Storage Conditions. To include the solvent used to dissolve the test material before mixing with the lactose, add the following after this section. Solvent Identification Acetone Lot Numbers The lot numbers will be maintained in the raw data. Purity On file with the manufacturer Stability On file with the manufacturer Storage Conditions At room temperature Characteristics Information on synthesis methods, composition, or other characteristics that define the solvent is on file with the manufacturer. 000256 120 Covance 6329-222 Protocol Amendment No. 3 ______________________________________________________________________________ Page 2 2. Page 7, Dosing Procedures, Dose Preparation, Sentence 5. To include the use o f acetone in the preparation of the doses, delete this sentence and replace with the following. For the Group 2 and 3 dose preparations, the test material will be dissolved in acetone and triturated with lactose (1:999 and 1:9, w:w, respectively) once before initiation o f treatment. Effective April 17,1998 and April 26,1999 3. Page 16, Record Retention. To correct an omission in the protocol and to specify the record retention requirements for Ani Lytics Inc. and 3M E.T. & S (effective April 17, 1998) and to clarify the requirement for Pathology Associates International (effective April 26, 1999), add the following. Within 1 year after submission of the final report, all o f the aforementioned materials (as appropriate) from Ani Lytics Inc. and 3M E.T. & S will be sent to the Sponsor (Andrew Seacat, PhD, 3M). Pathology Associates International (PAI) is responsible for the maintenance o f any raw data or specimens produced by PAI. Effective December 15,1998 4. Page 13, Postmortem Procedures, Cell Proliferation Evaluation, Paragraph 3, Sentence 2. To include evaluation o f slides stained with hematoxylin and eosin in the PCNA evaluation, delete this sentence and replace with the following. PCNA evaluation (including examination of slides stained with hematoxylin and eosin) will be done on the samples. 000257 121 AMENDMENT APPROVAL Covance 6329-222 Protocol Amendment No. 3 _________________ Page 3 Andrew M. Seacat, PhD Study Monitor 3M 000258 COV/HCl> THE DEVELOPMENT SERVICES COMPANY PROTOCOL AMENDMENT NO. 4 Covance 6329-222 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Sponsor: 3M, St. Paul, Minnesota Study Monitor: Andrew M. Seacat, PhD Testing Facility: Covance Laboratories Inc., Madison, Wisconsin Study Director:_____ Peter J. Thomford, PhD___________________ This amendment modifies the following portions of the protocol. Effective July 18,2000 1. Page 12, Serum PFOS Level Determination, Sample Shipping, Paragraph 3, Sentence 2. To reflect the sponsor's decision to report the results of analysis of tissues for compound levels separately, delete this sentence and replace with the following. Results will be reported separately by the sponsor. 2. Page 14, Liver PFOS Determination, Sentence 3. To reflect the sponsor's decision to report the results of analysis of tissues for compound levels separately, delete this sentence and replace with the following. Results will be reported separately by the sponsor. 3. Page 15, Reports, Results. To reflect the sponsor's decision to report the results of analysis of tissues for compound levels separately, delete the following. plasma and liver PFOS levels (provided by 3M) 00059 123 AMENDMENT APPROVAL Covance 6329-222 Protocol Amendment No. 4 _______________ Page 2 Study Monitor 3M 0002C0 Covance 6329-222 3M T-6295.6 MATERIAL SAFETY DATA SHEET 3M 3M Center St. Paul, Minnesota 55144-1000 (612) 733-1110 Copyright, 1997, Minnesota Mining and Manufacturing Company. All rights reserved. Copying and/or downloading of this information for the purpose of properly utilizing 3M products is allowed provided that: 1 ) the information is copied in full with no changes unless prior agreement is obtained from 3M, and 2) neither the copy nor the original is resold or otherwise distributed with the intention of earning a profit thereon. DIVISION: SPECIALTY CHEMICALS DIVISION TRADE NAME: FC-95 FLUORAD Brand Fluorochemical Surfactant ID NUMBER/U.P.C. : 98-0207-0103-7 00-51135-09054-1 98-0207-0104-5 98-0211-0888-5 00-51135-09362-7 98-0211-3916-1 ZF-0002-1044-1 ISSUED: February 19, 1997 SUPERSEDES: August 23, 1996 DOCUMENT: 10-3796-9 00-51135-09055-8 00-51135-02311-2 . 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:................ N/A VAPOR PRESSURE:............... N/A VAPOR DENSITY:................ N/A EVAPORATION RATE:............. N/A SOLUBILITY IN WATER:.......... slight SPECIFIC GRAVITY:............. ca. 0.6 Water=1 (Bulk) PERCENT VOLATILE:............. 0 % pH:........................... 7 - 8 (0.1% Aqueous) VISCOSITY:.................... N/D cTING POINT:................ N/D t At-PEARANCE AND ODOR: Light colored, free flowing powder. 000261 125 Covance 6329-222 3MT-6295.6 /SOS: FC-95 FLUORAO Brand Fluorochemical Surfactant February 19, 1997 PAq E 2 3. FIRE AND EXPLOSION HAZARD DATA FLASH POINT:................... FLAMMABLE LIMITS - LEL:..... FLAMMABLE LIMITS - UEL:..... AUTOIGNITION TEMPERATURE:..... None N/A N/A N/A EXTINGUISHING MEDIA: Hater, Carbon dioxide, Dry chemical, Foam SPECIAL FIRE FIGHTING PROCEDURES: Hear full protective clothing, including helmet, self-contained, positive pressure or pressure demand breathing apparatus, bunker coat and pants, bands around arms, waist and legs, face mask, and protective covering for exposed areas of the head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See Hazardous Decomposition section for products of combustion. *. REACTIVITY DATA STABILITY: Stable INCOMPATIBILITY - MATERIALS/CONDITIONS TO AVOID: Not applicable. HAZARDOUS POLYMERIZATION: Hazardous polymerization will not occur. HAZARDOUS DECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Sulfur, Hydrogen Fluoride, Toxic Vapors, Gases or Particulates. 5. ENVIRONMENTAL INFORMATION SPILL RESPONSE: Observe precautions from other sections. Vacuum, use wet sweeping compound or water to avoid dusting. CAUTION! A vacuum cleaner could be an ignition source. Clean up residue with water. Place in an approved metal container. Seal the container. "ECOMMENDED DISPOSAL: 1o 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 000262 126 Covance 6329-222 3M T-6295.6 FC-J95 FLUORAD Brand Fluorochemical Surfactant uary 19, 1997 PAGE 3 5. ENVIRONMENTAL INFORMATION (continued) accept chemical Haste. ENVIRONMENTAL DATA: 96-Hr. Aquatic Fish LC50, Fathead Minnow(Pimephales promelas)=38 mg/1, Bluegill Sunfish(Lepomis macrochirus)=68 mg/1, Rainbow Trout(Salmo gairdneri)=11 ag/1; 48-Hr. EC50, Daphnia Magna = 50 mg/1; C00=.004 g/g; BOD20 = Nil. REGULATORY INFORMATION: Volatile Organic Compounds: N/A. VOC Less H20 & Exempt Solvents: N/A. Since regulations vary, consult applicable regulations or authorities before disposal. U.S. EPA Hazardous Waste Number = None (Not U.S. EPA Hazardous). This product complies with the chemical registration requirements of TSCA, EINECS, CDSL, AICS, MITI and Korea. -CRA HAZARD CLASS: IRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes 6. SUGGESTED FIRST AID EYE CONTACT: Immediately flush eyes with large amounts of water for at least 15 minutes. Get immediate medical attention. SKIN CONTACT: Immediately flush skin with large amounts of water. Remove contaminated clothing. If irritation persists, call a physician. Wash contaminated clothing before reuse. INHALATION: If signs/symptoms occur, remove person to fresh air. If signs/symptoms continue, call a physician. IF SWALLOWED: Drink two glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION t-._ PROTECTION: Avoid eye contact. Wear vented goggles. 000263 127 Covance 6329-222 3M T-6295.6 ,<SDS: FC-95 FLUORAD Brand Fluorochemical Surfactant e<*bruary 19, 1997 4. 7. PRECAUTIONARY INFORMATION (continued) SKIN PROTECTION: Avoid skin contact. Wear appropriate gloves when handling this material. A pair of gloves made from the following material(s) are recommended: butyl rubber. Use one or more of the following personal protection items as necessary to prevent skin contact: head covering, coveralls. Protective garments (other than gloves) should be made of either of the following materials: polyethylene/polyvinylidene chloride (Saranex). RECOMMENDED VENTILATION: Use with appropriate local exhaust ventilation. Use in a wellventilated area. Provide sufficient ventilation to maintain emissions below recommended exposure limits. If exhaust ventilation is not adequate, use appropriate respiratory protection. RESPIRATORY PROTECTION: Avoid breathing of dust. Select one of the following NIOSH approved respirators based on airborne concentration of contaminants and in accordance with OSHA regulations: half-mask dust and mist respirator, half-mask supplied air respirator, full-face dust and mist respirator, ull-face supplied air respirator. PREVENTION OF ACCIDENTAL INGESTION: Do not eat, drink or smoke when using this product. Wash exposed areas thoroughly with soap and water. Wash hands after handling and before eating. RECOMMENDED STORAGE: Keep container dry. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: Nonflammable. OTHER PRECAUTIONARY INFORMATION: No smoking: Smoking while using this product can result in contamination of the tobacco and/or smoke and lead to the formation of the hazardous decomposition products mentioned in section 4 of this MSDS. HMIS HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.) EXPOSURE LIMITS "3REDIENT VALUE UNIT TYPE AUTH SKIN POTASSIUM PERFLUOROALKYL SULFONATE... POTASSIUM PERFLUOROALKYL SULFONATE... POTASSIUM PERFLUOROALKYL SULFONATE... POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 0.1 0.1 0.1 MG/M3 MG/M3 MG/M3 MG/M3 128 TWA 3M Y TWA 3M Y TWA 3M Y TWA 3M . Y 000264 Covance 6329-222 3M T-6295.6 FC-95 FLUORAD Brand Fluorochemical Surfactant jary 19, 1997 EXPOSURE LIMITS (continued) PAGE 5 INGREDIENT POTASSIUM PERFLUOROALKYL SULFONATE... VALUE UNIT 0.1 MG/MS TYPE AUTH SKIN* TWA 3M Y * SKIN NOTATION: Listed substances indicated with `Y' under SKIN refer to the potential contribution to the overall exposure by the cutaneous route including mucous membrane and eye, either by airborne or, more particularly, by direct contact with the substance. Vehicles can alter skin absorption. SOURCE OF EXPOSURE LIMIT DATA: - 3M: 3M Recommended Exposure Guidelines 8. HEALTH HAZARD DATA EYE CONTACT: Mild Eye Irritation: signs/symptoms can include redness, swelling, pain, and tearing. SKIN CONTACT: Mild Skin Irritation (after prolonged or repeated contact): jigns/symptoms can include redness, swelling, and itching. May be absorbed through the skin and persist in the body for an extended time. INHALATION: May be harmful if inhaled. May be absorbed by inhalation and persist in the body for an extended time. Single overexposure, above recommended guidelines, may cause: Irritation (upper respiratory): signs/symptoms can include soreness of the nose and throat, coughing and sneezing. IF SWALLOWED: Ingestion is not a likely route of exposure to this product. Illness may result from a single swallowing of a moderate quantity of this material. May be harmful if swallowed. TAGENICITY: utagenicity assays indicate the product is not mutagenic. 000265 129 Covance 6329-222 3M T-6295.6 iSDS: FC-95 FLUORAD Brand Fluorochemical Surfactant February 19, 1997 PAQE 5 8 . HEALTH HAZARD DATA (continued) REPRODUCTIVE/DEVELOPMENTAL TOXINS: Not teratogenic in the rat at oral doses below maternally toxic levels. OTHER HEALTH HAZARD INFORMATION: A Product Toxicity Summary Sheet is available. SECTION CHANGE OATES PRECAUTIONARY INFO. SECTION CHANGED SINCE August 23, 1996 ISSUE Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately The information in this Material Safety Data Sheet (MSDS) is believed to be correct as of the date issued. 3M MAKES NO WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User is responsible for determining whether the 3M product is fit for a particular purpose and suitable for user's method of use or application. Given the variety of factors that can affect the use and application of a 3M product, some of which are uniquely within the user's knowledge and control, it is essential that the user evaluate the 3M product to determine whether it is fit for a "articular purpose and suitable for user's method of use or application. provides information in electronic form as a service to its customers. Due to the remote possibility that electronic transfer may have resulted in errors, omissions or alterations in this information, 3M makes no representations as to its completeness or accuracy. In addition, information obtained from a database may not be as current as the information in the MSDS available directly from 3M. 130 APPENDIX 2 Individual Animal Fate Data Individual Clinical Observations Individual Rectal Body Temperature Data (C) Covance 6329-222 3M T-6295.6 000267 APPENDIX 2 Individual Animal Fate Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE : 1 ANIMAL DOSE NUMBER GROUP SEX 105349 105350 105344 105347 105348 105345 105366 105367 105364 105365 105370 105369 1 MALE 1 MALE 2 MALE 2 MALE 2 MALE 3 MALE 1 FEMALE 1 FEMALE 2 FEMALE 2 FEMALE 2 FEMALE 3 FEMALE DEATH CODE T T T T T T T T T T T T TYPE OF DEATH SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED SCHEDULED DESCRIPTION OF DEATH TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE TERMINAL SACRIFICE DATE OF DAY OF DEATH STUDY 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 05/22/98 30 30 30 30 30 30 30 30 30 30 30 30 WEEK OF STUDY 5 5 5 5 5 5 5 5 5 5 5 5 000268 132 Appendix 2 Individual Clinical Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 PAGE: 1 ANIMAL DEATH WK OF NUMBER CODE DEATH CATEGORY GROUP: Ml DOSE: 0 MG/KG/DAY KEYWORD QUALIFIER DAYS 1-30 105349 T 5 NORMAL NO REMARKABLE OBSERVATIONS 105350 T 5 EXCRETION MUCOID FECES NORMAL NO REMARKABLE OBSERVATIONS 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP AM OBS 30MIN 90MIN UNSCHED DAY PM CHECK 60MIN DISPATCH iP- - - - - - 8P- - - - - - 15 P - - - - - - 22 P - - - - - - 29 P - - - - - - 30 P - - - - P " 12 - - - P - - - 1P- - - - - - 8P- - - - - - 15 P - - - - - - 22 P - - - - - - 29 P - - - - - - 30 P - - - - P- 000269 133 Appendix 2 Individual Clinical Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 PAGE: 2 ANIMAL DEATH WK OF NUMBER CODE DEATH CATEGORY GROUP: M2 DOSE: 0.02 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP KEYWORD AM OBS 30MIN 90MIN UNSCHED QUALIFIER DAYS 1-30 DAY PM CHECK 60MIN DISPATCH 105344 T 5 NORMAL NO REMARKABLE OBSERVATIONS 105347 T 5 DISCHARGE VOMITUS CONTAINING FOOD NORMAL NO REMARKABLE OBSERVATIONS 105348 T 5 NORMAL NO REMARKABLE OBSERVATIONS QUALITATIVE FOOD CONSUMPTION LOW 1P 8P 15 P 22 P 29 P 30 P 4 1P 8P 15 P 22 P 29 P 30 P 1P 8P 15 P 22 P 29 P 30 P 15 P P P P P 000270 134 Appendix 2 Individual Clinical Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 PAGE: 3 ANIMAL DEATH WK OF NUMBER CODE DEATH CATEGORY GROUP: M3 DOSE: 2 MG/KG/DAY KEYWORD QUALIFIER DAYS 1-30 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP AM OBS 30MIN 90MIN UNSCHED DAY PM CHECK 60MIN DISPATCH 105345 T 5 DISCHARGE VOMITUS CONTAINING FOOD DISCHARGE UNKNOWN SOURCE FOUND IN PAN RED IN COLOR NORMAL NO REMARKABLE OBSERVATIONS 24 P - - - - - - 15 P - - - - - _ 1 P- - - - - - 8P- - - - - - 22 P - - - - - - 29 P - - - - - - 30 P - - - - P- 000271 135 Appendix 2 Individual Clinical Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 PAGE: 4 ANIMAL DEATH WK OF NUMBER CODE DEATH CATEGORY GROUP: FI DOSE: 0 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP KEYWORD AM OBS 30MIN 90MIN UNSCHED QUALIFIER DAYS 1-30 DAY PM CHECK 60MIN DISPATCH 105366 T 5 EXCRETION NON-FORMED FECES NORMAL NO REMARKABLE OBSERVATIONS 105367 T 5 APPEARANCE SWOLLEN LIP(S) EXCRETION NON-FORMED FECES 23 P - P- - - - 27 P - - - - - - 29 P " " " - - - 1P"- -- 8P :: 15 P - - - - - - 22 P - - - - - - 30 P - - - - p- il 13 14 P 15 P - PPP - PPP - P- - - PPP - - 19 P 20 P 21 P 22 P 23 P 24 25 26 P - PPP PPPPPPPPP- ---- PPP- PPPPP- - 30 P - - - - P - 15 P 27 P - __ -- ___ --- 000272 136 Appendix 2 Individual Clinical Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 PAGE: 5 ANIMAL DEATH WK OF NUMBER CODE DEATH CATEGORY GROUP: FI DOSE: 0 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP KEYWORD AM OBS 30MIN 90MIN UNSCHED QUALIFIER DAYS 1-30 DAY PM CHECK 60MIN DISPATCH (CONTINUED FROM PREVIOUS PAGE) 105367 T 5 SKIN & PELAGE RED SKIN LIP(S) SCAB(S) MOUTH NORMAL NO REMARKABLE OBSERVATIONS QUALITATIVE FOOD CONSUMPTION LOW il 13 14 P 15 P 19 P 20 P 29 P 1P 8P 4P - - - - PPP- PPP- P- - - P PP - PPP- PPP- PPP- ------- ---- - _ - - c cc fo * cj 137 Appendix 2 Individual Clinical Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-62 95.6 PAGE: 6 ANIMAL DEATH WK OF NUMBER CODE DEATH CATEGORY GROUP: F2 DOSE: 0.02 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP KEYWORD AM OBS 30MIN 90MIN UNSCHED QUALIFIER DAYS 1-30 day PM CHECK 60MIN DISPATCH 105364 T 5 NORMAL NO REMARKABLE OBSERVATIONS 105365 T 5 DISCHARGE APPEARS TO BE MENSTRUATING NORMAL NO REMARKABLE OBSERVATIONS 105310 T 5 NORMAL NO REMARKABLE OBSERVATIONS 1P- - * - - - 8P- - - - - - 15 P - - - - - - 22 P - - - - - - 29 P - - - - - - 30 P - - - P- 29 P - - - - - - 30 P - - - - P- 1P - - - - - - 8P- - - - - - 15 P - - - - - - 22 P - - - - - - 1P- - - - - - 8P- - - - - - 15 P - - - - - - 22 P - - - - - - 29 P - - - - - - 30 P - - - - P- 000274 138 Appendix 2 Individual Clinical Observations 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS Covance 6329-222 3M T-6295.6 PAGE: 7 ANIMAL DEATH WK OF NUMBER CODE DEATH CATEGORY GROUP: F3 DOSE: 2 MG/KG/DAY 'C' - COMMENTS LISTED AT END OF OBSERVATIONS FOR SEX/GROUP KEYWORD AM OBS 30MIN 90MIN UNSCHED QUALIFIER DAYS 1-30 DAY PM CHECK 60MIN DISPATCH 105369 T ' 5 EXCRETION FEW FECES NORMAL NO REMARKABLE OBSERVATIONS QUALITATIVE FOOD CONSUMPTION LOW 17 P - - - - - - 1P- - - - - - 8P- - - - - - 15 P - - - - - - 22 P - - - - - - 29 P - - - - - - 30 P - - - - P- 26 P - - - - - - 000275 139 Appendix 2 Individual Rectal Body Temperature Data (C) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS ANIMAL DAY DAY DAY NUMBER -14 -10 -7 GROUP: MALE 1 - 0 MG/KG/DAY 105349 105350 40.0 39.6 40.2 39.7 40.2 39.4 GROUP: MALE 2 - 0.02 MG/KG/DAY 105344 105347 105349 39.0 39.8 38.7 39.0 39.7 39.0 38.4 39.8 39.8 GROUP: MALE 3 - 2 MG/KG/DAY 105345 39.6 39.5 39.1 GROUP: FEMALE 1 - 0 MG/KG/DAY 105366 105367 40.0 40.4 39.4 40.1 39.7 40.0 GROUP: FEMALE 2 - 0 . 0 2 MG/KG/DAY 105364 105365 105370 40.6 39.6 40.1 40.1 39.1 40.0 40.3 39.6 40.2 GROUP: FEMALE 3 - 2 MG/KG/DAY 105369 40.0 39.8 39.9 DAY -3 39.8 39.4 39.2 39.7 38.8 39.2 39.4 40.1 39.9 39.7 39.8 39.7 DAY 1 39.4 38.6 39.7 39.4 39.1 39.4 39.3 39.8 40.2 39.7 39.7 39.5 DAY 5 38.8 38.9 39.3 39.5 38.4 39.9 38.7 39.2 40.0 39.5 39.8 39.7 DAY 8 38.2 38.7 39.4 39.4 38.7 39.6 38.8 39.4 40.0 39.6 39.9 39.9 DAY 12 38.9 39.0 39.3 39.5 38.6 40.0 39.1 39.3 40.0 39.5 39.8 39.7 Covance 6329-222 3M T-6295.6 PAGE: 1 DAY 15 39.7 39.7 39.8 40.0 39.2 39.7 39.3 40.1 40.2 40.0 39.8 39.8 DAY 19 38.5 38.8 39.2 39.0 38.0 38.9 38.7 39.2 39.5 39.1 39.4 39.3 000276 140 000277 Appendix 2 Individual Rectal Body Temperature Data ("C) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS ANIMAL DAY DAY DAY NUMBER 22 26 29 GROUP: MALE 1 - 0 MG/KG/DAY 105349 105350 39.2 39.5 38.7 38.8 38.2 38.8 GROUP: MALE 2 - 0.02 MG/KG/DAY 105344 105347 105348 40.0 39.7 38.9 39.4 39.5 38.8 38.8 39.1 38.4 GROUP: MALE 3 - 2 MG/KG/DAY 105345 39.4 39.2 38.9 GROUP: FEMALE 1 - 0 MG/KG/DAY 105366 105367 39.3 39.9 38.7 39.2 38.7 38.9 GROUP: FEMALE 2 - 0.02 MG/KG/DAY 105364 105365 105370 40.1 39.6 39.8 39.8 39.5 39.6 39.3 38.7 39.4 GROUP: FEMALE 3 - 2 MG/KG/DAY 105369 39.4 39.1 39.2 141 Covance 6329-222 3M T-6295.6 PAGE: 2 APPENDIX 3 Individual Body Weight Data (kg) Covance 6329-222 3M T-6295.6 000278 142 Appendix 3 Individual Body Weight Data (kg) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T--6295) IN CYNOMOLGUS MONKEYS ANIMAL DAY DAY DAY NUMBER -14 -10 -7 GROUP: MALE 1 - 0 MG/KG/DAY 105349 105350 2.3 2.2 2.2 2.3 2.3 2.3 GROUP: MALE 2 - 0.02 MG/KG/DAY 105344 105347 105348 2.4 2.4 2.4 2.1 2.1 2.1 2.5 2.5 2.5 GROUP: MALE 3 - 2 MG/KG/DAY 105345 2.2 2.2 2.2 GROUP: FEMALE 1 - 0 MG/KG/DAY 105366 105367 2.0 2.0 2 .1 2.1 2.1 2.1 GROUP: FEMALE 2 - 0 . 0 2 MG/KG/DAY 105364 105365 10370 2.2 2.1 2.1 2.2 2.2 2.2 1.9 2.0 1.9 GROUP: FEMALE 3 - 2 MG/KG/DAY IK69 2.2 2.1 2.2 DAY -3 2.2 2.3 2.4 2.1 2.5 2.2 2.1 2.1 2.1 2.2 1.9 2.1 DAY -1 2.3 2.3 2.4 2.1 2.6 2.2 2.1 2.1 2.1 2.3 1.9 2.2 DAY i 2.2 2.4 2.4 2.1 2.5 2.2 2.1 2.1 2.2 2.3 2.0 2.2 DAY 5 2.2 2.3 2.4 2.1 2.4 2.2 2.0 2.0 2.1 2.3 2.0 2.1 DAY 8 2.4 2.5 2.5 2.3 2.5 2.3 2.2 2.1 2.2 2.4 2.0 2.2 Covance 6329-222 3M T-629S.6 PAGE : 1 DAY 12 2.4 2.5 2.4 2.2 2.4 2.2 2.1 2.1 2.2 2.3 2.0 2.2 DAY 15 2.3 2.5 2.4 2.3 2.4 2.3 2.1 2.2 2.2 2.4 2.0 2.1 143 000280 Appendix 3 Individual Body Weight Data (kg) 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS ANIMAL DAY DAY DAY NUMBER 19 22 26 GROUP: MALE. 1 - 0 MG/KG/DAY 105349 105350 2.4 2.4 2.4 2.5 2.5 2.5 GROUP: MALE 2 - 0.02 MG/KG/DAY 105344 105347 105348 2.4 2.5 2.4 2.3 2.3 2.3 2.4 2.4 2.5 GROUP: MALE 3 - 2 MG/KG/DAY 105345 2.3 2.1 2.1 GROUP: FEMALE 1 - 0 MG/KG/DAY 105366 105367 2.1 2.1 2.1 2.1 2.2 2.2 GROUP: FEMALE 2 - 0 . 0 2 MG/KG/DAY 105364 105365 105370 2.2 2.2 2.2 2.5 2.4 2.4 2.2 2.1 2.1 GROUP: FEMALE 3 - 2 MG/KG/DAY 105369 2.3 2.1 2.1 DAY 29 2.3 2.4 2.4 2.3 2.4 2.0 2.1 2.1 2.1 2.3 2.0 2.1 144 Covance 6329-222 3M T-6295.6 PAGE: 2 APPENDIX 4 Individual Clinical Hematology Data Individual Clinical Chemistry Data Covance 6329-222 3M T-6295.6 000251 ANIMAL NUMBER RBC XIOVuL Appendix 4 Individual Clinical Hematology Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS HGB HCT MCV MCH MCHC PLT RETIC RETIC G/DL % EL PG % XIOVfiL % XlO'/pL Group : 1 105349 105350 MEAN S .D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level: 0 5.41 5.88 13.0 15.0 5.64 .332 2 14.0 1.41 2 Dose Level: 0.02 4.97 5.49 5.09 12.3 12.7 12.9 5.18 .272 3 12.6 .31 3 Dose Level: 2.0 5.91 12.8 Dosage Unit: mg/kg/day 39.9 45.3 73.8 77.0 24.0 25.5 42.6 3.82 2 75.4 2.26 2 24.8 1.06 2 Dosage Unit: mg/kg/day 38.5 40.1 38.6 77.5 73.1 75.7 24.8 23.2 25.3 39.1 .90 3 75.4 2.21 3 24.4 1.10 3 Dosage Unit: mg/kg/day 41.0 69.4 21.7 32.6 33.2 32.9 .42 2 32.0 31.7 33.4 32.4 .91 3 31.2 404 600 502 138.6 2 .9 .9 .9 .00 2 473 358 531 454 88.1 3 1.3 .3 1.1 .9 .53 3 531 .7 49 53 51 2.8 2 65 16 56 46 26.1 3 41 Covance 6329-222 3M T-6295.6 000252 146 ANIMAL NUMBER WBC XloVpL Appendix 4 Individual Clinical Hematology Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS N-SEG LYMPH XlOVfiL XlOVpL MONO XlOVpL EOSIN XlOVpL BASO XIOVpL N -SEG% LYMPH% MONO% EOSIN% Covance 6329-222 3M T-6295.6 BASO% Group : 1 Dose Level : 0 Dosage Unit: mg/kg/day 105349 105350 9.1 4 .1 4.5 .4 .0 .0 45 49 5 0 0 7.7 2.6 4.6 .3 .2 .0 34 59 4 2 0 MEAN S .D. N 8.4 3.4 4.6 .4 .1 .0 40 54 4 1 0 .99 1.06 .07 .07 .14 .00 7.8 7.1 .7 1.4 .0 22 22 2 2 222 2 2 Group : 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105344 105347 105348 10.3 3.2 6.3 .6 .2 .0 31 61 6 2 0 8.8 1. 4 7.0 .4 .1 .0 15 79 4 1 0 8.4 4.0 4.2 .2 .0 .0 48 50 2 0 0 MEAN S. D. N 9.2 2.9 5.8 .4 .1 .0 31 63 4 1 0 1.00 1.33 1.46 .20 .10 .00 16.5 14.6 2.0 1.0 .0 33 3 3 3 3 3 33 3 3 Group: 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105345 8.2 2.8 4.8 .6 .0 .0 34 58 8 0 0 000283 147 Appendix 4 Individual Clinical Hematology Data Males Day -7 -WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 1 105349 105350 Group: 2 105344 105347 105348 Group: 3 105345 Dose Level: 0 Dose Level: 0.02 Dose Level: 2.0 Dosage Unit: mg/kg/day Dosage Unit: mg/kg/day Dosage Unit: mg/kg/day Covance 6329-222 3M T-6295.6 000284 148 ANIMAL NUMBER Appendix 4 Individual Clinical Hematology Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS RBC HGB HCT MCV MCH MCHC PLT RETIC XlOVpL G/DL % FL PG % XlOVjlL % Covance 6329-222 3M T-6295.6 RETIC XlOVfiL Group: 1 105366 105367 MEAN S.D. N Group: 2 105364 105365 105370 MEAN S.D. N Group: 3 105369 Dose Level: 0 5.02 5.48 12.4 14.3 5.25 .325 2 13.3 1.34 2 Dose Level: 0.02 4.77 4.98 5.25 13.0 12.7 12.6 5.00 .241 3 12.8 .21 3 Dose Level: 2.0 5.53 13.4 Dosage Unit: mg/kg/day 37.6 43.2 75.0 78.8 24.8 26.1 40.4 3.96 2 76.9 2.69 2 25.4 .92 2 Dosage Unit: mg/kg/day 38.9 39.0 39.3 81.5 78.3 74.9 27.2 25.5 24.1 39.1 .21 3 78.2 3.30 3 25.6 1.55 3 Dosage Unit: mg/kg/day 40.2 72.7 24.2 33.1 33.1 33.1 .00 2 33.3 32.6 32.2 32.7 .56 3 33.3 351 606 478 180.3 2 .7 1.0 .8 .21 2 355 365 570 430 121.3 3 .6 .9 .6 .7 .17 3 505 .8 35 55 45 14.1 2 29 45 32 35 8.5 3 44 0002S3 149 ANIMAL NUMBER WBC XlOVpL Appendix 4 Individual Clinical Hematology Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-629S) IN CYNOMOLGUS MONKEYS N-SEG XlOVpL LYMPH XIOVpL MONO XlOVpL EOSIN XIOVpL BASO N -SEG% XlOVfiL LYMPH% MONO% EOSIN% Covance 6329-222 3M T-6295.6 1BASO* Group : 1 Dose Level : 0 Dosage Unit: mg/kg/day 105366 7.9 2.2 5.2 .4 .1 .0 28 66 5 1 0 105367 10.0 4.9 5.0 .0 .0 .0 49 50 000 MEAN S. D. N 9.0 3.6 5.1 .2 .0 .0 38 58 2 0 0 1.48 1.91 .14 .28 .07 .00 14.8 11.3 3.5 .7 1 22 2 222 222 22 Group : 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105364 105365 105370 8.6 12.4 9.6 1.6 6.2 1.6 10.2 7.3 2.1 .5 .2 .5 .1 .3 .0 .0 19 73 .0 13 82 .0 75 21 620 410 300 MEAN S. D. N 10.2 3.5 6.2 .4 .1 .0 36 59 4 1 0 1.97 3.29 4.05 .12 .10 .00 34.2 32.9 1.5 1.0 33333333333 Group : 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105369 11.7 6.2 4.8 .4 .2 .0 53 42 410 000286 150 Appendix 4 Individual Clinical Hematology Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 1 Dose Level: 0 105366 - - - 105367 - - - Group: 2 Dose Level: 0.02 105364 - - - 105365 - - - 105370 - - - Group: 3 105369 Dose Level: 2.0 --- Dosage Unit: mg/kg/day --- Dosage Unit: mg/kg/day ---- Dosage Unit: mg/kg/day -- Covance 6329-222 3M T-6295.6 000287 151 ANIMAL NUMBER Appendix 4 Individual Clinical Hematology Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS RBC HGB HCT MCV MCH MCHC PLT RETIC XlOVnL G/DL % FL PG % XlOVnL % Covance 6329-222 3M T-6295.6 RETIC XlOVjlL Group : 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 6.00 6.05 13.6 14 .5 6.02 .035 2 14.0 .64 2 Dose Level: 0.02 4 .90 5.05 5.41 12.2 12.1 13.5 5.12 .262 3 12.6 .78 3 Dose Level : 2.0 5.69 13.2 Dosage Unit: mg/kg/day 42.7 45.3 71.0 74.8 22.6 24.0 44.0 1.84 2 72.9 2.69 2 23.3 .99 2 Dosage Unit: mg/kg/day 38.1 37.6 41.6 77.8 74.5 76.8 24.9 23.9 25.0 39.1 2 .18 3 76.4 1.69 3 24.6 .61 3 Dosage Unit: mg/kg/day 41.5 73.1 23.1 31.8 32.1 31.9 .21 2 32.0 32.1 32.5 32.2 .26 3 31.7 357 522 440 116.7 2 448 286 443 392 92,1 3 503 .9 .7 .8 .14 2 .5 .2 .9 .5 .35 3 .2 54 42 48 8. 2 24 10 49 28 19. 3 11 000288 152 ANIMAL NUMBER WBC XloVpL Appendix 4 Individual Clinical Hematology Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS N-SEG XlOVpL LYMPH XIOVpL MONO XloVpL EOSIN XloVpL BASO N-SEG* XlOVpL LYMPH* MONO% EOSIN% Covance 6329-222 3M T-6295.6 BASO* Group : 1 Dose Level 0 Dosage Unit: mg/kg/day 105349 105350 9.1 2.0 6.3 .7 .1 .0 22 69 8 1 0 9.4 1.3 7 .4 .4 .3 .0 14 79 4 3 0 MEAN S. D. N 9.2 1.6 6.8 .6 .2 .0 18 74 6 2 0 .21 .49 .78 .21 .14 .00 5.7 7.1 2.8 1.4 .0 222222 22222 Group : 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105344 105343 105348 10.9 .9 8.8 1.0 .2 .0 9 81 9 1 0 9.3 1.0 7.6 .4 .3 .0 10 82 5 3 0 6.1 1.2 4.4 .3 .1 .0 20 72 5 2 0 MEAN S.D. N 8.8 1.0 6.9 .6 .2 .0 13 78 6 2 0 2.44 .15 2.27 .38 .10 .00 6.1 5.5 2.3 1.0 .0 33333333333 Group : 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105345 5.8 .9 4 .4 .4 .0 .0 16 77 7 0 0 0002c CD 153 Appendix 4 Individual Clinical Hematology Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 1 Dose Level: 0 105349 - 105350 - - - Group: 2 Dose Level: 0.02 105344 - - - 105347 - - - 105348 - - - Group: 3 105345 Dose Level: 2.0 --- Dosage Unit: mg/kg/day -- Dosage Unit: mg/kg/day ---- Dosage Unit: mg/kg/day -- Covance 6329-222 3M T-6295.6 000290 154 ANIMAL NUMBER Appendix 4 Individual Clinical Hematology Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS RBC HGB HCT MCV MCH MCHC PLT RETIC XIOVpL G/DL % FL PG % XlOVnL % Covance 6329-222 3M T-6295.6 RETIC XloVpL Group: 1 105366 105367 MEAN S.D. N Group: 2 105364 105365 105370 MEAN S.D. N Group: 3 105369 Dose Level: 0 5.02 5.26 12.6 13.7 5 .14 .170 2 13.2 .78 2 Dose Level : 0.02 5.15 5.01 5.24 14 .1 12.7 12.5 5.13 .116 3 13.1 .87 3 Dose Level: 2.0 4.81 12.3 Dosage Unit: mg/kg/day 38.9 42.2 77.4 80.4 25.1 26.1 40.5 2.33 2 78.9 2.12 2 25.6 .71 2 Dosage Unit: mg/kg/day 43.1 40.5 39.8 83.7 80.7 75.9 27.4 25.4 23.8 41.1 1.74 3 80.1 3.93 3 25.5 1.80 3 Dosage Unit: mg/kg/day 36.7 76.4 25.6 32.5 32.5 32.5 .00 2 32.8 31.4 31.4 31.9 .81 3 33.6 382 640 511 182.4 2 1.0 1.3 1.2 .21 2 377 340 558 425 116.7 3 1.1 .7 .6 .8 .26 3 529 1.0 50 68 59 12.7 2 57 35 31 41 14.0 3 48 TS2000 155 ANIMAL NUMBER WBC XlOVnL Appendix 4 Individual Clinical Hematology Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS N-SEG XloVpL LYMPH XIOV^L MONO EOS IN XlOVjiL XlOVnL BASO N -SEG% XlOVpL LYMPH% MONO% EOSINI Covance 6329-222 3M T-6295.6 ]BASO! Group: 1 Dose Level : 0 Dosage Unit: mg/kg/day 105366 9.2 3.9 4 .4 .9 .0 .0 43 47 10 0 0 105367 13.0 3.7 8.0 1.2 .0 .0 28 62 900 MEAN S. D. N 11.1 3.8 6.2 1.0 .0 .0 36 54 10 0 0 2.69 .14 2.55 .21 .00 .00 10.6 10.6 .7 .0 22222 222222 Group : 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105364 8.7 1.7 6.2 .6 .1 .0 19 72 7 1 0 105365 12.9 2.2 10.1 .5 .1 .0 17 78 4 10 105370 8.3 2.6 5.0 .6 .1 .0 31 60 8 1 0 MEAN S .D. N 10.0 2.2 7.1 .6 .1 .0 22 70 6 1 0 2.55 .45 2.67 .06 .00 .00 7.6 9.2 2.1 .0 333 33 3 33 3 33 Group: 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105369 15.7 7.2 7.9 .5 .1 .0 46 50 3 1 0 000292 156 Appendix 4 Individual Clinical Hematology Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ANISO POLY POIK HYPO TOXNEUT Group: 1 105366 105367 Dose Level: 0 ----- Group: 2 105364 105365 105370 Dose Level: 0.02 Group: 3 105369 Dose Level: 2.0 --- Dosage Unit: mg/kg/day --Dosage Unit: mg/kg/day Dosage Unit: mg/kg/day -- Covance 6329-222 3M T-6295.6 000293 157 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group : 1 Dose Level : 0 Dosage Unit: mg/kg/day 105349 105350 95 25 1.0 8.2 5.2 3.0 .2 68 23 1.1 8.2 5.2 3.0 .3 8 111 9 199 46 81 MEAN S. D. N 82 19. 1 2 24 1 .4 2 1.0 .07 2 8.2 .00 2 5.2 .00 2 3.0 .00 2 .2 .07 2 8 155 .7 62.2 22 64 24.7 2 Group : 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105344 105347 105348 46 29 .9 7.7 4.6 3.1 .3 11 137 65 71 27 1.0 7.7 5.1 2.6 .4 9 202 82 66 31 1.0 7.8 4.6 3.2 .3 8 145 109 MEAN S. D. N 61 13.2 3 29 2.0 3 1.0 .06 3 7.7 .06 3 4 .8 .29 3 3.0 .32 3 .3 .06 3 9 161 1.5 35.4 33 85 22.2 3 Group : 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105345 54 28 1.0 8.2 5.0 3.2 .4 10 150 68 000294 158 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L ]P AMYL U/L Covance 6329-222 3M T-6295.6 Group: 1 105349 105350 MEAN S.D. N Group: 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 52 33 45 46 48 4.9 2 40 9.2 2 Dose Level : 0.02 58 45 43 71 48 59 50 7.6 3 58 13.0 3 Dose Level : 2.0 54 50 Dosage Unit: mg/kg/day 638 91 587 84 4 3 612 36.1 2 88 4.9 2 4 .7 2 Dosage Unit: mg/kg/day 606 78 767 143 670 81 7 5 5 681 81.1 3 101 36.7 3 6 1.2 3 Dosage Unit: mg/kg/day 1086 117 5 156 301 228 102.5 2 200 208 233 214 17.2 3 220 353 494 424 99.7 2 376 318 250 315 63.1 3 244 37 90 64 37.5 2 132 38 17 62 61.2 3 7 242 340 291 69 2 275 220 175 223 50 3 144 000295 159 Appendix 4 Individual Clinical Chemistry Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group : 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 10.6 10.8 7.6 6.8 10.7 .14 2 7.2 .57 2 Dose Level : 0.02 10.3 11.2 10.3 7.8 6.9 8.3 10.6 .52 3 7.7 .71 3 Dose Level : 2.0 11. 4 8.7 Dosage Unit: mg/kg/day 161 5.7 110 164 5.8 110 162 2.1 2 5.8 .07 2 110 .0 2 Dosage Unit: mg/kg/day 159 5.1 111 164 5.8 117 154 6.0 108 159 5.0 3 5.6 .47 3 112 4.6 3 Dosage Unit: mg/kg/day 162 5.9 111 Covance 6329-222 3M T-6295.6 000296 160 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group : 1 Dose Level : 0 Dosage Unit: mg/kg/day 105366 105367 SH 70 82 24 .9 7.6 4.8 2.8 24 1.1 8.3 5.4 2.9 .3 .7 8 131 0 153 66 86 MEAN S.D. N 76 8.5 2 24 .0 2 1.0 .14 2 8.0 .49 2 5.1 .42 2 2.8 .07 2 .5 .28 2 4 142 5.7 15.6 22 76 14.1 2 Group : 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105364 105365 105370 55 23 .9 8.2 5.2 3.0 .4 7 114 47 73 23 .8 7.9 4.8 3.1 .4 8 169 49 64 31 .9 7.6 4.7 2.9 .3 11 178 110 MEAN S.D. N 64 26 .9 7.9 4.9 3.0 9.0 4.6 .06 .30 .26 .10 3333 33 .4 .06 3 9 154 2.1 34.6 33 69 35.8 3 Group : 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105369 71 27 1.0 8.2 4.9 3.3 .3 12 141 67 000297 161 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Covance 6329-222 3M T-6295.6 Group : 1 105366 105367 SH MEAN S .D. N Group : 2 105364 105365 105370 MEAN S .D. N Group : 3 105369 Dose Level : 0 45 38 153 68 99 76.4 2 53 21..2 2 Dose Level : 0.02 60 45 64 73 51 55 58 6.7 3 58 14 .2 3 Dose Level : 2.0 53 55 Dosage Unit: mg/kg/day 546 81 3 640 78 10 593 66.5 2 80 2.1 2 6 4.9 2 Dosage Unit: mg/kg/day 755 62 849 80 881 79 5 4 3 828 65.5 3 74 10.1 3 4 1.0 3 Dosage Unit: mg/kg/day 683 110 3 290 3524 1907 2286.8 2 253 567 334 385 163.0 3 1094 356 312 334 31.1 2 281 369 321 324 44 .1 3 315 17 2 10 10.6 2 21 158 12 64 81.8 3 89 247 216 232 21 2 193 228 225 215 19 3 224 000298 162 Appendix 4 Individual Clinical Chemistry Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUCROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group : 1 105366 105367 SH MEAN S. D. N Group : 2 105364 105365 105370 MEAN S.D. N Group : 3 105369 Dose Level : 0 10.6 11.3 6.5 8.3 11.0 .49 2 7.4 1.27 2 Dose Level : 0.02 10.6 10.8 11.7 6.4 7.7 9.8 11.0 .59 3 8.0 1.72 3 Dose Level : 2.0 11.3 6.8 Dosage Unit: mg/kg/day 161 5.9 111 157 8.6 115 159 2.8 2 7.2 1.91 2 113 2.8 2 Dosage Unit: mg/kg/day 154 5.1 110 155 6.1 114 166 6.3 118 158 6.7 3 5.8 .64 3 114 4.0 3 Dosage Unit: mg/kg/day 165 6.0 115 Covance 6329-222 3M T-629S.6 000299 163 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group: 1 105349 105350 MEAN :.D. N Group: 2 105344 105347 105348 MEAN S. D. N Group: 3 105345 Dose Level : 0 52 29 62 21 57 25 7.1 5.7 22 Dose Level : 0.02 56 21 68 25 62 25 62 6.0 3 24 2 .3 3 Dose Level : 2.0 58 21 Dosage Unit: mg/kg/day .9 7.4 .9 7.3 5.0 4.9 .9 7.4 5.0 .00 .07 .07 222 Dosage Unit: mg/kg/day .8 7.1 .8 6.9 .9 7.1 4.4 4.6 4 .4 .8 7.0 4.5 .06 .12 .12 33 3 Dosage Unit: mg/kg/day .9 7.3 4.7 2.4 2.4 2.4 .00 2 2.7 2.3 2.7 2.6 .23 3 2.6 .4 .3 .4 .07 2 .3 .3 .4 .3 .06 3 .4 19 97 45 13 188 114 16 142 80 4.2 64.3 48.8 22 2 10 9 10 10 .6 3 149 184 157 163 18.3 3 60 56 90 69 18.6 3 9 151 50 000300 164 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Covance 6329-222 3M T-6295.6 Group : 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 65 41 85 71 75 14 .1 2 56 21.2 2 Dose Level ; 0.02 44 40 39 69 41 60 41 2.5 3 56 14.8 3 Dose Level : 2.0 54 55 Dosage Unit : mg/kg/day 692 88 761 79 2 3 726 48.8 2 84 6.4 2 2 .7 2 Dosage Unit: mg/kg/day 531 68 691 125 575 72 2 2 2 599 82.7 3 88 31.8 3 2 .0 3 Dosage Unit: mg/kg/day 1028 105 2 903 895 899 5.7 2 139 888 709 579 391.1 3 219 363 461 412 69.3 2 325 311 230 289 51.3 3 302 35 27 31 5.7 2 227 284 256 40.3 2 62 35 24 40 19.6 3 205 190 146 180 30.7 3 11 139 000301 165 Appendix 4 Individual Clinical Chemistry Data Males Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S .D. N Group: 3 105345 Dose Level : 0 10.1 10.3 7.6 6.1 10.2 .14 2 6.8 1.06 2 Dose Level : 0.02 9.7 7.0 9.6 6.6 9.2 6.4 9.5 .26 3 6.7 .31 3 Dose Level : 2.0 10.0 7.0 Dosage Unit: mg/kg/day 151 5.2 107 153 5. 1 106 152 1.4 2 5.2 .07 2 106 2 Dosage Unit: mg/kg/day 147 5.0 110 150 4.5 110 145 4.7 103 147 2.5 3 4.7 .25 3 108 4 3 Dosage Unit: mg/kg/day 152 5.0 104 Covance 6329-222 3M T-6295.6 ZOPOOO 166 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Females Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group: 1 Dose Level : 0 Dosage Unit: mg/kg/day 105366 105367 80 19 .8 7 .1 4.7 2.4 .4 60 19 .9 7.7 5.1 2.6 .5 8 143 9 147 34 50 MEAN S .D. N 70 19 .8 7.4 4.9 2.5 14 .1 .0 .07 .42 .28 .14 222222 .4 .07 2 8 145 .7 2.8 22 42 11.3 2 Group: 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105364 105365 105370 60 24 .9 7.5 5.0 2.5 .5 15 102 42 57 28 .8 7.3 4 .4 2.9 .5 16 140 59 53 21 .8 7.2 4.7 2.5 .4 17 157 107 MEAN S.D. N 57 24 .8 7.3 4 .7 2.6 .5 16 133 69 3.5 3.5 .06 .15 .30 .23 .06 1.0 28.2 33.7 3333333333 Group: 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105369 68 27 1.0 7.5 4.7 2.8 .4 17 136 47 COOOO 167 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Females Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Covance 6329-222 3M T-6295.6 Group: 1 105366 105367 MEAN S .D. N Group: 2 105364 105365 105370 MEAN S.D. N Group: 3 105369 Dose Level : 0 45 42 71 103 58 18 .4 2 72 43. 1 2 Dose Level : 0.02 54 43 59 76 50 53 54 4 .5 3 57 16. 9 3 Dose Level : 2.0 46 55 Dosage Unit: mg/kg/day 491 77 665 85 1 2 578 123.0 2 81 5.7 2 2 .7 2 Dosage Unit: mg/kg/day 730 60 833 74 851 74 2 3 2 805 65.3 3 69 8 .1 3 2 .6 3 Dosage Unit: mg/kg/day 672 100 2 534 512 523 15.6 2 281 543 455 426 133.3 3 308 404 600 502 138.6 2 298 376 380 351 46.2 3 291 43 35 39 5.7 2 256 393 324 96 2 21 33 39 31 9.2 3 185 204 234 208 24 3 84 184 000304 168 Appendix 4 Individual Clinical Chemistry Data Females Day 2 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group : 1 105366 105367 MEAN S. D. N Group : 2 105364 105365 105370 MEAN S. D. N Group : 3 105369 Dose Level : 0 9.7 10.7 6.5 6.9 10.2 .71 2 6.7 .28 2 Dose Level : 0.02 10.2 9.7 10.0 6.7 7.6 8.4 10.0 .25 3 7.6 .85 3 Dose Level : 2.0 10.0 6.5 Dosage Unit: mg/kg/day 150 5.0 108 156 5.9 108 153 4.2 2 5.4 .64 2 108 .0 2 Dosage Unit: mg/kg/day 146 5.1 104 149 4.7 104 158 4.8 108 151 6.2 3 4.9 .21 3 105 2.3 3 Dosage Unit: mg/kg/day 151 4.7 106 Covance 6329-222 3M T-6295.6 000305 169 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Males Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group : 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S.D. N Group: 3 105345 Dose Level : 0 84 20 78 20 81 4.2 2 20 .0 2 Dose Level : 0.02 67 20 71 21 65 23 68 3.1 3 21 1.5 3 Dose Level : 2.0 63 18 Dosage Unit: mg/kg/day .8 7.6 .9 8.0 4.7 5.0 .8 7.8 4.8 .07 .28 .21 222 Dosage Unit: mg/kg/day .8 7.5 4 .4 .7 7.3 4.5 .8 7.8 4.5 .8 7.5 4.5 .06 .25 .06 333 Dosage Unit: mg/kg/day .8 7.5 4.6 2.9 3.0 3.0 .07 2 3.1 2.8 3.3 3.1 .25 3 2.9 .3 7 92 47 .2 14 185 57 .2 10 138 52 .07 4.9 65.8 7.1 2222 .3 11 134 .3 9 171 .3 9 162 49 51 52 .3 10 156 51 .00 1.2 19.3 1.5 3333 .4 7 137 50 000306 170 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Males Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Covance 6329-222 3M T-6295.6 Group : 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level :: 0 62 37 40 56 51 15.6 2 46 13. 4 2 Dose Level : 0.02 42 40 41 66 35 66 39 3.8 3 57 15. 0 3 Dose Level : 2.0 46 49 Dosage Unit: mg/kg/day 714 80 825 80 0 0 770 78.5 2 80 .0 2 0 .0 2 Dosage Unit: mg/kg/day 477 65 677 125 514 65 0 0 0 556 106.4 3 85 34.6 3 0 .0 3 Dosage Unit: mg/kg/day 1012 98 0 305 193 249 79.2 2 110 875 212 399 415.4 3 270 349 445 397 67.9 2 361 330 290 327 35.6 3 260 28 38 33 7.1 2 238 306 272 48.1 2 42 35 16 31 13.5 3 250 222 202 225 24.1 3 0 139 000307 171 Appendix 4 Individual Clinical Chemistry Data Males Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 1 105349 105350 MEAN 3 .D . N Group: 2 105344 105347 105348 MEAN S.D. N Group: 3 105345 Dose Level: 0 10.0 10.5 7.3 6.6 10.2 .35 2 7.0 .49 2 Dose Level: 0.02 9.4 9.9 10.0 7.3 7.0 6.6 9.8 .32 3 7.0 .35 3 Dose Level: 2.0 10.3 6.6 Dosage Unit: mg/kg/day 153 5.0 112 158 5.3 112 156 3.5 2 5.2 .21 2 112 .0 2 Dosage Unit: mg/kg/day 151 4.5 112 156 5.0 114 149 5.0 109 152 3.6 3 4.8 .29 3 112 2.5 3 Dosage Unit: mg/kg/day 156 5.1 112 Covance 6329-222 3M T-6295.6 000308 172 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Females Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group: 1 Dose Level: 0 105366 105367 SO 25 93 16 MEAN S.D. N 86 20 9.2 6.4 22 Group: 2 Dose Level: 0.02 105364 105365 105370 74 18 68 21 77 19 MEAN S.D. N 73 19 4.6 1.5 33 Group: 3 Dose Level: 2.0 105369 73 18 Dosage Unit: mg/kg/day .8 8 .1 4.9 .8 8.0 4.2 .8 8.0 4.6 .00 .07 .49 222 Dosage Unit: mg/kg/day .8 8.1 .7 7.8 .9 8.1 5.1 4.5 4.9 .8 8.0 4.8 .10 .17 .31 333 Dosage Unit: mg/kg/day .8 7.9 4.6 3.2 3.8 3.5 .42 2 3.0 3.3 3.2 3.2 .15 3 3.3 3 6 128 42 2 8 131 23 2 7 130 32 07 1.4 2.1 13.4 2222 3 8 111 30 4 8 162 43 3 8 168 40 3 8 147 38 06 .0 31.3 6.8 3333 3 12 117 44 000309 173 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Females Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L 1P AMYL U/L Covance 6329-222 3M T-6295.6 Group: 1 105366 105367 MEAN S.D. N Group: 2 105364 105365 105370 MEAN S.D. N Group: 3 105369 Dose Level : 0 45 43 46 62 46 7 2 52 13.4 2 Dose Level: 0.02 58 65 45 53 43 53 49 8.1 3 57 6.9 3 Dose Level: 2.0 45 57 Dosage Unit: mg/kg/day 492 75 625 63 1 0 558 94.0 2 69 8.5 2 0 .7 2 Dosage Unit: mg/kg/day 655 57 805 72 865 75 0 1 0 775 108.2 3 68 9.6 3 0 .6 3 Dosage Unit : mg/kg/day 674 90 0 181 103 142 55.2 2 240 228 187 218 27.8 3 166 415 453 434 26.9 2 364 334 369 356 18.9 3 349 23 38 30 10.6 2 21 31 24 25 5.1 3 101 284 322 303 26 2 251 192 251 231 34 3 238 000310 174 Appendix 4 Individual Clinical Chemistry Data Females Day 7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 1 105366 105367 MEAN S. D. N Group: 2 105364 105365 105370 MEAN S. D. N Group: 3 105369 Dose Level : 0 10.4 11.5 6.5 5.7 11.0 .78 2 6.1 .57 2 Dose Level : 0.02 10.7 10.3 11.1 6.8 6.9 8.2 10.7 .40 3 7.3 .78 3 Dose Level : 2.0 10.6 6.4 Dosage Unit: mg/kg/day 155 5.7 112 158 7.0 113 156 2.1 2 6.4 .92 2 112 2 Dosage Unit: mg/kg/day 151 5.7 111 152 5.3 113 162 5.5 114 155 6.1 3 5.5 .20 3 113 1 3 Dosage Unit: mg/kg/day 155 5.3 113 Covance 6329-222 3M T-6295.6 000311 175 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Males Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group: 1 Dose Level : 0 Dosage Unit: mg/kg/day 105349 105350 89 18 70 20 .9 8.0 5.2 2.8 .8 7.4 4.8 2.6 .4 .4 6 100 9 182 33 43 MEAN S.D. N 80 19 .8 7.7 5.0 2.7 13.4 1.4 .07 .42 .28 .14 222222 .4 .00 2 8 141 2.1 58.0 22 38 7.1 2 Group : 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105344 105347 105348 60 18 .6 7.2 4.4 2.8 .4 10 136 37 61 20 .6 6.7 4.4 2.3 .4 7 158 55 67 19 1.0 7.6 4.7 2.9 .4 6 156 41 MEAN S.D. N 63 19 .7 7.2 4.5 2.7 3.8 1.0 .23 .45 .17 .32 333333 .4 .00 3 8 150 2.1 12.2 33 44 9.5 3 Group: 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105345 64 15 .7 7.4 4.8 2.6 .4 6 132 27 000312 176 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Males Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Covance 6329-222 3M T-6295.6 Group : 1 105349 105350 MEAN S .D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 52 34 42 47 47 7 .1 2 40 9.2 2 Dose Level : 0.02 42 41 34 48 31 57 36 5.7 3 49 8.0 3 Dose Level : 2.0 39 45 Dosage Unit: mg/kg/day 785 86 737 75 6 3 761 33.9 2 80 7.8 2 4 2.1 2 Dosage Unit: mg/kg/day 431 58 671 120 456 54 3 4 3 519 131.9 3 77 37.0 3 3 .6 3 Dosage Unit: mg/kg/day 908 89 2 131 193 162 43.8 2 97 85 151 111 35.2 3 245 372 415 394 30.4 2 351 294 272 306 40.8 3 242 41 29 35 8.5 2 248 277 2 62 20.5 2 52 37 15 35 18.6 3 236 188 184 203 28.9 3 2 123 000313 177 Appendix 4 Individual Clinical Chemistry Data Males Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group : 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 10.1 10.4 8.8 6.5 10.2 .21 2 7.6 1.63 2 Dose Level : 0.02 9.7 7.4 9.5 6.4 9.8 6.2 9.7 .15 3 6.7 .64 3 Dose Level : 2.0 10.1 6.4 Dosage Unit: mg/kg/day 152 4.6 105 150 5.0 106 151 1.4 2 4.8 .28 2 106 2 Dosage Unit: mg/kg/day 148 4.5 108 149 4.2 107 147 4.8 106 148 1.0 3 4.5 .30 3 107 1 3 Dosage Unit: mg/kg/day 153 4.8 108 Covance 6329-222 3M T-6295.6 000314 178 Covance 6329-222 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Females Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T -6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL TRIG MG/DL 3M T-6295.6 Group : 1 105366 105367 MEAN S.D. N Group : 2 105364 105365 105370 MEAN S.D. N Group : 3 105369 Dose Level : 0 77 22 82 16 80 3.5 2 19 4 .2 2 Dose Level : 0.02 49 20 57 22 67 17 58 9.0 3 20 2. 5 3 Dose Level : 2.0 59 18 Dosage Unit: mg/kg/day .7 7.4 .6 7.6 4.8 4.5 .6 7.5 4.6 .07 .14 .21 222 Dosage Unit: mg/kg/day .9 7.9 .5 7.4 .7 7.7 5.1 4.5 4.8 .7 7.7 4.8 .20 .25 .30 333 Dosage Unit: mg/kg/day .8 7.8 4.6 2.6 3.1 2.8 .35 2 2.8 2.9 2.9 2.9 .06 3 3.2 .4 .3 .4 .07 2 .5 .7 .4 .5 .15 3 .4 7 127 7 111 7 119 .0 11.3 22 6 111 8 150 9 155 8 139 1.5 24.1 33 8 107 27 20 24 4.9 2 28 42 36 35 7.0 3 41 occ r'j cn 179 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Females Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Covance 6329-222 3M T-6295.6 Group: 1 105366 105367 MEAN S.D. N Group : 2 105364 105365 105370 MEAN S.D. N Group : 3 105369 Dose Level :: 0 40 41 52 52 46 8.5 2 46 7 .8 2 Dose Level : 0.02 46 48 55 68 40 49 47 7.5 3 55 11. 3 3 Dose Level : 2.0 40 51 Dosage Unit: mg/kg/day 458 64 540 63 5 4 499 58.0 2 64 .7 2 4 .7 2 Dosage Unit: mg/kg/day 722 51 852 69 825 69 3 3 4 800 68.6 3 63 10.4 3 3 .6 3 Dosage Unit: mg/kg/day 727 86 3 157 361 259 144.2 2 341 383 197 307 97.5 3 462 399 456 428 40.3 2 289 335 337 320 27.2 3 440 18 41 30 16.3 2 31 67 20 39 24.6 3 309 262 313 288 36.1 2 190 189 222 200 18.8 3 283 000316 180 Appendix 4 Individual Clinical Chemistry Data Females Day 14 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group: 1 105366 105367 MEAN S. D. N Group: 2 105364 105365 105370 MEAN S. D. N Group: 3 105369 Dose Level : 0 10.1 io.e 6.6 7.0 10.4 .49 2 6.8 .28 2 Dose Level : 0.02 10.5 9.9 10.8 6.3 6.3 7.5 10.4 .46 3 6.7 .69 3 Dose Level : 2.0 10.4 6.1 Dosage Unit: mg/kg/day 152 5.5 110 157 6.5 109 154 3.,5 2 6.0 .71 2 110 .7 2 Dosage Unit: mg/kg/day 146 5.2 105 149 5.0 107 159 5.5 112 151 6. 8 3 5.2 .25 3 108 3.6 3 Dosage Unit: mg/kg/day 152 5.4 108 Covance 6329-222 3M T-6295.6 000317 181 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT <PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group: 1 Dose Level : 0 Dosage Unit: mg/kg/day 105349 105350 75 24 87 24 .8 7.5 4.6 2.9 .9 7.7 4.6 3.1 .3 .2 6 93 8 186 40 31 MEAN S.D. N 81 24 .8 7.6 4.6 3.0 8.5 .0 .07 .14 .00 .14 222222 .2 .07 2 7 140 1.4 65.8 22 36 6 2 Group: 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105344 105347 105348 66 22 .8 7.2 4.0 3.2 .2 9 129 48 73 22 .7 6.7 4.1 2.6 .2 7 151 55 80 29 1.0 8.7 4.8 3.9 .3 13 172 57 MEAN S.D. N 73 24 .8 7.5 4.3 3.2 .2 10 151 53 7.0 4.0 .15 1.04 .44 .65 .06 3.1 21.5 4 3 33 3 3 33 3 33 Group: 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105345 80 15 1.1 8.2 4.8 3.4 .3 8 91 25 000318 182 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Covance 6329-222 3M T-6295.6 Group : 1 103349 105350 MEAN S .D. N Group : 2 105344 105347 105348 MEAN S.D. N Group: 3 105345 Dose Level :; 0 61 36 43 56 52 12.7 2 46 14. 1 2 Dose Level : 0.02 42 45 34 49 49 93 42 7.5 3 62 26. 6 3 Dose Level : 2.0 31 38 Dosage Unit: mg/kg/day 798 103 712 90 6 2 755 60.8 2 96 9.2 2 4 2.8 2 Dosage Unit: mg/kg/day 477 73 710 143 505 65 2 4 2 564 127.2 3 94 42.9 3 3 1.2 3 Dosage Unit: mg/kg/day 815 88 1 476 344 410 93.3 2 134 202 606 314 255.2 3 202 362 485 424 87.0 2 382 350 270 334 57.7 3 285 37 37 37 .0 2 241 329 285 62.2 2 73 24 8 35 33.9 3 263 235 185 228 39.5 3 3 156 000319 183 Appendix 4 Individual Clinical Chemistry Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group : 1 105349 105350 MEAN S. D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 10.4 10.6 8.2 6.8 10.5 .14 2 7.5 .99 2 Dose Level : 0.02 10.0 10.0 10.5 6.7 5.9 6.3 10.2 .29 3 6.3 .40 3 Dose Level : 2.0 11.0 6.4 Dosage Unit: mg/kg/day 152 6.3 112 155 5.0 110 154 2.1 2 5.6 .92 2 111 1 2 Dosage Unit: mg/kg/day 149 4.8 108 152 4.5 112 155 6.0 110 152 3.0 3 5.1 .79 3 110 2 3 Dosage Unit: mg/kg/day 156 6.0 107 Covance 6329-222 3M T-6295.6 000320 184 ANIMAL NUMBER GLU MG/DL Appendix 4 Individual Clinical Chemistry Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS UN MG/DL CREAT MG/DL T PRO G/DL ALB G/DL GLOB G/DL T BILI MG/DL SBA umol/L CHOL MG/DL Covance 6329-222 3M T-6295.6 TRIG MG/DL Group: 1 Dose Level : 0 Dosage Unit: mg/kg/day 105366 105367 87 26 .8 7.4 4 .4 3.0 .2 11 107 41 86 24 .9 7.9 4 .3 3.6 .2 7 135 36 MEAN S .D. N 86 25 .8 7.6 4 .4 3.3 .7 1.4 .07 .35 .07 .42 2222 22 .2 .00 2 9 121 2.8 19.8 22 38 3 2 Group: 2 Dose Level : 0.02 Dosage Unit: mg/kg/day 105364 105365 105370 50 28 1.0 8.5 5.0 3.5 .4 8 114 45 71 22 .8 7.7 4 .4 3.3 .3 8 156 40 65 21 .8 7 .4 4.3 3.1 .2 11 158 52 MEAN S .D. N 62 10.8 3 24 3.8 3 .9 7.9 .12 .57 33 4.6 .38 3 3.3 .20 3 .3 .10 3 9 143 1.7 24.8 33 46 6 3 Group: 3 Dose Level : 2.0 Dosage Unit: mg/kg/day 105369 78 24 1.0 8.2 4.6 3.6 .2 17 62 27 000321 185 Covance 6329-222 ANIMAL NUMBER Appendix 4 Individual Clinical Chemistry Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS AST/SGOT ALT/SGPT ALK PHOS IU/L IU/L IU/L GGT IU/L SDH IU/L CK IU/L AMYLASE LIPASE IU/L IU/L P AMYL U/L Group: 1 105366 105367 MEAN S.D. N Group: 2 105364 105365 105370 MEAN S.D. N Group: 3 105369 Dose Level : 0 46 39 60 52 53 9.9 2 46 9..2 2 Dose Level : 0.02 53 49 44 60 39 52 45 7.1 3 54 5. 7 3 Dose Level : 2.0 36 47 Dosage Unit: mg/kg/day 418 70 516 75 9 4 467 69.3 2 72 3.5 2 6 3.5 2 Dosage Unit: mg/kg/day 926 67 966 79 832 81 4 2 2 908 68.8 3 76 7.6 3 3 1.2 3 Dosage Unit: mg/kg/day 637 93 2 335 262 298 51.6 2 215 363 240 273 79.2 3 341 444 633 538 133.6 2 290 348 381 340 46.1 3 326 18 241 130 157.7 2 10 39 20 23 14.7 3 18 300 440 370 99.0 2 193 205 257 218 34.0 3 221 3M -6295.6 000322 186 Appendix 4 Individual Clinical Chemistry Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER CA MG/DL I PHOS MG/DL NA MMOL/L K MMOL/L CL MMOL/L Group : 1 105366 105367 MEAN S. D. N Group : 2 105364 105365 105370 MEAN S. D. N Group : 3 105369 Dose Level : 0 10.3 10.6 5.6 7.1 10.4 .21 2 6.4 1.06 2 Dose Level : 0.02 11.2 10.4 11.2 7.9 6.6 6.2 10.9 .46 3 6.9 .89 3 Dose Level : 2.0 li.i 6.5 Dosage Unit: mg/kg/day 151 5.4 109 153 6.0 113 152 1.4 2 5.7 .42 2 111 2.8 2 Dosage Unit: mg/kg/day 152 5.5 109 153 5.4 112 160 5.6 113 155 4 .4 3 5.5 .10 3 111 2.1 3 Dosage Unit: mg/kg/day 158 5.7 110 Covance 6329-222 3M T-6295.6 00032a 187 Appendix 4 Individual Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUCROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER PCOAO IU/G Group: 1 105349 105350 MEAN S.D. N Group: 2 105344 105347 105348 MEAN S.D. N Group: 3 105345 Dose Level: 0 0 1 0 .7 2 Dose Level: 0.02 4 4 3 4 .6 3 Dose Level: 2.0 1 Dosage Unit: mg/kg/day Dosage Unit: mg/kg/day Dosage Unit: mg/kg/day Covance 6329-222 3M T-6295.6 000324 188 Appendix 4 Individual Clinical Chemistry Data Males Day 30 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER PCOAO IU/G Group : 1 105366 105367 MEAN S. D. N Group : 2 105364 105365 105370 MEAN S.D. N Group : 3 105369 Dose Level : 0 0 4 2 2.8 2 Dose Level : 0.02 7 0 3 3 3.5 3 Dose Level : 2.0 i Dosage Unit: mg/kg/day Dosage Unit : mg/kg/day Dosage Unit: mg/kg/day Covance 6329-222 3M T-6295.6 000325 189 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 000326 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 1 ANIMAL NUMBER: 105349 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2235.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 9:44 PROSECTOR: JILL PAUS RECORDER: WENDY CHERRY POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS MARROW, FEMUR (FM) : >SECTION EXAMINED; TISSUE NOT PRESENT TESTIS (TE) : -IMMATURE,-PRESENT THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY) , BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), TESTIS (TE), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR) , SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB) , PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY) , PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, MALE (MM), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP) , PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000327 191 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 2 ANIMAL NUMBER: 105350 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2295.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 11:00 PROSECTOR: JILL PAUS RECORDER: DONIA MEYER POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS MARROW, FEMUR (FM) : >SECTION EXAMINED; TISSUE NOT PRESENT TESTIS (TE) : -IMMATURE,-PRESENT THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), TESTIS (TE), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, MALE (MM), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000328 192 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 3 ANIMAL NUMBER: 105344 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2330.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 10:25 PROSECTOR: DAVID SCHUETTE RECORDER: NANCY DIEDRICH POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS MARROW, FEMUR (FM) : >SECTION EXAMINED; TISSUE NOT PRESENT TESTIS (TE) : -IMMATURE,-PRESENT THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), TESTIS (TE), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, MALE (MM), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) o oo CO N CD 193 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 4 ANIMAL NUMBER: 105347 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2125.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 11:00 PROSECTOR: DAVID SCHUETTE RECORDER: NANCY DIEDRICH POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS LIVER (LI) : -INFILTRATE, LYMPHOPLASMACYTIC, -MINIMAL TESTIS (TE) : -IMMATURE,-PRESENT THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY) , BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) , TESTIS (TE), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR) , ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, MALE (MM), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU) , SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000330 194 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 5 ANIMAL NUMBER: 105348 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2285.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 10:19 PROSECTOR: DENNIS HOFFMAN RECORDER: DONIA MEYER POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS MARROW, FEMUR (FM) : >SECTION EXAMINED; TISSUE NOT PRESENT KIDNEY (KD) : -INFILTRATE, LYMPHOPLASMACYTIC,-SLIGHT, TESTIS (TE) : -IMMATURE,-PRESENT FOCAL THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), TESTIS (TE), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR) , SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB) , PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, MALE (MM), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY) , BONE, FEMUR (FE), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000331 195 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 6 ANIMAL NUMBER: 105345 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 1870.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 11:10 PROSECTOR: WENDY CHERRY RECORDER: DENNIS HOFFMAN POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS MARROW, FEMUR (FM) : >SECTION EXAMINED; TISSUE NOT PRESENT TESTIS (TE) : -IMMATURE,-PRESENT THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP) , PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), TESTIS (TE), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, MALE (MM), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), PROSTATE (PR), SEMINAL VESICLES (SV), EPIDIDYMIDES (EP) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000332 196 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 7 ANIMAL NUMBER: 105366 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2000.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 9:40 PROSECTOR: DENNIS HOFFMAN RECORDER: DONIA MEYER POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS MARROW, FEMUR (FM) : >SECTION EXAMINED; TISSUE NOT PRESENT LIVER (LI) : -INFLAMMATION, LYMPHOHISTIOCYTIC,-SLIGHT, FOCAL THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, FEMALE (MF), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), OVARY (OV), UTERUS (UT), CERVIX (CV), VAGINA (VA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), KIDNEY (KD), LUNG (LU), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000333 197 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 8 ANIMAL NUMBER: 105367 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2085.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 9:47 PROSECTOR: DAVID SCHUETTE RECORDER: NANCY DIEDRICH POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS KIDNEY (KD) : -INFILTRATE, LYMPHOPLASMACYTIC,-MINIMAL LIVER (LI) : -INFILTRATE, LYMPHOPLASMACYTIC,-MINIMAL -LIPIDOSIS, SUBCAPSULAR (TENSION LIPIDOSIS),-SLIGHT, FOCAL THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, FEMALE (MF), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), OVARY (OV), UTERUS (UT), CERVIX (CV), VAGINA (VA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), LUNG (LU), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000334 198 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE : 9 ANIMAL NUMBER: 105364 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2080.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 8:09 PROSECTOR: DENNIS HOFFMAN RECORDER: DONIA MEYER POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS MARROW, FEMUR (FM) : >SECTION EXAMINED; TISSUE NOT PRESENT KIDNEY (KD) : -INFILTRATE, LYMPHOPLASMACYTIC,-SLIGHT ADRENAL, CORTEX (AC) : -HEMORRHAGE,-MODERATE, FOCAL -THROMBUS,-PRESENT >NOTE:UNILATERAL MICROSCOPIC FINDINGS THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, FEMALE (MF), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), OVARY (OV), UTERUS (UT), CERVIX (CV), VAGINA (VA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, MEDULLA (MA), THYMUS (TH) 000335 199 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 10 ANIMAL NUMBER: 105365 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 2245.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 8:08 PROSECTOR: DAVID SCHUETTE RECORDER: NANCY DIEDRICH POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -STAINS-PERINEUM/PERIANAL; DARK RED AND MOIST KIDNEY (KD) : -INFILTRATE, LYMPHOPLASMACYTIC,-MODERATE LIVER (LI) : -INFILTRATE, LYMPHOPLASMACYTIC,-MINIMAL THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, FEMALE (MF), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), OVARY (OV), UTERUS (UT), CERVIX (CV), VAGINA (VA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), LUNG (LU), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000336 200 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 11 ANIMAL NUMBER: 105370 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 1940.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 10:18 PROSECTOR: JILL PAUS RECORDER: WENDY CHERRY POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS LIVER (LI) : -INFILTRATE, LYMPHOPLASMACYTIC,-SLIGHT THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, FEMALE (MF), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), OVARY (OV), UTERUS (UT), CERVIX (CV), VAGINA (VA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000337 201 APPENDIX 5 Individual Animal Pathology Data Covance 6329-222 3M T-6295.6 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS;T-6295) IN CYNOMOLGUS MONKEYS PAGE: 12 ANIMAL NUMBER: 105369 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 05/22/98 STUDY DAY OF DEATH: 30 STUDY WEEK OF DEATH: 5 TERMINAL BODY WEIGHT: 1970.0 GRAMS DATE AND TIME OF NECROPSY: 05/22/98 8:08 PROSECTOR: JILL PAUS RECORDER: WENDY CHERRY POST-FIX WEIGHER: NOT AVAILABLE PATHOLOGIST: DR. TOM PALMER WEIGHER: NOT AVAILABLE NECROPSY OBSERVATIONS HISTOPATHOLOGY GENERAL COMMENT (GC) : -BONE MARROW SMEAR TAKEN -EYES - DAVIDSONS -NO MACROSCOPIC LESIONS THE FOLLOWING ORGANS WERE UNREMARKABLE AT NECROPSY: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH), MARROW, STERNUM (SE), BONE, STERNUM (SB), BRAIN (BR), SPINAL CORD (SC), NERVE, SCIATIC (SN), HEART (HT), MUSCLE, SKELETAL (SM), TRACHEA (TR), ESOPHAGUS (ES), GALLBLADDER (GB), PITUITARY (PI), AORTA (AO), LN, MESENTERIC (MS), THYROID (TY), PARATHYROID (PT), SALIV GL, MANDIB (SG), SKIN (SK), MAMMARY, FEMALE (MF), STOMACH, GL (ST), DUODENUM (DU), JEJUNUM (JE), ILEUM (IL), CECUM (CE), COLON (CO), RECTUM (RE), URINARY BLADDER (UB), OVARY (OV), UTERUS (UT), CERVIX (CV), VAGINA (VA) THE FOLLOWING TISSUES WERE UNREMARKABLE AT MICROSCOPIC EXAMINATION: EYE (EY), BONE, FEMUR (FE), MARROW, FEMUR (FM), KIDNEY (KD), LUNG (LU), LIVER (LI), SPLEEN (SP), PANCREAS (PA), ADRENAL, CORTEX (AC), ADRENAL, MEDULLA (MA), THYMUS (TH) 000338 202 APPENDIX 6 Covance 6329-222 3M T-6295.6 Quality Assurance Statement Summary and Individual Hormone Analyses Data Figure 1 - Mean Estradiol Data (pg/mL) - Males Figure 2 - Mean Estradiol Data (pg/mL) - Females Figure 3 - Mean Triiodothyronine Data (ng/dL) - Males Figure 4 - Mean Triiodothyronine Data (ng/dL) - Females NOTE: This appendix o f the report contains information supplied by Ani Lytics Inc. and has been reviewed by the Quality Assurance Unit o f Ani Lytics Inc. 000339 203 Covance 6329-222 ____ 3M T-6295.6 r# v f 1 1 INCORPORATED 200 Girard Street, Suite 200, Gaithersburg, MD 20877 301-921-0168 800-237-2815 The reports listed below was reviewed for compliance with the FDA Good Laboratory Practices and with the EPA Good Laboratory Practices. The final report and all associated raw data were reviewed for accuracy and consistency and the findincis were reported to management. The methods used were the methods described and the report accurately reflects the data. Therefore, these studies were done in compliance with the FDA and EPA Good Laboratory Practices. Franklin B. Newman QA Auditor SPONSOR: C 0 l//fy /C ^ /} 6 $ t M<L' STUDY: ,REPORT TYPE: Cl, 3 , T3 T*), AUDIT DATE: REPORT TO MGMT: AUDIT #: -fH-J ft-??/ 000340 204 Appendix 6 Summary and Individual Hormone Analyses Data Males Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ESTRADIOL ESTRONE ESTRIOL THYROID STIMULATING TRIIODOTHYRONINE THYROXIN pg/mL pg/mL ng/mL (JlU/mL ng/dL pg/dL Group : 1 105349 105350 MEAN S. D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 34.86 29.20 38.62 17.83 32.03 4.002 2 28.23 14.701 2 Dose Level : 0.02 24.95 34 .14 26.73 18.44 17.74 15.79 28.61 4 .874 3 17.32 1.373 3 Dose Level : 2.0 28.71 13.98 Dosage Unit: mg/kg/day 0.00 0.00 2.33 3.30 0.00 0.000 2 2.82 0.686 2 Dosage Unit: mg/kg/day 0.00 0.00 0.00 2.62 2.49 1.21 0.00 0.000 3 2.11 0.779 3 Dosage Unit: mg/kg/day 0.00 1.71 160.41 119.07 139.74 29.232 2 152.65 148.40 66.41 122.49 48.610 3 97.61 4.01 6.84 5.43 2.001 2 3.90 4.43 4.52 4.28 0.335 3 3.86 Covance 6329-222 3M T-6295.6 000341 205 Appendix 6 Summary and Individual Hormone Analyses Data Females Day -7 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ESTRADIOL ESTRONE ESTRIOL THYROID STIMULATING TRIIODOTHYRONINE THYROXIN pg/mL pg/mL ng/mL |iIU/mL ng/dL (ig/dL Group: 1 105366 105367 MEAN S.D. N Group : 2 105364 105365 105370 MEAN S.D. N Group: 3 105369 Dose Level : 0 45.43 45.13 25.12 32.42 45.28 0.212 2 28.77 5.162 2 Dose Level : 0.02 28.17 70.25 45.95 21.98 26.58 42.65 48.12 21.124 3 30.40 10.852 3 Dose Level : 2.0 37.37 29.50 Dosage Unit: mg/kg/day 0.00 0.00 1.26 7.48 0.00 0.000 2 4.37 4.398 2 Dosage Unit: mg/kg/day 0.00 0.00 0.00 3.07 3.65 0.80 0.00 0.000 3 2.51 1.506 3 Dosage Unit: mg/kg/day 0.00 1.41 172.31 92.99 132.65 56.088 2 148.92 176.45 67.89 131.09 56.434 3 122.38 5.55 4.21 4.88 0.948 2 5.71 4.94 3.62 4.76 1.057 3 3.60 Covance 6329-222 3M T-6295.6 000342 206 Appendix 6 Summary and Individual Hormone Analyses Data Males Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ESTRADIOL ESTRONE ESTRIOL THYROID STIMULATING TRIIODOTHYRONINE THYROXIN pg/mL pg/mL ng/mL pIU/mL ng/dL pg/dL Group : 1 105349 105350 MEAN S.D. N Group : 2 105344 105347 105348 MEAN S.D. N Group : 3 105345 Dose Level : 0 33.04 32.75 25.02 18.84 32.90 0.205 2 21.93 4.370 2 Dose Level : 0.02 25.19 37.25 40.32 10.24 14.20 19.73 34.25 7.998 3 14.72 4.767 3 Dose Level : 2.0 18.16 20.12 Dosage Unit: mg/kg/day 0.00 0.00 0.73 3.57 0.00 0.000 2 2.15 2.008 2 Dosage Unit: mg/kg/day 0.00 0.00 0.00 3.27 3.58 2.45 0.00 0.000 3 3.10 0.584 3 Dosage Unit: mg/kg/day 0.00 0.95 181.44 191.25 186.35 6.937 2 195.89 209.96 159.16 188.34 26.229 3 90.73 3.32 5.31 4.32 1.407 2 3.86 3.82 4.90 4.19 0.612 3 3.18 Covance 6329-222 3M T-6295.6 000343 207 Appendix 6 Summary and Individual Hormone Analyses Data Females Day 29 4-WEEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS ANIMAL NUMBER ESTRADIOL ESTRONE ESTRIOL THYROID STIMULATING TRIIODOTHYRONINE THYROXIN pg/mL pg/mL ng/mL pIU/mL ng/dL Hg/dL Group : 1 105366 105367 MEAN S.D. N Group : 2 105364 105365 105370 MEAN S.D. N Group : 3 105369 Dose Level : 0 67.67 39.27 24.46 27.47 53.47 20.082 2 25.97 2.128 2 Dose Level : 0.02 31.90 87.09 57.17 32.66 30.94 31.23 58.72 27.628 3 31.61 0.921 3 Dose Level : 2.0 19.82 25.85 Dosage Unit: mg/kg/day 0.00 0.00 2.57 3.32 0.00 0.000 2 2.95 0.530 2 Dosage Unit: mg/kg/day 0.00 0.00 0.00 2.50 4.64 1.14 0.00 0.000 3 2.76 1.764 3 Dosage Unit: mg/kg/day 0.00 3.92 141.93 183.46 162.70 29.366 2 179.61 196.25 165.89 180.58 15.203 3 90.17 4.57 3.95 4.26 0.438 2 5.26 3.79 3.09 4.05 1.108 3 3.38 Covance 6329-222 3M T-629S.6 000344 208 000345 40 38 36 34 ~ 32 -> 30 28 E 26 -3 QOl)) 24 J 22 20 18 -3 16 14 12 -3 10 -7 Figure 1 Mean Estradiol Data (pg/mL) -Males Covance 6329-222 3M T-6295.6 --O 0 mg PFOS/kg/day 0.02 mg PFOS/kg/day 2.0 mg PFOS/kg/day Days 209 '-A 29 Figure 2 Mean Estradiol Data (pg/mL) - Females Covance 6329-222 3M T-6295.6 000346 210 Figure 3 Mean Triiodothyronine Data (ng/dL) - Males Covance 6329-222 3M T-6295.6 000347 211 Figure 4 Mean Triiodothyronine Data (ng/dL) - Females Covance 6329-222 3M T-6295.6 000348 212 APPENDIX 7 Dose Confirmation Analysis Report Compound Stability Report Certificate of Analysis Quality of Assurance Statement Covance 6329-222 3M T-6295.6 NOTE: This appendix o f the report contains information supplied by the Sponsor and has been reviewed by the Quality Assurance Unit o f 3M. 000349 213 Covance 6329-222 3M T-6295.6 Dose Confirmation Analysis Report for Study title: 4-Week Capsule Toxicity Study with Perfluorooctanesulfonic Acid Potassium Salt (PFOS) in Cynomolgus Monkeys Covance 6329-222, (3M Medical Dept number: T- 6295.6). Author: Andrew M. Seacat Final Report Date: 2/28/01 Introduction Dose confirmation analysis were performed on samples of perfluorooctanesulfonic acid potassium salt (KPFOS) triturated in lactose. The 2.0 mg/kg/day dose level was prepared at 1:9 w:w PFOS: lactose, and the 0.02 mg/kg/day dose level was prepared at 1:999 w:w PFOS:lactose (sample numbers: CL1 # 1205A1, and CLI# 1206A1, respectively). These samples were prepared at Covance on 4-13-98 at the initiation of the in-life phase of the study. Eight samples were collected. Samples from the top, middle and bottom of each mixture were collected on 4-13-98 and sent to 3M Environmental Lab. Additional samples were collected to assess compound stability in the dosing vehicle from the middle of each mixture on 5-22-98, and were sent to the 3M Environmental Lab for analysis. Method summary The dose confirmation data were collected according to a method described fully in the Analytical Laboratory Report (1). Briefly, dose confirmation was performed by diluting the lactose dose samples 1000-fold with Milli-Q water, which were then extracted using the ion-pair reagent tetrabutylammonium hydrogen sulfate according to the procedure (1). The extracts from CLI # 1205A1 and CLI# 1206A1 were then diluted 1:200 and 1:2, respectively, to bring the PFOS levels in the linear range of the instrument. For each sample, (top, middle and bottom), the average PFOS spiked matrix extract value was used as correction factor for calculating the percent recovery. In all cases, samples were analyzed versus an unextracted curve using HPLC-ESMS/MS. Results The results indicated that the average SD for the 2.0 mg/kg/day dose level prepared at a 1:9 dilution in lactose was 80 0.02% of the target concentration. The average SD for the 0.02 mg/kg/day dose level prepared at a 1:999 dilution in lactose was 54 0.02% of the target concentration (See Table 1 and calculations in the appendix). The stability samples that were obtained from the middle of each mixture on 5-22-98 were 79% and 71% of the target concentration for the 2.0 and the 0.02 mg/kg/day dose level preparations respectively. Thus the dose preparations were stable for the entire dosing period. S ig n a tu re : / 7V Andrew M. Seacat Ph.D. 000350 214 Covance: 6329-222 3M T-6295.6 Dose Analysis Report Covance 6329-222 3M T-6295.6 Reference: 1. Hansen K. J. Analytical Laboratory Report from the 26-Week Capsule Toxicity Study with Perfluorooctanesulfonic Acid Potassium Salt (PFOS) in Cynomolgus Monkeys on the Determination of the Presence and Concentration of Perfluorooctanesulfonate (PFOS) in Liver and Serum Samples. Project Identification: Covance 6329-222, 6329-223, 3M Medical Dept number: T- 6295.7, Analytical study: FACT TOX030, 3M Laboratory request No. U2279. 214 pp, 2000. 000351 215 000352 Covance: 6329-222 3M T-6295.6 Table 1 Dose Analysis Report Appendix Study Produci Number(Tcst Substance) Matrix Method/Revtsion Analytical Lquipment System Number Instrument Software/Version Date o f lixtractmn/Analyst Date o f Analysis/Analyst Date o f Data Rcduction/Analyst 0Covance 6329-222. Dosing Vehicle FACT-M-3.0 and ETS-8-5.1- unextracted curves Soup 020199 Masslynx 3 4 10/23/00 RWW 10/24/00 KJH 10/25/00 KJH Filename: See Attachments R-Squared Value: See Attachments Slope: See Attachments Y-Intercept See Attachments Lactose Dosing Confirm ation Croup Dose Sample H Eipected Cone. PFOS ng/mL PFOS-Bckgrad Cooc. ng/mL PFOS % Recovery Accuracy Av, SD Method Hlk Lactl02300-WBIk-| 000 Lactl02300-WBlk-2 0.00 QC CLII205AI(l:9)-M S-l CUI205A1{I:9)-MSD-I CLI1206AKI 999)-MS-l CLD206A 1(1 999)-MSD-l 252 252 251 251 Dose CLI1205A 1(1:9) CL 11206AI ( 1 999) 535 544 CLU206AI G2-Top (1.999) $10 CLI 1206A1 02-Middle ( 1 999) 502 CLM206AI G2-Bottom{ 1:999) 509 Cl.11205A 1 G3-Top(l 9) CL11205A1 G3-Middle (1.9) CL11205AI G3-Botiom (1.9) 51! 507 513 Limit o f Quantitation Limit (LOQ): PFOS * 5.01 ng/mL on 9/19/00 and 25 I ng/mL on 9/26/00 PFOS " Periluorooctancsulfonaie NA NA NA NA 216 86% 200 80% 265 106% 276 110% NA 79% NA 71% NA 54% NA 52% NA 57% NA 81% NA 81% NA 71% NR - Sample not received nor reported NA - Not Applicable 54% 80% Date Untercd/Analyst Date Veriliod/Anaiyst 11/02/00 LAC kjh 12/22/00 2% 2% 216 Covance 6329-222 3M T-6295.6 3 Covance 6329-222 3M T-6295.6 Covance: 6329-222 3M T-6295.6 Dose Analysis Report Example Calculations: Sample 1:9 G3 l op Sample 1 999 G2 Top Matrix Spike 1.9 Matrix Spike 1 999 1 mg PFOS 10 mg lactose mix I me PFOS 1000 mg lactose mix 1006 ug mL 50.1 m: mL 102.1 me lactose mix 100 mL H20 101,9 me Inclose mix 100 mL H20 x 0.05 mL 1 mL x 0.01 ml, 1 mL -0.1021 m tPfO S mL H20 mL H 20 Dilution x1 200 Dilution xl 2 x 1000 ug 1 mg x 1000 ue 1 mg 0.252 ug mL 0.251 ug mL Dilution x1 200 Dilution x1 2 Matrix Spike Curve and Spike arc absolute and do not need purity/sait correction (PI'OS spkd sample] ng/mL x extraction dilution factor [PFOS Corrected spkd sample) (ng/mL) (PI US Corrected spkd sample) (ng/niL) [PFOS Corrected sample) ng/m l IPFOS sD tk e l measured [Expected PPOS] Sample [Expected PFOS] (ng/mL) x 100% x Sail and Purity Correction (PFOS spike) measured (ng/mL) "=H% Recovery * (Corrected, expected PFOS) ng/mL Oil! ug mL 0.510 uv mL [Measured PFOS) ng/m l x Purity Correction Factor x Extraction Dil Factor m[Concsted, measured PFOS) ICorresn-.it measmcd PF0.S1 ng/ml. X 100% (Corrected, expected I'KIS] ng/mL - % Recovery 4 217 000353 Covancc: 6329-222 3M T-6295.6 Dose Analysis Report CL205A-MS-1 486 ng/mL X 1.25 = 608 r.g/mL 608 ng/mL- (314 ng/mL x 1.25) =215.5ng/mL 215.5 ne/mL 252 nR/mL X 100 = 86% G2 Top 510 ng/mL X 0.9275 x0.8690 =411 ng/mL 203 ng/mL X .8690 X 1.25 = 220 ng/mL 220 ng/mL 411 ng/mL X 100% = 54% Covance 6329-222 3M T-6295.6 5 218 3M Corporate Health Phyaiee Corporate Occupational Medicine Corporate Product Responeibllity Corporate Toiicology 3M Medical Department September 6 ,2000 3M Center, 220-2E-02 PO Bo* 33220 St. Paul, MN 55133-3220 631 733 1110 Covance 6329-222 3M T-6295.6 Peter J. Thomford, Ph.D. Study Director, Toxicology Covance Laboratories Inc. 3301 Kinsman Blvd. Madison W I53704 Re. Covance 6329-222 (T-6295.6), 6329-223 (T-6295.7). Perfluorooctanesulfonate Compound Stability Report Peter: The perfluorooctanesulfonic acid potassium salt (PFOS, FC-95 Lot 217) used in Covance 6329-222, 6329-223, has remained stable for the entire duration o f the study. A Certificate o f Analysis (C o f A) dated March 9,2000 (1), identified the compound as being 90.49% C8F17S03-K+ by a combination ofLC/M S, 'H-NMR, l9F-NMR, and elemental analysis techniques. Previously, on December 1!11997, 19F-NMR was used to characterize FC-95 lot 217 and showed that the isomer distribution was identical to the C o f A sent to you earlier this year (2). Additionally, ,9F-N M R and 1H NM R conducted on August 24th 2000 (3) were compared to the l9F-N M R completed on December 1st 1997 and indicated that there was no significant differences in the composition o f this lot o f PFOS over intervening 3 and a h alf year time period. Thus, PFOS, FC-95 Lot 217 used was stable for greater the tim e period during which these studies were conducted. Sincerely, Andrew M. Seacat Ph.D. Toxicology Specialist 000355 219 Peter Thomford, Ph.D. Page 2 September 6,2000 Covance 6329-222 3M T-6295.6 References: 1. PayferR.M. Certificate O f Analysis FC-95, Lot 217. 3M Specialty Chemicals Division. March 9,2000 2. Kestner T. Fluorochemical Isomer Distribution by I9F-NMR Spectroscopy. FC-95, lot 217 Analytical Request 53030 . SA&C Analytical Lab. December 1,1997 3. Kestner T. Chemical Characterization of PFOS (FC-95, lot 217) by `H-NMR & 19F-NMR Spectroscopy. Comparison to FC-95/217 analysis from Req.#53030Request No. 61886. 3M SA&C Analytical Lab - 236-2B-11. August 24,2000. 000356 Covance 6329-222 3M T-6295.6 Centre Analytical Laboratories, Inc, 3048 Research Drive Phone: (814) 231-8032 State College, PA 16801 www.centrelab.com Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Revision 3 C entre Analytical Laboratories COA Reference #: 023-018A 3M Product: PFO S, Lot 217 R eference#: SD-018 ______________________P a rity : 86.9% ________ Test Name Specifications P u rity 1 Result 86.9% Appearance Identification NMR Metals (ICP/MS) 1. Calcium 2. Magnesium 3. Sodium 4. Potassium2 5. Nickel 6 . Iron 7. Manganese Total % Impurity (NMR) Total % Impurity (LC/MS) Total % Impurity (GC/MS) Related Compounds - POAA Residual Solvents (TGA) Purity by DSC Inorganic Anions (IC) 1. Chloride 2. Fluoride 3. Bromide 4. Nitrate 5. Nitrite 6 . Phosphate 7. Sulfate4 Organic A cid s5 (IC) 1. TFA 2. PFPA 3. HFBA 4. NFPA Elemental Analysis": 1. Carbon 2. Hydrogen 3. Nitrogen 4. Sulfur 5. Fluorine White Crystalline Powder 1. Theoretical Value = 17.8% 2. Theoretical Value = 0% 3. Theoretical Value = 0% 4. Theoretical Value = 5.95% 5. Theoretical Value = 60% Conforms Positive 1. 0.005 wt./wt.% 2 . 0 . 0 0 1 wt./wt.% 3. 1.439 wt./wt.% 4. 6.849 wt./wt.% 5. <0 . 0 0 1 wt./wt.% 6 . 0.005 wt.Avt.% 7. <0.001 wt./wt.% 1.91 wt./wt.% 8.41 wt./wt.% None Detected 0.33 wt./wt.% None Detected Not Applicable'' 1. <0.015 wt./wt.% 2. 0.59 wt./wt.% 3. <0.040 wt./wt.% 4. <0.009 wt./wt.% 5. <0.006 wt./wt.% 6 . <0.007 wt./wt.% 7. 8.76 wt./wt.% 1 . <0 .1 wt./wt.% 2 . <0 .1 wt.Avt.% 3. 0.10 wt.Avt.% 4. 0.28 wt./wt.% 1. 12.48 wt.Avt.% 2. 0.244 wt./wt.% 3. 1.74 wt.Avt.% 4. 8.84 wt./wt.% 5. 54.1 wt./wt.% COA023-018A Page 1 of 3 000357 Covance 6329-222 ____ 3M T-6295.6 Centre Analytical Laboratories, Inc. 3046 Research Drive Phone: (814) 231-8032 State College, PA 16801 www.centrelab.com Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Revision 3 C entre Analytical L aboratories COA Reference #: 023-018A D ate o f L ast Analysis: 08/31/00 Expiration D ate: 08/31/06 Storage Conditions: Frozen <-10C Re-assessm ent Date: 08/31/06 'P urity = 100% - (sum o f metal im purities, 1.45% +LC/M S im purities, 8.41% +Inorganic Fluoride, 0.59% +N M R im purities, 1,905% +organic acid im purities, 0.38% +PO A A , 0.33% ) Total im purity from all tests = 13.07% Purity = 100% - 13.07% = 86.9% 2P o tassiu m is expected in this salt form and is therefore not consid ered an im purity. 3P u rity b y D S C is generally not applicable to m aterials o f low purity. N o end o th erm w as observed for this sample. 4S u lfu r in th e sam p le appears to b e converted to S 0 4 and hence detected u sin g the inorganic anion method conditions. The anion result agrees well with the sulfur determ ination in the elem ental analysis, lending confidence to this interpretation. Based on the results, the SO4 is not considered an impurity. ST F A HFBA NFPA PFPA Trifluoroacetic acid H eptafluorobutyric acid Nonafluoropentanoic acid Pentafluoropropanoic acid th e o r e t i c a l value calculations based on th e em pirical form ula, C 8F 17SO 3T C (M W =538) This w ork w as conducted under EPA G ood Laboratory Practice Standards (40 C FR 160). COA023-018A Tage 2 o f 3 000358 222 Covance 6329-222 3M T-6295.6 Centre Analytical Laboratories, Inc. 3048 Research Drive Slate College, PA 16801 www.centrelab.com Phone: (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 INTERIM CERTIFICATE OF ANALYSIS Revision 3 C entre A nalytical L aboratories CO A Reference #: 023-018A LC/M S Purity Profile: Im purity C4 C5 C6 C7 T otal w L /w t % 1.22 1.33 4.72 1.14 8.41 Note: The C4 and C6 values were calculated using the C4 and C6 standard calibration curves, respectively. T he C5 value was calculated using the average result from the C4 and C6 standard curves. Likewise, the C7 value w as calculated using the average result from the C6 and C8 standard curves. P re p ared B y: ____a ___ t & O Charles Simdfrs Scientist, Centre Analytical Laboratories /o /n /o ! D ate iL ii fLbctReviewed By: 5hn Flaherty / Date Laboratory Manager, Centre Analytical Laboratories COA023-018A Page 3 of 3 000359 223 3M Environm ental Laboratory Note to File Covance 6329-222 3M T-6295.6 FormETS-4-15.0 - 224 ExactCopy of Original _ Io I r / dI Initial Date 000360 00361 Covance 6329-222 3M T-6295.6 Sponsor Protocol No.: FACT-TCR001 Centre Study No.: 023-018 QUALITY ASSURANCE STATEMENT Centre Study Number 023-018, entitled, "Characterization Study o f PFOS, Lot 217 and Lot 171" was reviewed by Centre Analytical Laboratories' Quality Assurance Unit. A ll reviewed phases were reviewed for conduct according to Centre Analytical Laboratories' Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings w ere reported to the Study Director and to management. Phase 1. Protocol Review 2. Label Checks (solubility) 3. Standard Solution Preparation and analysis by instrum entation 4. Raw Data Review (NMR) 5. Standard Solution Preparation (Solubility) 6. Standard Solution Preparation 7. Report Review (DSC/TGA) Date Inspected 5/19/00 7/14/00 6/20,21/00 6/22/00 7/11-13/00 7/14/00 6/29, 30/00 7/26/00 8/8/00 Date Reported to Study Director and Centre Management Date Reported to Sponsor Management 5/19/00 7/14/00 7/17/00 Pending 6/22/00 6/22/00 7/14/00 7/14/00 7/15/00 7/20/00 Pending Pending 6/30/00 7/27/00 8/8/00 7/14/00 Pending Pending 8. D ata Review and Interim Report Review 8/8,9/00 8/9/00 Pending M,-i/ n' r L isa Ivianning Quality A ssurance Officer Centre Analytical Laboratories, Inc. P-7-O D ate it'. \tl tSXACT COPY O f fH ORIGINAL DOCUMENT" c_s> DATE Page 3 o f 59 0003b2 226 APPENDIX 8 Cell Proliferation Report Covance 6329-222 3M T-6295.6 NOTE: This appendix o f the report contains information supplied by Pathology Associates International and has been reviewed by the Quality Assurance Unit of Pathology Associates International. 000363 Covance 6329-222 ____ 3M T-6295.6 Pathology Associates International A Com pany o f Science Applications International Corporation An& ipkpM O anatfCeow anx FINAL CELL PROLIFERATION REPORT 4-WEEK CAPSULE TOXICITY STUDY W ITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS COVANCE STUDY NUMBER 6329-222 PREPARED FOR: 3M TOXICOLOGY SERVICES BUILDING 220-2E-02,3M CENTER ST. PAUL, MN 55144-1000 PREPARED BY: PATHOLOGY ASSOCIATES INTERNATIONAL 15 WORMAN'S MILL COURT, SUITE I FREDERICK, MD 21701 NOVEMBER 11,1999 15 W orman's Mill Court, Suite I * Frederick, M aryland 21701 (301) 663-1644 * (301) 663-8994 FAX 000364 228 Covance 6329-222 3M T-6295.6 Final Cell Proliferation Report Covance Study Number 6329-222 Page 2 CELL PROLIFERATION REPORT 4-W EEK CAPSULE TOXICITY STUDY WITH PERFLUOROOCTANE SULFONIC ACID POTASSIUM SALT (PFOS; T-6295) IN CYNOMOLGUS MONKEYS COVANCE STUDY NUMBER 6329-222 PURPOSE The purpose of the study was to provide data for determining an estimated maximum-tolerated dose and lower dose levels to be used in a chronic toxicity study and to assess the effect of the test material on critical enzyme levels, hormones, and other selected biochemical parameters. This report, submitted by Pathology Associates International (PAI) to the study Sponsor, 3M, represents the cell proliferation findings and interpretation for Covance Study Num ber 6329-222 entitled "4-W eek Capsule Toxicity Study with Perfluorooctane Sulfonic A cid Potassium Salt (PFOS; T-6295) in Cynomolgus M onkeys". All aspects o f the tasks associated with PAI's portion of this study were conducted in compliance with the Environmental Protection Agency Good Laboratory Practice (GLP) Regulations as set forth in Title 40 of the US Code of Federal Regulations, Part 792, issued November 29, 1983 (effective December 29, 1983), and with any applicable amendments. MATERIALS AND METHODS Tissue Collection for Cell Proliferation Two animals per sex in control group 1, three animals per sex in dose group 2 (0.02 mg/kg/day) and one animal per sex in dose group 3 (2.0 mg/kg/day) were sacrificed after four weeks on diet A representative sample of the liver, testes (males only) and pancreas from each animal was fixed in zinc formalin, processed and embedded to paraffin block by Covance per protocol specifications. Tissue blocks were then shipped to PAI for sectioning and staining. From each block, a slide was prepared for H& E evaluation and immunohistochemical detection of proliferating cell nuclear antigen (PCNA), a marker of cell proliferation. Im munohistochemistry for Cell Proliferation Sections of paraffin-embedded tissues were cut at 5 pm and placed on positively charged slides (Superfrost Plus, Fisher Scientific, Pittsburgh, PA) to ensure adhesion during processing for PCNA. Standard im m unohistochemical methods were used to stain tissues for PCNA (PAI's Standard Operating Procedure for Immunohistochemical Staining (SOP #707). Briefly, tissue sections were incubated with a monoclonal antibody to PCNA (DAKO, lot #016, PAI No. A1723) and reagents required for the avidin-biotin peroxidase (ABC Kit, lot #PK-6100, PAI No. K 324) method for the detection of the antigen-antibody complex. PCNA expression in cells was localized by the chromagen 3,3'-diaminobenzidine (DAB; Sigma Chemical Co., lot #18H8201). Tissue sections were counterstained with hematoxylin. 000365 229 Covance 6329-222 3M T-6295.6 Cell Proliferation M easurem ents Final Cell Proliferation Report Covance Study Number 6329-222 Page 3 The percentage of proliferating cells (proliferating index, PI) was determined by scoring at least 3000 hepatocytes in 10 fields of liver, at least 500 Leydig cells of the testes (males only), and at least 2000 acinar cells of the pancreas per animal. A negative control slide was included in the staining run and consisted of study tissue that was not incubated with the primary antibody. For cell proliferation evaluations, slides were first perused at low magnification (100X) to judge quality of staining, processing and sectioning, potential patterns of cellular proliferation, and histomorphologic changes. Cell proliferation was then quantified at higher magnification (200X for liver, and 400X for testes and pancreas) as described above. Histomorphology was further assessed by evaluating the H&E slide prepared from the same tissue block for each animal evaluated for cell proliferation. Statistical Analysis Due to the small sample size, statistical analysis was not performed. RESULTS Cell Proliferation Individual animal cell proliferation data are presented in Section II (Table 1). Cell proliferation in the liver, testes and pancreas of control and test material-treated animals was similar. H istopathology Sections from the same tissue blocks used for preparation of PCNA-stained slides were stained with hematoxylin and eosin (H&E) for histopathologic evaluation to facilitate the interpretation of the immunostained slides. Individual animal findings are presented in Appendix I. Single sections of liver, pancreas, and testes from 6 young adult male monkeys and single sections of liver and pancreas from 6 young adult female monkeys representing control, low-dose (0.02 mg/kg/day), and high-dose (2.0 mg/kg/day) treatment groups were evaluated histologically. Each tissue section was stained with hematoxylin and eosin. The purpose of this evaluation was to determine whether or not morphologic changes were occurring in these tissues which may alter or confound the specificity of the PCNA staining observed in these animals. The results showed that no significant changes were observed in either the liver, pancreas, or testicular tissues from male monkeys or the liver and pancreas tissues from female monkeys, which would alter the interpretation of the PCNA staining in this study. 000366 230 Covance 6329-222 3M T-6295.6 DISCUSSION Final Cell Proliferation Report Covance Study Number 6329-222 Page 4 In the present study, cell proliferation was measured within the liver, testes and pancreas of male monkeys, and the liver and pancreas of female monkeys from control (0 mg/kg/day), low dose (0.02 mg/kg/day), and high dose (2.0 mg/kg/day) groups after four weeks on study to determine an estimated maximun-tolerated dose and lower dose levels to be used in a chronic toxicity study and to assess the effect of the test material on critical enzyme levels, hormones, and other selected biological parameters, to include cell proliferation. There did not appear to be a cell proliferative response to the test material in the liver, testes or pancreas, as determined by proliferating indices which were similar in all animals. SUMMARY In the present study, cell proliferation, as determined by measuring the proliferating index, was not increased in the liver, testes or pancreas of monkeys. 000367 231 Covance 6329-222 3M T-6295.6 Fual Cell Proliferation Report Covance Study Number 6329-222 II. TABLE 000368 COVANCE STUDY NO. 6329-222 TABLE 1. CELL PROUFERATION IN MONKEYS Covance 6329-222 3M T-6295.6 Group 1- 0 mg/kg/day (Control) 1- 0 mg/kg/day (Control) 2 - 0.02 mg/kg/day (Low dose) 2 - 0.02 mg/kg/day (Low dose) 2 - 0.02 mg/kg/day (Low dose) 3 - 2.0 mg/kg/dav (High dose) 1- 0 mg/kg/day (Control) 1- 0 mg/kg/day (Control) 2 - 0.02 mg/kg/day (Low dose) 2 - 0.02 mg/kg/day (Low dose) 2 - 0.02 mg/kg/day (Low dose) 3 - 2.0 mg/kg/day (High dose) Proliferating Animal Index Sex Number Uver M 105349 0.025% M 105350 0.087% M 105344 0.056% M 105347 0.298% M 105346 0.106% M 105345 0.026% F 105366 0.000% F 105367 F 105364 0.086% 0.000% F 105365 F 105370 F 105369 0.083% 0.143% 0.000% Proliferating Index Tectes 20.3% 22.9% 18.5% 33.6% 16.5% 17.6% NA NA NA NA NA NA proliferating Index Pancreas 62.30% 50.30% 66.30% 55.50% 63.90% 44.90% 50.20% 65.60% 39.60% 43.60% 60.20% 40.90% 233 ooo Covance 6329-222 3M T-6295.6 Final Cell Proliferation Report Covance Study Number 6329-222 APPENDIX I 0003/0 234 Covance 6329-222 3M T-6295.6 Covance Study Number 6329-222 Individual A nim al H istopathology Findings M ale M onkeys 4-w eek C apsule Toxicity Study Animal Number 105349 105350 105344 105347 105348 105345 Liver Testes Pancreas No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings Individual A nim al H istopathology Findings - Fem ale M onkeys 4-w eek C apsule Toxicity Study 105366 105367 105364 105365 105370 105369 Animal Number Liver No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings Pancreas No significant findings No significant findings No significant findings No significant findings No significant findings No significant findings 000371 Covance 6329-222 ____ 3M T-6295.6 ID. SIGNATURE PAGE 4-Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys (Covance Study Number 6329-222) Submitted by: PAI Project Manager: Sandra R. Eldridge, Ph.D. PAI Project Pathologist: Date 000372 236 Covance 6329-222 3M T-6295.6 Final Cell Proliferation Report Covance Study Number 6329-222 IV. QUALITY ASSURANCE STATEMENT 003V Pathology Associates International A Company ofSoane Applications krtemationtl Corporation Covance 6329-222 3M T-6295.6 Cell Proliferation Report 4-W eek Capsule Toxicity Study W ith Perfluorooctane Sulfonic Acid Potassium Salt (PFOS; T-6295) in Cynomolgus Monkeys Covance Study Number: 6329-222 QUALITY ASSURANCE STATEMENT This cell proliferation project has been inspected and audited by the PAI Quality Assurance Unit (QAU) as required by the Good Laboratory Practice (GLP) regulations promulgated by the U.S. Environmental Protection Agency (TSCA). The cell proliferation report is an accurate reflection of the recorded data. The following table is a record o f the inspections/audits performed and reported by the QAU. Date of Inspection Phase Inspected Date Findings Reported to PAI Management/Studv Pathologist 06/24/98 08/27;09/08,10/98 08/27;09/08,10/98 11/11/99 Tissue Labeling Study Data and Supporting Documentation Draft Cell Proliferation Report Final Cell Proliferation Report 06/26/98 09/10/98 09/10/98 11/11/99 ~3r. t Karen E. Butler Quality Assurance Specialist 15 Worman's Mill Court, Suite I Frederick, Maryland 21701 (301) 663-1644 (301)663-8994 FAX U00374 238