Document nmgjr9740RzvQpEmDDYB7qEr8

FINAL REPORT PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 FINAL REPORT DATE: 31 JULY 2003 PROTOCOL 418-028 - ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 TABLE OF CONTENTS SUBJECT 1. SUMMARY AND CONCLUSION 1.1. Methods 1.2. Results 1.3. Conclusion 2. DESCRIPTION OF TEST PROCEDURES 2.1. Conduct of Study 2.2. Test Substance Information 2.3. Vehicle Information 2.4. Test Substance Preparation and Storage Conditions 2.5. Test System 2.6. Husbandry 2.7. Methods 3. RESULTS - Male Rats 3.1. Mortality, Clinical and Necropsy Observations 3.2. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight and Brain Weight PAGE 1-1 1-1 1-3 1-6 2-1 2-1 2-4 2-4 2-5 2-6 2-7 2-10 3-1 3-1 3-2 ii SUBJECT PAGE 3.3. Hematology and Clinical Chemistry 3-2 3.4. Body Weights and Body Weight Changes 3-3 3.5. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values 3-3 3.6. Mating and Fertility 3-3 3.7. Functional Observational Battery 3-3 3.8. Motor Activity 3-4 3.9, Sperm 3-4 4. RESULTS - Female Rats 4-1 4.1. Mortality, Clinical and Necropsy Observations 4-1 4.2. Terminal Body Weights, Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight and Brain Weight and Primordial Follicle Counts 4-2 4.3. Hematology and Clinical Chemistry 4-2 4.4. Body Weights and Body Weight Changes 4-3 4.5 Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values 4-3 4.6. Estrous Cycling, Mating and Fertility 4-4 4.7. Functional Observational Battery 4-4 4.8. Motor Activity 4-4 4.9. Natural Delivery and Litter Observations 4-5 4.10. Pup Clinical and Necropsy Observations 4-5 4.11. Pup Liver Weight and Ratio of Liver Weight to Terminal Body Weight 4-5 REFERENCES 4-6 APPENDIX A - REPORT FIGURES Figure 1. Fo Generation Male Rats A-1 . 111 SUBJECT PAGE Figure 2. Figure 3. Fo Generation Female Rats A-2 Motor Activity - Number of Movements - Fo Generation Male Rats A-3 Figure 4. Motor Activity - Time Spent in Movement - Fo Generation Male Rats A-4 Figure 5. Motor Activity - Number of Movements - Fo Generation Female Rats A-5 Figure 6. Motor Activity - Time Spent in Movement - Fo Generation Female Rats A-6 APPENDIX B - REPORT TABLES - Fo GENERATION MALE RATS Table B1. Clinical Observations - Summary - Fo Generation Male Rats B-1 Table B2. Necropsy Observations - Summary - Fo Generation Male Rats B-2 Table B3. Terminal Body Weights and Organ Weights - Summary - Fo Generation Male Rats B-3 Table B4. Ratios (%) of Organ Weight to Terminal Body Weight - Summary - Fo Generation Male Rats B-5 Table B5. Ratios (%) of Organ Weight to Brain Weight - Summary - Fo Generation Male Rats B-7 Table B6. Table B7. Table B8. Table B9. Table B10. Hematology - Summary - Fo Generation Male Rats Clinical Chemistry - Summary - Fo Generation Male Rats Body Weights - Summary - Fo Generation Male Rats Body Weight Changes - Summary - Fo Generation Male Rats Absolute Feed Consumption Values (g/day) - Summary. Fo Generation Male Rats B-9 B-12 B-15 B-16 B-17 Table B 11. Relative Feed Consumption Values (g/kg/day) - Summary - Fo Generation Male Rats B-18 Table B12. Mating and Fertility - Summary - Fo Generation Male Rats B-19 Table B 13. Functional Observational Battery Observations - Summary - Fo Generation Male Rats B-20 iv SUBJECT PAGE Table B14. Motor Activity - Summary - Fo Generation Male Rats B-26 Table B 15. Sperm Motility, Count and Density - Summary - Fo Generation Male Rats B-28 Table B 16. Sperm Morphology - Summary - Fo Generation Male Rats B-29 Table B17. Clinical Observations - Individual Data - Fo Generation Male Rats B-30 Table B 18. Necropsy Observations - Individual Data - Fo Generation Male Rats B-33 Table B 19. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight - Individual Data - Fo Generation Male Rats B-38 Table B20. Organ Weights and Ratios (%) Of Organ Weight to Brain Weight - Individual Data - Fo Generation Male Rats B-48 Table B21. Table B22. Body Weights - Individual Data - Fo Generation Male Rats Feed Consumption Values - Individual Data - Fo Generation Male Rats B-58 B-68 Table B23. Mating and Fertility - Individual Data - Fo Generation Male Rats B-71 Table B24. Functional Observational Battery Observations - Individual Data - Fo Generation Male Rats B-74 Table B25. Motor Activity - Individual Data - Fo Generation Male Rats B-79 Table B26. Sperm Motility, Count and Density - Individual Data - Fo Generation Male Rats B-89 Table B27. Sperm Morphology - Individual Data - Fo Generation Male Rats B-92 APPENDIX C - REPORT TABLES - Fo GENERATION FEMALE RATS Table C 1. Table C2. Table C3. Clinical Observations - Summary - Fo Generation Female Rats C-1 Necropsy Observations - Summary - Fo Generation Female Rats C-4 Terminal Body Weights and Organ Weights and Primordial Follicle Count - Summary - Fo Generation Female Rats C-5 SUBJECT PAGE Table C4. Ratios (%) of Organ Weight to Terminal Body Weight - Summary. - Fo Generation Female Rats C-7 Table C5. Ratios (%) of Organ Weight to Brain Weight - Summary - Fo Generation Female Rats C-8 Table C6. Table C7. Table C8. Hematology - Summary - Fo Generation Female Rats Clinical Chemistry - Summary - Fo Generation Female Rats Body Weights - Precohabitation - Summary - Fo Generation Female Rats C-9 C-12 C- 15 Table C9. Body Weight Changes - Precohabitation - Summary - Fo Generation Female Rats C- 16 Table C10. Maternal Body Weights - Gestation - Summary - Fo Generation Female Rats C- 17 Table C11. Maternal Body Weight Changes - Gestation - Summary - Fo Generation Female Rats C-19 Table C12. Maternal Body Weights - Lactation - Summary - Fo Generation Female Rats C-20 Table C13. Maternal Body Weight Changes - Lactation - Summary - Fo Generation Female Rats C-22 Table C14. Table C15. Table C16. Table C17. Table C18. Absolute Feed Consumption Values (g/day) - Precohabitation - Summary - Fo Generation Female Rats C-23 Relative Feed Consumption Values (g/kg/day) - Precohabitation - Summary - Fo Generation Female Rats C-24 Maternal Absolute Feed Consumption Values (g/day) - Gestation - Summary - Fo Generation Female Rats C-25 Maternal Relative Feed Consumption Values (g/kg/day) - Gestation - Summary - Fo Generation Female Rats C-26 Maternal Absolute Feed Consumption Values (g/day) - Lactation - Summary - Fo Generation Female Rats C-27 vi SUBJECT PAGE Table C19. Table C20. Table C21. Maternal Relative Feed Consumption Values (g/kg/day) - Lactation - Summary - Fo Generation Female Rats C-28 Mating and Fertility, Estrous Cycling and Days in Cohabitation - Summary - Fo Generation Female Rats C-29 Functional Observational Battery - Summary - Fo Generation Female Rats C-31 Table C22. Motor Activity - Summary - Fo Generation Female Rats C-37 Table C23. Natural Delivery Observations - Summary - Fo Generation Female Rats C-39 Table C24. Litter Observations (Naturally Delivered Pups) - Summary - F1 Generation Litters C-40 Table C25. Table C26. Table C27. Table C28. Clinical Observations from Birth to Day 22 Postpartum Summary - F1 Generation Pups Necropsy Observations - Summary - F1 Generation Pups Pup Liver Weights - Summary - F1 Generation Pups Clinical Observations - Individual Data Fo Generation Female Rats C-43 C-44 C-45 C-46 Table C29. Necropsy Observations - Individual Data - Fo Generation Female Rats C-51 Table C30. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight - Individual Data - Fo Generation Female Rats C-55 Table C31. Table C32. Organ Weights and Ratios (%) Of Organ Weight to Brain Weight - Individual Data - Fo Generation Female Rats C-60 Primordial Follicle Count - Individual Data - Fo Generation Female Rats C-65 Table C33. Body Weights - Precohabitation - Individual Data - Fo Generation Female Rats C-67 vii SUBJECT PAGE Table C34. Maternal Body Weights - Presumed Gestation - Individual Data - Fo Generation Female Rats C-70 Table C35. Maternal Body Weights - Lactation - Individual Data - Fo Generation Female Rats C-75 Table C36. Feed Consumption Values - Precohabitation - Individual Data - Fo Generation Female Rats C-80 Table C37. Maternal Feed Consumption Values - Presumed Gestation - Individual Data - Fo Generation Female Rats C-83 Table C38. Maternal Feed Consumption Values - Lactation - Individual Data - Fo Generation Female Rats C-86 Table C39. Mating and Fertility, Estrous Cycling and Days in Cohabitation - Individual Data - Fo Generations Female Rats C-89 Table C40. Functional Observational Battery - Individual Data - Fo Generation Female Rats C-92 Table C41. Table C42. Motor Activity - Individual Data - Fo Generation Female Rats C-97 Natural Delivery, Implantation Sites, and Pup Viability and Sex Individual Data - Fo Generation Female Rats/F1 Generation Litters C-107 Table C43. Pup Body Weight Litter Averages from Birth to Day 22 Postpartum - Individual Data - F1 Generation Litters C-110 Table C44. Pup Body Weights from Birth to Day 22 Postpartum Individual Data - F1 Generation Pups C-113 Table C45. Pup Vital Status and Sex from Birth to Day 22 Postpartum Individual Data - F1 Generation Pups C-125 Table C46. Clinical Observations from Birth to Day 22 Postpartum Individual Data - F1 Generation Pups C-128 Table C47. Necropsy Observations - Individual Data - F1 Generation Pups C-130 Table C48. PUp Liver Weights - Individual Date - F1 Generation Pups APPENDIX D - PROTOCOL AND AMENDMENTS C-136 D-1 to D-52 Vlll It SUBJECT PAGE APPENDIX E - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY E-1 to E-3 APPENDIX F - CERTIFICATE OF ANALYSIS F-1 to F-3 APPENDIX G - ANALYTICAL AND BIOANALYTICAL REPORT G-1 to G-153 APPENDIX H - TEMPERATURE AND RELATIVE HUMIDITY REPORT H- 1 APPENDIX I - POSITIVE CONTROL DATA I-1 to I-4 APPENDIX J - HISTOPATHOLOGY REPORT J-1 to J-105 APPENIDX K - HEMATOLOGY AND CLINICAL CHEMISTRY REPORTS K-1 to K-150 APPENDIX L - STATEMENT OF THE STUDY DIRECTOR L-1 APPENIDIX M - QUALITY ASSURANCE STATEMENT M-1 to M-2 ix 418-028:PAGE 1-1 TITLE: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST ARGUS RESEARCH PROTOCOL NUMBER: 418-028 SPONSOR'S STUDY NUMBER: T-7706.1 1. SUMMARY AND CONCLUSION 1.1. Methods a Seventy-five Crl:CD(SD)IGS VAF/Plus rats per sex were assigned to five dosage groups (Groups I through V), 15 rats per sex per group. An additional three rats per sex per group were assigned to Groups I through V for toxicokinetic sample collection. The test substance, T-7706 [Perfluorohexane Sulfonate Potassium Salt (PFHS)], or vehicle, aqueous 0.5% carboxymethylcellulose (CMC), was administered via gavage to male rats once daily beginning 14 days before cohabitation and continuing through the day before sacrifice, after completion of the cohabitation period, after a minimum of 42 days of administration, and to female rats once daily beginning 14 days before cohabitation and continuing through the day before sacrifice, day 21 of lactation (DL 21) or day 25 of presumed gestation (DG 25, rats that did not deliver a litter). Dosages were 0, 0.3, 1, 3 and 10 mg/kg/day. The dosage volume, 10 mL/kg, was adjusted daily on the basis of the individual body weights recorded before intubation. F1 generation pups were not directly administered the test substance or vehicle. Within each dosage group, rats were assigned to cohabitation, one male rat per female rat. Rats were observed for viability at least twice each day of the study. Observations for clinical signs of effects of the test substance and deaths were made on the first day of dosage at approximately hourly intervals for the first four hours and at the end of the normal working day. Observations for clinical signs of effects of the test substance and deaths were made on subsequent days daily before dosage and approximately 60 + 10 minutes after dosage administration and on the day of sacrifice. Once before the first dosage and at least once weekly thereafter, detailed clinical observations were conducted for all male and female rats assigned to the main study. a. Detailed descriptions of all procedures used in the conduct of this study are provided in the appropriate sections of this report and in APPENDIX D (PROTOCOL AND AMENDMENTS). 418-028:PAGE 1-2 I Body weights were recorded daily during the dosage period and at sacrifice. Feed consumption values for male rats assigned to the main study were recorded weekly during the dosage period. Feed consumption values for female rats assigned to the main study were recorded weekly to cohabitation, on DGs 0, 7, 10, 12, 15, 18, 20 and 25 (if necessary) and on DLs 1, 5, 8 and 15. During cohabitation, individual values were not recorded or tabulated. Estrous cycling was evaluated in rats assigned to the main study by examination of vaginal cytology beginning with the day after the first administration and then until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ during the cohabitation period. Estrous cycling was evaluated in rats assigned to the toxicokinetic study during the cohabitation period until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ. Female rats were evaluated for adverse clinical signs observed during parturition, duration of gestation, litter sizes and pup viability at birth. Maternal behavior was evaluated on DLs 1, 5, 8, 15 and 22. Shortly before scheduled sacrifice, a functional observational battery (FOB) was conducted and motor activity was evaluated on 10 male and 10 female rats per group. On days 14 and 42 of study, blood samples were collected from each male rat assigned to the toxicokinefic sample collection portion of the study and on day 14 of study and DG 21, blood samples were collected from each female rat assigned to the toxicokinefic sample collection portion of the study. Each litter was evaluated for viability at least twice daily. The pups in each litter were counted once daily. Clinical observations were recorded once daily. Pup body weights were recorded on DLs 1, 8, 15 and 22. Male and female rats assigned to the toxicokinefic study were sacrificed on day 42 of study and DG 21, respectively. Liver weights were recorded. The median liver lobe was shipped for analysis. Blood samples were collected from each fetus and pooled by litter and serum was shipped for analysis. The liver from each fetus was collected, pooled per litter and shipped for analysis. The number of implantation sites was recorded. Male and female rats assigned to the main study were sacrificed after a minimum of 42 days of dosage and on DL 22, respectively. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number of implantation sites were recorded. Gross lesions were examined histologically. Ten rats per sex per group assigned to functional observational battery and motor activity tests were assigned to histological evaluations. The following organs were individually weighed: liver, kidneys, adrenals, thymus, testes, right epididymis, left epididymis (corpus and caput), seminal vesicles (with and without fluid), prostate, spleen, brain, heart, ovaries and uterus (with cervix). The following tissues or representative samples 418-028:PAGE 1-3 were retained: brain, small and large intestines, lungs, lymph nodes, peripheral nerve. stomach, kidneys, spleen, thymus, trachea, urinary bladder, spinal cord, liver, adrenals, heart, thyroid/parathyroid, bone marrow, testes, prostate, seminal vesicles, ovaries, uterus, vagina, mammary gland (female rats only) and gross lesions. Histological examination of retained tissues, including reproductive organs, was conducted for the assigned ten rats per sex from the control and high dosage groups. A quantitative evaluation of primordial follicles was conducted for Fo generation female rats. Sperm evaluations (concentration, motility and morphology) were performed for 10 male rats in each dosage group. At scheduled sacrifice, blood samples were collected from the 10 male and 10 female rats per group assigned to hematology and clinical chemistry sample collection. The following hematologic parameters were evaluated: erythrocyte count, hematocrit, hemoglobin, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, mean corpuscular volume, total leukocyte count, differential leukocyte count, platelet count, mean platelet volume and cell morphology. Two blood smear slides were prepared for measurements of differentia/leukocyte count. Plasma samples evaluated for prothrombin time and activated partial thromboplastin time. Sera samples were evaluated for total protein, triglycerides, albumin, globulin, albumin/globulin ratio, glucose, cholesterol, total bilirubin, urea nitrogen, creatinine, creatinine kinase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, calcium, phosphorus, sodium, potassium and chloride. On DL 22, pups were sacrificed and examined for gross lesions. Necropsy included a single cross-section of the head at the level of the frontal-parietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Blood samples were collected from five pups per sex per litter from the 10 female rats per group selected for FOB and motor activity assessment, blood sample collection for hematology and clinical chemistry and histological evaluations. Sera was shipped for analysis. The liver from each selected pup was weighed. 1.2. Results 1.2.1. Male Rats All male rats survived to scheduled sacrifice and all clinical and necropsy observations were considered unrelated to the test substance. Body weight gains were significantly reduced in the 0.3, 3 and 10 mg/kg/day dosage groups on study days (DSs) 29 to 36. Significantly reduced body weight gain occurred in the 0.3, 1, 3 and 10 mg/kg/day dosage groups on study days DS 29 to termination. Body weight gain was also significantly reduced for the 10 mg/kg/day dosage group for the entire dosage period. Absolute and relative feed consumption values for the male rats were unaffected by dosages of the test substance as high as 10 mg/kg/day. 418-028:PAGE 1-4 Terminal body weights of the male rats were slightly reduced in the 10 mg/k_day dosage group. The absolute weights of the liver, the ratios of the liver weights to tern_nal body weights and ratios of the liver weights to brain weight were significantly increased in the 3 and 10 mg/kg/day dosage groups. The ratio of the heart weights to brain weight was significantly decreased in the 10 mg/kg/day dosage group. Hemoglobin concentrations were significantly decreased in the 1, 3 and 10 mg/kg/day dosage groups and average values for red blood cells and hematocrit were significantly decreased in the 3 and 10 mg/kg/day dosage groups. Prothrombin time was sigmficantly increased in the 0.3, 3 and 10 mg/kg/day dosage groups. Average values for cholesterol were significantly decreased in the 0.3, 1, 3 and 10 mg/kg/day dosage groups and the average values for triglycerides was significantly decreased in the 10 mg/kg/day dosage group. Albumin, blood urea nitrogen, alkaline phosphatase, calcium and albumin/globulin ratio levels were significantly increased in the 10 mg]kg/day dosage group. Treatment-related microscopic changes were observed in the liver and thyroid gland of male rats in the 3 and 10 mg/kg/day dosage groups. The treatment-related microscopic changes in the liver consisted of minimal to moderate enlargement of centrilobular hepatocytes and in the thyroid gland as an increased incidence of hypertrophy of follicular cells and hyperplasia. These microscopic changes are consistent with the known effects of compounds that cause microsomal enzyme induction where the hepatocellular hypertrophy results in a compensatory hypertrophy and hyperplasia of the thyroid due to increased plasma turnover of thyroxine and associated stimulation of thyroid-stimulating hormone in rats (1). There were no treatment-related microscopic changes observed in the liver of the F1 generation pups from the dams given up to 10 mg/kg/day of the test substance. All mating and fertility parameters were unaffected by dosages of the test substance as high as 10 mg/kg/day. There were no other statistically significant or biologically important differences among the four dosage groups in the measures of the functional observational battery (FOB) or motor activity on DSs 36 through 39. Sperm motility was unaffected by dosages of the test substance as high as 10 mg/kg/day. The sperm count and sperm density were comparable among the five dosage groups. 1.2.2. Female Rats All female rats survived to scheduled sacrifice and all clinical and necropsy observations were considered unrelated to the test substance. Body weights, body weight gains, and absolute and relative feed consumption values were comparable and did not differ significantly during the precohabitation, gestation or lactation periods at dosages of the test substance up to 10 mg/kg/day. Terminal body weights, absolute and relative weights of the reproductive organs, brain, liver, left and right kidneys and adrenals, spleen, thymus and heart of the female rats were 418-028:PAGE 1-5 comparable among dosage groups. Average primordial follicle counts for the 10 mg/kg/day dosage group were comparable to the control group. No treatment-related microscopic changes were observed in any of the female rats administered up to 10 mg/kg/day of the test substance. There were no treatment-related microscopic changes observed in the liver of the F1 generation pups from the dams given up to 10 mg/kg/day of the test substance. Dosages as high as 10 mg/kg/day did not affect any hematology or clinical chemistry values evaluated. The average numbers of estrous stages per 13 days were comparable among the five dosage groups. All mating and fertility parameters, including the gestation index, viability index and lactation indices were unaffected by dosages of the test substance as high as 10 mg/kg/day. There were no statistically significant or biologically important differences among the five dosage groups in the measures of the functional observational battery (FOB) or motor activity on DL 17. All pregnant dams delivered a litter of one or more live pups. Values for the numbers of dams delivering litters, the duration of gestation, averages for implantation sites per delivered litter, the numbers of dams with stillborn pups, the numbers of dams with no liveborn pups, dams with all pups dying, were comparable among the five dosage groups. The number of pups surviving per litter, pup sex ratios, litter size and pup body weights per litter were comparable among the five dosage groups. No clinical or necropsy observations in the F1 generation pups were attributable to dosages of the test substance as high as 10 mg/kg/day. F1 generation male and female pup terminal body weights, absolute liver weight and ratio of liver weight to terminal body weight were comparable across all five dosage groups. 418-028:PAGE 1-6 1.3. Conclusion On the basis of these data, the maternal no-observable-adverse-effect-level (NOAEL) for T-7706 [Perfluorohexane Sulfonate Potassium Salt] is greater than 10 m Wkg/day (the 10 mg/kg/day dosage caused no deaths, adverse clinical or necropsy/pathology observations, changes in body weight, feed consumption, or hematology or clinical chemistry values throughout precohabitation, gestation or lactation. The paternal NOAEL is less than 0.3 mg/kg/day (the 0.3 and 1 mg/kJday dosages caused significant differences in body weight gain, hematology and clinical chemistry values and the 3 and 10 mg/kg/day dosages caused significant changes in absolute and relative organ weights and microscopic changes in the liver and thyroid gland). The reproductive NOAEL is greater than 10 mg/kg/day (the 10 mg/kg/day dosage had no effect on the durations of gestation and parturition or any mating and fertility parameters. There was no effect on the sperm parameters in the male rats). The NOAEL for viability and growth in the offspring is also greater than 10 mg/kg/day (dosages of 10 mg/kg/day had no effect on perinatal mortality, clinical or necropsy observations or body or liver weights in the F1 generation offspring). ................ Alan M. Hoberman, Ph.D., DABT Date Director of Research Study Director R_y_nc_ndG. Yor_ Ph.D l, DABT Date Assdei_ateDirectob, oLR_earch Study Director 418-028:PAGE 2-1 I 2. DESCRIPTION OF TEST PROCEDURES 2.1. Conduct of Study 2.1.1. Sponsor 3M Corporate Toxicology, 3M Center, Building 220-2E-02, St. Paul, Minnesota 55144-1000 2.1.2. Testing Facility Argus Research, 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 2.1.3. 418-028 Study Number 2.1.4. Sponsor's Study Number T-7706.1 2.1.5. Purpose of the Study The purpose of this study was to provide information on the possible health hazards that may result from repeated exposure of Crl:CD(SD)IGS BR VAF/Plus male and female rats to a test substance beginning before cohabitation, through mating and continuing for at least 42 days (male rats) or through parturition until day 21 of lactation (female rats). This repeated dose study incorporated a reproduction/developmental toxicity screening test that can be used to provide initial information on possible effects on male and female reproductive performance (e.g., gonadal function, mating behavior, conception, development of the conceptus and parturition). The study also placed emphasis on neurological effects as a specific endpoint and should identify the neurotoxic potential of a test substance, which may warrant further in-depth investigation. Because of the selectivity of the endpoints and the short duration of the study, the screening test did not provide evidence for definitive claims of no reproductive/ developmental effects. In particular, it offered only limited means of detecting postnatal manifestations of prenatal exposure or effects that may be induced during postnatal exposure. 2.1.6. Study Design The requirements of the Organisation for Economic Co-operation and Development (OECD) (2)were used as the basis for study design. 2.1.7. Regulatory Compliance This study was conducted in compliance with Good Laboratory Practice (GLP) regulations of the OECD(3),U.S. Food and Drug Administration (FDA) (4_,the Japanese 418-028:PAGE 2-2 - .(51 Ministry of Health and Welfare (MHW) . There were no deviations from the GLP regulations that affected the quality or integrity of the study. Quality Assurance Unit findings derived from the inspections during the conduct of this study are documented and have been provided to the Study Director and the Testing Facility Management. 2.1.8. Ownership of the Study The Sponsor owns the study. All raw data, analyses, reports and preserved tissues are the property of the Sponsor. 2.1.9. Study Monitor John Butenhoff, Ph.D., CIH, DABT 2.1.10. Study Director Raymond G. York, Ph.D., DABT (Associate Director of Research) Address as cited previously for Testing Facility. 2.1.11. Technical Performance John F. Barnett, B.S. (Director of Laboratory Operations) Joseph W. Lech, B.S. (Senior Research Associate) Mary P. Howard, B.S. (Team Leader - General Laboratory) Jaclyn S. Fox, B.S. (Laboratory Technician) Josette M. Provost, B.S. (Necropsy Laboratory Technician/Fixed Tissue Christopher K. Ruppert, B.S. (Formulation Laboratory Technician) Coordinator) 2.1.12. Report Preparation Raymond G. York, Ph.D., DABT Jo Ann Frazee, M.S. (Study Coordinator) JoAnne M. Conldin, B.S. (Data Management Cristina Petrescu (Report Administrator) Specialist) 2.1.13. Report Review Valerie A. Sharper, M.S. (Director of Study Management) 2.1.14. Date Protocol Signed 26 March 2002 2.1.15. Dates of Technical Performance 2.1.15.1. Male Rats Rat Arrival Dosage Period (14 days before cohabitation and through a 14-day cohabitation period until sacrifice after at least 42 days of dosage) FOB and Motor Activity Evaluation Toxicokinetic Sample Collections DSa 14 DS 42 Scheduled Sacrifice - Toxicokinetic Study Scheduled Sacrifice - Main Study 2.1.15.2. Female Rats Rat Arrival Dosage Period (14 days before cohabitation through DL b 21) Cohabitation Period Male 1 Male 2 Toxicokinetic Sample Collections DS 14 DG c 21 DG 0 DG 25 Sacrifice (Rats that did not deliver a litter) Delivery Period a (DL 1) FOB and Motor Activity Evaluation DL 22 Sacrifice Female Rats and Pups 418-028:PAGE 2-3 26 MAR 02 01 APR 02 - 16 MAY 02 06 MAY 02 - 09 MAY 02 14 APR 02 12 MAY 02 12 MAY 02 13 MAY 02 - 17 MAY 02 26 MAR 02 01 APR 02 - 09 JUN 02 14 APR 02 PM - 21 APR 02 AM 21 APR 02 PM - 28 APR 02 AM 14 APR 02 06 MAY 02 - 19 MAY 02 15 APR 02 - 28 APR 02 10 MAY 02 - 23 MAY 02 07 MAY 02 - 20 MAY 02 23 MAY 02 - 26 MAY 02 28 MAY 02 - 10 JUN 02 a. DS is an abbreviation used for day of study. b. DL is an abbreviation used for day of lactation/postpartum. c. DG is an abbreviation used for day of (presumed) gestation. d. The day of birth is designated lactation day 0 (postpartum day 0) in the Health Effects Test Guidelines - Reproduction and Fertility Effects (Office of Prevention, Pesticides and Toxic Substances 870.3800, August, 1998) and in the OECD Guideline for the Testing of Chemicals - Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test (Section 4, No. 422, 22 March 1966). This same day is designated day 1 postpartum (day 1 of lactation) in the Standard Operating Procedures of the Testing Facility. Throughout this study, the day of birth was designated as day 1 postpartum (day 1 of lactation) and all subsequent ages of the F1 generation rats and days of the lactation period were determined and cited accordingly. 418-028:PAGE 2-4 2.1.16. Records Maintained The original report, raw data and reserve samples of each lot of bulk test substance and bulk vehicle components are retained in the archives of Argus Research. Any preserved tissues are retained in the archives of the Testing Facility for one year after the mailing of the draft final report, after which time the Sponsor will decide their final disposition. All unused prepared formulations were discarded at the Testing Facility. All remaining bulk test substance was returned to the Sponsor. 2.2. Test Substance Information 2.2.1. Description T-7706 [Perfluorohexane Sulfonate Potassium Salt (PFHS)] - a white powder 2.2.2. Date Received and Storage Conditions The test substance was received on 11 March 2002 and stored at room temperature. 2.2.3. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-filtered mask, safety goggles or safety glasses and a face-shield) were taken during formulation preparation and dosage. A half-face respirator was worn during formulation preparation. 2.2.4. Analysis of Purity Information to document or certify the identity, composition, method of synthesis, strength and purity of the test substance was provided by the Sponsor to the Testing Facility. A Certificate of Analysis is available in APPENDIX F. 2.3. Vehicle Information 2.3.1. Description Aqueous 0.5% carboxymethylcellulose (CMC) (sodium salt; medium viscosity) prepared using carboxymethylcellulose (sodium salt; medium viscosity), an off-white powder, and reverse osmosis membrane processed deionized water (R.O. deionizcd water). 2.3.2. Lot Number 120K0252 2.3.3. Date Received and Storage Conditions The carboxymethylcellulose was received from Sigma Chemical Co., St. Louis, Missouri, on 11 September 2001 and stored at room temperature. R.O. deionized water is available from a continuous source at the Testing Facility and is maintained at room temperature. 418-028:PAGE 2-5 2.3.4. Special Handling Instructions Standard safety precautions (use of protective clothing, gloves, dust-mist/HEPA-fi/,tered mask, safety goggles or safety glasses and a face-shield) were taken when handling the vehicle components and prepared vehicle. 2.3.5. Analysis of Purity Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the vehicle that would have interfered with the results of this study. The expiration date for the carboxymethylcellulose is September 2005. 2.4. Test Substance Preparation and Storage Conditions Formulations were prepared weekly at the Testing Facility. Prepared test substance and vehicle formulations were stored refrigerated (2C to 8C). 2.4.1. Sample Information Shipped To/ Date Storage Shipping Date Sample T_,pe Bulk Test Substancea Size Retained 1g 09 JUN 02 Conditions Room temperature Conditions Exygenb/ Ambient conditions Shipped 10JUN 02 Homogeneityc (all levels) Concentrationd (all levels) Stabilityg Bulk Test Substance Reserve 2 mL 2 mL 2 mL 1g 29 MAR 02 24 MAY 02e 07 JUN 02f 29 MAR 02 10 JUN 02 Refrigerated Refrigerated Refrigerated Room temperature Sponsor/ Refrigerated Exygenb/ Refrigerated Sponsor/ Refrigerated Testing Facility Archives 01 APR 02 28 MAY 02 10JUN 02 01 APR 02 10 JUN 02 Vehicle Components Reserve Room temperature Testing Facility Archives Carboxymethylcellulose 1g 10 JUN 02 R.O. deionized water 5 mE 10 JUN 02 10 JUN 02 10 JUN 02 a. A sample of the test article was retained on the last day of treatment and shipped for anal_'sis. b. Exygen Research, State College, Pennsylvania. c. Quadruplicate samples were taken from the top, middle and bottom of each concentration on the first day of preparation. Two samples from each quadruplicate set were shipped for analysis. The remaining samples were retained at the Testing Facility as backup samples. d. Quadruplicate samples were taken from each concentration on the last day of preparation. Two samples from each quadruplicate set were shipped for analysis. The remaining samples were retained at the Testing Facility as backup samples. e. Samples for 0.03, 0.1 and 1mg/mL concentrations only. f. Samples for 0 and 0.3 mg/mL concentrations only. g. Two sets of duplicate samples from each concentration were taken on the first day of preparation. One sample of each duplicate set was shipped on the day of preparation. These samples were analyzed as soon as possible after preparation and ten days after the first analysis. The remaining samples were retained at the Testing Facility as backup samples. 418-028:PAGE 2-6 i 2.4.2. Analytical Results Results of the analytical analyses are available in APPENDIX G. 2.5. Test System 2.5.1. Rat Species 2.5.2. Strain Crl:CD(SD)IGS VAF/Plus 2.5.3. Supplier (Source) Charles River Laboratories, Inc. Male Rats - St. Constant, Quebec, CANADA Female Rats - Raleigh, North Carolina 2.5.4. Sex Male and Female 2.5.5. Rationale for Test System The Crl:CD(SD)IGS BR VAF/Plus rat was selected as the Test System because: 1) it is one mammalian species accepted for use in toxicity studies and it has been widely used throughout industry; 2) this strain of rat has been demonstrated to be sensitive to reproductive and developmental toxins; and 3) historical data and experience exist at the Testing Facility (68). 2.5.6. Test System Data 2.5.6.1. Male Rats Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) the Day after Arrival Weight (g) at Study Assignment 2.5.6.2. Female Rats 100 20 JAN 02 66 days 277 - 318 305 - 348 Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) the Day after Arrival Weight (g) at Study Assignment 100 21 JAN 02 65 days 195 - 227 207 - 230 418-028:PAGE 2-7 2.5.7. Method of Randomization Upon arrival, the male and female rats were assigned to individual housing on the basis of computer-generated random units. During an acclimation period of at least five days, male and female rats were selected for study on the basis of physical appearance and body weights recorded during acclimation. The ratswere assigned to five dosage groups (Groups I through V), 15 rats per sex per group, using a computer-generated (weightordered) randomization procedure. An additional three rats per sex per group were assigned to Groups I through V for toxicokinetic sample collection. At study initiation, the weight variation of the rats did not exceed _ 20% of the mean weight of each sex. Litters were not culled during the lactation period because random selection of pups for culling could have resulted in potential biases in pup viabilities and body weight gains over this period. Within each dosage group, consecutive order was used to assign the first 10 male and the first 10 female Fo generation rats to a functional observational battery (FOB) and motor activity assessment, blood sample collection for clinical chemistry and hematology and histological evaluations. On DL 22, a table of random units was used to select five male and five female pups per litter for blood sample and liver collection. These pups were only selected from the 10 dams selected for FOB, motor activity, clinical chemistry and hematology and histological evaluation. 2.5.8. System of Identification Male and female rats were assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study before administration of the first dosage. Rats were permanently identified using a Monel self-piercing ear tag (Gey Band and Tag Co., Inc., No. MSPT 20101). Cage tags were marked with the study number, permanent rat number, sex, test substance identification, generation, dosage group and dosage level. Pups were not individually identified during lactation; all parameters were evaluated in terms of the litter. 2.6. Husbandry 2.6.1. Research Facility Registration USDA Registration No. 14-R-0144 under the Animal Welfare Act, 7 U.S.C. 2131 et seq. 2.6.2. Study Room The study room was maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum of ten changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters. Room temperature and humidity 418-028:PAGE 2-8 were monitored constantly throughout the study. Room temperature was targeted at 64F to 79F (18C to 26C); relative humidity was targeted at 30% to 70% _. 2.6.3. Housing Rats were individually housed in stainless steel, wire-bottomed cages except during cohabitation and postpartum periods. During cohabitation, each pair of male and female rats was housed in the male rat's cage. Beginning no later than DG 20, Fo generation female rats were individually housed in nesting boxes until they either naturally delivered litters or were sacrificed (DG 25). Each dam and delivered litter was housed in a common nesting box during the postpartum period. All cage sizes and housing conditions were in compliance with the Guide for the Care and Use of Laboratory Animals (9). Argus Research is an AAALAC-accredited facility. 2.6.4. Lighting An automatically-controlled fluorescent light cycle was maintained at 12-hours light: 12-hours dark, with each dark period beginning at 1900 hours EST. 2.6.5. Sanitization Cage pan liners were changed at least three times weekly. Cages were changed approximately every other week. Bedding was changed as often as necessary to keep the rats dry and clean. 2.6.6. Feed Rats were given ad libitum access to Certified Rodent Diet#5002 (PMI Nutrition International, Inc., St. Louis, Missouri) in individual feeders. Feed was removed the evening prior to the scheduled sacrifice b. 2.6.7. Feed Analysis Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration for certified feed or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the feed at concentrations that would have interfered with the results of this study. a. See APPENDIX H (TEMPERATURE AND RELATIVE HUMIDITY REPORT). b. See APPENDIX E (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), item 1. 418-028:PAGE 2-9 2.6.8. Water Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad libitum from individual water bottles attached to the cages and/or from an automatic watering access system. Chlorine was added to the processed water as a bacteriostat. 2.6.9. Water Analysis The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, Pennsylvania) and monthly for possible bacterial contamination (Analytical Laboratories, Inc., Chalfont, Pennsylvania). Copies of the results of the water analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the water that would have interfered with the results of this study. 2.6.10. Bedding Material Bed-o'cobs bedding (The Andersons Industrial Products Group, Maumee, Ohio) was used as the nesting material. 2.6.11. Bedding Analysis Analyses for possible contamination are conducted semi-annually. Copies of the restults of the bedding analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the bedding that would have interfered with the results of this study. 418-028:PAGE 2-10 2.7. 2.7.1. Methods Dosage Administration Dosage Group I Dosage a Concentration _mg/kg/day) (mg/mL) 0 0 Dosage Volume (mlJkg) 10 Number of Rats per Sex 15 + 3 b Assigned Numbers Male Rats Main Study: 19109, 19115, 19116, 19119, 19122, 19125, 19131, 19132, 19134, 19135, 19143, 19151, 19157, 19161, 19167 Toxicokinetic Study: 19176- 19178 Female Rats Main Study: 19010, 19012, 19019. 19021, 19023, 19041, 19042, 19044, 19050, 19053. 19065, 19068, 19072, 19074, 19075 Toxicokinetic Study: 19076- 19078 II 0.3 0.03 10 15 + 3 b Main Study: Main Study: 19102, 19106, 19108, 19110, 19004, 19009, 19016. 19018, 19113, 19120, 19129, 19136, 19026, 19036, 19037, 19043, 19138, 19139, 19147, 19153, 19047, 19048, 19052, 19055, 19164, 19165, 19171 19061, 19067, 19071 Toxicokinetic Study: 19179- 19181 Toxicokinetic Study: 19079 - 19081 II] 1 0.1 10 15 + 3 b Main Study: Main Study: 19101, 19105, 19107, 19112, 19003, 19007, 19008, 19013, 19114, 19121, 19123, 19130, 19014, 19015, 19017, 19024, 19137, 19146, 19155, 19159, 19029, 19034, 19038, 19056, 19172, 19173, 19174 19057, 19060, 19064 Toxicokinetic Study: 19182 - 19184 Toxicokinetic Study: 19082 - 19084 IV 3 0.3 10 15 + 3 b Main Study: Main Study: 19100, 19103, 19104, 19118, 19002, 19005, 19035, 19039, 19133, 19141, 19142, 19144, 19040, 19045, 19046, 19051, 19148, 19150, 19156, 19160, 19054, 19058, 19062, 19063, 19162, 19163, 19166 19066, 19069, 19073 Toxicokinetic Study: 19185 - 19187 Toxicokinetic Study: 19085 - 19087 V 10 1 10 15 + 3 Main Study: Main Study: 19111, 19117, 19124, 19126, 19001, 19006, 19011, 19020, 19127, 19128, 19140, 19145, 19022, 19025, 19027, 19028, 19149, 19152, 19154, 19158, 19030, 19031, 19032, 19033, 19168, 19169, 19170 19049, 19059, 19070 Toxicokinetic Study: 19188 - 19190 Toxicokinetic Study: 19088 - 19090 a. The test substance was considered 100% pure for the purpose of dosage calculations. b. Three additional rats per sex were assigned to toxicokinetic sample collection. 2.7.2. Rationale for Dosage Selection Dosages were selected by the Sponsor based on previous studies conducted with the test substance, taking into account possible differences in sensitivity between pregnant and nonpregnant rats. The highest dosage was expected to cause toxic effects but not mortality or obvious suffering. The descending sequence of the lower dosage levels was selected for the purpose of demonstrating any dosage-related response, with no adverse effects expected at the lowest level. 418-028:PAGE 2-11 m 2.7.3. Route and Rationale for Route of Administration The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered; and 2) it is one of the possible routes for environmental exposure. 2.7.4. 2.7.4.1. Frequency of Administration Fo Generation Rats Male rats were administered the test substance or the vehicle once daily beginning 14 days before cohabitation (maximum 14 days) and continuing through the day before sacrifice, after completion of the cohabitation period, after a minimum of 42 days of administration. Female rats were administered the test substance or the vehicle once daily beginning 14 days before cohabitation (maximum 14 days) and continuing through the day before sacrifice, DL 21 or DG 24 (rats that did not deliver a litter). The dosage volume was adjusted daily on the basis of the individual body weights recorded before intubation. The rats were intubated once daily at approximately the same time each day. The f'u'stday of dosage was designated as DS 1. Dams in the process of delivering pups were not intubated until completion of parturition, in order to preclude possible disruption of maternal behavior and/or cannibalization of pups. Consequently, some dams were not administered one daily dosage during the delivery period. No dam missed more than one daily dosage. 2.7.4.2. F1 Generation Pups F1 generation pups were not directly administered the test substance or vehicle, but may have been possibly exposed to the test substance or vehicle during maternal gestation (in utero exposure) or via maternal milk during the lactation period. 2.7.5. 2.7.5.1. Method of Study Performance Fo Generation Rats Within each dosage group, consecutive order was used to assign rats to cohabitation, one male rat per female rat. The cohabitation period consisted of a maximum of 14 days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug in situ were considered to be DG 0 and assigned to individual housing. Female rats not mated with a male rat within the first seven days of cohabitation were assigned an alternate male rat that had mated (same dosage group) and remained in cohabitation for a maximum of seven additional days. Rats were observed for viability at least twice each day of the study. Rats were examined for clinical observations and general appearance weekly during the acclimation period. Observations for clinical signs of effects of the test substance and deaths were made on the fn'st day of dosage at approximately hourly intervals for the first four hours and at the end of the normal working day. Observations for clinical signs of effects of the test 418-028:PAGE 2-12 substance and deaths were made on subsequent days daily before dosage and approximately 60 _+10 minutes after dosage administration and on the day of sacrificeL Once before the first dosage and at least once weekly thereafter, detailed clinical observations were conducted for all male and female rats assigned to the main study b. These observations were made outside the cage in a standard arena at the same time each day of conduct. Effort was made to ensure that variations in the test conditions were minimal and that observations were conducted by observers unaware of treatment groups. The rats were observed for (but observations were not limited to) the following signs: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g., lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypic behavior (e.g., excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behavior (e.g., self-mutilation, walking backwards) were also recorded. Body weights were recorded weekly during the acclimation period, daily during the dosage period and at sacrifice. Feed consumption values for male rats assigned to the main study were recorded weekly during the dosage period. Feed consumption values for female rats assigned to the main study were recorded weekly to cohabitation, on DGs 0, 7, 10, 12, 15, 18, 20 and 25 (if necessary) and on DLs 1, 5, 8 and 15c. Because pups begin to consume maternal feed on or about DL 15, feed consumption values were not tabulated after DL 15. During cohabitation, when two rats occupied the same cage with one feed jar, replenishment of the feed jars was documented but individual values were not recorded or tabulated. Estrous cycling was evaluated in rats assigned to the main study by examination of vaginal cytology beginning with the day after the first administration and then until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ during the cohabitation period. Estrous cycling was evaluated in rats assigned to the toxicokinetic study during the cohabitation period until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ. Female rats were evaluated for adverse clinical signs observed during parturition, duration of gestation (DG 0 to the day the fu'st pup was observed), litter sizes (all pups delivered) and pup viability at birth. Maternal behavior was evaluated on DLs 1, 5, 8, 15 and 22. Variations from expected maternal behaviorwere recorded, if at!.dwhen present, on all other days of the postpartum period. On one occasion during the course of the study, a functional observational battery (FOB) 13) was conducted on 10 male and 10 female rats per group. For male rats, this assessment was conducted shortly before scheduled sacrifice, but prior to blood sample a. See APPENDIX E, items 2 and 3. b. See APPENDIX E, item 4. c. See APPENDIX E, item 5. 418-028 :PAGE 2- ! 3 collection for hematology and clinical chemistry evaluations. Female rats were tested during the lactation period, shortly before scheduled sacrifice. The FOB evaluation was conducted by an observer unaware of the group assignment of the rat. The following parameters were assessed: 1. Lacrimation, salivation, palpebral closure, prominence of the eye, pupillary reaction to light, piloerection, respiration, and urination and defecation (autonomic functions). 2. Sensorimotor responses to visual, auditory, tactile and painful stimuli (reactivity and sensitivity). 3. Reactions to handling and behavior in the open field (excitability). 4. Gait pattern in the open field, severity of gait abnormalities, air righting reaction and landing foot splay (gait and sensorimotor coordination). 5. Forelimb and hindlimb grip strength. 6. Abnormal clinical signs including but not limited to convulsions, tremors and other unusual behavior, hypotonia or hypertonia, emaciation, dehydration, unkempt appearance and deposits around the eyes, nose or mouth. The ability of this battery to detect the effects of positive control substances has been established (Testing Facility Positive Control Data) and is available in APPENDIX I. Motor activity was evaluated on 10 male and 10 female rats per group once, shortly before scheduled sacrifice, prior to blood sample collection. The movements of each rat were monitored by a passive infrared sensor mounted outside a stainless steel, wirebottomed cage (40.6 x 25.4 x 17.8 cm). Each test session was 1.5 hours in duration with the number of movements and time spent in movement tabulated at each five-minute interval. The apparatus monitored a rack of up to 32 cages and sensors during each session, with each rat tested in the same location on the rack across test sessions. Groups were counterbalanced across testing sessions and cages. Data demonstrating that the test system is capable of detecting increases in activity produced by positive control substances (Testing Facility Positive Control Data) is available in APPENDIX I. On DSs 14 and 42, blood samples (approximately 1 mL each) were collected from each male rat assigned to the toxicokinetie sample coUection portion of the study (three rats per group) and on DS 14 and DG 21, blood samples (approximately 1 mL each) were collected from each female rat assigned to the toxicokinetic sample collection portion of the study (three rats per group). Samples were collected prior to dosage on DS 14. The time of each blood collection was recorded in the raw data. Blood was collected from the orbital sinus on DS 14 of the study and from the vena cava on DS 42 of study (male rats) and DG 21 (female rats). The samples were transferred into EDTA-coated (purple top) tubes and spun in a centrifuge. The resulting serum was transferred into polypropylene 418-028:PAGE 2-14 tubes labeled with the protocol number, Sponsor study number, rat number, sex, group number, dosage level, day of study, collection interval, date of collection, species, generation and storage conditions. All samples were immediately frozen on dry, ice and maintained frozen (<-70C) until shipment to Exygen Research, State College, Pennsylvania, for analysis. Results of these analyses are available in APPENDIX G. 2.7.5.2. F1 Generation Pups Day 1 of lactation (postpartum) was defined as the day of birth and was also the first day on which all pups in a litter were individually weighed (pup body weights were recorded after all pups in a litter were delivered and groomed by the dam). Each litter was evaluated for viability at least twice daily. The pups in each litter were counted once daily. Clinical observations were recorded once daily a. Pups were observed if they were warm and clean, for evidence of a nest and if pups were grouped together and nursing or had milk in stomach. Each pup was examined for general shape of the head, trunk, limbs, tail and presence of anus. Pup body weights were recorded on DLs 1, 8, 15 and 22 (terminal weight). 2.7.6. 2.7.6.1. Gross Necropsy Fo Generation Rats Male and female rats assigned to the toxicokinetic study were sacrificed by carbon dioxide asphyxiation on DS 42 and DG 21, respectively. Blood samples were collected from the rats as previously described. After sacrifice, the liver of each rat was excised and the liver weight was recorded. The median liver lobe was frozen and stored (<-20C) until shipment for analysis b. The fetuses were removed from the uterus and blood samples were collected from each fetus via decapitation. Blood was placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge. The resulting serum was transferred into labeled polypropylene tubes. All samples were immediately frozen on dry ice and maintained frozen (<-70C) until shipment for analysis. The liver from each fetus was collected, pooled per litter and placed into labeled tubes. The samples were frozen and stored (<-20C) until shipment for analysis. The number of implantation sites was recorded. Carcasses were discarded without further evaluation. The livers were shipped to Exygen Research, State College, Pennsylvania, for analysis. Results of the analyses are available in APPENDIX G. Male and female rats assigned to the main study were sacrificed by carbon dioxide asphyxiation following the last dosage administration, after a minimum of 42 days of dosage and on DL 22, respectively. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Gross necropsy included an initial physical examination of external surfaces and all orifices, as well as the cranial, thoracic and abdominal cavities and their contents. Special attention was paid to the organs of the reproductive system. The ovaries and the uterus with cervix of each female rat were weighed, and the a. See APPENDIX E, items 6 and 7. b. See APPENDIX E, item 8. 418-028:PAGE 2-15 ovaries, uterus, vagina and a mammary gland were retained in neutral buffered 10% formalin. The number of implantation sites were recorded. Uteri of apparently nonpregnant rats were examined after being pressed between glass plates to confirm the absence of implantation sites and were retained in neutral buffered 10% formalin. Gross lesions were retained in neutral buffered 10% formalin and examined histologically. Representative photographs of gross lesions are available in the raw data. Tissue trimming and histopathology were performed under the supervision of or by a Board Certified Veterinary Pathologist. Ten rats per sex per group assigned to functional observational battery and motor activity tests were assigned to histological evaluations. The following organs were excised, trimmed and individually weighed as soon as possible after excision to avoid drying: liver, kidneys, adrenals, thymus, testes, right epididymis, left epididymis (corpus and caput), seminal vesicles (with and without fluid), prostate, spleen, brain, heart, ovaries and uterus (with cervix). The following tissues or representative samples were retained in neutral buffered 10% formalin: brain (representative regions including cerebrum, cerebellum, pons), small and large intestines (including Peyer's patches), lungs (perfused with neutral buffered 10% formalin), lymph nodes (submandibular and mediastinal), peripheral nerve (sciatic or tibial), stomach, kidneys, spleen, thymus, trachea, urinary bladder, spinal cord (cervical, thoracic and lumbar), liver, adrenals, heart, thyroid/parathyroid, bone marrow (sternum), testes (fixed in Bouin's solution for 48 to 96 hours before being retained in neutral buffered 10% formalin), prostate, seminal vesicles (with coagulating gland), the remaining portion of the left epididymis (corpus and caput), the right epididymis, ovaries, uterus, vagina, mammary gland (female rats only) and gross lesions. Histological examination of retained tissues, including reproductive organs, was conducted for the assigned ten rats per sex from the control and high dosage groups. A quantitative evaluation of primordial follicles was conducted for Fo generation female rats. Examination included enumeration of number of primordial follicles, which were combined with small growing follicles, for comparison of ovaries of rats assigned to treated and control groups. Tissues that were examined histologically were shipped to Research Pathology Services, Inc., New Britain, Pennsylvania, for evaluation. Results of the histological evaluation are available in APPENDIX J. To assess the potential toxicity of the test substance on the male reproductive system, sperm evaluations were performed for 10 male rats in each dosage group. Sperm concentration and motility were evaluated using computer-assisted sperm analysis (CASA). Motility was evaluated by the Hamilton Thorne IVOS by collection of a sample from the left vas deferens. A homogenate was prepared from the left cauda epididymis for evaluation by the Hamilton Thorne IVOS to determine sperm concentration (sperm per gram of tissue weight). The remaining portion of the left cauda epididymis was used to manually evaluate sperm morphology. Sperm morphology evaluations included the following: 1) determination of the percentage of normal sperm in a sample of at least 200; and 2) qualitative evaluation of abnormal sperm, including such categories as abnormal head, abnormal tail, and abnormal head and tail. 418-028:PAGE 2-16 At scheduled sacrifice, the 10 male and 10 female rats per group assigned to hematology and clinical chemistry sample collection were exsan_inated from the interior vena cava following sacrifice. Rats were fasted overnight before sacrifice a. Approximately 5 mL of blood was collected. The tubes containing the samples were labeled with the protocol number, Sponsor study number, rat number, sex, group number, dosage level, day of study, collection interval, date of collection, species, generation and storage conditions. Approximately 1 mL of blood was collected into EDTA-coated tubes and maintained on wet ice or refrigerated until shipment for analysis of the following hematologic parameters: erythrocyte count (RBC), hematocrit (HCT), hemoglobin (HGB), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), total leukocyte count (WBC), differential leukocyte count, platelet count (PLAT), mean platelet volume (MPV) and cell morphology. Two blood smear slides were prepared at the Testing Facility for each sample for measurements of differential leukocyte count. Approximately 1.8 mL of blood was added to a tube containing 0.2 mL of sodium citrate (0.129 M). The contents were mixed and maintained on wet ice until the tubes were centrifuged (within 30 minutes of the collection time). The resulting plasma was transferred to 2.0 mL polypropylene tubes and immediately frozen. Plasma samples were maintained on dry ice or in a freezer (<-70C) until shipment for measurement of prothrombin time (PT) and activated partial thromboplastin time (APTT). Approximately 2 mL of blood was collected into serum separator tubes and centrifuged. The resulting sera samples were immediately frozen on dry ice and maintained frozen (<-70C) until shipment for analysis of the following parameters: total protein (TP), triglycerides (TRD, albumin (A), globulin (G), albumin/globulin Ratio (A/G), glucose (GLU), cholesterol (CHOL), total bilirubin (TBILI), urea nitrogen (BUN), creatinine (CREAT), creatinine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALK), calcium (CA), phosphorus (PHOS), sodium (NA), potassium (K) and chloride (CL). Samples for hematology and clinical chemistry analyses were shipped to Redfield Laboratories, A Division of CRL-DDS, Redfield, Arkansas. Samples were shipped on dry ice and slides were shipped at ambient conditions. Results of these analyses are available in APPENDIX K. Female rats that did not deliver a litter were sacrificed on DG 25. Gross necropsy, examination and tissue retention were conducted as described above for rats at scheduled sacrifice. Gross lesions were preserved in neutral buffered 10% formalin for possible future evaluation. Representative photographs of gross lesions are available in the raw data. a. See APPENDIX E, item 1. 418-028:PAGE 2-17 2.7.6.2. F1 Generation Pups On DL 22, pups were sacrificed by carbon dioxide asphyxiation and examined for gross lesions. Necropsy included a single cross-section of the head at the level of the frontalparietal suture and examination of the cross-sectioned brain for apparent hydrocephaly. Gross lesions were preserved in neutral buffered 10% formalin for possible future evaluation. Representative photographs of gross lesions are available in the raw data. Blood samples were collected from each selected pup (five per sex per litter from the 10 female rats per group selected for FOB and motor activity assessment, blood sample collection for hematology and clinical chemistry and histological evaluations) from the vena cava. The blood was placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge; the resulting serum was transferred into labeled polypropylene tubes. All samples were immediately frozen on dry ice and maintained frozen (<-70C) until shipment for analysis. The liver from each selected pup was excised and the organ weight recorded a. The median lobe was frozen and stored (<-20C) until shipment for possible analysis. The remaining portion of each liver was retained in neutral buffered 10% formalin for possible histological evaluation. The livers were processed and evaluated histologically as described for the Fo generation rats. Pups that died before initial examination of the litter for pup viability were evaluated for vital status at birth. The lungs were removed and immersed in water. Pups with lungs that sink were identified as stillborn; pups with lungs that float were identified as liveborn, and to have died shortly after birth. Pups found dead were examined for gross lesions and for the cause of death. 2.7.7. Data Collection and Statistical Analyses Data generated during the course of this study were recorded either by hand or using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, the Coulbourn Instruments Passive Infrared Motor Activity System, the Coulbourn Instruments Auditory Startle System, the Coulbourn Instruments Spatial Delayed Alternation System, and/or the passive avoidance software. All data were tabulated, surn_rnarizedand/or statistically analyzed using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microsoft Excel [part of Microsoft Office 97 (version SR-2)] and/or The SAS System (version 6.12). a. See APPENDIX E, item 9. 418-028:PAGE 2-18 Averages and percentages were calculated. Litter values were used where appropriate. The following schematic represents the statistical analyses of the data: I. Parametric TVDe of Test a II. Nonoarametric b A. Bartlett's Testc I II Significant at 0<0.001 I Nonparametric Not Significant I Analysis of Variance I [ ] Significant at p<_0.05 I Dunnett's Test Not Significant A. Kruskal-WallisTest (.5.75%tiesat artyconcentratio'_) [I I Significant at 0<--0.05 I Dunn's Test Not Significant B. Fisher's Exact Test on Proportion of Ties (>7s%tiesmtanyconeer_tration) B. Analysis of Variance with Repeated Measures I I I Significant at .o<0.05 Not Significant I ! I (Dosage) Dunnett's Test (Dosage x Block interaction) One-way ANOVA for each block I II Significant at p<0.05 I Dunnett's Test Not Significant III. Test forProD0rtion Data Variance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilities are reported as either p<0.05 or p<0.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. 418-028:PAGE 2-19 Adult data was evaluated with the individual rat as the unit measured. Litter values were used in evaluation of pup data, as appropriate. Variables with interval or ratio scales of measurement, such as body weights, feed consumption values, latency and errors per trial scores in behavioral tests and percent mortality per litter were analyzed as described under the Parametric heading of the schematic. Bartlett's Test of Homogeneity of Variances (14)was used to estimate the probability that the dosage groups have different variances. A non-significant result (p>0.001) indicated that an assumption of homogeneity of variance was not inappropriate, and the data was compared using the Analysis of Variance _15).If that test was significant (p<0.05), the groups given the test substance were compared with the control group using Dunnett's Test(16).If Bartlett's Test was significant (p_0.001), the Analysis of Variance Test was not appropriate, and the data was analyzed as described under the Nonparametric heading of the schematic. When 75% or fewer of the scores in all the groups were tied, the Kruskal-Wallis Test (iv)was used to analyze the data, and in the event of a significant result (p<0.05), Dunn's Test(18)was used to compare the groups given the test substance with the control group. When more than 75% of the scores in any dosage group were tied, Fisher's Exact Test(19)was used to compare the proportion of ties in the groups. Data from the motor activity test, with measurements recorded at intervals (Blocks) throughout each test session, were analyzed using an Analysis of Variance with Repeated Measures (2),as described under that heading in the schematic. A significant result (p<0.05) in that test could have appeared as effect of Concentration (differences among dosage groups in the totals of all measurements in a session) or as an interaction between Concentration and Block (differences in the patterns of dosage group values across the measurement periods). If the Concentration effect was significant, the totals for the control group and the groups given the test substance were compared using Dunnett's Test6). If the Concentration x Block interaction was significant, an Analysis of Variance (15)was used to evaluate the data at each measurement period, and a significant result (p<0.05) was followed by a comparison of the dosage groups using Dunnett's Test (_6). Variables that had graded or count scores, such as litter size, the number of trials to a criterion in a behavioral test or the day a developmental landmark appeared, were analyzed using the procedures described under the Nonparametric heading of the schematic. Clinical observation incidence data, as well as the descriptive and quantal data from the FOB, were analyzed as contingency tables using the Variance Test for Homogeneity of the Binomial Distribution (z_). Sperm motility data were expressed as percentages and analyzed, as indicated above, by parametric methods. 418-028:PAGE 3-1 3. RESULTS - Male Rats 3.1. Mortality, Clinical and Necropsy Observations (Summaries - Tables B1 and B2; Individual Data - Tables B17 and B18) 3.1.1. Mortality All male rats survived to scheduled sacrifice. 3.1.2. Clinical Observations All clinical observations were considered unrelated to the test substance because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in only one or two male rats in any dosage group. These observations included chromorhinorrea, missing/broken or misaligned incisors, chromodacryorrhea, lacrimation, localized alopecia on the head, underside or limbs, dry, brown perioral substance, soft or liquid feces, scab on head and dehydration. 3.1.3. Necropsy Observations All necropsy observations were considered unrelated to the test substance because the incidences were not dosage-dependent; and/or 2) the observation occurred in only one or two male rats in any dosage group. These observations included discolored/mottled kidneys, slight pelvic dilation of the left kidney, small epididymides, small testes and diverticulum jejunum. 3.1.4. Histopathology (APPENDIX J) No treatment-related microscopic changes were observed in any of the male rats administered 0.3 or 1 mg/kg/day of the test substance. Treatment-related microscopic changes were observed in the liver and thyroid gland of male rats in the 3 and 10 mg/kg/day dosage groups. The treatment-related microscopic changes in the liver consisted of minimal to moderate enlargement (hypertrophy) of centrilobular hepatocytes. The affected hepatocytes were enlarged with an increased amount of dense eosinophilic granular cytoplasm. The treatment-related microscopic changes in the thyroid gland consisted of an increased incidence of the male rats in the 3 and 10 mg/kg/day dosage groups with hypertrophy (enlargement) of follicular cells and hyperplasia (increased follicular cells and small follicles). Although the incidence in the 10 mg/kg/day dosage group was minimally increased over the control group values, the hypertrophy and hyperplasia could have been associated with the liver-cell changes. 418-028:PAGE 3-2 3.2. Terminal Body Weights and Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight and Brain Weight (Summaries - Tables B3 through B5; Individual Data - Tables B19 and B20) Terminal body weights of the male rats were slightly reduced (approximately 6%), albeit not significantly, in the 10 mg/kg/day dosage group, as compared with control _oup values. The absolute weights of the liver, the ratios of the liver weights to terminal body weights and ratios of the liver weights to brain weight were si_fificantly increased (p<0.05 and/or p<0.01) in the 3 and 10 mg/kg/day dosage groups, as compared with the control _oup value. The ratio of the heart weights to brain weight was significantly decreased (/7<0.05) in the 10 mg/kg/day dosage group, as compared with the control group value. The absolute and relative (terminal body and brain) weights of the male reproductive organs (left and right epididymes, left cauda epididymes, left and right testes, seminal vesicles with and without fluid and prostate), brain, left and right kidneys and adrenals, spleen, thymus and heart (except relative heart weight as described above) were comparable across dosage groups and did not differ significantly. 3.3. Hematology and Clinical Chemistry (Summaries - Tables B6 and BT; Individual Data - APPENDIX K) Hemoglobin concentrations (HGB) were significantly decreased (p_<0.05or p___0.01i)n the 1, 3 and 10 mg/kg/day dosage groups and average values for red blood cells (RBC) and hematocrit (HCT) were significantly decreased (p_<0.05or p_<0.01)in the 3 and 10 mg/kg/day dosage groups, as compared with the control group values. Prothrombin time (PT) was significantly increased (p<0.05 or p<0.01) in the 0.3, 3 and 10 mg/kg/day dosage groups. Dosages of the test substance as high as 10 mg/kg/day did not affect mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (PLAT), mean platelet volume (MPV), volume, white blood counts (3VBC), activated partial thromboplasfin time (APTT), nucleated red blood cell count (NRBC), lymphocytes, segmented neutrophils, bands, monocytes, eosinophils, basophils and abnormal lymphocytes in male rats. These values were comparable among the four dosage groups and did not differ significantly. Average values for cholesterol (CHOL) were significantly decreased (p<0.05 or p<0.01) in the 0.3, 1, 3 and 10 mg/kg/day dosage groups and the average values for triglycerides (TRIG) was significantly decreased (p<0.01) in the 10 mg/kg/day dosage group. Albumin (A), blood urea nitrogen (BUN), alkaline phosphatase (ALK), calcium (CA) and albumin/globulin ratio (A/G) levels were significantly increased (p<0.01) in the 10 mg/kg/day dosage group. Clinical chemistry values for total protein (TP), glucose (GLU), total bilirubin (TBILI), creatinine (CREAT), creatinine kinase (CK), alanine aminotransferase (ALT), aspartate 418-028:PAGE 3-3 aminotransferase (AST), phosphorus (PHOS), sodium (NA), potassium (K), chloride (CL) and globulin (G) in male rats were comparable among the four dosage groups and did not differ significantly. 3.4. Body Weights and Body Weight Changes (Figure 1; Summaries - Tables B8 and B9; Individual Data - Table B21) Body weight gains were significantly reduced (p<0.05 orp<0.01) in the 0.3, 3 and 10 mg/kg/day dosage groups on study days (DSs) 29 to 36. As a result of these reductions, significantly reduced (/7<0.05 or p<0.01) body weight gain occurred in the 0.3, 1, 3 and 10 mg/kg/day dosage groups on study days DS 29 to termination. Body weight gain was also significantly reduced (p<0.01) for the 10 mg/kg/day dosage group for the entire dosage period (calculated as DS 1 to termination). 3.5. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values (Summaries - Tables B10 and Bll; Individual Data - Table B22) Absolute (g/day) and relative (g/kg/day) feed consumption values for the male rats were unaffected by dosages of the test substance as high as 10 mg/kg/day. 3.6. Mating and Fertility (Summary - Table B12; Individual Data - Table B23) All mating and fertility parameters (numbers of days in cohabitation, rats that mated, fertility index, rats with confirmed mating dates during the first and second week of cohabitation and the number of pregnant rats per number of rats in cohabitation) were unaffected by dosages of the test substance as high as 10 mg/kg/day. 3.7. Functional Observational Battery (Summary - Table B13; Individual Data - Table B24) Home cage behavior of sleeping and immobile/awake were significantly different (p<0.01) in the 3 mg/kg/day dosage group, compared to the control group. These increases were not considered treatment-related because they were not dosage-dependent. There were no other statistically significant or biologically important differences among the four dosage groups in the measures of the functional observational battery (FOB). There were no alterations in autonomic functions (lacrimation, salivation, palpebral closure, prominence of the eye, pupiUary reaction to light, piloerection, respiration, defecation and urination), sensorimotor functions [responses to visual, auditory, tactile and painful stimuli (reactivity and sensitivity)], excitability (reactions to handling and behavior in the open field), gait and sensorimotor coordination (gait pattern in the open field, severity of gait abnormalities, air righting reaction and landing foot splay) and forelimb and hindlimb grip strength and abnormal clinical observations including but not limited to: convulsions, tremors, unusual behavior, hypotonia or hypertonia, emaciation, dehydration, unkempt appearance and deposits around the eyes, nose or mouth. 418-028:PAGE 3-4 Body weights recorded during the functional operational battery for the treated groups were not significantly different than the control group for the male rats. 3.8. Motor Activity (Figures 3 and 4; Summary - Table B14; Individual Data - Table B25) There were no statistically significant or biologically important differences among the five dosage groups in the number of movements or time (in seconds) spent in movement measures of motor activity on DSs 36 through 39. 3.9. Sperm (Summary - Tables B15 and B16; Individual Data - Tables B26 and B27) 3.9.1. Sperm Motility Sperm motility was unaffected by dosages of the test substance as high as 10 mg/kJday. Group mean values were comparable among the five dosage groups and ranged from 85.5% to 93.2% motile sperm. 3.9.2. Sperm Count and Sperm Density The sperm count and sperm density were comparable among the five dosage groups. No treatment-related differences were observed. 3.9.3. Sperm Morphology A low incidence of head and/or tail abnormalities was observed for the male rats in all dosage groups. No treatment-related differences were observed. 418-028 PAGE 4-1 4. RESULTS - Female Rats 4.1. Mortality, Clinical and Necropsy Observations (Summaries - Tables C1 and C2; Individual Data - Tables C28 and C29) 4.1.1. Mortality All female rats survived to scheduled sacrifice. 4.1.2. Clinical Observations All clinical observations were considered unrelated to the test substance because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in only one female rat in any dosage group except for localized alopecia which is a common finding in rats that are allowed to deliver. These observations included brown perioral substance, rales, localized alopecia on the limbs, back or head, urine-stained abdominal fur, miosis, abdominal distention, emaciation, cold to touch, swollen nose, scab or ulceration on back or head, dehydration and tip of tail missing. 4.1.3. Necropsy Observations All necropsy observations were considered unrelated to the test substance because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in only one female rat in each of the 1, 3 and 10 mg/kg/day dosage groups. These observations included the cortex of the right kidney adhered to the right lateral liver lobe in a 1 mg/kg/day dosage group female rat (19907), small left ovaries (0.003g) in one 3 mg/kg/day dosage group female rat (19051) that was not pregnant, outer capsule of spleen adhered to the omentum in another 3 mg/kg/day dosage group female rat (19063) and the large and small intestines distented with gas, small spleen and thymus, large adrenals, and approximately 12 black areas (pinpoint to 0.3 cm) on the fundic mucosal surface of the stomach in a 10 mg/kg/day dosage group female rat (19020). 4.1.4. I-lJstopathology (APPENDIX J) No treatment-related microscopic changes were observed in any of the female rats administered up to 10 mg/kg/day of the test substance. There were no treatment-related microscopic changes observed in the liver of the F1 generation pups from the dams given up to 10 mg/kg/day of the test article. There were a few microscopic changes observed in the various organs and tissues which were considered to have occurred spontaneously and not to be treatment-related. 418-028:PAGE 4-2 4.2. Terminal Body Weights, Organ Weights and Ratios (%) of Organ Weight to Terminal Body Weight and Brain Weight and Primordial Follicle Counts (Summaries - Tables C3 through C5; Individual Data - Tables C30 through C32) Terminal body weights of the female rats were comparable among dosage groups and did not differ significantly. The absolute and relative weights (to terminal body and to brain weight) of the female reproductive organs (left and right ovaries and uterus with cervix), brain, liver, left and right kidneys and adrenals, spleen, thymus and heart were comparable across dosage groups and did not differ significantly (except on one occasion). The ratio of the left kidney to the brain weight of the female rats in the 3 mg/kg/day dosage group was significantly increased (p<0.05) over the corresponding control group value. This significant finding was not considered treatment-related because: 1) the incidence was not dosage-dependent; and/or 2) the observation was a single occurrence. Average primordial follicle counts for the 10 mg/kg/day dosage group were comparable to the control group and did not differ significantly. 4.3. Hematology and Clinical Chemistry (Summaries - Tables C6 and C7; Individual Data - APPENDIX K) Dosages as high as 10 mg/kg/day did not affect any hematology or clinical chemistry values evaluated. Average values for red blood cells (RBC), white blood cells (WBC), hemoglobin concentration (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelets (PLAT), mean platelet volume (MPV), prothrombin time (PT), activated partial thromboplastin time (APTT), nucleated red blood cell count (NRBC), lymphocytes, segmented neutrophils, bands, monocytes, eosinophils, basophils, abnormal lymphocytes in female rats were comparable among the five dosage groups and did not differ significantly (except on one occasion). The concentration of WBCs (lymphocytes) for of the female rats in the 0.3 mg/kg/day dosage group was significantly increased (p<0.05) over the corresponding control group value. This significant finding was not considered treatment-related because: 1) the incidence was not dosage-dependent; and/or 2) the observation was a single occurrence. Average values for total protein (TP), albumin (A), glucose (GLU), cholesterol (CHOL), total bilirubin (TBILI), blood urea nitrogen (BUN), creatinine (CREAT), creatinine kinase (CK), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALK), calcium (CA), phosphorus (PHOS), triglycerides (TRIG), sodium (NA), potassium (K), chloride (CL), globulin (G) and albumin/globulin ratio (A/G) in female rats were comparable among the five dosage groups and did not differ significantly (except on two occasions). The concentration of G was significantly decreased (p<0.01) and the concentration of A/G was significantly increased (p<0.01) for of the female rats in the 0.3 mg/kg/day dosage group over the corresponding control group values. These significant findings were not considered treatment-related because: 418-028:PAGE 4-3 i 1) the incidences were not dosage-dependent; and/or 2) the observations were a single occurrence for both parameters. 4.4. 4.4.1. Body Weights and Body Weight Changes (Figure 2; Summaries Tables C8 through C13; Individual Data - Tables C33 through C35) Precohabitation Body weights and body weight gains were comparable and did not differ significantly during the precohabitation period at dosages of the test substance up to 10 mg/k_day. 4.4.2. Gestation Body weights and body weight gains were comparable and did not differ significantly during the gestation period at dosages of the test substance up to 10 mg/kg/day. 4.4.3. Lactation Body weight gains were not significantly affected by dosages of the test substance up to 10 mg/kg/day during lactation. Body weights were significantly reduced (/7<0.05 orp<0.01) on days of lactation (DLs) 4, 6 through 8, 11 and 13 in the 0.3 mg/kg/day dosage group, DLs 7 and 8 in the 3 mg/kg/day dosage group, DLs 4, 6 through 9, 11, 13 and 14 in the 10 mg/kg/day dosage group. These significant findings were not considered treatment-related because they were not dosage-dependent or they did not persist. 4.5. 4.5.1. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values (Summaries - Tables C14 through C19; Individual Data. Tables C36 through C38) Precohabitation Absolute (g/day) and relative (g/kg/day) feed consumption values were not significantly affected by dosages of the test substance up to 10 mg/kg/day during the precohabitation period. 4.5.2. Gestation Absolute (g/day) and relative (g/kg/day) feed consumption values were not significantly affected by dosages of the test substance up to 10 mg/kg/day during the gestation period. 4.5.3. Lactation Absolute and relative feed consumption values were not affected by dosages of the test substance up to 10 mg/kg/day during lactation. 418-028:PAGE 4-4 Absolute feed consumption values were significantly reduced (p<0.05 or p<0.01) on DLs 1 to 5 and 8 to 15 in the 0.3 mg/kg/day dosage group, DLs 1 to 5 and 8 to 15 in the 3 mg/kg/day dosage group and DLs 8 to 15 in the 10 mg/kJday dosage group. Relative feed consumption values were significantly reduced (p<0.01) on DLs 1 to 5 in the 0.3 mg/kg/day and DLs 5 to 8 in the 3 mg/kg/day dosage group. These significant findings were not considered treatment-related because they were not dosage-dependent and they did not persist. 4.6. Estrous Cycling, Mating and Fertility (Summary - Table C20; Individual Data - Table C39) The average numbers of estrous stages per 13 days were comparable among the five dosage groups and did not significantly differ. The number of rats with six or more consecutive days of diestrus or estrus did not differ significantly. All mating and fertility parameters (numbers of days in cohabitation, rats that mated, fertility "index,rats with confirmed mating dates during the first and second week of cohabitation and the number of pregnant rats per number of rats in cohabitation) were unaffected by dosages of the test substance as high as 10 mg/kg/day. 4.7. Functional Observational Battery (Summary - Table C21; Individual Data - Table C40) There were no statistically significant or biologically important differences among the five dosage groups in the measures of the functional observational battery (FOB). There were no alterations in home cage behavior, autonomic functions (lacrimation, salivation, palpebral closure, prominence of the eye, pupiUary reaction to light, piloerection, respiration, defecation and urination), sensorimotor functions [responses to visual, auditory, tactile and painful stimuli (reactivity and sensitivity)], excitability (reactions to handling and behavior in the open field), gait and sensorimotor coordination (gait pattern in the open field, severity of gait abnormalities, air righting reaction and landing foot splay) and forelimb and hindlimb grip strength and abnormal clinical observations including but not limited to: convulsions, tremors, unusual behavior, hypotonia or hypertonia, emaciation, dehydration, unkempt appearance and deposits around the eyes, nose or mouth. Body weights recorded during the functional operational battery for the treated groups were not significantly different than the control group for the female rats. 4.8. Motor Activity (Figures 5 and 6; Summary - Table C22; Individual Data - Table C41) There were no statistically significant or biologically important differences among the five dosage groups in the measures of number of movements or time (in seconds) spent in movement motor activity on DL 17. 418-028:PAGE 4-5 4.9. Natural Delivery and Litter Observations (Summary - Tables C23 and C24; Individual Data. Tables C42 through C45) Pregnancy occurred in all 15 (100%) rats assigned to the 0 (Vehicle), 0.3 and 10 mg/kg/day dosage groups and 13 (88.7%) of the rats assigned to the 1 and 3 mg/kg/day dosage groups. All pregnant dams delivered a litter of one or more liveborn pups. Values for the numbers of dams that delivered litters, the duration of gestation (calculated in days), averages for implantation sites per delivered litter, the numbers of dams with stillborn pups, the numbers of dams with no liveborn pups, dams with H1pups dying days 1 to 4 and 5 to 22, were comparable among the five dosage groups and did not significantly differ. The number of liveborn pups, the number of pups found dead or presumed cannibalized on day 1, and days 2 to 8, 9 to 15 and 16 to 22 postpartum were comparable among the five dosage groups and did not significantly differ. The gestation index (number of dams with one or more liveborn pups per number of pregnant rats), viability index (number of live pups on DL 8 per number of liveborn pups on DL 1) and lactation index (number of live pups on DL 22 per number of liveborn pups on DL 8) were compared to the control group value and did not differ significantly. The number of pups surviving per litter, pup sex ratios, litter size and pup body weights per litter on DLs 1, 8, 15 and 22 were comparable among the five dosage groups and did not significantly differ. 4.10. Pup Clinical and Necropsy Observations (Summary - Tables C25 and C26; Individual Data - Tables C46 and C47) No clinical or necropsy observations in the F1 generation pups were attributable to dosages of the test substance as high as 10 mg/kg/day because: 1) the incidences were not dosage-dependent; and 2) the observation occurred in only one to two litters. These clinical observations included: not nursing, emaciation, dehydration, whole body discolored purple, scab, cold to touch, left eye discolored purple, fight eye enlarged, laceration on right hindlimb, bruise on back, not nesting, pale in appearance and corneal opacity of right eye. Necropsy observations of 228, 203, 196, 172 and 211 pups in the five respective dosage groups on postpartum days 5 and 22 were limited to moderate dilation of the renal pelvis of one pup from a 0.3 mg/kg/day dosage group litter. 4.11. Pup Liver Weight and Ratio of Liver Weight to Terminal Body Weight (Summary - Table C27; Individual Data - Table C48) F1 generation male and female pup terminal body weights, absolute liver weight (by sex) and ratio of liver weight to terminal body weight (by sex) were comparable across all five dosage groups and did not differ significantly. 418-028:PAGE 4-6 REFERENCES 1. Sanders, J.E., Eigenberg, D.A., Bracht, L.J., Wang, W.R., and Zwieten, M.J. (1988): Thyroid and liver trophic changes in rats secondary to liver microsomal enzyme induction caused by an experimental leukotriene antagonist (L-649,923). Toxicology and Pharmacology, 95: 378-387. 2. Organisation for Economic Co-operation and Development (1996). OECD Guideline for Testing of Chemicals. Section 4, No. 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test, adopted 22 March 1996. 3. Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. 4. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. 5. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 6. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive andMutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 7. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone. (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 8. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Crl:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 9. Institute of Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 10. Haggerty, G.C. (1989). Development of Tier I neurobehavioral testing capabilities for incorporation into pivotal rodent safety assessment studies. J. Amer. Col. Toxicol. 8:53-70. 11. Irwin, S. (1968). Comprehensive observational assessment: Ia. A systemic quantitative procedure for assessing the behavioral and physiologic state of the mouse. Psychopharmacologia (Berlin) 13:222-257. 12. Moser, V.C. (1989). Screening approaches to neurotoxicity: A functional observational battery. J. Amer. Col. Toxicol. 8:85-94. 418-028:PAGE 4-7 13. O'Donoghue, J.L. (1989). Screening for neurotoxicity using a neurologically based examination and neuropathology. J. Amer. Col. Toxicol. 8:97-116. 14. Sokal, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances.Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. 15. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 16. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-1121. 17. Sokal, R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometo,, W.H. Freeman and Co., San Francisco, pp. 388-389. 18. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241-252. 19. Siegel, S. (1956). Nonparametric Statistics for the Behavioral Sciences. Fisher's Exact. McGraw-Hill Co., New York, pp. 96-105. 20. SAS Institute, Inc. (1988). Repeated measures analysis of variance. SAS/STAT TM User's Guide, Release 6.03 Edition, Cary, NC, pp. 602-609. 21. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. 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A 0 0 _ oo_oooo_oo H 0000000000 oooooo_ooo _o 0 .. _ _0 _ 0 _ _ N 0 m U 0 UO .......... o _ 0 U _0 g _ m i M 0 *. i M N H _ _0 _o _ _ eH _ om _ , _ _ O _ _ N_N_NN_NN _ 0 _ 418-028:PAGE B-94 APPENDIX C REPORT TABLES - Fo GENERATION FEMALE RATS 418-028:PAGE C-1 418-028:PAGE C-2 418-028:PAGE C-3 418-028:PAGE C-4 418-028:PAGE C-5 i o _ >o r.. ,_ o 0 Z P..l _ 0 El o E-, _ 0 _> rn r_ m, _m ,_ m o c_ o o o -Ho -Ho -_o -_o -Ho +_o +l -_ +l -H_' o o o o _ co m o - o o o o _ 0 r_ 0 0 0 0 _ tll r_ _io _o _ tll _0 0 0 0 +t +10 +10 +_O +10 "HO +10 +10 "HO -H 4"4 4"t +1_ ,_ co ,_ ,_ _ _ _, p o _ _ _ o _ o o o o .0 o _ o o o o o o o o o o o 0 I o _-- o= H o O_ X E_ 0 "" ,'r, r.v.1 _,.,, _ _ 0 0 0 0 _ _, p- ._ ._ oo" oo o_ o_ oo _o" _ I_l 0 _ 0 0 0 0 0 0 _ _ _ ..o+.o-.o-.o+,o._o..,o._o.. .. ._ ._' _" c'_ _ o o 0 0 o o 0 0 _ __ 0 HO H O_ o_ '_ o 0 o o _o m m +l .._o _o -So 440 -_o ..HO 440 "_o 4.4 +t +_ -H_ __ - o o oo o o oo _ _ ,4 d 6 _ d ,4 _ S d d d S d _Z "" o _ _ ,_ _ _ r_, r4 pe..1 [-, oo ooOO H _ _ _ = _ ,-1 ,-.1 1_ H r/l 0 _ o_ " 418-028:PAGE C-6 418-028:PAGE C-7 418-028:PAGE C-8 418-028:PAGE C-9 418-028:PAGE C-10 i 418-028:PAGE C-11 H 0 Z O > O O 121 "_ O "_ [-' >,, ,._ ,0--t -_" .__ 0 0 _:_ _0 _ _,_ . ;>1 U ,-I E E ._ ,.-I r,. 0 I_ ,-_ _ -H = Y=I >_ o _ 0_1 0 0 0 0 0 0 _ 0 _ .w ._ ,_l ,.-1 u_ i-i i 0 m r_ o m N o o o_ _'1 u 0_ " ' H 0 ro 0 _'_ i! _ - |E-_I !I !! m _:_ iI _ l___I _0_ I IHI IO_ I_IOI _ I"_I | _,._| O_1_ gxl _l_l _-_ ,_I II _} _ i_ _'_ C _ _) [ 0 _I " l 418-028:PAGE C-12 i .IN... r._ . . _o ". _ "" I o o Olo o ,--t o o 1 ! | I C) _'t 0 0 ! 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U ,--1 [.-, 418-028:PAGE C-46 418-028:PAGE C-47 H U N X 0 O O O H o _ H O O - o _____ H Q 000 _- _ _H _ 0 O_ " _ _ _ 418-028:PAGE C-48 H o _o __ 418-028:PAGE C-49 418-028:PAGE C-50 ii 418-028:PAGE C-51 418-028:PAGE C-52 418-028:PAGE C-53 418-028:PAGE C-54 418-028:PAGE C-55 u 0 _ o_ o_ _ _ o _ 0 _i 0 _ o _ .......... oooooooooo ._ _gg__ _ .......... __.._ ._ _____._ _ gggggggggg _ oooooooooo ___ _ ___ _ .......... _ oooooooooo _ ggg;ggggg_gg;g; g m _ ___og_ o _ ooooooooooooooo _ 'gggdggggggggggg j _ ____ 0 -. _o _ __ _ _ _ _ _._.._....... o _ _ o.o..o..o..o..o. oooo u _a z _ oooooooooo z_ 6666666d66 _> _ _" _ _ O_ _ n 0__ ._ u . _ ......... _ .............. _ _ 0 o o oo o _ _ _ __ 418-028:PAGE C-56 418-028:PAGE C-57 _ 0 _ _ ___ _ oooooooooo o o _ gdddgddgdg _ 44_X4_ _ _ _ ._ _ _ o _ ___ H _ OoOooodooo o _ 0 i 0 _ o_ _. _ _J_A_A_ _ __ n_ .......... oooooooooo M _ _H_ _ ,___ _ dddddgddggddg _ _ o_ ............. _____ _ oooo0oooo _ _0 _0 _ o_ _ __ O_ " m o_ mo m _ .......... _ oooooooooo _ .......... _ oooooooooo _ _ 0 0 _ _ __ oooooooooo _ _ _ __ 0 _ _ _o _ _ _ II_ _ _ _ _. _ _-_ _ __ 418-028:PAGE C-58 o> ._ $___ _ _ ___ ._ _ _ 0 0 _ o >o _ oooooooooo _ ___ _ __Z__ _ o.o..o..o..o..o..o..oooooo _ . o _ j o-. _ _N N_ _ _ __._...................... ._ Ng_N_NN_NN _ .......... _0 _ _o_ _ 0 o __ oooooooooo _ _ 0000000000000 .......... ._ NN_N_NNg_ _ oooooooooo _ __ _ _ oooooooooo __ oo "_ __ _o. _-- "_ o _0 o, _ _ ___ _0 _ ____ _ ,,_ ___ o 418-028:PAGE C-59 oo oooo o o _ m m 0 z o_ _ _ N H _ o o 0 _o _ _d _ _o_o_o_oooooo_o ggggg_gggg __ . _ _____ __ ggggggggggggggg _ __ .0 _ H 0 _ OOOOOOOOOO _ _0__ _ 4444444444 __ o...o....o.. ooodooo d _Z_ _g _ ____ _ 000000000000000 _ dgdddddddddgggd __ ____ ............... g ooooooooooooooo O _ -_ _o d __ _o __ _0 _ _0 _ _o _ O_ _ _ _o .......... __ _ _ _ _ _m_ m_ m .......... _ .......... o o'o o o o o o o o ___ _m .......... _ oooooooooo ,,_ ._ _ _ _ _ _ _ O_ -- o., _ _ 0 _ o_o_o_ _ .......... 0 _ 0 HH_H_HM_ _ _ O _ _o__oo_ _ ............... _0_._ 0 _ l, _ _ __ H_H_H_HH_HHH_ oo _II 418-028:PAGE C-60 H _ _ X _ _ o 0 _ _ o > 0 _ _ _H 0 o _ .................... 0000000000 .................... 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II el) o_, ,_,_,_ I l_il Iel ' '' _ ,o, I pc, I ['-I ,I 0_ I II 0 I C./_I I v0 _ '_'II II I 0,-,, _I_ I I _,_ __ _ _ _,_ _ 0 _D o _ _, .,_._.,_ . rl " _iI _ _ _o _,_ _o _11 I ,_i ! "_i I I |I I I i! _I __I: 418-028:PAGE C-66 418-028:PAGE C-67 418-028:PAGE C-68 H U X 0 < Q m _ 0 m _ m m _ _ 0 o >_ _ 0 ...................... _ M 0 0o i _ 0 O_ .............................. ''" ....... ''' ......... __ _ _ _ ............................ _ 0 d_ ..... 0 ' ..... _0 _ uo u_ _ __o_o_oooooooooo o o__o_o_;oooo oooooo O_ _ _,,_ H U H 0 _ o > O U _ ............... __ ___ ............... __ m o ............ 418-028:PAGE C-69 _ M O ,- O _ 0 HO _ ............... _ _ _ _O _O._.H._.O._.O._._....... , _ 0 _ __H_ 0 _ zH_o N_ O_ _ _ II_ i i i ............... U H 418-028:PAGE C-70 A d __ _ o i ................ mmmm_ X _ 0 ,. m 0 _ _ O_ _o _ o o____ .............................. ____ om_m_oo_o_ - ____ N 0 0 _ _0 ,_ H H M ............... N O O H 0 _ H .............................. 418-028:PAGE C-71 _O N_ _o M _ _ o_ .. _ o __.o_._. i o_ ............... _ ____ ooooooooooooooo _oo ,_ _ H X Z _ ................. o _ommm_moo_mm_o_ O oH 0 a g O M 0 -" _ _ _ .............................. _O H 0 H .............................. i o_ 418-028:PAGE C-72 m m m _mm_mo_mmm_m_m NN _ 418-028:PAGE C-73 418-028:PAGE C-74 H U H X ............... O _ __O_O_O_ m O 418-028:PAGE C-75 _ 0 __ H a_ _0 _ zz _ _ N o ............................... _mmm_b_mmmm_ _ _ 0 u _d O_ _ o_ _ _ ____ ____ ,,_g_ i ............... H U H 418-028:PAGE C-76 p _ 0H _ ___ _ _ 0 __o__o_ H_ _ N H 0 .. m _0 H _ N ............................... .............................. . _ _ ,_ _ _ m _ _ _ u _ __ _ _ 0_ 418-028:PAGE C-77 418-028:PAGE C-78 H U H _ X ............. 0 _ _o_ o_o_ > Q 0 _ o _o_ _o__ n0 __ 0 0 0 _ _ _ _ _ _ _ 0_ -. , 0 H -- .........._.............. o _0 _ _ _ __ _ __ .......................... o _ _ _.__o_o_o 0 _ o_ _ flflfl_flHfl_flflHflflH __ _ _ _o__o_ fl_flflHflHHHHHHHHH _ _ _ m _ ._ _ _o _ C _ __o_ _._ 0 _: _ 418-028:PAGE C-79 418-028:PAGE C-80 418-028:PAGE C-81 H U H X 0 H _ M O _, o_ 0 H H _ , _O _ o- _ _ _ _ o. =' _ _ _ _ oo __ o_ .......................... __ _o _ , . . _ _8 __ O_ _ ___ o__o__o_o_oooooooo_o_, _ H LJ 0 ,< 0 n Z 0 H i p- J _ 0 i O _o _o I-4 0 .. _ 8_ _, 0 Ho ,_ > "" o_ to o _. [.-, ,-1 _ _ _ 418-028:PAGE C-82 o-_ _ m.= _ _ ._ ,,._. 418-028:PAGE C-83 H H O 0 0 H H _ U o _, _ H 0 _ H o_ o.. _ _ _ ............... _0 . _ o_ 418-028:PAGE C-84 oo _ ._ 418-028:PAGE C-85 418-028:PAGE C-86 418-028:PAGE C-87 H X 0 O O H Z o u H O O m i H oH o0 _ 0 X _ _ m O" O _o _ _ O_ _ ................... 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O _ u N _o _ N_ _ m O_ _ .............................. _ o_ . oo _ ___=_ _ ' _ 0 _ _ _ _ o _ _ _ o =____ _ __ 8_ _0 O _ H_ ____ _= _ ooooooo_oooo_ooo 418-028:PAGE C-114 o U H O 0 _ _ _ oH 0 _ m U _ _ 0 0 m._...... ............. mmm_ m m _w _m w_ m _g _ m_ __ ........................ 0 ............. 0 ........... 0 , o _ n m. _ m m _ 0 _ N _ _ _ _ 0 _0 _ _ _ O_ _ o _ a _H oo O_ -- . _ H _ _ ................... H g ............. _ __ _ oo _ _ _ ___ _ .......................... o_ _ _ ____ _ ..... _mm_ o __ _ _ ...... _ 0 _ _ ....... ..__ _ __ ____ ? . _ _ II H 0 _ oN 0 0 0 _ 0 Z W H o _ O _ ...... _ ....... O .............. 0 , _ _ _ o _ m _ _ _ .......... _ m ............... 0 "- _ _ _o_ __o_ 418-028:PAGE C-115 N 0 _ mo _ Z H_ _ _ _ _ __ = _0 _ Z_ _ O_ _ H OH _ _ ................ _ ooo__o__ o_ ............... . _ g__ ............... _ o_ ____ ! _-__ _ _H ! _,,_ _0 m _ o H X O 0 _ _ _ o_ 0 _ g _ 0 ............... 0 , _ ............... _ 0" _ _ _Om__O0_ 418-028:PAGE C-116 . _ g 0 ............ _ ............ - _ 0 o___m_ - _ _ _0 _ _. _ O_ _ _o _ _.__o_ _ __,_oo__ _, . _ _ i_ ............... i ............... _o, __o_o___o_.o_o__,_ _ _0 0 _ _ _ _ _ _ _ 418-028:PAGE C-117 o H X O 0 _ m 0 _ " 0 _ bh 0 "- g 0 _ _ o o _ _ .... 0 ............ .... _ ............ _ _o_ _o_ _ N S a _ _ _ _ H __0 o zz _ .......................... _ o -x oo -_ .._........ _.... _ ............. _ . _ __ _ _ _ _ _ 0 _ _ _ _0 0 _ _ I H H X 0 _ o O _ g _ o .............. 0 _ o .............. o.. _O o _ m _oo_o__oo ____ ................. 418-028:PAGE C-118 _ _. _0 _ _ _ ._ m _ _0 _ _0 0 U _ _ _ o _ _ _ ____ _ ..... oooooo_ooo _ o O 0 _ H o 418-028:PAGE C-119 _ _ _ _ _ _ _ 0 " o o _ _ ............... __ooo_o_ _ ............ o _o_o _o _ _ _. _ _o _ _ _o o_ _ _ _ _ _ .............................. o_ -. ____ 0 _ _ _ __o__ _ _o _ o u H X O _ ....... D o _ ............. ; z_ 0 _ ........................ o _ _ _ _ ........................ _o _ _ 418-028:PAGE C-120 _ . _ ........... _ _000_ H _- z0 _u _ _ _ 0 D - _ _ _ _ _ m 0 _ _ _ oo O_ _ _ _ __ o o o __o__oo.ooo _ o oo _ _ 418-028 :PAGE C- 121 418-028:PAGE C-122 o H H X O N ....... 418-028:PAGE C-123 _ " oH 0 , o_ _.............. __ ._._ o_ ..... ........... ...... H __ _ _ _ _ ....................... _ _ ....................... _ ......................... ..... 0 X _7 - _ _ _ _ _ _ _. ......... _ ................. _''_ _ _0 " __ _o 418-028:PAGE C-124 418-028:PAGE C-125 418-028:PAGE C-126 418-028:PAGE C-127 418-028:PAGE C-128 418-028:PAGE C-129 418-028:PAGE C-130 418-028:PAGE C-131 418-028:PAGE C-132 418-028:PAGE C-133 418-028:PAGE C-134 418-028:PAGE C-135 418-028:PAGE C-136 H X 0 Z 0 > 0 H U 0 m _ o _= m _ .......... _ ............................. ! .......... , i ........... ! N 0 . n _ H _ -- H 0 ' __ 0_ _ _ _ o_ _ l I, I_I I_I II I_I IIII I I_ I .......... IHHff_ffff_HH I I _ I '___' _I I " I,__==__, IHI .......... i 0u U '_ _'I_I '_;_=_, _o _ __'_'__,,_,_,___, ,'', I II . ','<, IIII I_I Ikl II II iI.Ii ol I_ I ........... I '___' _i I__o_| I_I II_I I I_I ii II _,' I_ I .......... I '_____' I___I I _,, ,_ ___' I_1 .......... _I I___I I! _I_I_'___I_ IHI ........... II 'II_' ,_,_o ___'_'_g_S_=_'_ 01_.I0...,_____'_', II II I_I 'II_' II_I0 I0 00 ii ,___" II II ! m |_ _,o II , I_ I !_ _ i 418-028:PAGE C-137 H u H 0 0 Z 0 _ _i ......... i i .......... i _ _ o _ II I I _ __ o _ _ _I '.......... _ l I' .......... I , _ ___ _ _ _I' ......... _o I' .......... _ _ _0 I __ O I Ii I_1 II II_I I If ,, ! ......... II I_1 I%1 II I0! II I .......... _ __ _ _ _,_ ,___ >__I Im._.._................. ,o__=_,_o_o_o, l_m_m__ i , , g_ I_ ......... ,_, ........... _ I_ '_' ! o '_'_'___,__,,_o,o _ _ i_ ' '__ '_' _N_I O| I0 o, _,,___,_,__' APPENDIX D PROTOCOL AND AMENDMENTS 418-028:PAGE D-1 9osa,_ ore.,B_. ^ p^ 19o44 TeJepho(n2_15)443.8710 Telef_"(2/5) 443-8587 ARGUS RESEARCH CharlesPJverLaboratories Discoveryand Development Services PROTOCOL 418-028 SPONSOR'S STUDY NUMBER: 1"-7706.1 STUDY TITLE: Oral (Garage) Combined Repeated Dose Toxicity Study ofT-7706 with the Reproduction/Developmental Toxicity Screening Test PURPOSE: The purposeofthisstudyistoprovideinformatioonn thepossiblheealth hazardsthatmay resulftromrepeateedxposureofCrI:CD(SD)IGS BR VAF/PIus@ maleand femaleratstoa tesstubstancbeeginningbefore cohabitatiotnh,roughmatingandcontinuinfgoratleas4t2 days(male ratso)rthroughparturitiuonntidlay21 oflactati(ofnemalerats)T.his repeatedosestudyincorporataesreproduction/developmetnotxailcity screenintgesthatcanbe usedtoprovidienitiianlformatioonn possible effectosn male andfemalereproductipveerformance(e.g.g,onadal functiomna,tingbehaviorc,onceptiond,evelopmentoftheconceptuasnd parturitioTnh)e. studyalsoplacesemphasison neurologicaelffectassa specifiecndpointand shouldidentiftyheneurotoxipcotentiaolfa test substancweh,ich may warrantfurtheirn-deptihnvestigation. TESTING FACILITY: STUDY DIRECTOR: SPONSOR: Because of the selectivity of the endpoints and the short duration of the study, the screening test will not provide evidence for definitive claims of no reproduction/developmental effects. In particular, it offers only limited means of detecting postnatal manifestations of prenatal exposure or effects that may be induced during postnatal exposure. ArgusResearch 905 SheehyDrive,BuildingA Horsham,Pennsylvania19044-1297 Telephone: (215)443-8710 Telefax: (215)443-8587 Raymond G. York,Ph.D.,DABT Associate Director of Research F.,mail: raymond.york@criver.com Addressascited above for Testing Facility 3M CorporatTeoxicology 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 418-028:PAGE D-2 STUDY MONITOR: JohnButerthoff, Ph.D., DABT, CIH 3M CorporateToxicology 3M Medical Department Telephone: (651) 733-1962 Telefax: (651) 733-1773 Emaih jlbutenhoff@mmm.com REGULATORY CITATIONS: Protocol 418-028 Page 2 Organisationfor Economic Co-operation and Development (1996). OECD Guideline for Testing of Chemicals. Section 4, No. 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test, adopted 22 March 1996. Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. JapaneseMinistryof Health andWelfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW OrdinanceNumber21, March26, 1997. REGULATORY COMPLIANCE: This study will be conducted in compliancewith the GoodLaboratoryPractice (GLP) regulations cited above. All changes or revisions of this protocol shall be documented,signed by the Study Director and the Sponsor,datedand maintainedwith theprotocol. The Testing Facility's Quality AssuranceUnit (QAU) will audit the protocol, the raw data and the report, and will inspect critical phases of those portions of the study conducted at the Testing Facility in accordance with the Standard OperatingProcedures of the Testing Facility. The final report will include a compliance statement signed by the Study Director that ",here, port accuratelyreflects the raw dataobtained duringthe performance of the study and that all applicableGLP regulations were followed in the conduct of the study. Should significant deviationsfrom GLP regulations occur, each will be described in detail, together with how the deviation might affect the quality or integrity of the study. Shouldany portion of the studybe conductedby a subcontractor or by the Sponsor, the Study Director will ensure that a qualified Principal Investigator is identified by the facility conducting thatportionof the study. The QAU for this facility will conductcritical phase inspections and auditrespectiveresults andreports for thatstudy portionaccordingto the SOPs of that facility. Suchcriticalphase inspection reportsand reportauditswill be submittedby the facility to the PrincipalInvestigator andthe StudyDirector. The dates of theinspections andreport submissions will be incorporatedinto a QAU Statementgeneratedby that facility andprovided 418-028:PAGE D-3 l_otaco1418.028 Page3 to the Testing Facility for inclusion in the final report. In addition, this facility will provide a statement of GLP compliance, as described above, signed by the Principal Investigator for inclusion in the final report. SCItEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. TEST SUBSTANCE AND VEHICLE: Identification: Test Substance: T-7706 [Pcrfluorohexane Sulfonate Potassium Salt (PFHS)] Lot identification will be documented in the raw data. The Sponsor will provide to the Testing Facility documentation or certification of the identity, composition, method of synthesis, strength and activity/purity of the test substance. This documentation will be included in the final report. Vehicle: Aqueous 0.5% carboxymahlycellulosc (CMC) (medium viscosity) prepared using reverse osmosis membrane processed deionized water (R.O. dcionized water). Lot identification and Supplier will be documented in the raw data. Neither the Spomor nor the Study Director is aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. Therefore, no anal_es other than those mentioned in this protocol will be conducted. Safe W Precautions: Gloves, dust-mist/HEPA-filtcred mask, appropriate eye protection and uniform/lab coat to be worn during formulation preparation and dosage. The Material Safety Data Sheet (MSDS) is attached to the protocol (ATTACHMENT 2). Bulk Test Substance: Bulk Vehicle Components: Prepared Test Substance and Vehicle Formulations: Room temperature. Room t=npcraturc. Refrigerated (20C to 80C). All test substance shipments should be addressed to the attention of Julian Gulbinski, Manager of Formulation Laboratory, at the previously cited Testing Facility address and telephone number. 418-028:PAGE D-4 Protocol 418-028 Page 4 Shipments should include information concerning storage conditions and shipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment. FORMULATION: Frequency of Preparation: Formulations (suspensions) will be prepared weekly at the Testing Facility. Detailed preparation procedures will be attached to this protocol (ATTACHMENT 3). Adjustment for Purity: The test substance will be considered 100% pure for the purpose of dosage calculations. Testing Faeili_, Reserve Samples: The Testing Facility will reserve a sample of each lot of bulk test substance (approximately 1 g) and bulk vehicle components (approximately 1 g or 5 mL) used during the course of the study. Samples will be stored under the previously cited conditions. ANALYSES: Results of required analyses will be provided to the Testing Facility for inclusion in the study report. Samples additional to those described below may be taken if deerned necessary during the course of the study. Additional analyses, if required, will be dooumented by protocol amendment. Bulk Test Substance Sampling: A sample of approximately 1 g of the test substance will be taken on the last day of treatment and sent (ambient conditions) to: Principal Investigator: Lisa Clemen 3M Environmental Teelmology and Safety Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone: Telefax: (651) 778-5568 (651) 778-6176 Email: laelemenl_ztmm.eom Services The recipient will be notified in advance of sample shipment. 418-028:PAGE D-5 Protocol 418-028 Page 5 Analyses of Prepared Formulations: Concentration and Homogeneity: Concentration and homogeneity of the prepared formulations will be verified during _e course of this study. Quadruplicate samples (2 mL each) will be taken from the top, middle and bottom of each concentration on the first day of preparation. Two samples from each quadruplicate set will be shipped for analysis; the remaining samples will be retained at the Testing Facility as backup samples. Quadruplicate samples will be taken from each concentration on the last day of preparation. Two samples from each quadruplicate set will be shipped for analysis; the remaining samples will be retained as backup samples. Backup samples will be stored under the previously cited conditions and discarded at the Testing Facility upon the request of the Sponsor. StabiliW Stability of the prepared formulations will be documented during this study. Two sets of duplicate samples (2 mL each) from each concentration will be taken on the first day of preparation. One sample of each duplicate set will be shipped on the day of preparation. These samples will be analyzed at the following time points: as soon after preparation as possible and ten days after the first analysis. The remaining samples will be retained at the Testing Facility as backup samples. Backup samples will be stored under the previously cited conditions and discarded at the Testing Facility upon the request of the Sponsor. Shipplne Instructions: Samples to be analyzed will be shipped (refrigerated) to: Principal Investigator: Lisa Clemen 3M Environmental Teelmology and Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone: (651) 778-5568 Telefax: (651) 778-6176 F_.mail: laclemen@mnma.eom Safety Services The recipient will be notified in advance of sample shipment. DISPOSITION: Prepared formulations will be discarded at the Testing Facility. All remaining bulk test substance will be returned to: Dan Hakes 3M EHSR - Auto & Chern Cap 3M Center, Building 236-1B-10 St. Paul, Minnesota 55144-1000 Telephone: (651) 733-2392 418-028:PAGE D-6 TEST SYSTEM: Protocol418-028 Page6 Species/Strain and Reason for Selection: The Crl:CD(SD)IGS BR VAF/Plus rat was selected as the Test System because: 1) it is one mammalianspecieascceptedforuseintoxicitsytudieasnd ithasbeen widelyusedthroughout industr2y);thistraionfrathasbeendemonstratetdobe sensitivteoreproductivaend developmentatloxinsa;nd 3)historicdaaltaand experienceexisatttheTestingFacilictlya) Number: Initipaolpulatioancclimated: 100male and 100virginfemalerats. Populatiosnelectefdormain study: 75 male and75 virginfemalerats(15persexper dosagegroup). Populatiosnelectefdortoxicokinetsitcudy:15male and 15 femalerats(threpeersexper dosagegroup). Body Weight and A2e: Male rats will be ordered to weigh from 275 g to 300 g each at receipt, at which time they will be expected to be at least 60 days of age. Female rats will be ordered to weigh f_om 200 g to 225 g each at receipt, at which time they will be expected to be at least 56 days of age. Actual body weights will be recorded the day after receipt and will be documented in the raw data. The weight ranges will be included in the final report. At study initiation, the weight variation of the rats will not exceed _-20% of the mean weight of each sex. Sex: Both male and female rats will be evaluated. Soure_._.ee: Charles River Laboratories, Inc. The rats will be shipped in filtered cartons by air freight and/or truck from Charles River Laboratories, Inc., to the Testing Facility. Identifieafioa: Rats are permanently identified using Monel self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male and female rats are assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study before admi_stration of the first dosage. Pups will not be individually identified during lactation; all parameters will be evaluated in terms of the litter. 418-028:PAGE D-7 ANIMAL HUSBANDRY: Protoco4l 18-028 Page7 All cage sizes and housing conditions are m compliance with the Guide for the Care and Use of Laboratory Animals _a).Argus Research is an AAALAC-accredited facility. Housing: Fo generationratswill be individually housed in stainless steel wire-bottomed cages except during the cohabitation and postpartum periods. During cohabitation, each pair of rats will be housed in the male rat's cage. Beginning no later than day 20 of presumed gestation, Fo generation female rats will be individually housed in nesting boxes. Each dam and delivered litter will be housed in a common nesting box during the postpartum period. Nesting Material: Nesting material (bed-o'cobs_) will be provided. Bedding will be changed as often as necessary to keep the animals dry and clean. Analyses for possible contamination are conducted semi-annually and documented in the raw data. Room Ai,'rT, emperature and HumidiW: The animalroom is independently supplied with at least ten changes perhour of 100% fresh air that has beenpassed through99.97% HEPA filters. Roomtemperaturewill be maintainedat 640Fto 790F(18"C to 260C) and monitored constantly. Room humiditywill also be monitored constantlyand maintainedat 30% to 70%. Li__: An automaticallycontrolled 12-hour light:12-hour darkfluorescentlight cycle will be maintained. Each dark period will begin at 1900hours EST. The light cycle may be adjusted by the Study Director or designee if deemed necessary to accommodate scheduled laboratory activities. Any such adjustment will be documentedin the raw data. Die__tt: Rats will be given Certified Rodent Diet #5002 (PMI Nutrition International) available ad libitum from individual feeders. Feed will be removed the evening priorto the scheduled sacrifice. Water: Waterwill be available ad libitum from individualbottles attachedto the cages or from an automatic watering access system. All waterwill be from a local source and passed througha reverseosmosis membrane before use. Chlorine will be addedto the processedwater as a bacteriostat;processed water is expected to contain no morethan 1.2 ppm chlorine at the time of analysis. Wateris analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. 418-028:PAGE D-8 Protocol418-028 Page $ Contaminants: Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the certified diet, in the drinking water or in the nesting materials at levels that would interfewrieththeresultosfthistudy.Thereforen,o analyseostherthanthoseroutinely performedby thefeedsupplieorrthosementionedinthisprotocowlillbe conducted. DAY NUMBERING SYSTEM: Gestatiodnay 0 isdefinedasthedayspermatozoareobservedina smearof thevaginal contents and/or a copulatory plug observed in aim. The dayofbirthisdesignateldactatidoany0 (postpartudmay0)intheHealthEffectTsest Guideline-sReproductioanndFertiliEtfyfect(sOfficoefPreventionP,esticideasnd Toxic Substance8s70.3800A,ugust,1998)andintheOECD GuidelinfeortheTestingofChernical-s Combined RepeatedDose ToxicitSytudywiththeReproduction/DevelopmenTtoaxlicity ScreeninTgest(Sectio4n,No.422,22 March 1966).Thissame dayisdesignatedday I postpartu(mdayI oflactatioinn)theStandardOperatinPgrocedureosftheTestingFacility. Throughoutthisprotocolt,hedayofbirtwhillbe designatedday Ipostpartum(day I of lactatioann)d allsubsequenatgesoftheF1 generatiornatsanddaysofthelactatiopneriodwill be determineadnd citedaccordingly. RANDOMIZATION AND COHABITATION: Upon arrival, rats will be assigned to individual housing on the basis of computer-generated random units. During an acclimation period of at least five days, male and female rats will be selected for study on the basis of physical appearance and body weights recorded during acclimation. The rats will be assigned to dosage groups based on computer-generated (weightorderedr)andomizatiopnrt_edures. Within each dosage group, consecutive order will be used to assign rats to cohabitation, one male ratper female rat. The cohabitation period will consist of a maximum of 14 days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in sial will be considered to be at day 0 of presumed gestation and assigned to individual housing. Female rats not mated within the first seven days of cohabitation will be assigned alternamtaele ratsthathavemated(samedosagegroup)andwillremainincohabitatiofnora maximum ofsevenadditiondaalys. Day 1 of lactation (postpartum) is defined as the day of birth and is also the first day on which all pups in a litter are individually weighed (pup body weights will be recorded after all pups in a litter are delivered and groomed by the dam). Litters will not be culled during the lactation period, because random selection of pups for culling could result in potential biases in pup viabilities and body weight gains over this period. 418-028:PAGE D-9 Protocol 418-028 Page 9 Within each dosage group, consecutive order will be used to assign the first 10 male and the first 10 female rats to a functional observational battery (FOB) and motor activity assessment, blood sample collection for clinical chemistry and hematology (CC&H), and histological evaluations. On day 22 postpartum, a table of random units will be used to select five male and five female pups per litter for blood sample and liver collection; these pups will only be selected from the ten dams selected for FOB, motor activity, CC&H and histological evaluation. ADMINISTRATION: ,) Route and Reason for Choice: The oral (gavage) route was selected for use because: 1) in comparison with the dietary mute, the exact dosage can be accurately administered; and 2) it is one of the possible mutes for environmental exposure. Method and Frequency: Dosages will be adjusted daily for body weight changes and given at approximately the same time each day. The first day of dosage is designated as day 1 of study. Male rats will be given the test substance once daily beginning 14 days before a cohabitation period that consists of a maximum 14 days. Dosage will continue through the day before sacrifice, after completion of the cohabitation period, after a minimum of 42 days of administration. Female rats will be given the test substance once daily beginning 14 days before a cohabitation period that consists of a maximum of 14 days. Dosage will continue through the day before scheduled sacrifice (day 21 of lactation). Rationale for Dosage Selection: Dosages were selected by the Sponsor based on previous studies conducted with the test substance, taking into account possible differences in sensitivity between pregnant and nonpregnant rats. The highest dosage will be expected to cause toxic effects but not mortality or obvious suffering. The descending sequence of the lower dosage levels will be selected for the purpose of demonstrating any dosage-related response, with no adverse effects expected at the lowest level. 418-028:PAGE D-IO Protocol 418-028 Page I0 Dosage Leve_ Concentrations and Volumes: Dosage Group I II HI IV Number ofl_ts _ sex 1$ + 3b 15 + 3b 15 + 3b 15 + 3b I_ (._./k_JOay) 0 0.3 I 3 Concmm_u_n' _mlpn_L 0 0.03 0. l 0.3 Dosage Volume lncrxl) !0 I0 10 I0 ArpsBatchN.mb_ B-418-028-A(Dsy.Momh.Yesr) B-418-028-B(Day.Monoh.Year) B-418-028-.C(Day.Mm'xth.Year) B-4 ! 8-028-D(Day.Month.Year) V 15+3 b ] 10 1 l0 B-418-028-E(Day.Mont_-,.Ye)ar a. Thetestsubstancewill be considered100%purefor thep_ of dosagecalculations. b. Threeadditionalrats persex per doui_ Broupwillbe a.._g3_l m toxJcokinetic_m'_iecollection. TESTS_ ANALYSES AND MEASUREMENTS - Fo GENERATION: Viability - Male and Female Rats: All Periods: At least twice daily. Clinical Observations and/or General Appearance - Male and Female Rats: Acclimation Period: Weekly. Dosage Period: Daily before dosage. On the first day of dosage, postdosage observations will be recordedat approximately hourly intervals for the first four hours and at the end of the normal working day. Subsequent postdosage observations will be recorded at intervals deemed appropriate by the Study Director or designee after determination of peak toxicologic effects, Maternal Behavior: Days 1, 5, 8, 15 and 22 postpartum. Observed abnormal behavior recorded daily. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or Study Monitor. Data collected for rats assigned to toxicokinetic sample collection will not be summarized or analyzedstatistically. Detailed Clinical Observations - Male and Female Rats: Once before the first dosage and at least once weekly thereafter, detailed clinical observations will be conducted for all male and female rats. These observations will be made outside the cage in a standard arena at the same time each day of conduct. Effort will be made to ensure that variations in the test conditions are minimal and that observations are conducted by observers unaware of treatment groups. Signs noted should include,but not be limited to: changes in skin, fla', eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g., lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture 418-028:PAGE D-11 Protocol418-028 Page 1l and response to handling as well as the presence of clonic or tonic movements, stcrotypic behavior (e.g., excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behavior (e.g., self-mutilation, walking backwards) should also be recorded. Body Weights - Male and Female Rats: AcclimatioPneriod: Weekly. DosagePeriod: Daily. Sacrifice: Terminal weight. Feed Consumption Values - Male Rats (recorded and tabulated): DosagePeriod: Weekly. FeedConsumption Values - Female Rats (recordeadndtabulated): DosagePeriod: Weeklytocohabitation. GestatioPneriod: Days 0,7,I0,12,15,18,20 and 25 (ifnecessary). Postpartum Period: Days I,5,8 and 15. Feed consumption not tabulated after day 15postpartum, when it is expected that pups will begin to consume maternal feed. Feed Consumption Values - Male and Female Rats: Feed consumption values may be recorded more frequently than cited above if it is necessary to replenisthefeed.Duringcohabitatiownh,en two ratsoccupythesame cagewithonefeedjar, replenishment of the feed jars will be documented. Individual values will not be recorded or tabulated. Toxieoginetie Sample Collection: On day 14 and42 ofstudyb,loodsamples(approximateIlyniLeach)willbe collectefdi'om eachmaleratassignetdothetoxicokinetsiacmplecollectipoonrtionofthestudy(3pm group). On day 14 ofstudyandday21 ofpresumedgestatiobnl,oodsamples(approximatelIymL each) will be collected from each female rat assigned to the toxicokinetic sample collection portion of the study (3 per group). Samples will be collected prior to dosage on day 14 of study. The time of each blood collection will be recorded in the raw data. 418-028:PAGE D-12 Protocol418-028 Page 12 Blood will be collected from the orbital sinus. If necessary, blood may be collo:ted from an alternate site; if so, the alternate site will be documented in the raw data.) The samples will be transferred into EDTA-coated (purple top) tubes and spun in a centrifuge. The resulting serum will be transferred into polypropylene tubes labeled with the protocol number, Sponsor study number, animal number, sex, group number, dosage level, day of study, coUection interval, date of collection, species, generation and storage conditions. All samples will be immediately frozen on dryiceandmaintainedfrozen(<-70C)untilshipmentforanalysis. ARer thelasbtloodsamplecollectiorna,tswillbe sacrificaenddsamplesoftheliverwillbe collectefdoranalysis. Shipping Instructions: Samples to be analyzed will be shipped on dry ice to: Principal Investigator: Lisa Clemen 3M Environmental Technology and Safety Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone: (651) 778-5568 Telefax: (651) 778-6176 Email: laclemen@hnmm.com Services The recipient will be notified in advance of sample shipment. Estrous Cycling and Matin2: Est_us cycling will be evaluated by examination of vaginal cytology beginning with the day after the first administration and then until spermatozoa are observed in a smear of the vaginal contents and/or a copulatory plug is observed in aitu during the cohabitation period. Caesarean-Sectioning - Toxicokineti Study: On day 21 of presumed gestation, blood and liver samples will be collected from all female rats designated for toxicokinetic sample collection. Blood samples will be collected from the rats as previously described. After sacrifice, the liver of each rat will be excised and the liver weight will be recorded. The median liver lobe will be frozen and stored (_<-20C) until shipment for poss_le analysis. The fetuses will be removed from the uterus and blood samples will be collected from each fetus via decapitation. Blood will be placed into tubes, pooled per litter, allowed to clot and spun in a centrifuge; the resulting serum will be transferred into labeled polypropylene tubes. All samples will be immediately frozen on dry ice and maintained frozen (_<-70C) until shipment for analysis. 418-028:PAGE D-13 Protocol41 S-028 Page 13 Thelivefrromeachfetuwsillbecollectpeodo,ledperliRearndplacedintolabeletdubesT.he samplewsillbefrozeanndstore(d_<-20uCn)tislhipmenftoranalysis. SampleswillbeshippedondryicetoLisaClcmenatthepreviouscliyteadddress. Natural Delivery: Female rats will be evaluated for: Adverse Clinical Signs Observed During Parturition. Duration of Gestation (day 0 of presumed gestation to the time the first pup is observed). Litter Size (defined as all pups delivered). Pup Viability at Birth. Functional Observational Battery: Onone occasion during the course of the study, a functionalobservationalbattery (FOB)(5-s)will beconductedon 10 male and 10 female ratsper group. Formale rats,this assessmentwill be conductedshortlybefore scheduled sacrifice,but priorto blood sample collection for hen__atologaynd clinical chemistry evaluations. Femaleratsshould be tested during the lactation period,shortly before scheduled sacrifice. The FOB, to be conducted by an observerunawareof the groupassignmentof the rat, will assess the following parameters: 1. Lacrimation,salivation, palpebralclosure,prominence of the eye, pupillary reaction to fight,piloerection,respiration,andurinationand defecation (autonomic functions). 2. Sensorimotorresponsesto visual, auditory,tactileand painful stimuli (reactivity and sensitivity). 3. Reactions to handlingand behaviorin the open field (excitability). 4. Gait patternin the open field, severity of gait abnormalities,airfighting reaction, and landing foot splay(gait andsensorimotorcoordination). 5. Forelimb and hindlimbgrip strength. 6. Abnormal clinical signs includingbut not limited to convulsions, tremorsand other unusualbehavior,hypotoniaor hypertonia,emaciation, dehydration, unkemptappearanceanddepositsaroundthe eyes, nose ormouth. 418-028:PAGE D-14 Protocol 41S-O2g Page 14 Evidence of the ability of this battery to detect the effects of positive control substances will be provided (Testing Facility Positive Control Data). Data will also be provided to document interobserver reliability if more than one observer is involved in the testing. Motor Activity Test: Motor activity will be evaluated on 10 male and 10 female rats per group once during the course of the study. This assessment will be conducted shortly before scheduled sacrifice, but prior to blood sample collection. The movements of each rat will be monitored by a passive infrared sensor mounted outside a stainless-steel wire-bottomed cage (40.6 x 25.4 x I7.8 cm). Each test session will be 1.5 hours in duration with the number of movements and time spent in movement tabulated at each fiveminuteintervalT.he apparatuwsillmonitora rackofup to32 cagesandsensorsduringeach sessionw,itheachrattesteidnthesame locatioonn therackacrosstestsessionsG.roupswillbe counterbalancaecdrostsestinsgessionasnd cages. Datawillbe providedtodemonstrattehatthetesstystemiscapableofdetectinigncreaseisn activiptryoducedby positivceontroslubstance(sTestinFgacilitPyositivCeontrolData). HEMATOLOGY AND CLINICAL CHEMISTRY: At scheduled sacrifice, the rats (fasted) assigned to hematology and clinical chemistry (H&CC) sample collection will be exsanguinated fi'om the inferior vena cava following sacrifice by carbon dioxide asphyxiation. Approximately 5 mL of blood will be collected and processed as described below. Deternfinations additional to those described below may be conducted tfthe known properties of the test substance may, or are suspected to, affect related metabolic profiles (e.g., calcium, phosphate, fasting triglycerides and fasting glucose, specific hormones, and cholinesterase). Hematoloev: Approximately I mL of blood will be collected into EDTA-coated tubes and maintained on wet ice or refrigerated until shipment for analysis of the following hematologic parameters: Erythrocyte Count (RBC) Hematocrit (HCT) Hamoglobin (HGB) Mean Corpuscular Hemoglobin Mean Corpuscular Hemoglobin Concentration (MCHC) (MCH) Mean Corpuscular Volume (MCV) Leukocyte Count, Total (WBC) Leukocyte_ Differential Platelet Count (PLAT) Mean Platelet Volume (MPV) Cell Morphology Two blood smear slides will be prepared at the Testing Facility for each sample for measurements of differential leukocyte count. All samples (on wet ice) and slides (ambient conditions) will be shipped to Redfield Laboratories at the following address. Approximately 1.8 mL of blood will be added to a tube containing 0.2 mL of soditun citrate (0.129 lVl). The contents will be mixed and maintained on wet ice until the tubes are centrifuged 418-028:PAGE D-15 Protoco4l 18-028 Page15 (within 30 minutes of the collection time). The resulting plasma will be transferred 2.0 mL polypmpylene tubes labeled with study number, Sponsor's study number, rat number, dosage level, day of study, collection interval, date of coUcction, species, generation and storage. All sampleswill be frozen on dryice and maintained frozen (__70C) until shipment on dryice by overnightcourierfor measurement ofprothrombin time (PT) and activatedpartial thromboplastintime (APTT). Clinical Chemistry: Approximately2 mL of blood will be collected into serum separatortubes and centrifuged. The resultingsera samples will be immediately frozen on dryice andmaintained frozen (_<-70C) until shipment for analysis of the following parameters: TotalProtein (TP) Triglycerides(TRI) Albm'nin(A) Globulin (G) Albumin/GlobulinRatio (A/G) Glucose (GLU) Cholesterol(CHOL) Total Bilixubin(TBILI) UreaNitrogen (BUN) Creatinine(CREAT) CreatinineKinsse (CK) Alanine Aminotransferase(ALT) AspartateAminotransferase(AST) Alkaline Phosphatase(ALK) Calcium (CA) Phosphorus (PHOS) Sodium (NA) Potassium(K) Chloride (CL) Samples will be shipped (on dryice) to RedfieldLaboratoriesatthe following address. Shippln_ Instructions: Samples will be shipped to arriveon MondaythroughFriday accordingto the conditions describedabove to: Principal Investigator: Ms. Phyllis Powell Redfield Laboratories A Division of CRL-DDS 100 East Boone Street P.O. Box 308 Redfield, Arkansas 72132 Telephone: Telefax: (501) 397-2540 (501) 397-2002 Therecipient will be notified in advance of sample shipment. URINALYSIS: Urinalysiswill not be conducted unless indicated basedon expected or observedtoxicity of the test substance. 418-028:PAGE D-16 METHOD OF SACRIFICE: Protocol418-028 Page 16 Fo generatiornatswillbe sacrificbeyd carbondioxideasphyxiation. GROSS NECROPSY AND HISTOPATHOLOGY -Fo GENERATION RATS: ScheduledSacrific-eToxicokinetiSctudy: Schedulesdacrifiocfemaleratswillbe conducteodn day42 ofstudy.Scheduledsacrifiocfe femaleratswillbe conductedon day 21 ofpresumedgestation. Bloodsampleswillbe collectefdromtheratsaspreviousldyescribedA.ftersacrificteh,eliver ofeachratwillbe excisedandthefiverweightwillbe recordedT.he medianliverlobewillbe fi'ozcanndstored(_<-20Cu)ntilshipmentforanalysisF.etalsampleswillbe collecteads previousldyescribed. Carcassewsillbe discardewdithoutfurtheervaluation. Sampleswillbe shippedon dryicetoLisaClemenatthepreviouslcyitedaddress. Scheduled Sacrifice - Main Study: Scheduled sacrifice of male rats will be conducted on the day following the last dosage administratiaofnt,eraminimum of42 daysofdosage.Scheduledsacrifiocfefemaleratswill be conducteodn day 22 oflactation. Grossnecropsyofall male and femaleratswillincludaen initial physicaelxaminatioonf externaslurfaceasnd allorificeass,wellasthecraniatlh,oraciacnd abdominalcavitieasnd their contents. Special attention will be paid to the organs of the reproductive system. The number of implantation sites and corporalutea will be recorded. Male and female rats will be examined for gross lesions. Gross lesions will be retained in neutral buffered 10% formalin and examined histologically. Tissue trimming and histopathology will be performed under the supervision of or by a Board-Certified Veterinary Pathologist. The ovaries and the uterus with cervix of each female rat will be weighed, and ovaries, uterus, vagina and a _ gland will be retained in neulral buffered 10% formalin. Uteri of apparently nonpregnant rats will be examined after being pressed between glass plates to confirm the absence of implantation sites, and retained in neutral buffered 10% formalin. 418-028:PAGE D-17 Protocol 418-028 Page 17 Sperm Evaluations of Male Rats: To assess the potential toxicity of the test article on the male reproductive system, the endpoints listed below will be evaluated from the first I0 male rats in each dosage group. Organ Weights:The followinogrganswillbe individualwleyighed:rightestisl,eft testilse,Repididymi(swholeand cauda)r,ighetpididymiss,cn-,invaelsicle(swithand withoutfluida)nd prostate. Sperm EvaluationsS:perm concentratiaonndmotilitwyillbc evaluateudsingcomputerassistesdpermanalysi(sCASA). Motilitwyillbe evaluatebdy theHamiltonTheme IVOS by collectioofna samplefi'omthelefvtasdefcrensA. homogenatewillbe preparedfromthelefctaudaepididymifsorevaluatiboyn theHamiltonTheme IVOS to determinsepermconcentrati(osnpermpergram oftissuweeight).The remainingportion oftbelefctaudaepididymiwsillbe usedtomanuallyevaluatsepermmorphology. Sperm morphologyevaluationwsillincludethefollowingl:)determinatiofnthe percentagoefnormalspermina sampleofatleas2t00;and2)qualitatievvealuatioonf abnormalsperm,includinsguchcategorieasabnormalhead,abnormaltaila,nd abnormalhead andtail. SecATTACHMENT 4 foradditiontailssuetsobe weighedandretainefdromthetenratspersex pergroupassignemd histologicsaalmplecollectiaonndevaluation. Allothertissuewsillbe discarded. Scheduled Sacrifice of Female Rats that Do Not Deliver Litters: Rats that do not deliver a litter will be sacrificed on day 2S of presumed gestation. Gross necropsy, examination and tissue retention will be conducted as described previously for rats at schedulesdacrifice. Dams withNo SurvivingPups: Dams withno survivinpgupswillbe sacrificaefdtetrhelasptup isfounddead,missingor presumedcannibalizedG.rossnecropsye,xaminatioanndtissureetentiownillbe conductedas describeadboveforratsatscheduledsacrifice. 418-028:PAGE D-18 Rats Found Dead or Moribund: Protoco4l18-028 Page18 Rats that die or are sacrificedbecauseof moribundcondition, abortionor premature delivery will be examined for the cause of death or moribund condition on the day the observation is made. The rats will be examined for gross lesions. Testes and _ididymides of male rats will be excised and paired organ weights will be recorded. The epididymides will be rvtained in neutral buffered 10% formalin. The testes will be fixed in Bouin's solution for 48 to 96 hours and then retained in neutral b_ff_,,d 10% formalin. Pregnancy stares and uterine contents of female rats will be recorded. Aborted fetuses and/or delivered pups will be examined to the extent possible. Uteri of apparently nonpmgnant rats will be examined after being pressed between glass plates to confirm the absence of implantation sites. Ovaries and uteri will be retained in neutral buff_-d 10% formalin. TESTS_ ANALYSES AND M_ASUREMENTS - F1 GENERATION: Viability: Preweaning Period: Litters will be observed for dead pups at least twice daily. The pups in each fitter will be counted once daffy. Clinical Observations and/or General Appearance: Pmweaning Period: Once daily. Pups will be observed if they are warm and clean, for evidence of a nest and if pups are grouped together and nursing or have milk in stomach. Each pup will be examined for general shape of the head, trunk, limbs, tail and presence of anus. Clinical observations may be rvcorded more frequently than cited above, if deemed appmpriatv by the Study Director and/or the Study Monitor. Body Weights: Pmwcaning Period: Days 1 (birth), 8, 15 and 22 postpartum. Sacrifice: Terminal weight. Feed Consumption Values (recorded and tabulated): Preweaning Period: Not recorded. METHOD OF SACRIFICE - FI GENERATION PUPS: FI generation pups will be sacrificed by carbon dioxide asphyxiation. 418-028:PAGE D-19 Protocol41g-028 Page 19 NECROPSY - FI GENERATION: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation. Unless specifically cited below, all other tissues will be discarded. Pups Found Dead on Day 1 Postpartum: Pups that die before examination of the litter for pup viability will be evaluated for vital status at birth. The lungs will be removed and immersed in water. Pups with lungs that sink will be identified as stillborn; pups with lungs that float will be identified as liveborn, and to have died shortly after birth. Pups with gross lesions will be preserved in Bouin's solution for possible future evaluation. Pups Found Dead or Moribund,, on Days 2 to 4 Postpartum: Pups found dead or sacrificed because of moribundity will be examined for gross lesions and for the cause of death or the moribund condition. Pups with gross lesions will be preserved in Bouin's solution for possible future evaluation. Scheduled Saerifiee: On day 22 postpartum, pups will be will be sacrificed and examined for gross lesions; gross lesions will be preserved in neutral buffered 10% formalin. Necropsy will include a single crosssection of the head at the level of the frontal-parietal suture and examination of the crosssectionebdrainforapparenhtydrocephaly. Blood samples will be collected fxom each selected pup (5 per sex per litter from the 10 females per group selected for FOB and motor activity assessment, blood sample collection for H&CC, and histological evaluations) from the veua cava. The blood will be placed into tubes, pooled per littearl,lowedtoclotandspunina centrifugteh;eresultinsgerumwillbe transferriendtolabeled polypropylenteubes.Allsampleswillbe immediatelfyrozenon dryiceandmaintainefdrozen (_<-70Cu)ntislhipmentforanalysisT.he livefri'omeachselectepdup willbe collecteedxcised andtheorganweightrecordedT.he medianlobewillbe fi'ozeanndstored(_<-20Cu)ntil shipmentforpossiblaenalysisF.rozensampleswillbe shippedtoLisaClemen atthepreviously citedaddressT.he remaininpgortionofeachlivewrillbe retaineidnneutrabluffere1d0% formalifnorpossiblheistologiceavlaluationT.he liverwsillbe processeadndevaluated histologicaalsldyescribefdortheFo generatiornatsinHistologicEavlaluatioinn ATTACHMENT 4. 418-028:PAGE D-20 i PROPOSED STATISTICAL TESTSO']6): The following schematic represents statistical analyses of the data. Protocol 418-028 Page 20 I. Parametric TVDe of Test a II. Nonpararnetrhic A- BarUelfs Test = I I I Significant at/__0,001 Not Signlllcant I I Nonpararnatrtc Analysisof Varlance I [ I Significant at ;o<0.05 I Not SIgnnlcant A. Kruskal-WallisTest (! 7_%tm at=ay=rceftraeon) I I I Signilicantat p<_0.05 I Dunn'a Test Not Signif_ant B. Fisher'zExact Test on ProportionofTies (=.TSk'J=t- _ =nc:a._n_n) B. Analysis of Variance wie= Repeated Measures [ f J Significantit p__o.o5 Not 8ignir_ant ! (Dosage) Dunnatfs Test I I (Do_le x Blo_ Intmtce_) One-way ANOVA for each block I II Significant at p._:O.0_5 Not Significant I III. Test for Proportion Data Vadance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilities are reported as eitherp < 0.05 orp < 0.01. b. Proportion data are not included in this category. e. Test for homogeneity of variance. 418-028:PAGE D-21 Protocol 418-028 Page 21 Test items in the FOB using interval scales, such as the grip-strengthtests and the landing foot splay test, as well as body weight data and feed consumption values will be analyzed as described under the Parametric heading of the schematic. Bartlett'sTest of Homogeneity of Variances9)will be used to estimate the probabilitythatthe groupshad differentvariances. A nonsignificant result (p>0.001) will indicate that an assumption of homogeneity of variance is not inappropriate,and the data will be comparedusing the Analysis of Variance Testc1).If that test is significant (p_<0.05),the groups exposed to the test article/substance will be compared with the control group using Dunnett's Test(1). If'Bartlett'sTest is significant (p-0.001),the Analysis of Variance Test is not appropriate, and the data will be analyzed as described under the Nonparametricheading of the schematic. When 75%or fewer of the scores in all the groups are tied, the Kn_kal-Wallis Test02)will be used to analyze the data, and in the _,nt of a significant res,uit (p_<0.05),Dunn'sTest03)will be used to compare the groups exposed to the test article/substance with the controlgroup. When more than 75%of the scoresin any group are tied, Fisher'sExact Test04)will be used to comparethe proportion of ties in the groups. Data fio, m the motor activity test, with repeatedmeasurementswithin a session, will be analyzed using an Analysis of Variance with RepeatedMeasures0_3,as describedunder that heading in the schematic. A significant effect (p_<0.05)in that test can appear as effect of Concentration(a differencebetween groups in the total acrossall measurements in a session) or as an interaction between Concentration and Block (a differencebetween groups at specific measurement periods). If the Concentration effect is significant, the totals for the control group and the groups given the test article/substancewill be compared using Dmmett's Test. If the Concentrationx Block interactionis significant, an Analysis of Variance Test will be used to evaluate the data at each measurementperiod, and a significant result(p_<0.05)will be foUowedby a comparison of the groups using Dunnett'sTest. Test items in the FOB having graded or count scores will be analyzed using the procedures described under the Nonparametric heading of the schematic. Clinical observation incidence data, as well as the descriptive and quantal data from the FOB, will be analyzed as contingency tables using the Variance Test for Homogeneity of the Binomial Distribution. Alternate or additional statistical evaluations may be performed if deemed necessary or appropriate. 418-028:PAGE D-22 Protocol418-028 Page22 DATA ACQUISITION_ VERIFICATION AND STORAGE: Data generated during the course of this study will be recorded either by hand or using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, the Coulbourn Instruments Passive lnfrared Motor Activity System, the Coulbourn Instruments Auditory Startle System, the Coulbourn Instruments Spatial Delayed Alternation System, and/or the passive avoidance software. All data will be tabulated, summarized and/or statistic.ally analyzed using the Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microaofl Excel [part of Microsoft Office 97 (version SR-2)] and/or The SAS System (version 6.12). Records will be reviewed by the Study Director and/or appropriate management personnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the dra_ final report, after which time the Sponsor will be contacted to determine the disposition of these materials. KEY PERSONNEL: Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Director of Research: Alan M. Hoberman, Ph.D., DABT Associate Director of Research and Study Director: Raymond G. York, Ph.D., DABT Director of Operations and Compliance: Barbara J. Patterson, B.A. Director of Laboratory Operations: John F. Barnett, B.S. Director of Study Management: Valerie A. Sharper, M.S. Manager of Animal Operations: Dena C. Lebo, V.M.D. Chairperson, Institutional Animal Care and Use Committee: Douglas B. Learn, Ph.D. Consultant, Veterinary Pathology: W. Ray Brown, D.V.M., Ph.D., ACVP 418-028:PAGE D-23 RECORDS TO BE MAINTAINED: Protocol and Amendments. Test Substance, Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (if prescribed by Staff Veterinarian). General Comments. Clinical Observations and/or General Appearance. Body Weights. Feed Consumption Values. Natural Delivery Observations. FOB and Motor Activity Observations. Blood Sample Collection, Processing and Shipment. Gross Necropsy Observations. Organ Weights. Photographs (if required). Study Maintenance (room and environmental records). Feed and Water Analyses. Packing and/or Shipment Lists. FINAL REPORT: Protocol418-028 Page23 The Study Director will provide periodic updates of study progress to the Sponsor. Draft summary tables ofunaudited computer-recorded data may accompany these updates. Statistical analyses will not be performed on these interim data. A comprehensive draft final report will be prepared on completion of the study and will be finalized following consultation with the Sponsor. The report will include the following: Sunnnary and Conclusion. Experimental Design and Method. Evaluation of Test Results. Appendices: Figures, Summary and Individual Tables Summarizing the Above Data, Protocol and Associated Amendments and Deviations, Study Director's GLP Compliance Statement, Reports of Supporting Data (if appropriate) and QAU Statement. The Sponsor will receive one copy of the draft report and two copies of the final report. Data will be hand- and/or computer-recorded. Records will be reviewed by the Study Director and/or appropriate management personnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft final report, after which time the Sponsor will be contacted to determine the disposition of these materials. 418-028:PAGE D-24 INSTITUTIONAL Protocol418-028 Page 24 ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described in this protocol have been reviewed by the Testing Facility's Institutional Animal Care and Use Committee. All procedures described in this protocol that involve study animals will be conducted in a manner to avoid or minimize discomfort, distress or pain to the animals. The Sponsor's siguature below documents the fact that information concerning the necessity for conducting this study and the fact that this is not an unnecessarily duplicative study may be obtained from the Sponsor. No alternative (in vitro) procedures were available for meeting the stated purposes of the study. REFERENCES: 1. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive andMutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 2. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 3. I.aug, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River CrI:CD_BR //at. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 4. Institute of Laboratory Aaximal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Haggerty, G.C. (1989). Development of Tier I neurobehavioral testing capabilities for incorporation into pivotal rodent safety assessment studies. J. Amer. Col. Toxieol. 8:5370. 6. Irwin, S. (1968). Comprehensive observational assessment: Ia. A systemic quantitative procedure for assessing the behavioral and physiologic state of the mouse. Psychopharmacologia (Berlin) 13:222-257. 7. Moser, V.C. (1989). Screening approaches to neurotoxicity: A functional observational battery. J. Amer. Col. Toxieol. 8:85-94. 8. O_3onoghue, J.L. (1989). Screening for neurotoxicity using a neurologically based examination and neuropathology, J. Amer. Col. Toxicol. 8:97-116. 9. Sokal, R.IL and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. 418-028:PAGE D-25 Protocol418-028 Page 25 10. Snedecor, G.W. and Coehran, W.G. (1967). Analysis of Variance. StatisticalMethods, 6th Edition, Iowa State University Press, Ames, pp, 258-275. 11. Dunnett, C.W. (I 955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-i 121. 12. Sokal, R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 13. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometries 6(3):241-252. 14. Siegel, S. (1956). Nonparametric Statistics for the Behavioral Sciences, McGraw-Hill, New York, pp. 96-104. 15. SAS Institute, Inc. (1988). Repeated measures analysis of variance. SAS/STAT TM User's Guide, Release 6.03 Edition, Cary, NC, pp. 602-609. 16. Snedecor, G.W. and Coehran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 240-241. PROTOCOL APPROVAL: FOR THE TESTING FACILITY (_,... t.- _._fr..__ AlanM. Hoberman,Ph.D., DABT Directorof Research Ra T Study Director _.c (7,5 - _ _ TMheemftb_earH,I.nWstiotoudtimon'da,lDA.n'vi_mda.l Care and Use Committee FOR THE SPONSOR John Butenhoff,Ph.D., DABT, CIH StudyMonitorand Sponsor's Representative 418-028:PAGED-26 Protocol 418-028 Page 26 -".k_, r.. _. o "xDate Date _a._._/'4..-.,-_ a Date Date 418-028:PAGE D-27 ATTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE 418-028:PAQE D-28 A'FrACHMENT1 STUDY SCHEMATIC Protocol 418-028 rage 1of 3 COMBINED REPEAT DOSE AND REPRODUCTIVE/DEVELOPMENTAL SCREEN i TOXICITY FirstDay of Test Substance LastDay of Test Subs_.ncc Dosage Prematmg Male Rats Dosag_ Cohabitation Last Day of Test Substance Dosage" 2 Weeks 2 Weeks Motor Activity/FOBb Natural Deliv_y Prmaned Postpartum Period Rats Female __ Day _ Dry 22* / Motc_ Activity/FOBd _ Dosage Period a. For additional details see "Tests, Analyses and Measurements" section of the protocol. b. FOB and motor activity evaluations conducted on ten males per group. . Male rats sacrificed _ completion of at least 42 days of dosage; necropsy and retention of male reproductive organs. Hematology, clinical biochemistry and histological samples collected from ten male rats per group. d. Ten female rats per group assigned to FOB evaluation and motor activity evaluation. e. Ten female rats per group and their fitters sacrificed on day 22 postpartum; necropsy and retention of female reproductive organs Hemato]ogy, clinical biochemistry and histological samples collected. Remaining female rats sacrificed on day 22 postpartum and discarded. 418-028:PAGE D-29 ATTACHMENT1 Protoc4o1l8-028 Pase 2 of 3 SCHEDULE n 26 MAR 02 01 APR 02 01 APR 02 - 12 JUN 02 14 APR 02 PM - 21 APR 02 AM 21 APR 02 PM - 28 APR 02 AM 14 APR 02 15 APR 02 28 APR 02 06 MAY 02 19 MAY 02 06 MAY 02 - 07 MAY 02 Animal Receipt-AcclimatioBnegins. StartofDosagePeriod-Male Rats(14daysbefore cohabitatioandthrougha 14-daycohabitation period until the day before sacrifice after at least 42 days of dosage). Dosage Period - Female Rats (14 days before cohabitation through Day 22 of lactation). Cohabitation Period (Maximum of 14 days), Male 1 (7 days) Male 2 (7 days) Day 14 ToxicokinetiScampleCollection First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. First Possible Day 21 of Presumed Gestation Toxicokinetic Sample Collection and Sacrifice Toxicokinetic Study Female Rats Last Possible Day 21 of Presumed Gestation Toxicokinetic Sample Collection and Sacrifice Toxicokinetic Study Female Rats FOB and Motor Activity Evaluation - 10 Male Rats per Group a. The'start date of the study is the day the Study Director signs the protocol. b. Throughout this schedule, the day of birth is designated day 1 postpartum (day 1 of lactatioann)d allsubsequenatgesoftheF1 generatiornatsanddaysofthelactation periodwillbe determineadndcitedaccordinglya,sdescribeadbovetheprotocoslection, "Day Numbering System." 418-028:PAGE D-30 ATTACHM.ENT 1 06 MAY 02 23 MAY 02 l0 MAY 02 23 MAY 02 12 MAY 02 13 MAY 02 23 MAY 02 - 09 JUN 02 27 MAY 02 - 13 JUN 02 24 SEP 02 Protocol 418-O28 Page 3 of 3 FirsPtossiblDeelivery(Day21 ofpresumed gestation). LastPossiblDeelivery(.Day 25 ofpresumed gestation). FirsPtossiblDeay 25 ofPresumedGestation Female Sacrifice. LastPossiblDeay 25 of PresumedGestation Female Sacrifice. Day 42 Toxicokinetic SampleCollectioannd ScheduledSacrific-eToxicokinetSitcudyMale Rats Scheduled Sacrifice - Main Study Male Rats (Earliest possible date). Hematology, Clinical Biochemistry and Histological Sample Collection of Selected Male Rats. FOB and Motor Activity Evaluation - l0 Female Ra_ per Group. Day 22 Postpartum - Sacrifice Female Rats and Pups. Hematology, Clinical Chemistry and Histological Sample Collection of Selected Female RatsandPups. Drai_FinalReport 418-028:PAGE D-31 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 418-028:PAGE D-32 HATERIAL SAFETY OATA SHEET (Experimental} 3M 3M Center St. Paul, Hinnesota 55144-1000 1-800-354-3577 or (651) 737-6501 {24 hours) Copyright, 1999j HAnnesota Mining and Hanufacturing Company, AAJ. rZghts reserved. Copying and/or aownj.oadzng oi' thZs nforaaton ?or the purpose of properly utJ.lj.zing 3H products As aLloNed provided that: 1) the AnforaatAon As copied J.n full Nj.th no changes unless prAor agreelent is obtaAned froa 3R, and 2) neither the copy nor the orignal As resold or otherwse distributed Nth the ntentj.on o1' earnng a profit thereon. DZVISZON: 3H SPECIALTY HATERXALS MATERIAL: L-9051 DEVELOPNENTALPRODUCT ISSUED: Deceaber 07, 1999 SUPERSEDES:Hay 17, 1999 DOCL_ENT: 04-5470-2 e e e e I . e .... eeel eeee 1. ZNGREDZENT el le_ee_._elellele.e.e--em.e # aaeell....e_._._, C.A.S. NO. e - ...... PERCENT POTASSZUNPERFLUOROHEXANE_JLFONATE..... RESIDUAL ORGANZCFLUOROCHEHIGALS........ 3871-99-6 HAxture 100,0 Unknown Thj.s materAal J.s not 1j.sted for research and developaent supervj.sAon of a technj.cally on the TSCA invmntory and should be used purposes only under the dArect qualAfied ndAvAdual. ................................................. 2. PHYSZCALDATA ............................................................................. .... ........................ 8OILZNG POZNT: ................. VAPOR PRESSURE:................ VAPOR OENSXTY:................. EVAPORATIONRATE:.............. SOLUBZLZTY IN MATER:........... SPECZFXC GRAVITY: .............. PERCENTVOLATILE: .............. pH: ............................ VZSCOSITY: ..................... HELTZNG POZNT:................. NIA NeglgAble NIA Neg1.1.gAbZe slAght ca. 1.0 Hater=l (Bulk) NegligAbZe NIA N/D N/D APPEARANCEAND ODOR: Off-NhAte crystallAne solAd, sharp odor. -------------................................................................ Abbreviatj.ons: N/D - Not Determined N/A - Not Applicable CA - Approx_Jnately 418-028:PAGE D-33 MSDS: L-9051 DEVELOPMENTALPRODUCT December 07, 1999 ....... .. .................... .---t ....... .... 3. ......... FIRE AND EXPLOSION HAZARDBATA ........................ ...... t ....... ....... . ..... . ...... .. ...... . ..................... PAGE 2 . .... .. t. PLASH VU_Ni: ................... FL/_MABLE LIMrTS - LEL: ........ FL_LE LIMITS - UEL: ........ AUTOIC_ITION TEHPERA13JRE.:..... _ _1_ P _etafiash SetafZash N/A N/A N/D Closed Cup EXTINGUISHING MEDIA: Water, Carbon dioxide, Dry chelicaZ, Foam SPECIAL FIRE FZC_TI_3 PR_EDURES: Hear full protective cZoth_ung, including helmetp self-contained, positive pressure or pressure demand breathng apparatus, bunker and pants, bands around armss Meier and legs, face mask, and protective covering for exposed areas of the head. coat t_ FIRE _J_DEXPL_I_ _: See Hazardous Decomposition section for products of combustion. 4. REACTIVITY DATA STABILITY: &'table INGOHPATZBILII_f - MATER/U.S/GONDZTIONSTO AV_ID: None known. FU_ZARDOUPSOLYMERIT.ATI_: Hazardous polymerization will not occur. H_DOUS DECO_LuOSITZ_ PR_TS: Hay produce fZuorooarbon (over 300 G). gases if exposed to very high temperatures _=i.eme_.eemme._eeeW.elleeee_.eeteemeteeeelee.eele_eelle= 5. ENVIRONHENTAL INFOR_L_TZON .................... ...............o ......... .. ......... .... m_.eeete_..e.m.= .............._...... SPILL RESPONSE: Observe precautions from other sections. CoZZect spJ.Lled materiaZ. Use wet sweeping compound or water to avoid dusting. residue. Place in a closed container. Clean up REGO_E_)ED DISPO_L: Incinerate in an ndustrial or COmlercial facility tn the presence of a ombustble material. Combustion products NLll lnclude HF. OlsposaZ alternative: Dispose of waste product in a faoilty permitted to accept chemical waste. Abbreviations: N/D - Not Dete_ined NIA - Not Applicable CA - ApproxJuleteZy 418-028:PAGE D-34 MSDS: L-9051DEVELOPHENTAL Oecember 07, 1999 PRODUCT PAGE 3 ................... . ................ ... ...... . ...... . ...... . ............ ..... 5. ENVIRONMENTAL INFORHATION (continued) ............................. ................... . ........................... _NVIRONMENIAL OAIA: Not determtned. REGULATORYINFORMATION: Volatile Organic Compounds: N/D. V0C Less H20 & Exempt Solvents: N/D. Since reguZations before disposal. ErA Hazardous). vary, consuZt appZicable reguZetions or authorities U.S. ErA Hazardous Haste Number = None (Not U.$. OTHER ENVIRONMENTAL INFORHATION: This product may contain one or more organic fluorochemicals that have the potentaZ to resist environment. degradation and persist in the EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: No 6. SUGGESTED FIRST AID EYE CONTACT: ImmediateZy flush medical attention. eyes wth large amounts of Hater. Get Immediate SKIN CONTACT: Flush skin with large amounts of Hater. If *rrita*on persists, get medical attention. INHALATION: If signslsymptoms occur, remove person to fresh air. If signs/symptoms continue, call a physio_n. IF SWALLOHED: If swallmded, calZ a physician the instruction of a physician. unconscious person. _mediately. Only indues vomiting st Never give anything by mouth to an e..w ......... le_oeeeae.l_eeeel.ee_l.m_.e_.e..eele_ll_el.ele_._.e_.em..eeleOl 7. PRECAUTIONARY INFORHATION ............. . ..... . .... . ................... . ....... ......................... EYE PROTECTION: Avoid eye contact. Avoid eye contact with vapor, sprays The following should be Horn alone or _n coebination, as to prevent eye contact: Hear vented goggles. or mist. appropriate, .... .... ...... Abbreviations: ....................................... N/O "- Not Determned .... NIA - Not Applicable ........ .... .... CA - Approximately 418-028:PAGE D-35 MSDS: Lo9051DEVELOPHENTAL PRODUCT OecemDer 07, 1999 PAGE 4 ......... ..... ........... ..-..._...... ....... . .... . ...... ...... .......... .... 7. PRECAUTIONARYINFORHATION .... --- ...... ------- .................................. (continued) -.... .... o. ....... .... SKiN PROTECTION: Avoid skin contact. Near appropriate gloves Hhen handling this material. A pair of gloves made from the following material(e) are recommended: butyl rubber, polyethylenelpolyvinyidene chloride (Saranex). Use one or more of the folloNing 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 matera2s: polyethylenelpolyvinylidene chloride (Saranex). RECOHHENDEDVENTILATION: Use with appropriate local exhaust ventLtatlon. Provide appropriate local exhaust ventClatlon at transfer points. Use kn a Hell- vent1ated area. Provide sufficient ventilaton to maintain emissions below recommended exposure l_lits. f exhaust vent1aton is not adequate, use appropriate respiratory protection. Provide ventilation adequate to control vapor concentrations beloN recommended exposure lJJmits and/or control spray or mist. Local exhaust vent_Iation Bee airborne. is recommended Hhere the material beco RESPIRATORY PROTECTZON: Avoid breathing of arborne material. Select one of the foJ.lowing NIOSH approved respirators based on airborne concentration of contamCnants and n accordance Nth OSHA regulations: half-mask supp1ed aiJ" resprator, full-face suppled air respirator. PREVENTION OF ACCIDENTAL ZNGE_rZON: Do not eat, drink or smoke when using this product. Hash exposed areas thoroughly Nith soap and Hater. Hash hands after handlng and before eating. REGOmENOED STORAGE: Store at room temperature. closed when not in use. Keep container dry. Keep container FIRE ANO EXPLOSION AVOIDANCE: Not determined. EXPOSURELIMITS INGREDIENT .................................................. POTASSIUM PERFLUOROHEXANE SULFONATE.......................... RESIDUAL ORGANIC FLUOROCHEMICAL.S.... VALUE UNT ...... 0.1 HGIH3 0.1 HGIH3 TYPE ..................... AUTH SKIN* TI_ 3H V TNA 3M Y * SKIN NOTATION: L_sted substances inO_cated Hith 'Y' under SKIN refer to Abbreviations: NIO - Not Determined NIA - Not Applicable CA . ApproxJJuateZy  i 418-028:PAGE D-36 HSDS: L-9051 DEVELOPHENTALPRODUCT DecemDer 07, 1999 PAGE S INGREDIENT EXPOSURELIHITS (continue_) VALUE UNrT TYPE AUTH SKIN* the potential oontrbutiu, tu the uverall exposure by the iutaneou_ route including mucous membrane and eye, either by airborne or, more particularly, by direct contact Hith the substance. Vehiles can aZter skin absorption. SOURCEOF EXPOSURELIMIT DATA: - 3M: 3H Recommended Exposure GuideZines ..........._..................--------------......-.. 8. HEALTH HAZARD DATA ............................ EYE CONTAGT: No information _as found regarding Sngle exposure may cause: effects from eye contact. H_d Eye Irritation: sgnslsymptoms can include redness, sNelling, pain, and tearng. SKIN CONTAGT: NO lnfomatlon Nas found regarding effects from skin contact. May be absorbed through the akin and persist extended time. in the body for an Single exposure may cause: Moderate Skin Irritation: sgns/syaptoms can ncZude redness, sNelZng, itching, and dryness. INHALATION: No nforaaton Nas found regarding effects from _nhalaton exposure. Hay be absorbed by inhalation t_e. and persist _n the body for an extended Single overexposure, above recommended guidelines, may cause: Irritation (upper respiratory): sgnslsyaptoms can nolude soreness of the nose and throat, coughing and sneezing. IF SWALLOWED: Ingestion is not a likely route of exposure to this product. No information Nas found regarding effects from sNallowtng. Animal studies conducted on organic fZuorochemicaZs Nhich may be present in this product indicate effects including lvsr disturbances, weight loss, loss of appetite_ lethargy, and neuroZogicel, pancreatic, adrena and hematologic effects. There are no knoHn human heaZth Abbrevations: NIO - Not Determined NIA - Not Applicable CA - Approximately 418-028:PAGE D-37 MSDS: L-9051DEVELOPHENTAL PRODUCT OecemDer 07, 1999 .......... 8. ......... m.......... HEALTH HAZARDDATA ..... .... .... . .............................. (continued) PAGE 6 effects from antzcipated exposure to these organzc fluorochemicals when used as intended and instructed. OTHERHEALTH HAZARD INFORHATZON: This product may contain one or more organic fluorochnicals that have the potential to be absorbed and remain in the body for long periods of tJJwe, either as the parent molecule or as metabolites, and may accumulate health effects fluorochemicals w2th repeated exposures. There are no known human from anticipated exposure to these organic Hhen used as intended and instructed. The presence of organio fluor_cheskcaZs _n the population and subpopulations, such as Norkers, datLng back to the 1970's. 3M's epidNiological workers indicates no adverse effects. blood of the general has been published study of its o_n SECTION CHANGEDATES ........ . ...... .......... ....... .--- ..... ....,................................ PRECAUTIONARYINFO. SECTION CHANGEDSINCE Hay 17, 1999 ISSUE Abbreviations: N/D - Not Determined ............................................................................. NIA - Not Applicable CA - Approx_lately The lnforeation in this HateriaZ Safety Data Sheet (MSOS) is believed to be correct as of the date issued. 3H HAKESNO NARRANTZES,EXPRESSEDOR ZMPLZED, INCLUDING, BUT NOT LIHZTEDTO, N_YIRPLZEDHARPJ_WYOF MERGHANTARILITY OR FITNESS FOR A PARTICULAR PURPOSEOR COURSEOF PERFORMANCEOR USAGEOF TRADE. User is responsible for determining whether the 3R 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, soee of which are uniquely within the user's knm_ledge and control, it is essentLal that the user evaluate the 3H product to determine _hethsr it is fit for a particular purpose and sutable for user's method of use or appZication. 3H provides nformation in electronic form as a service to its custoMrs. Due to the remote possib_ity that electronic transfer Bay have resulted in errors, omissions or alterations in this information, 3R Bakes no representations as to its completeness or accuracy. In addition, information obtained from a database say not be as current as the information in the HSOS available directly from 3H. 418-028:PAGE D-38 ATTACHMENT 3 TEST SUBSTANCE PREPARATION PROCEDURE 418-028:PAGE D-39 ATTACHMEN3T Protoco4l18-028 Version4:18-028(g1 MAR02} Page1of 2 TEST SUBSTANCE PREPARATION PROCEDURE Test Substance: T-7706 Vehicle: Aqueous0.5% CMC (mediumviscosity) A. Purpose: The purposeof thisprocedureis to providea methodfor the preparationof dosagesuspensionsof T-7706 for oral(gavage)administrationto rats on Argus Research Study number418-028. B. General Information: 1. All suspensioncontainerswill be labeled and color-coded.Each label will specifythe protocolnumber,test substanceidentificationA, rgusbatch number, concentrationd, osagelevel,preparationdate, expirationdate and storageconditions. 2. Suspensionswillbe prepared: m Daily _ Weekly _ For_days of use _ Approximatelyeveryten days m By Sponsor 3. Suspensionswill be administeredat a finaldosagevolumeof 1_..m.OL0/kg. 4. Safety: Gloves,uniform/labcoat,gogglesor safetyglasseswith side shields X Dust-rnist/HEPA-filteredMask Half-Face Respiratorif notused in a chemicalfume hood w Full-FaceRespirator/PositivPeressureHood Tyvek Suit or tyvekapronand sleeves 5. Dosage suspensionsadjustedfor % Activity/Purityor CorrectionFactor:. Yes m % Activity _ No (Calculationsbased on 100%) _ % Purity m CorrectionFactor 6. SamplingrequirementsC: itedin protocol 7. Storage:Cited in protocol 418-028:PAGE D-40 " ", ATTACHMENT 3 Protocol 418-028 Version: 418-O28(19 MAR 02) TEST SUBSTANCE PREPARATION PROCEDURE Page 2 of 2 NOTE: Prior to test substance preparationaccuratelymeasurethe required amount of the appropriatevehicle (R.O. deionizedwater shouldbe used for calibrationpurposes)in a graduatedcylinder,pourthe required amount of vehicle intoa beaker. Carefully mark each beaker at the meniscus. This mark will be used duringthe preparationto bringthe test substanceslurryup to volume. C. DosageSuspensionPreparation: 1. Weigh the requiredamountof test substanceon a piece of weigh paper or intoan appropriatelysized mortar(see PREPARATION CALCULATIONS). 2. If weigh paper is used,transferthe test substanceto an appropriately sized mortar. If necessary,gdndthe test substanceintoa fine powder. Slowlyadd a smallamountof vehicle and grind.Continueto add vehicle slowlyand grindthe vehicleand the test substancetogetherto form a fine slurry. Transferthe vehicle/testsubstanceslurryto a marked beaker. 3. Rinse the mortarand pestlewithadditionalvehicleto remove any remainingtest substance. Transfer rinseto beaker. 4. Add additionalvehicleto the beaker to bringvolumeup to the mark. Place on magnetic stir plateand agitate priorto and duringsampling,aliquotting and/or administration. 5. Repeat steps (1) through(4) for each concentration. .. Written By: },:_,,/_", .,,,'t ._,,_t_r,,,z_ .,,, Clarification__N2--(JJYes [see attachedclarificationform] IniUal/Date C (L "7/3 I)0_3 ,/ 418-028:PAGE D-41 ATTACHMENT 4 TISSUES TO BE WEIGHED, RETAINED AND EXAMINED HISTOLOGICALLY 418-028:PAGE D-42 ATTACHMENT 4 Protocol 418-028 Page 1 of 2 TISSUES TO WEIGHED AND RETAINED FOR POSSIBLE EXAMINATION FROM TEN RATS PER SEX PER GROUP The tenratspersexpergroupassignetdofunctionoablservationbaaltteraynd motoractivity testwsillbe assignetdohematologyc,linicbailochemistrayndhistologiceavlaluations. Tissues to be Weiehed: The followinogrganswillbe excisedt,rimmedandindividualwleyighedassoonaspossible afteerxcisiotnoavoiddrying. liver kidneys adrenals spleen brain heart thymus ovaries testes uterus (with cervix) right epididymis prostate left epididymis (whole and cauda) seminal vesicle(swith and without fluid) Tissues to be Retained: The following tissues or representative samples will be retained in neutral buffered 10% formalin. brain (representative regions including cerebrum, cerebellum, pous) small and large intestines (including Peymas patches) lungs (perfused with neutral buffered 10% formalin) lymph nodes (submandiblar and mediastinal) peripheral nerve (sciatic or tibial) gross lesions stomach spinal cord (cervical, thoracic andlumbar) liver kidneys adrenals spleen heart thymus trachea urinarybladder testes* . thyroid/perathyroid uterus bone marrow (sternum) ovaries prostate seminal vesicles (with coagulating gland) uterus vagina mammary gland (female rats only) * Testes will be fixed in Bouin's solution for 48 to 96 hours before being retained in neutral buffered 10% formalin. Additionally, the .remaining portion of the left epididymi(scorpus andcaput), as well as the right cpididymis will be fixed in neutrabluffere1d0% formalin. 418-028 :PAGE D-43 ATTACHMENT4 Protocol418-028 Page 2 of 2 Histological Examination: Histological examination of retained tissues, including reproductive organs, will be coxJducted for the assigned ten rats per sex from the control and high dosage groups and from the F1 generation pups (livers) from the control and high dosage groups. If lesions attributed to the test substance are observed in the rats exposed to the high test substance concentration, the same tissues will be examined from the assigned ten rats per sex exposed to the lower test substance concentrations. Should re.suits warrant examination of the lower dosage groups and conduct of quantitative evaluation, scheduled report date and prices will be adjusted accordingly. The postlactional ovary should contain primordial and growing follicles as well as the large corpora lutea of lactation. Histopathological examination may detect qualitative depletion of the primordial follicle population. A quantitative evaluation of primordial follicles will be conducted for Fo generation feanale rats; the number of rats, ovarian section selection and section sample size will be statistically appropriate for the evaluation procedure used. Examination will include enumeration of the number of primordial follicles, which can be combined with small growing follicles, for comparison of ovaries of rats assigned to treated and control groups. Shippin_ Instructions: Tissues to be examinedhistologically will be shipped (ambient conditions) to: Principal Investigator: W. Ray Brown, D.V.M., Ph.D., ACVP Veterinary Pathologist Research Pathology Services, Inc. 438 E. Butler Avenue New Britain, Pennsylvania 18901 Telephone: Telefax: (215) 345 -7070 (215) 345-4326 Email: WRBRPS@concentric.net The recipient will be notified in advance of sample shipment. ' # 9os _ o_ _eg.A HorshamP, A 19044 Telephone:(215) 443-8710 r_c z/s) _3.asa7 418-028:PAGE D-44 ARGUS RESEARCH ChaHesRiverLaboratories Discoveryand DevelopmenSt ervices PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706. l Amendment 1 - 17 April 2002 1. Detailed Clinical Observations - Male and Female Rats (page 10 of the protocol): [Effective Date: I April 2002] Detailed ciinioal observations will not be recorded for male and female rats assigned to toxicokinetic sampling. Reason for Change: This change was made because sufficient data for evaluation of detailed clinical observations wig be available from the rats assigned to the main study. 2. Feed Consumption Values - Male Rats and FemaleRats (page 11 of the protocol): [Effective Date: 1 April 2002] Feed consumption values will not be recorded or tabulated for male and female rats assigned to toxicokinetic sampling. Reason for Change: This change was made because sufficient data for evaluation of feed consumption will be available from the rats assigned to the main study. Any revisions made to this finalized amendment must be made by subsequent amendment. 41B-02B:PAQED-45 3. Estrous Cycling and Mating (page 12 oft.he protocol): Protocel418-028 Amendment1 Page2 [Effective Date: 1 April 2002] For the rats assigned to toxicokinetic sampling, estroucsyclinwgillbe evaluatedduringthecohabitatipoenrioduntislpermatozoa areobservedina smearofthevaginaclontentasnd/ora copulatorpylugis observedinaim,butnotduringthedosageperiodp,riortocohabitation. ReasonforChange: Thischangewas made becausesufficiednattaforevaluationfestroucsycling willbe availablferomtheratsassignedtothemain study. 4. ScheduledSacrific-eTox.icokineSttiucdy(page16 oftheprotocol): [EffectiDvaete: 5 April2002] Thenumber ofimplantatiosniteasndcorpora luteawillbe recordedC.arcassewsillbe discardewdithoutfurtheervaluation. Reasonof Chan_e: Thischangewas made inordertoprovidemore informatioanboutpossible toxicity of the test substance in pregnant rats. 5. ScheduledSacrific-eMain Study(page16 ofthepmtoc,ol): [EffectiDvaete:27 March 2002] The number ofimplantatiosnitewsillbe recordedr,athetrhanthenumber ofimplantatiosniteasndcorporaluteawillbe recorded. Reasonof Change: The number of corporalutewaillnotbe recordedbecausecorporalutearegresast arapidrateandarenotcountedon studieastweaning. 6. ScheduledSacrific(epage19 oftheprotocol): [EffectiDvaete:4 April2002] The liveri_omeachselectepdupwillbe excised and the organ weight recorded, rather than the liver from each selected pup will be collected excised and the organ weight recorded. Any revisions made to this finalized amendment must be made by subsequent amendment. 418-028:PAGE D-46 ' Reason for Change: Protocol418-028 Amendment1 Page3 This change clarifies the protocol by removing an extraneous word. "/'-I? _anM.Hob=-,P==h,.DD._, T Director of Research -x"-- _/...0..,4::It _, '-/-/7-o&_ DateR.a_"'"'aY:orhk,ekgD..,,_T Date Associate Director oT'_esearch Study Director Thcrcsa H. Woodard, D.VMI. Member, Institutional Animal Care and Use Comm/ttee Date John Butenhofl_ Ph.D., DABT, CIH Date Study Monitor and Sponsor's Representative Any rev_lons made to this finalized amendment must be made by subsequent amendment. 418-02S:PAGE D-47 9os sr_ one, _. A Ho_l_mn.PA 19044 re_honC,:_2S_,_437.8m rdermc_:ZlSi,_3-8S87 ARGUS RESEARCH CharlesRiverL_boratories Discoveryand DevelopmentServ/ces PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706WITH THE REPRODUCTION/DEVELOPIV[ENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 Amendment 2 - 15 July 2002 1. Safety Precautions (page 3 of the protocol and Attachment 3 page 1 of the protocol): [Effective Date: 29 April 2002] A half face respirator will be worn in addition to gloves, appropriate eye protection and a uniform/lab coat during formulation preparation of the bulk test substance. Reason for Change: This change was made to match the bulk test substance text with the text located within the Material Safety Data Sheet. 2. Study Schedule (Attachment 1 page 2 of the protocol): [Effective Date: 29 April 2002] The dates for FOB and motor activity evaluations have been extended to four days (06 MAY 02 - 09 MAY 02) rather than two days (06 MAY 02 - 07 MAY 02). Any revisions to this f'malized amendment must be made by subsequent amendment. 418-028:PAGE D-48 Protocol 418-028 Amendment2 Page 2 Reason for Change: This change was made because more time is needed to evaluate animals assigned to FOB and motor activity evaluations. t obcrman, Ph.D., DABT Date Ra_iond G. York_.Ph.Df, DABT Date Study Director _L_,..L,.._-___L_...L.._..... -_,_-Thcr_ H. Woodard, D._.M. Member, Institutional Animal Care and Use Committee q.... Date I John Butenhoff, Ph.D., Study Monitor and Sponsor's Repr_entative DABT, CIH Date Any revisions to this fmallzed amendment must be made by subsequent amendment. ' 418-028 :PAGE D-49 9os_/o,_ a_, Ho=h_Pm^,Z9044 Telephone(2: 15) 443-8710 T_e_: (ZlS)_3-8S87 ARGUS RESEARCH CharlesP_er Laboratories Discoveryand DevelopmentServices PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706,1 Amendment 3 - 17 July 2002 1. Bulk Test Substance Sampling, Shippimz Instructions, Shipping Instructions, Caesarean-Sectioning - Toxicokinetic Study, Scheduled Sacrifice - Toxicokinetic and Scheduled Sacrifice, (pages 4, 5, 12, 13, 16 and 19, respectively of the protocol) [Effective Date: 29 May 2002] Samples for perfluorohexanesulfonate (PFI-IS) analysis shipped to Lisa Clemen at 3M Environmental Technology and Safety Services will be reshipped and remaining samples, that were to be shipped to Lisa Clemen and have not yet been shipped, will be shipped to: John Haherty, Ph.D. (Principal Investigator) Exygen Research 3058 Research Drive State College, Pennsylvania 16801 Telephone: (814) 272-1039, ext. 122 Telefax: (814) 231-1580 Email: john.flaherty@exygen.com Any revisions to this finalized amendment must be made by subsequent amendment. 418-028:PAGE D-50 Protocol 418-028 Amendment 3 Page 2 These samples include: Bulk test substance sampling (page 4 &the protocol) Concentration and homogeneity (page 5 of the protocol) Stability (page 5 of the protocol) Plasma toxicokinetie samples (page 11 to 12 and page 16 of the protocol) Plasma samples, rather than serum samples, were retained. Liver toxicokinetic samples (pages 12 and 16 of the protocol) Pooled fetal serum toxicokinetie samples (pages 12 and 16 of the protocol) Pooled fetal liver toxicokinetic samples (pages 13 and 16 oft he protocol) Pooled pup serum toxicokinetic samples (page 19 of the protocol) Pup liver toxicokinetic samples (page 19 of the protocol) The analyses will be subcontracted to Exygcn by the Sponsor and the Quality Assurance Unit for Exygen Research will conduct critical phase inspections and audit the respective results and reports according to the Standard Operating procedures of that facility. Such critical phase inspection reports and audit reports will be submitted by that facility to the Study Director, Raymond G. York. The date of the inspections and report submissions will be incorporated into a QAU statement generated by Exygcn Research for inclusion in the final report for Protocol 418-028, Reason for Change: This change was made at the request of the Sponsor because 3M is not able to complete the formulation analysis with their current staffing. A / Director of Research Date __1_2"_- o =_" hq,, --f_ "_'7"-'--/'7-_ Assotffate Director ogRes.R_/arch Study Director .....+,.... ,_ TMhember, Institutional An.Vim.Mal. Care Date and Use Committee .SJSothundyBMutoennihtoofrf, anPdh.D., DABT, CIH Date Sponsor's Representative Any revisions to this finalized amendment must be made by subsequent amendment. 418-028:PAGE D-51 9_ _ _ _ A .0,_ r^ _ roca_ tIis) 4#_er ARGUS RESEARCH C_.harlPesdverlaboratories _ aad DereloOment Semces PROTOCOL 418-028 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 Amendment 4- 20 March 2003 1. Bulk Test Substance Sampling, Shippin_ Instructions, Shipping Instructions, Caesarean-Sectioning - Toxicokinetic Study, Scheduled Sacrifice - Toxieokinetie Study, and Scheduled Sacrifice, (pages 4, 5, 12, 13, 16 and 19, respectively of the protocol) and Amendment 3, Item 1: Bulk Test Substance Sampling, Shiopin_ Instructions, Shivvin_ Instructions, Caesarean-Sectioning - Toxicokinetic Study, Scheduled Sacrifice - Toxicokinetic Study, and Scheduled Sacrifice (page 1 of Amendment 3) [Effective Date: 27 February 2003] The analyses performed by Exygen Research will be done according to Exygen Method ExM-023-071 Revision 1, entitled "Method of Analysis for the Determination of Perfluorohexanesulfonate (PFHS), Perfluorooetanesulfonate (PFOS) and Pentadecafluorooctanoic Acid 0aFOA) in Rat Liver, Serum and Urine Revision 1". Any revisions to this f'malized amendment must be made by subsequent amendment. 418-028:PAGE D-52 Protocol 418-028 Amendment 4 Page 2 Reason for Change: This change was made at the request of the Exygen Research in order to clarify the method of analysis. ..................................... O__A J_ Alan M. _oberman, Ph.D., DABT Date R4t_.oonnd G: Y L__+e -_-_. .D,. DABT Date Directo_ Research ............... Dbfa ag_. Learn, Ph.D. Chair Institutional Animal and Use Committee _'_'_te Care Assocmte Dtrecto_RDs'earch John Butenhoff, Ph.D., DABT, Study Monitor and Sponsor's Representative CIH Date Any revisions to this f'malized amendment must be made by subsequent amendment. APPENDIX E DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 418-028: PAGE E-1 DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 1. The following rats were not fasted overnight prior to necropsy. Dosage Group I Dosage (moJkg/day) 0 II 0.3 IT/ 1 IV 3 V 10 Date of Necropsy 28 MAY 02 29 MAY 02 28 MAY 02 29 MAY 02 28 MAY 02 29 MAY 02 28 MAY 02 29 MAY 02 28 MAY 02 Sex Female Female Female Female Female Female Female Female Female Rat Numbers 19044 19012, 19021, 19068 19009, 19016, 19036, 19043, 19047, 19052, 19055, 19061, 19071 19018, 19037 19017 19013, 19029, 19060 19045, 19058, 19073 19039, 19063, 19069 19001, 19028, 19031, 19033, 19049, 19059, 19070 This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter and there were no indications of toxicity in clinical chemistry parameters for the female rats. 2. Postdosage observations were performed out of the range of 60 _+10 minutes on the following dates. Dosage Group I IV Dosage (mg/kg/day) 0 3 Date 09 APR 02 03 JUN 02 03 JUN 02 Numberof Rats Maie Rats Female Rats 4 - 1 - 1 Range of Time Deviated (minutes) + 1to +2 +8 +7 These deviations did not adversely affect the outcome or interpretation of the study because the extent of the deviation was less than 10 minutes. 3. On day 39 of study (DS 39) (9 May 2002), a postdosage clinical observation was not recorded for one male rat in the 0.3 mg/kg/day dosage group (19153). This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 4. On DS 44 (14 May 2002), dosage volume was not recorded for one male rat in the 3 mg/kg/day dosage group (19156). This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 418-028: PAGE E-2 5. On DS 46 (16 May 2002), postdosage clinical observations and dosage volumes were not recorded for two male rats in the 0 mgJk_day dosage =,group(19161 and 19167). These deviations did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 6. Detailed clinical observations were not recorded weekly for the following rats. Dosage Group I II III IV V Dosage (mg/kg/day) 0 0.3 1 3 i0 Date 09 MAY 02 09 MAY 02 09 MAY 02 09 MAY 02 09 MAY 02 Sex Female Female Female Female Female Rat Numbers 19023, 19041. 19050 19004 19008 19054, 19062 19020, 19030 These deviations did not adversely affect the outcome or interpretation because sufficient data were available to evaluate this parameter. of the study 7. On DS 13 (13 April 2002), feed left values were not recorded for all rats before they were placed into cohabitation. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 8. On days 1 and 2 of lactation (DLs 1 and 2) (10 May 2002 and 11 May 2002, respectively), one pup in the 0 mg/g/day dosage group (litter 19050) was recorded as having an adverse clinical observation without the sex being recorded. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 9. On DL 3 (12 May 2002), one litter in the 10 mg/kg/day dosage group (19022) was not recorded as having been observed. All pups appeared normal on DL 4. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 10. On DL 5 (12 May 2002), maternal observation was not recorded for one rat in the 0.3 mg/kg/day dosage group (19018). Maternal behavior on DL 8 was normal for this rat. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 11. On 19 May 2002, the frozen liver sample for one female rat with a non-confh-med date of mating in the 0 mg/kg/day dosage group (19077) was lost before sample shipment. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. 418-028: PAGE E-3 12. On DL 22 (31 May 2002) the liver weight of one pup in the 1 mg/kg/day dosage group (pup 9 in litter 19024) was not recorded. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were available to evaluate this parameter. All deviations are documented in the raw data. __________ __3Zd'_u__.___z+_____t Ray-'yt_ndG. York,/Ph.D., _)ABT Date Associate Director o_ch Study Director APPENDIX F CERTIFICATE OF ANALYSIS 418-028:PAGE F-1 I Certificate of Analysis Nominal Product: Potassium pedluorohexanesulfonate" Product Code: PFHS; 127498-80 April 15, 2002 Torn Kestner and Joel Miller C6F,3SO3c) K(+) The sample of 127498-80 PFHS was analyzed using LCMS, 19F-NMR and ZH-NMR analysis techniques. The overall qualitative and quantitative compositional results that were derived from these combined analyses are summarized below in TABLE-1. TABLE-1 Sample: 127498-80: PFHS OverallQuantitatiCvoempositionaRlesultsby LCMS, '_F-NMR, andIH-NMR Analyses Component Identities * Relative Wt.% Concentrations (single trial measurements, excluding any water) CF3CF2CF2CF2CF2CF2SO3(') K (+) 88.93% (NMR) (CF3)2=CF-(CF2)3=SO31")K (+) 6.83% (NMR) CF3CF2-CF(CF3)-CF2CF2-SO3 (')K (+) 2.75% (NMR) CF3CF2CF2CF2CF(CF3)-SO3 (') K(+) 0.83% (NlvlR) CF3CF2CF2CF(CF3)=CF2-SO(3-K) (+) 0.56% (NM:R) (CF3h-C-CF2-CF2SO3 (') KC+) 0.078% (NMR) C6Fi2HSO3 (')K (+) 0.009% (LCMS) CTFIsSO3('K) (+) C4F9S03 (') K (*) Probable C_H2,_2 hydrocarbons Other components Sum of Cc,FI_O_ t') K t+_Isomers Sum of Known Impurities Traceamountsof otherunassignedcomponentswerealso detected. 0.006%(LCMS) 0.004% (LCMS) 0.0034% (NMR) <0.001%(LCMS) 99.98% 0.021% Page 1 of I FileReference:CofA_PFHS_127498-80.doc 418-028:PAGE F-2 3M SPECIALTY MATERIALS MANUFACTURING DIVISION ANALYTICAL LABORATORY To_.__. From: Dan Hakes - (3-2392) - EHSR - Auto & Chem Group - 236-1B-10 Tom Kestner - (3-5633) - SMMD Analytical Lab - 236-2B-11 Request. No. GID:32537 Date: Chemical Characterization April 15, 2002 of PFHS (127498-80) by 1H-NMR & 19F-NIVIRSpectroscopy SAMPLE DESCRIPTION: 127498-80: PFHS made by George Moore and to be used for toxicological Nominal product --C6FI3-SO3 (') K (+) (white powder). OBJECTIVE: testing. This sample was subjected to _H-NlVIRand _9F-NMR spectral analyses to determine the purity of the nominal product and to characterize as many impurity components as possible. Joel Miller also performed an LC/MS analysis and his results were reportedto you previously. EXPERIMENTAL: A portion of the sample (--400 mg) was totally dissolved in deuterated dimethylsulfoxide (DMSO-d_) and then the solution was _iked with a small amount of 1,4-bis(trifluommethyl)benzene (p-HFX) for NMR analysis. A 400 MHz H-NMR spectrum (# h32537.GID.401) and a 376 MHz 19F-NMR spectrum (# f32537.GID.401) were acquired at room temperature using a Varian UN1TYplus 400 FT-NMR spectrometer. The p-HFX was used as a _H/19F-NMR cross integration standard to permit the cross correlation of the relative lH and l_F signal intensities for evaluation of the overall sample composition. RESULTS: The combined NMR spectral data indicated the sample of 127498-80 consisted of a high purity form of the nominal isomeric product mixture, C_2_-SO3 (') M_+),where 'n' was mainly 6 and where the metal cation ['M (+)'] was assumed to be K(+). Trace-levels of probable aliphatic hydrocarbon impurity components were also observed and quantified. The qualitative and quantitative compositional results that were derived from the s__ trial II-I/19F-NMR cross integration analysis are summarized in TABLE-l on the following page. Any water that may have been present in the sample was ignored for calculation purposes. The relative weight percent concen_ations shown in TABLE-1 should be very close to their respective absolute weight percent values. Trace amounts of other unidentified impurity components were also detected in the NMR spectra, but additional work would be needed in an effort to assign or quantify the unassigned impurities. Copies of the NMR spectra are attached with the paper copy of this report for your reference. If you have any questions about these results, or if any further work is needed, please let me know. Tom Kestner c: Joel Miller George Moore Rick Payfer File Reference:dh32537.GID.do/93 Page I of 2 418-028:PAGE F-3 April 15, 2002 3M SMMD Analytical Lab Request # GID:32537 TABLE-I Sample: 127498-80: PFHS made by George Moore and to be used for toxicological testing. Overall Quantitative Compositional Results by IH/_gF-NMR Cross Inter;ration Analysis Identified Components * _gF/_H-NMR Relative Wt.% Concentrations (single trial measurement) CF3CF2CF2CF2CF2CF2SO3 (') K (+) 88.94% (CF3)2-CF-(CF2)3-SO3 (')K {*) 6.83% CF3CF2-CF(CF_)-CF2CF2-SO3 (') K(+) 2.75% CF3CF2CF2CF2-CF(CF_)-SO3 (') K (+) 0.83% CF3CF2CF2CF(CFa)-CF2-SO_ (') K (+) 0.56% (CF3)rC-CF2-CF2SO_ (') K (+) 0.078% Probable C__H_+2hydrocarbons 0.0034% Traceamountsof otherunassignedcomponemswerealso detectedin theNMR spectra. StudyNo.FACT-TCR008 Page2 of2 APPENDIX G ANALYTICAL AND BIOANALYTICAL REPORT 418-028:PAGE G-1 ANALYTICAL PHASE STUDY TITLE Oral (Gavage) Combined Repeated Dose Toxicity Study ofT-7706 Reproduetion_evelopmental Toxicity Screening Test with the DATA REQUIREMENTS OECD Principles on Good Laboratory Practice. ENV/MC/CHEM(98)l 7, U.S. Food and Drug Administration 21 CFR Part 58, MHW Good Laboratory Practice Standard for Safety Studies on Drugs Ordinance Number 21, March 26, 1997 STUDY DIRECTOR Raymond G. York ANALYTICAL REPORT COMPLETION July 21, 2003 DATE PERFORMING LABORATORY Exygen Research 3058 Research Drive State College, PA 16801 Phone: 814-272-1039 TESTING FACILITY Argus Research 905 Sheehy Drive, Building A Horsham, PA 19044-1297 Phone: 215-443-8710 STUDY SPONSOR 3M Corporate Toxicology Building 220-2E-02 St. Paul, MN 55144-1000 PROJECT Protocol Number: 418-028 Sponsor's Study Number: T-7706.1 Exygen Study Number: 023-072 Total Pages: 153 418-028:PAGE G-2 ' Exygen Study No.: 023-072 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Exgyen Study Number 023-072, entitled "Oral (Gavage) Combined Repeated Dose Toxicity Study of T-7706 with the Reproduction/Developmental Toxicity Screening Test," conducted for 3M Corporate Toxicology, was performed in compliance with U.S. Food and Drug Administration Good Laboratory Practice Regulations 21 CFR Part 58, OECD Principles on Good Laboratory Practice. ENV/MC/CHEM(98)17 and MHW Good Laboratory Practice Standards for Safety Studies on Drugs Ordinance Number 21 by Exygen Research. oPrincipal Investigator Exygen Research _yy___,v_.__ Argus Research John Butenhoff Study Monitor 3M Corporate Toxicology oato Date2"'Tac zoo.a Date Exygen Research Page 2 of 153 418-028:PAGE G-3 Exygen Study No.: 023-072 QUALITY ASSURANCE STATEMENT Exygen Research's Quality Assurance Unit reviewed Exygen Study Number 023-072, entitled, "Oral (Gavage) Combined Repeated Dose Toxicity Study of T-7706 with the Reproduction/Developmental Toxicity Screening Test". All reviewed phases were inspeCted for conduCt according to Exygen Research's Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Exygen Principal Investigator and Management and to the Study Director. Phase 1. Protocol Review Date Inspected 09/24/02 Date Reported to Date Reported to Principal Exygen Date Reported to Investigator Management StudyDirector 11/20/02 11/22/02 01/20/03 2. Extraction, Fortification 09/24/02 3. Raw Data Review and Draft Analytical Report Review 02/03-07/03 and 02/10-11/03 4. Final Analytical Report Review 07/21/03 11/21/02 02/24/03 07/21/03 12/18/02 02/28/03 07/21/03 01/20/03 03/07/03 07/21/03 //// urance Audi_./ Exygen Research Page 3 of 153 418-028:PAGE G-4 ExygenStudy No.: 023-072 CERTIFICATION OF AUTHENTICITY This report, for Exygen Study Number 023-072, is a true and complete representation of the raw data for the study. Submitted by: Exygen Research 3058 Research Drive State College, PA 16801 (814) 272-1039 Principal Investigator, Exygen: /d'ohn Flaherty / " Vice President Exygen Research Exygen Research Facility Management: Exygen Research -// -/9 Study D_ector, Argus Research: _:_/_rln_ol_na__lkk3, York- I/ Associate Director of Rese_'arc_ Argus Research Study Monitor, 3M: John Butenhoff 3M Corporate Toxicology Date Date 2,-,J6t t. -03 Date Date Exygen Research Page4 of 153 418-028:PAGE G-5 Exygen Study No.: 023-072 STUDY IDENTIFICATION Oral (Gavage) Combined Repeated Dose Toxicity Study ofT-7706 with the Reproduction/Developmental Toxicity Screening Test PROTOCOL NUMBER: 418-028 SPONSOR'S STUDY NUMBER: EXYGEN STUDY NUMBER: TYPE OF STUDY: T-7706.1 023-072 Residue SAMPLE MATRIX: Rat Liver, Serum, and Plasma TEST SUBSTANCE: Perfluorohexanesulfonate (PFHS) SPONSOR: 3M Corporate Toxicology Building 220-2E-02 St. Paul, MN 55144-1000 STUDY DIRECTOR: Raymond G. York Argus Research 905 Sheehy Drive, Building A Horsham, PA 19044-1297 TESTING FACILITY: STUDY MONITOR: Argus Research 905 Sheehy Drive, Building A Horsham, PA 19044-1297 John Butenhoff 3M Corporate Toxicology Building 220-2E-02 St. Paul, MN 55144-1000 PERFORMING LABORATORY: Exygen Research 3058 Research Drive State College, PA 16801 ANALYTICAL PHASE TIMETABLE: Study Initiation Date: Analytical Start Date: Analytical Termination Date: Analytical Report Completion Date: 03/26/02 09/16/02 11/13/02 07/21/03 Exygen Research Page 5 of 153 418-028:PAGE G-6 Exygen Study No.: 023-072 PROJECT PERSONNEL The Study Director for this project was Raymond G. York at Argus Research. The following personnel from Exygen Research were associated with various phases of the study: Name John Flaherty Karen Risha Paul Connolly Xiaoming Zhu Lawrence Ord Rickey Kelley Amy Sheehan Emily Decker Mark Neeley Title Vice President Scientist Technical Lead-LC/MS Technician Sample Custodian Sample Custodian Technician Scientist Scientist Exygen Research Page 6 of 153 418-028:PAGE G-7 ' Exygen Study No.: 023-072 TABLE OF CONTENTS TITLE PAGE ....................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT .............................. 2 QUALITY ASSURANCE STATEMENT .......................................................................... 3 CERTIFICATION OF AUTHENTICITY ............................................................................ 4 STUDY IDENTIFICATION ............................................................................................... 5 PROJECT PERSONNEL .................................................................................................... 6 TABLE OF CONTENTS ..................................................................................................... 7 LIST OF TABLES ............................................................................................................... 8 LIST OF FIGURES ............................................................................................................. 9 LIST OF APPENDICES .................................................................................................... 10 1.0 SUMMARY ................................................................................................................ I1 2.0 OBJECTIVE ............................................................................................................... ll 3.0 INTRODUCTION ...................................................................................................... 11 4.0 ANALYTICAL TEST SAMPLES ............................................................................. 12 5.0 REFERENCE MATERIAL ........................................................................................ 13 6.0 DESCRIPTION OF ANALYTICAL METHOD ........................................................ 13 6.1. Extraction Procedure ............................................................................................... 13 6.2 Preparation of Standards and Fortification Solutions ............................................... 14 6.3 Chromatography ....................................................................................................... 14 6.4 Instrument Sensitivity ............................................................................................... 14 6.5 Description of LC/MS/MS Instrument and Operating Conditions ........................... 15 6.6 Quantitation and Example Calculation ..................................................................... 15 7.0 EXPERIMENTAL DESIGN ...................................................................................... 17 8.0 RESULTS ................................................................................................................... 18 9.0 CONCLUSIONS ......................................................................................................... 18 10.0 RETENTION OF DATA AND SAMPLES ............................................................. 19 Exygen Research Page 7 of 153 418-028:PAGE G-8 Exygen Study No.: 023-072 LIST OF TABLES Table I. Summary of PFHS in Control Rat Plasma Samples..........i .......................... ..21 Table II. Summary of PFHS in Control Rat Serum Samples ...................................... .21 Table HI. Summary of PFHS in Control Rat Liver Samples ......................................... .21 Table IV. Summary of PFHS Fortification Recoveries in Rat Plasma. ......................... .22 Table V. Summary of PFHS Fortification Recoveries in Rat Serum .......................... 22 Table VI. Summary of PFHS Fortification Recoveries in Rat Liver ............................. .23 Table VII. Summary of PFHS Residues in Rat Plasma Samples ................................. 24 Table VIII. Summary of PFHS Residues in Rat Serum Samples ...................................... 26 Table IX. Summary of PFHS Residues in Rat Liver Samples. ..................................... .28 Table X. Summary of PFHS Residues in Dosing Solutions ........................................ .40 Table XI. Summary of PFHS Residues in Stability Samples ...................................... .41 Table XII. Summary of PFHS Residues in Homogeneity Samples ................................ .42 Exygen Research Page 8 of 153 418-028:PAGE G-9 Exygen StudyNo.: 023-072 LIST OF FIGURES Figure 1. Typical Calibration Curve for PFHS .............................................................. 44 Figure 2. Chromatogram Representing a 0.1 ng/mL standard for PFHS ....................... 45 Figure 3. Chromatogram Representing a Control Rat Plasma Sample for PFHS (Exygen ID 0202913 Control, Data Set: 091602B) ........................................ 46 Figure 4. Chromatogram Representing a Control Rat Serum Sample for PFHS (Exygen ID: 0202987 Control, Data Set: 091902B) ...................................... 47 Figure 5. Chromatogram Representing a Control Rat Liver Sample for PFHS, (Exygen ID: 0202877 Control, Data Set: 100202A) ..................................... 48 Figure 6. Chromatogram Representing Control Rat Plasma Sample Fortified with 10 ppb ofPFHS (Exygen ID: 0202913 Spk A, Data Set: 091602B) ......49 Figure 7. Chromatogram Representing Control Rat Serum Sample Fortified with 10 ppb ofPFHS (Exygen ID: 0202987 Spk A, Data Set: 091902B) ......50 Figure 8. Chromatogram Representing Control Rat Liver Sample Fortified with 10 ppb ofPFHS (Exygen ID: 0202877 Spk A, Data Set: 100202A) ..... 51 Figure 9. Chromatogram Representing Rat Plasma Sample Analyzed for PFHS (Exygen ID: 0201816, Sponsor ID: 19176 GROUP I, Data Set: 091602B) ..52 Figure 10. Chromatogram Representing Rat Serum Sample Analyzed for PFHS, DF=I 0 (Exygen ID: 0201863, Sponsor ID: 19079 GROUP II, Data Set: 091902BR) ...................................................................................... 53 Figure 11. Chromatogram Representing Rat Liver Sample Analyzed for PFHS (Exygen ID: 0202050, Sponsor ID: 19018 GROUP II Male 1, Dam Set: 100202A) ................................................................................................. 54 Exygen Research Page9 of 153 418-028:PAGE G-10 Exygen StudyNo.: 023-072 LIST OF APPENDICES Appendix A Study Protocol 418-028 (Exygen Study No. 023-072) and Amendments, Deviation andNote to File .................................................. 55 Appendix B Analytical Method: Method of Analysis for the D_nnination of Pcl-fluorohexanesulfonate (PFHS), Perfluorooctanesulfonate (PFOS) and Pentadecafluorooctanoic Acid (PFOA) in Rat Liver, Serum and Urine Revision 1 (Exygen Method No. ExM-023-071 Revision 1) ................. 110 ExygenResearch Page 10 of 153 418-028: PAGE G- 11 Exygen Study No.: 023-072 1.0 SUMMARY Exygen Resarch extracted and analyzed rat liver, serum, and plasma samples for the determination of perfluorohexanesulfonate (PFHS) according to Exygen Method ExM023-071 Revision 1 (Appendix B). The limit of quantitation for PFHS in rat liver was 10 ng/g and 10 ng/mL in rat serum and plasma. The LOQ for each matrix was determined in a method validation study performed at Exygen (Exygen Study No: 023-073). PFHS in the rat plasma samples ranged from non-detected levels to 237,000 ng/mL. PFHS in the rat serum samples ranged from non-detected levels to 189,000 ng/mL. PFHS in the rat liver samples ranged fi'omnon-detected levels to 675,000 ng/g. The average percent recoveries + standard deviations for PFHS in rat plasma, serum, and liver samples were 84% + 7%, 95% + 12%, and 86% + 13%, respectively. PFHS residues in the dosing solutions, stability samples and homogeneity samples were all within 70% to 125% oft.heir known concentrations. 2.0 OBJECTIVE The objective of the analytical part of this study was to determine levels of perfluorohexanesulfonate (PFHS) in specimens of rat liver, serum and plasma samples, bulk test substance, concentration and homogeneity formulations, and stability solutions according to Protocol 418-028 (Appendix A). 3.0 INTRODUCTION This report details the results of the analysis for the determination of PFHS in rat liver, serum and plasma samples, bulk test substance, concentration and homogeneity formulations, and stability solutions using the analytical method entitled, "Method of Analysis for the Determination of Perfluorohexanesulfonate (PFHS), Perfluorooctanesulfonate (PFOS) and Pentadecafluorooctanoic Acid (PFOA) in Rat Liver, Serum and Urine Revision 1." The study was initiated on March 26, 2002, when the study director signed protocol number 418-028. The analytical start date was September 16, 2002, and the analytical termination date was November 13, 2002. Exygen Research Page 11 of 153 418-028:PAGE G-12 ' Exygen Study No.: 023-072 4.0 ANALYTICAL TEST SAMPLES The control rat liver (Exygen ID 0202877) used for the matrix blanks and matrix fortifications was received frozen on dry ice on August 7, 2002 from Pel-Freez Biologicals, Rogers, Arkansas. The control rat plasma (Exygen ID 0202912 and 0202913) used for the matrix blanks and matrix fortifications was received frozen on dry ice on August 8, 2002 from Pel-Freez Biologicals, Rogers, Arkansas. The control rat serum (Exygen ID 0202987) used for the matrix blanks and matrix fortifications was re_'ived frozen on dry ice on August 15, 2002 from Pel-Freez Biologicals, Rogers, Arkansas. Dosage solution/suspension samples (Exygen ID 0201753-0201755) were received refrigerated at Exygen from Argus Research on May 29, 2002. Dosage solution/suspension samples (Exygen ID 0201757-0201758) were received refrigerated at Exygen from Argus Research on June 11, 2002. These samples were logged in by Exygen personnel and placed in refrigerated storage. Bulk TA/S sample (Exygen Research on June 11, 2002. in refrigerated storage. ID 0201756) was received ambient at Exygen from Argus This sample was logged in by Exygen personnel and placed Median liver lobe (Exygen ID 0201759-0201787), pooled fetal liver (Exygen ID 0201788-0201800), plasma (Exygen ID 0201801-0201860), pooled fetal serum (Exygen 1:I30201861-0201874), pooled pup serum (Exygen ID 0201875-0201924), and pup livers (Exygen ID 0201925-0202408) were received frozen on dry ice at Exygen from Argus Research on June 18, 2002. These samples were logged in by Exygen personnel and placed in frozen storage. Median liver lobe (Exygen ID 0203650) and pooled fetal liver (Exygen ID 0203651) were received frozen on dry ice at Exygen from Argus Research on September 11, 2002 and logged in by Exygen personnel and placed in frozen storage. Prepared formulations (Exygen ID 0203923-0203962) were received fi'ozen from 3M on September 19, 2002 and logged in by Exygen personnel and placed in frozen storage. Sample log-in and chain of custody information can be found in the raw data package associated with this study. Storage records will be kept at Exygen Research. Exygen Research Page 12 of 153 418-028:PAGE G- 13 Exygen Study No.: 023-072 5.0 REFERENCE MATERIAL The analytical standard PFHS was received at Exygen on January 26, 2000 from 3M Environmental Technology and Services. The available information for the reference material is listed below. The reference material was stored frozen. Compound PFHS Exygen Inventory No.. SP244 TCR No. SE036 Purity (%) Expiration Date 99.99 01/01/ 10 The molecular structures of test substance is given below: Name: PFHS Chemical Name: Pertluomhexanesulfonate Molecular Weight: 399 FFF /F/F|F _ F_S03 FF F 6.0 DESCRIPTION OF ANALYTICAL METHOD The analytical method ExM-023-071 Revision 1 was used for this study. 6.1. Extraction Procedure A 1 mL aliquot of the serum and plasma and 1 g of liver were used for the extraction procedure for the laboratory controls and fortifications. Due to insufficient sample size, a 100 laL aliquot oft_heserum and plasma and 0.1 g of the liver were used for the extraction procedure for the study samples. After fortification of appropriate samples, the serum samples were brought up to 20 mL with Type I Water and the liver samples were brought up to 10 mL with Type I Water. The serum samples were vortexed for ~ 1 minute and the liver samples were homogenized with a tissuemizer for - 1 minute. An .aliquotof one milliliter was transferred from each sample and 5 mL of aeetonitrile was added and the samples were shaken for ~ 20 minutes. ]'he samples were centrifuged and the supernataut was decanted onto a conditioned SPE column. Then the samples were eluted with 2 mL of methanol. Each sample was analyzed by LC/MS/MS electrospray. An extraction procedure was not necessary for the bulk test substance, concentration and homogeneity formulations, and the stability solutions. 10 mg of the bulk test substance was weighed and brought to volume with methanol in a 100-mL volumetric flask. The ExygenResearch Page 13 of 153 418-028:PAGE G- 14 Exygen StudyNo.: 023-072 sample was then diluted 100000 times with methanol to fit in the range of the analytical curve and analyzed by LC/MS/MS electrospray. The concentration and homogeneity formulations and the stability solutions were all diluted appropriately with methanol and analyzed by LC/MS/MS electrospray. 6.2 Preparation of Standards and Fortification Solutions A stock standard solution of PFHS was prepared on August 20, 2002 as specified in Exygen method ExM-023-071 Revision 1. The stock standard solution was prepared at a concentration of 100 I.tg/mL by dissolving 10 mg of each standard (corrected for purity only) in methanol. From this solution, a 1.0 _tg/mL fortification standard solution was prepared by taking 1 mL of the stock and bringing the volume up to 100 mL with methanol. A 0.1 _tg/mL fortification standard was prepared by taking 10 mL of the 1.0 _tg/mL fortification standard and bringing the volume up to 100 mL with methanol. A set of standards containing PFHS was prepared via dilution of the 0.1 I.tg/mL and various calibration solutions in the following manner: Initial Cone. (l_g/mL) 1 0.1 0.1 0.1 0.005 0.002 0.001 l of PFHS Volume (mL) 5 2 1 10 10 10 Diluted to (mL) 100 100 100 100 100 100 Final Cone. (_tg/mL) 0.005 0.002 0.001 0.0005 0.0002 0.0001 The stock standard solution and all fortification and calibration standard solutions were stored in a refrigerator (4 :t: 2C) when not in use. Documentation of standard preparation can be found in the raw data assoeiated with this report. 6.3 Chromatography Quantification of PFHS was accomplished by LC/MS/MS eleetrospray. The retention time of PFHS was ~ 8.3 rain. Peaks above the LOQ were not detected in any of the control samples corresponding to the analyte retention time. 6A Instrument Sensitivity The smallest standard amount injected during the chromatographic run had a concentration of 0.0001 _tg/mL of PFHS. Exygen Research Page 14 of 153 418-028:PAGE G-15 Exygen Study No.: 023-072 6.5 Description of LC/MS/MS Instrument and Operating Conditions Instrument: Mieromass Quattro Ulfima (Mieromass) Interface: Electrospray (Micromass) Computer: COMPAQ Professional Workstation AP200 Software: Windows NT, Masslynx 3.3 HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser liP Autosampler HP Column Oven HPLC Column:Genesis C 8(Jones Chromatography), 2.1 mmx Column Temp.: 35 C Injection Vol.: 15 _tL Mobile Phase (A): 2 mM Ammonium Acetate in type I water Mobile Phase (B): Methanol 50 ram, 4bt Time %A 0.0 90 2.0 90 5.0 10 9.0 10 9.5 0 14.0 0 14.5 90 20.0 90 %13 Flow Rate (mL/min) 10 0.3 10 0.3 90 0.3 90 0.3 100 0.3 100 0.3 10 0.3 10 0.3 Ions monitored: Ana138_g PFHS Mode Negative Transition Monitored 399 --_ 80 Approximate Retention Time (min) -8.3 rain. 6.6 Quantitation and Example Calculation Fifteen mieroliters of sample or calibration standard were injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x fit weighted linear regression) by Masslynx software using six concentrations of standards. The concentration was determined from the equations below. Equation 1 calculated the amount of aualyte found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Masslynx software Exygen Research Page 15 of 153 418-028:PAGE G-16 Exygen Study No.: 023-072 program. Then Equation 2 calculated the amount of analyte found in ng/g for liver and ng/mL for serum and plasma. Equation 1: Analyte found (ng/mL) = (Peak area - intercept) x DF x AF slope Where: AF = Aliquot Factor DF = Dilution Factor Equation 2: Analyte found (ppb, ng/g for liver and ng/mL for serum and plasma) = analyte found (ng/mL) x FV (mL) sample volume (mL) or sample weight (g) Where: FV --"Final Volume For samples fortified with known amounts of PFHS prior to extraction., Equation 3 calculated the percent recovery. Equation 3: Recovery (%) = ((analvte found (lalab)- analyte in control _pb)) amount added (ppb) x 100% To find the total analyte found corrected for the salt content Equation 4 was used. Equation 4: Total Analyte Found Corrected (ppb) = analyte found (ppb) x salt correction factor Where the salt correction factor = 0.91. An example of a calculation using an actual sample follows: Rat liver sample Exygen ID 0202877 Spk A (Set: 100202A), fortified at 10 ng/g where: peak area intercept slope dilution factor = 2125 = 41.0365 = 4728.12 = 1 ppb added (fort level) = avg. amt in controls = final volume = 10 0 (Not Detected) 2 mL aliquot factor sample weight = 10 = 1.0 g Exygen Research Page 16 of 153 418-028:PAGE G-17 , Exygen StudyNo.: 023-072 From equation 1: Analyte found (ng/mL) From equation 2: Analyte found (ppb) From equation 3: % Recovery From equation 4: Analyte Found Corrected (ppb) = [2125 - 41.0365] 1 10 4728.12 = 4.41 ng/mL = (4.41 ng/mL 2 mL) (1.0g) = 8.82 ppb = ((8.82 ppb - 0 ppb) 100% 10 ppb = 88% = 8.82 ppb x 0.91 = 8.03 ppb 7.0 EXPERIMENTAL DESIGN Each set of samples (liver, serum or plasma) consisted of one matrix blank, two matrix blanks fortified at known concentrations, and - 20 samples. Each sample was extracted using the method and then analyzed in duplicate. ExygenResearch Page 17 of 153 418-028:PAGE G-18 Exygen Study No.: 023-072 8.0 RESULTS The PFHS found in the control rat plasma, serum, and liver samples are listed in Tables I-HI. Peaks were not detected in any of the control plasma or serum samples corresponding to the analyte retention time. Some peaks were detected in the control liver samples corresponding to the analyte retention time; however, the peaks detected were all below the LOQ of 10 ppb. Fortification recoveries for PFHS in the rat plasma, serum, and liver samples are detailed in Tables IV-VI. The average percent recoveries + standard deviations for PFHS in rat plasma, serum, and liver samples were 84% + 7%, 95% + 12%, and 86% _+ 13%, respectively. PFHS in the rat plasma samples ranged from non-detected levels to 237,000 ng/mL. Individual results are listed in Table VII. PFHS in the rat serum samples ranged from non-detected levels to 189,000 ng/mL. Individual results are listed in Table VIII. PFHS in the rat liver samples ranged from non-detected levels to 675,000 ng/g. Individual results axe listed in Table IX. PFHS residues in the dosing solutions ranged from 88% to 125% of their known concentrations. The results are listed in Table X. PFHS residues found in the stability samples ranged from 91% to 122% of their known concentrations. The results are listed in Table XI. PFHS residues found in the homogeneity samples ranged from 70% to 116% of their known concentrations. The results are listed in Table XII. 9.0 CONCLUSIONS The rat liver, serum, and plasma samples were successfully extracted and analyzed for PFHS according to analytical method ExM-023-071 Revision 1. The bulk test substance, concentratiaonnd homogeneityformulationasn,d stabilitsyolutionwsere also successfulalnyalyzefdorPFHS accordintgoanalyticmaelthodExM-023-071Revision I. Exygen Research Page 18 of 153 418-028:PAGE G- 19 Exygen Study No.: 023-072 10.0 RETENTION OF DATA AND SAMPLES When the final analytical report is complete, all original paper data generated by Exygen Research will be shipped to the sponsor. This does not include facility-specific raw data such as instrument or temperature logs. Exact copies of all raw data, as well as a signed copy of the final analytical report and all original facility-specific raw data, will be retained in the Exygen Research archives for the period of time specified in 21 CFR Part 58, OECD ENV/MC/CHEM(98)17, and MHW Ordinance Number 21. Retained samples of reference substances are archived by the sponsor. ExygenResearch Page 19 of 153 418-028:PAGE G-20 Exygen Study No.: 023-072 TABLES Exygen Research Page 20 of 153 418-028:PAGE G-21 Exygen Study No.: 023-072 Table I. Summary of PFHS in Control Rat Plasma Samples Sponsor ID LOT 17824 LOT 17824 LOT 05024 LOT 17824 Exygen ID 0202913 Control 0202913 Control 0202912 Control 0202913 Control Set Number 091602AR 091602B 091802A 091902A PFHS Found _ppb) ND ND ND N'D Table II. Summary of PFHS in Control Rat Serum Samples Sponsor ID 36119-3 36119-3 36119-3 36119-3 36119-3 Exygen ID 0202987 Control 0202987 Control 0202987 Control 0202987 Control 0202987 Control Set Number 091902B 092002A 092002B 092302A 101102/k PFHS Found (ppb) ND ND ND ND ND Table HI. Summary of PFHS in Control Rat Liver Samples Sponsor El) LOT 21824 LOT 21824 LOT 21824 LOT 21824 LOT 21824 LOT 21824 LOT 21824 lOT 21824 LOT 21824 LOT 21824 LOT 21824 LOT 21824 LOT21824 LOT 21824 LOT 21824 LOT 21824 Exygen ID 0202877 Control 0202877 Control A 0202877 Control A 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 Control 0202877 control 0202877 cohtrol 0202877 Control 0202877 Control Set Number 09280ZA 092602A 092602B 092702A 092702B 100202A 100102A 100302A 100402A 100402BR 100902AR 100902BR 101002A 101002B 101102AR 101402A PFHS Found (ppb) ND ND 2.32 (NQ) 3.84 (NQ) 3.85 (NQ) ND ND ND ND 2.09 (NQ) 3.95 0NQ) 2.01 0NQ) 1.85 (NQ) 3.95 (NQ) 4.73 (NQ) 3.37 (NQ) ND = Not Detected (Area less than the lowest concentration of the calibration standards (0.1 ng/mL)) NQ = Not Quantifiable (Area is greater than 0.1 ng/mL but less than LOQ (10 ng/mL)) Exygen Research Page 21 of 153 418-028:PAGE G-22 ' Exygen StudyNo.: 023-072 Table IV. Summary of PFHS Fortification Recoveries in Rat Plasma Sponsor ID LOT 17824 LOT 17824 19176 GROUP 1 LOT 17824 LOT 17824 19176 GROUP I LOT 05024 LOT 05024 19076 GROUP I LOT 17824 LOT 17824 19076 GROUP I Exygen m 0202913 Spk A 0202913 Spk B 0201801 Spk C 0202913 SpkA 0202913 Spk B 0201816 Spk C 0202912 Spk A 0202912 Spk B 0201846 Spk C 0202913 Spk A 0202913 SpkB 0201831 Spk C Set Amount Numbe r 091602A 091602A 09160ZA 091602B 091602B 091602B 091802A 091802A 091802A 091902A 091902A 091902A Added _pb) 10 50 5000 10 50 5000 I0 50 5000 10 50 5000 Relative Standard Standard Average: Deviation: Deviation: % Recovery 91 84 77 86 84 78 88 86 89 71 96 82 84 7 8 Table V. Summary of PFHS Fortification Recoveries in Rat Serum Sponsor ID 36119-3 36119-3 19076 GROUP I 36119-3 36119-3 19012 GROUP I 36119-3 36119-3 19036 GROUP II 36119-3 36119-3 19005 GROUP IV 36119-3 36119-3 Exygen ID 0202987 Spk A 0202987 Spk B 0201861 Spk C 0202987 Spk A 0202987 Spk B 0201875 Spk C 0202987 Spk A 0202987 Spk B 0201890 Spk C 0202987 SpkA 0202987 Spk B 0201905 Spk C 0202987 SpkA 0202987 Spk B Set Amount Number Added (ppb) 091902B 10 091902B 50 091902B 5000 092002A 10 092002A 50 092002A 5000 092002B 10 092002B 50 092002BR 5000 092302A 10 092302A 50 101102A 200000 101102A 10 101102A 50 Average: Standard Deviation: Relative Standard Deviation: % Recovery 92 96 94 106 105 90 97 97 120 90 101 88 78 72 95 12 12 Exygen Research Page 22 of 153 418-028:PAGE G-23 Exygen Study No.: 023-072 Table VI. Summary of PFHS Fortification Recoveries in Rat Liver Spenser ID LOT 21824 LOT 21824 19176 GROUP I LOT 21824 LOT 21824 19076 GROUP I LOT 21824 LOT 21824 19076 GROUPI I)T 21824 LOT 21824 19012 GROUP I, Male I LOT 21824 LOT 21824 19021 GROUP I, Male I LOT 21824 LOT 21824 19018 GROUPII, Male 1 LOT 2 i 824 LOT 21824 19041 GROUP I, Male 1 LOT 21824 LOT 21824 19036 GROUP II, Male I LOT 21824 LOT 21824 19003 GROUPHI, Male 1 LOT 21824 LOT 21824 19008 GROUP III, Male I LOT 21824 LOT 21824 19015 GROUP lJI, Male I LOT21824 LOT 21824 19035 GROUP IV, Male 1 LOT 21824 LOT 21824 19040 GROUP IV, Male 2 LOT21824 LOT 21824 19006 GROUPV, Male 1 LOT21824 LOT21824 19020 GROUPV, Male 2 LOT21824 LOT21824 19025 GROUPV, Male I Exygen ID 0202877 Spk A 0202877 Spk B 0201759 Spk C 0202877 Spk A 0202877 Spk B 0201774 Spk C 0202877 Spk A 0202877 Spk B 0201788 Spk C 0202877 Spk A 0202877 Spk B 0201925 Spk C 0202877 Spk A 0202877 Spk B 0201945 SpkC 0202877 SpkA 0202877 SpkB 0202050 SpkC 0202877 SpkA 0202877 Spk B 0201965 SpkC 0202877 Spk A 0202877 Spk B 0202070 Spk C 0202877 Spk A 0202877 Spk B 0202119 SpkC 0202877 Spk A 0202877 Spk B 0202139 SpkC 0202877 Spk A 0202877 Spk B 0202159 SpkC 0202877 Spk A 0202877 SpkB 0202229 Spk C 0202877 Spk A 0202877 Spk B 0202249 SpkC 0202877 SpkA 0202877 SpkB 0202320 SpkC 0202877 Spk A 0202877 SpkB 0202340 SpkC 0202877 SpkA 0202877 SpkB 0202360 Spk C Exygen Research Set Amount Number 092802A Added(ppb) 10 092802A 50 092802A 5000 092602A 10 092602A 50 092602A 5000 09260213 10 092602B 50 0926071:1 5000 092702A 10 092702A 50 092702A 5000 092702B 10 092702B 50 092702B 5000 100202A 10 100202A 50 100202A 5000 100102A 10 100102A 50 i 00102A 5000 100302A 10 100302A 50 100302A 5000 100402A 10 100402A 50 100402A 5000 100402BR 10 100402BR 50 !00402BR 5000 100902AR 10 100902AR 50 100902AR 5000 100902BR 10 100902BR 50 100902BR 5000 101002A 10 101002A 50 101002AR 5000 101002B 10 101002B 50 101002BR 5000 101102AR 10 I01102AR 50 101102AR 25000 10! 402A 10 101402A 50 101402ARR 5000 Average: Standard Deviation: Relative Standard Deviation: % Recovery 83 87 96 125 I 10 100 104 87 111 75 93 94 83 100 109 88 71 76 74 77 76 98 81 77 81 79 62 70 78 86 72 8I 79 72 70 72 96 93 84 97 72 90 88 75 83 95 77 110 86 13 16 Page 23 of 153 418-028:PAGE G-24 Exygen Study No.: 023-072 Table VII. Summary of PFHS Residues in Rat Plasma Samples ' Sponsor ID 19176 GROUP I 19176 GROUP 1" 19177 GROUP 1 19177 GROUP 1" 191"fS GROUP I 19178 GROUP I* 19179 GROUP !I 19179 GROUP II* 19180 GROUP H 19180GROUP 11" 19181 GROUP 11 19181 GROUP lI* 19182 GROUP lrl 19182 GROUP flit, 191_3 GROUP Ill 19183 GROUP m* 19184 GROUP 111 19184 GROUP HI* 19188 GROUP IV 19185 GROUP IV* 19186 GROUP IV 19186 GROUP IV* 19187 GROUP IV 19187 GROUP IV* 19188 GROUP V 19188 GROUP V* 19189 GROUP V 19189 GROUP V* 19190 G_OUP V 19190 GROUP V* 19176 OROUP I 19176 GROUP It. 19177 GROUP I 19177 GROUP 1" 19178 GROUP I 19178 GROUP I* 19179 GROUP H 19179 GROUP ll* 19180 GROUP II 19180 GROUP Ht. 19181 GROUPII 19181 GROUP lit. 191112 GROUP 111 19182 GROUP fli* 19183 GROUP Ill 19183 GROUP In* 19184 GROUP HI 19184 GROUP HI* 19185 GROUP IV 19188 GROUP IV* 19186 GlUtOUP IV 19186 GROUP IV* 19187 GROUP IV 19187 GROUP IV* |9188 GROUP V 19188 GROUP V* 19189 GROUP V 19189 GROUP V* 19190 GROUP V 19190 GROUP V* E'xygea ' ID 0201801 0201801 Dup laj. 0201802 0201802 Dup lnj. 0201803 0201803 Dup inj. 0201804 0201804 Dup lnj. 0201805 0201805 Dup inj. 0201806 0201806 D'up Inj. 0201807 0201807 Dup Inj. 0201808 020181}8 Dup Inj. 0201809 0201809 Dup Inj. 0201810 0201810Dup laj. 0201811 0201811 Dup Inj. 0201812 0201812 Dulp /,nj. 0201813 0201813 D_p Inj. 0.201814. 0201814 Dup Inj. 0201815 0201813 Dgp Inj 0201816 0201816 Dup lnj 0201817 0201817 Dup I_ ff201818 0201818 Dup laj 0201819 0201819 Dup Inj 0201820 0201820 D up lnj 0201821 0201821Dap lnj 0201822 020182.2 D up Inj 0201823 0201823 Dup _j 0201824 0201824 Dup Inj 0_018_ 0201825 Dup lnj 02018.26 0201826 Dup laj 0201827 0201'827 D up lnj 07..01g2g 0201828 D,,_ Inj 0201829 0201829 Dup Inj 0201830 0201830 Pup lai Matrix ,, , PD 14 PI..ASMA I'D 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA I'D 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PI..ASMA PD 14I_,..ASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 P_ PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD I_t PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 P_ PD 14 PI.,ASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLA,.SMA PD 42 PLASMA PD 42 PLASMA PD 42 PLA,.qMA PD 42 PLASMA PD 42 PI.ASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA I'D 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PI.ASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA _ 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA PD 42 PLASMA Collection D_e 4/14/02 4/14/02 4/14/02 4/1_2 4/In/U2 4/14/(12 4/14.'02 4/14/02 4/14102 4/14/02 4/14102 4/14/02 4/14102 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 4/1471)2 41402 4/14/02 4/14/I)2 4/14/I}2 4/14./02 4/14/0"2 4/14402 4/1_02 4/14/U2 4/14/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12,g)2 5/12/02 _12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5,"12.g12 $/12/02 5/12/02 5/12/02 5/12/ff2 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5112/02 5/12/02 $112/02 5/12/02 5/12/02 _q N_ 091602A 091602A 091602A 0916U2A 091602A 091602A 09160ZAR 091602AR 091602AR 091602AR 091602AR 091602AR 091602AR 091602AR 091602AR 09"602AR 091602AR 091602AR 091602AR 091602AR 091602AR 09t602AR 091602AR 091602AR 091602AR 091602AR 091602AR 091602AR 091602A.R 091602AR 09160215 09160211 091602B 091602B 091602B 0916028 091602BR 091602BR 091602BR 091602BR 091602BR 091602BR 091602BR 091602BR 091602BR 091602BR 091602BR 091602BR 091609nR 091602BR 091602BR 091602BR 0916ff2BR 091602BR u'gI607.BR 091602BR 09160.2BR 091602BR 091602BR 091602BR PFHS Fnmul(n_/mL) 182 172 103 108 46.5 _ 43.3 _ 183013 1940_ 15700 15309 20000 20400 58000 58000 68300 69500 117000 115IX}0 70400 71900 126000 125000 237000 228000 1970130 182000 187000 183000 177000 1700(}0 245 256 3112 376 74.7 _ 68.4 _ 57.400 60200 35700 36400 37100 38500 91000 85900 90300 87500 90700 89300 119000 119000 137000 142000 128000 127000 I1_000 188000 224000 225000 191000 193000 MAlthoughthe method used allows for alternatesamplevolumes orweights andall the peakarea responsesare withinthe calibrationcurve limits,theprecisionfor the 0.1 mL or 0.1 g samplesbelow 100 ppb hasnot been validated. *DuplicateInjection Exygen Research Page 24 of 153 418-028:PAGE G-25 Exygen Study No.: 023-072 Table VII (cont'd). Summary of PFHS Residues in Rat Plasma Samples Sponsor ID 19076 GROUP I 19076 GROUP I" 19077 GROUP I 1907"7GP_OUPI* 19078 GROUP I 19078 GROUP 1" 19079 GROUP I1 19079 GROUP H* 19080 GROUP II 190_0 GROUP 11" 19081 GROUP II 19081 GROUP l]* 19082 GROUP HI 19082 GROUP IIl* 19083 GROUp Ili 19083 GROUP HI" 19084 GROUP In 19084 GROUP HI* 19085 GROUP IV 19085 GROUP IV* 19086 GROUP IV 19086 GROUP IV" 19087 GROUP IV 19087 GROUP IV* 19088 GROUP V 19088 GROUP V* 19089 GROUP V 19089GROUP V* 19090 GROUP V 19090 GROUP V* 19076 GROUP I 19076 GROUP I* 19077 GROUP I 19077 GROUP !* 19078 GROUP I 19078 GROUP 1' 19079 GROUP II 19079 GROUP II* 19080 GROUP I1 19080 GROUP I1' 19081 GROUP H 19081 GROUP II* 19082 GROUP Ill 19082 GROUP HI* 19083 GROUP IU 19083 GROUP II1" 19084 GROUP HI 19084 GROUP III" 19085 GROUP IV 19085 GROUP IV" 19086 GROUP IV 19086 GROUP IV* 19087 GROUP IV 190_7 GROUP IV* 19088 GROUP V 19088 GROUP V* 19089 GROUP V 19089 GROUP V" 19090 GROUP V 19090GROUP V* * DuplicateInjection Exygen ID 0201846 0201846 Dtzp lnj 0201847 0201847 Dup Inj 0201848 0201848 Dup laj 0201849 0201849 Dup Inj 0201850 0201850 Dup lnj 0201851 0201851 Dup inj 02018.q2 0201852 Dup Inj 020185"3 0201853 Dup lqj 0201854 0201854 _ l.j 0201855 0201855 Dup lnj 0201856 0201856 Dup Inj 0201857 0201857 Dup laj 0201858 0201858 Dup t.j 0201859 0201859 D_ laj 0201860 0201860 Dup Inj 0201831 0201831 Dup i_j 0201832 0201832 I_p Inj 0201833 0201833 Dup laj 0201834 0201834 Dup Inj 0201835 0201835 Dup Inj 0201836 0201836 D_laj 0201837 0201837 Dup I_j 0201838 0201838 Dup Inj 0201839 0201839 Dup I_ 0201840 0201840 Dup lnj 0201841 0201841 Dap I_j 0201842 0201842 Dup laj 0201843 0201843 Dup laj 0201844 0201844 Dup laj 0201845 0201845 Due In_ Matrix GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLA,qMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 IR.ASMA GD 2 ! PI.,A_MA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 21 PLASMA GD 2t PLASMA GD 21 PLASMA GD 21 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA I'D 14 PLASMA PD 14 PLASMA PD 14 PLASMA i'D 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA PD 14 PLASMA FD 14PLASMA CollecttR Date 5/9/02 5/9/02 5/19/02 5/19/02 5/7102 5/7/02 5/9/02 5/9/02 5/9/02 5/9/02 5/10/02 5/10/02 5/9/02 5/9/02 5/7/02 5/7/02 5/9/02 5/9/02 $/6/02 5/6q)2 51&q)2 51(d02 51fd02 5/fJ02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 4/14/02 4/14/02 4/14/02 4/14/02 4114/02 4/14/02 4114/02 4114/02 4/14/02 4/14/02 4/14/02 4114/02 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 4114/02 4/14/02 4/141402 4/14402 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 4/14/02 Set Number 091802A 091802A 091802A 091802A 091802A 091802A 091802AK 091802AR 09 I802AR 091802AR 091802AR 091802AR 091802AR 091802AR 091802AR 091802AR 091802AR 09180ZAR 091802AR 09180ZAR 091802AR 091802AR 09 ! 802AR 091802AR 091802AR 091802AR 091802AR 091802AR 091802AR 091802AR 091902A 091902A 091902A 091902A 091902A 091902A 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AK 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR 091902AR PFHS Femd (a_, ,,) ND ND ND ND ND ND 4150 4000 3240 3230 2640 2670 7150 7280 18000 18100 6490 6900 24800 25400 40700 40800 32500 32300 72600 73600 53200 54400 52200 52800 ND ND 386 398 ND ND 3800 3530 2600 2630 2040 2090 10400 10100 13100 14000 $$40 5980 16900 17900 24800 25200 19200 20000 43900 43000 47100 48200 34000 35900 ND -- Not Detected(Area less thanlowest calibrationstandardof 0.1 ng/mL) Exygen Research Page 25 of 153 418-028:PAGE G-26 Exygen Study No.: 023-072 Table VIII. Summary of PFHS Residues in Rat Serum Samples Sponsor ID 19076 GROUPI 19076 GROUP P 1907_ GROI._ I 19078 GROUP I* 19079 GROUP II 19079 GROUP n* 19080 GROUP II 19080 GROUP II* 19081 GROUP II 19_1 GROUP I1" 19082 GROUP I11 19082 GROUP m* 190_3 GROUP m 19083 GROUP rl/* 19084 GROUP wI 190114GROUP I11. 19085 GROUPIV 19085 GROUP IV* 19086 GROUPIV 19086 GROUP IV* 19087 GROUP IV 19087 GROUP[V* 19088 GROUP V 19088 GROUP V* 19089 GROUP V 19089 GROUP V* 19090 GROUP V 19090 GROUP V* 19012 GROUP 1 19012 GROUPI* 19019 GROUPI 19019 GROUP I* 19021 GROUP I 19021 GROUP I* 19023 GROUP i 19023 GROUP i* 19041 GROUP I 19041 GROUPI* 19042 GROUP I 19042 GROUP I* 19044 GROUP 1 19044GROUP 1" 19050 GROUP I 19050 GROUPI* 19053 GROUT I 19053 GROUP I* 19065 GROUP 1 19065 GROUP I* 19004 GROUP II 19004 GROUP II* 19009 GROUP ii 19009 GROUP 11" 19016 GROUP II 19016 GROUP II* 19018 GROUP H 19018 GROUP II* 19026 GROUP 11 19026 GROUP ll* 19036 GROUP rl 19036 GROUP U* Exygem Matrix Coli_tiem ID 020|861 0201861 D_ laj 0201862 0201862 Dup kj 0201863 0201863 D_ kaj 0201864 0201864 Dup tnj 0201865 Pooled FeudSe_um Pooled PetalSerum Pooied FetalSerum Pooled FetalSerum Pooled FetalSatma Fooled FeCaSl e_a Pooled Felal Serum poe_d Fel21Semum Pooled Fetal Saum Date 5/9/02 5/9/02 .5/7/02 5/7/02 5/9/02 5/9/02 5/9/02 5/9/02 5/10192 0201865 DUP Inj 0201866 Pooled FetalScrtma Pooled FetalSertma 5/10/02 .5/9/02 0201866 Dup l_j 0201867 0201867 DUP tnj 0201868 0201868 Dup lnj 0201869 0201869 D_ Mj 0201870 0201870 Dup InJ 0201871 0201871 Dwinj 0201872 0201872 Dup _j 0201873 0201873 D_ haj 0201874 0201874 D_ l,,j 0201875 0201875 D_ Inj 0201876 0201876DUPInj 0201877 0201877 Dup I_j 0201878 0201878 Dup I_j 0201879 0201879 Dup lnj 0201880 0201gg0 D_ lrd 0201881 0201881 Dup hlj 0201852 0201882 D_ Inj 0201853 0201883 D_ Inj 0201854 0201884 D_p laj 0201855 0201885 DUPi_ 0201886 0201986 DUPInj 02018117 0201887 Dup lnj 02018118 02DISM Dup J_j 0"201889 0201_89 Dup htj 0201890 Pooled FetalS_ma Pooled FetalScr_m Pooted Fetal _wtma Pooled FetalSerum Pooled FetalSerum Pooled FetalSaem Pooled FetalSerum Poeled FetalS_mn Pooled Fetal Sauna Pooled Petal Serum Pooled Fetal S_mn Pooled FetalSerum Pooled FetalS_em Pooled FetalSeRum Pooled PetalSerma Pooled FetalSerum Pooled FetalSerum Pooled PUP S_am _mh_d Pup S_um Pool_l Pup Samm PoohePdUP_, Pooled P_p Serum PooledPup Saum Pooled Pup S4mm_ Pooled PUP Serma Pookd P up Serum PeoledPup S_um PooledPup Smlm Pookd Pup Sen_ Pooled Pup Serma Pooled Pup Serum Pooled Pup Saran Pcokd P up_ _ P_ _ Pooled Pup Sca-tma Pooled Pup Se_um PooledPu_ Serum Pooled Pup Scax_ Poeied Pup Se_rum PooleclPw_ Sc_mu Pooled Pup Serum Pookd Pup Se_m Peokd Pup Set_m Pookd Pup S_um pooled Pup Serum Pooled Pup Seft_ Pookd Pup S_um Poobzt Pup _ 5/9/02 5/7/02 5/7/02 5_/02 5/9/02 5/6/02 _6/02 $_2 _/02 5/6/02 5/6/02 5/9/02 S/9/02 5/9/02 5/9/02 5/9/02 5_9/02 5/29/02 .5/29/02 5/30/_ 5/30/02 5/29/02 ,_29/02 5/31/02 5/31/02 5/31/02 5/31/02 5/30/02 5/30/02 5/28/02 _2 5/31/02 5/31/02 _30_2 5/30/02 5/30/02 .5/30/02 5/31/02 5/31/02 5/28/02 _ 5/'),8/02 5/211/02 5/29/02 $/29/02 5/'31/0"2 5/31/9"2 f_.8/02 0201890D_ laut PooledPup ,Y_rum 5/28/02 Set Number 091902B 091902.B 0919028 0919028 091902.BK 0919ff2.BR 0919f12BR 0919028R 0919028R 0919028R 0919028R 0919028R 0919028R 091902BR 0919028R 0919028R 0919028R 091902BR 091907.BR 091902BR 091902BR 091902BR 0919028R 0919028R 091902BR 091902.BR 0919(728R 0919028R 092002A 092002A 09200ZA 092007.A 092(X)7_ 092002A 092002A 092002A 09200ZA 092002A 092002A 092002A 092002A 092002A 092002A 092002A 092_2A 09200ZA 092002A 092002A 092002AR. 092002AR 092002AR 092002AR 092002A R 092_2A R 092002A R 0920(Y2AR 092002A R _#21XY2AR 092002_R 092002BR PFHS Feud (hilL) ND ND ND ND 6460 618)0 3940 4040 5280 5370 12600 13500 15700 15700 11700 I 1600 38600 40200 33000 34700 369_0 39200 35700 38400 44900 47900 47900 $1200 ND NEt 65.3 _ 68,9M 81.1 _ 86,0 M 45.6"" 42,7 _ ND ND ND ND ND ND hiD ND ND ND ND ND 9650 9560 7940 7940 8490 8200 g630 8720 5490 5420 6300 6370 MAlthough the method used allows for alternate sample volumes or weights and all the peak area responses are within the calibration curve limits, the precision for the 0.1 mL or 0.1 g samples below 100 ppb has not been validated. * Duplicate Injection ND = Not Detected (Area less than Lowestcalibration standard of 0.1 ng/mL) Exygen Research Page 26 of 153 418-028:PAGE G-27 Exygen Study No.: 023-072 Table VIH (eont'd). Summary of PFHS Residues in Rat Serum Samples Slmamr II) 19037 GROUP I1 19037 GROUP If" 19043 GROUP U 190a[3 GROUP H* 19047 GROUP n 19047 GROUP If* 19048 GROUP H 19048 GROUP IT" 19003 GROUP Ill 19003 GROUP 11I* 19007 GROUP [] 19007 GROUP [] = |9_ GROUP IIl 19006 GROUP In* 19013 GROUP HI 19013 GROUP UI* 19015 GROUP HI 19015 GROUP H* 19017 GROUP m 19017GROUP []" |90_1GgOUP [] 19024 GROUP []t 19029 GROUP [] 19029 GROUP IlI* 19034 GROUP HI 19034 GPA_UP m* 19056 OR.Ot_ HI 19056 GROUP IH* 19005 GROUP IV 19005 GROUP IV* 19035 GROUP IV 19035 GR(YOP IV" 19039GROUP IV 19039 GROUP IV* 19040 GROUP IV 19040 GROUP IV* 19045 GROUP IV 19045 OIUDUP IV'* 19054 GROUP IV 190_ GROUP IV" 190511GROUP IV 19058 GROUP IV* 19062 GROUP IV 19062 GROUP IV* 19063 GROUP IV 19063 GROUP IV* 19066 GROUP IV 19066 GROUP IV* 19001 GROGP V 19001 GROUP V* 19006 GROUP V 19006 GROUP V" 19011 GROUP V 19011 O_ V" 19020 OROUP V 19020 GROUP V* 19022 GROUP V 19022 GROUP V" 19025 GROUP V 19025 OROU_ V" 19027 GROUP V 19_7 GROUP V* 1902g GROUP V 19028 GROUP V= 19030 GROUI' V 19030 GROI_ V* 19031 GROUP V 19031 GROUP V* Exygeu " El) C_01B91 0201091 D_p l_ 0201892 0201892 DUP lnj 020n93 0201893 Dup laj 0201894 0201094 Dup lnj 0_)1895 0201095 Dup lnj 0201896 0201596 DUP lnj 02011197 0201597 Dup lnj 0_018911 020159_ Dup 1_ 0201899 0201899 Dup lm_ (]{201900 0201900DUP Inj 0201901 0201901 Dup kaj 0201902 0201902 DUP Inj 0201903 0201903 D'ap laj 0201904 0201904 Dup Inj 0_01905 0201905 Dep In] 0201906 0201906 Dup I_j 0201907 0201907 Dup Ioj 0201908 020190_ Dup kaj 0201909 0201909 D_ In_ 0201910 0201910 Dup laj 0201911 0201911Dup In_ 0201912 0201912 Dup lnj 0201913 0201913 Dup I_ 0201914 0201914 Dup laj 0201915 0201915 Dup laj 0201916 0201916 Dup In) 0201917 0201917 Dup ht_ 0201918 0201918 Dup Inj 0201919 0201919 Dup I_ 0201920 020192O Dip l.uj 0201921 0_01921D_ Iaj 0_01922 0201922 Dup Inj 0201923 0201923 DUP I_ 0201924 0201924 Dup lmi Matrix Pooled Pup Sezgm Pooledl_pSerum Pooled Pooled Pooled Pooled Pooled Pup Sa_u Pup Scram Pup Se_tm Pup Set_m Pup Serum Pooled Pup Serum Pooled Pup Scr, nm Pooled Pup Seru_ Pooled Pup Sm'mn pooled Pup Serm_ Pooled Pup S a-_mt Pooled PUP Sea, tin Pooled Pup Senzm Poo|ed _ Sen_m Pooled PUP Serum Pooled Pup Sermu Pooled Pup Serum Po_ed P-apScram Pooled. Pup Smm, IN_oled Pup Sm_a Pooled Pap Sertmt PooledPup Sentm Pooled Pup Serum Pooled Pup Sentm P_ted _ Set'l_ pooted Pup Serum Pooled Pup Serum IN_ ed Pup Sm'x_m Pooled Pup Serum Pooled P_ Serum Pml.edP_ Se_m Pooled P_p Senem Pooled Pup Sentm Pooled Pup Sermm Pooled Pup Serum Pooled P'a_ Seru_ Pooled Pup Serum Pooled POp Sere_ pooled Pup S ex'mm Pooled Pup Steam Pooled PW Sa't,tm Pooled Pup Sermm pooled Pup Sentm Pooled Pup Sermm Pooled Pup Seru_ Pooled Pup Scram Poled Pup Set'urn pooled Pup Sm_am Pooled Pup Serum Pooled Pup Sentm P0oled Pup Serum Pooled Pup Serma Pooled Pup Scram Pooled Pup Serum Pooled Pooled Pooled Pooled Pooled Pup Senn Pup Scram Pup Serum Pup Serum Pup Serum Pooled pooled Pooled Pap Serum Pup Serum Pup Serum Poo_edPuO Sen_m Pooled Pap Serum Pooled Pup Sensm Pooled Pup Sm'v_ Collection Date 5/'29102 5/29/02 5/28/02 5r2g/02 5/28/02 5/2N02 5/31/02 5/31/02 5/30/02 5/30/02 5/30/02 5/30/02 .fu31/02 5/31/02 5/29/02 .ru'29_2 5/31/02 5/31/02 5/28/02 5/28/I/2 5/31/02 5/31/02 5/29/02 5/29/02 S/30/02 5/30_2 ._ t_2 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/29002 5/29/02 5/30/02 5/30/02 5/2_/02 5_J_82 5/317{}2 5/31/02 _ 50.8/02 5/31/92 5/31102 5/29/02 5/29/02 5/31/02 $/31/02 5/I _/02 $/13/02 5113/02 5)13/02 5/13/02 5/13/02 5/14/_ 5/14/02 .5/31/02 5/31/02 5/30/02 $/30/02 5/31/02 5/31/02 5r2g/02 5/2g/02 5/31_2 5/31/02 5/2g_2 5/'2W02 * DuplicateInjection Set Namb_r , 092002BR 092002BR 092002 BR 092002BR 092002BR 092002 BR 092002BR 092002BR 092002 BR 092002BR 092002BR 092002 BR 0920ff2BR 092002BR 092002BR 0921_2BI_. 0921Xt2BR 092002BR 092002]RR 092002BR 092002BR 092002BR 092002 BR 0920_BR 092002BR 092002BR 092002 BK 092002BR 09230ZAR 0_2302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 09230ZAR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302A R 092302AR 092302AR 092_02AR 09230_AR 0923_AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR 092302AR PFHS . Fommd _ng/mL) 12800 1_00 5750 5970 8990 8B10 11700 11700 37300 37400 47300 46300 44000 45400 24300 24600 37000 20800 21200 42900 45000 24900 22600 45000 43700 19700 29500 30900 30800 34600 33500 22600 22800 40700 38400 27800 29300 42900 43800 2_8D0 25800 41200 41100 19800 19100 36000 39300 29030 32700 47900 52400 162000 164000 721100 76200 84100 89400 50200 53900 57200 53700 162000 163000 I _1000 159000 71200 76200 Exygen Research Page 27 of 153 418-028:PAGE G-28 Exygen Study No.: 023-072 Table IX. Summary of PFHS Residues in Rat Liver Samples El) 19_76 GROUP I 19176 GROUP I* 19177 GROUP I 19177GROUP I* 19178 GROUP I 19178GROUP I* 19179 GROUP II 19179 GROUP If* 19180 GROUP II 19180 GROUP II" 19181 GROUPII 19181 GROUP 11" 19182 GROUP 11I 19182 GROUP Ill* 19183 GROUP Ill 19183 GROUP []* 19184 GROUP [] 19184 GROUP IlI* 19185GROUP IV 19185 GROUP IV* 19186GROUP IV 19186GROUP IV* 19187 GROUP IV 19187 GROUP IV* 19188 GROUP V 19188 GROUP V* 19189 GROUP V 19189 GROUP V* 19190 GROUP V 19190 GROUP V* 19076 GROUP 1 19076 GROUP 1" 19077 GROUPI 19077 GROUP I* 19078 GROUP1 19078 GROUP 1" 19081 GROUPIIFemale I 1908i GROUP IIFezaale 1" 19079 GROUP Il 19079 GROUP If* 19080 GROUPI1 19080GROUP II* 19082 GROUP Ill 19082 GROUP 1II* 19083 GROUP HI 19083 GROUP lIl* 19084 GROUP Ill 19084 GROUP HI* 19085 GROUP IV 19085 GROUP IV* 19086 GROUP IV 19086 GROUP IV* ID 0201759 .... Median IAv_- Lobe 0201759 Dup Inj 0201760 Median Liver Lobe Median LiverLobe 0201760 Dup Inj 0201761 Median _ Lobe Median LiverLobe 0201761 Dup Inj 0201762 Median Liv_ Lobe Median Liver Lobe 0201762 l_p laj 0201763 Median l..iveaL" obe Median Liver Lobe 0201763 Dup haj 0201764 Median Liver Lobe Median Liver Lobe 0201764 Dup Inj 0201765 MedianLiver Lobe MedianLiver Lobe 0201765 Dup Inj 0201766 MedianLiver Lobe MedianLiver Lobe 0201766 Dup Inj 0201767 Median LiverLobe Median LiverLobe 0201767 Dup I_ 0201768 Median Liver Lobe Medianl..iver Lobe 0201768 Dup lnj 0201769 Median Liv1_Lobe MedianLiver Lobe 0201769 Dup Inj 0201770 Median Liver Lobe Median Liver Lobe 0201770 Dup lnj 0201771 Median Liver lobe Median LiverLobe 0201771 Dup lnj 0201772 Median LiverLobe Median LiverLobe 0201772 Dup Ir_ 0201773 Median Liver Lobe Median Liver Lobe 0201773 Dup Inj 0201774 Median Liver Lobe Median Liver Lobe 0201774 Dup Inj 0201 775 Median LiverLobe Median Liver Lobe 0201775 Dup haj 0201776 Median Liver Lobe Median Livcr Lobe 0201776 Dup Inj 0203650 Median Liver Lobe MedianLiver Lobe 0203650 Dup lnj 0201777 Median Liver Lobe MedianLiver Lobe 0201777 Dup Inj 0201778 MermanLiver Lobe Median Liver Lobe 0201778 Dup Inj 0201779 Median Livez Lobe MedianLiver Lobe 0201779 Dup lnj 0201780 Median Liver Lobe Median Liver Lobe 0201780 Dup Inj 0201781 Median Liver Lobe Median Liver Lobe 0201781 Dup lnj 0201782 Median Lira"Lobe Median Liver Lobe 0201782 Dup In_ MedimtLiver Lobe 0201783 Median Liver Lobe 0201783 Dup Inj Median Live_Lobe , Date 5/|2/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 .5/12/02 5/12/02 5/12/02 5/12/02 5/12/02 5112/02 5/12/02 5/12/02 5/12/02 5/9/02 5/9/02 5/19/02 5/19/02 5/7/02 5/7/02 5110/02 5/10/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/7/02 5/7/02 5/9/02 5/9/02 5/6/02 516/02 5/6/02 5/6/02 Numb.el 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092802A 092602A 092602A 092602A 092602A 092602A 092602A 092602A 092602A 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR Found (ng/mL) 606 576 138 141 326 335 40100 42300 36800 37900 52900 52800 149000 154000 124000 121000 ! 72000 177000 194000 187000 408000 4070(_ 416000 420000 501000 511000 659000 675000 612000 603000 ND ND 82.2'" 82,4 '_ ND ND 628 633 1040 974 762 725 1960 2080 3070 3090 2700 2750 6360 6640 9740 9370 "'Although themethodusedallows for alternatesamplevolumesorweights andall the peakarea responsesarewithinthecalibrationcurvelimits, theprecisionforthe 0.1 mLor 0.1 g samplesbelow 100 ppbhasnot beenvalidated. * DuplicateInjection ND = Not Detected(Area less thanlowestcalibrationstandardof 0.1 ng/mL) Exygen Research Page 28 of 153 418-028:PAGE G--29 Exygen Study No.: 023-072 Table IX. (eont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19087 GROUP IV 19087 GROUP IV* ! 9088 GROUP V 19088 GROUP V* 19089 GROUP V 19089 GROUP V* 19090 GROUP V 19090 GROUP V* 19076 GROUP 1 19076 GROUP I* 19078 GROUP I 19078 GROUP 1" 19081 GROUP II Female 2 19081 GROUP II Female 2* 19079 GROUP H 19079 GROUP 1I* 190g0 GROUP II 19080 GROUP If* 19082 GROUP 111 19082 GROUP ITI* 19083 GROUP HI 19083 GROUP Ill* 19084 GROUP HI 19084 GROUP HI* 19085 GROUP IV 19085 GROUP IV* 19086 GROUP IV 19086 GROUP IV* 19087 GROUP IV 19087 GROUP IV* 19088 GROUP V 19088 GROUP V* 19089 GROUP V 19089 GROUP V* 19090 GROUP V 19090 GROUP V* i9012 GROUP 1, Male 1 19012 GROUP I, Male 1" 19012 GROUP I, Male 2 19012 GROUP I, Male 2* 19012 GROUP I, Male 3 19012 GROUP I, Male 3" 19012 GROUP I, Male 4 19012 GROUP I, Male 4" 19012 GROUP I, Male 5 19012 GROUP l, Male 5* 19012 GROUP L Female 7 19012 GI_OUP I, Female 7* 19012 GROUP I, Female 8 19012 GROUP I, Flanal 8* 19012 GROUP 1, Female 14 ., 19012 GROUP L Female 14" Exygen El) 0201784 0201784 Dup Inj 0201785 0201785 Dup Inj 0201786 0201786 Dup Inj 0201787 0201787 Dup Inj 0201788 0201788 Dup Inj 0201789 0201789 Dup lnj 0203651 0203651 Dup Inj 0201790 0201790 Dup lnj 020179 ! 0201791 Dup lnj 0201792 0201792 Dup Inj 0201793 0201793 Dup Inj 0201794 0201794 Dup Inj 0201795 0201795 Dup Inj 0201796 0201796 Dup Inj 0201797 0201797 Dup Inj 0201798 0201798 Dup Inj 0201799 0201799 Dup Inj 0201800 0201800 Dup Inj 0201925 0201925 Dup lnj 0201926 0201926 Dup Inj 0201927 0201927 Dup Inj 0201928 0201928 Dup lnj 0201929 0201929 Dup lnj 0201930 0201930 Dup Inj 020193 i 0201931 Dup Inj 0201932 0201932 Dup lnj Matrix Median Liver Lobe Median Liver l__be Median Liver Lobe Median Liv_ Lobe Median Liwr Lobe Median Liver Lobe Median Liver Lobe Median Liv_ Lobe Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Pooled Pooled Fetal Liver Fetal Liver Fetal Liver Pooled Fetal Liver Pooled Fetal Live_ Pooled Fetal Live_ Pooled Fetal Liver Pooled Fc'tal Livez Pooled Felal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liv_ Pooled Fetal Liver Pooled Fetal Laver Pooled Fetal Liver Pooled Fetal Laver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pooled Fetal Liver Pup Livea_ Pup Livers Pup Livers Pup Livea_ Pup Livea's Pup Livens Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers PUp Livers PUp Live_ Pup Livers , Pup.Livers Collection Date 516/02 5/6/02 5/9/02 5/9102 5/9/02 5/9/02 5/9102 5/9/02 5/9/02 5/9/02 5/7/02 5/7/02 5/I0/02 5/10/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5n/02 5/7/02 5/9/02 5/9/02 5/6/02 5/6/02 5/6/02 5/6/02 5/6/02 5/6/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/9/02 5/29/02 5/29102 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 Set Number 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602AR 092602B 092602B 092602B 092602B 092602B 092602B 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BK 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092602BR 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 09"2702A 092702A 092702A 092702A 092702A PFHS Found (ngefiazL) 7500 7210 ! 9000 20000 15100 15300 14700 15100 27.7 "_ 25.7 "_ ND ND 789 821 1460 1480 1810 1880 2180 2290 4560 4550 3020 3150 7320 7170 8510 8590 5630 5920 22200 22800 20100 19800 14700 13600 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND AAAlthoughthe methodused allowsfor alternatesamplevolumes or weights and all the peak area responses are within the eah'brationcurvelimits, the precision for the 0.I mL or 0.1 g samplesbelow 100 ppb has not been validated. * Duplicate Injection ND = Not Detected (Area less than lowestcalibrationstandard of 0.1 ng/mL) Exygen Research Page 29 of 153 418-028:PAGE G-30 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Spommr El) 19012 GROUP I, Female 15 19012 GROUP I, Female15" 19012 GROUP I, Female 16 19012 GROUP I,Female 16" 19019 GROUP I, Malc I 19019 GROUP I,Male 1* 19019GROUP I, Male 2 19019 GROUP I,Male 2* 19019 GROUP 1, Male 3 19019 GROUP I, Maic 3* 19019 GROUPI, Male 7 19019 GROUP I, Male 7* 19019 GROUPI, Male 8 19019 GROUP I, Male 8" 19019 GROUP 1, Femalci I 19019 GROUP I, Female 11" 19019 GROUP I, Female12 19019 GROUP I, Female 12' 19019 GROUPI, Female 13 19019 GROUP I, Female 13" 19019 GROUP I, Fenmle 14 19019 GROUP I, Female 14" 19019 GROUP I, Female 15 19019 GROUPI, Female 15' i 902i GROUP I, Male I 19021 GROUPI, Male I* 19021 GROUP I, Male 2 19021 GROUPI, Male 2" 19021 GROUP I, Male 6 19021 GROUPI, Male 6" 19021 GROUP I, Male 8 19021 GROUPI, Male 8" 19021 GROUP I, Malc 9 1902! GROUP I, Male 9* 19021 GROUP I, Female 11 19021 GROUP I, Female11" 19021 GROUP I, Female 12 19021 GROUP I, Female 12" 19021 GROUP I, Female 13 19021 GROUP1, Female 13" 19021 GROUP I, Fcatalc 14 19021 GROUP L Female 14" 19023 GROUP I, Male 1 19023 GROUP I, Male 1" 19023 GROUPI, Male 2 19023 GROUP I,Male 2* 19023 GROUP I, Male 3 19023 GROUPI, Male 3* 19023 GROUP I, Male 6 19023 GROUPI, Male 6" 19023 GROUP I, Male 7 19023 GROUPI, Male 7" Exygen 1D 0201933 0201933 Dup lnj 0201934 0201934 Dup lnj 0201935 0201935 Dup Inj 0201936 0201936 Dap lnj 0201937 0201937 Dup Inj 0201938 0201938 Dup Inj 0201939 0201939 Dup Inj 0201940 0201940 Dup Inj 0201941 0201941 Dup Inj 0201942 0201942 Dup Inj 0201943 0201943 Dup lnj 0201944 0201944 I_p Inj 0201945 0201945 DupInj 0201946 0201946 Dup Inj 0201947 0201947 DUPInj 0201948 0201948 Dup lnj 0201949 0201949 Dup lnj 0201951 0201951 Dup Inj 0201952 0201952 Dup Inj 0201953 0201953 Dup Inj 0201954 0201954 Dup Inj 0201955 0201955 D_p laj 0201956 0201956 Dup Inj 0201957 0201957 Dup lnj 0201958 0201958 DUPInj 0201959 0201959 Dup Inj Matrix Pup Live_ Pup Livers Pup Liv_ Pup Livers Pup Live_ Pup Live_ Pup Livers Pup Livers Pup Livers Pup Live_ Pup Livers Pup Livu_t Pup Livel_ Pup Live_ Pup LivcrJ Pup Livea'_ Pup Livers Pup Live_ Pup Livers Pup Livca_ Pup Live_ PUpLive_ Pup Livers Pup Liva_ Pup Liv_ Pup Liven Pup Livers Pap Liv_,1 Pup Liveal PUpLivea'_ Pup Livea,J PUpLive_ Pup Live_ Pup Live_ Pup Live_ Pup Live_ Pup Lives Pup Live_s Pup Live_ Pup Liver_ Pup Live_ Pup Liver; Pup Liver_ Pup Livers Pup Livers Pup Llvee; Pup Livers Pup Liven Pup IAv_ Pup Live_ Pup Livers Pup Liven Clleetin Date 5/29/02 5/29/02 5/29/02 5/29/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 Set Number 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 092702A 09270ZA 092702A 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B 092702B PFHS Found (u_/mL) ND ND ND ND ND ND ND ND 21.7 "_ 183 _ 20.5 M 18.9'_ ND ND ND ND ND ND 34.0 _ 30.6_' ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 50.6 M 53.5 M 34.0 _ 37.0 _ 19.6'_ 24.1M ND ND 30.8 "A 34.0 _ '_Although themethodusedallows for alternatesamplevolumesor weightsandall the peakarea responsesare withinthecalibrationcurvelimits,theprecisionforthe 0.1 mLor0.1 g samplesbelow 100 ppbhas not beenvalidated. * DuplicateInjection ND = Not Detected(Arealess thanlowestcalibrationstandardof 0.1 ng/mL) Exygen Research Page 30 of 153 418-028:PAGE G-31 Exygen Study No.: 023-072 Table IX. (eont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor El) 9 19023 GROUP1, Female8" 19023 GROUP !, Female 9 19023 GROUPI, Female9* 19023GROUP I,Fu_alc 10 19023 GROUPL Female 10" 19023GROUP l, Female 11 19023 GROUPL Female 11" 19023 GROUPL F_maalc12 19023 GROUPI, Female 12" 19021 GROUPL Female 10 19021 GROUP I, Female 10" 19041 GROUPI, Male 1 19041 GROUPI, Male 1" 19041 GROUPI, Malc 2 19041 GROUP L Male 2" 19041 GROUPI, Male 3 19041 GROUP I, Male 3* 19041 GROUPI, Male 4 19041 GROUP LMale 4" i 9041 GROUP I, Male 5 19041 GROUP I,Male5* 19041 GROUPL Female8 19041GROUP I,Female 8* 19041 GROUPI, Female9 19041 GROUP L Female 9" 19041 GROUPI, Female 15 19041 GROUPI, Female 15" 19041GROUP I,Formic 16 19041 GROUPI, Female 16" 19041 GROUPI,Female 17 19041GROUP I, Female 17" 19004 GROUPH, Male2 19004GROUP 11,Male 2* 19004 GROUP11,Male 3 19004GROUP ILMale 3" 19004GROUPII, Male 4 19004 GROUPIIM,ale4" 19004 GROUPILMale 5 19004 GROUP1I,Malc 5" 19004 GROUP11 Male 6 19004 GROUP lI, Male 6* 19004GROUP I_ Female 9 19004GROUP H Fuaalc 9* 19004GROUP11 Femalc 11 19004 GROUPH Femalell* 19004GROUP lI Female 15 19004 GROUPII Female 15" 19004GROUPlI Female 16 19004 GROUPII F_nal16" 19004GROUP II Female 17 ..I .9004GROUPI1,Female !7" Exygeu ID 0201960 0201960 Dup Inj 0201961 0201961 Dup Inj 0201962 0201962 Dup haj 0201963 0201963 Duplnj 0201964 0201964 Dup Inj 0201950 0201950 Dup Inj 0201965 0201965 Dup Inj 0201966 0201966 l_p laj 0201967 0201967 Dup Inj 0201968 0201968 Dup Inj 0201969 0201969 Dup Inj 0201970 0201970 DupInj 0201971 0201971Duplaj 0201972 0201972 Dup Inj 0201973 0201973 Dup Inj 0201974 0201974 Duplnj 0202025 0202025 Dup Inj 0202026 0202026 Dup Inj 0202027 0202027 Dup Inj 0202028 0202028 Dup Inj 0202029 0202029 Dup laj 0202030 0202030 Dup laj 0202031 0202031Duplnj 0202032 0202032 Duplnj 0202033 0202033 Dup Inj 0202034 0202034 Dup In_ Matrix Pup Livess Pup Livers Pup Livers Pup Livers Pup Livers Pup Livess Pup Live_ Pup Livers Pup Livens Pup Liv_ Pup Live_ Pup LiveTs Pup Livers Pup Livers Pup Livers P_ Live_ Pup Livers Pup Livess Pup Livers Pup Livess Pup Livers Pup Livers Pup Livers Pup Live_s Pup Livers Pup l..iv_ Pup Livers Pup Livers PUP Liva_ Pup Livers Pup Livers Pup Liven; Pup Liveas Pup Livers Pup Livers Pup Livers Pup Livea's Pup Livers Pup Liv_ Pup Liv_ Pup Livers Pup Livers Pup Livers Pup Livers Pup Live_s PupLiveTs Pup Livers Pup Livers Pup Livers Pup Liver's Pup Livers pup Liven Collection Date 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/29/02 5r29/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31102 5/3 1/02 5/3 1/02 5/31/02 5131/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5131/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 Set Number 0927028 0927028 0927028 0927028 0927028 0927028 0927028 0927028 0927028 092702B 092702BR 0927028R lO0102A 100102A 1O0102A 100102A 100102A 100102A 100102A 100102A 100102A 100102A 100102A lO0102A I00102A IO0102A 100102A 100102A 100102A 100102A 100102A IO0102A 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR 100102AR PFHS Found (u_mL) ND ND ND ND ND RD ND ND ND ND ND ND ND ND 24A _'_ 26.6 '_ 26.0 '_ 24.7 AA ND ND ND ND ND ND 30.6 M 30.8_ ND ND ND ND ND ND 1220 1160 1160 1120 2380 2480 1670 1670 2070 2000 1870 1880 1740 1710 1070 1060 1450 1530 946 983 '_Although themethodused allows for alternatesamplevolumes or weightsandall the peak area responsesarewithinthe calibrationcurve limits, the precisionfor the0.1 mL or 0.1 g samplesbelow 100 ppbhasnotbeen validatecL *Duplicate Injection ND = Not Detected(Arealess thanlowest calibrationstandardof 0.1 ng/mL) Exygen Research Page 31 of 153 418-028:PAGE G-32 Exygen Study No.: 023-072 Table IX. (cont'd). Summary Sponsor ID i9018 GROUP I1, Male 1 19018 GROUP If, Male 1" 19018 GROUP l_ Male 4 19018 GROUP lI, Male 4" 19018 GROUP II, Male 5 19018 GROUP If, Male 5* 19018 GROUP H, Male 6 19018 GROUP If, Male 6* 19018 GROUP If, Male 7 19018 GROUP II. Male 7* ! 901S GROUP II, Femtale 9 19018 GROUP I_ Female 9* 19018 GROUP 11.Female 10 19018 GROUP i1. Female 10" 19018 GROUP 11, Female 11 19018 GROUP !_ Female 11" 19018 GROUP K, Female12 19018 GROUP 11.Female 12" 19018 GROUP II, Female 14 19018 GKOUP II. Female 14" 19026 GROUP H, Male 5 19026 GROLrp 1I. Male 5* 19026 GROUP II, Male 1 19026 GROUP II, Mal i* 19026 GROUP II, Male 3 19026 GROUP II, Male 3* 19026 GROUP If, Male 4 19026 GROUP II, Male 4* 19026 GROUP I/, Male 7 19026 GROUP I1, Male 7" 19026 GROUP If. Female 12 19026 GROUP I_ Female 12" 19026 GROUP If, Female 13 19026 GROUP II, Female 13" 19026 GROUP lI. Female 14 19026 GROUP n, Female 14" 19026 GROUP II. Female 15 19026 GROUP II. Female 15' 19026 GROUP II, Female 18 19026 GROUP II, Female 18 * 19036 GROUP lI, Male 1 19036 GROUP If, Male 1' 19036 GROUP If, Male 2 19036 GROUP iI, Male 2* 19036 GROUP II. Male 4 19036 GROUP II. Male 4* 19036 GROUP II. Male 5 19036 GROUP H, Male 5* 19036 GROUP ii, Male 6 19036 GROUP II. Male 6" 19036 GROUP If. Female 7 19036 GROUP I_ Female 7* Exygen El) ....... 0202050 0202050 Dup Inj 0202051 0202051 Dup lnj 0202052 0202052 Dup lnj 0202053 0202053 Dup haj 0202054 0202054 D_p lnj 0202055 0202055 Dup l.nj 0202056 0202056 Dup lnj 0202057 0202057 Dup Inj 0202058 0202058 Dup lnj 0202059 0202059 Dup Inj 0202063 0202063 Dup Inj 0202060 0202060 Dup Inj 0202061 0202061 Dup Inj 0202062 0202062 Dup Inj 0202064 0202064 Dup Inj 0202065 0202065 Dup Inj 0202066 0202066 Dup lnj 0202067 0202067 Dup Inj 0202068 0202068 Dap Inj 0202069 0202069 Dup Inj 0202070 0202070 Dup Inj 0202071 0202071 D_ lnj 0202072 0202072 Dup Inj 0202073 0202073 Dup Inj 0202074 0202074 Dup lnj 0202075 0202075 Dup In_ of PFHS Residues in Rat Liver Samples Matrix Pup Livers Pup Live_ Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Liv,'rs PUp Live_ Pup Live_ Pup Liven Pup Livers Pup Livers Pep Live_ PUp Live_ Pup Livers Pup Lives Pup Livers Pup Livers PUp Livers PUp Live_ PUp Livers Pup Livers Pup Livers PUp Livers Pup Livers Pup Livers Pup Livers Pup Liv_ Pup Livers PUp Livers Pup Live_ Pup Livers PUp Lives Pup Livea_ Pup Livers Pup Livers Pup Liven PUp Livers Pup Livers Pup Liven PUp Livers PUp Livers Pup Livers PUp Liven Pup Livers Pup Livea's Pup Livers Pup Live_s Pup Livers !_ Liven Collection Date 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/3 1/02 5/31/02 5/3 1/02 5/3 1/02 5/31/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/25/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 . 5/28/02 Set Number 100202A 100202A 100202A I00202A 100202A 100202A 100202A I00202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A 100202A I00202A 100202A I00202A 100202AR 10020ZAR 100202AR IO0202AR 100202AR 100202AR 100202AR 100202AR lO0202AR 100202AR 100202AR IO0202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100202AR 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A 100302A PFHS Found (u_aL) 839 816 788 818 886 872 868 888 523 514 556 596 804 785 649 664 743 751 788 815 937 928 1130 1130 1170 1150 1140 1130 1050 1010 946 956 928 919 1040 1140 1780 1830 1270 1310 746 748 169 174 707 693 965 946 667 649 622 620 * DuplicateInjection ND ffiNot Detected(Area less than lowestcalibrationstandard of 0.1 ng/mL) Exygen Research Page 32 of 153 418-028:PAGE G-33 Exygen Study No.: 023-072 Table IX. (cont'd). Summary Sponsor Emygen ID 19036 GROUP II, Female8 ID 0202076 19036 GROUPlI. Female8* 0202076 Dup lnj 19036 GROUPIf, Female 11 0202077 19036 GROUP 11.Female 11' 020207"/I_p Inj 19036 GROUP1_ Female 12 0202078 19036 GROUP lI, Fmnale 12" 0202078 Dup Inj 19036 GROUP I], Female 13 0202079 19036 GROUP 11.Female 13" 0202079 Dup Inj 19037 GROUP11,Male 6 0202082 19037GROUP 1I.Malc 6* 0202082 Dup Inj 19037 GROUP II, Female 15 0202089 !9037 GROUPII. Female 15* 0202089 Dup lnj 19037 GROUP 1].Male 2 0202080 19037 GROUPI], Mal2" 0202080 Dup lnj 19037 GROUP II, Male 3 0202081 19037 GROUPIf, Male 3* 0202081 Dup Inj 19037GROUP If. Male 7 2083 i983 19037 GROUPII. Male 7* 0202083 Dup Inj 19037 GROUPII. Male 8 0202084 19037 GROUP lI. Male 8" 0202084 Dup Inj 19037 GROUPII, Female10 0202085 19037 GROUPIt, Female 10" 0202085 Dup tnj 19037 GROUPII. Female11 0202086 19037 GROUP lI. Female 1I* 0202086 Dup lnj 19037 GROUPII, Female 13 0202087 19037 GROUP 1I.Female 13" 0202087 Dup lnj 19037 GROUP 11.Female 14 0202088 19037 GROUP II, Female 14' 0202088 Dup Inj 19003 GROUPI11,Male I 0202119 19003 GROUP IlI. Male 1" 0202119 Dup Inj 19003 GROUPIn. Male 3 0202120 19003 GROUP I_ Male 3* 0202120 Dup Inj 19003 GROUPHI, Male 5 0202121 19003 GROUP 111M. ale 5* 0202121 Dup Inj 19003 GROUPIII. MLIe7 0202 !22 19003 GROUP 111M, ale 7" 0202122 Dup lnj 19003 GROUPHI, Male 8 0202123 19003GROUP Ill. Male 8* 0202123 Dup Inj 19003 GROUP III,Female9 0202124 19003GROUP 11I.Female 9" 0202124 Dup lnj 19003GROUP lI1.Female 10 0202125 19003 GROUPIll. Female I0" 0202125 Dup Ymj 19003 GROUP IlI, Female I 1 0202126 19003 GROUP1II,Female 11" 0202126 IXtpl_j 19003GRDUP !1_ Female15 0202127 19003 GROUPI_ Female 15* 0202127 Dup haj 19003GROUP 1_ Female 16 0202128 19003 GROUP!1[, Female 16' 0202128 Dup laj 19007GROUP HI. Mak 1 0202129 19007 GROUP lI_ Male 1" 0202129 Dup Inj 19007 GROUPIll. Male 2 0202130 19007 GROUP_I. Male 2" 0202130 Dup Inj of PFHS Residues in Rat Liver Samples Matrix Pup Live_ Pup Livers Pup Livers Pup Livers Pup Livers PupLivers PupLivers PUpLivers Pup Live_ PupLivers Pup Livers PupLivers PupLiven PupLivers PupLivers Pup Lives Pup Livers Pup Livers Pup Lavm_ Pup Livers Pup Livecs Pup Livers Pup Liven Pup Livew Pup Live_S Pup Livers Pup Livers Pup Liven Pup Liva_ Pup Livers Pup Livers PUpLivers PUpLivm_ PupLivers PupLivers PupLive_ PupLivms Pup Live_ Pup Livers PupLivcrs Pup Livers Pup Livers Pup Livers PUpLivers Pup Liven Pup Livers Pup Livers Pup Livera Pup Lives Pup Livers Pup Liven Pup LiverJ CoUection Date 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30102 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 , Set PFHS Number 100302A Found (uw/mL) 668 100302A 650 100302A 554 100302A 543 100302A 676 I00302A 642 100302A 494 100302A 490 100302A 906 100302A 900 100302A 956 100302A 965 100302AR 1670 100302AR 1670 100302AR 1430 100302AR 1380 100302AR 1280 100302AR 1270 100302AR 1760 100302AR 1770 100302AR 1150 100302AR 1160 100302AR 1300 100302AR 1320 100302AR 1680 100302AR 1670 100302AR 1330 100302AR 1350 100402AR 4640 100402AR 4730 100402AR 6250 100402AR 6320 100402AR 4490 100402AR 4650 100402AR 4540 IU0402AR 100402AR 5820 100402AR 5840 100402AR 4100 100402AR 4170 100402AR 4400 100402AR 4340 100402AR 3270 100402AR 3240 100402AR 4810 100402AR 4920 100402AR 5430 100402AR " 5630 ! 00402AR 3030 100402AR 3090 100402AR 4310 ! 00402AR 4400 * DuplicateInjection ND = Not Deteetexl(Area less thanlowestcelibrationstandardof 0.1 ng/mL) Exygen Research Page 33 of 153 418-028:PAGE G-34 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19007 GROUP HI, Male 3 19007 GROUP IIL Male 3* 19007 GROUP HI,Male 6 19007 GROUP III, Male 6* 19007 GROUP HI, Male 7 19007 GROUP m, Male 7" 19007 GROUP HI, Female 9 19007GROUP II_Female 9* 19007 GROUP HI, Female I 1 19007 GROUP Ill, Female i i" 19007 GROUP Ill, Female 12 19{}07 GROUP HI, Female 12" 19007 GROUP Ill, Female 13 19007 GROUP I]I, Female 13" 19007 GROUP HI, Femal 16 19007 GROUP HI, Female 16" 19008 GROUP HI, Male I 19008GROUP Ill,Male I* 19008 GROUP HI, Male 3 19008 GROUP HI, Male 3" 19008 GROUP HI, Male 4 19008 GROUP IU, Male 4* 19008 GROUP HI, Male 6 19008 GROUP III, Male 6" 19008 GROUP HI, Male 7 19008 GROUP IlL Male 7" 19008 GROUP HI, Female 9 19008 GROUP HI, Female 9* 19008 GROUP HI, Female 10 19008 GROUP IlL Fcnmle 10" 19008 GROUP IIIF,emale II 19008 GROUP HI, Female 11" 19008 GROUP HI, Female 12 19008 GROUP III, F_mal 12" 19008 GROUP HI, Female 13 19008 GROUP HI, Fenud 13" 19013 GROUP HI, Male ! 19013 GROUP HI, Male 1" 19013 GROUP HI,Male 4 19013 GROUP HI, Male 4* 19013 GROUP Ill, Male 5 19013 GROUP HI, Male 5* 19013 GROUP HI, Male 6 19013 GROUP II_ Male 6* 19013 GROUP HI, Male 9 19013 GROUP HI, Male 9* 19013 GROUP HI,Female II 19013 GROUP III, Female 11" 19013 GROUP HI, Female 12 19013 GROUP HI, Female 12" 19013 GROUP HI, Female 13 19013 GROUP HI, Fmnale 13" Exylgea ID 020213 i 0202131 Dup lnj 0202132 0202132 Dup Inj 0202133 0202133 Dup lnj 0202134 0202134 Dup Inj 0202135 0202135 Dup Inj 0202136 0202136 Dup lnj 0202137 0202137 Dup lnj 0202138 0202138 Dup i_ 0202139 0202139 D_p lnj 0202140 0202140 Dup Inj 0202141 0202141 Dup Inj 0202142 0202142 Dup Inj 0202143 0202143 Dup Inj 0202144 0202144 DUP Inj 0202145 0202145 Dup Inj 0202146 0202146 Dup Inj 0202147 0202147 Dup Inj 0202148 0202148 Dup Inj 0202149 0202149 Dup Inj 0202150 0202150 Dup lnj 0202151 0202151 Dup lnj 0202152 0202152 Dup Inj 0202153 0202153 Dup Inj 0202154 0202154 Dup lnj 0202155 0202155 Dup lnj 0202156 0202156 Dup lnj Matrix Pup Live_ Pup Livers Pup Livers Pup Livers Pup Liv_ Pup Livers Pup Live_ Pup Liven_ Pup Livers Pup Livevs Pup Livers Pup Livers PUp Livers PUp Liv_ Pup Live_ Pup Livers Pup Livers Pup Liv_ Pup Livers PUp Livers Pup Livers Pup Livers Pup Liven Pup Live_ Pup Livers Pup Live_s Pup Live_ PUp Livecs Pup Liv_ Pup Livers I_p Live_ Pup LiveTs PUp Livecs Pup Liver_ Pup Livers Pup Livers PUp Live_ Pup Livezs PUp Liva,s PUp Livers Pup Livers Pup Livers Pup Live_ PUp Livers Pup Live_ Pup Livers Pup I.,/ve_ Pup Liv_s Pup Livers Pup Livegs Pup Livers PUp Livers Collection Date ,5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29102 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 Set Number 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402AR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BK 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 100402BR 1004-02BR 100402BR 100402BR 100402BR 100402BR 100402BR PFHS Found (ng/mL) 4450 4370 4430 4600 5630 5720 6340 6330 3850 3820 5230 5130 6630 6870 5720 5690 3920 3810 5080 5070 4090 4070 4810 4750 4690 4380 5190 5050 3070 2770 4120 3760 2790 2480 2440 2340 1590 1790 1950 2050 2030 2090 2420 2480 2180 2280 2130 2150 2000 2010 1680 1780 *DuplicaItnejection ND = NotDetecte(dArealestshanlowesctalibratsitoanndaorfd0.Ing/mL) Exygen Research Page 34 of 153 418-028:PAGE G-35 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Spenser ID 19013 GROUP HI, Ft_ile 14 19013 GROUP HI, Female 14" 19013 GROUP lH, Fcmale 15 19013 GROUP HI, Female 15" 19015 GROUP Ill. Male 1 19015 GROUP I_, Male 1" 19015 GROUP HI, Male 3 19015 GROUP II_ Male 3* 19015 GROUP 1H, Male 6 19015 GROUP HI, Male 6" 19015 GROUP HI, Male 7 19015 GROUP HI, Male 7* 19015 GROUP Ill, Male 8 19015 GROUP HI, Male 8" 19015 GROUP HI, Female 10 19015 GROUP HI, Female 10" 19015 GROUP II_ Female 12 19015 GROUP HI, Female 12" 19015 GROUP III, Female 13 19015 GROUP HI, Female 13" 19015 GROUP HI, Female 14 19015 GROUP l]l, Female 14" 19015 GROUP IH, Female 15 19015 GROUP HI, Female 15" 19005 GROUP IV, Male 1 19005 GROUP IV, Male 1" 19005 GROUP IV, Male 2 19005 GROUP IV, Male 2* 19005 GROUP IV, Male 3 19005 GROUP IV, Male 3* 19005 GROUP IV, Male 5 19005 GROUP IV, Male 5* 19005 GROUP IV, Male 6 19005 GROUP IV, Male 6" 19005 GROUP IV, Female 8 19005 GROUP IV, Pcmale 8* 19005 GROUP IV, l:emale 10 19005 GROUP IV, Fmude 10" 19005 GROUP IV, Female 11 19005 GROUP IV. Female I1" 19005 GROUP IV, Female 12 19005 GROI3P IV, Female 12" 19005 GROUP IV, Ycmalc 15 19005 GROUP IV, Female 15" 19035 GROUP IV, Male I 19035 GROUP IV, Male I * 19035 GROUP IV, Male 2 19035 GROUP IV, Male 2* 19035 GROUP IV, Male 4 19035 GKOUP IV, Mal 4" 19035 GROUP IV, Male 5 Exygem ID 0202157'r' 0202157 Dup Inj 0202158 0202158 Dup Inj 0202159 0202159 Dup _j 0202160 0202160 Dup lnj 0202161 0202161 Dup lnj 0202162 0202162 Dup lnj 0202163 0202163 Dup Inj 0202164 0202164 Dup lnj 0202165 0202165 Dup Inj 0202166 0202166 Duplnj 0202167 0202167 Dup Inj 0202168 0202168 D_p lnj 0202219 0202219 Dup Inj 0202220 0202220 Dup lnj 0202221 0202221 Dup lnj 0202222 0202222 Dup lnj 0202223 0202223 Dup lnj 0202224 0202224 Dup Inj 0202225 0202225 Dup Inj 0202226 0202226 Dup Inj 0202227 0202227 Dup lnj 0202228 0202228 Dup Inj 0202229 0202229 Dup lnj 0202230 0202230 Dup lnj 0202231 0202231 Dup Inj 0202232 19035 GKOUP IV, Male 5" 0202232 I_p h_ Matrix Pup Li_=s Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Liv_z Pup Livers Pup Livezs Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livezs Pup Livers Pup Livm_ Pup Livers Pup l.avers Pup Lira's Pup Livea_ Pup Livers Pup Livers PUp I.avers Pup I..ive_ Pup Livers Pup Livers Pup Livers l_p larch Pup Livl_ Pup Live_s Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers PUp Livers Pup Livers Pap Liven Pup Livers Pup Lives Pup Livc_ PUp Livers PUlpLive_s Pup Livers pup Livers Pup Livers Pup Livect Pup I.iv_s Collection Date 5/29/02 5/29/02 5/29/02 5/29/02 5/31/02 5/31/02 5/3 i/02 5/3 i/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 i/02 5/31/02 5/31/02 Set Number 100402BR 100402BR 100402BR 100402BR 100902AR IO0902AR I00902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 1009(_AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AK 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR 100902AR IO0902AR 100902AR 100902AR 10'd902BR 100902BR 100902BR i 00902BR 100902BR 100902BR 100902BR 100902BR, PFHS Found (a_aL) " 2290 2300 2000 2070 2950 2840 2780 2840 2860 2940 3150 3220 4680 4710 3800 3800 4280 4180 4000 3950 3800 3890 5330 5420 8860 8800 9280 9370 10500 I 1100 8340 9100 11200 10900 11400 11500 12300 12800 i 2600 12800 9010 9100 11800 12000 11500 12000 14500 14500 13700 13600 11300 10800 * DuplicateInjection ND = Not Detected(Area less thanlowest calibrationstandardof 0.1 ng/mL) Exygen Research Page 35 of 153 418-028 :PAGE G-3 6 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor Ex'ygen ID 19035 GROUP IV, Male 6 ID 0202233 19035 GROUP IV, Male 6* 0202233 Dup Inj 19035 GROUP IV, Femalc 8 0202234 19035 GROUP IV, Female 8* 19035 GROUP IV, Female 10 0202234 Dup laj 0202235 19035 GROUP IV, Female 10' 0202235 Dup Inj 19035 GROUP IV, Female I ! 0202236 19035 GROUP IV, Female 11" 0202236 Dup lnj 19035 GROUP IV, Female 12 02027.37 19035 GROUP IV, Female 12" 19035 GROUP IV, Female 13 0202237 Dup lnj 0202238 19035 GROUP IV, Female 13" 0202238 Dup lnj 19039 GROUP IV, Male 1 0202239 19039 GROUP IV, Male 1 * 19039 GROUP IV, Male 2 0202239 Dup Inj 0202240 19039 GROUP IV, Male 2* 0202240 Dup Inj 19039 GKOUP IV, Male 3 19039 GROUP IV, Mal 3* 0202241 0202241 Dup Inj 19039 GROUP IV, Male 4 ! 9039 GROUP IV, Male a* 0202242 0202242 Dup lnj 19039 GROUP IV, Male 5 0202243 19039 GROUP IV, Male 5* 19039 GROUP IV, Female 8 0202243 Dup lnj 0202244 19039 GROUP IV, Female 8* 0202244 Dup lnj 19039 GROUP IV, Female 11 0202245 19039 GROUP IV, Female 11" 19039 GROUP IV, Female 12 0202245 Dup Inj 0202246 19039 GROUP IV, Female 12" 0202246 Dup Inj 19039 GROUP IV, Female 14 0202247 19039 GROUP IV, Female 14" 0202247 Dup Inj 19039 GROUP IV, Female 15 0202248 19039 GROUP IV, Female 15" 0202248 Dup Inj 19040 GROUP IV, Male 2 0202249 19040 GROUP IV, Male 2* 0202249 Dup Inj 19040 GROUP IV, Male 3 0202250 19040 GROUP IV, Male 3* 19040 GROUP IV, Male 4 0202250 Dup Inj 0202251 19040 GROUP IV, Male 4* 19040 GROUP IV, Male 5 0202251 Dup lnj 0202252 19040 GROUP IV, Male 5" 0202252 Dup inj 19040 GROUP IV, Male 7 19040 GROUP IV, Male 7" 0202253 0202253 Dup Inj 19040 GROUP IV, Femah: 11 0202254 19040 GROUP IV, F_al 11" 19040 GROUP IV, Female 12 0202254 Dup Inj 0202255 19040 GROUP IV, Female 12" 19040 GROUP IV, Female 13 0202255 Dup Inj 0202256 19040 GRDUP IV, Fcamlc 13" 0202256 Dup lnj 19040 GROUP IV, Female 15 0202257 19040 GROUP IV, Female 15" 0202257 Dup lnj 19040 GROUP IV, Female 16 0202258 19040 GROUP IV, Female 16" 0202258 Dup Ini Matrix Pup Liven Pup Live_ Pup Liv_ Pup Livers Pup Liven Pup Liven Pup Live_ Pup Livels Pup IAvcn Pup Live_ Pup Livers Pup Lives Pup Livers Pup Live_ Pup Liven Pup Livers Pup Livers Pup Liva_ Pup Livers Pup Liven Pup Liven Pup Liva= Pup Liven Pup Livers Pup Live_ Pup Livez_ Pup Livers Pup Live_ Pup Livers Pup Livexs Pup Live_s Pup Livers Pup Livers Pup Liver_ Pup Lives Pup Livers Pup Livm Pup Liven Pup Livers Pup Livers Pup Livm Pup Liven Pup Livers Pup Livm Pup Live_'_ Pup Liver_ Pup Liven Pup Liven Pup Livers Pup Liven Pup Live_ Pup Liven Collectmn Date 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/I)2 5/31/02 5/31/02 5/31102 5/31/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/29/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 Set Number 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 10O902BK 100902BR 100902BR 100902BR 100902BR IO0902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR !00902BR i00902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR 100902BR IUI002AR 101002AR i 01002AR 101002AR 101002AR 101002AR 101002A.R. 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR 101002A 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR PFHS Fmmd (n_tml.,) 10600 10500 12000 13300 1{3000 10400 18500 18400 12600 12000 17300 17300 5320 5200 6160 6310 5300 5150 5870 6130 6500 6430 4910 5010 3510 3710 4550 4620 4310 4180 5110 4920 6480 7690 I 1500 11200 7540 7750 13000 12200 ! 1100 II100 12100 11200 10600 10700 14100 14000 12400 12300 14900 15700 *Duplicate Injection ND = Not Detected(Arealess thanlowestcalibrationstandardof 0.1 ng/mL) Exygen Research Page 36 of 153 418-028:PAGE G-37 , Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor Exygea ID 19045 GROUP IV, Male 1 El} 0202259 19045 GROUP IV, Male 1* 0202259 Dup lnj 19045 GROUP IV. Male 2 19045 GROUP IV. Male 2* 19045 GROUP IV, Male 4 19045 GROUP IV, Male 4* 19045 GROUP IV, Made 5 0202260 0202260 Dup lnj 0202261 0202261 Dup Inj 0202262 19045 GROUP IV. Male: 5* 19045 GROUP IV, Male 6 0202262 Dup Inj 0202263 19045 GROUP IV, Male 6* 0202263 Dup Inj 19045 GROUP IV. Female 8 0202264 19045 GROUP IV, Female 8* 19045 GROUP IV. Female 9 0202264 Dup lnj 0202265 19045 GROUP IV. Female 9* 0202265 DUP Inj 19045 GROUP IV. Female 10 0202266 19045 GROUP IV, Fanal 10" 0202266 Dup lnj 19045 GROUP IV, Female 11 19045 GROUP IV, Female 1I* 19045 GROUP IV. Female 14 0202267 0202267 Dup Inj 0202268 19045 GROUP IV. Female 14* 0202268 Dup laj 19006 GROUP V, Male I 0202320 19006 GROUT V, Male 1" 0202320 Dup Inj 19006 GROUP V, Male 2 19006 GROUP V. Male 2" 0202321 0202321Dup Inj 19006 GROUP V, Male 3 0202322 19006 GROUP V, Male 3* 19006 GROUP V, Male 4 0202322 Dup I_ 0202323 19006 GROUP V, Male 4" 19006 GROUP V. Male 5 0202323 Dup lnj 0202324 19006 GROUP V. Male 5* 0202324 Dup Inj 19006 GROUP V, Female 8 19006 GROUP V, Female 8* 0202325 0202325 Dup Inj 19006 GROUP V. F_m] 10 0202326 19006 GROUP V, Female 10" 19006 GROUP V, Female 11 0202326 DUP Inj 0202327 19006 GROUP V, Female 11" 19006 GROUP V, Fc_le 13 0202327 Dup Inj 0202328 19006 GROUP V, Female 13" 0202328 Dup Inj 19006 GROUP V, Female 18 19006 GROUP V. Female 18* 0202329 0202329 Dup lnj 19011 GROUP V, Male 2 19011 GROUP V, Male 2" 0202330 0202330 Dup lnj 19011 GROUP V. Male 3 0202331 19011 GROUP V. Male 3* 19011 GROUP V, Male 4 0202331 DUP Inj 0202332 19011 GROUP V, Male 4* 0202332 Dup Inj 19011 GROUP V, Male 5 0202333 19011 GROUP V, Male 5" 19011 GROUP V, Male 6 0202333 Dup lnj 0202334 19011 GROUP V, Male 6" 0202334 Dup lnj 19011 GROUP V, Female 9 0202335 19011 GROUP Vr Fcmud9" 0202335 Dup laj Matrix Pup layers Pup Liwr5 Pup IAv_s Pup Livers Pup Live_ Pup Live_ Pup Live_s Pup Livers Pup Live_ Pup Livc_ Pup Livers Pup Livers Pup Livers Pup IAv_ Pup Livers Pup Live1 Pup L_v_ Pup IAvc_ Pup Lwe_ Pup IAvem Pup Lives Pup IAven Pup lAve_ Pup IAve_ Pup Live_ Pup Li-c_s Pup lAven PUp Liven Pup Livers Pup Liven Pup IAve_ Pup Livers Pup Lave_ Pup Ltve_ Pup Livers Pup Live_ Pup Lives Pup Livers Pup Livers PUp Livevz Pup IAv_ Pup Liv_ Pup Livcs_ Pup Live_ Pup IAven Pup Livers Pup Livers Pup Live_m Pup Livew Pup Live_ Pup IAvcra Pup Live_ Collection Date 528/02 528/02 5/28/02 528/02 5/28102 5/28102 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 528/02 528/02 528/02 5/28/02 5/28/02 5/28/02 5/28/02 5/28/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30102 5/30/02 5/30102 5/30/02 5/30/02 5/30/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 Set N,amber 10100ZAR 101002AR 101002AR 101002AR 101002AR 101002AR 101002AR I 01002AR I 01002AR 101002AR 101002AR 101002AR 101002AR 101002AP. I01002AR I01002.AR IDI002AX 101002AR 101002AR 101002AR 101002BK 101002BR 101002BR 101002BR 101002BR 101002BR 101002BR 101002BR I 01002BR 101002BR 101902BR 101002BR 101002BR 101002BR 101002BR 101002BR 101002BR 101002BR 101002BR 101D02BR 101002BR 101002BR 101002BR I01002BR I 01002BR 101002BR 101002BR 101002BR 101002BR 101002BR 101002BR 101002BR 'PFHS Foa_ad (a_nL) 6200 6210 4830 4900 7820 7510 5700 5750 4840 4970 7350 7350 6510 6740 8350 8010 6520 6380 6030 5870 13500 13700 16500 16700 25800 26100 30100 29400 30200 29800 23300 22500 17700 16400 25000 25000 16700 16500 20700 20200 19200 20000 18700 19900 19300 19000 18601) 19300 14000 14100 20800 21100 *DuplicatIenjection ND = NotDetecte(dArealessthanlowestcalibratisotnandarodf0.1ng/mL) Exygen Research Page37 of 153 418-028:PAGE G-38 Exygen Study No.: 023-072 Table IX. (cont'd). Summary of PITHS Residues in Rat Liver Samples Spenser n) 19011 GROUP V, Female 10 19011 GROUP V, Female 10" 19011 GROUP V, Female 11 19011 GROUP V, Fea_al 11" 19011 GROUP V, Female 12 19011 GROUP V, Female 12" 19011 GROUP V, Female 13 1901 i GROUP V, Female 13" 19020 GROUP V, Male 2 19020 GROUP V, Male 2* 19020 GROUP V, Male 4 19020 GROUP V, Male 4" 19020 GROUP V, Male 5 19020 GROUP V, Male 5" 19020 GROUP V, Male 7 19020 GP.OUP V, Male 7* 19020 GROUP V, Male 8 19020 GROUP V, Male 8* 19020 GROUP V, Female 9 19020 GROUP V, Female 9" 19020 GROUP V, Female 10 19020 GROUP V, F_aalc 10" 19020 GROUP V, Female 11 19020 GROUP V, Female i I* 19020 GROUP V, Female 12 19020 GROUP V, Fe_aal 12" 19020 GROUP V, Femal 13 19020 GROUP V, Female 13" 19922 GROUP V, Male I 19022 GROUP V, Male 1" 19022 GROUP V, Male 4 19022 GROUP V, Mal 4" 19022 GROUP V, Male 5 1907.2 GROUP V. Male 5" 19022 GROUP V, Male 7 19022 GROUP V. Male 7* 19022 GROUP V, Male 8 19022 GROUP V, Mal 8" 19022 GROUP V, Female 11 19022 GROUP "_, Female 11" 19022 GROUP V, Fm'aale12 19022 GROUP V, Femal e 12' 19022 GROUP V, Female 13 19022 GROUP V, Female 13" 19022 GROUP V, Female 14 19022 GROUP V, Female 14" 19022 GROUP V, F_tl 15 19022 GROUP V, Female 15" 19025 GROUP V, Male 2 19025 GROUP V. Male 2* 19025 GROUP V, Male 7 Erygea n) 0202336 0202336 Dup Inj 0202337 0202337 Dup lnj 0202338 0202338 DUP lnj 0202339 0202339 Dup lnj 0202340 0202340 Dup lnj 0202341 0202341 Dup Lnj 0202342 0202342 Dup Inj 0202343 0202343 D_p l.nj 0202344 0202344 Dup Inj 02021345 0202345 Dup Inj 0202346 0202346 Dup laj 0202347 0202347 Dup lnj 0202348 0202348 Dup Inj 0202349 0202349 Dup Inj 0202350 0202350 Dup Inj 0202351 0202351 Dup Inj 0202352 0202352 Dup laj 0202353 0202353 Dup lnj 0202354 0202354 Dup Inj 0202355 0202355 Dup lnj 0202356 020235613_ap laj 0202357 0202357 Duphaj 0202358 0202358 D.p Inj 0202359 0202359 D_p Inj 020236 i 0202361 Dup lnj 0202362 Matrix Pap Live_s Pup Livers Pup Livers Pup Livers Pup Live_rs Pup Live_ Pup Livers Pup Livers Pup Livers Pup Livers Pup Live_s Pup Live_ Pup Live_ PUP Live_ Pap Livers _ Livers Pup Livers Pup Livers Pat)Livers Pup Lives PUp Livers Pup Lave_ Pup Livers Pup Livers pup Livers Pup Livea's Pup Livers Pup Livers P_ Live_ Pup Live_ Pup Live_s Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livc_ Pup Livers PUp Live_ Pup Live_ Pup Livers Pup Livers Pup Livers Pup Livers Pup Liver_ Pup Livers Pup Livers PUp Lives PUp Live_s Pup Livers Collection Date 5/31/92 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/3 1/02 5/3 1/1)2 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/3 1/02 5/3 1/02 5/3 1/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/31/02 5/31/02 5/31/02 5/3 1/02 5/31/02 5/30/02 5/30/02 5/30/02 5/30/02 Set PFHS Numb.e.r..., Foun__(u_mL) 101002BRR 18700 101002BILR 20000 101002BRR 23500 101002BRR 101002BRR 22400 21900 101002BRR 22500 101002BRR 21000 101002BRR I 01102AR 21400 I 1800 101102AR 101102AR i 1300 9460 IOI 102A.R 9190 1011_ 8130 101102AR 8510 101102AR 9280 101102AR 101102AR 9050 8750 101102AR 8750 101102AR 14800 101102AR 101102AR 14300 12200 101102AR 101102AR 12600 14400 101102AR 14300 101102AR 15600 I01102AR 101102AR 15300 20900 101102AR 101102AR 21300 ! 1100 101102AR 101102AR 11000 12700 101102AR 1011 02AR 12400 12500 101102AR 12800 101 ! 02AR 10i 102AR I 1800 11600 101102AR 10n02AR ! 4400 14300 101102AR 13700 101102AR 101102AR 14100 17400 101102AR 101102AK 17500 12200 101102AR 101102AR i 1600 15700 101102AR 15300 101102AR 14100 101102AR I 01402AR 14200 21700 101402AR 21900 101402AR 18600 *DuplicateInjection ND = Not Detected (Area less than lowest calibrationstandardof 0.1 ng/mL) Ex'ygenResearch Page 38 of 153 418-028:PAGE G-39 Exygen Study No.: 023-072 Table IX. (eont'd). Summary of PFHS Residues in Rat Liver Samples Sponsor ID 19025 GROUP V, Male 8 19025 GROUP V, Male _* 19025 GROUP V, Male 13 19025 GROUP V, Male 13" 19025 GROUP V, Female 14 19025 GROUP V, Female 14" 19025 GROUP V, Female 15 19025 GROUP V, Female 15" 19025 GROUP V, Female 18 19025 GROUP V, Female 18" 19025 GROUP V, Female 19 19025 GROUP V, Female 19" 19025 GROUP V, Female 20 19025 GROUP V, Female 20* 19025 GROUP V, Male I 19025 GROUP V, Male 1" Exygen ID 0202363 0202363 Dup laj 0202364 0202364 DUP haj 02021365 0202365 Dup Inj 0202366 0202366 Dup Inj 0202367 0202367 Dup lnj 0202368 0202368 Dup Inj 0202369 0202369 Dup lnj 0202360 0202360 Dup Irti Matrix Pup Liven Pup Live_ Pup Liven Pup Livers Pup Livers Pup Livers Pup Livers Pup Livers Pup Livex_ Pup Live_ Pup Livers Pup Livers Pup Livers Pup Livers Pup Live_ Pu_ Liver_ Collection Date 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30102 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 5/30/02 .., 5/30/02 Set Nug_r 101402A1_' 101402AK 101402AR 101402AR 101402AK 101402AR 101402AR 101402AK 10140ZAR 101402AR 101402AR 10140ZAR 101402AR 101402AR 101402ARR 101402ARR PFHS Found (nl/mL) 21400 21_00 18300 19400 14700 15300 26400 26500 19800 19500 18900 18800 16800 17600 13300 14400 *DuplicateInjection ND = Not Detected(Area less thanlowestcahq_rationstandardof 0.1 ng/mL) Exygen Research @ i Page 39 of 153 418-028:PAGE G-40 Exygen StudyNo.: 023-072 Table X. Summary of PFHS Residues in Dosing Solutions Sample l[_e_tion B-418-028-A (07Jim.02) 0 mg/mL 1 of 4 B-4 |8-028-A (07Jun.02) 0 mg/mL 1 of 4 B-418-028-B (24.May.02) 0.03 mg/mL 1 of 4 B-418-028-B (24.May.02) 0.03 mg/mL 1 of 4 B-418-028-C (24.May.02) 0.1 mg/mL I of 4 B-418-028-C (24.May.02) 0.1 mg/mL 1 of4 B-418-028-D (07Jun.02) 0.3 mg/mL 1of 4 B-418-028-D (07Jun.02) 0.3 mg/mL i of 4 B-418-028-E (24,May.02) I mg/mL I of 4 B-418-028-E ('24.May.02) 1 mg/ml. 1 of 4 Bulk TA/S Sample (09Jun.02) Bulk TA/S Sample (09Jun,02_ Set Number 110702C 110702C 110702C 110702C 110702C 110702C 110702C 110702C 110702CR 110702CR 110702C 110702C PFHS Known Concentration 0 0 30000 30000 100000 100000 300000 300000 1000000 1000000 100000 100000 PFEIS (nl[/mL_ Found (nE/mL) ND ND 26500 27300 88900 91000 268000 278000 1010000 1000000 125000 1220_3 PFHS Recovery _%_) 88 91 89 91 89 93 10! 10{3 125 122 ND= NotDetected(Arealessthanlowestcalibratiosntandarodf0.1ng/mL) ExygcnResearch Page40 of 153 418-028:PAGE G-41 Exygen Study No.: 023-072 Table XI. Summary of PFHS Residues in Stability Samples 3M m E02-0S 14-38611 E02-0514-38611" E02-0514-38612 I_U2-0514-38612" E02-0514-38613 E02-0514-38613" E02-0514-38614 E02-0514-38614" E02-0514-38615 E02-0514-38615" E02-0514-38616 E02-0514-38616" E02-0514-38617 Eft2-0514-38617" E02-0514-3861B E02-0514-38618" E02-0514-38619 E02-0514-3_,619" E02-0514-38620 E02-0514-38620" _n m 0203923 0203923 De _iaj 0203924 0203924 Dy _Inj 0203925 0203925 De _laj 0203926 0203926 De _ Inj 0203927 0203927 De _ luj 07_392_ 0203928 De _ laj 0203929 0203929 De ) l_j 0203930 0203930 De ) laj 0203931 0203931 D_ _ htj 0203932 0203932 Dup Ini Sami_ ,, m..:,,_o. B-418-028 A 129 Mi 02) 0 mf./mL 1_4 B-418-028 A (29 Mar 02) 0 mg/mL 1 of 4 13-418-028A (29 Mar 02) 0 mg/mL2 of 4 B-418-U28 A (29 Mar 02) 0 mg_L 2 of 4 B-418-028 B (29 Mar 02) 0.03 mg/mL ! of 4 15-418-028B (29 Mar 02) 0.03 ml_aL 1 of 4 B.418-028 B (29 Mar 02) 0.03 ms/mL 2 of 4 B-418-028 B (29 Mar 02) 0.03 mf/mL 2 of 4 B-418-028 C (29 Mar 02) 0.l mg/mL i of 4 B-418-028 e (29 Mz 02) 0.1 mg_aL I of 4 B-418-028 C (29 M_ 02) 0.1 mg/mL 2 of 4 B-418-028 C (29 Mar 02) 0.1 mg/mL2 of 4 1_-418-028D (29 Mar 02) 0.3 mll/mL 1 of 4 B-AIS-028 D (29 Mar 02) 0+3ms/mL 1 of 4 B--418_28 D (29 Mar 02) 0.3 tag/mL 2 of 4 B-A18-028 D (29 Mar 02) 0.3 mg/laL 2 of 4 B-4184128 E (29 Mar (r2)I mS/mL l of 4 B-418-028 E f'Z9Mw02) I ml_mLI of 4 B-418-028 E (29 Mar 02) 1 mS/ml, 2 of 4 B-418-028 E _9 Mar 02) I m_/mL2 of 4 Set .,LS',_.. 110502A 110502A 110502A !10502A 110502A 110502A 110502A 1105O2A 110502A 110$0ZA 110502A 110502A 110502A l]tlS02A 110502A 110502A 110702AR 110702/_ 110702AR 110702AR PFI]S Kwwl CoK. q,I/-_.) 0 0 0 0 30000 30000 30000 30000 100000 100000 I00000 I00000 3110000 300000 300000 300000 1000000 10000_ 10G0000 1000000 I'FHS PFBS Found Reeevery (,,1_t4 e_) NU ND ND ND 29700 99 29600 99 29700 99 29500 98 122000 122 117000 117 100000 100 100000 100 308000 103 316000 105 326000 109 324000 108 910000 91 919000 92 1030000 103 1010000 101 *DuplicaItnejection ND =NotDetecte(dArealestshanlowesctalibratsitoanndaorfd0.lng/mL) Exygen Research Page 41 of 153 418-028 :PAGE G-42 Exygen Study No.: 023-072 Table XII. Summary of PFHS Residues in Homogeneity Samples 3M Exygea Sample Set El) El) De_J_ttea Number E02.-0514-38621 0203933 B-4I 8-028-._ (29Mar02) 0 mg/mL 1 of 12T I 10702A E02-0514-38621" 0203933Duplaj 13-418-028-A (29Ma:02) 0 mg/mL 1 of 12T 110702A E02-0514-38622 O203934 B-418-028-A (29Mat02) 0 mg/mL 2 of 12T 110702A Eff2-0514-38622. 0203934Dep Inj B-418-028-A (29Mar02) 0 mg/mL 2 of 12T 11r0702A E02-OSI4-3S623 0203935 B-418-028-A (29Mar02) 0 mg/mL 5 of 12M 110702A E02-.0514-38623" 0203935Dup lnj B-418-028-A (29Mar02) 0 mg/mL 5 of 12M 110702A E02.-0514-38624 0203936 B-418-028-A (29Mar02) 0 mg/mL 6 of 12M I 1070ZA E02-0514-38624" 020393613_ lnj B-418-028-A (29Mar02) 0 mg/mL 6 of 12M 110702A EO2-OSI4-3S625 0203937 B-418-028-A (29Mar02) 0 ms/mL 9 of 12B !10702A E02-0514-38625' 0203937Dup Inj B-418-028-A (29Mar02) 0 ms/mL 9 of 12B !10702A E02-0514-38626 0203938 B-418-028-A (29Mat02) 0 mg/mL 10 of 12B i 10702A Eff2-0514-38626" 11203938Duplnj B-418-028-A (29Mm92) 0 mg/mL 10 of 12B 110702A E02-0514-38627 0203939 B-418-028-B (29Mar02) 0.03 _ l ofl2T 110702A E02-0514-38627" 0203939Dup Inj B-418-028-B (29Mm02) 0.03 mg/mL 1 of 12T 110702A E02..051_38628 0203940 B-418-028-B (29Max02) 0.03 mg/mL 2 of 12T 110702A E02-0514-38628" 02039,40Duplnj B-418-028-B(29Mar02)0.03mgh_.2of12T 110702A E02-0514-38629 0203941 B-418-028-B ('29Mar02) 0.03 ms/mL 5 of 12M i I0702A E02-0514-38629" 0203941Dup In.j B-418-028oB (29Mat02) 0.03 mg/mL 5 ofl2M 110702A E02-0514-38630 0203942 B-418-028-B (29Maz02) 0.03 miP'mL6 of 12M 110702A E02-0514-38630" 0203942Dup Inj B-418-028-B ('29Mat02) 0.03 mg/mL 6 of 12M I10702A E02-0514-38631 0203943 B-418-028-B (29Mar02) 0.03 mg/mL 9 of 12B 110702A Eff2-0514-38631" 0203943D_ lnj B-418-028-B (29Ma.-02)0.03 mg/mL 9 of 1213 110702A E02-0514-38632 0203944 B-418-028-B (291Vim02)0.03 mg/mL 10 of 12B 110702A E02.0514-38632" 0203944Duplnj B-418-028-B(29MmO2)0.03mg/mL10of12B |10702A E02-0514-3S633 0203945 B-418-028-C (7.9Max02)0.1 mg/mL 1 of 123" 11ff'/02A E02-0514..38633" 0203945Duplnj B-418-028-C (29Mat02) 0.1 mg/mL 1 of 12T 110702A E02-0514-38634 0203946 B-418-028-C (29Mar02)0.1 mshnl., 2 of 12T 110702A E02-0514-38634" 0203946Duplnj B-418-0284= (29Mar02) 0.1 mghnL 2 of 12T 110702A E02-0514-38635 0203947 B-4 !8-028-C (29Mar02) 0.1 mg/mL 5 of 12M 110702AR E02-0514-38635' 0203947DupInj B-418-028-C (29Mar02) 0.1 mg/mL 5 of 12M i 10702AR E02-0514-38636 0203948 B-4! 8-028-C (29Mar02) 0.1 mg/mL 6 of 12M 110702A E02-0514-38636" 0203948DupInj B-.418-0284= (29Mar02) 0.1 mg/mL 6 of 12M 110702A E02-0514-38637 0203949 B-418-0284= (29Mar02) 0.1 ms/mL 9 of 12B 110702A E02-0514-38637. 0203949DupInj B-418-028-C (29Mar02) 0.1 mg/mL 9 of 12B 110702A E02-05|4-3863_ 0203950 B-4 | 8-0284= (29Mm02) 0.1 mg/mL |0 of 12B | 10702AR E02-0514-38638" 0203950Duplnj B-418-028-C(29Mar02)0.1mg/mL10of12B 110702AR E02-0514-38639 0203951 B-418-028-D (29Mar02) 0.3 ms/n_ I of 12T !10702A E02-0514-38639' 0203951Dupinj B-418-028-D (29Mar02) 0.3mg/mL i of 12T 110702A E02-0514-38640 0203952 B-418-.028-D (29Mat02) 0.3 mghnL2 of 12T 110702A E02-0514-38640. 0203952Dep lnj B-418-028-D (29Mar02) 0.3 mg/mL2 of 12T 110702A E02-0514-38641 0203953 B-418-028-D (29Maz02) 0.3 mg/mL 5 of 12M 110702B E02-0514-38641" 0203953_ tnj B-418-028-D (29Mar02) 0.3 mg/mL 5 of 12M 110702B E02-0514-38642 0203954 B-418-028-D (29Mar02) 0.3 mghnL 6 of 12M 110702B F.02-0514-38642" 0203954Dep Inj B-418-028-D (29Mart72)0.3 mg/mL 6 of 12M 110702B EID.-O$14..38643 0203955 B-418-028-D (29Mm'ff2)0.3 mg/mL 9 of 12B I 10702B E02-0514-38643" 0203955I)uplnj B-418-028-D (29Mar02) 0.3 mg/mL 9 of 12B l 10702B E02-0514-38644 0203956 B-418-028-D (29Mar02) 0.3 _ 10 of 12B 110702B E02-0514-30644* 0203956Duplnj B-418-028-D(29MmO2)0.3mg/mL10of12B i10702B E02-0514-38645 0203957 B-418-028-E (29Mar02) I mg/mL l ofl2T 110702B E02-0514-38645" 0203957_ lnj B-418-028-E (29Mm02) i mg/mL l of 12T 110702B E02-0514-38646 0203958 B-418-028-E (29Ma.'02) 1 ms/rid.,2 of 12T 110702B E02-0514-38646" 0203958Duplnj B-418-028-E (29Mar02) 1 mg/mL2 of 12T ! 10702B E02-0514-38647 0203959 B-4I 8-028-E (29Mar02) 1 mg/mL 5 of 12M 110702B B02-0514-38647" 0203959DupInj B-418-028-E (29Mat02) I mg/mI..5 of 12M 110702B E02-0514-38M8 0203960 B-418-028-E (29Mar02) 1 mg/mL 6 of 12M 110702B E02-0514-38648' 0203960Dup haj B-418-028-E (29Mar02) 1 mlghnL6 of 12M 110702B Eft2-0514-38649 0203961 B-418-028-E (29Mar02) I mg/mL 9 of 12B 110702B E02-0514-38649" 0203961Duplnj B-418-028-E (29Mar02) 1 mg/mL 9 of 12B 110702B E02-0514-38650 0203962 B-418-028-E (29Mar02) I mg/mL 10 ofl2B 110702B E02-0514-38650" 0203962Dup lnj B-418-028-E (29Mar02) I mg/mL 10 of 12B 110702B 'PFHS Known Cone. (a_mL) 0 0 0 0 0 0 0 0 0 0 0 0 30000 30000 30{)0<) 30000 30000 30000 30000 30000 30000 30000 30000 30000 100000 100000 100000 100000 100000 100000 100000 100000 100000 100000 100000 100000 300000 300000 300000 300000 300000 300000 300000 300000 300000 300000 300000 300000 1000000 10000013 1000000 1000000 1000000 1000000 1000000 1000000 11300000 1000000 1000000 1000000 PFIIS Foand _a_/mL) ND ND ND ND ND ND ND biD ND ND ND ND 24.40O 23700 26900 25800 25500 26100 25800 25800 24200 244O0 24800 24500 75400 73800 76300 74700 93300 95100 69800 72300 84400 83700 95700 92800 270000 268000 270000 279000 254000 266OO0 241000 239000 255000 250000 280000 293000 1020000 1090000 987000 980000 1160000 1170000 1040000 994O00 1090000 110000(3 1110000 1070000 PFHS Recovery (%) 81 79 90 86 85 87 S6 86 81 81 83 82 75 74 76 7_ 93 95 70 72 84 84 96 93 90 89 90 93 85 89 80 S0 S5 83 93 98 102 109 99 98 116 117 104 99 109 tl0 111 107 * Duplicate Injection ND = Not Detected (Area less than lowest calibration standard of 0.1 ng/mL) Exygen Research Page 42 of 153 418-028:PAGE G-43 Exygen Study No.: 023-072 FIGURES Exygen Research Page 43 of 153 418-028:PAGE G-44 Exygen Study No.: 023-072 Figure 1. Typical Calibration Curve for PFHS Compound 1 name: PFHS Coefficientof Determination0: .999484 Calibrationcurve:2633.68 x+ 44.4695 Responsetype: ExternalStd, Area Curvetype:Linear.Origin:Exclude.Weighting:l/x, Axistrans:None 1.32e4- Response 0 ..... ng/mL 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 ExygenResearch Page44 of 153 418-028 :PAGE G-45 Exygen Study No.: 023-072 Figure 2. Chromatogram PFHS Representing a 0.1 ng/mL standard for C082002-6. 0.1 ng/mL Standard 091g02B-2012 Sm {Iv_, 27,3) 100- I.e4_ _9 214el_2002 10:34:30 LC4M$/MS dr? MRM Of 1 _ ES399>80 4.21e3 Anm 1.00 2.00 3.00 4.00 5.00 6.00 L 7.00 8.00 9.00 10.00 11.00 t2.00 Exygen Research Page 45 of 153 418-028:PAGE G-46 , Exygen Study No.: 023-072 Figure 3. Chromatogram Representing a Control Rat Plasma Sample for PFHS (Exygen ID 0202913 Control, Data Set: 091602B) 0202913 Control 0916029-207 1 Sm (J_. 2x3) o.17 3.M_ I Y g,m W._M_rrS 17'-Sep-2002 22:52:59 k4FUvol f 1 Channel c 399 > 8_ 3.00 5.45 6.60 9.22 55 10.21 I _ t0.75 1.00 01 73 2.00 3.00 3.91 4.00 4m 4.93 S.O0 5.U6 6.00 O,T 77 LO0 8.00 I ___ 9.00 10.00 nFTRI11"56 11.00 1ZOO Exygen Research Page46 of 153 418-028:PAGE G-47 Exygen StudyNo.: 023-072 Figure 4. Chromatogram Representing a Control for PFHS (Exygen ID: 0202987 Control, 091902B) Rat Serum Data Set: Sample 0919(3G_-200_ Sm (f_. 2x3) 1oo-1 Iva_4 of 1 _ lO.18 ES3_09 80 289 020--2t987 Control A r! 1.19 1.00 tt_ M.2"41 _i_J3_._L_4.30 4.1022 2.00 3.00 4.00 5.00 5,72 6,00 7.06 7.00 IL_ &814 9.29 11,34 21-Sep.2DL0.2C_M0S9/:k0_7:34_7 10.46 11.11_ 8.00 9.00 10.00 11,00 12.00 Exygen Research Page47 of 153 418-028:PAGE G-48 Exygen Study No.: 023-072 Figure 5. Chromatogram Representing a Control Rat Liver Sample for PFHS, (Exygen ID: 0202877 Control, Data Set: 100202A) 0202877 Control 10Q20_A-20S8m (Mn,27..3) 100- 03-0t-2002 08"17.:19 t O/MS/MS #7 MRMof 1C_annelES- &3z 399>80 S3_ 2.00e3 % 1.00 2.00 3.00 4.00 5.00 15.00 7.00 8.00 9.00 10.00 11.90 1ZOO Exygen Research Page 48 of 153 418-028:PAGE G-49 Exygen Study No.: 023-072 Figure 6. Chromatogram Representing Control Rat Plasma Sample Fortified with 10 ppb of PFFIS (Exygen ID: 0202913 Spk A, Data Set: 091602B) 0202913 Spk A, 10 ppb 0916028-208 Sm (Mn, 2x3) 1QO 17-Selv2002 23:14:44 LC_SlMS 87 _ of 1 C_anne_E,_ 9.oe 399 >8( _7" B.25e2 Area 1.00 2.00 3.00 4.00 15.00 6.00 7.00 8.00 g.o0 10.00 11.00 12.00 Exygen Research Page49 of 153 418-028:PAGE G-50 Exygen StudyNo.: 023-072 Figure 7. Chromatogram Representing Control Rat Serum Sample Fortified with 10 ppb ofPFHS (Exygen ID: 0202987 Spk A, Data Set: 091902B) 0202987 Spk A, 10 ppb 091902B-2009 Sm (Mn, 2x3) 100_ It.14 54g - 21-6ep-2002 09".29:14 Lr.dMSIMS dr7 t,m_l of 1 _ ES- 399 =_80 'I.D4e4 Area _.00 2.00 3.00 4.00 5.00 6,00 7.00 B.O0 9,00 10.00 11.00 12.00 Exygen Research Page 50 of 153 418 028:PAGE G51 Exygen Study No: 023-072 Figure 8. Chromatogram Representing Control Rat Liver Sample Fortified with 10 ppb of PFHS (Exygen ID: 0202877 Spk A, Data Set: 100202A) 0202877SI_ & 10PI_ 100202A.,.209Sm (fAn, 2x3) 100 t 0_-2002 08:.3_S'/ LC_IS/MS#7 MRM 04'1 Cl'_nnel ES- s30 399.'80 2_- 1.A0Ererea4 1.00 2.00 3.00 4.00 5.00 (LO0 7.00 8.00 9.00 10.00 11.00 12.00 Exygen Research Page51 of 153 418-028:PAGE G-52 Exygen Study No.: 023-072 Figure 9. Chromatogram Representing Rat Plasma Sample Analyzed for PFHS (Exygen El): 0201816, Sponsor ID: 19176 GROUP I, Data Set: 091602B) 02011116 0916029-212Sm(Mn.2x3) 100- 9.07 2772- 11-,._q)-2002 00:41:24 L.C_SAUS dr7 MRIoI rI Cha3m9_9>E8S02.16o4 Anla % 1.00 2.00 3.00 4,00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 Exygen Research Page52 of 153 418-028:PAGE G-53 Exygen StudyNo.: 023-072 Figure 10. Chromatogram Representing Rat Serum Sample Analyzed for PFHS, DF=10 (Exygen ID: 0201863, Sponsor ID: 19079 GROUP H, Data Set: 091902BR) 0201863, OFBt0 0919021SR"308Sm (Mn. 2x3) 100 &a0 24_" 24-,,Sep-Z002 02:50:15 LC/MSJ'MS #,r/ MRM of 1 Chactn_ ES399'> 1_0 3.91 e4 Area 1,nn 2.00 3.00 4.00 5.00 6.00 7.010 8.00 9.00 10.00 11.100 12.00 Exygen Research Page53 of 153 418-028:PAGE G-54 Exygen StudyNo.: 023-072 Figure 11. Chromatogram Representing Rat Liver Sample Analyzed for PFHS (Exygen ID: 0202050, Sponsor ID: 19018 GROUP II Male 1, Data Set: 100202A) 0202050 100- 03.._)Q2 10.'00:52 LC_M,_qNSIIr"V 0,3 3_ > 8C) 21_ 1.19e5 Area _t L 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 g.00 10.00 11.00 12.00 Exygen Research Page 54 of 153 418-028:PAGE G-55 Exygen Study No.: 023-072 APPENDIX A Study Protocol 418-028 (Exygen Study No. 023-072) and Amendments, Deviation and Note to File Exygen Research Page 55 of 153 418-028:PAGE G-56 Exygen Study No.: 023-072 _ _ _tt _ Hm_M 1_O4_ "- r,_ als7_TJo ra_ ass9.jasa7 ARGUSRESEARCH Char/a_ Labormor/es O/mm_ end_ Sen_ PROTOCOL 418-028 SPONSOR'S STUDY I_3MBER: T-7706.1 STUDY TITLE: Oral (Garage) Combined RepeatedDose Toxicity Study of T-7706 with the Reirmdu_on/Developmontal ToxicixyScreening Test PURPOSE: _srmc FAEI_TY: STUDY DIREL'rOI_ SPONSOR: The purpose of thisstudy is to provideinformationonthe possible health hazards that may remJltfrom repeated exposureof CrI:CD_SD)IGS BR VAF/Plu_ male and femalerainto a t_ substancebeginning before u_aabi_ion, throushma_8 end u_n_nalngfor at least 42 days (male rats)or tl_uughpmluritiou tmu_day 21 of lactatiou(female rm). This repeated dose studyinnorpomes a _pred_tlon/devciopmcat_ tmficity screm_g test thatcan be used to provideinitial in_conmfionon pos_'ble effects on male and femalerepmd.clive lxncmmmace(e.g. gonadal functima,mating behavior,conception,development of the mnceptm end pamu-ition). The studyxko places m=phaslsou ueuroloi;i_al_ts as a specific cndpoinl andshould identifythencurotoxic potential of a test substance,which may warrantfurt_ in-depthinvcstigation. Because of the _-'lectivityof the and_inm and the shortdurationof the study, the _ test will not provideevidence for defim_e claims of no re_ducticm/developmmtal effects. Inpanicute:, it offers o..dylimited means of detecthlg postnatal manifestationsofl_ttal _ or_ that maybe induceddrainSpostnatalezposere. Argus Research 9O5Sbeehy Drive. _u_lding A Homham,Penmytveni_ 19044-1297 Telephone: ('215)443-8710 Tlefax: (215) 443-8587 Raymond G. York, Ph.D. DABT As_ate Director of Reseat_ Email: raymond.york_riv=.com Address as cited above for Testing Facility 3M CorporateToxicolow 3M Cent_, Buildi_S 220-2F.-02 SL Paul. Minnesot5a5144-1000 Exygen Research Page 56 of 153 418-028:PAGE G-57 Exygen Study No.: 023-072 Pmto_141&_.8 P,ge2 "_ STUDY .MONITOR_ John Butenhofl_Ph.D. DABT,CIH 3M_ Toxi_toSy 3M Medical Telephon_. (651) 733-1962 Telefax: (65t)733-1773 _mil: jlbut_off@mmm._ REGULATORY CITATIONS: Orgmisationfor Economic Ca-operation endDevelopment (1996). OECD Guide_for Fang of_ic_t/s. Sectima4, No. 422: CombinedRepcaledDose Toxicity Study with the _epmental ToxiciStmy_mugTesta. dopta2i2March199_. Orga_ueifcomr_ Co-ope_oanndDmlopmm(199ZT)h.eReviuOz_t"D Pnncip_ofGoodLaboneor_yacticm[_97)186/Fkmt]. U_. Food andDrug Administnt_ Good LabmatoryPracticeResu]a_om; Final Rulc. 21CFR Part$8. $apmae_Me inistry of Health andWelfare (1997). Good Laboratory Pracace ,gtandardfor ,_fety Studies awDru_, MHW OrdinanceNumber21, March26, 1997. tt_CUt_TORYC_O_a'L_CE: TI_ studywill be conducted in mmpllanc,e with the GoadLaborelotyPt-tctice(GLP) rcgul_am cited above. All changes or _ of thispmlocol _tli be documented,signed by the Study Dire_or theSponsor, dated mad_ with theprotocol. The TeC.ingFacilitQyusalit_y Unit (QAU)willauditthe protocol, the rawdam and the report,and willinspecctrilical _ of those pro'floraof the study conductedat the "Ie_in8 Facility in eccordanccwith the StandardOperating_ of theTesting Facility. The finalreportwill include a compliance_ateme_ signed by the StudyDirector that",han:port aczuratelyrr,flet_ the rawdata obtainedduringtheperfon_anceof the study and thatall applkable GLP regulalimaswere followed in the conduct of the study. Should significant devi_om fromGLP reBu]ationosccure,achwillbed_sc_'oienddetaitlog,mh_ with how tim deviationmight affect the quality or integrityof the study. Should anypot't/onof the studybe _nd_ted by a subconlnctor or by the Sponsor, the Study Dkectorwill m_m tha_a qualifiedPrincipalIn-veatigatoirs identifiedby the facility conducting thatpe_tionofthc study. The QAU forrhlsfacilRywill conduct _ phase impec_ end auditre_= results endrcim_ forthatgady lmrtiou aemrdiag to the _OPe of that fac_. Such _ phase inspection reportsandreport auditswill be submittcdby thz fadlity to the Prln_alI_tigator mdtheStudyDim:t_. Thedateoefthe_ mdreport subnd_ons will be incorporau_ into a QAU Statementgencmtedby that tk_h'tyendprovided Exygen Research Page57 of 153 418-028:PAGE G-58 Exygen Study No.: 023-072 h_3 "_ to the Testing Facih'tyforinclusion in the final report. Inaddition, this fitcilitywill providea sm_mem of GLPcompliance, _. ducribed shove, signed by the Principal]nve_i_or for inchudo_ in _e final repor_ SCHEMATIC OF STUDY DESIGN AND STUDY S.C"HEDULE; See ATrACHMENT I to tb_ proWcol. TEST SUBSTANCE A_']) VEHICLE: IdeulifJ_tloa: Test Subetsn_ 1"-7706 [P_uoroh_mc Sulfu_e PotassiumSalt (PFI_] Lot idcmific_on will be docummmi in the rawdata. The Sponsorwill p_ovideto theTcs_ug Fac/lity documentationor cctti_ation of the identity, eomp_Uon, mefl_xolf ryutl_, _l_h andactiv/_/pur/ty of the u_ subm_c_ This documentationwill be includedin the final report. AqueouOs _5"m/,'ooxymethlyceUu(l_ose (mediumviscosityp)repareudsingreverse o,ano_s membrane processeddeionized wate_(P.O. de/ouizedwatef_ Lot ide_i_.at/on and Supplierwill be documentedin the ntw data. Neither the Sponsornor the Study Directoris awere of any potential ccmtmninm_ likely to be presem in the vehich that would int_esc wlth the resultsof this study. _ no analyses otherthanthoee mentioned in this protocolwill bc conducted. Safety Pru:au/lons: Gloves, dust-_A-fil_:red mask,zppropri_ eye protection and mffonn/lab coat to be worndmmg _'mu_ion prepm_i_ and douse. The MaterialSafety Data Sheet (MSDS) is anachedtoth_protoco(lATrA_ 2). s_._: BulkTostSubmnce: Bulk Vehicle _: PrepareTdestSubmnce andVehicle Formulations: Room tanperatm'e. Room temperstu_. Refrigerated(20C w 80C). .-. All teatsubatance ahipmentstlmuid be addremmdto the _n of Julian Gulbinaki,Mamger of FormulationLtbonaory, atthe previously cited Testing Fscility addresaandteleplmm nuanbe_, Exygen Research Page 58 of 153 418-028:PAGE G-59 Exygen Study No.: 023-072 Proto_418-028 _4 "_ Shitmummldlouki i_ludc im_m2,_m _ slm'agc _ m_tsl_pl_g r.._,.o_ should be labeled appmpriztely. Therecipient thouldbe notified in advance oftbip_ent. FORMULATION: Frequency of Preparation: Fozmulafiom(suspe_dc_) will be preparedweekly at the Testing Facility. Detailed preparationprocedureswill be allachedto this protocol (ATTACHMENT3). Mmlluat/_t._tl_ Tbe test sul_q=a:e will be eomidet_ 100% pureforthe purposeof dosaSe _. Tes/i_ Facil/ty l_erve Sanmles: The Testing FL'ility will reservea umapleof esrh lot of bulk teat substance (appmximateb/1 g) and bulk vehicle compone_ (apprc_a_tely I g or 5 mL) u_:! duringtbe _tmm of the i_a_y. Samples will be stored underthe previously cited conditien_ of requiredanalysmwill be pmvlded to the Teetin8 Pa_h'ty forinclasinn in the study report. Samples additionalto these _ below maybe taken ifdaemed Mcemry dartingthe course ofthe stay. AdditionalanalyJe_ ifreq_tin_ will be _ted bypmtm_l a_aendme_ Balk Test Smlmmee Samo_. A sample of apprmdmately1 g of thetest mblaance, will be takenon the lest day ofla_atment end iem (ambient condition,) to: Principal_tm': Li_ 3M BnvirmnnentalTeclmolosy andSafety Servic_ Building 2-3E-09 St. Paul, Minnemta 55133-3331 Teleplume: (651) 778-$$68 Telefax: (651) 778-6176 laclemen@mn_.mm The recipient will be notified in advtu_ ofumpl ehipmlmt. Exygen Research Page 59 of 153 418-028:PAGE G-60 Exygen Study No.: 023-072 Amdvses of l_"emweMFermalaflem: Co_lzafion and Hom_tv: Pmtlx_ 418--028 _5 Concentrationa_l homogeneity ofthc prcparai formulationswill be verified duringthe coune of this study. Quadmpllcate samples (2 mL cach)will be taken from tbe top, middleand bettom of each cmr.ealx_oa on the firstday of preparation. Two samples fi_m each quadruplicateset will be sbippat for analysis;the rcmainin8samples waqlbe retained at the Testing Facility u backupsamples. Quadrtzplic_tesamples will be taken from each conc_mtmtiouon the last day of preparation.Two samples flora each quadruplicateset will be skipped foranal_is; the re_0_is_insgamples will be n;tsina:la backupsmnplcs. Backup samples will be ston,'dunda' the previousclyitedconditionasnddiscardeadttheTestin8FacilityupontherequesotftheSponsor. Stab_. Stability of the percparedfommlafionswill be documcntcdduring tiffsstudy. Two sets of dnplicatesamples (2 mL each) fromeach concentrationwill be taken on the firstday of ptcparafion. One umple ofeach dupllcate set will be shipped on the day ofpreparalion. These samples will be analyzedat the following time points: as seen after prepamlionu pomu'blend tm days afterthe first anal3_c Tbe ranaininll umples will be retainedattbe Tes_ug Facility u backuprdlnlple&_ Slllnplmwill be lltored Ilndorthe previously cited condifim_sand discardedat theTurin 8 Faefl/tyuponthe requestof the Sponsor. Samplesto be tn_yzcd will be shipped(refrigerated)to: prin_psl_nvestigatoLrm. Ckmm 3M Envimnmemt_Technology andSafety Services Bm'Iding2-3E-09 St. Paul, _ 55133-3331 Telephone: (651) 778-5568 Telefax: (651) 778-6176 Email: laclemen_znmm.mm The recipimt will be notified in advauce of sample shipment, DISPOSrrlON: Preparedformulationswill be discardedatthe Tcsting Facility. All remainingbulk test subetaneewill be returnedto: Dan Hake= 3M EHSR- Auto & Chem Cap _. 3M Centcr,Building23_1B-10 st. PaulM, trmeso5mSit4-1000 Telepimae: (651) 733-2392 Exygen Research Page 60 of 153 418-028 :PAGE G-61 Exygen Study No.: 023-072 "I_,ST SYSTEM: Spet_e_tra_.mad Resmu for Seleclloa: Protoco4l 11141_ rise6 The CrI-CD_SD)IGS BR VAF/PIu_ ratwas _ as the Test System bccaus: I) it is one mammalianspecies ar.,e_ted foruse in toxicity studies amdit has been widely used O_zoughou_ indmstr2y),thistraionfrathasbeend_ tobesensititvoereproductainvde developmenttaolxinsa;nd3)historidcaatlaandexperienecxeisatttheTestinFgacili_t3y_ lmfial populal_onacclimated: Populationselected for mainstudy:. 100 male and 100 virgin female rats. 75 male and 75 _ finnale rats(_5pe_ sex per Populafionaelected for toxicokln_ study: lSmaland 15 fcmalerats (threepef sex per dosagegroup). Male rats will be ortlm_ to weigh from 275 g to 300 g e_ch sa receipt, atwhich time _ _ be expected to be at leut 60 daya of age. Female rats will be oxdczedto weigh from 200 g to 225 g each at receipt, atwhie.htime they will be eapected to be at least 56 daya of a_. Actusl body A weights will be recorded the dayII1_ receipt lindwill be documclxtedin the raw data. The weight ranges will be inf,luded ixtthe final report. At stuffyinil_liou, the weight vuriafiouof the r_ will not exceed :_.20%of the mcnoweight of each ae_. Sex" Both male and female ratswill be evahmted. Cl_rlez River Laboratories,Inc. The ntts will be slaippedin ffltsred cartonsby air fieight and/ortrack from Chadm _v_ Laboratories,Inc., to the TestingFacility. .ldemflfleatioa: Pals m,epcrmmae_y idmatificdus_ag Mond sctf-pi=cingcartags (Gey Band andTag Co., Ino., No. MSPT20101). Maloax_tfuma_ rats azc amigncd_ nombcrs at ra_ipt _nd givm uniquepamanent id_iflcation m=nberewhen assigned to the study before adminiatration of thefirst doea_. Pups will not be individuallyidentified durin8 lactation; all parametenwill be evaluated in terms of the lira. Exygen Research Page 61 of 153 418-028:PAGE G-62 Exygen Study No.: 023-072 " ANIMAL HUSBANDRY: 418..1128 Pqp '7 Ali ca_ sizos and hcusing co_J_ns me in compliancc wilh th=Uuidefov the Cart _ U_eof Laboratory Animals _). Argus_ is an AAA_C-w, crodit_l f_'i_ty. Fo genm'atio_ratswill Ix: _ndiv/duallyhouscd in stainless s_=l wire-bottomed cages except duringthe cohabitationandpostpartumperiods. During mlmbim_on,each pairof ratswill be housed in lJ_ male rat'scsLc,. Beginning no lat_ thanday20 of presumed gl_a_on, Fo gmm'afimxfemale ratswill be individuallyhoused in nestingboxes. Eaahdam anddelivered litterwill be homed in a common mining box during thepompm'Uupneriod. max=-_(bcd-o'cob_w) inbeprovided. Bcddins will bc charted as often u ncccsma_m keep thea'_m_I"dry msdclcsn. Analyses fmposm'blocontaminationarcconductedsemi-mmuallyanddo_maent_in theraw data. Room Air. Temnermtwre and gudditv: --- 'l'_hmoimarloom is_y _l_Hedwithatl_sttm _ l_ah"ourof 100%frea_th tl_htut IxmaInmcdtl_ 99.97%HEPA filt_.Rcmm tmqxa'atmw_ill_ _ at 64F to 79F (18(2to 26C) andmonitor_ constantly. Roomhmnidity will also be monitored comtantlyand maintainedat30% to 70%. klr_t: A_ automaticaclxlmyl_ll_1l2-l_m"light:12-hourdarkfluo_c_t light cyr.wlicllbe Eachdarkpm'iodwill bzg_ at 1900 hoursF._T. Thefight cyclmaybe adjustedby theStudy Directoro designee ffd_nncd r_mury to accommoda_ scheduled laboratory azfivifim. Any such adjustme_ will be docmne_cd in the rawdata. Ramwill be given CertifiedRodent Diet_ _D02 0'MI NutritionIntonational) availablead i/b_mmfromindividual feeders. Feed wilblet-_mn_xtlheevemingpriorto the scheduled uczifice,. w_ter: W_ will be availabalde//hirafmromindividualbottlm mscbcd to the cag_ _ from an automaticwatmi,ng _x_m systmn. All wau:rwill be frmnt local source andpas_ througha r_m'se osmosismcmbnme befo_ un. Chlorine wiUbe addea_othe Imu:emed w'atcru a bacte:ios_ processed waur is expec_ to muta_nuo mm,ema 1.2 ppm r.hlor_ ,,, flu_tim= of anal3mis.Wateris mmly'z_[monthly for pinto'hie bacleriale_'taminata'on_ twice ammallyfor pom'blechemical cmatmnimfion. Exygen Research Page 62 of 153 418-028:PAGE G-63 Exygen Study No.: 023-072 P,mtom4Il&..02S Page8 c_.qmmimm_: Neither tbe Spcmsornor tbe Study Directoris awareof anypotentialcontaminants likely to be prese_inthece_fiediti,nthedrinkinwgaterorinthenestinmgateriaaht Icy=Isthat would int_fm'weith_ r_mltosfthisstudy. Thcrcforn_o,analyses oth_ than those rvutincly performedby the fecd mkoplie=or those mcnfioncdin thimprotocolwill bc conducted. DAY _LrMBKRINGSYSTEM: Gestationday0 is definedas theday spetmatvzoa m observedin a smearo_ the vagin_ conUmmmd/o_a copulatoryplus observed/n _. Thedny of birthmdesignated lactationday 0 (pmtpm'uanday 0) in tl_ Health Ei_cts Test Guidelines- Repmd,,.._on and Fertility Effects (Office of Prevention,Pesticides and Toxic Submnc_ 870.3800, August, 1998) and in the OECD Guidelineforthe Testing of Chemicals CombinedRctmm_ Do_ ToxicityStudywitthhe.Rc_m:lu_on/Dm_lopmmtaTl oxicity Scm_mingTeat(Sectio4n,No.422,22March 1966)T.hissamedayisck=ignat_e*dyl postpartum(day I of lactation) in the StandardOperatingProceduresof the Testing Facility. Tlmmglmuttldpsrotocotlb,edayofbirtwhillImd_4pmt_ daytIm_tmx (d_tIyof l_taliomnd) allmb_mt al_ oftl_F1 gc_a_fiornammaddaysoft_ l_tafiopn_od will tedmmnin_md care_dt_agy. RANDOMIZATION AND COHABITATION: Uponarrivarla,twsill bc assigned to individual housing on thebasis of computer-generated randomunila. Durinllm a_.limaficmperiodof at lea_ five days, male tonifemale ralswill be sele_ed for studyon the basis of phyJ/c_l _ and body weights reunded during acclimmion. The ratswill be usigned to dosage Broupsbasodoa computer-gcacratcd(weight- mitred)nndmniz_mp_rocedun=. Withineada dma_ group, mnse_ ov_:r will be used to arraignramm cohabitatie_, one ratper f:emalerat. Thecohabitatinnpm'iodwill con_tofamaximum ofl4days. Femalerats with q_mamzoa observed in a I=nearof thc vaginalcontents and/ora copulatoryplug observed #n_ttu will be cor.sideredto be ztday 0 oflmmmmd Sest_ion md assigned ta _ housing. Female n_ not mated within the first _ days of cohabitationwill be m_isncd tltemate male rats thathave mated (same dmage group)andwill remainin cohabitationfora maximm_ off.am addilional day=. Day I of i_'tatlo_ _) is defined u the day of birthandis also the Rmtday on whic_ all pupsina litterare individnally weishad (ImPbody weights will be recordedalter all pupsin a liU= _e delivered md grnomed by 1hedam). LittewrislnlotbeoAIeddurintghels_aaion period,becausreandomselection of pups for .,-. culling mmldresult in potentialbiases in pupviabilities and bedy weight gains ov_ _s _ Exygen Research Page 63 of 153 418-028:PAGE G-64 Exygen Study No.: 023-072, iKo,lo_ 41g-0211 l_,t9 "" Withineach doling=81xlup,cail_culive 0rdcf will be _ to Iluign the first l 0 milleand the I0 fomalrats to a fimctional observationalbattery _OB) and motor activity msmsmcnatb, lood s_nplc collcct/cnafor clinical cbemislry ancthcmatolc_ (CCAH), and histological evalm_m, On day22 poslparlxu_ a tlble ofrmdom unitswill I_ used w select five malo _ fi_ _e pupsper litt_ forblood sample and liv_ coll_lioa; these pups win only be sel_-tc_lfrom the tm damssedn_d for POB, motor activity, CCAH andhistolosie_l m'ahnslion. ADMINISTRATION: Route and lhmsoa for Choice: Thereal (pvage) mutc was selo:_d fc_use because: I) in compmqsonw/th the clictarymutc, the exact dosa_ csmbe accuratr.lyszbn/nistcreds;nd 2) it is one oftbe poss/ble mutm fix cnvirmnnon_ Method and Frmuenc_. Ikeages will be edjusted daily forbody we/ghtchanges aad given atappmximm_ythe same lime eachday. The first day of dmage is desisnated as day I ofstudy. Maler_ will be given the test _stanee oace daily beginning 14 days before a cohabitation period thatcon=ishlof a m=_amm 14 dayL Dotage will contimmthroug]lth=daybefor= sacrifice,after eomplelion of the oohab/tat/onp=r/ockaltera minimumof 42 da_ of admietmaam_ Fmude ratswfl/be giv_l the t_t imltmlce once daflybeginning 14 days befo_ acohabilalion periodthat cAmsiataof a maxinnnnof 14 day_ Dosagewill coatinne throughthe daybefore &clmdaledsacrifice (day 21 of lactation). Ra41enalefor l)ma_e Seleetim: l:_a_ wea_ sele_l by the Sponmrbasedoe inevious m_diesmndue_ with _ _ mbmmcn, ud_a_ into eccouat pore'hi=_ in _idvity be_n_= prolmm _ad nonla_tent rtt_ The hish_ dmall= will be expe:ted to umse texi effects bet not mertality or obviousauff=-ing. The de_cedin8 sequenceof the lower dmase level_ wt'llbe aelectedf0rtbe perpose of dememtratin8 a_y dmage-retaXedrespense,with r.oadverseeffects expected atthe lowest level. / A. Exygen Research Page 64 of 153 418-028:PAGE G-65 Exygen Study No.: 023-072 Dont_ L_ Coac_lanttlo_ mad Velum: Pmmc_ 4111-.1)_ ralp: 1o nemSe dL,m Dm*W Cmmm, M,n' Vu_,m I 15 +.3 b 0 0 l0 0,418.038--AlOey,.MmYb.ylar) 1I 15 + 3 t 0.3 In I$ + _ I fV I $ + Sb j V I-5 .,jb 10 0JD'J 0.1 0.3 I 10 ID4 1.8.413_O(12ey.ldJ.Ymr) 10 B-4.1 _.)d,'_t_ y_) I0 B-4.I 8-4]_8-1_Dty.Mmuh.Ycm-} l0 _ t11.026-lB(l:_.Mo_'.yar) _. T"tiam_m_indmtdV_minmupmbe'mmupdaddmoqd1, 0a0m_p?uwre_lnftSmminiimamrlim_a__edoJqe_ rumpmle,_ecm_ ANALYSES AND _S - lro G_TION: Yitbi]K_ - Male tnd Female Rats: All Pcriods: At least twice daily. ClinieJd Observations Lad/or Oeuerxl Annem'wBee- Male mtd Femal e Rats: A,c_ Pu/od: Wce_ly. Doag_ Period: "-- Daily bcfon_ do88ge. On tho flint day of dosage. _ ol_n_tttio_l will be _ tt _x_ti_tt_dy hourly [ntm'vals fur the flint four hours and at the etal of tl_ normawl odan8day.Subsequepnotstdosagoebm-v_ons willbcrccontcdat_atuva_danned_ bythe Study _r or dmiSncc afar dctamin_'on of pcsk toxicologic efi_cct_ Maternal Behavior:. Days I, 5, 8, 15 and22 p_tpartum. Observed behavior ngmded daffy. obsay_ons may bc recofdat more fzequmgly tlan c/ted above, if dcaned appropriatc byth_ Stt_ Director and/or Study Moaitor. Da_ coU_t_t for rats usis_ed to t_0xic.okm_c eampb:collcc_o,, will m)t be summarizedor Detailed Cthtfcai Obseryndons -Mtle 8nd F.emsle Rats: Once bcfom tiac first dosagc madat lcmt once weekly _, derailed clinical observations will bc conducted for all realmad fanale rats. These obsermdions will be m_ie outsid_ the cage in a standard arc_ at the samc time each day of conduct. Effort will be mad_ to insure that variations in the test u_itior_ 8re _ and that obsm-vations are comiucted by obs_ycrs _-_ unawareoftxeatmeutgmt_. SisnJ noted should include, but not be limited to: dumgeai_el6n, fur. eyes, mucous membrmm, occurrmce oframmlom and cxactiom mad maonomic nctivity (c._, lacrim_on, piloerec_m, pupil size, unusutl rmpiratory pattern_ Chanscs in gait, posture Exygen Researeh Page 65 of 153 418-028:PAGE G-66 Exygen Study No.: 023-072 Pmtec_ 4| g-028 P',t_ 11 md _ to handlin8 u well as lye premace of clouic or ionlc movezn_s, sl_rotypic behavior(e.g., cxceuive vooming, repetitive circling), difficult orprolonl_i parturitionor behavi_ (e.g., self..mun'Iationw, alking backw, ards) should also be record_ Body Wetelm - M_ Bd Fcnmle ibm: Acclim_on Pct'i_: Weekly. Dosage Period: Daily. Sacrifice: Tmnimd weight. Feed Consumtion Values - Male I_U (recordedendtabulated): Dosage P_od: Wt=ldy. ]_eedConsumntloa Values - Female _ (recordedandtabulated): Dosag_ Pcriod: W_,lr.lyto cohabitation. Gestatiou Period: Days 0, 7, 10, 12, 15, 18, 20 and25 (if necessary). PosttuutumPcriod: Days !, 5, 8 and 15. Feed consumptionnot tabulatedaft= day 15 postpm.tumw, hen it is e_pect_ thatpups will begin _oconstant matenud feed. Feed Co_umntiu Vslues - MIdemad Female lbb: Feed cmuumplion ve2uesmaybe rcconiedmo_ frcqumdy thancitcd above if it is nec_ m replenishthe feed. Duringcohabitation,whc_ two rats occupy thc same cage with one feedjar, rcplcnishmc_ of thc feedjarswill be documented. Individualvalues will not be recorded or tabulated. Tozieokiuetle Simmle Collection: On day 14 and 42 of study,blood sample8 (approximately! mL sch)will be collected from each male ratassigned to thetoxicokinetic scruplecollccfion portionoftho study (3 pet'_). Onday 14 ofstudy and day 21 ofprcstnned gestation,blood samples (approximatelyI mL each) will be collectcd from each fcmaleratassigned to the toxicokinetic sample collection portionof thc study (3 pcr group). Samples will be collected priorto douge on day 14of study. Thetime of cach blood collection will be rccordedin the rawdata. Exygen Research Page 66 of 153 418-028:PAGE G-67 Exygen Study No.: 023-072 Pmm_41&412g Blood will be mllected fax_mthe mbital sinuL If neceuary, blood may be collected from an alternatedte_ if m, foe alternaterite will be decumm_ in tbe ntw data.) Thz samples will bc _ in_ EDTA_ (pmple top)tubm md sp_/n z centrifuge. Tbe reaulling scram will be la'ansfen'edinto polypmpylene tubu labeledwith the protocol nnmbeL Sl_etamrstudy number,animal manber,u=_ gmep numbe_,dmage level, dayof study,colleeti_n interval,date of colh_on, _e, gm=-afio_endatmaiF:conditions. All samplcs will be immediatelyfrozen az dry ice end maintainedfrozen (.@70*C)until shipment foranaly_ia All= the last blood sample collection, ratswill be taeaiticed 8nd samples oftbe liver wall be collected for maly_ SidDehtl lutrnetleas: Samplm_ be analyzedwill be ahippedmadry ic_ _: PrincipalInvcstigstur:LisaClem= 3M EnvironmentaTlechnologyandSafetyServices Building 2-3E-09 St Paul, Minnesota 55133-3331 Telephone: (651) 778-5568 Telc_x: (651) 778-6176 Email: laalemcnOnmma.cem ,..,-. The recipimt will be notified in advanceof sample shipme_ _4Jtreus_ and Mltiu: Estrouscycling will be evaluatedby exmninationof vasinal cytology belong& with the day afteIrbefirsetdminis_ andthentm_1spermatozaoxaeobserveidna l_neaorffoev_ cow,tentsend/orz copulatoryplug i8 obu=vcd _ s_u dsuingthe cohabitationpcdod. Camreu-Seetioahte- Toxicokiutic Study: On day 21 of presumedgestalio_ blood andliv_ t_nples will be OlleOtedfi'om 811female rats dedgnated furmxicokinm'csample _lleetieL Blood rumple8will be collected fromthe rsts as pt_-vioualydeac_dbed.Afo_ temifice_ the liver of eachratwillbe exeimi and the liverweight will b recmded. The median riverlobe will be frozen andstoeed -_20%-')entil shlpmentforpoulble amlysi,, The kdwea will be removed fi'omlh ateaamandblood samplm will be cellected from each fetus via de.fit.'on. Blood will be placed intolabet, lmO|edper litter, allowed to clot mui spunin a _eifese; tbo _estdan8_ will be U-_sfmed into labeled polypmpylene tubes. All samples w_l be immediatelyfiuzm en dryice andmaiatainedfiezee (_70"C) until _nmt for Exygen Research Page 67 of 153 418-028:PAGE G-68 Exygen Study No.: 023-072 Protoco4l18-02,1 hgc 13 "" Theliver _ mcdafetus will be collected, pooled _ litter ted placed m_ labeled tubes. "rhc m_plm will be fzezm aad stored (_<-20"C)until sbipmcnt for ana/ysis. Samples will be shipped on ch_ ice to Lisa Clcmm at tbc previously cited acldmss. Nataral Dtllvev'_r: Female rats will bc evaluated foc. Adverse Clinical Signs Obsaved DuringP'm_lioa. Duration of C-cst_on (day 0 ofprcsumcd gestationto the timc thc firstpup is obscrvcd). Litter Sizc (deemedas all pupsdclivcrcd). Pup Vi_b_ty E Birth. Fanctioasd Observational Bsm_c. On one oec_on during the courseof the study,s finmtioealobsca'vatlonalbatty (FOB)cs4)will be cct_kmed on 10 male md 10 f_mmlchas l_r _q_, For male ha& this ameum_ will be conductedshortly be.forescheduled sacrificc, but priorto blood sample coUnction for _" hematology and clinical _ cv_ns. Fcnudcrats slmuldbe tested duri_ thelactation paiod, shortlybefore scheduled nm,ifiee. TheFOB, to be condactod by tmobeezm_ unawm'eof the group_signmeut of the rat, wa'llas_s the following pazmneters: 1. _oa. salivation, palpebral closure, pmmin_e ofthc eye,t_pilla_ reaction to light,piloct_tioa, rcspiratimam, adurinationand dcfecation (tm_aomicfu_eo_). 2. Scmccimotm"reSlmnamto visual, auditory,tactile andpainful stimuli (reactivity and sai_ivity)o 3. Reactions to handling andbehavior in the opeu field (excitability). 4. Gait pattern in the open field, severity of gait abnemmlities, air righting reaction, and landing foot splay (gait and sensorimotorcoordination). _, Fo_imb m_l_ _ sla'aag_ 6. Almmmal clinical signs includingbut not limitcd to comrulsious, trancn and other umamal behavior,hypoton_ orhypc_mia, maaciation, dchydratiort, ..- mdcempt sppearame and deposits aromd the eye_ n_e or mouth. Exygen Research Page 68 of 153 418-028:PAGE G-69 Exygen Study No.: 023-072 l_otoco1418.o28 Pip 14 _'_ Evidence of the ability ofthia batm'y to detectthe effects of posiu'vecentroIsubstanceswill be providal frcs_ Fac_y Pmiavc Connol Dam). Dm win a_o be provided to docummat intembscrvcr _ if more tlma me obser_r is involvat in tbe uming. Mptor Aettvitv Te_ Motor activity will be mmlualzdon 10 male and 10 female ratspar grouponce during_ _ of the study. This assesammt will be condut_edshortly beforeacbeduled sam.ifice,but priorto i blood sample coUcctio_, i Themovemc_sof .ar_ratwiUbemomita_dbya paaive infrm'ede_,ac__ _ a slainkms-slcelwh'e-_ rage (40.6 x 25.4 x 17.8 cma).Eachtest I_mion will be 1.5 horn in durationwith the numberofmoveme_ and time spcm in movema_ tabulatedat e_h five- mimue_ The appars/uswill monitor t rack ofup to 32 oagcsand scmors dreamscech scssima,with ceda rattmml in the mine loomion mathe rick eu:_ tm smaiom. Oroupawialbe comm4nlmced m'_ testin_ smsimmemdca_. Dm will be providcd to danonsU_tc am aac zst systan is cweble of daec_ m in activityproduc_ by positive omla'ol aubmn_ CrestingFacility Pceitive _1 Data). _TOLOGY AND CLIIqlCAL _: .--. At tl_ed sacrifice, the _ (fm_i) mllgnzd to ham_tology tnd limc._lchemialzy(HkC_ manplecollection will be _ from the inf=-ioryam cava following _ by carbondioxide emphyxiaficmA. Iq,mxlme_y 5 mL of blood will be collected and Woct_ed as d_crt_:d bclow. Dctmmimtiom mklifionalto thosc dcacn_o_lbelow may be _ tf tbe known pmpcrtlcaof thc test sulxam_cemay. or are zuspec_l to, affect relatedmetabolicpm_ (c._. c,dcium,phosplme, luting uigtycaidce and eu_ng giuco_ specific honnm_ _). _y I mL ofblood will be collected into ]_)TA-coatcd tubes and _ ice or refiigerated until _ formLlym oflhe fDllowinghmna_lol_ on wet F.ayllm_yCtmomt (RBC) tIeumoctit faC't3 Henmoglobin(HGB) M_m _ Heeaoglobin(MCH) Cmlmmslsr Hmaogtobin CoucmaumionCMCelQ Mce_aCorp_ Vohmm 0VICV) LeuroWte Couat.Total (wBc'j Leukocyte Count, FlmelcCtount(PLAT) Mmm P_elet Vokmm (MPV) Cen ldomhology Two blood _naearslidm w/IIbe prepmedat the Tc_dng Facility for each sample for ofdifferemial leukocytecount. All munpics(on wet |cc) and tlides (mbient .... coalition) willbe_ toRed,oldLaboratorim atthe following address. Approximately 1.8 mL of blood will be added to a tube containin8 0.2 mL of mdimn citrate (0.129M_ The contems will be mixed m_dmintainedon wet iceumLlthe uxbmm_ umu_t Exygen Research Page 69 of 153 418-028:PAGE G-70 Exygen Study No.: 023-072 lhlloo1411F021 PageIS "" (within30 minutes of the coUcctiontime). Thex=sultingpla._a will be _ 2.0 mL polypropylme tubes labeled with studynumber, Sponsor's studynumber,ratnumber,dosage level, day of study, collection imzrvat, dateof coUection, species, generationand storage. All sanapleswill be fzozen on dry ice andmaintainedfrozen _-71PC) until shipmmt on dryice by overnight courierfor memma-tmaemofpm_ enembopias_time(APTa3. lime (PT) and activatedlmrfial Approximately2 mr. of blood will bc collected into senama_a'ator tubes and cmtrifused. The resultingsent semplcs will be immediately frozen on dry ice and maintainedfrozm _-70eC) untilshipment formudysiJof the following pmametaz: TotaPl roteinfrP) TriglycaidefrsR_ Albumi(nA) CrestininKeimts(cCK3 AlaninAcmimUm_eras(Ae t,T) Aspm_cAminotramfcrlaAscST) Globulin (G) Alktlinc Plmsphatuc (ALK) Albumin/GlobulinRatio (A/G) GX_se(OLU) Caloium(CA) _ (PHOS) Cholesterol(CHOL) Sodium (NA) Toad BilinsbinO'BILI) " Urea_tmgenfeUr_ -- Creatha_(CRF.AT) Potas_um (K') C_knid(eCL) Samplews illbeshipped(ondryice)toRcdfieldLaboratoraiettslafollowinagddress. _m.ltntsg_: Samplcs will be shipped to arriveon MondaythroughFriday accordingto the conditions dcscnT_cdabove to: PrincipalInvemisat_. Ms. Phyllis Powcll Redtield Labomories A Division of CRL-DDS 100 East Boone Street P.O. Box 308 Rcdfickl, Axkmnsas72132 Telephone: (501) 397-2540 Telefax: (501) 397-2002 The recipientwill be notified in advanceof sanzplcahipmcnt. UmNALYmS: Ufinalym will not be conducted unlcss indicatedbased on expected orobserveAtoxiciw of _ test mlbstmze. Exygen Research Page 70 of 153 418-028:PAGE G-71 Exygen Study No.: 023-072 "" M_THOD OF SACRIFICE: Pxotocol 418-028 Pa$c16 Fo generationramwill be sacrificedby cmbou diox/de asphyxiation. GROSS NECROPSY AND HISTOPATHOLOGY- Fo.,.G_.. _TION l_kTS: Scheduled SaCflfi_e- Tox/coktmetic S_dy: Schedu/edsacrifice of maleratswill be conductedon day 42 of study. Scheduled sacrificeof female ra_ wi_ be co_ on day 21 of lxesum_ gestations. Bloodmunpleswill be collected fromthe ratsas previouslydescribed. After u_'ific, thc liver of each ratwill be _tclsed _ the liver weight will be recorded. _ medianliver lobe will be frozen andstored (_<-200Cu) ntil _ment for malyl_is.Fetal samples willbcollectacsxi prfviousl_y Cmcasscs will be discarded withoutfurtherevaluation. Smnpleswill bc shipped on dry ice to l..isaClemen atthe previously cited _lxess. Scheduled Smnriflee- M_Im Study: Scheduled uca'ifice of malerats will be conductedon the day following th1_ _ admialsWatia_M_.efa minimumof 42 dayl of dosage. ScheduledL_'Lfice of female ratswill be conduatedcmday22 of laztatlon. Gramnecropsyof all maleand fmutle ratswill includem initial physical examination of cxtcn_ surfaccsroodall orifices, as wcU as the crani_ thoracicand abdomJn_c.avitics_ _ eontenm Specialsuentiomwill be paid to theorgans of theregoduetive systmn. _ numberof iuapl,l_ntafiotla_tes andcorporal',a._N,wt illbe _ Male and female ratswill be examined forgmu lesiom. Greta IcsionawiU be retainedin nmmal buffm'ed10%fonmdin andexm_nmxlhistologically. Tissue Wmm_ugandl_topa_logy wiUbe pcneormcdunderthesupcavisionof orby a Bomd-CmlifleVdetm-inm7 Pathologist. The ovaries andthe uteruswith cervix of each fmnaleratwill be weigbed, end ova'iu, u_ vaginaand amm_marygtandwill be reta/nedin neutralbufferedI_. formalln. Uteri of appare_ly_gnant ratswill be examined afterbeingpressed betwcm glass platesto confirm the absence of implantationsites, madretainedin nm._'ablufferei0d% f_ Exygen Research Page 71 of 153 418-028:PAGE G-72 Exygen Study No.: 023-072 Protoco4l 1_028 l_ie 17 _" Sperm_ns of MaleRats: To assess the potentialtoxicity of thetest micle on the male reprod._ve system, the endpo/nts listedbelow will be evahmtedflorathe firs_10 male ratsin esch dosage group. OrsanWeights: The fonowing organswill be individually weighed: right testis, Icft tmtis, lcft q_ididymis (whole and cauda),right pididymls, seminal vesicles (with and without fluid) anti prostate. Spm'mEvalualions: Spermem_-ntmtima madmotility will be evaluat=d_ compS=rassistedspermanaly_ (CASA). Motilitywill be evahuttedby the HamiltonThome IVOSby collection of z sample _mn the left vu d=_=ns. A homogmate will be. preparedfromthe Id _ _idymis forevaluation bythc Hamilton Thomc IVOS to deten=inee_mn cancmh-ation(sperm pergramof _ weight). Theremainmg"pordan of the left eauda_pididymlswill bc used to mana_ly evaluate sperm morphology. Spermmorphology=valmai_ _II i_l_Ic the followinr. I) det_inati_ of the pe_taSe ofnomud spe_ in a sample of at least 200, and 2) qualita_ve evaluationof almormal sperm, includ/ngsuch categoriesas abnormalhead, abnormaltail, and abnormalhead and tail See ATTACHMENT4 foraddflionaltiuu_ to be weighed andretainedfrom the tenratsp_"sex pergroupamgned to histologicalsample collection end evalt,_tfion. All othcrl_sucs will be disce.,d_ _lheduled 8n_lee ef Female Ra_ _xt De Not Deliver Litters: Ratzthatdo not deliver a litterwill be utcrificedon day 25 of presumedgcstatiou. necropsy,examinationamdtissue r_nfion will be conducted as dcscn'bedinviously _orrats_t scheduledsacrifice. with No SarvtviaE l_ns: Dems withno sin-rivingpupswill bc sacrificed_ Ihe last pup in found dead, raising or reed _'baliz_ _ rosy, _nmin_i_n _d _e _tenfion will be cm_iuctai as dcscribai abovc for ratsatscheduled sacrifice. Exygen Research Page 72 of 153 418-028:PAGE G-73 Exygen Study No.: 023-072 l_xo1418-o28 "_ Ibm Fond Dead or Man'bun" Kats th_ dior ml sacrificed becmxseof mmibtmdcondition, abm'tionor _ dc_ _ be examined for thc causc of deathor monl_nd condition on thc day thc obsavation is madc. TI_ ramwill 1_ cxamincd for grins lcsions. Tnstcs and cpiclidymidmof malerals will be exciscd andpaircd organ wcishts will bc rccordcd. Thcpididymidcswill bc _ m nmm-al buffered 10%formalin. The testes will bc fixcd in Bouin's solution for48 to 96 hours andthen retainedin neutralbuffered 10%fonnali_ Prcip_ncy status and utea-inecontents of fcmalcrats will bc n:cmdcd. Abortcd fctuscs and/ordclivm_ pups will be examinedto theextent posm'blc. Uteriof apparcntly nonprciPmmramwill be examined xflcr bcing prcssedbetwemaglue plates to confirm thcabscncn of implantationsitcs. Ovarics and uteai will be rctainal in ncutnd buffmcd 10%fommlin. TE,q_S. A/qALYSES AND MEASUREMENTS - F1 GENERATION: PrcwcaningPcriod: Litlcrs will be observed for dead pups at least twice daily. Thc pups in eachlittez will be counted once daily. _'ltuicsl (]_ser_atiQeuds/or C,mer_ Anuesraa_: _ Period: Once daily. Pupswill be observed ifthcy arc warmand clcan, for eviden_ of a nest and if Impsare grouped Uol_acz and nursingor bavo m/Ikin stomach. Pa, ch pup wUlbe examined for Snnendshape of the head, mink, lin_s, tail and_ of mus. Clinicoabl6crvatimoarym bereconiemdm-efr_qlumftlhyan_itcadbovoi,f deemed appmpdate by the Study Director and/orthe Study Monitor. Boar Welehts: Psi-mining Paiod: Da_s t Coinh),S, t5 and22 pos'cpm'tm. Saca'ifice: Tmminalweight. lVc-.eCdommmuttoa Valmm (rccanled and tabulated): Pn.vcaning Paiod: Not recorded. METHOD OF SACRIFICE - FI I_NEi_TIO N PUPS: FI gcnerafipmulpswillIx:ucr_ccdbycarbodnioxide asphyxi_ion. Exygen Research Page 73 of 153 418-028:PAGE G-74 Exygen Study No.: 023-072 Proll_oi 41b.02ll Pul_19 NECROPSY- M _gNERATION: Cae_ Imiemwillberetainiendneutrabluffete1d0% f_ forpoesiblf_utur=evaluatie_. uz_u s;x=i_r_y cit=dbe_w,anoth_tiss_ wiUbcdis_d_ Pare Found Dead on Dsv I Postoa_m: Pul_tlnndiebeforce0mmimni_ofthelittfmorpupviabiliwtiyllI_mndual_lfor_ _ I birth. The lunp will be remeved and immer_ in wat_. Pulnwithlungs that_nkwillbe _ed as stillborn;pups with lungsthatfloat will be identified as livcbom, and to _ _ shortlyafterbirth. Pup6with groesIm f_e_re eval_on. willbepreserviendBouin_mlutioa forpestle hm FoumdDead or Moribund omDram7,to 4 Ptq_ found deador sacrificedbecmm:of monlmnditywill be eyamfinedfor grog lesions end for the cause of deathor the mon'bundconditiml. Pupswith goes lesiom wall be preserved ha Bonin's soltdion forl_-m_le fu_e evaluation. Scl_laled _a_rif_: On day 22 pompaxtum,pt_ will be will be mcrificed and examined forgron lesi_ _ lesionswillbu_edinnealral _ lO%f_ _willincludeai_lle c_t- section of the head atthe level of the fiental-parietalsutureand examinallmaof thecross- seclioned brainfor _m'mt h3nkocel_ady. Blood mmplm will be collected fi_m each selected pup(5 per _ez p=- litter fi_madae 10 fmudes pergroup selected farFOB and mot_"tet/vity msmsmmt, blood samplecollection for I_C, and histological evaluatiem) from the vma earn. The blood will be placed into tubes,pooled per litre, allowed to clot and spunin a cazlzifiq_ then:_lling _t'tma will be I_nsfen'ed intolabeled polypmpylene tubes. AJIsampteswill be immediatelyfrozen on dry ice and maintainedAozen (_. 70"C) until shipnumtforanal_. The liver f_omeach _lected Impvdll be collected excised andtheorganweight_e_rdcd.The medianlobewilble frozmandstore_d-20%-'u)ntil shipmem for possible analysh. Px'ozenrun,lea will be _ to LisaClemen atthe l_-ViOUSly cited address, The n:mainias portionofeach liverwill be retainedinnetaral baffered 10% fm'malinfor lmsm'blehistological evahmtio_. Theliven will be proceasedand evahmted histologically u deacnbedfor IheFo geae_atie_ratsin HiatologicalEvalualiimna ATrA_ 4. ExygenResearch Page74 of 153 418-028: PAGE G-75 Exygea Study No.: 023-072 A PBOI_SF_.D STATISTICAL TF_SI_'tt: The following sdaemmic rqm_n_ mai_i_ nnaly_ of the dam Pnxlleol 418-0_ I I I I ! I I I ! I m. _ Trot_ H_og_ oftlmSlnon_aDlhlt_bu_n s. StafisCi_ly si_ficant pmbabilifies &e repmted as _qther p __ O.OS or p _ O,Ol. b. Propavtion data &c not included in this catcgory. c. Tcst for homosc_fity of variance. Exygen Research Page 75 of 153 418-028:PAGE G-76 Exygcn Study No.: 023-072 Pt"o_)_1411-0211 PaI_21 Teat ite_m in the FOB usin8 intm'valredes, such M the gr_gth trotsand the landing fuot splay test, as well as body weight data madfeed consumptionwincs will be tmalyzcdas describedtmdcrtim Pm'mn_c hcading of tim schtmutic. Bartlett'sTcst of Homogcneity of Variancest_ will be used to estimate the probabilitythattim grou_ had d;ffe_mt variances. A nonsignificantresult 07>0.001)will indicatethat anassumptionofhomogemity of va0r0ia)nccis not _, and the datawill be compaml using the Analysis ofVariance Test . Ifthat test is significant (p__.05), the groups_sed to the test zrticledsubstancwill bc comparedwith tim mntmlgroup using Dmmetfs T_ttmy. If Bartl.ett_Test is significant (p_.00I), the Ane.lysis of Vm,iuncc Test is not sq_zopfiam, andthe datawill be analyzedu descn]_edunder the Noeparametlicheading of theschematicW. hen75% or fewer of the scott, in all thc groupstu_ tied, the Kmskal-Wallis Tts_t2)wilble wed to tmalyzcthe data. and in the event of a significant result(p_O.Ob')D, rum'sTe_ TMwin be treedto umnlmrethe groups exlmted to the test m-ticl/subdan_ with the4t_.I.g.roup.Whm morethan75% ofthescoreisnany groupare tied, Fishers Exact Testtt } willbe used to comparethe proportionof tics in the groups. Data fromthe motoractivitytest,withrepeatedmeamrfe__ ts within a se.ion, will beanalyzed using anAnalysis of Variance with RepeatedMt=sut_ _3,as describedunderthat headingin the _cbematic. A tignificant effect (p_<0.O$i)n that tast can appearas effect of Co--on (a diffm_encbeetwem groups in the totalacromall mcasuremcatsin a session) orm an intmm:lion 1_-,tw_CnonccnWafionand Block (a diffm'm_between Ipoupsat _.cific meamretn_ --- pmlods). If the Cmmentrationeffmt is dgnifiramt,the totals fer tbe camtml grmxpat_ithe groups givctt the test articletsubstauncwill be comparedudng Dummtfs Test. If the Conccntrafionx Block _ is significant, an Analyuisof Vmianee Test will be uasdm _unt= the data st each _ period, and a sisnificant result (FB).05w)illbefollowed by a compari_m of the_oupsusingDmmetfs Test. Test items in the FOB having gradedorcount scores will be tmalyzedusing the procedures d_a'ibed readertheNotmant_etri; _ng of the schematic. Clinical observation iunidet_c data, as well as the descriptivem_dqmmtal datafixmathe FOB, will bc _ as ea_ntinget_ytables usingthc Variance Test forHomogetmityof the Binomial Dbtribufion (m. Alternateor additional statsticalevaluatiousmay be _ed if deemed necessaz3,or Exygen Research Page 76 of 153 418-028:PAGE G-77 Exygen Study No.: 023-072 Ptmac_418-0211 Page22 D_'I'A AcomsmoN, V_mmCA_ON AND_rORAGE: Data generated duringthe _ourseof this studywill be recordedeitherby handor using the Argus Automated Data Collection and Ma_=gement System, the Vivaritm Temperature and Relative Humidity Monitoring System, the Coulbourn Instruments Passive _ Motor Activity System, the Coulbourn Inatruraents Auditory Startle b_stem, the Coulbourn lnatrummta Spatial DelayedAlan,nation System, and/or the lumive avoidancemttwate. All datawill be tabular, stmmm-ized and/or statistically analyzedusing theArgua Automated Data Col/ec.aon and Managemem $yaten, the V'_/'mperature and Re./at_ Hu_/d/ty Mort/tot/rig$ysun,_ Mfcrazofl Excel Lnarot f MicrosoftOf_e 97 (version SR-2)] end/or Tht S.4S _t_ (ve_ion6.]2). Records will be n_ewed by the StudyDireb'torand/c_appmpri_ _ penonne/within 21 days aftergenera_n. AUoriginal r_ords willbe storedin the fchives oftbe Testing Facility. All originaldala will be boundand indexed. A copy of all rawdatawill be supplied to the Sponsor upon request Preservedlissuss will be storedat tbe Testing Facility atno charge for onc year aftermailing of the dr_ final_port. afl_ which time the Spousorw/_ be contact_ m determinethe disposilion of these materials. _-- KEY PERSONNEL: Exec_ve DL-ectorof Resea_h: MildredS. Christian,Ph.D., Fellow, ATS Din:ctorofR=seamlu AlanM. Hobennan, Ph.D., DABT Asso_ae Directorof Rusem_ andStudy Direcloc. Raymond G. York,Ph.D., DABT Direaor of Opemtious and Compliance: BarbaraI. Patterson, B.A. Dir_tor of_ Operation_ JohnF, Batne_ B,S: D/rectorof Study _ Valefie A. S'ba_, M.S. Managerof Animal Operations: DenaC. Lebo,V.M.D. Chairperson,Institutional AnimalCareandUse Conm_tt_: Douglas B. Izam, Ph.D. Consultant,Veterimu-yPatholo_: W. Ray Brown, D.V.M., Ph.D., ACVP Exygen Research Page 77 of 153 418-028:PAGE G-78 Exygen Study No.: 023-072 _" RECORDS TO BE MAINTAINED: Protocol mad Test Substa_.. Vch/l rod/or Reagcot Rzceipt, _on AnimalA_/o_ R_ion_on _cs. _nB Hi_ry. Trea_ (if prescn%ed by Staft V et_in_/nn). Gmer_I Commits. ClinicaOlb_rvsfiom rod/or_ BodyW_tL FeedC, onsmn_onVslucs. NaturalDeliveryObservations, FOB m_dMotac Act/viW Observ_ons. Blood Sm0zploCoLlecdon,_ and Sh/pmcoL Gmu Necz'epOsyb_z'v_ons. orsm wei_t_. Photo_-_hsf_ req_). StudyM_in_m_.,e(roomandenvlsumnentartecords). Feed and Warn"Analys_ p,cl_us ,rid/orSl_pmc_list,. and Use_ Protoc4o1_8-028 Tha Study Director will provide pmiodi updat_ of studyprogressto the Spmssur. Draft summary rabies ofunaudited compu/er-recordcd data may accompany/hose updates. Statistical mmdyseswill not be perfomlcd on these/ntez'Jmdata. A cmnlnlm_vc drs/t fl_l n_=mt w_I!bc pr_lm_d on complcfion of tl_ _ _ _1 _ fiml/zed follow/rig consultation with the Sponsor. The report will include the followin_, Sumnuuy mt Conclusica. E=paimco_ Design and Method. Evaluation of Trot Results. Ap_: Fisurcs, Smnmm-y and Individual Tsbl_ Protocol and Associated Amemlmmts md Deviation, Statemcot, Reports of Supportin S Data 0f appmlxiatc) Summ_ thc Abovc Data, Study Director's GLP Compliance and QAU Statcracot. The Spomor will rccc/vc mac copy of the draft n:port madtwo copies of the final report. Data will b frond- and/or computcr-rcconicd. Records will be rev/ewed by the Study Dir=ctor msd/or appropdatc _ pea'scmnel wilbin 21 dnys after scncn_on. All original records will be storcd in the archives of the Testing Facility. All original dam will b bound and A copy of all raw da_ will be suppEcd to the Sponsor upon request. Ps_scrvcd t/ssucs will be stored at the Testing Facility at no chin,go for one year O.er_ of the draft final ,-- _cport, after which time tt_ Sponsor will b= com_tcd to detaminc the dispositim of _ Exygen Research Page 78 of 153 418-028 :PAGE G-79 Exygen Study No.: 023-072 l_'olacal41g-0"2g _24 INSTITUTIONAL ANIMAL CARE AND USE CO.MMn_'EE STATEMENT: The proceduresdeecn'bedin this protocolhave beea reviewed by the Testing Facility's ImtRutic_d Animal Careand Use Committec. All pmeedur_ du_fi_l in thh protocol th_ involve study animalswill be condtu:tedin a rammerto avoid or minimize discomfort, d/_._s or pain to the animah. The Sponsor's signature below docanumtstim fact that_n cvneeming the necessity for conductingthisstudy and the fact that this is not an mmec_muily duplicativeetndy may be obtainedfi'omthe Sponsor. No altmmfive(/n _tro) procedures weze aveilable formeeting the statepdurposeosfthestudy. mz_zmz_c_: I. Chr_ I_LSm.xlVoytekP_F,(1982)I.nF'rcoRepr_andMu_g_U'yTeat_. Envimmnen_ Pmtec_onAge_y,WmhingtonD,.C_Nation_Technicallafen_tion Scxvice, U.S. DepmmumtofCommm_ Slningfiekl, VA 22161. 2. Christiart,M.S. (1984). Reproductivetoxicity a-d tera_logy cvaluelions of naltrexone _81 of Naltrexone Symposium, New York_y of S_ie*_s, November7, 1983), L Cliv. I_hieL 45(9):7-10. A 3. Lmag,P.L.(1988). Embryo and Fetal DevelopmeatoJ To_e|ty ('Fermolo_) ControlData /_ t/_ C_ar/es P/per O/:_ P,a_ CharlesPdver Labomtodes, Inc., Wilmington, IdA 01887-0630. (Databeae provided by Arg_Resamzeh LaboraIories,Inc.) 4. Imtitute of La_ AnimalRemurces (1996). Galde for the Care and Use of LaboramryA,.:/_/_. Nafio.a/AutdemyPre_ Washington, D.C. 5. I_, G,C. (1989). Developmont of Ti=rI nemobehav-ioraltesting capabilifimfor incorporationinto pivotalrede_ safety auessm_t studies. J. Amer. Col. ToxicoL8:53- 70, 6. Irwin, S.(1968). _vcobse=vationalames_maont: IL Asystemioquantitative pmccdureforasseuin 8 the behaviorsland physiolosic state of the mouse. Psye..lx_harmacolosia(B_lin) 13:222-257. 7. Mose_,V.C. (1989). Scre_in 8 approachesto neurotoxii_. A functionalobservational battery. J. Amer. Col. ToxieoL8:85-94. 8. ODonogh_JJ. (I989).S,a_ahagforneemmxiciutysinganmmlogicallbyued examinationand neumlm_logy. J. Amer. CoL Toxicol. 8:97-116. 9. Sokal, R._ ondRohl_P./.(1969). Bsrtlett'stest 0fhomogme_tyofvm'hma=a. B_omefry, W.H. FreemanandCo., SanFrancisco,pp. 370-371. Exygen Research Page 79 of 153 418-028:PAGE G-80 Exygen Study No.: 023-072 I%omm4ll_28 I0. Smxi_or, G.W. md Codm_ W.G. (1967_ An_y_ ofVw. _a_ 6th F.di_ Iowa St_ Umv_ _ Am_ pp. 258-275. M_od3, 11. Dmm_C.W.(1955). A muRiplmmp_um progedm_ for _ng s_ral _,_m_mts with s cona-ok J. Amer. Star. Assoc..50:1096-112l. 12. SoksIR.R.m_dRohlfF,J.(1969)K.ruskad-WalTlmi_s. B_,,etv)W,/,-I.Fr1_manm_d Co.,SenFrancisco, pp.3819399. 13. Dmm, OJ.(1964). Multiplecompmisomtmingrsaunmks.Tc_Imom_xi_6(3):241-25Z 14. Sk_l. S. (19S6). Nonlmoame_c Smti_c_ fov the _e.lm_oral Sciences, MvCm, w-I4ill, New Yc_k, pp. 96-104. 15; _lustitute, lm:.(1988). I_pea_rc_um'_aualysillofvazim_e. :?J_STATTuUseWa C_ide, Release 6.03 Edition, C,aryN,C, pp.602-609. 16. S_xlc_r, G.W. a_l _ W.G. (1967). V_m_ V--stforhomogcm_L7of th binamial _tx:tbufion. _m_s_cal MKhodz, 6th Ec_fion,Iowa StateUniven_ Ames, pp. 240-241. Exygen Research Page 80 of 153 418-028:PAGE G-81 . Exygen Study No.: 023-072 PROTOCOL APPROVAL: FOR THE TESTI_G FACILITY Alan M. Hobcnn_ Ph.D., DABT Dinmt_ of Rtmmrch l_-'ladL_418028 Da_ -- _ Memb_,lasli_-fioualAaia_Cm_d Um Commim_ FOR THE SPONSOR lo/m Buamhaff_Ph.D., DABT, CIH Date Sma__mr a_l Exygen Research Page 81 of 153 418-028:PAGE G-82 Exygen Study No.: 023-072 ATTACIDIENT I SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE Exygen Research Page 82 of 153 418-028:PAGE G-83 Exygen Study No.: 023-072 A'I"rA.CHM_|I'r _b'TUDYSCHEMATIC l_toco1418-0_ PagnIof3 COMBINED R.EPF_.,ADTOSE A._D I_PRODUCTIVF.,_EVELOPMEI'_A.L TOXICITY SCREEN' L ForKldiflom_fl see"T_IsA,mfl_ andMe,a_ments" scctloonfthcpn_,ol. b. FOB andmotoractivintvyah_onz _ ontm mMm ixag-ruup. c. Male rat* mcriflcedafla- completion of at l_st 42 days of dora, g_, n_Zol_y and ofnmk roprodu_ve organs. H_mtology, clinical biodmnistry and histological m_ples c,onoctodmmatm realratsper group. d. Taa fcmalo r_ 1_ Szvup_ignad m FOB _'vahu_a and m_ _vi W e_aluafi_ e. Tm _msl r_ per l_p m_lthnir littcwssacrtfic_ion day 22 _ necrop_ _ nn_a_ionoffcm_st__ argem. Hematology, c"lmicabl iochemistry and histological samples collc_-t_l. _ md diatoniC. female rats sacrific_l onday 22 postp_Uma Exygen Research Page 83 of 153 418-028:PAGE G-84 Exygen Study No.: 023-072 ATTA_ I Prote4c1e8d-028 Plg2eor'3 SCHEDULE" 26 MAR 02 01 APR 02 01 APR 02 - 12 JUN 02 14 APR 02 PM - 21 APR 02 AM 21APR0_ PM- 28 APR 02 AM 14 APR 02 15 APR.02 28 APR 02 06 MAY 02 19 MAY 02 06 MAY 02 - 0"/MAY 02 AnimalReceipt -AcclimatiloanegiDi. Startof Dotage Period- Male Rats (14 dad before cohabitationandthrmtgha 14*day_aabitati_a perioduntil the day before samSfioea_er at least 42 daysofdosase ). Dosa_ Period- Female Rats(14 days before mhabitalion timmgh Day 22 of la_ation). CoimbilationPeriod(Maximtmaof 14 days). Male I (7 days) Male 2 (7 dayt) Day 14 Toxico, kine_ Sample Coileclion FixatPosm'bleDay 0 of Presumed Last Posm'bleDay 0 ofPtemm_ Gettatio_ First Posm'bleDay 21 ofPreeumed Gest_oa ToxionkinetiSan_k Collecdon end SacrlficeToximkin_c Study Female Ra_ Last Poem'hieDay 21 of l_mmmaedGestation ToxlcokincticScruple Collection madSacrifice Toxicokinetic Study Female Rats FOB andMotar A_ivity Evaluation - 10 Male Rata pa Group a. Thc'atmt,date of the m3m'hy lhe day the Study _ iisns l_e proteeoL b. _ lid,, _.bedadetl,_ din]ofbixthisdedipaaledayI poctpax'am(daayI of la_nion)aa__! _oKq_m tip ot'tlaFe1ipmm'_ormo u4 daysofthebctation -'_ period will be detmmfiztedand cited aco_rdingly,emdaacribedabove the proto_l scctlon, _bedn8 S_em." Exygen Research Page 84 of 153 418-028:PAGE G-85 Exygen Study No.: 023-072 ATTACI]_fENTI "-" 06 MAY 02 23 MAY 02 10 MAY 02 23 MAY 02 12 MAY 02 t3 MAY 02 23 MAY 02 - 09 JUN 02 27 MAY 02 - 13/LrN 02 24SF.P02 Pmtoco/418-028 Pl_ 3 or'3 FirstPo_'ole Deliver7 (Day 21 ofprcsumed seaat_b). Last Possible Delivery (Day 25 of presumed _on). Fi_ Pore'hieDay 25 of PresumedGestation Last Possible Day 25 of Primmed Gestation Female Sacrifice, Day 42 Toxicok/net/GSample Collection and SchcdulcdSacrifice- Toxicokinetic Study Male Rats Sehed_ed Ssc_ce - MainStudy Male RatJ (Earliestpmsible date). Hematolosy, Clinical Bioche_stry andHistological Sample Collection of SelectedMale Rm. FOB andMotorActivity Evaluation - I0 Female Day 22 P_ - Sacrifice Female Rats and 1-k_tok,gyC, tiai_ Chemi_y-,,,4 H/_los/cal SampleCollect/on of Selected Female P.m mdPup.. DraftFinalRepro Exygen Research Page 85 of 153 418-028:PAGE G-86 . Exygen Study No.: 023-072 ATrAC]8[MEI_r 2 MATERIAL SAFETY DATA SHEET Exygen Research Page 86 of 153 418-028:PAGE G-87 Exygen Study No.: 023-072 .,. ,- i I'LATERtI_LSAFETY :_q OATA SHEET el4 Center (Exper_mentLl) St, Paul, _nmota $5144-I000 1-800-364-3577 Or |051) 737-6_1 (24 hours) Copyright, 19eS*, HtLnneaot_ f,tLn.lng and I,tmsul"iL_'t;ur./.nOGompln, y. A11 r_Ohti reserved. Copy_ and/or d0t,'n.'Lold,l.ng Of tJlll n?ormmton for The pwrpe|e of properly Utd.liz_ 3H pro_l 1= LUm4ed prov_d thst: 1 ) the _ffom'tJ.olr) is _-ep:Lod:in 'fuZ1 Ntth _O chXngan _leae prior _reemo_t _ obtmed from SH, and 2) neither the copy nor the oriolna..?. :be r@llo.T_dor ;tllll, tZlme dlstrJJ_ted wAth the dJ_ewt_m of eern_ng a profit thereon. 01VIe[ON: _ 8PEOZALTY MATSR_LS Ft_TERIAL: L-imSl OEVELO_ENTAL F_OOUCT ZSmNUD: December 07, 1D_P SUPEASEOEB"_y 17, 1009 DO_UI4ENT: 04.8470 -2 ......... . ....................................... 1. INGREDIENT ..... . .............. ..... . ................ . .... ......... ......... .... .A.8. NO. . ......... . ..... PERCENT .o ...... . .... . ..... POTPaSZUPt PE_UO_ SULFATE ..... RESZaI_ ORGANIC FLUOROCHB_L8 ........ _71-gg-e H.txtur_ 100.0 Unlmm_ Ths mmtord_1 Is not listed on the TSCA lnvonto_/ and thou/d for reHxreh and devalopmen_ purposes only under the d_rec_ supervision of | te_hnlcxlly qualLfled tnd_vJ_luLt, be uxed ............................ 2. PHYSZCAL DATA ......................... ..o. ........ . .......... .. ....... o.. ............ . ...................... . ................. ..o . ......... 6OZLZN_ POINT: ................. VAPO_ PREUU_: ................ VAPOR OENSZTY: ................. EVAPO_AT%0_ RATE: .............. GOUJSZLZT_ ]IX _TER: ........... SPECIFIC _qAVTrY: .............. VOLATZLE: .............. I_t: ............................ VZSCOSXTY: ..................... HELTZN_ POINT: ................. APPEARANCEANO O_OR: 0t'l'-,,._h_.te orys_slltne solid, NIA Nei_T/Olble NIA kOltg_ll s_Lgh_ .. 1,0 Na'ter,,l (eulk) Nmgl,lg:Lble NIA NlO N[D sl_trp odor. ............... _revlatlmls: o..oo ........................................... NID - Not Oetero_ned NIA - Not Appllcabl$ ..... .oo ...... . CA - Approximately Exygen Research Page 87 of 153 418-028:PAGE G--88 Exygen Study No.: 023-072 MaDe: L-9061 _cmmbsr 07, DEVELOFflENTAL PRC(X_'T 19_ PAGE 2 3. FIRE AND EXPLOSXON HAZARD DATA o. ................................. o............................... I_ZNT: ................... F_LE LZK_'P3 - LEL: ........ LZHXTS - UF.L: ........ AUTOTEIITTIWi TEtqPERATURE:...... :, _12 F htaf2ssh Setafluh N/A elslid Cup NI^ N/D o ...... oo. _mr. earns, dtoxX_. Dry _L_Z. Fob- SPECIAL F_FIE FZGHTZIM PROCE(XJlU_: Mear'fu 11 protective Ol_hJ_ll, AqcXvdAng ho2Jt, seX1'-c_a_ned, posture pressure or pressure demmd brat_AnO apNrstu, banker and plml_a, hand around Irml, _ and leg, faoe uk, aml pratct/ve ooverJ_: for exixmd area _' the head. co_ UNUSUNLFXRE _ EXPLOSZON _: _ Hazardu Decompoe1:lon l_tJan for produou ot' CeJsutlon. ........ ..... ........ . ............................. 4. RF-J_TI"VTI_f DATA ......................... o......................... .o..o... ............. _oo.o . .... **. ...... . ........... 81"NIZLTrY: 8table XN{_i_PATZBZLTIrY - MATERZALS/CONDZTZ_i8 TO AVOXD: None knmm. POLYI_R_.A'rzoN: HazJu_k_ poly_er_.zs2J.en i_Lll not cur. _ZTXON PRWXJCTO: Ray produc fluoroGarbon lever 3SO C). OUe 2f Xl_ld tO very h_h tetpertur o....... ............. . ..... . ...... 6. EHVXWOHI4F.flTNX. NFORP_TtON ..... ... ............................. . ...... ... ................................. . ............................. . ......... 8PXLL RSFONSE: Obsrw precaution fPoe other e_t_r_ns. Collect pLtled aatrla/. Us wet mileplno _oapound Or _atr to av_Ad dustAng. Clean up IPlll_lt_. PIIC:O _ :_oad i=olTtLnr. RE(;O_NI_D OTSPIMAL: Z_o:LnPl:e _J1 n :i_O_ll_r_lll Or _Omll_tl fC_.lty _fl _hff presence Of coabusttble aatar_aLl.. C_butt_ llrmtu_ts NLlX Include HF. OAspeal a_terllltAve: Olspo If 8it proE :Ln flP-4_l_y perllL'_tld to ll=_pt: chilli:=&1 lalrtl. ............................................... Abbr_vLatJ_ns: N/I) - Not OetraLn4d . .... o.... NIA - Not AppliCable o...oo. ............. CA . Approxllatly E.xygen Re,_areh Page 88 of 153 418-028:PAGE G-89 Exygen Study No.: 023-072 MOB: L-9051 DeclmDor 07, DEVELQ4q_KTAL PRODUCT 1999 PAGE -q ................................................... 5. ENYIAOm_rrAL .......... . ............ INFORHATZOU (continued) . ............................ 0. ..... . .................. . ........................ ENV_RQNHF.NTALDATA: NoT detemlned. REGULAll)RY ZNFORMAT/ON: Yollt:Lle Orga/llc Collpolmds: N/D. VOC Lces H20 & Except 8olwnts: N/D. Since regulatlofls before dksposal. EPA Hazardous). very, onsuZt applicable r_gu/al_oce or muthort_Lce U.8. L_'PAHazsrdous klmlts NuabeP - None {Not U.3. 0171Ut BWZllOlt'4ERrJtL ZNFQR_TT.0N: This produmt say cNta/n one or sore argan_ fluorecheaL_ls thl_ hays the potsntlLL .to resist degradation and pets:Let In the rl V:LrOfl/Ult EPGRA HAZARD GLASS: FIRE HAZARD: NO PRESSURE: NO RFJtCTXYTrY: No ACUTE: Yes CHIWNIC: No _ ..................................................... ..... ....o ..... ..o.. ....................................... o ................. EYE (_IITAGT: lmmd:Lately flush amdcel attentisn. eyes wtCh lsrge mounts uf _atsr. Get hmedatu 8KZN COt4TACT: Flush sklrl with lrge aed2ceZ at'_ent/an. Imounts o1"Niter. I1' /rrl_stl_ pere/st:s, get ZNIMLA'I'ZON: If sLgnsleynptoae c_our, r_ove person to fresh alr. f stgns/syaptces oontlnum, cell a qmyshd_m. ZF JtNJ.OHED: Zf led8110t40d, eL1_1 phys:LotJm tared:Lately. Only Induce roe/Sing st the Lnstruc_Lon Of i:hyslGisn, Never give a/tyth/ng by south to an unoonsc).ous _t rso_. ......... o... ..... o ............. o............... 7. PRECAUTIONARY ZNFOIb_IATZON .......... .... ............................................................... ...o. ..... o..o .............. EYE PROT_TT_" Avo/d lye _Itsct. AVOid eye c_ttilct _Lth vtporp lp('ay_, The folloNtn O should bs uorn mIOM or In r.ol_:bta_nf u _o prevent lye contact: I_ar rental goggiss. or SLIT. app_oprtats, ................................................... Abbrevisl;J.ons: NID - Not DetcrtLned NIA * _ . ..... ....o. ..... o....... Appl:Losbls CA * Appr_sisstsZy Exygen Research Page 89 of 153 418-028:PAGE G-90 Exygen Study No.: 023-072 Haas: L-NS1 OEVELOImENTAL PROOUCT Dacebar 07, 1909 .............................................................. 7. PRECAUTZONARY][IIFQPdflAT][Q_i (oontlnued) ................ ........ ..................... PAISE 4 o.o ............ . .............................. 8KZN PROTECTZOH: ^vo_ sk:Ln cent|or. Neap sppropr_te gloves wlllm Iw_ll_r_) th_ mater:Lal. A psr of gloves Imde frt_l the feline:Leg ntmrtal|a) ere reammtmded: butyl rubber, polycthylanelpolyv:U_yLUIime chlorJzle (8&_mmx). Use one or imr Of the fo].]_N:b_ tus u necessary to IS_VQn_ sk_ O_tL_C: para4)mll pro'=_ton he "d cavorino, cover_Llls. Protm_tve 9Mlen_s |el:her fJIn glovws) _ld N na4e of aLl:her Of _1_ ?011a4JJlg ImtorJdL1La'. polyethylenelpolyvtnylldlne chlor:Lde (8arlmox). RE_ VENTll_TZON: Usa N_ eppraprLate _ lom_L exhaust van'.J.tJt_b_ valll::Llated area. Provide ex_ut vmttLl_Lon. Prov_ie appr_.prLste at trimfar points. IJ_a in tm_L- auffL:/4nl_ vent//atJAn to uLnta_Ln uttaslmls helen rl_mmckld exposure 1/Lea. Zf axJumst ventklat:Lon 1: not edeq'tae use approprJ4to respiratory _toa. v_nt41stlo_ IKlewato to oof_ro_L vapor ooftoentPttone Prevldo I_lcyd re_ommnded exposure 1tad.re and/or _ont_lL eprsy or slat. LOC,l_ eXlIIU/at venttlatLon eel airborne. Is reommmdml _here t_n mlter18/ beco REaPZRATORYPROTECTZON: Avoid breal"Jtlng of LtrUorne Meat/el. 841act one of the f_ N]_OSHapproved reapreto_s based on a_;J_orne ;onc_ntrcTtan of contalll_ante a_d _ _rdllnml _Lth O_HA rl_gUlat:l_ns: flier-mink eupplLed e JJ- respJJ-al:or, full-face lupplJJId I_r respirator. P/_Ofl OF ACCXi_AL |HOP.STION: Do not eat, d_nk or imoka _hen using thai product. Nash exposed &reas thoroughly _th soap and rater, k_sh blade after handlkng an_ before eating. REC_4W_OED STORAGE: 8tore st reoe temperature. cXoaed _ne*l not In uae. KJap can_Ltnar dry. Xee? container FXRE A_D EXPLO_Z'DNAVQIDNII_: Not detemJ_edo EXI_E LI_Fr8 XNgRED][ENT VALUE UMtT TYPE AUTH 8K_N* FOTASSZU_I pERFLUOR(IqEXNIE 81JIJ_T_ .......................... 0.1 H_dH3 Tl4A 3H Y R_S][DUN.. 0RG_IZC F_CAL8 ..... II. t M6IH3 Tl_ 3/4 Y _'_ 8K:N NOTATION: L_stlKI SUbstanCeS ind/catocl _Lth 'Y* under SJ(ZN tiller _o ................. . ....... . ........... . ..... ..... ....... . .... .. .... . .... .. .... AbbrevUItLana: IllO - Not Daterteed Ilia - Not Applicable CA - ApprexLaal:aZy Exygen Research Page 90 of 153 418-028 :PAGE G-91 Exygen Study No.: 023-072 MS09:L-9051 DEVELOPHENTAL PI_0UGT 0ecember aT, ION PAGE S _Xf_)SU_ Ll_rrs luOn_inuea) TNGREDXEHT .......... . ........ ...o. .... .. ......... VALUE trait ..o_ ....... ...o.o..o _he potential contribution to the OVerlLll eXpOSU_I by the L_uludL_g alucaus acabrlms and eye, s2thir by mJ.r_r_u or, by dtrec_ course1; Nlth the substance, velllclss nan slur TYPE AUTH _TN" ..... . ............ _*u_aneous routs mrl partLcularly, ek:Ln sbsorp_Lon. SOURCE OF I[_OSURE LZHZT DATA: - SHe 314 Rs_oummded Exposure 9udel/nss ..... o.......... .. ........ 8. HEALTH HAZARD DATA . ..... . ............................ . ............... EYE CONTACT: No JJ_ro_t_on _8s found regarding elf sots fr_ Bye r._tact. 81ng].e exposure say cause: Hild Eye Zrrtul:Lon: slgnelsylptmls sNellLng, pale, end tearing. (.in :Lnolude _dnsss_ 8KZN CONTACT: No tm'omatton kms found regard/n 0 el_o=t_ frog skJa con_aot. Hay be ul:sorbed Ohrough 1_1s 8k/n and permLst /n the body for an extended time. 81nolo exposure may cause: Noderat_ Nl12nO, Skln XrrAtstton: slgnslsymptoms Achlng, and drynmu. csn lnclube redness, XNHN_ATZ0N: No 2nforwatlofl _as found rsOFrdtng s/_l'sot:s from /nhalatlon exposure. Hay be absorbed 1:lie, by _nhalst/oa and perllist In the body for in sxtendud 82nglt overexposure, above red.ended Outdel_us, nay cause: [rritst_an (upper rQsp_,_tory): s_gnslsyeptoaS can _nclude soreness of "the nose and _ttraat_ _oughLng end s_eszlng. _F l_OkLLOHED: ][ngesto_ s nat _ X_ksly roues of sxposur_ to thLs produc_. No :_lfo_s_lon was founcl regerdlng effecOs frms s_llm_Lng. An2_sl studies _nduol:mi on urgsn1_ fluoro_t_ml_sls v_ttctl ssy be pr_lsent 2n this product _md_mte sf4'sots /nc2udLn 9 12vtr dls1_Jrl0snoss, _e2Oht loss, loss of sppst_ts, lmtMrgy, end r,eu_o2og_cal, pancreatic, _-- sdrans2 and hms_olo91c effs0ts. Tlnsr_ arm no known humus hea2_h ........................ . .... . ...... ..... ...................... _...... .... ... Abbrev2atlons. N/D - Not OsOsrmlnmd NIA - Not /Oq=llcabls CA - Approx:L_sl:oly ExygenResearch Page91 of153 418-028:PAGE G-92 Exygen Study No.: 023-072 A qSOS: L-gOSI OEVELOPHENTALPROIDUCT Oeclllloer 117, 1999 .... . .......................... 8. HF.ALTH _ ....... ...... ...... DATA . ....... . ........... (oontd_uod) . ..... . .......... o......................... . ................................ PAGE e ........ errors l'rse mntL_J43atod exposure to these filM:L fZuoroohelloLll _hen used am Latedad and lnatr_c_ed. OTHER flEALTH HAZARD ZNFOPJ4ATXOH: ThLs product amy oo_aLn one or ao_ orlpm_J: fl_oroohem_oals tl_t have the potent JJ1 to be absorbed and _ _ the body ?or long pit:Lads of t_llle, etber 141 the parent 1souls or as NtaboLttes, and aay aooumuJate k_Ltl_ repasted exposures. There are no known human health effects ?luorochme_.als from et;b:llMted eX_OOIUrOto these orpinlo _on used Is l_tendtd and Lnstructed. The presence of orgmnJ_ fl_oreoh_4a_Ls la the blood ef the ge_ral popuJJtJ_ll iiId lUl_llpli_lt_tlll, _ Ill IIIPI_PI. MS _ det2q hick tO the 1970'l. 3H'I ip/chHe_olOOJJ_/ study o1' _to ann Markers LndLr.ltas no adverse et_nots, .... .... . ....... ..o .................... GECT1[ONr._4N413EOATES ..o...... ............................ PRECAUT)_IDi_RY I"NFO. 8ECTZON _ SZI_E Hay 17+ 1999 ZeSLE ......... ..... ....... . ..... o..................................... N:)brev:lLal:/_ns: N/O - Not Olterltmld N/,i. - NO1:_.IJ.r.ablw ..o... ...... CA - ApproxLlately ..o. ........ ... ..... .*................ ........ ...... ....o ....... . .... . ........ The _rlforalatJLoll :Us thlll HIiter'JlJi2 _ety Ihrta hilt {1_o!1) la I_llltJl_qld tO be errat as of the date tallaid. 314 _ 110NARRANTZES, _ OR ZI41q.][EQ, I'NCUJOZNG, BUT NOT LIHZTF.0 TQ, ANY ZllPL|ED 14ARRAt4TfOF MI_i_HANTABILITY OR _ FOR A PAKII_CULAlitPURPOSE 011OOUME OF PERFON4_lk?_ OR _ OF TRADE. User tl rOllpOlll:l_le for datorlL1Ln:lJ0I ideaher the 314 prodL-' _ ft for palrtJLoullr purpose and lu_tll/_J for ueer's method of use of" app_tLcatlon. _Lvaln the var]Lety of factors that ran al_ect the use and pplcatJ_n of a 314 product, some o1' Nhch Ire unquely H:iL'thJ.nthe uslr's knOl_edgl e_ld GOIt_l'9_, _ _S essentLal that the user v_l.ulte the 3+1 prodc4: _ detlPlllml 14heher t :Ls fit for pllrtLOlJZiir purpole and ilUttlll_e for user's llthod of use or Ip_l:Lcltlon. _4 provLdse _Jlformlt_on UI |lectren*o for as a lerwLe tO LtS C_e_oltrn. Due to the re_mte possJ_J.l_.ty that e_trm_la trsmsl'er my h_yl resulte_ in If'rare. oailslone or I,l.teret:L_ne in this J_formltlon, _q mikes nO _epresentlt_on as to tl completeness or c_r&cy. Zn _dd_to_, "" Lrn'orlmtdan obtained from database may not be ms _urrent as the lnt'erlllt:Lon 1/I the HaOtl avlLL_Jlbl_ d_LrllCtly from 3_. Exygen Research Page 92 of 153 418-028:PAGE G-93 Exygen Study No.: 023-072 ATTACHMENT 3 TEST SUBSTANCE PREPARATION PR_URE Exygen Research Page 93 of 153 418-028:PAGE G-94 Exygen Study No.: 023-072 A'I'rACHNENT3 _ 418-028 Verten: Page1of2 TEST SUBSTANCE PREPARATION PROCEDURE "TestSubstance: T-7706 Vehide: Aqueous0.5% CMC (mediumviscosity) A. Purpose: The purpose of t_is Woce_m is to wovide a methodfor the Weparation of dosage suspens_s of T-T/06 for oral (gav_e) aclministratl_ to rats on A_us Research Studynumber4t8-028. B. General Informatl_: 1. All suspensioncontainerswill be labeled and color-coded.Each labelwill specifythe protocolnumber,test substanceidentifical_n, Argus batch number, concenlndion,dosagelevel, preparationdate. exp_raUondate and storageconditions. 2. Suspensions wii be prepared: -- __ Daily _ Weekly _ For_ days of use _ Appro0dmatelyevery ten days _ By Sponsor 3. Suspensionswill be adndnistemdat a finsldosage volume of 10 mL/kg. 4. Safety:. Gloves, uniform/labcoat,goggJesor safety glassesw#h idde shkMdl Du_-mist_EPA.Rtemd Mask Half-Face Respiratorif not usedin a chemical fume hood FulkFace Reeplrator/P_,Jve Prem_m Hood Tyvek Suit or tyvek apronand sleeves 5. Dosage suspensionsadjust_i for % Acth_/Pur_y or Cowsct_ Factor. Yes X No (Calculatk:)nsbased on 100%) __ % _ __ % Purity __ Cormc6on Factor 6. Sampling mquimme_: Citedin protocol 7, Storage: Cited in protocol Exygen Research Page 94 of 153 418-028:PAGE G-95 Exygen Study No,: 023-072 AI"rAGHMEN3T Protlx_ 418-026 Vemion4:1J_02B|1M9 AR021 PageZ or2 TEST SUBSTANCE PREPARATION PROCEDURE NOTE: Priorto test substance preparationaccurately measure the required amountof the spprop_tm vehicle (R.O. deionizedwater should be used for calibrationpurposes) in a graduated cylinder,pourthe required amount of vehicle intoa beaker. Carefullymarkeach beaker at the meniscus. This mark wlJIbe used duringthe preparationto bringthe test substance slurryup m volume. C. Dosage Suspension Preparation: 1. Woigh the required amount of test substanceon a pieceof weigh paper or Intoartappmpristely sized mortar (see PREPARATION CALCULATIONS). 2. ff weigh paper is used, transfer the test substanceto an appropriately sized mortar, ff necessary, grindthe testsubstance intoa fine powder. Slowly add a smallamount of vehicle andgrind.Con_tue to add vehicle slowlyand grind 1hevehicle and the testsubstancetogstherto form a fine .-- =tufty. Transfer the vehicle/Im_ substance 81urryto a markedbeaker. 3. Rinse the mortar and pestle with additionalvehicleto removeany remainingtest substance. Transfer rinse to beaker. 4. Add additional vehicleto the beaker to bdngvolumeup to the rr_rk. Place on rrmgneffcstir plate and agitate priorto and duringsampling,aliquoffing ar_I/or adminlstm_n. 5. Repeat steps U) through(4) for each concenb-.tion. J't .,= Approvedby-,_J._/_ _ Data: Z_Poz.-- -(-j 0 Cla_catlon: _ No "-"" Yes [see at_ched clarificationform] inlt_l/Date : Ex, ygen Research Page 95 of 153 418-028:PAGE G-96 Exygen Study No.: 023-072 ATTACHMENT 4 TISSUES TO BE WEIGHED, RETAINED AND EXAMINED HISTOLOGICALLY i : ii i : Exygen Research Page 96 of 153 418-028:PAGE G-97 Exygen Study No.: 023-072 ATTACI_IENT4 Pmt_o4_1&-0_ PalPIu:f2 "" TISSUES TO WEIGHKD AI_ RETAINED FOR _ EXAMINATION FROM TEN RATS PER SEX PER GROUP The tea ratsp_ sex p_ groupassigned to fia_ional obsnrvationalbatteryand motor activity t_ will be assigned to _1o_, c]Lnicabl iocl_n_i_ andhislological evaluations. The following organs will be _cis_'xl, llimmed and individuallyweighed as soon as possible af_ excision to avoid fiver kklnoys adz_za_ spl_m brain heart thymus ovzries testes u_su (wi_e_vix) ri_t epidkt_ _utte lefi epididymiz(whole endcauda) seminalvesicles (with and without fluid) I.illlm..a.b_t.Kr_: The followi_ lissucs orrqm_nt_w smnplewsillbcretainiendn_alral b_ 10% formalin. brain(rqn_w resio_ includincget_orumce, rebellump,ons) and _ hm_ _ intestines ft_l_li_ lk-yCs patolm) vhthnmmdb_tffm_1d0%_-malin) lymph nodes (subnumdiblarand mediastinal) paipbaalnavc(sciaticortiblal) l_ons stomach spinaclord(cervicatlh, oracaicndlunar) liver kidncys sple_ hemt a'achca urinary bladder uterus bonemarrow (sl_"mma) ptostat_ ,nm/nol wtid_ (with _sulatin_ _Iml) mmm.rySl,.ad(r=,u_ratsonly) uterus valom * Teates will be fixed in Bouin's solution for48 to 96 hours before being retainedin ne_t_ buffered !0_ film, lalin. Add/fire.fly, theremainingpo_on of the lofetpididymis(corpus and caput),aswell as theright epididymis will be fixed in m_aal bufl_rnd IO'Aflmnslim Exygen Research Page 97 of 153 418-028:PAGE G-98 Exygen Study No.: 023-072 ATI"_ 4 Prelocol418-O28 Pal_2cof2 _" Itlstolot_IcalFauualnatlom: Histolo_i_l mzminatio_ ofrmi_d tissues, imludi_ rqnod_'ti_ organsw, ill be emadueted fortl_ mi_a_l ma ratsl_x from theeo_anoiandhigh dos_, Stoups and flora _c F1 geaeralion pups Oivm's)fi'om the control andhigh dossgc groups, lXlcsions alm'butedto the test subs_ac_ are obser_d in thc tats exposed _othe high test substanceconccnUxtion,the same _sau= will beexami_a_from thcassig_dl=a ratsImr sexexposedto theloam-test_bm:ma_ conccntrmions. Should rcsults warrantexaminationof the lower dosage grOUl_madconduct of quantitativecwtlm_ma, scheduled reportdate andprices will be adjustedac_rdingly. The postlactlonal ovary should containprimordial and 8rowinS folliclcs as "wellas the large corporalutes of lactatima. Hislopathologir_tlexaminationmay dctcct qualitativcdcplctioa of the primordialfollicle populatioL A quanamtivccvaluation ofprimon_ follicles will be canducU:dfor Fo _ femaale_ the number oft-m, ovm-hmscction scioction andsection sample size will be stzti_'cally Spln-opriatfro"the evaluation px'_c_ureusccL ExaminationwiU include mumenstion of thenumberof Inimontialfollicles, which can be combinat with snail growing folliclcs, for compmison of ovariesofrm assigned to tretted andconla_l groups. Shlm)ine Instructions: Tngucstobe numincd histologically wilblesh_ (ambimactonclifiomt)o: --- Priacil_ In_ W. l_y Brown, D.V.M., Ph.D., ACVP v_am_,P_o_osi_ ResearchPathology Scrvicca, Inc. 438 E. But.l_ Awmue New Britain, Pcmms3dvania18901 Tdq_o_. (2_ 345-7070 Telcfax: Email: (215) 345-4326 WRBRPS_:oncentric.uet Thcrecipie_ will be notified in advanceof sample _ilnnc=aL Exygen Research Page 98 of 153 418-028:PAGE G-99 Exygen Study No.: 023-072 PROTOCOL 41 $-025 ORAL (GAVAGE) COMBINED REPEATEDDOSE TOXICITYSTUDY OF T-T/06 WITH THE REPRODUCTIOWD_ELOPM]_CI'AL TOXICITY S(:XEENINGTEST SPONSOR'S STUDY ]qUMBE_ T-T/06.I Ammldmem 1 - 17 Apri/2002 1, IXtniled Ulimeal Obs_tiom - MJlcji_d Female Rats (pal_ 10 of tim InmocoO: _me: t April2002]Dm/ledd_t_will_mtbe n_xd_t form_lmd famale rm misned m to_ mn#_. o.. TI_ dlanl_e was made _ _ data far _ of detmled clinical obs_,,atioos will be availablefrom the ms asdiF_edto the main study. 2. Feed Comum__ti'onV81u_- MaleRats andFemale Rals (page I 1 of timprotocol): [Effnctiv_lNae: 1Ai_2002) F_l_m._tattptioavtlmmwillmtimnmmd_t_ tabdatedformale_ndf,._d- ratsmlgned totoxicokin_c sampli_ ThischanScwasmadebecaasestt_dmt datafor evalual_ of feedconsumption will beavailablefromtheratssuignedto themainstn_. Any revisions made m _ fmaliz_ _mendment must be made by subsequent smadmar. Exygen Research Page 99 of 153 418-028:PAGE G- 100 Exygen Study No.: 023-072 Pmmca1411;-{_8 Ammalmm1u ..-.. 3. Es_us Cvclinst*ridMatinR (pa&c12 o f theprotocol): _t_cctivc Date: l _pril 2002] For tl_ rats usisncd to toxicokinetic sampling, cstrous cycling will be evslm_l duringthe coh_imion paiod until srm'ma_zoa are observed in sme._ of the _ controls and/oza copulatoryplug is observed/n J_, but not duringthe dosage paiod, prior to cohablt_on. This c3ama_w_ made ba:msse_t dm fi=r_ of estmus c_linl will bavm'lablc_om tlzcrats sssignat to the main study. 4. Scheduled Saczificq - ToxicokJn_c Study (page 16 oft_ pmtocor): [Xtrec_veDsm 5 Xpril2002] l_mml_ofimplmmui_aimmad_ lutza will be reconl_ O=cassa wtqlbc discanicd without futt_ avs/uaSon. ]_son ofC_nec: Th_ d_c wasmadein oed_m pinkie mo_ in_m=tion _=out_a*,,'bk toxlcit_oftb=l_s__ in pre_n=_rats. 5. ScheduledS_-i_ce - Main Studv(l_q_ 16 of thepropel): [Effective Date: 27 March2002] The mnnb_ ofimplmlxtion sites will bc recorded,ratherthan tim number of insplmmioa sites and corpora lmea will bc recorded. gsm_L_aaau: The numb_ of corpora lutezwill not be reconlzd Imcmr_ corporalulc_s at a r_pidr_e and _e not cmmtaJ oa maati.zaatwemlinll. 6. Sclzdulal Sacrifice (pa_ 19 orthe protocol): [Efl'cctivcDate: 4 April2002] The fiverfi_xneach se,lected pup win be e0tcised and thc oripmwcigh_ tccon:kd, mthcrthin fl_ liver fi'ome_a selected _ _ be coUcctal excised madthe organ wcigl_ A Any revkto_ made to tl_ _mslized mdmmt mast be made by s_sseq_t smmdmm_. Exygen Research Page 100 of 153 418-028:PAGE G-101 . Exygea Study No.: 023-072 lls_u_afi_rChan_: l_llllco1418-0111 Ammdm_ t rs_a cla_ clm'i_mtheprotocolbyremo_h_agn exmaaeo_word. Aa,u,,.a.b,,,,,,,. . ... G-.- D rr Th_.saI-L Woodsrd, D.V.M. M Mcmb_,Imlit_ioA_mlmalCare andU_ Committ_ _mButmkoff_Ph.D.,DABT, StudyMonitorand Spo_o_s CIHDate Asy revbiom msds to tkis B_lisld ammdmest mint be msde by s_qsmt smmdmmt. Exygen Research Page 101 of 153 418-028:PAGE G- 102 Exygen Study No.: 023-072 PROTOCOL418-028 ORAL (GAVAGE) COMBINED REPEATEDDOSE TOXICITYSTUDY OF %7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENINGTEST SPONSOR'SSTUDY NUMBER: T-7706.1 Amendmem2.- ! 5 July 2002 1. SafutyProcaufiol (page 3 of the pmt_l and Attachment3 paKe1 of the protocol): [EffeetiveD_. 29April 2002] A halfface respiratorwill be wom in addie_onto A glovesn,ppcopd_eeyeImXoctio,n,,s,_dfoxm/labcoatdm_g formulation _on ofd_ bulktesstubstmc This change was made to matchthe bulk test substancetext with the text locked _tl_ the_ Saf_ DataSheet 2. Study Schedule (Attachm--t I pale 2 of the protocol): [E,ffe_ve Date: 29 _ 2002] The da_esforFOB and motor activity evaluations have been extmdrd to fourdays (06 MAY 02 - 09 MAY 02) rathe_ than two days (06 MAY 02 - 07 MAY 02). _" Any revisions to this ffaudised8nteadment must be made by subtequeut amendment. Exygen Research Page 102 of 153 418-028:PAGE G-!03 Exygen Study No.: 023-072 lh_t_to14184)28 Anlendme2nt l'aS:e ReasonforClaant_: Thisc]_ml_wns madebecausmeore limeisneededtoevaluataenimalasssigned to FOB and motor activity cvalun_ons. d- " _ofR_..srch -- Asso_st_ Director_l_l_escarch Study _r _-Th_ H. Wo0d_D',_L Mmaber,Instit_oml Animal Cnrc and Use Committ_ Date John Bullmhoff,Ph.D., DABT, CIH Date Study Monitor and Sponsor'sRepresentative Any revisions to tbb baUzed mmdm_t must be rode by subsequent smeEdme_L Exygen Research Page 103 of 153 418-028:PAGE G- 104 Exygen Study No.: 023-072 PROTOCOL 418-028 ORAL (GAVAC_) COIVlBINE;RD.I_F,,ATPD.ODSE TOXICITY"STUDY OF T-T'/0W5ITH THE R.EPRODUCTIONJDEVF,.LOPM]_ITAL TOXlClTY SCREENING TEST SPONSOR'S _'UDYNUMBER: "I"-7706.1 Amendment 3 - 17 July 2002 1. Bulk Test SubstanceSmm31inaS. him_inzInstructions.Shitmin2_op..s. Caeuulm-Sectioainz - Toxicokinetic Study._chedulcd Sacrifice - Toxicokinetic Study. and Scheduled Sacrifice. (pagcs 4, 5, 12. 13, 16 and 19, rcspccttvely of the Im_m;ol) [Effective D_. 29 May 2002] Samples farpedlmmhcxmesulfonate (FFHS) m3alysisshipped to Lisa Ciemon at 3M En_ Technology and Safety Services will be resl,dppedand remaining samples, thatwere to be shipped to Lisa ' Clemea sud have not yet bern shipped, will be shipped to: Johnl_dm_yP,h.D.(PrincipaInl vcstigatar) 30511Research Drive Stale CoBcl_ Pcnnsylvoniz 16801 Tclcphonc: (814) 272-1039, ext. 122 Taed'ax: (814) 231-1580 Email: joha.flsh_com Any revisioss to this fisallz_ amendment must be made by subsequent amendment, Exygen Research Page 104 of 153 418-028:PAGE G-105 Exygen Study No.: 023-072 Pmtno141_S Amenttmem3 P'_e2 Thesesampleisnclude: Bulktestsubstancse_mplin(gpage4ofthcpTotocol) Concmtx_on and homoBcn_ty (p_,c 5 of the protocol) Stability (page 5 of the protocol) Plasmatoxicokineticsamples(pabc, i I to 12andpagc16 oftheprotocol) Plasmasamplesr,atherthanserumsmnplesw, crc_ned. Liver toxi_ldncic s_mplcs (pages 12 and 16 of the protocol) Pooled fetal sos'ran tox_ddnctlc samples Coa_s 12 and16of theprotocol) Pooled fetal liver toxicokinedc munples(p_es 13 md 16 of the protocol) Pooled pupserumtoxicokinetic samples(page19 of the protocol) PUPliver tmdcokinetic ampler (page 19 of the protocol) The analymmwiUbe Imbcouttaetedto _ by th_ Sponsor endthe Quality A.ummce Unit for Exysen Resear_ will conduct criticalphase inspections mad auditthe res_ve remits and repom accordingto the StandardOperating psocedar_ of that fadlity. Such criticalphasc impccdon rcpom and audi_reports will bc subcuiaedbythat facilityto the Study Director,RaymondG. York. Thc date of the inspections and reportsubmissionswill be incorporateidntoa QAU r4atm_mt gmcraml by Exygm P,_ch for imlusion in thefiredreportfor Protocol418-02& __. Kcamn forCharm: This chan_ was made st the requestofthc Sponsor bccausc 3M is not able to complete the fmmulatioa mml_Mswith their cummt stains. i''/,v,_rd_-I-M--'-H-/,_,Ib.a_nan'PI_D'DABT / Dittct_ofP.cscsrch D= I_y]_JsdG. Yos_.I_.D.,_)ABT Datc Asso_ DiTcctooirP._sj_h StudDyircc_ Th_ _ Woodmd, b.V.M. " Member. Insti_tional Anim_ Care and Usc C.ommiuce Date JohnButenhoff, Ph_.D.D, ABT, CIH D_c Study Monitorsnd Sponsor'sRcprcscntafivc # ._y revisions to this fiuib_ _nt.dm_t rest be male by _btequmt amettlment. Exygen Research Page 105 of 153  | 418-028:PAGE G- 106 Exygen Study No.: 023-072 PROTOCOL418-028 ORAL (C_ COMBIN]93_TED DOSETOX_CITYSTUDY OP T-7706 WITHT/_ RRPRODUCTIO_PMENTAL TOXIClTY_I_INO TSST SPON_OR__JTUDYNUMBI_ILT: -7706.1 " _meadmeat 4 - 20 l_aa'ch 2003 _._n-S_cfimdn_ -T_ SUMS. Schexlal_SaIcrifi_-Tc_fi_kin_c Study.aad_ (three _, 5, 12,13, 16 md 19,n_aectivety af the andAmeadmeat_. Itemh BulkTe_tSubstanceSamvline. _]alLialtal_ t3_me_._aiae - ToxieotieeeeSmdv- Scbednled$aatfl_e- Toxi_daet_ Study.ead $c.hede_ Sa_fice (page I of Amendmea3t) will bedoaeaccotclb_eoK_ygeaMettuxlKxM-023-071Revieioa1,eatitled "Methodaf_ for tb_De_mia_ioa oflae_am_bexaaem_om_a)m, RatL_v=,Se_una_dUdn_Rewis'1c"_. Aay _ te th_ fmaltffiedmmmdmeatmint be made by mlm_lmmt ammdme_ Exygen Research Page 106 of 153 418-028:PAGE G-107 Exygen Study No.: 023-072 t i !..... , Exygen Research Page 107 of 153 418 -028:PAGE G- 108 Exygen Study No.: 023-072 ",.,, . ' "- PROTOCODLEVIATION DeviationNumber:. 1 Date of Occurnmce: 10/14102 Page 1 of 1 Exygen'StudyNumber: QZ3-072 ProtooolNumber:. 418-028 ,. DESCRIPTION OF DEVIATION A recovew o[ 62_ for PFHSIs being accep_d for the 5000ppb _olke (0202119 SpkC) in data set 100402A. ",,, Oevlatlon Issued. _ ''\ forexamok-d_lvlalkmkm._l.SOPrev_lon_.,'m. R._ord_e:_a0,., IMPACTION STUDY o._,0:711S]o3 The 10 ppb _ 50 ppb controlmatrix Sl_.k_ Includedwith the set gave acceptabte recoveries. %., k.,.. "\. " Date Mahagefnenl Slgtta_e _._./.___ c_po._-o*" z. _ U-:tFamwR_o'rocot,_f_VtATlON.dtm_ "', , A_, " Date .....o:,/,-s/_oo-_ sx_ QAUR_ew'7'h,,)'/ 'l//s-/,._ . ".,.... _I.0_ OI_)N_L. m '_._ r..,_,__,,,_o_,u_ Exygen Research Page 108 of 153 418-028:PAGE G-109 Exygen Study No.: 023-072 RESEARCH . StateCo_teEleP,A 16801 3058ResearchDrive Fax: 814-231-1580 Phone:814-272-1039 NOTETOFILE Date: 01114/03 ExygemResearchSludy# 02,30,72 PROTOCOL#. 411F92B NOTF_ ThePFHSstocksolutionwasinadvertentlynot correctedfor salt content whenprepared.The stocksolutionwasusedto makethe fortificationand calibrationstandardsusedforthe study. Dueto the largeamountof dataassociatweldththisstudy,the concentratiomof fortification andcalibrationsotutloP_ttsted _t the raw data were left uncorrectedfor the salt content. The final PFHSvaluesrecordedwere correctedto reflect the differencein PFIt5 concentrationdue to the salt content. ThecorrecUonwasperfmmedbymuRtplyingthe final ppb valuefoundby 0.91, si_at_:' tJ o,t_: 01llq!o_ Exygen Research Jur3y1z,oo1_o Page 109 of 153 418-028:PAGE G-110 Exygen Study No.: 023-072 APPENDIX B Analytical Method: Method of Analysis for the Determination of Perfluorohexanesulfonate (PFHS), Perfluorooctanesulfonate (PFOS) and Pentadecafluorooctanoic Acid (PFOA) in Rat Liver, Serum and Urine Revision 1 (Exygen Method No. ExM-023-071 Revision 1) Exygen Research Page 110 of 153 418-028:PAGE G-111 Exygen Study No.: 023-072 MethoodfAnalysfiostrheDot=ruinaotf_ion (PIWIS), Perfluorooctanesulfonate(PFOS)and Pentadecafluorooctanoic Acid (PH3A) in Rat Liver, Scram and Urine Revision I AUTHORS JohnFlal_'ty, _ Risha, andEmily Decker November20,2002 SPONSOR 3M Medicsl Depenn_t CorpommToxicolo_ 3M Ceater, Building 220-2E-02 St Paul, MN 55144-1000 PERFORMING LABORATORY Exyge_ Research 3058R_h D_ State College. PA 16801 METHOD NUMBER ExM-023-071 Revision I TOTAL NUMBER OF pAGES 43 Exygen Research Page 111 of 153 418-028:PAGE G-112 Exygen Study No.: 023-072 _tyll_ MethodNo: E._M-O_O'/1R_m I MANAGEMENT APPROVAL a.2 "" _ kln_ger Ex_a't Research Joim L. Butevhoff SponsorRepn_ntative 3M MedicalDepartmen_t Date Toxicology .KIyLeaResearch Exygen Research PlSe 2 of 43 Page 112 of 153 418-028:PAGE G-113 Exygen Study No.: 023-072 TABLEOF CONTENTS TITLE ....... .................................... ............... ......... ...................... .................. .................. .1 MANAGEMENTAPPROVAL........................................................................................ 2 TABLE OF CONTE/CrS...........................................3............................ LIST OF TABLF_ .......................................................................................................... 4 LIST OF FIGURES .......................................................................................................... 5 1. SUMMARY ................................................................................................................. 7 2. I_{I_R/MENTAL COMIK)UNDS.............................................................................. 8 3, CHEMICALS AND SUPPLIF.S ............................................................................ 9 3.1._ ..............................................9............................ 3.2._rh.ND_S ..............................................9............................ 3.3. _ _ Strn_ ................................................................................... 9 3.4..SOLUa_ ..................................................................................................... I0 3.5. I__ o_ ST_ SoLIJ'tao_ ....................................................... I0 3.5.1. Stock solution....................................................................................... 11 3.5.2. Ftmification Solutio_ ......................................................................... 11 3.5.3. Calibral_onSlmd_ds .......................................................................... 11 4. METHOD................................................................................................................ 12 4.1. FLOWD_ ........................................................................................... 12 4.2. S_iPI.B PR_ ..................................................................................... 12 4.3. BATCllSI_ uP.................................................................................................. 13 4.4. S_lI_ _ 4.4.1. Livcr l_x_ ...................................................................................... 13 .................................................................................... 13 4.40.. _ madUrineExlractiou............................................................... 13 4.4.3. SIrE Column Coa_litiot_in8.................................................................. 14 4.5. l_dAlCnTXmON.............................................................................................. 14 4.5.1. _$ $y_m mdOl_atin_Coudfliom .................................. 14 4.5.9. Calibr_on Curve _ .............................................................. 15 4-5.3. Sample Amdysis .................................................................................. 16 4.6. Acc_'r_ CPaa'muA........................................................................................ 17 4.7. Pmu_o_u_rc_ C_'l_.m .................................................................................... 17 4.8. _ Rm_trme_r_._A_Ar,._S_.S....................................................................... 18 5. CALt2ULATIONS ................................................................................................... 18 6. SAFETY ................................................................................................................... 19 ]_.xylle_a Pai_ 3 of43 Exygen Research Page 113 of 153 418-028:PAGE G-114 Exygen Study No.: 023-072 Exyll_ M_'_xlNo:F.,v_-02..=-071R4,CIatoa LIST OF TABLES Table I. Recovery Summm7 of PHIS in RatLiver and Senun....................................... 20 Tablc2. Recovery Summary of PFO$ in RatLiver, Serumand Urine ........................... 21 Tab|c 3. Recovery Summaryof PI_A in RatI.,ive:,Scum and Urine.......................... 22 Exygen Research Page 114 of 153 418-028:PAGE G- 115 Exygen Study No.: 023-072 Ex)'lenMed_dNo:ExM-023-07_1 1 LIST OF FIGURES R_re i. Ca_Izra_onCurve forPFHS........................................................................ ?.3 l_iEu2r._ CalilEatiC_urveforPPOS .................................2.4............................ Rgure3. CalibratiCounrw forFFOA ................................2.5............................. _-e 4. RepnmenmtiwChmmatogmmof a 0.t ng/mL Standard Containieg PFHS....................................................................................... 26 Pigme 5. RepresentativeCI-atmaaWgramof a 0.1 ng/raSLtandardContainiPnFgO$...26 Figutm6, Rept'esentaliveChroraamgramof a 0.1 ng/mL StandardContainingPFOA,.27 Ftgm_7. Repnm_tativeC_aonmtogramofa0.5ng/mLSumdm'dContaininsFFHS...27 Figum 8. RepreumtadveChromatogramof a0.5 nf,/mLStandardContainin8 PFOS...28 Pi_tm 9. _ve Chmmatogramof a 0.5 n_mL StandardConudningPFOA.. 28 Figure I0. l_ve Chromatogramof a 5.0 ng/mLStandardContainingPTHS...29 l=igun=11. Representative_'am of a 5.0 ng/mL StandardContaining PFOS...29 Figure IZ Re_v Chrom=_,D'=mof a 5.0 ns/mL StandardContainingPFOA.. 30 Figure 13. RepresentativeChtomatogramof a ReagentBlank SampleAnalyzed for Pills ..... ....,..........oo............o................o...o ....... . ........ . .......... o. ....................... 30 Figure 14. Relm_ten_ve Onomatogram of a Reagent Blank SampleAnalyzed for PlUS ........................................................................................................ 31 Figure 15. _ve Chrmnatograrnof a Reagent Blank SampleAnalyzed for PPOA............................................................................................................... 31 Figme 16. Rep_ve Ouomatogram of a ControlLivez Sample Analyzed forPFHS......................................................................................................... 32 F'tst_ 17. _tative Chromatosramof a Conlxol I.,ive_Sample Analyzed for FFOS ....................................................................................................... 32 Fi_ue 18. Relxesentative Clnomatogramof a Connol Liver Sample Analyzed forFFOA......................................................................................................... 33 Figure 19. RepteaemtativeChmmamlFamof a ControSlerumSample Analyzed forPFHS ......................................................................................................... 33 Figure20. Repmlentalive Chromamgnunof a Control SerumSample Analyzed forPTOS ....................................................................................................... 34 Rgnm 21. _tative Chromatogramof a ControSlerum SampleAnalyzed for PPOA ........................................................................................................ 34 Figl_ 2.2. _tluive Chromatoga'amof a Comml Urine SampleAnalyzed for PFOS.......................................................................................................... 35 Figure23. RInea',ntativeChromamgramof a ControlUrineSample Amdyrcd for PFOA.................................................................................................... 35 Figure 24. ReprmentativCl'aomatogramof a Control Liver Sart_le Po_! at 10 ng/g with PFHS...................................................................................... 36 Exygen Research Page 115 of 153 418-028:PAGE G-116 Exygen Study No.: 023-072 Ex_ MethodNo:_23-071 _ 1 LIST OF FIGURES(continued) Figure 25. l%:ln_-nmivc Cnmmatoj_ramof a Comm! Liver Sample Fortified at 10 ng/g with PFOS ................................................................................... 36 _.gure 26. Relxtm_tative Chrtmuttogremof a Conol Liver Sample Fortified at tOng/g with PFOA............ ...................................................................... 37 Figa._ 27. _tatiw Chromatog'maof a ControlLiver SampleFortified _ SOng/g with PF[-IS............................................................................... 37 _gtl_ 28. RepresentativeChromatogrmnof a ControlLiver SamplcPoftified m 5ong/gwithPFOS .....................................3.8............................ Figure 29.R_tafive Chromatogrsomf a Control l..iveaS"ampleFortified . at50ng/g with PFOA ...............................................................................38 Pigu_ 30. _taflve Clmmu_gram of a ControlSe_umSampleFortified at 10 ng/mL with PlrtIS............................................................................ 39 Figure31. _tative Chrom_gram of a Cmmvl Scram SampleFortified at 10 ng/mL wilh PI_S....; ........................................................................... 39 Figure32. _tative Chromatogrsmof a ControlSerumSampleFortified at I0 ng/mL with PI_A ................................................................................ 40 Pigure 33. Reptr.aeatativeChromatogramof a ControlSerum SampleFortified at 50 ns/mL with PI:_S ................................................................................. 40 Rgure34. RepresentativeChmmatognunof s ControlSerum Sample Portified at 50 ng/mLwith PPOS................................................................................ 41 Ftgure35. RepresealativoChromatogramof a ControlSennn SampleFortified at 50 ng/mL with Plea ............................................................................... 41 Figure36. Relr',_ttative Chromatogramof a Control UrineSmpk Fortified at 10 ng/mLwith PFOS................................................................................. 42 l_gtt_ 37. R_ta_ve _gra.m of a ControlUrineSample Pottif_ at i0 ng/mLwith PFOA ........................................................................ 42 _guze 38. RepresentativeCiu'm,a_gram of a ControlUrine SampleFortified at 50 ns/mL with PI_S ................................................................................ 43 Pigu_ 39.RepreacnlalCihvreomatograomfa ConU_lUrineSampleFor_ed at 50 ng/n_Lwith PI_A ................................................................................ 43 Exygen Research Page 116 of 153 418-028:PAGE G-117 Exygen Study No.: 023-072 Ex_qleMn_hod No:Exlvl-023-0_71 I I. SUMMARY This _ dermis a m_hod of analysis for residues of Perfluorohexanesulfonate (FFHS), Pm'fluoronc_ (FFOS) and Pr.ntadecafluca'ooctanoic Acid k_F()A) in Rat l..ivea', Sermn md Urine. Residues of PI_-IS, PFO$ and PFOA are ex_ from each matrix with acetonitnle. The acetonitrilc extract is added to water and loaded onto a comlitioned C18 solid phase extra.on (SPE) cartridge. Analyte residues are cluted with 2 mL of methanol. Quantification of PH_S, FFOS and PFOA is accomplished by liquid ctnumatographyhmdem meat speetmme_ analy_ using multiple reaction monitoring (MRM). (LC/MS/MS) The Im31msed limit of quantitation (I.OQ; the lowest fortification specified by the method which gives adequate nscovory aecordiag to EPA guid:linm) for this method is tO nf/S (ptr_per-bilti_) each for I'FHS, _ end PFOA. The theoretical limit of detection (IX)D) will be based on the signal to noise ratio and will be at least greater than 3 times the level of noise, bated on the ir.struln=ntation system used. For all ana]ytes, the lowest analytical standard _ds to O.lugtml. This method was developed _ rat llvez, serum and urine. Typical percent recovenea standard deviations (at 10 end 50 nl/g) are shown belmv: Level (ns/s) 10 [ $0 [ Lira 115e_ :1: 9.9_ (.a=3_ 911_:1:3_f, (w.3} Levd (m_l.) 10 [ -_ Secure 108_ + 4.7_ {_3) 111_ "9'-.6% (n_3) L(eeev/se)l. 50 PFOS Recovery in all Liver F-ccefl_,ttoa level (neYmL) l_b 4-I.S_ (n=3) 50 I'FOSRamvery tn I'FOS Rcmven/tn 1ha S,mlm Rlu l.blae 93_ :t:4,7%(n_3) 120qb+ 2.1_ (n=3) _qb 1.2_ (n_3) L_vd (_s) 10 50 PPOA P._.ow_ t_ p..tLi_ 91i9_:k3.19t,(n,.3) 94% + 2,$%(_3) Lm_ (nS_L) 10 50 PPOA t_cov_yin 1tatSe_a 11,7% l.SeI, (n'.,3) 111%:14: .0% (n_) FFOA Recover/in Rat_ae 89%:17:..J%(e=3) 157%:t:2.1% (e-_3) _tative e.alibration _rves are shown in Figures 1-3. Reprmtentative chromatograms are shown in Figures 4 to 39. ._ _ Page7 of43 Exygen Research Page 117 of 153 418-028:PAGE G-118 Exygen Study No.: 023-072 _ M_od No:P_..xM-II_3_-U'/I 1 2. EXPERIMENT_, COMPOUNDS 'I'bestructu_ forPI_HS,PFOS and Pb'OAarc give0 bel,ow. Chemical Name = Molecular weisht = Pcdlum'ohcxancaulf_aate 399, as shown FFF /F_F/F F_S03" FF F PI:q-ISis supplied as tl_ potassium salt (C_13SO3qK+)m, ol_-ul_ w_ght = 438 t_OS Chemical Name Molecular weight = Perfktorooetanc_dfonatc = 499, u F FF F FF F F_S03" I'FOS is suppliedm the potassium salt (C_a_SO_'K*) molecularwcdght = 538 Chemical Name = _tadccafluarooctanoic Mo_ul_ w_t = 413, u _mwn F FF0 F "O" ]_cysar_L I_ Sof43 Exygen Research Page 118 of 153 418-028:PAGE G-119 Exygen Study No.: 023-072 ExygeaMelh_ I_: ExM-023.0I7.RevittemI 3. CHEMICALSAND SUPPLIES 3.L Cm_c.Ats Chemical Methanol(MeOH) Ammonium Acetate Water Acetonitrile Grade I..IPLC Reagent Type I HPLC Source , Ca.mBl.oNo. EM Science MX0475-1 JT Baker 0596-01 F.,xygen EM Science NA AX0145-1 Type I water = electricalresistivity, minimum of 16.67 Ml2/cm at 25 *C, fax_aa L.abconcoWaterproTM worlumeion. 3.7,. STANDAm_ Standard Perfluorolwxanesolfmmte (PPH_) Pet4l__te (PFOS) Pentadecafluorocctanoic AcidOn'oA) TCRNumber SE-036 SD-018 Lot No: 0s31et_O Purtq,(_t) 99.99 all/seiners, 84.36 straigchatin 86.9 96 Source 3M 3M Aldrich C_m 3.3. Equa_orr A_D_ ,]_lalpmenl Balance, analytical(display at least 0.0001 g) 125-mLLDPE ha:row mouth bottles Dispoaableglassmicropipets(50-100& 100-2001aL) Tissum/zer Wrist action shaker Sorvall RC 5C ptm Cenuifu_: 50 mL disposablepolypropylenecentrifuge robes 15 mL disposable polypmpylene centrifugetubes Visiprepvacuum manifold Sep PakVac 6 cc (lg) tC18 camidges (,peat# WAT 036795 2-mL cleaHrPLC vial Kit (cat # 5181-3400) ClassA pipetsand volumetricflasks Standardlab equipment(graduatedcylinders, disposable tubes etc.) Stend-aleste drop-inguard_dse #844017-400) holder(part Hypercarbdrop-insumd colunm(4 ram)(part# S44017-400) HPLCPumpO_2-10AD) _S andI-IPI_ sys_ms Supplier Mettler Nalgene Drummond(VWR) Tekn_r Burrell Scientific .Dupout VWR VWR Supclco Waters Hewlett-Packard various suppliers various suppliers Keystone Scientific Keystone Scientific Shimadzu As describedin sect/on 4.5. _tygenllesczrcb j ........... Page9 of43 ExygenReseareh Page 119 of 153 418-028:PAGE G-120 Exygen Study No.: 023-072 Exyge_Meth_ No:ExM-023-07_1evisi_I Notes: 1. 2. 3, 4. 5. In order to avoid omaminaliou, the use of dispoaable labware (_ntainers, tutm_pipex_ m=.i)shighlyrcconmzm_ct_. Teflon or Teflon-lined containers c_ equ_ment should not be used. It may be necessary to check the solvents (a_,tonitrilc, methanol) for the presence of contaminants (especially PFOA) by I.f,JMS/MS before use. Certain lotn_ have been found to be unsmtablc for use. Use dispoeabl miempipcttes or pipettes to aliquot standard solutions when preparing standardasnd samples for exlnet/ma. Equivalent materials may b_ subs_tuted for those specified in this method. 3A. SoLtmcevs (I) -2 mM ammccixma acetate in wat_ is prepared by weighing 0.154 g of ammonium acetate and dissolving in 1 L of watt. (2) Hypercazb filtered type I water is prctmred by filtering type I water ttmyagh a Hyperemb guaz.d column using a EIPLC pump at -2-3 ml./min. Before use, wash the guard cartridge with -25 mL of HPL_ grade acetonitrile, then - 2.q mL of type I water, then begin collecting the filt_,,d type I water eluate for use in the _traction, Repeat the wash after filtering -21.ofwater. Note: The aforemcofioned example ia provided for guidance, altez,native volumes my be prepared as long as the appropriate ratios of the solvent to solute are maintained. 3.5. PREPARATIONOft STANDAIIDSOLUTIONS Analytical standards axe used for three purposes: 1. Calibration Sumdmh - These standards m_ p_pm_ in methanol and are used to calibratc thc response of the dctcctor usod in the analysis. 2. Laboratory Control Spikes - These fortifications are prepared at conc.entratiop_ c_mmling to the LOQ and 5x LOQ and ate used to determine analy_cal recovery. Laberatory control spikes are prepm'ed in control matrix. 3. Matrix Spikes - These fortifications are prepared by spiking into the field samples at a known Conecntration. Matrix spikes are used to eva/_e the effect of the sample matrix on analytical rccove_ and ere prcpared _t the client'rsequest. The tma]yst may vary the absolute volumes of the standards as long as the correct proponims of solute to solvent are maintained. P_YI_s _ Pap 10 of 43 Exygen Research Page 120 of 153 418-028:PAGE G- 121 Exygen Study No.: 023-072 Exyi_ MethodNo:.KOJl-0O.3-0R"/m1_sionI 3.5.1. Stock soluaon Prep_s in_viduaI stock so]t_om at 100 _/_L for FFI_, PPOS and PFOA by weighing out I0 mg of _ anal_dcasl tandard(cot'rect_t for purity and if no:euary, salt comc_t) and dilu_ m I00 mL with methanol in _'parate 100-n'_..,volumetric flasks. 'l'be stock solutions (in 125-mi., LDPE bottle) m'e m be stm,ed in a refrigerator r, 2"12to 6"C and an: stable fora maximum periodof one yearfrom the da_ of pr_ar_on. 3.5.2. Eolian $ohaions a. Preparea mixed fortitica_ou standant at 1.0 pg/mL (lO00 ng/mL) of PFI_, PPDS madPFOA by sdding 1,0 mL of rach of the I00 pg/mL stock solutions into a 100-mLvolumetricflask madbling up to volume with methanol. b, Prepare a mixocl fortification standardat 0.1 lag/mL (100 ng/mL) of IK"I-13I,:_OS andPFOA.bydiluling I0.0 ml..of tim 1.0 pg/ml.,mixed fotl/_ation solution to 100 mL with mexhanolin a vohumetncflask. F.,aall3_: one hundredmicrd/_ g of liver or I mL of _ us/g) fortification. of th_ 0.1 _ghr,L solution spiked in_ I is equivalent to a I0 ppb (10 npJmL c_ Stc_ all fortification standardsolutions in arr,frigca'atcr(in 125-mLLDPE bottle) at 2Cto 6"C fora maximum periodof one year from the damof incpmation, Note also that additionalconcentrationsmay be preparedff necessary. 3.J.3. Calibration ..Wandardl I.C./blS/b_ _th'brationstandardsconm/ning PFfL_P,POS madPFOA am pre[mred, at 0.I, 0.2, 0.5, 1.0, 2.0 and5.0 ng/mL in methanolvia dilution of ttm0.1 lag/mL mixed fortification soimiou (section 3.5.2.b). "l'lm following is a typical e.,xamplc;additional con_utmtiopa may be pmpm._l as mmaed. InitialCone. (nghnL) 100 100 I00 5.0 2.0 1.0 Volume (mL) 5.0 2.0 1.0 10.0 10.0 lO.0 Dilut_ m (mL) 100 100 I00 I00 I00 lOO Final Conc. (nr/wJ.,). 5.0 2.0 1.0 0.5 0,.2 o.1 The standards my I_usedfura pcdod ofoneycm"(in 125-mLLDFE boulm) when sumxl mhigm'at_d(at2"C to 6"C3. ._y]l_aI_u_,h Pqcll of4] ExygenResearch Page121 of153 418-028:PAGE G- 122 Exygen Study No.: 023-072 Exyp_nMethodNo:F_M-_3-071RcvisioaI 4. METHOD 4.1. FLOWDL_G_ The flow diagram of themethodis givenbelow, followedby a detailed dcaczip_oifoenachttep. McthodFlowDiagram Wcigh appropriate amountof fiver or measure alqnopriat_voltm_ of serum_ urine (fortifysamples designmedas matrix spikr_and laboratoryc_atzol spikes) ,L Add water to livc_ for a final volume of 10 mL, add wat_ to serum and urine foxa final volumeof20_ homogenize Remove ImL andadd5mL ofACN, shake ceatafuge J, Decant supcmatantinto 35mL ofwater Load onto conditionedSPE Elutc with 2 mL mcthanol I.F_/MS/MASnal_s 4.7.. S_Qq.Z I_tOCZSS_O Forliver samples, place ftozcn samples in a food processor madhomogenize with dry ice.. Then place ssmples in containers and leave opcu in frozen stot'agc ov_rhght to allow for COz rablinmticm. Seal and place the samples in fr0zcn storasc below -10*C until time of extraction. Altea-natdy, if thcrc is an insuffi_ent amount of sample (- less than 5 8), then no processing is and the sample can be used as supplied. No uunple Im3Ce_ng hi needed for se_um and urine samples. However, frozen senun and urine samples must be allowed to completely thaw to room teml:_-atu_ bcforc usc. .l]xyRgmeaetrch Page12of 43 Exygen Research Page 122 of 153 , i 418-028:PAGE G-123 Exygen Study No.: 023-072 ExysenM_thodNo:Ev.M-0234YPT.mI 4sloa! 4.3. BATCHSETtip L A batchof samples should not contain more flum20 field samples. b. Each batch of uanples analyzed mu_ iaclud_ at least one eoaatrol(method blank using control matrix) and two max_ controls fordfied at known concentrations(typically 10 and 50 ng/gfor liven"or ng/mL for sezum and urine) to verify [nocedural recovery for thebatch. c. At least one field sample in each bat_ must also bc separatelyfortified at a known concentratmioandcarried through the _ to verify m_,ea'y. Additionalsamples in the ba_h may also be fortified if desired. d. All samples rl_qllix_dl.lp]icatcinjections. 4.4. SA_ Ex'n_CllO_ 4.4.1. /.dyerF_racaan a. Weigh 1 8 of liver sample into a 50 m]., disposablecentrifuge tube and fortify, if appropriate, Note that altca'nateweights of liver may be measureddependingon the sample tire availableforuse. b. Addwater to the sample for a final volume of I0 ml, Captightly. c. Homogenize sample using a tissuemizerfor -1 minute. d. Trmmfer 1 mL of the sample using a disposable pipette into 15 mL disposable ceutrifuge tub_. Add 5 mL of ACN and shake for -20 minuteson a wrist actionshaker. e. Centrifuge tubes at -31300 rpm for - 5 mimam. _y decant supematmatinto a 50 mL disposable centrifuge tube and add 35 mL of waist. f. Load the sample onto a conditioned SPE column (for conditioning details. see section 4.4.3.). Dir,,ard the duam. Any mudyte residues will be trappedon the SPE colunm at this point. g. P/ate with 2 mL of methanol. Collec2tmL of lutc into a graduated15 mL ceatrifugctube. h. Analyze samples usingclcctrmpmy LC3MS/MS. 4.,#.2. Seruraand Urine E,xtmclion a. Meaau_ 1 mJ..,of serum or urine sample into a 50 mL disposable centrifugetube a_adfortify, if appropriatNeo,tethatalternavtoleumes of serum and urinemay be measureddependingon sma_e sire available for tl3c. .F_tylenResemr_ Page13of 43 Exygen Research Page 123 of 153 418-028:PAGE G-124 Exygen Study No.: 023-072 EXyltebnlethodNo:ExM-023-071R=vtaiIoa b. Add wal=r to mmxplfor n firedvolume of 20 mL Cap tightly madvortex for -I minule. Tbencomint_ with #tel_ d-h in motion 4.4.1. 4.4.3. SPE ColumnConditioning Place _ unconditionedSPE columns on the vacuum manifold. Condition t_ SPE columns by passing - 10 mL of methanol through tim column followed by - 5 mL of water. Tke washes may be pulled tlnou_ tlm SPE column using vacuum at a flow rateof -I drop/me or may be allowed to pa.,,_throul_ lhc colunm unaida:LDiscardall waslms. Donor allow thecolmam todry. _. Qu_rrrAxxo_ 4.5.1. LC/M_.WIVSISystem and Openm*ingConditions MassSl_: Interface: Compute:. Software: IVl_crom_ Qu_ro Ultima t'Mictom,_) El_y (lVlica'ormm) Harvard infusion pump ('/-Ire'yarIdasmmamats),for tuning COMPAQ_omd Wodc_aticm AP200 Windows NT. Mamlynx3.3 ItPI_: H=wlctt l_kard fliP) Sea'ies1100 ripQuartump lip VacuumDelp_e_ HI' Autosampler HP ColumnOven Note: A 4 x 10 ram hyptqtmb da_ in gum'dcamidg= is attaol_! on-line after thz ptn_ valve and befort_ sample injector port to trapanyresidue contaminantstim maybe in thc moldl phaseand/cHFLC system. Column: Qcne_ Ct (Jones Chromato_.,q_y),2.1 mmx 50 ram,4p ColumnTemperature:35 C InjectioVnolm_: 15pL Mobile Phase (A): 2 mM AmmoniumAceta_ in Type I waU= Mobile Phase ('B): Methanol Tam 0.0 2.0 5.0 9.0 9.5 __A 9O 9O 10 10 0 _ E_Bag._aldmim I0 0,3 10 0.3 9O 0.3 90 0.3 100 0.3 14.0 0 100 0.3 14.5 90 I0 0.3 20.0 90 I0 0.3 ExylFm_ Pag=14of43 Exygen Research Page 124 of 153 418 -028:PAGE G- 125 Exygen Study No.: 023-072 ExyileaMethodNo:ExM-O23-UR'/IeviaioaI It may be necemty to adjust the HPLC gradient in order to optimize insmmaent I_formanee. Columnswith diffenmtdimensions (e.g. 2.1 x 30) andalso eoionme from different numufacumnl(Keyslooe Betasil Czsetc.) could be used, provided e_luiValentdnemams_hy is obtained. Ions monitored: /klZZfl._t Mode _ P]_tS Negative 399 --_ 80 PI_S Negative 499 _ 99 Pl_OA Negative 413 --_359 Appmxtma_ RetentioTnime -8.2 rain. -8.8 rain -8.6 min The retentimatlrae.,may vary, en a day to day basis, depending on the batch of mobile phate etc. Drift in retention times (up to 4 %) is act_vteble within an analyticalrun, m long as the drift continues through the entire anaiy_ andthe ztandard_axeincluded at thebeginning and endof the analyticalrim. Note: An alternativeLC/MS/MS systemmay be usedoncedemonslxal_tlo be equivalent. The ma_ _ is tunedfor eachanalyte by izff_in8 t -, 1.0 p,g/mL standardeo]utioe(at I0 I/.[Jmin,ruinganinfusionpump}via a "1" intoa sura, m of mobilephmecontaining50% methanoal nd50% 2mM ammonitma_cetatein waterat 0.2 mlJmin flow rate. Eachanalyteis initially emaefdor theparention and then tuned for the Imxluction. Once the insmmmnt is toned, the optimized pm_meten are saved as a tune file. This tune file is then used durin8 routine analy,it. 4.5.2. Calibraflon CurveProcedures a. Inject the same aliquot (between I0 to 50 pL) of each calibration standard (ransingfi_mnthe lowest level standardto the highest level pt_-pared)i,nto the I.C/MS/MS. b. Use weighted linear standardcurves for quantitation. Linear standardcurves me generated for each mudyte by linear regression using I/x weighting of peak area verms alibnttien standard concentration using Mastlyr_ (or eq.ivalmt) software system. Anycalibration standardfound to be a rtatiuical outlie_ by _ing an appropriate outlier test, may be excluded from the calculation of the calibrationcurve. However, the total number of calibration umadardslhat may be excludemdustnot exceed20_,ofthe total numberof standardsinjected. c. The correlation coefficient (11) for calibration curves generated must be _0.9925 (Rz >0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and lh_ _mdarda or thezr.levantset of tamples should be zeanelyzed .ExyleaP.ema_ch Page15of43 Ex'ygeRnesearch Page 125 of 153 418-028:PAGE G- 126 Exygen Study No.: 023-072 Typicalcatibratioucurvesfor PF_, PPOSandPI_OAcan_ fotmdin R_ 13. 4.3.3 _e AJ_z_s_ a. Inject the same Ll/quot (between I0 to 50 laL) of each standard, sample, recovery, eonln_l,etc. into the/..C/MS/MS systen_ b. Standardscorresptmdingto at _ fir= or more concentratio_ l=vr2s(starting with the LOQlevel or below) must be includedin an amal_ieal s_. c. An enti_ set of calil_mli_ _tndards ahouldbe injectod at the beginning of a _et followed by calibfatiml_andh_ intea_en_ approximatelyevery 5-10 smnples (to account for a second set of exlmcted stlmdenk). As an altm_tiv=, an cntixe1t of calibrationstandardsmay be includedat 1t beginnin8andatthe_1 of aeample set. In tither case,cah'l_-a_ standards must be the first end lastinjection in a samploset. d. The conc_atral/cmof ench aamplc/forlificafion/contml is determinedfrom the standard c_rv=, based on tl_ peak area of each snilyte. The mndud tmpom_ shouldbracket responses of the residuefoundin eachsample set. R_ults may be quanfitatedup to 10% outside the curve by extrapolation. If necessary, diluttehemmpl_ to give a response within the stmdazd curve rlmge. e. Poaification recov_es fallingwithin 70 to 1309t are considcred acceptable. f. Samples mustbe r,tvt'edrefxigeratedbetween 2C to 6*C until analysis. g. Sampl_ in which eiflu= no pea]_ are detected or peaks less than the lowest concenntion of the calil_ation sm_ are detected at the coacsponding _malytez_-_en_ontime will be n_po_x[ _ biD (not det_). Samples in which peaks am detected .. the cocrmpond_nganaly_ n_cu_ou time that arc leas than_ LO_ and _zeatcat-han or equalto thc lowest conccntzafionof the Mbn_ standardsw/ll be reportedas NQ (not qmmflfisble), The analysis pedmmed dung the method devclOl_aCaitncluded fortific=tiom at 10 and 50 ng/g of PFHS in na liver, lOand 50 ng/mL of PPHS in serm_, 10and 50 ng/g of FFOS madPFOA in ratliver and 10 and 50 ng/mL of P_3S andFIK)A in serumandurine. Typicalchromatogramacan be found in Figur_ 4-39. Px, ylcnP.escazch Pap 16o(43 Exygen Research Page 126 of 153 418-028 :PAGE G- 127 Exygen Study No.: 023-072 ExygmMeth_No:1_tM.023.07_1.-.mt I 4,6. ACCtFrAN_ The following critermiuast be met to rostra: the pt_smacoefPl_d_,Pt_OSand PFOA: I. Thech_m_ogram mustshawa peakofa daughteixonat80araufrom _t of399ainuforPF'HS,a daughteironat99 ainufroma p_a_notf 499 ainu for PFOS, and a daugh_ ion at 359 ainu from a paxc_ of 413 ainu forPFOA. 2. MetbedblanksmustnotcmatahaannIyl_atlcvedsgreatertlIufmtheLOQ. a blank _ntaim the anal_e at levels s_,ater than I0 ng/mL, thea a new blmak_maple mint I_ oblzin_ md tbe _atir_-t m_t be _ 3. P_..ovl_i_ of cclztl_l spikes madmatrix spikes (if any) must 1_ between 70-130% of lheir known vnlues. If a control spike fails outside the acceptable limils, the entire set of _mples should be re-extracted. Any malr_ spike outside 70-130% should be evaluated by the analyst to if re-extraction/s wan'anted. 4. Any calibrationstanda_ found to be a statisticaloutlier by ruing the Huge Eaur Test, may be excluded fnzn the calculalion of the cah'brmioncurve. Howsve_, the total number of cah'lmaion_ that could bc cxc]udcd must not cxcecd 20_ of tbe totMnumberof stnndmdsinjected. 5. The eorrelalion codtieient (R) for cnlibrnfioncurves generated mutt be Z,0.9925(R2 _>0.985). If caI/brafionresults fall outside these limittsh,en appropriatsetepsmustbetakenm adjusitmminentcpe_, andthe stand.I_sthereleamtsetof_ples should be nnmlyzod, 6. Retentiotnimesbctwelm_ and samplesmustnotdrifmtore than :t: 4 % within an ano/ytical run. If zet,ention _ d_ exceeds this lirait withinan analyticalrunthcn tbe sct must be reanalyzed. 4.7. Pn_,om_cz Cm'rmuA The following two c_i*a_lamust be pcrCmtm_ once al a system suitability W.st, be.forethe comme_e_ent of analysis,whea using an instnm_nURionset-up d_at iresnot becn used for this method. Run a standardsolution m LC3MS/MS correspondingto the estimated LOQ (I0 ng/mL) in matrixand obtain a signal w noise ratio forthn anadytetransitionof at least 9:1, comparedto a reageatblank. If Ibis critmioncanaot be met, olximize and clumgc in,_tmumt operating panunete_ (or increase the injeetkm volume, if eppropam). P.xyfp_Research Palp_17of43 Exygen Research Page 127 of 153 418-028:PAGE G-128 Exygen Study No.: 023-072 EzylenMethodNo"E_J_-_I P_w I Second Criterion: Run a set of standardsof five or more concentrationlevels, from at or below the LOQ, up to the highest coaeentralion level to be included in the analysis. Generate a calibrationcurve for the anallae and obtain a linear regrmsion with a coefficient of demtmination fR2) of at least 0.985 for the analym. Oaee this criterion is met, aamplenmay be analyzedwith standardsintm'spersed. 4.8. _ _ 1,Oi A_YstS Oneperson can take a set of 20 samplest_-_b thesample prepm-ationprocedm_ in appfoximatoly4 hours. The I.C/ME/MS analysis of riteset (containing20 field sample_, I matrix blank, 2 laboratory conlrol spikes, I matrix spike and 12 standardinjections) will take appmximamly14 hours. S. CALCULATIONS a. Use Equation 1 to calculate the amount of maaly_efound (in ng/mL, _ on peak area) using tim ste_'d curve (1/x wedghted linear regression parameters)generau:dby the Masslynx softwmpmgrar_ Eouafion I: Anal3afeound(ng/mL):_ slope xDI:xaliquofta:tor DF = faet_ by which the fma] volume was diluted, if necessary. Aliquot factor=-10 for liver, 20 for serumandurine b. For samples foaified with known mnountsof analytprior to _traction, use Equation 2 to calculatethe percentreonvcty. Eauation 2: Recovery (%) : [ totalanalytofotmd(ng/mL) - mudytefonndin conla_l (ng/mL,)]xl00 analytoadded(ng/mL) Note: Submmtanalyte found in omatrol(ns/mL) from malyte found (ng/mL), if ng/mL in control is greate_thanLOQ. E_yIFaResearch Pa_ IBof43 Exygen Research Page 128 of 153 418-028:PAGE G- 129 Exygen Study No.: 023-072 EarYl__e_ N_ EJ_M-ID3_7_1 1 c. LIneEquation 3 to calculatc thc amountof analy'tcfound ('mppb) Anslym_nd (nl_llm"nll/mL=) rmudvtfeomM, _'ng/mLx'P_VIrmL'_ szmplewzi_ts(tZ._ _ '*ol'mn(eml.) PV = final volume Par tcpmang purposes, samples in which cithcr no peaks arc dcw.ctcd _ pcaks lcu thmathe lowest co_:ntntion of the callbralion standardsmt dct_qcd at tim _g mal_ _nnion tim_ win bc _-por_ a r_O (not _n:_t). Ssunp_ in which peaks axedetected at the contslmnding msly_ n_entica time that amless thmatim LOQ and IFcatm"than or equalto Oaclowest conccntrationof tim calibrationstandm-dswill btcponnd as NQ (not quantifiable). Tlz mmlyst should hum thc matm'ia]sa._ty data sheets _ all stnndan:Isand r_$nmts bcf_ lm'formiug dais mctlaod, UN uuive_d im_._mtiot_ whoa handlingstandards and rcagcnts, including working in fume hoods and wearing laboratocryoats,safety_asse,,madglovm. .Exy_ _ Exygen Research Pip 19ot"43 Page 129 of 153 418-028:PAGE G-130 Exygen Study No.: 023-072 ExYI_nbl_axl/_. Ex.,_..Q23-07l1_ion ! TableI. RecoverySummaryofPFI_ inRatlaver andSerum Recovery Summary of PFH$ In Pat Uver O=OlSa4 Spk S 0_01se4 spk c 10 10 " ' Avm'age: Standw'dDevi_on: 110 12s 115 '- llJi Sam_,!O . _ 0201664 Spk D _-_01684SpkE o_SpkF , Added(_) SO 50 so Avmage: 8blndlrd Deviation: _,,.t _ q_co_w _) ,, 101 98 118 _L5 Recovery Summary of PFHS in Pat Serum 0_01682 Spk B 10 10_ _spkc ,o ,lo 81_dard Deviation: 4.7 0L_01_2Spk D 02016e2 Spk E _1682 Spk F 50 107 50 104 50 I 122 Avelap." 111 Oevlstlon: 9JS .]Bxyi_]Rmcarch ExygenResearch Pap:20 of43 Page 130 of 153 418-028:PAGE G-131 Exygen Study No.: 023-072 Kstyl_nl_lbod No: ExM_I l_vilio_ 1 Table 2. Recovery Summaryof PFOSin Rat Liver, Serum andUrine Recovery Summary of PFOS In Rat Liver 0201M4 8meA 0_01M4 8pkB _mgmSf,kC _o 1o 10 Average: Stamk._Oevbdlon: 88 95 , 1,o5 s6 S.S hmpls ID 0201M4 SpkD OL,_m4 8_ E ml_ 8_,F AMIm Added(rig/g) 50 S) 5o Awmme: 8tabard Detlmtkm: lq,mecd Recovery (%| Im n a7 8| 1.6 Recovery Summary of PFOS In Rat Serum hmple ID 020168281)kA GgOt68gSpk8 AnelyhAdded(ng_L) 10 lO 8mnde_l_ Pen:antRecowm/(%| im 85 O9 as u hmpe :) O201682Spk o 0201S8281_ E ,G201_ 8pk F AnsI_,, Added(ng/mL} sO GO 50 &wmge: Stsnd_rOl eWd_m: F_r_,nt P.eco_ry ('/.) 1'to. S_ 121 120 2.1 Recovm'y Summsry of PF08 in Rst Udne i_.,0 _1M2 8pitA O2Ola2 8pkB (_01682 8_kC ,_,y, A_,d_.ll_ lO 1o 10 , , Awm_ S_tdsrd Dsvlstk)m ,_._._.,.:_-, _ 98 O9 91 _ ' 4.? O_Olm _ S _ __ sO 6o .. 8_rd .&wrNle Dovtstlon: 7e _, . "/I 1.2 _zyll_'* Rmum:h Pq_e21 or43 Exygen Research Page 131 of 153 418-028:PAGE G-132 Exygen Study No.: 023-072 F_ty_ M_I_! 1_: F-_tM_I Rnvi_a t Table 3. RecoverySummaryof PFOA in Rat Liver, Serumand Urine Rm:a,vm'y Summary of PFOA In Pat Liver _ 8empleID o2o,,leeS4 _kA I)20'1664SpkB 0201M4 8_ C An_/le Aclded(ng/g) 10 I0 10 A_ IIw.lml Dovlall_. Pementt_xmv_y (%) N I01 99 , M _ 02O1884EpkE 0201M48pkF SO 50 Am Oevfetkm: , 94 94 ;LS Recovery Summary of PFOA In Rat Serum 0201M28_pAk n_116828pka 0_01U2 SpkC 10 lo 10 A'_le; Stm_hudI)mh_m: 118 117 115 ' 117 1J _01682 8pkO SO 107 0201_ SgkE SO 112 020_ 8pkF 50 116 Avemp: 111 StandmdDevtdon: 4.0 Recovery Summary of PFOA in Rat Urine h,,y. ,o 0201662SpkA 0201(182S_ B 0201e82_ C, _ _ I"l_l ) ,_"m _ _) 10 86 10 91 10 , Average: 8indwd Deviation: 89 ' , 89 2..6 C;mlee2Spk0 so mt 020_2 Sl_ E SO 118 0201682GpkF , 50 ,,, 85 Almnlge: 87 StandardOevfsti_: 2.1 .EXylleaRese.mgh PNge22 of 43 Exygen Research Page 132 of 153 418-028:PAGE G-133 ' Exygen Study No.: 023-072 _>',_s Method No: F_d-4)'2..1-_ 1 Re_o I FigurIe. Calibm_mCurveforPFH$ Exygcn Research Page 133 of 153 418-028:PAGE G- 134 Exygen Study No.: 023-072 _tylen Metl_d No: F.._I-I_3-071 _ I Figure2. CalibratlonCurvefor PFOS Compound2 name:PFOS _nt Of13eros: 0.9970O9 CaJibmfio_nlrve: 3211.30 "x+ 48.9985 Flesponmtype.E" xternal_ Ares Curvetype:linear, Origin:Ex_ude,Wel_Jno: 1/_ Ai bans: None . II _e 1.621,4. Con o'_ .............. 1'o 1'_ zov ..... 2'.s'....... _.o 3._i .... 4' '...0.. ,5i .... s. .Exyl_ P._sesn_ Exygen Research psp 24 of 43 Page I34 of 153 418-028:PAGE G-135 Exygen Study No.: 023-072 Exy_,_ _ No: E_v_-02__ i _ l Figure 3. Calibration Curve for IrFOA _nd 1 namo:PFOA Go_tiaent of De_'rr/maian: O._S_tS C_ curve:31270.9 x+ 3675.38 _ma_nae ty_: E_,_ud S_ Curve(ype:Linear,OdWn:Exaude,We491_nglh: _ A_stramr 1.66e5- .... 0.0 i .... 0.5 1"1 " " _ .... 1.0 1.6 _ .... 2.0 i _' - i .... 2.5 3.0 i " L . ., i .... 3.5 4.0 i - _ . . _ n_?t_ 4.5 5.0 - , ii i i Exygen Research Page 135 of 153 418-028:PAGE G-136 Exygen Study No.: 023-072 Exygta blethod No: gx1602._071R_isi_i I Figure 4. RepresentativeChroma_ Cont.ainl_ PFHS :ola,/m4. L't aNaL M'I'_ 110 of a 0.1 nV,,/mLStandard _.,I.,l_,..Wm 'uq,km_N mo I lu_l tlalL ,t ................ " ,.m " .m " _ ..J-.tA. .L._l.._,_,. ' A.bO-' Km* _ 7_" _l..-d itt" _ ......... '_ a_JLat Lr,.. ' i_" _.m Figura_5. RepresentativeChromnlo_'am of a 0.1 ng/mL Standard tI "_" .....*_. Exygen Research Page 136 of 153 418-028:PAGE G-137 Exygen Study No.: 023-072 F._yl_M'end_l No:Es.M-I_3-071ReviIsiaa Fiware 6. RepresentativeChromatogramof a 0.1 ug/mL Standard ContainingPFOA l._MaltM drf 1.80 2.00 _Lm ,LOO LN ann ?_ JLO0 9.00 _ 11.80 Figure 7. RepresentativeChromatogramof a 0.5 ng/mL Standard ContainingPFHS _T,'MWU8#7 l ._ ,, , kt _ . .... ' 1_d ' t;- ' _b," _ ",i._ ' t_ ""i_, ' tU "tin * ._m ' ,_ ' I/*T*M _xyl_a Research P_t_ 27 of 43 Exygen Research Page 137 of 153 418-028:PAGE G-138 , Exygen Study No.: 023-072 Exyipm lg'h_dmd No: F.,xM-023-O71 Rnviaion I Figure 8. RepresentativeChromatogramof a 0.5 ng/mL,Standard 1_1i,I ._ :_-_ tk_.:_o ,. _,." tm --" - _r.m -tbuT-a _ " ,/m" ._o ' n. Figure 9. Representative Chromatogram of a 0.5 ng/mL Standard Containing PFOA .E,.tygcm. Research Exygen Research l_age 23 of 43 Page 138 of 153 418-028:PAGE G- 139 Exygen Study No.: 023-072 _xyg, m Me_Jd No: KxM-023-071 _ I Figure 10. Representative Chromtogram of a 5,0 ng/mL Standard ContainingPIllS LC_.US 01. Figure 11. RepresentativeChromatognm of a S,Ong/mL Standard Conta/ningPFOS Lr.Ma_ o '. ,_ ,_.-.,._.. ,_ - .,.. ,_ .',_, .,.. i, L ,_,' ;,_. -,,-- Exylp_ ]_mu_ _%q_ 29 of 43 Exygen Research Page 139 of 153 418-028:PAGE G- 140 Exygen Study No.: 023-072 F-.xXI_a Method No:/_tM-4_3.-071 RcYimiou 1 Figure 12. RepresmmtaUveChronmtogram of a S.0ng/mL Standard ContainingPFOA ml, u qdm. _ Ilzmol_lm Im Ot_l_l rio _ lq_i _a_,zla zrx, 1_-M u_m_mu _ ttt_ tl It_l_ |_/lll 413_lR I <_o L_ ' +_o " ..U " +_, ' o,_ ' 'T_.-" 0_o'' "u+ " '_ ' ,_;_, ..... 'nma, Figure13. RepresentativeChromatogramof a Reagent Blank Sample Analyzedfor PFHS nmqm_ Imm_A glz01A._ ik_IMe_, tm Z_-Um _ l._mmm _ _m lammm_B. t_ te 4m _w te ILlS _ tl_ ._dm ll_lemm_ PII_ 30o_43 Exygen Research Page 140 of 153 418-028:PAGE G-141 Exygen Study No.: 023-072 Ex_l_m Mn_a_l No:. Ktl01_l _,cviaca I Figure 14. Kepresentative Chroamtogramof'a Reagent Blank Sample Analyzedfor PFOS tqml_t Imvt A _dt_,_ 10_l_qt IW _ "W _.m " _ '_ " _-"_.m " _ ' Tm " _ " _ " u " .:m " '_ Figure 15. RepresentativeChromatogramof a ReagentBlankSample Analyzedfor PFOA IlllpllHIlllB A _l_ Illl II1_ IhQ gG-_W-ImJm:Nd_ | 1 I_II, IMUl m' WIIMWll _umls D- in ._xyl_u l_esetrch Exygen Rese,areh l_qlc 31 0(43 Page 141 of 153 418-028 :PAGE G- 142 Exygen Study No.: 023-072 ExylpmMee_l No:K_tM-023-0R"e7v1iskmI Figure 16. RepresentativeChromatogramof a ControlLiver Sample Analyzedfor PFItS t.cmmu_ _ tN Figure 17. RepresentativeChromatogramof a ControlLiverSample Analyzedfor PFOS _m4 uw# m_t I_Mg_Ito_ Imam M_asu_ _m_ MP_#_I_ IMP,4 .K_y_aRmear_ Exygen Research Page32 of43 Page 142 of 153 418-028:PAGE G-143 Exygen Study No.: 023-072 Method No: KxM-(_I-071 _ I Figure 18. RepresentativeChromatogramof a Control Liver Sample Analyzedfor FFOA Imot_t Liv_ m A _11mlN _ln, _ m.kel :N --_'_ Iln, MRM J_ Cm_lfl IRI- Figure 19. RepresentativeChromatogramof a ControlSerumSample Analyzedfor PFHS Immll_l Ilam_m Iltamk & mr_,'4a am tt_ a,_ tad s.w a4-Jmt410R _ LG4M_Me tl _QII _EIb MS_U am Lm _1 L Exygnn IteP.arch [ _ l_.p 33 of 43 Exygen Research Page 143 of 153 418-028:PAGE G-144 Exygen Study No.: 023-072 F-xy[InaM_lalxl No" K_tM-02.3-071 ll_dlim I Figur2e0. RepresentatCihvmenmtogmm ofaControSlerumSample AnalyzefdorPFOS _ _imltA Bli_I_Ul C11:411:t Figure21. RepresentativeC_nromatogramof a ControlSerum Sample Analyzed for PFOA Olg_nm8t ramIIImdlA el_idamlA._ Ira(.loing, l4_ _ _4&auO t.PJ,IJ_ ap_r U_e_d I GDmWtIS- ............. _J am am _ tim am 7_ " too ' .- . .'_fanm am lo_m "i1_o " .r_yL_IIResem_h Pq_ 34 _43 Exygen Research Page 144 of 153 418-028:PAGE G-145 Exygen Study No.: 023-072 _ Metl_d No: ExM-G2.3-071Rcvlsioa 1 Figure 22, RepresentativeChromatogramof a ControlUrine Sample Analyzedfor PFOS mMQm Uet_ EmQ A 0_s_.m4 am/h6s,I_ U_t_lam _ t._hmmmet u_ _ _tCs_m_ ESdkN m Atom lqgure 23. Representative Cln-omtogram of a ControlUrine Sample Analyzedfor PFOA _.ms _ Imnt_ s m._e,419R im_._ I#.JMMn _ _ 41J_b.I_k, i t,m _ _ _ IkU IkW T.m _ ItM IIUsO 1i_110" Exygen Research Page 145 of 153 418-028:PAGE G- 146 Exygen Study No.: 023-072 Exy_ Metbcxl No:. _LM-QI3-O'/1 _ 1 Figure 24. Representative Chrmmtogramof a ControlLiver Sample Fortifiedat I0 ng/g withPFHS _M4 t_vet _A. WW_ idt_m_-_ 8mg_. 2_} _,_R4sm s-'2_0o t.ot_qm p _qkt a I c_m_toQ. _t_, _m A.I Figure7.5. RepresentativeChromatogramof a ControlLiver Sample Fortifiedat I0 ng/gwith PFOS aR_l.Jkwr _ toplm WU_lll Im IIIl_h_l e_e.aml s_m_t L_..JM_N6W IdW mlICI,_ II- ..... _ u,_; _ _ :-u, _ ,._: ,_._:_,,2_ _'- .]_ylpm _ Pap 36 of 43 Exygen Research Page 146 of 153 418-028 :PAGE G- 147 Exygen Study No.: 023-072 EzYlP_ Me/hod No: EzM-4Y)._-0?I P._._.s_on 1 Figure 26. RepresentativeChromatol_'amof a ControlLiver Sample Fortified at I0 ng/gwith PFOA ClQmIULlv_l_tA SOgq_ llnml_s_sl lu_a_ M-duhlaR 2_lt-_n _Sw,a| I_t4mds U- Figure27. RepresentativeChromatogramof a ControlLiver Sample Fortified at SO_g/gwithPFHS MI0V|MUbm SpkL mt_b M!1_116 k _ M MS.,Jw,t_rm,_mt_m:l_m R _10Mm_MM- aLNI|s gll7p.mR u _ss * I.m " _ ......M.e... *,_ L_ _ y_,.A.i_ ' S_; ....I_ iS" -* tl_r_ Exygen Research Page 147 of 153 418-028:PAGE G-148 Exygen Study No.: 023-072 Ex)_ea Me_cd No: F..xl_14)_-O'/l l_'_i,_ ! Figure28. RepresentativeChromatogramof a Control Liver Sample Fortified at $0 ng/g with PFOS tim ' _im" Xm _ _" _ L_'" T_" tm " _m * u " _'_ " r_ Figure29. RepresentativeChromatogramof a Control LiverSample Fortifiedat 50 np./gwith PFOA umss4 _ sl_ o, m i,_ _ t_lz L_uamm _ _ 41Ss, llm ILO I_B lm L_ LR Lie ?_ F.xyi_ ll.ctet_h Palle 38 of 43 Exygen Research Page 148 of 153 418-028:PAGE G-149 Exygen Study No.: 023-072 Exyi_ Med_ No: KxM-02.t_I Rz,4tiou l Figure 30. RepresentativeChromatOl_-_aof a ControlSerum Sample Fortilled at I0 nghnL with ulnul llm_n&_ I,tI inplP lllllalbl_IB _ lira,_ II_I_ IIIIIcNI: t._Imnm _ lJ_/,f I O_,I Ill- 1.Udl ! / Figure 31. RepresentativeChramatogram of,, ControlSerumSmnple Forl/lied at 10luq_/mwLith _OS al_ll_ IIm_m I1_ Ik to iq_ ImlB_-lu liraIIt_ I_1 II_IM.aW m-lSal7 I_Mwl _mJJil. 7_ ' i_ 'itS-' -_ ' _ " tit * ui ' _ "_ ' nit " _',_" i_" __t=. Exygen Research Page 149 of 153 418-028:PAGE G-150 Exygen Study No.: 023-072 Kxyl_nMethodNo: ExM-023-071Revision I _gur_ 32. Representative Chromatogramof a Control Serum Sample Fortified at 10 ng/mL with PFOA mmamlmvm;N & W_ gaqu_a,_ _ I1_ _ m._mR la:lm._ I_a mzf Cl_m_ i_ I1 1 _m," 41_1_me,_ Figure33. RepresentativeChromatogramof a ControlSerumSample Fortifiedat 50 ng/mLwith PFHS _f_M8 grt _xyl;ImRm_urt:h Exygen Research Page40 of 43 Page 150 of 153 418-028:PAGE G-151 Exyge_a Study No.: 023-072 34. Rep_ve _t_m ota _ FoXed at 50 n_aJ_ with PFOS Serum Sample _ms_Ms_ ) -- 3S. Represm_tl_ ChromatolFam_ a ControlSerum Smnl_ Fortlned at 50 l_,/mL wlth PFOA ........ _k_" m " a_ "aii "_ " a_ " is " t_ i_'_ _ _" Ex-yg_ _h Page 151 of 153 418-028:PAGE G-152 Exygen Study No.: 023-072 _r'/llen Idelhod 1'_Io:]_M-023-071 Revision I Figure 36. RepresentativeChrmnatol_m of a Control Urine Sample Fortified at 10ny/mLwith P]FOS mm_l_t& tm wm_ m_amma IV:aka kCmLqm4_ _. mi_t.rN_o " i_, " s.m * _ " 4dm " _ "i din " "_.m," _ "m " iKm ",h_ "_'-- Figure37. RepresentatCihvreomatograomfaControUlrineSample FortifiaetdI0ng/mLwithPFOA mmaasI/er_osL m_ 8m_B&,1,11m i_IsR m./_4mwm_kse Lc_/mln st IRA_ | CMAmlIB- l.k _ 44o _m, lira lm itm itm _ a,n_a " .Klylcn _:a:a:ch Pasz424_43 Exygen Research Page 152 of 153 418-028 :PAGE G- 153 Exygen Study No.: 023-072 L'lethod No: F.xM-1_3..071 Revixiou I Flgure 38, Representative Chroumtogramof a Control Urine Sample Fortified at SOug/mb withPFOS. m_ _nsS_ O.m_ m Ik_i'Ve,N. m,_.aum ha| C_w_s_w Fisure 39. RepresenlativeChromatogramof a ControlUrine Sample Fortified at S0 nr./mLwlth PFOA guqmua_l _ _N_ m,_Jutm, m_ mBT .P ._cyFa ]Rmmrch PaI_ 43 of 43 Exygen _h Page 153 of 153 APPENDIX H TEMPERATURE AND RELATIVE HUMIDITY REPORTS ARGUS 418-028:PAGE H-1 Temperature and Relative Humidity Report Location: Room 05 Protocol Number: 418-028 Range of Dates: 26-Mar-2002 14:20 to 10-Jun-2002 08:59 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64F to79F 77 1818,25 1817 Relative Humidity 30% to 70% 77 181825 1817 Mean (+ SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 69.2 71.0 69.2 66.0 1817 0 0 (_+0.7) (100.0) (0.0) (0.0) 54.6 64.5 55.3 36.4 1817 0 0 (+ 4.4) (100.0) (0.0) (0.0) Report Generated: 10-Jun-2002at 13:26 COMMENTS: REVIEWED BY: l,/n._ ._. _j_ DATE: (_-!0-o APPENDIX I POS1TIVE CONTROL DATA 418-028:PAGE I-1 Historical Control Data ThisFunctional ObservatiBoantterSytandarOdperatinPgroceduraendStudies conducted to document the training and competency of the technical staff and Motor Activity Negative Control Data and Positive Control Data are available atthe Testing F_'_ility. Page 1 418-028:PAGE I-2 Summary Information for Functional Observation Battery StudyNumber- Title In-Life Start Test Substance DosageInformation Number of m_ mL_ Dosages 012-006- Validation of Funclieml Observatioeal Battery and Motor A_ivity Measm_ Using Positive Test Subelances 12/89 acrylamide 50 1 7 physostign_e 0.75 1.5 1 DDT 75 1 ! 012-014- Ne_3tnxicityEvalnation of Positive Control $_ in CrI:CDBR VAF/Plus Rats 9/91 acrylamide 40 1 9 1DPN 200 1 3 _ltbaryi 75 5 1 DDT 75 5 1 _iadimefon 200 5 I 0124)15 - Neurotoxicity Evaluation of DDT in CrI:C[_BR VAF/Flus Ra_ 3/92 DDT 75 1 1 012-017- Nemotoxi_ty Evaluation of Positive Co_xol Sebsta_es in Crl:CD@BR VAF/Plus@ Rats 5/92 a_damide 40 1 9 IDPH 200 1 3 _'baryl DDT 40 5 1 75 1 ! d-amphetamine 4.0 1 ! 012-022- Ne_rotoxidtyEvaluation of Carbaxyi in CrI:CD@BR VAF/Plu_ Rats 10/92 carbaryl 40, 200 5 1 012-.031 - Neumtoxicity Evaluation of Positive Control Substances in C'rI:CD_BR VAF/Ples Rats 7/93 ac_clamide 45 1 10 IDPN 250 1 4 4o 5 z DDT 75 1 1 d-amphetamine 4 1 I Page 2 [[ [ 012-056- Neurotoxicity Evahmtion of Positive Control Substme_ in CrI:CD_BR VAF/Plus Rats [ 11/95 a_rylamide IDPN DDT 45 1 250 1 ,to t 75 1 d-amphetamine 4.0 ] 012-075 - Ne_--otoxieity Evaluation of Positive Control Substances in CYI:CD_BR VAF/Plus_ Rats 3/98 aerylamide 30 1 trimethyltin MK401 8 1 0.3 1 mbaryl DDT 100 4 100 2 012-081 - Neurotoxicity l_valuation of Positive Conlrol Substances in CrL'CD_BR VAF/PI_ Rats ll/01 aclylmnide 30 1 IDPN 250 1 d-amphetamine 40 1 loo 4 DDT I00 2 i 418-028:PAGE I-3 10 5 t ! ! 17 1 1 1 1 l0 1 1 l 5 Page 3 418-028:PAGE I-4 Summary Information for Motor Activity In-Life Study- Title 012-011 - TheAssessment of Motor Activity in Neonatal and Adult Rodmts using Passive InfraredSmsors Start 5/91 012-014- NourotoxicityEvaluationof Positive Control Substancesin CrI:CD_BR VAF/Plus Rats 9/91 0124)16 - Motor A_-fivityEvaluafiomin CrL-CI_BR VAF/Pim Rala Administm_ Chlor_mazine and dAmphetamine(Poeifive ConerolStedy) 012-058 - Nemotoxkity Evaluationof Positive ControlSubstances in Crl:CDOBR VAF/Plus_ Rats 4/96 j ii i i i m i Test Substance Dosage Information Numberof m_,g ml./k$ Dosages d-amphetamine 0.75, 1.5, 4 1 I chlorpromazinc I, 2, 4 1 I acrylamide 40 1 9 IDPN 200 1 3 75 5 I DDT 75 5 1 triadimegon 200 5 ! d-amphetamine 0.5, 1, 4 1 1 chlorpromazlne 1, 2, 4 1 1 a_/iamide 45 ! 10 d-amphe_ueine 0.75 1 1 _imeOayltin 8 1 1 MK-801 10 i 1 i ii Page 4 APPENDIX J HISTOPATHOLOGY REPORT 418-028:PAGE J-1 RESEARCH PATHOLOGY SERVICES, INC. 438 East Butler Avenue, New Britain, PA 18901 Phone: 215-345-7070 Fax: 215-345-4326 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCEENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT SUBMITTED TO: Raymond G. York, Ph.D., D.A.B.T. Argus Research 905 Sheehy Drive Horsham, PA 19044 SUBMITTED BY: w." By row,,, Veterinary Pathologist July 3, 2003 418-028:PAGE J-2 11 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT TABLE OF CONTENTS REPORT Paqe Method .......................................................................................................................1 Results .......................................................................................................................3 Summary....................................................................................................................5 Quality Assurance Unit Statement .............................................................................6 Good Laboratory Practice Compliance Statement .....................................................7 TABLE 1. Incidence and Degree of Severity of Histomorphologic Observations....................8 APPENDICES I. Histomorphologic Observations ...............................................................I-1 to 1-12 Key to Histomorphologic Observations ...............................................................!-1 Tables I-1. Histomorphologic Observations - Group I Male Rats ...............................I-2 I-2. Histomorphologic Observations - Group II Male Rats ..............................I-4 I-3. Histomorphologic Observations - Group III Male Rats .............................I-5 I-4. Histomorphologic Observations - Group IV Male Rats.............................I-6 I-5. Histomorphologic Observations - Group V Male Rats ..............................I-7 I-6. Histomorphologic Observations - Group I Female Rats ...........................I-9 I-7. Histomorphologic Observations - Group V Female Rats........................I-11 II. Individual Animal Gross and Histomorphology Data ..............................I1-1to 11-70 II1. Histomorphologic Observations in the Liver - F1 Generation Pups .......II1-1to 111-2 Tables II1-1. Histomorphologic Observations in the Liver- Group I F1 Generation Pups ...............................................................................111-2 111-2. Histomorphologic Observations in the Liver- Group V F1 Generation Pups ...............................................................................111-3 418-028:PAGE J-3 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT METHOD Microscopic examination was made of the specified tissues from 50 adult male and 20 adult female CrI:CD(SD)IGSBR VAF/PIusrats from five groups in an oral (gavage) combined repeated dose toxicity study of T-7706 (PFHS) with the reproduction/developmental toxicity screening test. In addition, microscopic examination was made of the liver of 10 pups in each of the control and high dosage groups of the F1 Generation. A brief outline of the study design showing the dose group identification, number of rats per sex per group, and dosage levels of the control and test substances are shown below. DOSAGE GROUP I NUMBER OF RATS PER SEXa 15 + 3c DOSAGE (mg/kg/day) 0 CONCENTRATIONb (mg/mL) 0 DOSE VOLUME (mL/kg) 10 II 15 + 3c 0.3 0.03 10 III 15 + 3c 1 0.1 10 IV 15 + 3c 3 0.3 10 V 15 + 3c 10 1 10 _l'en male md ten female rats of Groups I and V were assigned for histopathologic evaluation. b'rhetest substancewas consideredto be 100% activefor the purposeof dosagecalculations. CThreeadditionalratsper sex per dosagegroupwere assignedto toxicokineticsamplecollection. Male rats were given the test substance once daily beginning14 days before a cohabitationperiod that consistedof a maximumof 14 days. Dosingcontinued throughthe day before sacrifice,after completionof a cohabitationp.eriod,after a minimumof 42 days of administration.Female ratswere given the test substance once daily beginning14 days before a cohabitationperiodthat consistedof a maximum of 14 days. Dosingcontinuedthroughthe day before scheduledsacrifice(Day 22 of lactation). Dosages were adjusteddaily for body weight changesand were given at approximatelythe same time each day. The firstday of dosingwas desig- Research PathologyServices,Inc. -1 418-028:PAGE J-4 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT nated as Day 1 of the study. Pups were sacrificed and necropsied on Day 22 postpartum. All rats were necropsied and the specified tissues were collected and placed in 10% neutral buffered formalin for fixation. The testes were fixed in Bouin's solution for 48 to 96 hours and then retained in 10% neutral buffered formalin. The in-life portion of the study, necropsies, and recording of the gross necropsy observations were performed by the staff of Argus Research, Horsham, PA. The tissue processing, microscopic slide preparation and histopathologic evaluation were performed by Research Pathology Services, Inc. The tissues specified for microscopic evaluation from 10 male and 10 female rats of Groups I and V included: brain, duodenum, jejunum, ileum, cecum, colon, rectum, Peyer's patch, lung, submandibular and mediastinal lymph nodes, sciatic nerve, stomach, kidneys, spleen, thymus, trachea, urinary bladder, testes, epididymides, seminal vesicles, coagulating gland, prostate, spinal cord (cervical, lumbar and thoracic), liver, adrenal glands, heart, thyroid, parathyroid, uterus, bone marrow (sternum), ovaries, uterus, vagina, mammary gland (female rats) and all other tissues with gross changes. In addition, the liver and thyroid of 10 male and 10 female rats in each of the intermediate dosage groups were examined. The liver of 9 or 10 pups F1 generation pups from each of the 10 selected control and high dosage female rats were also examined. Representative samples of these tissues were routinely processed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin for microscopic evaluation. In addition, sections of the testes from the control and high dosage group male rats were stained with periodic acid-Schiff (PAS) reaction and examined. The study was initiated on June 24, 2002 and completed on July 3, 2003. Upon completion of the project, all raw data (remaining wet tissue, paraffin blocks, microscopic slides and histology records) will be returned to Argus Research for archiving. ResearchPathologyServices,Inc. -2 418-028:PAGE J-5 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT RESULTS The type, incidence and degree of severity of the histomorphologic changes in the specified tissues for the male and female rats are presented in Table 1. The microscopic observations in each rat are summarized in tabular form in Appendix I (Tables I-1 to I-7). A key to the histomorphologic observations precedes Table I-1. The gross necropsy observations, detailed descriptions of the microscopic observations, and a correlation of the microscopic findings with the gross changes in these rats, when applicable, are contained in Appendix I1. The histomorphologic observations in the liver of the F1 Generation pups of the control and high dosage groups are summarized in tabular form in Appendix III (Tables II1-1and 111-2).A key to the histomorphologic observations is included on Tables II1-1and 111-2. No treatment-related microscopic changes were observed in any of the male rats given 0.3 or 1 mg/kg/day of the test substance or in female rats given 10 mg/kg/day of T-7706. Treatment-related microscopic changes were observed in the liver and thyroid gland of male rats of the 3 and 10 mg/kg/day dosage groups. The treatment-related microscopic change in the liver consisted of minimal to moderate enlargement (hypertrophy) of centrilobular hepatocytes (Table 1). The affected hepatocytes were enlarged with an increased amount of dense eosinophilic granular cytoplasm. The treatment-related effect in the thyroid gland consisted of an increased incidence of male rats of the 3 and 10 mg/kg/day dosage groups with hypertrophy (enlargement) of follicular cells and hyperplasia (increased follicular cells and small follicles) in male rats (Table 1). Although the incidence in Group IV was minimally increased over the controls, the hypertrophy and hyperplasia in this group of rats could have been associated with the liver-cell changes. These microscopic changes in the liver and thyroid are consistent with the known effects of compounds that cause microsomal enzyme induction where the hepatocellular hypertrophy results in a compensatory hypertrophy and hyperplasia of ResearchPathologyServices,Inc. -3 418-028:PAGE J-6 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT the thyroid due to increased plasma turnover of thyroxine and an associated stimulation of thyroid-stimulating hormone in rats. 1 There were no compound-related microscopic changes observed in the liver of the F1 generation pups from the dams given 10 mg/kg/day of the test substance. There were a few other microscopic changes observed in the various organs and tissues which were considered to have occurred spontaneously and to be incidental and unrelated to compound administration. The type, incidence and severity of these changes were not influenced by compound administration. These changes also are listed in the attached histomorphology tables. 1SandersJ, .E.,EigenbergD, .A.,BrachtL, ..J.,Wang,W.R.,andvanZwieten,M.J.,ThyroidandLiver TrophicChangesin RatsSecondartyo LiverMicrosomaEl nzymeInductionCausedbyan Experi- mentalLeukotrienAentagonis(Lt -649,923)T, oxicologayndPharmacolog9y5, 378-387(1988) Research PathologyServices,Inc. -4 418-028:PAGE J-7 | ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT SUMMARY Microscopic examination was made of the specified tissues from five groups of male and female Crl:CD(SD)IGS BR VAF/Plus rats used in an oral (gavage) combined repeated dose toxicity study of T-7706 with the reproduction/developmental toxicity screening test. The five groups of rats had been given the vehicle (aqueous 0.5% carboxymethylcellulose), or 0.3, 1, 3 or 10 mg/kg/day of T-7706, orally by gavage, once daily for the protocol-specified number of days. In addition, the liver was examined microscopically from 9 or 10 pups of the F1 generation from 10 dams each of the control and high dosage groups. No treatment-related microscopic changes were observed in female rats given 10 mg/kg/day of T-7706 or in male rats given 0.3 or 1 mg/kg/day of T-7706. Treatment-related microscopic changes were observed in the liver and thyroid of male rats of the 3 and 10 mg/kg/day dosage groups. The treatment-related change in the liver consisted of minimal to moderate hypertrophy of centrilobular hepatocytes and the effect in the thyroid was an increased incidence of male rats with a compensatory hypertrophy and hyperplasia of the follicular epithelium in these groups of male rats. No treatment-related microscopic changes were observed in the liver of the F1 generation pups from dams given 10 mg/kg/day of T-7706. All other microscopic changes were considered to be spontaneous in origin and not treatment-related. The type, incidence or severity of these changes were not consideredto be influencedby administrationof thetest substance. Research PathologyServices,Inc. -5 418-028:PAGE J-8 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTALTOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT QUALITY ASSURANCE UNIT STATEMENT All aspects of the tissue processing, microscopicslide preparation, histopathologicevaluation and report preparationfor the study listedabove have been performedaccordingto the Standard OperatingProceduresof ResearchPathologyServices, Inc. and were audited in accordancewith the proceduresestablished by the Quality AssuranceUnit of Research PathologyServices, Inc. in compliancewith the Good Laboratory Practiceregulationsspecifiedin the protocol. OrganisatiofonrEconomCicooperatioanndDevelopme(n1t996).OECDGuidelinfeor TestingofChemicalsS.ection4, No.422: CombineRd epeateDd oseToxicityStudywiththeReproduction/DevelopmTeonxtaiclityScreeninTgest,adopted22 March1996. OrganisatiofonrEconomCicooperatioanndDevelopme(n1t998).TheReviseOd ECDPrincipleosfGoodLaboratoPryractices [C(97)186/Final]. USFoodandDrugAdministrationG.oodLaboratorPyracticeRegulationFs:inalRule.21CFRPart58. JapanesMe inistryofHealthandWelfare(1997).GoodLaboratorPyracticeStandardforSafetyStudiesonDrugsM, HWOrdinance Numbe2r 1,March26,1997. Quality assurance unit study-based inspectionswere performed as shown below. There were no devia- tions from the protocol, standard operating procedures and/or appropriate good laboratory practice regulations noted dudng the conduct of the study. The summary report of QA inspections is included in the final report sub- miffed to the study director on July 3, 2003. Dates of Dates Reported to Date Reported Inspection Study Phase Management to StudyDirector 12/16102 12/16/02 01/07/03 01/09/03 01111/03 01113/03 01/13/03 01131103 01/31/03 02/21103 02/24103 02/26103 02/27103 02/28103 07/03/03 Master Schedule Pre-initiation Trimming Embedding Microtomy Staining Organization& Review Histopathology Data Entry Data Verification Data Processing Report Preparation Pre-submissionAudit Draftreport Final report 12/31102 12/31/02 01130/03 01/31103 01131/03 01/31103 01/27103 01/31103 02/27103 02/27103 02/27103 02/27/03 02/27103 02/28103 07/03/03 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 02/28103 07/03103 ResearchPathologyServices, Inc. Karer_ W. Harkins, BS Quality Assurance Unit _ Date -6 418-028:PAGE J-9 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT All aspects of the above-referenced study performed by Research Pathology Services, Inc. were conducted according to: OrganisatiofonrEconomiCc ooperatioanndDevelopme(n1t996).OECDGuidelinfeor TestinogfChemicalsS.ection4,No.422: CombineRdepeateDd oseToxicityStudywiththeReproduction/DevelopmTeonxtaicl ityScreeninTgesta, dopted22March1996. OrganisatiofnorEconomiCc ooperatioanndDevelopme(n1t998).TheReviseOd ECDPrincipleosfGoodLaboratorPyraclJces [C(97)186/Final]. USFoodandDrugAdministrationG.oodLaboratorPyracticeRegulationFs:inalRule2. 1CFRPart58. JapanesMe inistryof HealthandWelfare(1997).GoodLaboratorPy racticeStandardforSafetyStudiesonDrugsM, HWOrdinance Numbe2r 1,March26,1997. No deviationswere noted that had any significant impact on the validity of the study. W_fR. ay _rown, DVM, PhD, DACVP Veterina_ Pathologist Date Research PathologyServices,Inc. -7 418-028:PAGE J-10 ORAL(GAVAGE)COMBINEDREPUTED DOSETOXICITYSTUDYOF T-7706 WITHTHE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINTGEST PROTOCOLNUMBER418-028 SPONSOR'STUDYNUMBERT-7706.1 TABLE1 IncidencaendDegreeof Severityof HistomorphologOibcservations DoseGroup: Sex: Numberof Animasl/Group: ADRENALGLANDS: NO. E_MINED NO. NORMAL I II III IV V M M MM M 10 10 10 10 10 10 0 0 0 10 70 00 9 -degeneratiocny,stic,cortex,mult_focal mild [0] [0] [0] [0] [0] 0 000 0 -hypertrophy/vacuolatcioornt,ex,multifocal mild [0] [0] [0] [0] [0] 00 00 0 -necrosisc,ortex,focal mild -vacuolatiocno,rtex,diffuse minimal mild [0] [0] [0] [0] [0] 00 00 0 [3] [0] [0] [0] [1] 2 000 0 1 000 1 -vacuolatiozno,naglomerulosa mild BONEMARROW(STERNUM): NO. EXAMINED NO. NORMAL [0] [0] [0] [0] [0] 0 00 00 10 0 0 0 10 10 0 0 0 10 BRAIN: NO.E-XAMNIED NO. NORMAL CECUM: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 8 000 9 -inflammatiomnu,cosa,chronic mild moderate [2] [0] [0] [0] [11 0 000 I 2 000 0 COAGULATINGGLAND: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 COLON: NO. EXAMINED NO. NORMAL 10 0 0 0 10 9 0 0 0 10 -inflammatiomnu,cosa,chronic mild DUODENUM: NO. EXAMINED NO. NORMAL EPIDIDYMIDES: NO. EXAMINED NO. NORMAL [1] [0] [0] [0] [0] 10 00 0 10 0 0 0 10 10 0 0 0 10 i0 0 0 1 i0 6 00 09 [ ] : Totalincidencoef specifieldesion,allgrades. IV FF 10 10 10 10 87 [0] [1] 01 [0] [I] 01 [0] [I] 01 [0] [0] 00 00 [2] [0] 20 10 10 10 10 10 10 10 10 10 10 8 10 [2] [0] 10 I0 10 10 10 10 [0] [0] 00 10 10 10 10 -8 418-028:PAGE J-11 ORAL(GAVAGE)COMBINEDREPEATEDOSETOXICITYSTUDYOF T-7706 WITHTHE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINTGEST PROTOCOLNUMBER418-028 SPONSOR'SSTUDYNUMBERT-7706.1 TABLEi (Continued) IncidencaendDegreeof Severityof HistomorphologOibcservations DoseGroup: Sex: Numberof Animals/Group: EPIDIDYMIDE(SContinued): I II Ill IV V M MM M M 10 10 10 10 10 -exfoliatesdpermatogeniccells moderate [0] [0] [0] [1] [0] 0 0 01 0 -hypospermia marked -infiltratiomno,nonuclear-celflo,cal minimal HEART: NO. EXAMINED NO. NORMAL -inflammatiocnh.ronic,focal minimal mild ILEUM: NO. EXAMINED NO. NORMAL JEJUNUM: NO. EXAMINED NO. NORMAL -diverticulum [0] [0] [0] [1] [0] 00 0 10 [4] [0] [0] [0] [1] 40 0 0 1 10 0 0 0 10 80 0 09 [2] [0] [0] [0] [1] i0 0 0 I 100 0 0 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 9 00 00 1 KIDNEYS'. NO. EXAMINED NO. NORMAL -basophilia/degeneratcioornt,icaltubules. focal/multifocal minimal -cyst(s)m.edulla -dilatatiopne.lvis minimal mild -infiltratiomno.nonuclear-celflo,cal/multifocal minimal -mineralizatiomnu.ltifocal minimal 10 3 0 0 10 7i 00 7 [0] [0] [0] [0] [1] 00 0 0i 1 00 02 [0] [2] [0] [0] [0] 0 i0 00 0 I 000 [2] [0] [0] [0] [2] 2 0 00 2 [0] [0] [0] [0] [0] 00 00 0 IV FF 10 10 10 10 10 10 [0] [0] 00 00 iO 10 10 10 10 10 10 10 00 10 10 98 [0] [0] 00 00 [0] [0] 00 00 [0] [0] 00 [0] [i] 0i [ ] = Totalincidencoef specifieldesion,allgrades. -9 418-028:PAGE J-12 11 ORAL(GAVAGE)COMBINEDREPEATEDDOSETOXICITYSTUDYOF T-7706 WITHTHE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINTGEST PROTOCOLNUMBER418-028 SPONSOR'STUDYNUMBERT-7706.1 TABLE1 (Continued) IncidencaendDegreeof Severityof HistomorphologOibcservations DoseGroup: Sex: Numberof Animals/Group: I II Ill IV V M M MM M 10 10 10 10 10 KIDNEYS(Continue:d) -nephritisc,hronic,focal minimal LIVER: NO. EXA_41NED NO. NORMAL [0] [0] [0] [0] [I] 0 0 OO i 10 10 10 10 10 23 20 0 -hypertrophhye.patocellulacre,ntrilobular minimal mild moderate [0] [0] [0] [9] [10] 00 08 4 00 0 15 00 00 1 .inflammatiocnh,ronic,focal/multifocal minimal mild [B] [6] [7] [6] [5] 54 76 5 32 0 00 -lipidosist,ension,focal -necrosisf,ocal minimal 02 O 00 [0] [0] [0] [0] [0] 0 00 0 0 -vacuolatiobne,patocellulamri,dzonal mild [0] [0] [1] [0] [0] 0 0 100 -vacuolatiohne.patocellulamru,ltifocal minimal mild LUNG: NO. EX#}NIEID NO. NORMAL [0] [1] [I] [0] [0] 0 10 0 0 0 0 1O 0 10 0 0 0 10 10 0 0 0 10 -infiltratiopno.lymorphonuclear, perivascaurl/perbironchail mild [0] [0] [0] [0] [0] 00 00 0 .inflammatioinn,terstitialf,ocal minimal [0] [0] [0] [0] [0] 00 00 O -macrophageasl,veoli,focal minimal [0] [0] [O] [O] [0] 0 0O 0O LYMPHNODE_MEDIA_STI_L: NO. EXAMINED NO. NORMAL B 0 0 O 10 ? O0 OB -congestion minimal [0] [0] [0] [0] [0] 0 00 0 0 IV FF 10 10 [I] [1] 1I 10 10 87 [0] [0] 00 00 00 [2] [i] 2i 00 00 [0] [2] 02 [0] [0] 00 [0] [0] 00 00 10 10 99 [I] [0] 10 [0] [i] 01 [0] [I] 01 10 10 7 10 [2] [0] 20 [ ] = Totalincidencoef specifieldesion,allgrades. -10 418-028:PAGE J-13 OP_AL(GAVAGEC)OMBINEDREPEATEDOSETOXICITYSTUDYOF T-7706 WITHTHE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINTGEST PROTOCOLNUMBER418-028 SPONSOR'SSTUDYNUMBERT-7706.1 TABLE1 (Continued) IncidencaendDegreeof Severityof HistomorphologOibcservations DoseGroup: Sex: Numberof Animals/Group: I II Ill IV V M MM MM 10 10 10 10 10 LYMPHNODE,MEDIASTINA(LContinued): -hyperplasilay,mphocytic/plasmacytic mild [0] [0] [0] [0] [0] 0 00 00 -mastocytosis minimal [1] [0] [0] [0] [2] i0 00 2 LYMPHNODE.SUBMANDIBULAR: NO. EXAMINED NO. N_L 10 0 0 0 10 S 0 004 -hyperplasilay,mphocytic/plasmacytic minimal mild moderate VdV4MARGYLAND: NO. EXAMINED NO. NORMAL [5] [0] [0] [0] [6] 10 00 2 2 0 00 0 2 0 004 -necrosisf,ocal minimal NERVE,SCIATIC: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 OVARIES: EXAMINED NO. NORMAL -cyst(s)i,ntraovarian PARATHYROID: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 PEYER'SPATCH: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 g -mineralizatiofno,cal minimal PROSTATE: NO. EXAMINED NO. NORMAL [0] [0] [0] [0] [1] 00 00 1 10 0 0 0 10 60 0 06 IV FF 10 10 [1] [0] I0 [1] [0] I0 10 9 43 [6] [6] 10 35 2I 10 10 10 9 [0] [1] 0I 10 9 10 9 10 10 9 10 10 10 10 10 10 10 10 10 10 [0] [0] 00 [ ] = Totalincidencoef specifieldesion,allgrades. -11 418-028:PAGE J-14 ORAL(GAVAGE)COMBINEDREPEATEDDOSETOXICITYSTUDYOF T-7706 WITHTHE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINTGEST PROTOCOLNUMBER41B-OZB SPONSOR'STUDYNUMBERT-7706.1 TABLE1 (Continued) IncidencaendDegreeof Severityof HistomorphologOibcservations DoseGroup: Sex: Numberof Animals/Group: I II Ill IV V M M MM M 10 10 10 10 10 PROSTATE(Continue:d) -atrophyf,ocal/mutlifocal minimal moderate [1] [0] [0] [0] [2] 10 0 0I 0 0 00 i -inflammatiocnh,ronic,focal/multifocal minimal RECTUM: NO. EXAMINED NO. NORMAL [3] [0] [0] [0] [3] 3 0 0 03 10 0 0 0 10 90 O 09 -inflammatiomnu,cosa,chronic minimal mild SEMINALVESICLES: NO. EXAMINED NO.NORMAL [1] [0] [0] [0] [i] 00 0 0i 1 00 00 10 0 0 0 10 10 0 0 0 10 SKIN _GROSSLESION) NO. EXAMINED NO. NORMAL 0 00 0 0 0 00 00 -dermatitisc,hronic,focBl minimal mild SPINALCORD CERVICAL: NO. EXAMINED NO. NORMAL SPINALCORD LUMBAR: NO. EXAMINED NO. NORMAL SPINALCORD,THORACIC: NO. EXAMINED NO. NORMAL SPLEEN: NO.EXAMINED NO. NORMAL [0] [0] [0] [0] [0] 0 0 00 0 0 0 00 0 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 10 0 0 0 10 -atrophy mild [0] [0] [0] [0] [Of 00 00 0 STOMACH: _INED NO. NORMAL 10 0 0 0 10 B 0 00 9 [ ] - Totalincidencoef specifieldesion,allgrades. IV FF 10 10 10 10 10 1D [0] [0] 00 00 11 00 [I] [I] 01 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 9 [0] [1] 01 10 10 76 -12 418-028:PAGE J-15 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH mE REPRODUCTION/DEVELOPMENTTAOLXICITYSCREENINGTEST PROTOCOLNUMBER 41B-O2B SPONSOR'SSTUDY NUMBERT-7706.1 TABLE 1 (Continued) Incidenceand Degreeof Severityof HistomorphologicObservations Dose Group: Sex: Numberof Animals/Group: STOMACH(Continued): I II Ill IV V M M M M M 10 10 10 10 10 -dilatation,mucosal glands minimal [I] [0] [0] [0] [0] 1 0 0 0 0 -edema/inflammations,ubmucosa,forestomach mild [I] [0] [0] [0] [1] 1 0 0 0 1 -edema/inflammations.ubmucosa,glandulararea minimal mild moderate [2] [0] [0] [0] [0] 1 0 0 0 0 0 0 0 0 0 i 0 0 0 0 -erosion(s),glandularmucosa moderate TESTIS (H&E): NO. E_MINED NO. NORMAL [0] [0] [0] [0] [0] 0 0 0 0 0 10 0 0 1 10 I0 0 0 0 10 -degeneration.multifocal mild TESTIS (PAS): NO. EXAC_INED NO. NORMAL [0] [0] [0] [1] [0] 0 0 0 1 0 10 0 0 1 10 10 0 0 0 10 -degenerationm,ultifocal mild THYMUS: NO, EXAMINED NO. NORMAL -atrophy moderate marked THYROID: NO. EXAMINED NO. NORMAL [0] [0] [0] [1] [0] 0 0 0 1 0 10 0 0 0 10 10 0 0 0 10 [0] [0] [0] [0] [0] 0 0 0 0 0 0 0 0 0 0 I0 10 10 10 I0 5 3 5 5 3 -hypertrophy/hyperplasifao,llicularepithelium minimal mild moderate [2] [3] [2] [4] [7] 0 I I 2 0 2 2 1 2 3 0 0 0 0 4 -ultimobranchiablody/cyst TRACHEA: NO. EXAMINED NO. NORMAL 3 4 4 2 2 10 0 0 0 10 6 0 0 0 9 [ ] : Total incidenceof specifiedlesion,all grades. I V F F 10 10 [1] [i] 1 1 [0] [1] 0 1 [2] [3] 0 0 1 3 1 0 [0] [1] 0 I 10 10 9 9 [I] [I] I 0 0 I 10 10 5 5 [0] [0] 0 0 0 O 0 0 5 5 10 10 10 9 -13 418-028:PAGE J-16 ORAL(GAVAGE)COMBINEDREPEATEDOSETOXICITYSTUDYOF T-7706 WITHTHE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINTGEST PROTOCOLNUMBER418-028 SPONSOR'SSTUDYNUMBERT-7706.1 TABLE1 (Continued) IncidencaendDegreeof Severityof HistomorphologOibcservations DoseGroup: Sex: Numberof Animals/Group: TRACHEA(Continued): -inflammatiocnh,ronic,focal minimal mild I II Ill IV V M MM MM 10 10 10 10 10 [4] [0] [0] [0] [i] 2 0 00 0 2 0 00 1 URINARYBLADDER: NO. EXAMINED NO. NORMAL 10 0 0 0 10 10 0 0 0 10 -inflammatiocnh,ronic,focal minimal UTERUS: NO. EXAMINED NO. NORMAL -distentionl,umen mild moderate [0] [0] [0] [0] [0] 00 0 00 -macrophagepsi.gmented minimal mild moderate marked VAGINA: NO. EXAMINED NO. NORMAL IV FF 10 10 [0] [1] 01 00 9 10 8 10 [I] [0] i0 10 10 01 [0] [2] 0i 01 [10] [8] i0 22 75 01 10 10 10 10 [ ] = Totalincidencoef specifieldesion,allgrades. -14 418-028:PAGE J-17 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT APPENDIX I HISTOMORPHOLOGIC OBSERVATIONS 418-028:PAGE J-18 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT KEY TO HISTOMORPHOLOGIC OBSERVATIONS - = No change (not remarkable, within normal histologic limits or indicated change not present). * = Tissue not available (specified tissue missing, insufficient tissue in plane of section, artifact precludes evaluation, or specified tissue not present in section). < > = Microscopic finding(s) in tissue(s) with gross observation(s). <-> = Within normal limits [no microscopic change(s) to correlate with the gross observation(s)]. P --- Indicated change or lesion present 1 = Minimal degree or amount of indicated change or lesion. 2 = Mild degree or amount of indicated change or lesion. 3 = Moderate degree or amount of indicated change or lesion. 4 = Marked degree or amount of indicated change or lesion. SS = Scheduled Sacrifice I-1 418-028:PAGE J-19 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 41B-O2B SPONSOR'SSTUDY NUMBER T-7706 1 TABLE I-I Dose Group: Animal Number: Sex: Death Type: ADRENAL GLANDS: -vacuolation.cortex,diffuse HistomorphologiOcbservations I I I I I I I I I I 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 M M M M M M M M M M SS SS SS SS SS SS SS SS SS SS - I - i - 2 - - BONE MARROW (STERNUM): BRAIN: CECUM: -inflammation.mucosa, chronic COAGULATINGGLAND: COLON: -inflammation.mucosa, chronic DUODENUM: EPIDIDYMIDES: -infiltration.mononuclear-cell,focal ...... ...... - 3 - 3 - - - ..... - - 2 - - - ...... I - I I i HEART; -inflammation.chronic, focal ILEUM: JEJUNUM: KIDNEYS: -cyst(s-Tm.edulla -infiltration.mononuclear-cell. focal/multifocal LIVER: -inflammation.chronic, focal/multifocal LUNG: I - - 2 - ..... ...... - <-> - - P - <-> I - - - I I 1 2 1 1 2 2 - i ...... LYMPH MODE, MEDIASTINAL: -mastocytosis LYMPHNODEfSUBMANDIBULAR: -hyperplasia, lymphocytic/plasmacyttc NERVE,SCIATIC: PARATHYROID: PEYER'SPATCH: PROSTATE: -atrophy.focal/multlfocal 1 - - 3 - 2 ...... ...... ...... - - * - * - 1 3 2 - - - 1 - - - I-2 418-028:PAGE J-20 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 TABLE I-i (Continued) HistemorphologicObservations Dose Group: Animal Number: Sex: Death Type: I I I I I I I I I I 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 M M M M M M M M M M SS SS SS SS SS SS SS SS SS SS PROSTATE (Continued): -inflammation,chronic,focal/multifocal I - - 1 - I RECTUM: -inflammation,mucosa, chronic SEMINAL VESICLES: - - 2 - - - ...... SPINAL CORD, CERVICAL: ...... SPINAL CORDr LUMBAR: ....... SPINAL CORDr THORACIC: SPLEEN: ....... ...... STOMACH: -dilatation,mucosal glands ..... -edema/inflammations,ubmucosa,forestomach ..... -edema/inflammations,ubmucosa,glandular area .... 1 - 2 - 3 1 - TESTIS (H&E): ...... TESTIS (PAS): THYMUS: THYROID: -hypertrophy/hyperplasiaf,ollicular epithelium -ultimobranchialbody/cyst TRACHEA: -inflammation,chronic, focal URINARYBLADDER: ...... ..... 2 - P - 2 ...... .... - - - P 2 - p - 1 - 2 - 1 - I-3 418-028:PAGE J-21 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-77D6 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T7706.I TABLE I-2 HistomorphologicObservations Dose Group: Animal Number: Sex: Death Type: II II II II II II II II II II 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 M M M M M M M M M M SS SS SS SS SS SS SS SS SS SS KIDNEYS: -dilatation,pelvis <-> <2> <I> LIVER: -infla-_'-'-mmcahtrioonni,c,focal/multifocal 2 2 i I - 1 1 -lipidosis,tension, focal - -vacuolation,hepatocellular,multifocal P .... - P - - 1 - - THYROID: -hypertrophy/hyperplasiaf,ollicular epithelium -ultimobranchialbody/cyst - - 2 - P P - - 1 - 2 - P P - I-4 418-028:PAGE J-22 | ORAL (GAVAGE)COMBINED REPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.I TABLE I-3 HistomorphologicObservations Dose Group: Animal Nun_ber: Sex: Death Type: LIVER: -inflammation,chronic, focal/multifocal -vacuolation,hepatocellular,midzonal -vacuolation,hepatocellular,multifocal THYROID: -hypertrophy/hyperplasiaf,ollicular epithelium -ultimobranchialbody/cyst III Ill III Ill III III Ill Ill Ill Ill 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 M M M M M M M M M M SS SS SS SS SS SS SS SS SS SS - I 1 I I 1 1 1 ...... 2 - - - 2 - - - - I - - 2 - - P P P P - I-5 418-028:PAGE J-23 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 41B-028 SPONSOR'SSTUDY NUMBER T-7706.1 TABLE I-4 HistomorphologiOcbservations Dose 6roup: Animal Number: Sex: Death Tvoe: EPIDIDYMIDES: -exfoliatedspermatogeniccells -hypospermia IV IV IV IV IV IV IV IV IV IV 19100 19103 19104 19118 19133 19141 19142 19144 1914B 19150 M M M M M M M M M M SS SS SS S$ SS SS SS SS SS SS <3> <4> LIVER: -hypertrophy,hepatocellular,centrilobular 1 1 2 I I 1 1 - I i -inflammation,chronic,focal/multfiocal 1 1 I - i 1 I - TESTIS (H&E): -degeneration,multifocal <2> TESTIS (PAS}: -degeneration,multifocal <2> THYROID: -hypertrophy/hyperplasiaf,ollicular epithelium 1 -ultimobranchialbody/cyst - - - 1 - - - P P - 2 2 - - I-6 418-028:PAGE J-24 ORAL (BAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINGTEST PROTOCONLUMBE4R18-028 SPONSOR'SSTUDYNUMBERT-7706.1 TABLE I-5 HistomorphologicObservations Dose Group: Animal Number: Sex: Death Type: ADRENALGLANDS: -vacuolation,cortex, diffuse V V V V V V V V V V 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 M M M M M M M M M M SS SS SS SS SS SS SS SS SS SS - - 2 - - BONE MARROW (STERNUM): BRAIN: ..... ...... CECUM: -inflammation,mucosa, chronic - - 2 - COAGULATINGBLAND: ..... COLON: ..... DUODENUM: ..... EPIDIDYMIDES: -infiltration,mononuclear-cell,focal - 1 .... HEART: -inflammation,chronic, focal ..... 1 ILEUM: ..... JEJUNUM: -diverticulum <P> ...... KIDNEYS: -basophilia/degenerationc,ortical tubules, focal/multifocal i ..... -cyst(s),medulla P - P .... -infiltration,mononuclear-cell, focal/multifocal I - - - -nephritis,chronic, focal - - 1 .... I - LIVER: -hypertrophy,hepatocellular,centrilobular 2 1 2 2 3 2 1 2 1 1 -inflammation,chronic, focal/multifocal I - I I I - I LUNG." ...... LYMPH NODE. MEDIASTINAL: -mastocytosis - i I .... LYMPH NODE, SUBMANDIBULAR: -hyperplasia,lymphocytic/plasmacytic 1 3 - - 1 3 3 3 NERVE. SCIATIC: ...... PARATHYROID: PEYER'S PATCH: -mineralization,focal ...... 1 ...... I-7 418-028:PAGE J-25 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-77OB WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 TABLE I-5 (Continued) HistomorphologiOcbservations Dose Broup: Animal Number: Sex: Death Type: V V V V V V V V V V 19111 19117 19124 1912B 19127 19128 19140 19145 19149 19152 M M M M M M M M M M SS SS SS SS SS SS SS SS SS SS PROSTATE: -atrophy,focal/multifocal - - - 3 - 1 -inflammation,chronic,focal/multifocal 1 - 1 - - I RECTUM: -inflammation,mucosa, chronic SEMINALVESICLES: .... ...... 1 - SPINAL CORDf CERVICAL: SPINAL CORD, LUMBAR: SPINAL CORDf THORACIC: SPLEEN: ...... ..... ...... ...... TOMACH: -edema/inflammations,ubmucosa,forestomach - 2 .... TESTIS (H&E): TESTIS (PAS}: THYMUS: .... ...... ...... _HYROID: -hypertrophy/hyperplasiaf,ollicular epithelium -ultimobranchialbody/cyst _RACHEA: -inflammation,chronic, focal URINARYBLADDER: 3 - 2 - 2 3 3 - 3 2 P .... P - .... ....... 2 - I-8 418-028:PAGE J-26 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOL NUMBER418-028 SPONSOR'SSTUDY NUMBER T-7706.1 TABLE I-6 Dose Group: Animal Number: Sex: Death Type: ADRENAL GLANDS: -vacuolation,zona glomerulosa HistomorphologlcObservations I I I I I I I I I I 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 F F F F F F F F F F SS SS SS SS SS SS SS SS SS SS - - - 2 2 - BONE MARROW (STERNUM): BRAIN: CECUM: -inflammation,mucosa, chronic ...... ..... ..... 3 2 COLON.' DUODENUM: ..... ...... HEART: ...... ILEUM." JEJUNUM: KIDNEYS: -nephritis,chronic, focal LIVER: -inflammation,chronic, focal/multifocal ..... ...... .... - 1 .... 1 - 1 LUNG._.__ -infiltration,polymorphonuclear, perivascular/peribronchial - - 2 - - - LYMPH NODE, MEDIASTINAL: -congestion -hyperplasia,lymphocytic/plasmacytic -mastooytosis LYMPH NODE, SUBMANDIBULAR: -hyperplasia,lymphocytic/plasmacytic MAMMARY6LAND: .... - 1 - 1 - 2 - ..... .... 3 - I - 2 - 3 1 - 2 2 NERVE,SCIATIC: OVARIES: -cyst(s), Intraovarian PARATHYROID: PEYER'SPATCH: ....... - - - P - - - ....... ....... RECTUM: SKIN (6ROSS LESION): -dermatitis,chronic, focal ....... <2> I-9 418-028:PAGE J-27 | ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITY STUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTTAOLXICITY SCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 TABLE I-6 (Continued) HistomorphologiOcbservations Dose Group: Animal Number: Sex: Death Type: I I I I I I I I I I 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 F F F F F F F F F F SS SS SS SS SS SS SS SS SS ss SPINAL CORD, CERVICAL: ...... SPINALCORD, LUMBAR: ..... SPINAL CORD, THORACIC: SPLEEN: STOMACH: -dilatation,mucosal glands -edema/inflammations,ubmucosa,glandular area THYMUS: -atrophy THYROID: -ultimobranchialbody/cyst TRACHEA: ...... ...... 1 - ..... P ...... ..... - 2 P - 3 - P P - - 3 - - P URINARY BLADDER: -inflammation,chronic, focal UTERUS: -macrophages, pigmented VAGINA: - * - - - 1 3 2 3 1 3 3 3 3 3 2 ...... I-I0 418-028:PAGE J-28 ORAL {6AVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY SCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 TABLE I-7 HistomorphologicObservations Dose Group: Animal Number: Sex: Death Type: ADRENAL GLANDS: -degeneration,cystic, cortex,multifocal -hypertrophy/vacuolationc,ortex, multifocal -necrosis,cortex, focal V V V V V V V V V V 19001 1900B 19011 19020 19022 19025 19027 19028 19030 19031 F F F F F F F F F F SS SS SS SS SS SS SS SS SS SS - - <2> - - - - - - 2 - - - 2 .... BONE MARROW (STERNUM): BRAIN: ..... ..... CECUM: - <-> - - - COLON: - - <-> - - - DUODENUM: HEART: ..... <-> .... ILEUM: - <-> - - - JEJUNUM: - <-> .... KIDNEYS: -mineralization,multifocal - -nephritis,chronic, focal - LIVER: -inflammation,chronic, focal/multifocal - -necrosis,focal - - - 1 - - 1 - - 1 - I - - - - - - - - 1 LUNG_ -inflammation,interstitial,focal -macrophages,alveoli, focal I ..... i ....... LYMPH NODE, MEDIASTINAL: ....... LYMPH NODE, SUBMANDIBULAR: -hyperplasia, lymphocytic/plasmacytic MAMMARGYLAND: -necrosis, focal 2 - - - Z 2 1 2 3 * 2 - - - NERVE,SCIATIC: OVARIES: PARATHYROID: PEYER'SPATCH: RECTUM: ..... * ...... ..... ...... - <-> .... 1-11 418-028:PAGE J-29 ORAL (6AVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 TABLE I-7 (Continued) HistomorphologicObservations Dose Group: Animal Number: Sex: Death Type: V V V V V V V V V V 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 F F F F F F F F F F SS SS SS SS SS SS SS SS SS SS SKIN (GROSSLESION): -dermatitis,chronic, focal <1> SPINAL CORD, CERVICAL: ...... SPINAL CORD, LUMBAR: ..... SPINAL CORD, THORACIC: ...... SPLEEN: -atrophy - - <2> - - - STOMACH: -dilatation,mucosal glands - - - 1 - -edema/inflammations,ubmucosa,forestemach - - <2> .... -edema/inflammations,ubmucosa,glandular area - - <2> - 2 2 - - -erosion(s).glandularmucosa - - <3> .... THYMUS: -atrophy - - <4> .... THYROID: -ultimobranchialbody/cyst - P - P P P - P - TRACHEA: -inflammation,chronic,focal 1 ...... URINARY BLADDER: ..... UTERUS: -distention,lumen -macrophages,pigmented VABINA: 2 ..... 3 3 2 3 4 2 3 - 3 3 ...... 1-12 418-028:PAGE J-30 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT APPENDIX II INDIVIDUAL ANIMAL GROSS AND HISTOMORPHOLOGY DATA 418-028:PAGE J-31 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITY STUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOL NUMBER418-078 SPONSOR'SSTUDY NUMBER T-770B.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19109 DOSE GROUP: SEX: M DEATH TYPE: .............................................................................................................................................................................................................. I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLDGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LYMPH NODE, MEDIASTINAL: mastocytosis(minimal) LYMPH NODE, SUBMANDIBULAR: PROSTATE: hyperplasia,lymphocytic/plasmacyti(cmoderate) inflammation,chronic, focal/multifocal(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium (mild) ....................................................................................................... THE FOLLDWINGTISSUE(S)WERE _ITHIN NORMAL LIMITS: ADRENALGLANDS COAGULATINGGLAND HEART BONE MARROW (STERNUM) COLON ILEUM BRAIN DUODENUM JEJUNUM LIVER PEYER'S PATCH SPINAL CORD, LUMBAR TESTIS (H&E) URINARYBLADDER LUNG RECTUM SPINAL CORD, THORACIC TESTIS (PAS) NERVE, SCIATIC SEMINALVESICLES SPLEEN THYMUS ....................................................................................................... End of Record- 19109 CECUM EPIDIDYMIDES KIDNEYS PARATHYROID SPINAL CORD, CERVICAL STOMACH TRACHEA II-1 418-028:PAGE J-32 | ORAL (GAVAGE)COMBINED REPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19115 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP; DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: ADRENAL 5LANDS: vacuolatlon,cortex,diffuse (minimal) EPIDIDYMIDES: infiltration,mononuclear-cell,focal (minimal) HEART: inflammation,chronic,focal (minimal) LIVER: inflammation,chronic, focal/multifocal(minimal) THYROID: ultimobranchialbody/cyst TRACHEA: inflammation,chronic,focal (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIM NORMAL LIMITS: BONE MARROW (STERNUM) COLON BRAIN DUODENUM CECUM ILEUM KIDNEYS LUNG LYMPH NODE, MEDIASTINAL NERVE, SCIATIC PARATHYROID PEYER'SPATCH RECTUM SEMINALVESICLES SPINAL CORD, CERVICAL SPINAL CORD, THORACIC SPLEEN STOMACH TESTIS (PAS) THYMUS URINARYBLADDER ....................................................................................................... End of Record- 19115 COAGULATINGGLAND JEJUNUM LYMPH NODE, SUBMANDIBULAR PROSTATE SPINAL CORD, LUMBAR TESTIS (H&E) II-2 418-028:PAGE J-33 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBERT-7706.1 ANIMAL NUMBER: 19116 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION{S): HISTOMORPHOLOGICOBSERVATION(S): KIDNEYS:Mottledtan and red, bilateral. No microscopicchange to correlate ....................................................................................................... HISTOMORPHOLOGIOCBSERVATIONS: KIDNEYS: No microscopicchange to correlate LIVER: inflammation,chronic,focal/multfiocal (minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cmild} ....................................................................................................... THE FOLLOWINBTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGGLAND HEART LUN6 PEYER'S PATCH SPINAL CORD, CERVICAL STOMACH THYROID BONE MARROW (STERNUM) COLON ILEUM LYMPH NODE, MEDIASTINAL PROSTATE SPINAL CORD, LUMBAR TESTIS (H&E) TRACHEA BRAIN DUODENUM JEJUNUM NERVE, SCIATIC RECTUM SPINAL CORD, THORACIC TESTIS (PAS) URINARYBLADDER End of Record- 1911B CECUM EPIDIDYMIDES KIDNEYS PARATHYROID SEMINAL VESICLES SPLEEN THYMUS II-3 418-028:PAGE J-34 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19119 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): MISTOMORPHOLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTDMORPHOLOGICOBSERVATIONS: ADRENALGLANDS: vacuolation,cortex,diffuse (minimal) CECUM: inflammation,mucosa, chronic (moderate) COLON: inflammation,mucosa, chronic (mild) KIDNEYS: infiltration,mononuclear-cell,focal/multifocal(minimal) LIVER: inflammation,chronic,focal/multifocal(mild) RECTUM: inflammation,mucosa, chronic (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) BRAIN COA6ULATINGGLAND DUODENUM EPIDIDYMIDES HEART ILEUM JEJUNUM LUNG PARATHYROID LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULARNERVE, SCIATIC PEYER'S PATCH PROSTATE SEMINALVESICLES SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINALCORD, THORACIC SPLEEN STOMACH TESTIS (H&E) TESTIS (PAS) THYROID TRACHEA URINARYBLADDER ....................................................................................................... THYMUS End of Record- 19119 II-4 418-028:PAGE J-35 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENIN6TEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19122 DOSE 6ROUP: SEX: M DEATH TYPE: ====================================================================================================== I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICDBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGIOCBSERVATIONS: EPIDIDYMIDES: infiltration,mononucleer-cell,focal (minimal) LIVER: inflammation,chronic, focal/multifoca](minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytlc/plasmacyti(cminimal) TRACHEA: inflammation,chronic,focal (minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGGLAND BONE MARROW (STERNUM) COLON BRAIN DUODENUM ILEUM JEJUNUM KIDNEYS NERVE, SCIATIC PARATHYROID PEYER'S PATCH RECTUM SEMINALVESICLES SPINAL CORD, CERVICAL SPINALCORD, THORACIC SPLEEN STOMACH TESTIS (PAS) THYMUS THYROID ....................................................................................................... CECUM HEART LUNB PROSTATE SPINAL CORD, LUMBAR TESTIS (H&E) URINARY BLADDER TISSUE(S)NOT AVAILABLEFOR EVALUATION: LYMPH NODE, MEDIASTINAL ....................................................................................................... End of Record- 19122 II-5 418-028:PAGE J-36 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER:19125 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLDGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMDRPHOLOGICOBSERVATIONS: CECUM: inflammation,mucosa,chronic (moderate} LIVER: inflammation,chronic, focal/multifocal(minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cmoderate) PROSTATE: atrophy, focal/multifocal(minimal) THYROID: ultimobranchialbody/cyst ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN EPIDIDYMIDES ILEUM JEJUNUM KIDNEYS LYMPH NODE, MEDIASTINAL NERVE, SCIATIC PARATHYROID RECTUM SEMINALVESICLES SPINAL CORD, CERVICAL SPINAL CORD, THORACIC SPLEEN STOMACH TESTIS (PAS) THYMUS TRACHEA ....................................................................................................... End of Record- 19125 COAGULATINGGLAND HEART LUNG PEYER'S PATCH SPINAL CORD, LUMBAR TESTIS (H&E) URINARYBLADDER II-6 418-028:PAGE J-37 Ii ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCDLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 ANIMAL NUMBER: 19131 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLO61COBSERVATIONS: ADRENALGLANDS: vacuolation,cortex, diffuse (mild) EPIDIDYMIDES: infiltration,nmnonuclear-cell,focal (minimal) HEART: inflammation,chronic,focal (mild) LIVER: inflammation,chronic,focal/multifocal(mild) LYMPH NODE, SUBMANDIBULAR: PROSTATE: hyperplasia,lymphocytic/plasmacyti(cmild) inflammation,chronic,focal/multfiocal (minimal) TRACHEA: inflammation,chronic,focal (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: BONE MARRDW (STERNUM) BRAIN CECUM COLON DUODENUM ILEUM KIDNEYS PARATHYROID LUNG PEYER'S PATCH LYMPH NODE, MEDIASTINAL RECTUM SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINALCORD, THORACIC STOMACH THYROID TESTIS (H&E} URINARY BLADDER TESTIS (PAS) ....................................................................................................... End of Record- 19131 COAGULATINGGLAND JEJUNUM NERVE, SCIATIC SEMINALVESICLES SPLEEN THYMUS II-7 418-028:PAGE J-38 ORAL {RAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770G WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOL NUMBER418-028 SPONSOR'SSTUDY NUMBERT-770B.1 ANIMAL NUMBER: 19132 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S:} GENERAL;No gross changes. HISTOMORPHOLOGICOBSERVATION(S:) Not applicable HISTOMORPHOLOGICOBSERVATIO.N.S: LIVER: inflammation,chronic,focal/multfiocal (mild) STOMACH: edema/inflammations,ubmucosa,glandulararea (moderate) THYROID: ultimobranchail body/cyst ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINf&LAND HEART LUNG PEYER'S PATCH SPINAL CORD, CERVICAL TESTIS (H&E) URINARY BLADDER BONE MARROW (STERNUM) COLON BRAIN DUODENUM ILEUM JEJUNUM LYMPH NODE, SUBMANOIBULARNERVE, SCIATIC PROSTATE RECTUM SPINAL CORD, LUMBAR SPINAL CORD, THORACIC TESTIS (PAS) THYMUS CECUM EPIDIDYMIDES KIDNEYS PARATHYROID SEMINALVESICLES SPLEEN TRACHEA TISSUE.(SN}OT AVAILABLEFOR EVALUATION: LYMPH NODE, MEDIASTINAL ....................................................................................................... End of Record- 19132 II-8 418-028:PAGE J-39 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistemorphologyData ANIMAL NUMBER: 19134 DOSE GROUP: SEX: M DEATH TYPE: ======================================================================================================= I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL_No gross changes. Not applicable ..................... --................................................................................. HISTOMORPHOLOGICOBSERVATIONS: EPIDIDYMIDES: infiltration,mononuclear-cell,focal (minimal) KIDNEYS: cyst(s),medulla STOMACH: dilatation,mucosalglands (minimal) ede_/inflamation, submucosa,glandulararea (minimal) edema/inflammations,ubmucosa,forestomach(mild) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild) TRACHEA: inflammation,chronic,focal (minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COAGULATINGGLAND ILEUM BONE MARROW (STERNUM) COLON JEJUNUM BRAIN DUODENUM LIVER LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULARNERVE, SCIATIC PEYER'SPATCH PROSTATE RECTUM SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINAL CORD, THORACIC TESTIS (H&E) TESTIS (PAS) THYMUS ....................................................................................................... End of Record- 19134 CECUM HEART LUNG PAR.ATHYROID SEMINALVESICLES SPLEEN URINARYBLADDER II-9 418-028:PAGE J-40 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER:19135 DOSE GROUP: SEX: M DEATH TYPE: ======================================================================================================= I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOL061OCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: KIDNEYS: infiltration,mononuclear-cell,focal/multlfocal(minimal} LIVER: inflammation,chronic, focal/multifocal(minimal) PROSTATE: inflammation,chronic, focal/multlfocal(minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGGLAND BONE MARROW (STERNUM) COLON BRAIN DUODENUM HEART ILEUM JEJUNUM LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULARNERVE, SCIATIC PEYER'SPATCH RECTUM SEMINALVESICLES SPINALCORD, LUMBAR SPINAL CORD, THORACIC SPLEEN TESTIS (H&E) TRACHEA TESTIS (PAS) URINARYBLADDER THYMUS ....................................................................................................... End of Record- 19135 CECUM EPIDIDYMIDES LUNG PARATHYROID SPINALCORD, CERVICAL STOMACH THYROID 11-10 418-028:PAGE J-41 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY$CREENIN6TEST PROTOCOL NUMBER418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19102 SEX: M Appendix II IndividualAnimal 6ross and HistomorphologyData DOSE GROUP: DEATH TYPE: II Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMDRPHDLDGICOBSERVATION(S): KIDNEYS: Bilateral-mottled. No microscopicchange to correlate ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: KIDNEYS: No microscopicchange to correlate LIVER: inflammation,chronic, focal/multifocal(mild) THYROID: ultimobranchialbody/cyst ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: KIDNEYS ....................................................................................................... End of Record- 19102 II-11 418-02_:PAGE 1-42 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-77OB _ITH THE REPRODUCTION/DEVELOPMENTATLOXICITY$CREENIHBTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.I ANIMAL NUMBER: 19106 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP.: II DEATH TYPE: Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHDLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic,focal/multifocal(mild) THYROID: ultimobranchialbody/cyst ....................................................................................................... End of Record- 19106 II-12 418-02_'.PAGE3-43 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENIN6TEST PROTOCOL NUMBER418-0Z8 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19108 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: II Sacrifice-Scheduled GROSS OBSERVATIONI_): GENERAL:No gross changes. HISTOMORPHOLOGIOCBSERVATION(S): Not applicable HISTOMORPHOLOGICOBSERVATIONS: LIVER: lipidosis,tension, focal inflammation,chronic, focal/multifocal(minimal) THE FOLLOWINGTISSUE(S) WERE WITHIN NORMAL _[MITS: THYROID ....................................................................................................... End of Record- 19108 II-13 418-028:PAGE J-44 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 41B-O2B SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER:19110 SEX: M =====================_================================================================================= DDSE GROUP: DEATH TYPE: II Sacrifice-Scheduled 6ROSS OBSERVATION(S): GENERAL:No gross changes. HISTOMORPHOLOGICOBSERVATION(S}: Not applicable HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocaI(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild) ....................................................................................................... End of Record- 19110 II-14 418-028:PAGE J-45 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY SCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER:19113 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: II Sacrifice-Schedueld GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): KIDNEYS:Left- pelvis, slight dilation, dilatation,pelvis ....................................................................................................... HISTOMORPHOLOBICOBSERVATIONS: KIDNEYS: dilatation,pelvis (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: LIVER THYROID ....................................................................................................... End of Record- 19113 11-15 418-028:PAGE J-46 ORAL (GAVAGE)COMBINEDREPEATED DOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PRDTOCDLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 ANIMAL NUMBER: 19120 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: II Sacrifice-Scheduled GROSS OBSERVATION(S}: HISTOMORPHOLOGICOBSERVATION(S): GENERAL: No gross _hanges. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic,focal/multifocal{minimal) lipidosis,tenBion, focal ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record-19120 II-16 418-028:PAGE J--47 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY5CREENIN6TEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.I ANIMAL NUMBER: 19129 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE 6ROUP.: IT DEATH TYPE: Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOL061COBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifoeal(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium (minimal) ....................................................................................................... End of Record- 19129 II-1/ 418-028:PAGE J-48 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY5CREENIN6TEST PROTOCOL NUMBER41B-OZB SPONSOR'SSTUDY NUMBER T-7706.I Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19136 SEX: M ======================_===_=========_==_=========_=====_=_m=s==_=_===s============================ DosE GROUP: DEATH TYPE: II Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): KIDNEYS: Bilateral-mottled, dilatation,pelvis ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: KIDNEYS: dilatation,pelvis (minimal) LIVER: vacuolation,hepatocellular,multifocal (minimal) THYROID: ultimobranchialbody/cyst ....................................................................................................... End of Record-19136 II-18 418-028:PAGE J-49 ORAL (GAVAGE)COMBINED REPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19138 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: II Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ...................... --................................................................................ HISTOMORBHOLOGICOBSERVATIONS: THYROID: ultimobranchialbody/cyst ....................................................................................................... THE FOLLOWINGTISSUE(S}WERE WITHIN NORMAL LIMITS: LIVER ....................................................................................................... End of Record- 19138 II-19 418-028:PAGE J-50 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENIN6TEST PROTOCOLNUMBER 418-02B SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19139 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: II Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOBICOBSERVATIONS: THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: LIVER End of Record- 19139 II-20 418-028"PAGEJ-51 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITY STUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTTAOLXICITY SCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 ANIMAL NUMBER: 19101 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHDLDGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: LIVER THYROID ....................................................................................................... End of Record- 19101 II-Zl 418-028:PAGE J-52 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19105 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP; DEATH TYPE: Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: _nflammation,chronic, focal/multifocalIminimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record- 19105 11-22 418-028:PAGE J-53 ORAL (6AVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19107 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSEGROUP: DEATH TYPE: Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocal(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(minimal) ultimobranchialbody/cyst ....................................................................................................... End of Record- 19107 11-23 418-028:PAGE J-54 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19112 DOSE GROUP: SEX: M DEATH TYPE: ======================================================================================================= Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTDMORPHOLOGICOBSERVATION{S): GENERAL:No gross changes. Not applicable ....................... _ ............................................................................... MISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocal(minimal) THYROID: ultimobranchialbody/cyst ....................................................................................................... End of Record- 19112 11-24 418-028:PAGE J-55 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTTAOLXICITY SCREENINGTEST PROTOCOLNUMBER 418-02B SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19114 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE BROUP: DEATH TYPE: Ill Sacrifice-Scheduled GROSS OBSERVATION(S}: HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multlfocal{minimal) THYROID: ultimobranchialbody/cyst ....................................................................................................... End of Record- 19114 11-25 418-028:PAGE J-56 ORAL {GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINBTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.I Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19121 SEX: M DOSE 6ROUP: DEATH TYPE: ======================================================================================================= Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: THYROID: End of Record- 19121 inflammation,chronic, focal/multifocal(minimal) vacuolation,hepatoce]lular,multifocal (mild) ultimobranchialbody/cyst 11-26 418-028:PAGE J-57 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENIN6TEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19123 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION_S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocal(minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record- 19123 II-2/ 418-028:PAGE J-58 ORAL (GAVAGE}COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770G WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 41B-D2B SPONSOR'SSTUDY NUMBER T-770B.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19130 SEX: M DOSEGROUP: DEATH TYPE: ======================================================================================================= Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION{S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic,focal/multfiocal (minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild} ....................................................................................................... End of Record- 19130 II-28 418-028:PAGE J-59 ORAL (GAVAGE)COMBINED REPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19137 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION(S): GENERAL: No gross changes. Not applicable ........................ --.............................................................................. THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: LIVER THYROID ....................................................................................................... End of Record- 19137 II-29 418-028:PAGE J-60 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 41B-028 SPONSOR'SSTUDY NUMBER T-7706.I Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER:19146 DOSE GROUP: SEX: M DEATH TYPE: ====================================================================================================== Ill Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: vacuolation,hepatocellularm,idzonal (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record-19146 11-30 418-028:PAGE J-61 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTESI PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19100 DOSE GROUP: SEX: M DEATH TYPE: ======================================================================================================= IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION{S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: hypertrophy,hepatocellular,centrilobular{minimal) inflammation,chronic,focal/multifocal(minimal) THYROID: h_o_ertrophy/hyperplasifao,llicularepithelium(minimal) ....................................................................................................... End of Record-19100 II-31 418-028:PAGE J-62 | ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19103 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP.: IV DEATH TYPE: Sacrifice-Scheduled GROSS OBSERVATION(S): HISTDMORPHOLOGIOCBSERVATION(S}: GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: hypertrophy,hepatoeellular,eentrilobular(minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record- 19103 II-32 418-028:PAGE J-63 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENIN6TEST PROTOCOLNUMBER 418-02B SPONSOR'SSTUDY NUMBER T-7706.I ANIMAL NUMBER: 19104 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocal(minimal) hypertrophy,hepatoceIlular,centrilobular(mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record- 19104 II-33 418-028:PAGE J-64 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19118 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocal(minimal) hypertrophy,hepatocellular,centrilobular(minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record-19118 II-34 418-028:PAGE J-65 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19133 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: hypertrophy,hepatocellular,centrilobular(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium (minimal) ultimobranchialbody/cyst ....................................................................................................... End of Record- 19133 11-35 418-028:PAGE J-66 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PRDTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19141 DOSE GROUP: SEX: M DEATH TYPE: ======================================================================================================= IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGIOCBSERVATIONS: LIVER: hypertrophy,hepatocellular,centrilobular(minimal) THYROID: ultimobranchialbody/cyst ....................................................................................................... End of Record- 19141 11-36 418-028:PAGE J-67 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOL NUMBER418-028 SPONSOR'SSTUDY NUMBERT-7706.1 ANIMAL NUMBER: 19142 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPMOLOGIOCBSERVATION(S): EPIDIDYMIDES:Bilateral-small; cauda epididymis, hypospermia left, small, TESTES: Bilateral-small, exfoliatedspermatogeniccells degeneration,multifocal,TESTIS (PAS) degeneration,multifocal,TESTIS (H&E) ....................................................................................................... MISTOMORPHOLOBICOBSERVATIONS: EPIDIDYMIDES: hypospermia(marked) exfoliatedspermatogeniccells (moderate) LIVER: inflammation,chronic, focal/multifocal(minimal) hypertrophy,hepatocellular,centrilobular(minimal) TESTIS (H&E): degeneration,multlfocal(mild) TESTIS (PAS): degeneration,multifoeal(mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record-19142 II-37 418-028:PAGE J-68 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER:19144 SEX: M Appendix II IndividualAnimal 6ross and HistomorphologyData DOSE GROUP: DEATH TYPE: IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic,focal/multifocal(minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S}WERE WITHIN NORMAL LIMITS: THYROID ....................................................................................................... End of Record- 19144 II-38 418-028:PAGE J-69 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19148 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: THYROID: End of Record-19148 inflammation,chronic, focal/multifooal(minimal) hypertrophy,hepatocellular,centrilobular(minimal) hypertrophy/hyperplasiaf,ollicularepithelium (mild) II-39 418-028:PAGE J-70 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 41B-028 SPONSOR'SSTUDY NUMBERT-?7OB.1 ANIMAL NUMBER:19150 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: IV Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERALzNo gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: hypertrophy,hepatocellular,centrilobular(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild) ....................................................................................................... End of Record- 19150 II-40 418-028:PAGE J-71 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19111 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATIDN(SI: GASTROINTESTINALTRACT: Jejunum-diverticulum, l.SxO.BxO.4cm. diverticulum,JEJUNUM ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: JEJUNUM: diverticulum KIDNEYS: basophilia/degenerationc,ortical tubules, focal/multifocal (minimal) cyst(s),medulla infiltration,mononuclear-cell,focal/multifocal(minimal) LIVER: hypertrophy,hepatocellular,centrilobular(mild) inflammation,chronic,focal/multfiocal (minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cminimal) PEYER'SPATCH: mineralization,focal (minimal) PROSTATE: inflammation,chronic,focal/multfiocal (minimal) THYROID: ultimobranchialbody/cyst hypertrophy/hyperplasiaf,ollicularepithelium(moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAl LIMITS: ADRENAL GLANDS COAGULATINGGLAND BONE MARROW (STERNUM) COLON BRAIN DUODENUM HEART NERVE, SCIATIC SPINAL CORD, CERVICAL STOMACH TRACHEA ILEUM PARATHYROID SPINAL CORD, LUMBAR TESTIS (H&E) URINARY BLADDER LUNG RECTUM SPINAL CORD, THORACIC TESTIS (PAS) ....................................................................................................... End of Record- 19111 CECUM EPIDIDYMIDES LYMPH NODE, MEDIASTINAL SEMINALVESICLES SPLEEN THYMUS 11-41 I 418-028:PAGE J-72 D ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19117 DOSE GROUP: SEX: M DEATH TYPEi ==_=_================================================================================================ V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: hypertrophy,hepatocellular,centrilobular(minimal) LYMPH NODE, MEDIASTINAL: mastocytosis(minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cmoderate) STOMACH: edema/inflammations,ubmucosa,forestomach(mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COAGULATINGGLAND HEART LUNG BONE MARROW (STERNUM) COLON ILEUM NERVE, SCIATIC BRAIN DUODENUM JEJUNUM PARATHYROID PROSTATE SPINAL CORD, LUMBAR TESTIS (PAS) URINARYBLADDER RECTUM SPINAL CORD, THORACIC THYMUS SEMINALVESICLES SPLEEN THYROID ....................................................................................................... End of Record- 19117 CECUM EPIDIDYMIDES KIDNEYS PEYER'SPATCH SPINAL CORD, CERVICAL TESTIS (H&E) TRACHEA II-42 418-028:PAGE J-73 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOL NUMBER418-028 SPONSOR'SSTUDY NUMBER T-770B.I ANIMAL NUMBER: 19124 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS; EPIDIDYMIDES: infiltration,mononuclear-oell,focal (minimal) KIDNEYS: nephritis,chronic,focal (minimal) cyst(s), medulla LIVER: hypertrophy,hepatoeellular,centrilobular(mild) LYMPH NODE, MEDIASTINAL: mastocytosis(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGGLAND BONE MARROW (STERNUM) COLON BRAIN DUODENUM ILEUM NERVE, SCIATIC JEJUNUM PARATHYROID LUNG PEYER'SPATCH RECTUM SPINAL CORD, THORACIC TESTIS (PAS) SEMINALVESICLES SPLEEN THYMUS SPINAL CORD, CERVICAL STOMACH TRACHEA ....................................................................................................... End of Record- 19124 CECUM HEART LYMPH NODE, SUBMANDIBULAR PROSTATE SPINAL CORD, LUMBAR TESTIS (H&E) URINARY BLADDER II-43 418-028:PAGE J-74 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19126 DOSE GROUP: SEX: M DEATH TYPE: ========================_=======================================================_====================== V Sacrifice-Scheduled DROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................... _ ............................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocal(minimal) hypertrophy,hepatocellular,centrilobular(mild) PROSTATE: inflammation,chronic, focal/multifocal(minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGBLAND HEART LUNG PARATHYROID SPINALCORD, CERVICAL STOMACH THYROID BONE MARROW (STERNUM) COLON ILEUM LYMPH NODE, MEDIASTINAL PEYER'SPATCH SPINAL CORD, LUMBAR TESTIS (H&E) TRACHEA BRAIN DUODENUM CECUM EPIDIDYMIDES JEJUNUM KIDNEYS LYMPH NODE, SUBMANDIBULARNERVE, SCIATIC RECTUM SEMINALVESICLES SPINAL CORD, THORACIC SPLEEN TESTIS (PAS) URINARYBLADDER THYMUS End of Record- 19126 11-44 418-028:PAGE J-75 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-77OB WITM THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19127 SEX: M DOSE GROUP: DEATH TYPE: ====================================================================================================== V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION{S}: GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: ADRENALGLANDS: vacuolation,cortex, diffuse (mild) LIVER: hypertrophy,hepatocellular,centrilobular(moderate) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cminimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) COLON BRAIN DUODENUM CECUM EPIDIDYMIDES ILEUM JEJUNUM KIDNEYS LYMPH NODE, MEDIASTINAL NERVE, SCIATIC PARATHYROID PROSTATE SPINAL CORD, LUMBAR RECTUM SPINAL CORD, THORACIC SEMINALVESICLES SPLEEN TESTIS (H&E) URINARY BLADDER TESTIS (PAS) THYMUS ....................................................................................................... End of Record- 19127 COAGULATINGGLAND HEART LUNG PEYER'S PATCH SPINAL CORD, CERVICAL STOMACH TRACHEA 11-45 418-028:PAGE J-76 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY5GREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.I Appendix II IndividualAnimal Gross and HistumorphologyData ANIMAL NUMBER: 19128 DOSE GROUP: SEX: M DEATH TYPE: ======================================================================================================= V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S1: GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: CECUM: inflammation,mucosa, chronic (mild) LIVER: inflammation,chronic, focal/multifocal(minimal) hypertrophy,hepatocellular,centrilobular(mild) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cmoderate) PROSTATE: atrophy, focal/multifocal(moderate) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAl LIMITS: ADRENALGLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN EPIDIDYMIDES ILEUM JEJUNUM KIDNEYS LYMPH NODE, MEDIASTINAL NERVE, SCIATIC PARATHYROID RECTUM SEMINAL VESICLES SPINAL CORD, CERVICAL SPINAL CORD, THORACIC SPLEEN STDMACH TESTIS {PAS) THYMUS TRACHEA ....................................................................................................... End of Record- 19128 COAGULATINGGLAND HEART LUNG PEYER'SPATCH SPINALCORD, LUMBAR TESTIS (H&E) URINARY BLADDER 11-46 418-028:PAGE J-77 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-02B SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19140 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL_No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: inflammation,chronic, focal/multifocal(minimal) hypertrophy,hepatocellular,centrilobular(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGGLAND HEART LUNG PARATHYROID BONE MARROW (STERNUM) COLON ILEUM LYMPH NDDE, MEDIASTINAL PEYER'SPATCH BRAIN CECUM DUODENUM EPIDIDYMIDES JEJUNUM KIDNEYS LYMPH NODE, SUBMANDIBULARNERVE, SCIATIC PROSTATE RECTUM SEMINALVESICLES SPLEEN THYMUS SPINAL CORD, CERVICAL STOMACH TRACHEA SPINAL CORD, LUMBAR TESTIS (H&E) URINARYBLADDER SPINALCORD, THORACIC TESTIS (PAS) ....................................................................................................... End of Record- 19140 11-47 418-028:PAGE J-78 ORAL (GAVAGE)COMBINED REPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.I ANIMAL NUMBER: 19145 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: KIDNEYS: infiltration,mononuclear-cell,focal/multifocal(minimal) LIVER: hypertrophy,hepatocellular,centrilobular(mild) LYMPH NODE, SUBMANDIBULAR: RECTUM: hyperplasia,lymphocytlc/plasmacyti(cmoderate) inflammation,mucosa, chronic (minimal) TRACHEA: inflammation,chronic,focal (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COAGULATINGGLAND HEART BONE MARROW (STERNUM) COLON ILEUM BRAIN DUODENUM JEJUNUM LYMPH NODE, MEDIASTINAL PROSTATE SPINAL CORD, THORACIC TESTIS {PAS) NERVE, SCIATIC SEMINALVESICLES SPLEEN THYMUS PARATHYROID SPINALCORD, CERVICAL STOMACH THYROID ....................................................................................................... End of Record- 19145 CECUM EPIDIDYMIDES LUNG PEYER'S PATCH SPINAL CORD, LUMBAR TESTIS (H&E) URINARY BLADDER 11-48 418-028:PAGE J-79 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19149 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: hypertrophy,hepatocellular,centrilobular{minimal) THYROID: ultimobranchialbody/cyst hypertrophy/hyperplasiaf,ollicularepithelium(moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)W_RE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGGLAND HEART LUNG PARATHYROID BONE MARROW (STERNUM) COLON ILEUM LYMPH NODE, MEDIASTINAL PEYER'SPATCH BRAIN CECUM DUODENUM EPIDIDYMIDES JEJUNUM KIDNEYS LYMPH NODE, SUBMANDIBULARNERVE, SCIATIC PROSTATE RECTUM SEMINAL VESICLES SPLEEN THYMUS SPINALCORD, CERVICAL STOMACH TRACHEA SPINAL CORD, LUMBAR TESTIS (H&E) URINARYBLADDER SPINAL CORD, THORACIC TESTIS (PAS) ....................................................................................................... End of Record-19149 II-49 418-028:PAGE J-80 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19152 SEX: M Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOL061COBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGIOCBSERVATIONS: HEART: inflammation,chronic,focal (minimal) LIVER: inflammation,chronic, foeal/multifocal(minimal) hypertrophy,hepatocellular,centrilobular(minimal) LYMPH NODE, SUBMANDIBULAR: PROSTATE: hyperplasia,lymphocytio/plasmaeyti(cmoderate) inflammation,chronic, focal/multifocal(minimal) atrophy, focal/multifocal(minimal) THYROID: hypertrophy/hyperplasiaf,ollicularepithelium(mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COAGULATINGGLAND ILEUM BONE MARROW (STERNUM) COLON JEJUNUM BRAIN DUODENUM KIDNEYS LYMPH NODE, MEDIASTINAL RECTUM SPINAL CORD, THORACIC TESTIS (PAS) NERVE, SCIATIC SEMINALVESICLES SPLEEN THYMUS PARATHYROID SPINAL CORD, CERVICAL STOMACH TRACHEA ....................................................................................................... End of Record- 19152 CECUM EPIDIDYMIDES LUNG PEYER'SPATCH SPINAL CORD, LUMBAR TESTIS (H&E) URINARY BLADDER 11-50 418-028:PAGE J-81 ORAL (GAVAGE}COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY5GREENINGTESI PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7708.I ANIMAL NUMBER: 19012 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): GENERAL:No gross changes. ...................................................................... HISTOMORPHOLOGIOCBSERVATION(S): Not applicable ................................. HISTOMORPHOLOGICOBSERVATIONS: STOMACH: dilatation,mucosalglands (minimal) THYROID: ultimobranohialbody/cyst UTERUS: macrophages,pigmented(moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS BDNE MARROW (STERNUM) BRAIN COLON JEJUNUM DUODENUM KIDNEYS HEART LIVER LYMPH NODE, MEDIASTINAL LYMPH NODE, SUBMANDIBULARMAMMARYGLAND OVARIES PARATHYROID PEYER'S PATCH SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPINAL CORD, THORACIC THYMUS TRACHEA URINARYBLADDER ....................................................................................................... End of Record- 19012 CECUM ILEUM LUNG NERVE, SCIATIC RECTUM SPLEEN VAGINA II-51 418-028:PAGE J-82 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTIDN/DEVELOPMENTATLOXICITYSCREENINGTEST PRDTOCDLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19019 SEX: F DOSE GROUP_ DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION{S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cmild) UTERUS: macrophages,pigmented (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN HEART JEJUNUM KIDNEYS LIVER LYMPH NODE, MEDIASTINAL MAMMARYGLAND NERVE, SCIATIC PARATHYROID SPINAL CORD, LUMBAR PEYER'SPATCH SPINAL CORD, THORACIC RECTUM SPLEEN THYMUS VAGINA THYROID TRACHEA ....................................................................................................... End of Record- 19019 CECUM ILEUM LUNG OVARIES SPINAL CORD, CERVICAL STOMACH URINARY BLADDER II-52 418-028:PAGE J-83 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19021 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData DOSE 6ROUP: DEATH TYPE: I Sacrifice-Scheduled 6ROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): HEAD: Head- scab, O.2cmxO.4cm. dermatitis,chronic,focal, SKIN (GROSS LESION) ....................................................................................................... HISTOMORPHOL061COBSERVATIONS: LIVER: inflammation,chronic,focal/multifocal(minimal) LYMPH NODE, MEDIASTINAL: congestion (minimal) SKIN (GROSS LESION): THYROID: dermatitis,chronic,focal (mild) ultimobranchialbody/cyst UTERUS: macrophages,pigmented (moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN HEART JEJUNUM KIDNEYS LUNG MAMMARY GLAND NERVE, SCIATIC OVARIES PEYER'SPATCH RECTUM SPINAL CORD, CERVICAL SPINAL CORD, THDRAClC TRACHEA SPLEEN VAGINA STOMACH ....................................................................................................... CECUM ILEUM LYMPH NODE, SUBMANDIBULAR PARATHYROID SPINAL CORD, LUMBAR THYMUS TISSUE(S) NOT AVAILABLEFOR EVALUATION: URINARYBLADDER .......................... End of Record- 19021 --............................................................................. Il-S3 418-028:PAGE J-84 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 416-028 SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19023 DOSE GROUP: SEX: F DEATH TYPE: ======================================================================================================= I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION{S}: GENERALzNo gross changes. Not applicable ....................... --............................................................................... HISTOMORPHOLOGICOBSERVATIONS: LUNG: infiltration,polymorphonuclearp,erivascular/peribronchial (mild) LYMPH NODE, MEDIASTINAL: mastocytosis(minimal) STOMACH: edema/inflammations,ubmucosa,glandulararea (mild) UTERUS: macrophages,pigmented (minimal) ....................................................................................................... THE FOLLOWINGTISSUE(S}WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN HEART JEJUNUM KIDNEYS LIVER MAMMARYGLAND NERVE, SCIATIC OVARIES PEYER'S PATCH RECTUM SPINALCORD, CERVICAL SPINAL CORD, THORACIC TRACHEA SPLEEN URINARY BLADDER THYMUS VAGINA ....................................................................................................... End of Record- 19023 CECUM ILEUM LYMPH NODE, SUBMANDIBULAR PARATHYROID SPINAL CORD, LUMBAR THYROID II-54 418-028:PAGE J-85 ! ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY5CREENII_TiEST PROTOCOLNUMBER 418-02B SPONSOR'SSTUDY NUMBER T-7706.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19041 SEX: F = = =m_=_m======_========_====_========_=========_==========_=_===-_===_============_==_========== DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S}: GENERAL:No gross changes. HISTOMORPHOLOBICOBSERVATION(S): Not applicable HISTOMORPHOLDGICOBSERVATIONS: LYMPH NDDE, SUBMANDIBULAR: STOMACH: hyperplasia,lymphoeytic/plasmacyti(cmoderate) edema/inflammations,ubmucosa,glandulararea (moderate) THYROID: ultimobranchialbody/cyst UTERUS: maorophages,pigmented (moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN HEART JEJUNUM KIDNEYS LIVER LYMPH NODE, MEDIASTINAL MAMMAR_ GLAND NERVE, SCIATIC PARATHYROID SPINAL CORD, LUMBAR PEYER'SPATCH SPINALCORD, THORACIC RECTUM SPLEEN TRACHEA URINARY BLADDER VAGINA ....................................................................................................... End of Record- 19041 CECUM ILEUM LUNG OVARIES SPINAL CORD, CERVICAL THYMUS II-55 418-028:PAGE J-86 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRDDUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19042 SEX: F Appendix 11 IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION{S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTDMORPHOLOGICOBSERVATIONS: OVARIES: cyst(s), intraovarian THYROID: ultimobranchialbody/cyst UTERUS: macrophages,pigmented (moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COLON JEJUNUM LYMPH NODE, MEDIASTINAL PARATHYROID SPINALCORD, LUMBAR THYMUS BONE MARROW {STERNUM) BRAIN DUODENUM HEART KIDNEYS LIVER LYMPH NODE, SUBMANDIBULARMAMMARYGLAND PEYER'SPATCH RECTUM SPINAL CORD, THORACIC SPLEEN TRACHEA URINARYBLAODER ....................................................................................................... End of Record- 19042 CECUM ILEUM LUN6 NERVE, SCIATIC SPINALCORD, CERVICAL STOMACH VAGINA II-56 418-028:PAGE J-87 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19044 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP:. I DEATH TYPE: Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: KIDNEYS: LYMPH NODE, MEDIASTINAL: nephritis,chronic,focal (minimal) congestion(minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocy%ic/plasmacyti(cmild) hyperplasia,lymphocy%ic/plasmacyti(cmoderate) UTERUS: macrophages,pigmented (moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS BONE MARROW (STERNUM) BRAIN COLON JEJUNUM DUODENUM LIVER HEART LUNG NERVE, SCIATIC OVARIES PARATHYROID RECTUM SPLEEN TRACHEA SPINAL CORD, CERVICAL STOMACH URINARY BLADDER SPINALCORD, LUMBAR THYMUS VAGINA ....................................................................................................... End of Record- 19044 CECUM ILEUM MAMMARYGLAND PEYER'SPATCH SPINAL CORD, THORACIC THYROID lI-B7 418-028:PAGE J-88 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITY$CREENIN6TEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19050 SEX: F Appendix IT IndividualAnimal Gross and Histo_rphology Data DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: ADRENAL GLANDS: vacuolation,zona glomerulosa(mild) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cminimal) UTERUS: macrophages,pigmented(moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) DUODENUM BRAIN HEART CECUM ILEUM KIDNEYS MAMMARY BLAND PEYER'SPATCH SPINAL CORD, THORACIC THYROID LIVER NERVE, SCIATIC RECTUM SPLEEN TRACHEA LUNG OVARIES SPINALCORD, CERVICAL STOMACH URINARYBLADDER ....................................................................................................... End of Record- 19050 COLON JEJUNUM LYMPH NODE, MEDIASTINAL PARATHYROID SPINAL CORD, LUMBAR THYMUS VAGINA 11-58 418-028:PAGE J-89 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOL NUMBER418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19053 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData DDSE GROUP: DEATH TYPE: I Sacrifice-Schedueld GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION[S}: GENERAL:No gross changes. Not applicable ........................ --.............................................................................. HISTOMORPHOLOGICOBSERVATIONS: ADRENAL GLANDS: CECUM: LYMPH NODE, SUBMANDIBULAR: THYMUS: THYROID: UTERUS: ....................................................................................................... vacuolation,zona glomerulosa(mild) inflammation,mucosa,chronic (moderate) hyperplasia,lymphooytic/plasmacyti(cmild) atrophy (moderate) ultimobranchialbody/cyst macrophages, pigmented (moderate) THE FOLLOWINGTISSUE(S) WEREWITHIN NORMALIMITS: BONE MARROW (STERNUM) HEART LIVER NERVE, SCIATIC BRAIN ILEUM LUNG OVARIES COLON JEJUNUM LYMPH NODE, MEDIASTINAL PARATHYROID RECTUM SPLEEN VAGINA SPINAL CORD, CERVICAL STOMACH SPINAL CORD, LUMBAR TRACHEA ....................................................................................................... End of Record- 19053 DUODENUM KIDNEYS MAMMARYGLAND PEYER'S PATCH SPINAL CORD, THORACIC URINARYBLADDER I1-59 418-028:PAGE J-90 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTTAOLXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.1 ANIMAL NUMBER: 19065 SEX: F Appendix II IndividualAnimal Gross and MistomorphologyData DOSE GROUP: DEATH TYPE: I Sacrifice-Scheduled GROSS OBSERVATION(S): HISTDMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: CECUM: LIVER: LYMPH NODE, SUBMANDIBULAR: URINARY BLADDER: UTERUS: inflammation,mucosa, chronic (mild) inflammation,chronic, focal/multifocal(minimal) hyperplasia,lymphocytic/plasmacyt_(cmild) inflammation,chronic,focal (minimal) macrophages,pigmented (mild) THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS DUODENUM KIDNEYS NERVE, SCIATIC RECTUM SPLEEN TRACHEA BONE MARROW (STERNUM) HEART LUNG OVARIES SPINAL CORD, CERVICAL STOMACH VAGINA BRAIN ILEUM LYMPH NODE, MEDIASTINAL PARATHYROID SPINALCORD, LUMBAR THYMUS ....................................................................................................... End of Record- 19065 COLON JEJUNUM MAMMARYGLAND PEYER'S PATCH SPINAL CORD, THORACIC THYROID II-60 418-028:PAGE J-91 ORAL (GAVAGE}COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PRDTOCOLNUMBER 41B-D2B SPONSOR'SSTUDY NUMBER T-770B.I Appendix II IndividualAnimal 6ross and HistomorphologyData ANIMAL NUMBER: 19001 SEX: F =======_=====_=_=_=======_=_==_======_=_z_==_====_=_===================================== DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S}: GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LUNG: inflammation,interstitial,focal (m_nimal) macrophages,alveoli,focal (minimal) TRACHEA: inflammation,chronic,focal (minimal) UTERUS: macrophages,plg_nted (moderate) distention,lumen (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S}WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN HEART JEJUNUM KIDNEYS LYMPH NODE, SUBMANDIBULARMAMMARYGLAND PARATHYROID PEYER'SPATCH SPINAL CORD, LUMBAR SPINAL CORD, THORACIC THYMUS THYROID LIVER NERVE, SCIATIC RECTUM SPLEEN URINARY BLADDER ....................................................................................................... End of Record- 19001 CECUM ILEUM LYMPH NODE, MEDIASTINAL OVARIES SPINALCORD, CERVICAL STOMACH VAGINA II-Bl 418-028:PAGE J-92 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770G.1 ANIMAL NUMBER:19006 SEX: F Appendix II IndividualAnimal Gross and Histo_orphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cmild) THYROID: ultimobranchialbody/cyst UTERUS: macrophages,pign_nted (mild) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENAL GLANDS COLON JEJUNUM BONE MARROW (STERNUM) DUODENUM KIDNEYS BRAIN HEART LIVER LYMPH NODE, MEDIASTINAL PARATHYROID SPINAL CORD, LUMBAR THYMUS MAMMARY6LAND PEYER'SPATCH SPINAL CDRD, THORACIC TRACHEA NERVE, SCIATIC RECTUM SPLEEN URINARYBLADDER ....................................................................................................... End of Record-1900B CECUM ILEUM LUNG OVARIES SPINAL CORD, CERVICAL STOMACH VAGINA II-62 418-028:PAGE J-93 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19011 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData Dose GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: ADRENAL GLANDS: LIVER: UTERUS: necrosis,cortex,focal (mild) Not remarkable macrophages,pigmented(moderate) THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) DUODENUM BRAIN HEART CECUM ILEUM KIDNEYS MAMMARY GLAND LUNG NERVE, SCIATIC LYMPH NODE, MEDIASTINAL OVARIES PEYER'SPATCH RECTUM SPINAL CORD, CERVICAL SPINAL CORD, THORACIC THYROID SPLEEN TRACHEA STOMACH URINARYBLADDER ....................................................................................................... End of Record- 19011 COLON JEJUNUM LYMPH NODE, SUBMANDIBULAR PARATHYROID SPINAL CORD, LUMBAR THYMUS VAGINA II-63 418-028:PAGE J-94 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOL NUMBER418-028 SPONSOR'SSTUDY NUMBERT-7706.1 ANIMAL NUMBER: 19020 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): ADRENAL6LANDS: Bilateral-large, degeneration,cystic,cortex,multifocal GASTROINTESTINALTRACT:Abdominal distention, No mlcroscopicchange to correlate,DUODENUM large and small intestinesdistendedwith gas. No mlcroscopicchange to correlate,JEJUNUM No mlcroscopicchange to correlate,ILEUM No mlcroscopicchange to correlate,COLON No m_croscopicchange to correlate,CECUM No m_croscopicchange to correlate,RECTUM SPLEEN: Small. atrophy STOMACH: Fundicmucosa surfacecontains edema/inflammations,ubmucosa,glandulararea approximately12 black areas rangingin size edema/inflammations,ubmucosa,forestomach from pinpointto O.3cm in diameter, eroslon(s),glandularmucosa THYMUS: Small. atrophy ........................................................................................................ HISTOMORPHOLOBICOBSERVATIQNS: ADRENALGLANDS: degeneration,cystic,cortex, multifocal(mild) CECUM: No microscopicchange to correlate COLON: No microscopicchange to correlate DUODENUM: No microscopicchange to correlate ILEUM: No microscopicchange to correlate JEJUNUM: LIVER: No microscopicchange to correlate necrosis,focal (minimal) LYMPH NODE, SUBMANDIBULAR: RECTUM: hyperplasia,lymphocytic/plasmacyti(cmild) No microscopicchange to correlate SPLEEN: atrophy (mild) STOMACH: erosion(s),glandularmucosa (moderate) edema/inflammations,ubmucosa,glandulararea (mild) edema/inflammation,submucosa,forestomach(mild) THYMUS: THYROID: atrophy (marked) ultimobranchialbody/cyst UTERUS: macrophages,pigmented (marked) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) DUODENUM BRAIN HEART CECUM ILEUM KIDNEYS LUNG LYMPH NODE, MEOIASTINAL NERVE, SCIATIC OVARIES PARATHYROID RECTUM TRACHEA SPINAL CORD, CERVICAL URINARY BLADDER SPINAL CORD, LUMBAR VAGINA ....................................................................................................... End of Record- 190Z0 COLON JEJUNUM MAMMARY BLAND PEYER'SPATCH SPINAL CORD, THORACIC II-B4 418-028:PAGE J-95 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER:19022 SEX: F DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGIOCBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOL061COBSERVATIONS: LIVER: inflammation,chronic,focal/_Itifocal (minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphocytic/plasmacyti(cmild) MAMMARY GLAND: necrosis,focal (minimal) THYROID: ultimobranohialbody/cyst UTERUS: macrophages,pigmented (mild) ........................................................................................................ THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS BONE MARROW (STERNUM) BRAIN COLON DUODENUM HEART JEJUNUM KIDNEYS LUNG NERVE, SCIATIC OVARIES PARATHYROID RECTUM SPINAL CORD, CERVICAL SPINALCORD, LUMBAR SPLEEN STOMACH THYMUS URINARYBLADDER VAGINA ....................................................................................................... End of Record- 19022 CECUM ILEUM LYMPH NODE, MEDIASTINAL PEYER'S PATCH SPINAL CORD, THORACIC TRACHEA II-65 418-028:PAGE J-96 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMETNOTXAILCITYSCREENINGTEST PROTOCONLUMBER416-028 SPONSOR'SSTUDYNUMBERT-7706.1 Appendix II IndividualAnimal 6ross and HistomorphologyData ANIMAL NUMBER: 19025 DOSE GROUP: SEX: F DEATH TYPE: ======================================================================================================= V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: ADRENALGLANDS: hypertrophy/vacuolationc,ortex,multifocal (mild) KIDNEYS: nephritis,chronic,focal (minimal) STOMACH: edema/inflammations,ubmucosa,glandulararea (mild) UTERUS: macrophages,pigmented(moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: BONE MARROW (STERNUM) DUODENUM BRAIN HEART CECUM ILEUM LIVER MAMMARYGLAND LUNG NERVE, SCIATIC LYMPH NODE, MEDIASTINAL OVARIES PEYER'SPATCH RECTUM SPINAL CORD, CERVICAL SPINALCORD, THORACIC TRACHEA SPLEEN URINARYBLADDER THYMUS VAGINA ....................................................................................................... End of Record- 19025 COLON JEJUNUM LYMPH NODE, SUBMANDIBULAR PARATHYROID SPINAL CORD, LUMBAR THYROID II-66 418-028:PAGE J-97 ORAL {GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-770B.1 ANIMAL NUMBER: 19027 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOL061COBSERVATION(S); GENERAL_L:Ngoross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: KIDNEYS: mineralization,multifocal (minimal) LYMPH NODE, SUBMANDIBULAR: hyperplasia,lymphoeytic/plasmacyti(cmild) STOMACH: edema/infla_ation,submuco_a,glandulararea (mild) THYROID: ultimobranchialbody/cyst ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS BONE MARROW (STERNUM) BRAIN COLON DUODENUM HEART JEJUNUM LIVER LUNG MAMMARY6LAND OVARIES PARATHYROID RECTUM SPINAL CORD, CERVICAL SPINAL CORD, LUMBAR SPLEEN THYMUS TRACHEA UTERUS VAGINA ....................................................................................................... CECUM ILEUM LYMPH NODE, MEDIASTINAL PEYER'SPATCH SPINALCORD, THORACIC URINARY BLADDER TISSUE(S)NOT AVAILABLEFOR EVALUATION: NERVE, SCIATIC ....................................................................................................... End of Record- 19027 11-67 418-028:PAGE J-98 ORAL {GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-7706.I Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 1902B SEX: F l s DosE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): GENERAL:No gross changes. Not applicable ...................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LYMPH NODE, SUBMANDIBULAR: STOMACH: hyperplasia,lymphocytic/plasmacyti(cmoderate) dilatation,mucosal glands (minimal) UTERUS: macrophages,pigmented (moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COLON JEJUNUM BONE MARROW (STERNUM) DUODENUM KIDNEYS BRAIN HEART LIVER LYMPH NODE, MEDIASTINAL PARATHYROID SPINAL CORD, LUMBAR THYROID MAMMARY GLAND PEYER'S PATCH SPINAL CORD, THORACIC TRACHEA NERVE, SCIATIC RECTUM SPLEEN URINARYBLADDER ....................................................................................................... End of Record- 19028 CECUM ILEUM LUNG OVARIES SPINAL _ORD, CERVICAL THYMUS VAGINA II-B8 418-028:PAGE J-99 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-770B WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 41B-028 SPONSOR'SSTUDY NUMBER T-770B.I ANIMAL NUMBER: 19030 SEX: F Appendix II IndividualAnimal Gross and HistomorphologyData DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOGICOBSERVATION(S): HEAD."Head- scab, O.3cm x O.3cm. dermatitis,chronic,focal, SKIN (GROSSLESION) ....................... ................................................................................ HISTOMORPHOLOGICOBSERVATIONS: LIVER: Not remarkable SKIN (GROSS LESION): dermatitis,chronic,focal (minimal) THYROID: ultlmobranchialbody/cyst UTERUS: macrophages,pigmented (moderate) ....................................................................................................... THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS COLON BONE MARROW (STERNUM) DUODENUM BRAIN HEART JEJUNUM MAMMARYGLAND KIDNEYS NERVE, SCIATIC LUNG OVARIES PEYER'S PATCH SPINAL CORD, THORACIC TRACHEA RECTUM SPLEEN URINARY BLADDER SPINALCORD, CERVICAL STOMACH VAGIRA ....................................................................................................... CECUM ILEUM LYMPH NODE, MEDIASTINAL PARATHYROID SPINAL CORD, LUMBAR THYMUS TISSUE(S)NOT AVAILABLEFOR EVALUATION: LYMPH NODE, SUBMANDIBULAR ....................................................................................................... End of Record- 19030 II-B9 418-028:PAGE J- 100 ORAL (GAVAGE)COMBINEDREPEATEDDOSE TOXICITYSTUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTATLOXICITYSCREENINGTEST PROTOCOLNUMBER 418-028 SPONSOR'SSTUDY NUMBER T-77OB.1 Appendix II IndividualAnimal Gross and HistomorphologyData ANIMAL NUMBER: 19031 SEX: F DOSE GROUP: DEATH TYPE: V Sacrifice-Scheduled GROSS OBSERVATION(S): HISTOMORPHOLOBICOBSERVATION(S): GENERAL:No gross changes. Not applicable ....................................................................................................... HISTOMORPHOLOGICOBSERVATIONS: LIVER: LYMPH NODE, SUBMANDIBULAR: UTERUS: necrosis,focal (minimal) hyperplasia,lymphocytic/plasmacyti(cmild) distention,lumen (moderate) THE FOLLOWINGTISSUE(S)WERE WITHIN NORMAL LIMITS: ADRENALGLANDS BONE MARROW (STERNUM) BRAIN COLON DUODENUM HEART JEJUNUM KIDNEYS LUNG MAMMARYGLAND PEYER'SPATCH NERVE, SCIATIC RECTUM OVARIES SPINAL CORD, CERVICAL SPINAL CORD, THORACIC THYROID SPLEEN TRACHEA STOMACH URINARYBLADDER ....................................................................................................... End of Record- 19031 CECUM ILEUM LYMPH NODE, MEDIASTINAL PARATHYROID SPINAL CORD, LUMBAR THYMUS VAGINA II-/O 418-028:PAGE J- 101 ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL 418-028 SPONSOR'S STUDY NUMBER T-7706.1 HISTOPATHOLOGY REPORT APPENDIX III HISTOMORPHOLOGIC OBSERVATIONS IN THE LIVER F1 GENERATION PUPS 418-028:PAGE J- 102 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF T-7706 WiTH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Table II1-1 HistomorphologiOc bservationsin the Liver- GroupI - F1GenerationPups DoseGroup: DamNumber: PupNumber: LIVER." I I I I I I I I I I 19012 190121901219012 190121901219012 1901219012 19012 1 2 3 4 5 7 8 14 15 16 DoseGroup: DarnNumber. PupNumber. LIVER." I I I I I I I I I I 19019 190191901919019 190191901919019 1901919019 19019 1 2 3 7 8 11 12 13 14 15 " "" DoseGroup: DamNumber. PupNumber: LIVER: I I I I I I I ! I I 19021 190211902119021 1902119021 1902119021 1902119021 1 2 6 8 9 10 11 12 13 14 DoseGroup: DamNumber: PupNumber: LIVER: I I I I I I I I I I 19023 19023190231902319023 1902319023 1902319023 19023 1 2 3 6 7 8 9 10 11 12 - - DoseGroup: DamNumber. PupNumber: LIVER: I I I I I I I I I I 1904119041 190411904119041 190411904119041 1904119041 1 2 3 4 5 8 9 15 16 17 II1-1 418-028 :PAGE J- 103 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 TableII1-1(Continued) HistomorphologiOc bservalJonisn the Liver- GroupI - F1 GenerationPups DoseGroup: DamNumber: PupNumber: LIVER: I I I I I I I I I I 19042 1904219042 190421904219042 1904219042 1904219042 1 2 4 5 6 7 8 10 12 13 - - - DoseGroup: DamNumber: PupNumber: LIVER: I I I I I I I I I I 19044 190441904419044 190441904419044 1904419044 19044 1 2 5 6 7 9 11 12 13 14 DoseGroup: DamNumber: PupNumber: LIVER: I I I I I I I I I I 1905019050 19050190501905019050 1905019050 1905019050 1 2 3 4 5 6 7 8 13 14 - - DoseGroup: DamNumber: PupNumber. LIVER: DoseGroup: DamNumber: PupNumber:. LIVER: I I I I I I I I I I 190531905319053 190531905319053 190531905319053 19053 1 3 4 5 6 10 11 12 13 14 - - - I I I I I I I I I I 19065 190651906519065 190651906519065 190651906519065 3 4 5 7 8 "10 11 14 15 17 111-2 418-028:PAGE J- 104 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF "I-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Table 111-2 HistomorphologOicbservationinstheLiver-GroupV- F1GenerationPups DoseGroup: DamNumber. PupNumber: LIVER: VVVVVVVVV 190011900119001 1900119001 190011900119001 19001 1 2 3 4 5 6 7 9 11 - - DoseGroup: DamNumber: PupNumber:. LIVER: V VVVVVVVVV 1900619006 1900619006 190061900619006 1900619006 19006 1 2 3 4 5 8 10 11 13 18 - - DoseGroup: DamNumber: PupNumber: LIVER: V VVVVVVVVV 190111901119011 1901119011 190111901119011 1901119011 2 3 4 5 6 9 10 11 12 13 - - DoseGroup: DamNumber:. PupNumber:. LIVER: VVVVVVVVVV 190201902019020 190201902019020 1902019020 1902019020 2 4 5 7 8 9 10 11 12 13 - - - DoseGroup: DamNumber: PupNumber:. LIVER: VVVVVVVVVV 19022 190221902219022 190221902219022 1902219022 19022 1 4 5 7 8 11 12 13 !4 15 - - 111-3 418-028:PAGE J-105 ORAL (GAVAGE) COMBINED REPEATED DOSE STUDY OF T-7706 WITH THE REPRODUCTION/DEVELOPMENTAL TOXICITY SCREENING TEST PROTOCOL NUMBER 418-028 SPONSOR'S STUDY NUMBER T-7706.1 Table111-(2Continued) HistomorphologOibcservationisntheLiver- GroupV- F1 GeneratioPnups DoseGroup: DamNumber: PupNumber:. LIVER: V VVVVVVVVV 190251902519025 190251902519025 1902519025 1902519025 1 2 7 8 13 14 15 18 19 20 - - DoseGroup: DamNumber: PupNumber: LIVER: VVVVVVVVV 190271902719027 190271902719027 1902719027 19027 1 2 3 4 5 6 7 13 14 DoseGroup: DamNumber: PupNumber: LIVER: V VVVVVVVVV 1902819028 19028190281902819028 1902819028 1902819028 1 2 3 4 5 6 7 10 11 12 - - DoseGroup: DamNumber: PupNumber. LIVER: VVVVVVVVVV 190301903019030 190301903019030 190301903019030 19030 1 2 4 5 6 8 9 11 12 13 - - - DoseGroup: DamNumber. PupNumber:. LIVER: VVVVVVVVVV 19031 1903119031 19031190311903119031 1903119031 19031 1 2 3 5 6 9 11 12 14 15 - - 111-4 APPENDIX K HEMATOLOGY AND CLINICAL CHEMISTRY REPORTS 418-028:PAGE K-1 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID MBC THSN/CU RM RBC MILL/CU RN GROUP: I-M 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 19143 19151 19157 19161 19167 13.5 15.9 15.1 12.7 16.4 17.6 17.7 14.9 15.0 14.3 ........ ........ ........ ........ ........ 7.10 8.58 6.95 7.16 6.7"7 7.61 7.37 7.45 7.43 8.71 MEAN SD N 15.3 1.63 10 7.51 0.648 10 HGB GRARS/DL 15.4 19.0 16.3 16.2 15.7 16.2 16.3 16.3 15.6 18.2 16.5 1.16 10 SEX: MALE HCT % NCV CU MICRONS MCH PICO GRAMS MCHC % PLT THSN/CU MM 40.6 50.0 42.6 42.1 40.6 41.4 41.6 43.3 42.7 /,9.7 57.2 58.3 61.3 58.8 59.9 54.4 56.5 58.1 57.5 57.I 21.7 22.1 23.5 22.6 23.2 21.3 22.1 21.9 21.0 20.9 37.9 38.0 38.3 38.5 38.7 39.I 39.2 37.6 36.5 36.6 1219 1072 964 1354 1074 1322 1049 1264 1269 1203 43.5 3.48 10 57.9 1.89 10 22.0 0.87 10 38.0 0.93 10 1179 130.9 10 (--) - Data Unavai LabLe LABCAT HE4.43 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-2 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: MALE Animal ID _C THSN/CU MM _C MILL/CU 191 GROUP: 2-M 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 19147 19153 19164 19165 19171 10.9 7.55 17.0 6.9"5 12.7 7.02 _ 13.1 8.20 15.7 6.80 18.4 7.42 14.7 7.29 10.5 7.32 17.2 7.53 19.0 7.20 .......... ........ ......... ........ ........ REAN SO N 14.9 3.03 10 7.3"5 0.396 10 _B GRAMS/DL 16.2 15.2 16.1 15.9 16.1 16.4 15.9 15.4 16.4 15.5 15.9 0.42 10 HCT MCV RCH % CU MICRONS PICO GRARS 43.6 39.6 40.3 43.9 42.0 44.4 43.2 41.3 42.2 41.7 57.7 57.1 57.4 53.5 61.8 59.8 59.3 56.4 56.0 57.9 21.5 21.9 22.9 19.4 23.7 22.1 21.8 21.0 21.8 21.5 MCHC % PLT THSN/CU MS 37.2 38.4 40.0 36.2 38.3 56.9 36.8 37.3 38.9 37.2 926 970 959 1468 1170 1382 1184 1183 1069 994 42.2 1.57 10 57.7 2.27 10 21.8 1.13 10 37.7 1.15 10 1131 183.7 10 (--) - Data UnavailabLe LABCAT HE4.43 27-JUN-2(X)2 418-028:PAGE K-3 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS418-028 STUDY NO: 060-069 Animal ID WBC THSN/CU MM RBC MILL/CU MR GROUP: 3-M 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 19155 19159 19172 19173 19174 13.1 11.0 12.5 13.3 11.2 20.9 14.1 13.9 12.2 15.4 ......... ........ ........ ........ ........ 7.38 7.28 7.59 7.20 7.72 7.23 6.69 7.96 7.60 6.54 MEAN SD N 13.8 2.84 10 7.32 0.441 10 HGB GRAMS/DL 16.0 16.3 16.1 15.4 15.7 15.4 15.0 16.4 16.3 14.7 15.7 0.59 10 SEX: MALE HCT % MCV CU MICRONS MCH PICO GRAMS MCHC % PLT THSN/CU MR 42.4 43.2 /-,4.4 40.1 42.6 40.5 39.1 43.8 45.0 38.8 57.4 59.3 58.5 55.7 55.2 56.0 58.4 55.0 59.2 59.4 21.7 22.4 21.2 21.4 20.3 21.3 22.4 20.6 21.4 22.5 37.7 37.7 36.3 38.4 36.9 38.0 38.4 37.4 36.2 37.9 958 960 993 1002 1253 1340 1143 1125 1046 1336 42.0 2.22 10 57.4 1.78 10 21.5 0.75 10 37.5 0.79 10 1115 149.2 10 (--) - Data Unavailable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-4 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID WBC THSN/CU MR RBC MILL/CU MM GROUP: 4-M 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 19156 19160 19162 19163 19166 12.6 6.59 21.0 6.73 18.1 6.75 12.0 6.83 22.0 7.50 17.4 7.07 12.8 6.87 19.6 7.37 15.6 6.97 11.1 6.62 .......... ......... ........ ........ ......... MEAN SD N 16.2 3.97 10 6.93 0.305 10 HGB 6RAMS/DL 14.6 15.0 15.6 14.8 16.8 15.5 15.0 16.1 15.6 15.0 15.4 0.67 10 HCT % MCV U MICRONS MCH PICO GRAMS 38.2 39.0 39.3 38.4 45.5 40.3 39.9 42.4 40.6 38.1 57.9 58.0 58.2 56.2 60.7 57.0 58.1 57.5 58.2 57.6 22.2 22.3 23.1 21.7 22.4 21.9 21.8 21.8 22.4 22.7 40.2 2.29 10 57.9 1.15 10 22.2 0.45 10 SEX: MALE MCHC % PLT THSN/CU NM 38.2 38.5 39.7 38.5 36.9 38.5 37.6 38.0 38.4 39.4 1072 1144 1029 1274 1124 1125 998 962 1155 1161 38.4 0.80 10 1104 91.4 10 (--) - Data UnavailabLe LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-5 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID WBC THSN/CU MR RBC MILL/CU MR GROUP: 5-M 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 19154 19158 19168 19169 19170 15.7 7.20 14.3 6.72 10.8 6.68 12.5 8.07 11.0 6.61 14.2 7.09 15.2 7.05 20.7 7.13 18.3 6.66 13.9 6.68 ......... ......... .......... ........ ......... MEAN SO N 14.7 3.07 10 6.99 0.442 10 HGB GRAMS/DL 16.1 15.7 15.0 17.3 14.7 15.5 16.2 15.7 14.8 14.8 15.6 0.81 10 HCT % MCV CU MICRONS NCfl PICO GRAMS 41.7 39.8 39.0 44.4 38.1 41.1 41.9 41.4 39.8 39.5 57.9 59.2 58.4 55.0 57.6 58.0 59.5 58.1 59.8 59.2 22./+ 23.4 22.5 21.4 22.2 21.9 23.0 22.0 22.2 22.2 40.7 1.81 10 58.3 1.37 10 22.3 0.56 10 SEX: MALE MCHC % PLT THSN/CU MN 38.6 39.4 38.5 39.0 38.6 37.7 38.7 37.9 37.2 37.5 1013 1181 1249 1142 1040 1001 1059 1523 1234 1041 38.3 0.70 10 1148 159.9 10 (--) - Data UnavaiLabLe LABCAT HE4.43 27-JUN-2002 418-028:PAGE K6 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060--069 Animal ID WBC THSN/CU MM RBC MILL/CU MM GROUP: 1-F 19010 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 19074 19075 ......... 15.9 6.6 7.4 7.3 8.0 6.8 10.6 8.2 6.6 12.5 ........ ........ ........ ........ 5.60 6.95 7.10 6.62 6.86 6.45 6.09 6.64 6.08 6.67 MEAN SD N 9.0 3.09 10 6.51 0.461 10 HGB GRARS/DL 14.0 16.4 16.7 16.3 16.2 15.1 14.3 16.0 14.7 15.7 15.5 0.95 10 HCT % MCV CU MICRONS MCH PICO GRAMS 35.2 42.5 44.7 42.4 42.4 40.2 37.6 42.2 37.5 40.2 62.9 61 .I 63.0 64.1 61.8 62.3 61.7 63.6 61.6 60.2 25.0 23.6 23.5 24.6 23.6 23.4 23.5 24.1 24.2 2.'3.5 40.5 2.93 10 62.2 1.19 10 23.9 0.55 10 SEX: FEMALE fiche % PLT THSN/CU MM 39.8 38.6 37.4 38.4 38.2 37.6 38.0 3-7.9 59.2 39.1 1421 1344 1199 1352 1529 1225 1343 1629 1524 1623 38.4 0.76 10 1419 153.0 10 (--) - Oalca Unavailable LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-7 Study Report for Hematology INDIVIDUAL ANIMAL RRPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID WBC THSN/CU MM RBC MILL/CU MM GROUP: 2-F 19004 19009 19016 19018 19026 19036 19037 19043 19047 19048 19052 19055 19061 19067 19071 7.8 14.6 17.0 13.3 9.1 18.0 26.3 13.5 11 .I 7.1 ......... ........ ........ ......... ......... 7.07 5.90 6.32 5.61 6.36 6.30 6.23 5.83 5.93 6.60 MEAN SO N 13.8 5.73 10 6.22 0.423 10 HGB GRAMS/DL 16.6 1&.8 14.9 14.1 15.5 14.5 14.8 14.7 14.5 16.0 15.0 0.77 10 HCT % MCV CU MICRONS MCH PICO GRAMS 43.8 38.3 38.5 35.3 41.6 38.9 37.8 38.3 37.5 43.4 62.0 64.9 60.9 63.0 65.4 61.7 60.6 65.7 63.3 65.8 23.5 25.1 23.6 25.1 24.4 23.0 23.8 25.2 24.5 24.2 SEX: FEMALE MCHC % PLT THSN/CU MM 37.9 38.6 38.7 39.9 37.3 37.3 39.2 38.4 38.7 36.9 1540 1157 1529 1057 1443 1166 1257 942 1041 1264 39.3 2.72 10 63.3 2.01 10 24.2 0.76 10 38.3 0.94 10 1240 208.1 10 (--) - Data UnavaiLable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-8 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD: TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID WBC THSN/CU MM RBC MILL/CU MM GROUP: 3-F 19003 19007 19008 19013 19014 19015 19017 19024 19029 19034 19038 19056 19057 19060 19064 11.1 6.43 7.9 6.31 7.0 6.71 16.6 5.53 ......... 7.4 6.&I 12.9 6.03 10.5 6.93 12.5 5.69 12. I 6.54 .......... 12.7 7.38 .......... ......... ......... MEAN SD N 11 .I 2.98 10 6.40 0.554 10 HGB GRAMS/DL 15.7 15.4 15.7 13.8 15.1 14.4 15.6 14.0 16.1 16.6 15.2 0.91 10 HCT % MCV CU MICRONS MCH PICO GRAMS 40.0 39.9 40.5 33.1 39.8 38.0 42.1 35.6 41.9 44.2 62.2 63.3 60.3 59.9 62.1 63.0 60.8 62.6 64.I 59.9 24_4 24.4 23.4 25.0 23.6 23.9 22.5 24.6 24.6 22.5 SEX: FEMALE MCHC % PLT THSN/CU RR 39.3 38.6 38.8 41.7 37.9 37.9 37.1 39.3 38.4 37.6 1165 1450 1319 1167 1671 B66 1424 1062 1231 1380 39.5 3.24 10 61.8 1.50 10 23.9 0.88 10 38.7 1.29 10 1274 226.3 10 (--) - Data Unavailable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-9 Study INDIVIDUAL Report for Hematology ANIMAL PERIOD : R_-PORT TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID WBC THSN/CU RM RBC MILL/CU RM GROUP: 4-F 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058 19062 19063 19066 19069 19073 ......... 9.5 6.51 9.6 6.81 12.8 6.48 I0.0 9.7 6.48 5.77 ........ .......... 14.9 7.33 15.7 5.94 6.2 6.86 12.9 5.68 7.1 6.01 ........ ........ MEAN SD N 10.8 3.15 10 6.39 0.532 10 HGB GRAMS/DL 15.8 16.2 14.8 15.8 13.7 15.7 14.2 15.7 14.0 15.7 15.2 0.90 10 HCT % MCV CU MICRONS MCH PICO GRAMS 42.0 42.4 38.9 41.3 36.7 44.1 37.2 43.3 35.2 40.3 64.5 62.3 60.1 65.8 63.6 60.2 62.6 63.1 61.9 67.0 24.3 23.8 22.8 24.4 23.7 21.4 23.9 22.9 24.6 26.1 40.1 3.02 10 62.9 2.04 10 23.8 1.25 10 SEX: FEMALE MCHC % PLT THSN/CU Mff 37.6 38.2 38.0 38.3 37.3 35.6 38.2 36.3 39.8 39.0 1479 1527 1066 1333 1148 1497 1094 1399 999 1506 37.8 1.22 10 1305 207.2 10 (--) - Data Unavailable LABCAT HE4.43 27-JUN-2002 Study INDIVIDUAL Report for Hematology ANIMAL PERIOD : REPORT TERMINAL BY GROUP 418-028:PAGE K-10 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID WBC THSN/CU _ RBC MILL/CU Mfl GROUP: 5-F 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 15.1 5.90 7.6 6.33 6.4 6.46 4.2 7.49 5.4 6.79 7.3 6.47 12.3 6.69 10.2 5.76 8.1 6.62 12.3 5.72 .......... ........ ........ ......... ........ MEAN SD N 8.9 3.48 10 6.42 0.538 10 HGfi GRAMS/DL 14.3 15.4 15.7 16.1 16.0 15.I 15.9 13.8 15.6 14.7 15.3 0.77 10 SEX: FEMALE HCT % RCV CU MICRONS MCH PICO GRAMS 37.3 39.0 40.5 46.8 43.0 39.7 42.2 35.8 41.4 38.4 63.2 61.6 62.7 62.5 63.4 61.3 63.1 62.2 62.6 67.1 24.2 24.3 24.3 21.5 23.6 23.3 23.8 24.0 23.6 25.7 MCHC % PLT THSN/CU 38.3 39.5 38.8 34.4 37.2 38.0 37.7 38.5 37.7 38.3 1131 1312 1254 1549 1448 1391 1667 1286 1417 1099 40.4 3.15 10 63.0 1.60 10 23.8 1.05 10 37.8 1.37 10 1355 176.8 10 (--) - Data UnavaiLable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-11 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID MPV CU MICRON PT seconds GROUP: q-M 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 19143 19151 19157 19161 19167 8.0 13.2 8.0 13.3 7.2 13.7 7.5 13.6 8.2 13.1 8.9 13.6 8.2 13.2 8.0 13.7 8.4 13.2 8.8 ........ 13.2 ......... ......... ........ -- -- MEAN SD N 8.1 0.52 10 13.4 0.24 10 APTT seconds 13.2 21.5 22.4 23.0 19.8 20.8 19.8 21.8 23.9 23.6 " 21.0 3.09 10 SEX: MALE NRBC COUNT Lymphocyte THSN/CU MR Segmented THSN/CU MM Bands THSN/CU NM Ronocytes THSN/CU MM 0 9.7 2.0 0.0 1 .I 0 11.4 2.7 0.0 1.6 0 11.6 3.0 0.0 0.3 0 9.1 2.2 0.0 1.3 0 14.1 1.5 0.0 0.2 0 14.3 1.4 0.0 1 .I 0 13.1 2.7 0.0 1.6 0 12.7 1.5 0.0 0.4 0 11.0 3.5 0.0 0.5 0 12.2 1.6 0.0 0.3 ..... 0 0.0 10 11.9 1.72 10 2.2 0.73 10 0.0 0.00 10 0.8 0.56 10 (--) - Data Unavailable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-12 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 AnimaL ID MPV CU MICRON PT seconds GROUP: 2-M 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 19147 19153 19164 19165 19171 7.5 14.0 7.6 14.0 8.4 14.6 8.4 14.2 7.2 14.4 7.6 14.4 7.2 14.3 8.8 14.2 8,1 13.7 7.8 14.5 ......... .......... .......... ........ ........ MEAN SD N 7.9 0.54 10 14.2 0.27 10 APTT seconds 22.1 22.0 24.6 26.2 20.0 17.5 20.8 19.3 16.6 31.4 22.1 4.41 10 SEX: MALE NRBC COUNT Lymphocyte THSN/CU MM Segmented THSN/CU MM Bands THSN/CU MR Monoc)rt es THSN/CU NM 0 6.5 2.7 0.0 1.0 0 12.1 2.9 0.0 1.4 0 10.9 1.4 0.0 0.1 0 10.0 2.0 0.0 0.7 0 11.1 3.6 0.0 0.8 0 13.4 3.1 0.0 1.5 0 9.4 4.0 0.0 0.7 0 8.6 1.4 0.0 0.3 0 11.4 5.2 0.0 0.2 0 16.3 2.5 0.0 0.2 0 11.0 2.9 0.0 0.7 Q.O 2.68 1.18 0.00 0.50 10 10 10 10 10 (--) - Data UnavaiLabLe LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-13 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID RPV CU MICRON PT seconds GROUP: 3-M 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 19155 19159 19172 19173 19174 8.0 7.4 8.0 9.9 8.3 8.3 8.1 8.9 7.8 7.6 ......... ........ -........ ........ 13.9 13.7 13.6 13.6 13.5 13.2 13.7 13.7 13.4 14.0 -- MEAN SO N 8.2 0.72 10 13.6 0.23 10 APTT seconds 18.3 24.6 2.2.0 22.1 19.8 21.2 18.1 22.3 20.6 16.7 20.6 2.38 10 SEX: MALE NRBC COUNT Lymphocyte THSN/CU RR Segmented THSN/CU MM Bands THSN/CU MR Monocytes THSN/CU HM 0 10.1 2,1 0.0 0.5 0 8.8 1.9 0.0 0.2 0 9.8 0.6 0.1 1.5 0 11.0 1.3 0.0 0.5 0 7.8 2.7 0.0 0.6 0 14.8 4.0 0.0 1.5 0 10.4 3.4 0.0 0.1 0 11.7 1.7 0.0 0.4 0 10.6 1.1 0.0 0.2 0 13.1 1.4 0.0 0.3 0 10.8 2.0 0.0 0.6 0.0 2.02 1.06 0.03 0.51 10 10 10 10 10 (--) - Data UnavailabLe LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-14 study Report for Hematology INDIVIDUAL ANIMAL PERIOD: REPORT TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID MPV CU MICRON PT seconds GROUP: 4-M 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 19156 19160 19162 19163 19166 7.1 8.2 7.9 8.3 7.4 8.4 7.5 9.3 8.7 9.3 ......... ......... ........ ......... ......... 13.5 14.6 14.0 13.9 14.0 13.6 13.5 13.9 13.8 13.1 MEAN SD N 8.2 0.76 10 13.8 0.40 10 APTT seconds 19.8 26.3 23.0 21.4 19.1 21.5 20.8 21.5 19.7 18.6 21.2 2.24 10 SEX: MALE NRBC COUNT Lymphocyte THSN/CU tim Segmented THSN/CU MM Bands THSN/CU MM Monocyt es THSN/CU MR 0 7.7 4.0 0.0 0.6 0 16.4 2.3 0.0 1.7 0 12.1 3.1 0.0 2.2 0 9.4 1.4 0.0 0.5 0 17.4 2.9 0.0 0.7 0 12.7 3.7 O. 0 O. 5 0 8.6 2.8 0.1 1.0 0 15.9 2.7 0.0 0.6 0 12.3 3.1 0.0 0.0 0 8.3 2.4 0.0 0.2 0 12.1 2.8 0.0 0.8 0.0 3.57 0.73 0.03 0.67 10 10 10 10 10 (--) - Data Unavailable LABCAT HE4.43 27-JUN-2002 Study INDIVIDUAL Report for Hematology ANIMAL PERIOD: REPORT TERMINAL BY GROUP 418-028:PAGE K-15 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 AnimaL ID NPV CU RZCRON PT seconds GROUP: 5-M 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 19154 19158 19168 19169 19170 7.7 7.5 7.3 9.0 8.8 8.6 8.0 7.5 7.8 8.2 ........ ......... ......... ........ ......... 13.9 15.1 13.8 13.7 14.1 13.5 13.9 14.3 13.5 13.7 MEAN SD N 8.0 0.59 10 14.0 0.47 10 APTT seconds 23.6 23.3 20.8 21.8 20.7 21.6 19.6 21.9 19.5 20.0 21.3 1.43 10 SEX: MALE NRBC COUNT Lymphocyte THSN/CU MR Segmented THSN/CU MR Bands THSN/CU MR HonocyZes THSN/CU MM 0 13.0 1.4 0.0 0.6 0 11,3 1.7 0.0 0.9 0 10.2 0.3 0.0 0.2 0 10.1 1.1 0.0 0.9 0 8.0 2.4 0.0 0.2 0 12.4 1.4 0.0 0.3 0 12.2 2.1 0.0 0.2 0 17.2 2.5 0.0 0.8 0 16.1 2.2 0.0 0.0 0 13.1 0.4 0.0 0.4 0 12.4 1.6 0.0 2.75 0.78 10 10 10 0.0 0.00 10 0.5 0.33 10 (--) - Data UnavaiLable LABCAT HE4.43 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-16 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal TO MPV CU MICRON PT seconds GROUP: 1-F 19010 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 19074 19075 .......... 7.7 13,9 8.9 13.0 7.8 13.8 7.9 12.9 9.1 13.4 8.4 12.8 8.4 12.8 8.2 13.1 8.1 13.3 8.9 12.8 .......... ........ ........ .......... MEAN SD N 8.3 0.49 10 13.2 0.41 10 APTT seconds 26.6 20.4 24.2 21.9 19.3 21.8 18.5 20.5 22.5 21.6 21.7 2.36 10 SEX: FEMALE NRBC COUNT Lymphocyte THSN/CU 1_ Segmented THSN/CU MM Bands THSN/CU MR Monocytes THSN/CU MM 0 12.6 2.7 0.0 0.5 0 4.4 2.0 0,0 0.2 0 5,7 1.4 0.0 0.0 0 6.1 0.9 0.0 0.1 0 5.0 2.9 0.0 0.1 0 5.3 1.5 0.0 0.0 0 7.3 3.1 0.0 0.1 0 6.1 1.8 0.0 0.2 0 4.7 1.8 0.0 0.0 0 8.4 3,8 0.0 0.1 0 6.6 0.0 2.44 10 10 2.2 0.90 10 0.0 0.00 10 0.1 0.15 10 (--) - Data Unavailable LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-17 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE Animal ID MPV CU MICRON PT seconds GROUP: 2-F 19004 19009 19016 19018 19026 1905 19037 19043 19047 19048 19052 19055 19061 19067 19071 9.6 13.3 7.2 13.8 8.8 13.3 " 8.2 13.7 8.4 12.8 7.6 13.5 8.1 13.3 8.0 13.2 6.9 13.5 7.0 13.1 ............ ......... ........ ........ ........ MEAN SD N 8.0 0.84 10 13.4 0.29 10 APTT seconds 27.5 21.3 29.2 22.6 20.3 24.3 20.0 26.8 25.3 22,1 23.9 3.19 10 NRBC COUNT Lymphocyte THSN/CU MR Segmented THSN/CU MR Bands THSN/CU MM nonocyt es THSN/CU MM 0 5.8 2.0 0.0 0 12.7 1.6 0.0 0 13.6 3.1 0,0 0 10.9 2.3 0.0 0 7.7 1.4 0.0 0 15.1 2.2 0.0 0 11.0 13.7 0.0 0 9.6 3.5 0.0 0 6.2 4.6 0.0 0 5.4 1.6 0.0 0.1 O. I 0.0 0.0 0.0 0.4 1.6 0.3 0.2 0.0 0 9.8 3.6 0.0 0.3 0.0 3.44 3.69 0.00 0.49 10 10 10 10 10 (--) - Data UnavaiLabLe LABCAT HE4.43 27- JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD: TERMINAL BY GROUP 418-028:PAGE K-18 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID MPV CU MICRON PT seconds GROUP: 3-F 19003 19007 19008 19013 19014 19015 19017 19024 19029 19034 19038 19056 19057 19060 19064 7.2 12.9 7.3 12.9 9.3 12.8 8.5 13.7 .......... 9.0 13.6 7.8 13.3 8.8 12.9 7.4 13.3 8.1 ........ 12.7 9.3 13.6 ......... ........ ......... MEAN SD N 8.3 0.82 10 13.2 0.37 10 APTT seconds 22.7 21.7 22.6 22.7 27.6 21.9 22.1 24.4 21.2 27.6 23.5 2.35 10 SEX: FEMALE NRBC COUNT Lymphocyte THSN/CU MR Segmented THSN/CU RM Bands THSN/CU MM Monocytes THSN/CU MM 0 6.5 4.0 0.0 0.3 0 5.7 2.0 0.0 0.2 0 5.0 1.9 0.0 0.0 0 12.3 3.8 0.0 0.3 0 5.7 1.6 0.0 0.1 0 9.8 2.6 0.0 0.1 0 7.4 2.6 0.0 0.2 0 10.3 2.1 0.0 0.0 0 10,9 1.0 0.0 0.1 0 10.2 2.2 0.0 0.4 0 8.4 2.4 0.0 0.2 0.0 2.60 0.93 0.00 0.13 10 10 10 10 10 (--) - Data Unavailable LABCAT HE4.43 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028 :PAGE K- 19 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID MPV CU MICRON PT seconds GROUP: 4-F 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058 19062 19063 19066 19069 19073 .......... 7.8 13.1 9.0 13.4 7.8 13.4 8.1 12.9 7.6 13.0 .......... ........ 7.5 14.1 7.5 13.5 9.1 13.9 8.5 14.0 7.4 13.5 ........ ........ MEAN SD N 8.0 0.63 10 13.5 0.42 10 APTT seconds 21.4 24.5 25.7 27.9 21.9 27.1 25.7 25.2 26.2 28.9 25.3 2.45 10 SEX: FEMALE NRBC COUNT Lymphocyte THSN/CU HH Segmented THSN/CU MM Bands THSN/CU MR Honocytes THSN/CU NM 0 7.7 1.6 0.0 0.1 0 7.1 2.3 0.0 0.1 0 9.6 2.8 0.0 0.3 0 7.7 2.2 0.0 0.1 0 5.4 3.8 0.3 0.2 0 12.2 1.9 0.0 0.4 0 11.5 3.9 0.0 0.2 0 4.8 1.4 0,0 0.1 0 9.8 2.6 0.0 0.3 0 6.0 1.0 0.0 0.1 0 8.2 2.4 0.0 0.2 0.0 2.52 0.96 0.09 0.11 10 10 10 10 10 (--) - Data Unavailable LABCAT HE4.43 27-J UN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD ; TERMINAL BY GROUP 418-028 :PAGE K-20 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID MPV CU MICRON PT seconds GROUP: 5-F 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 6.9 13.8 9.2 12.8 7.4 12.9 7.0 16.8 7.6 13.7 8.3 13.5 7.3 13.2 8.1 13.6 7.9 13.7 6.8 13.5 ......... ......... .......... ......... .......... MEAN SD N 7.7 0.74 10 13.8 1.12 10 APTT seconds 30.4 22.1 22.3 27.6 21.6 25.3 27.5 22.2 31.5 21.6 25.2 3.81 10 SEX: FERALE NRBC COUNT Lymphocyte THSN/CU MN Segmented THSN/CU MM Bands THSN/CU MM Monocyt es THSN/CU MM 0 10.1 4.2 0.0 0.5 0 4.6 2.7 0.0 0.2 0 4.4 1.8 0.0 0.1 2 2.2 1.9 0.0 O. 1 0 3.1 2.0 0.0 0.2 0 5.2 2.0 0.0 0.1 0 10.2 2.0 0.0 0.1 0 7.1 2.8 0.0 0.3 0 5.2 2.8 0.0 0.1 0 8.2 3.7 0.0 0.0 0 6.0 2.6 0.0 0.2 0.6 2.77 0.82 0.00 0.14 10 10 10 10 10 (--) - Data UnavaiLable LABCATHE4.43 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-21 SEX: MALE Animal ID Eosinophil THSN/CU MR Basophils THSN/CU MM Abnormal L THSN/CU MR Other THSN/CU MM GROUP: 1-M 19109 0.7 0.0 0.0 0.0 19115 0.2 0.0 0.0 0.0 19116 0.2 0.0 0.0 0.0 19119 0.1 0.0 0.0 0.0 19122 0.2 0.0 0.5 0.0 19125 0.5 0.0 0.4 0.0 19131 0.2 0.0 0.2 0.0 19132 0.0 0.0 0.3 0.0 19134 0.2 0.0 0.0 0.0 19135 0.3 0.0 0.0 0.0 19143 .... 19151 .... 19157 ..... 19161 .... 19167 .... MEAN SD N 0.3 0.20 10 0.0 0.00 10 0.1 0.20 10 0.0 0.00 10 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-22 SEX: MALE Animal ID Eosinophil XHSN/CUK_ Basophils THSN/CUMM Abnormal L THSWCU _ Other THSN/CU_M GROUP: 2-M 19102 19106 0.3 0.0 0.3 0.0 0.5 0.0 0.2 0.0 19108 19110 0.0 0.0 0.3 0.0 0.3 0.0 0.3 0.0 19113 19120 0.2 0.0 0.0 0.0 0.4 0.0 0.0 0.0 19129 19136 19138 0.6 0.0 0.0 0.0 0.2 0.0 0.0 0.0 0.5 0.0 0.0 0.0 19139 0.0 0.0 0.0 0.0 19147 ..... 19153 ...... 19164 .... 19165 .... 19171 .... MEAN SD N 0.3 0.21 10 0.0 O.(K) 10 0.1 0.14 10 0.0 0.00 10 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-23 SEX: MALE Animal ID Eosinophil THSN/CU MR BasophiLs THSN/CU RM Abnormal L THSN/CU MM Other THSN/CU MR GROUP: 3-M 19101 0.3 0.0 0.1 0.0 19105 0.0 0.0 0.1 0.0 19107 0.5 0.0 0.0 0.0 19112 0.3 0.0 0.1 0.0 19114 0.1 0.0 0.0 0.0 19121 0.4 0.0 0.2 0.0 19123 0.0 0.0 0.1 0.0 19130 0.1 0.0 0.0 0.0 19137 0.2 0.0 0.0 0.0 19146 0.6 0.0 0.0 0.0 19155 ...... 19159 .... 19172 ..... 19173 .... 19174 .... MEAN SD N 0.3 0.21 10 0.0 0.00 10 0.1 0.07 10 0.0 0.00 10 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-24 SEX: MALE Animal ID Eosi_hil THSN/CU .. _phils THSN/CU MR Abnormal L THSN/CU .I, Other THSN/CU .. GROUP: 4-M 19100 0.I 0.0 0.1 0.0 19103 0.0 0.0 0.6 0.0 19104 0.4 0.0 0.4 0.0 19118 0.2 0.0 0.5 0.0 19133 0.9 0.0 0.2 0.0 19141 0.3 0.0 0.2 0.0 19142 0.1 0.0 0.1 0.0 19144 0.4 0.0 0.0 0.0 19148 0.2 0.0 0.0 0.0 19150 0.1 0.0 0.0 0.0 19156 .... 19160 ..... 19162 .... 19163 .... 19166 .... MEAN SD N 0.3 0.26 10 0.0 0.00 10 0.2 0.22 10 0.0 0.00 10 27- JUI_=_ Study INDIVIDUAL Report for Hematology ANIMAL REPORT BY PERIOD : TERMINAL GROUP 418-028:PAGE K-25 SEX: MALE Animal ID Eosinophi I THSN/CU MM Basophi ls THSN/CU RM Abnormal L THSN/CU MR Other THSN/CU MM GROUP: 5-M 19111 0.2 0.0 0.5 0.0 19117 0.4 0.0 0.0 0.0 19124 0.1 0.0 0.0 0.0 19126 0.4 0.0 0.0 0.0 19127 19128 0.1 0.0 0.2 0.0 0.1 0.0 0.0 0.0 19140 0.3 0.0 0.5 0.0 19145 19149 0.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 19152 0.0 0.0 0.0 0.0 19154 ...... 19158 ..... 19168 .... 19169 .... 19170 ..... MEAN SD N 0.2 0.15 10 0.0 0.00 10 0.1 0.21 10 0.0 0.00 10 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-26 SEX: FEMALE Ani_l ID Eosi_il _ils _sN/cu PIN T_SN/CU_t_ Abnormal L THSN/CU_M Other THSt_/CtU_ GROUP: 1-F 19010 .... 19012 19019 0.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 19021 0.3 0.0 0.0 0.0 19023 0.1 0.0 0.1 0.0 19041 0.0 0.0 0.0 0.0 19042 0.0 0.0 0.0 0.0 19044 0.1 0.0 0.0 0.0 19050 0.1 0.0 0.0 0.0 19053 0.0 0.0 0.1 0.0 19065 0.1 0.0 0.1 0.0 19068 .... 19072 .... 19074 .... 19075 ..... MEAN SO N 0.1 0.10 10 0.0 0.00 10 0.0 0.05 10 0.0 O.OO 10 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-27 SEX: FEMALE Animal ID Eosir_il THSN/CU MR Bas_hils THSN/CU MR Abnormal L THSN/CU MM Other THSN/CU MR GROUP: 2-F 19004 0.0 0.0 0.0 0.0 19009 0.1 0.0 0.0 0.0 19016 0.3 0.0 0.0 0.0 19018 0.1 0.0 0.0 0.0 19026 0.0 0.0 0.0 0.0 19036 0.4 0.0 0.0 0.0 19037 0.0 0.0 0.0 0.0 19043 0.1 0.0 0.0 0.0 19047 0.1 0.0 0.0 0.0 19048 0.1 0.0 0.0 0.0 19052 .... 19055 .... 19061 .... 19067 .... 19071 .... MEAN SO N 0.1 0.13 10 0,0 0.00 10 0.0 0.00 10 0.0 0.00 10 27-JUN-2002 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-28 SEX: FEMALE Animal ID Eosinophil THSN/CU MM Basa1_hiLs AbnormaL L Other THSN/CU I11,1 THSN/CU 1,111 THSN/CU 111,1 GROUP: 3-F 19003 0.1 0.0 0.1 0.0 19007 0.0 0.0 0.1 0.0 19008 0.0 0.0 0.1 0.0 19013 0.0 0.0 0.2 0.0 19014 ..... 19015 0.0 0.0 0.0 0.0 19017 0.4 0.0 0.0 0.0 19024 0.2 0.0 0.1 0.0 19029 0.1 0.0 0.0 0.0 19034 0.1 0.0 0.0 O.O 19038 ..... 19056 0.0 0.0 0.0 0.0 19057 .... 19060 .... 19064 .... MEAN SD N 0.1 0.13 10 0.0 0.00 10 0.1 0.07 10 0.0 0.00 10 27-JUN-2(X)2 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-29 SEX: FEMALE Animal ID Eosinophil THSN/CU MM Basophils TMSN/CU MR Abnormal t THSN/CU MM Other THSN/CU MM GROUP: 4-F 19002 .... 19005 0.1 0.0 0.0 0.0 19035 19039 19040 19045 0.0 0.0 0.1 0.0 0.1 O.O 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.O 19046 .... 19051 .... 19054 19058 19062 19063 19066 19069 0.0 0.0 0.2 0.0 0.0 0.0 0.3 0.0 0.0 0.0 .... 0.3 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.1 0.0 19073 ..... MEAN SO N 0.1 0.11 10 0.0 0.00 10 0.1 0.10 10 0.0 0.00 10 27- JUN-2Q02 Study Report for Hematology INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP 418-028:PAGE K-30 SEX: FEMALE Animal ID Eosinophil THSN/CU MR Basophils THSN/CU MM Abnormal L THSN/CU MM Other THSN/CU MM GROUP: 5-F 19001 0.3 0.0 0.0 0.0 19006 0.0 0.0 0.0 0.0 19011 0.0 0.0 0.1 0.0 19020 0.0 0.0 0.0 0.0 19022 0.1 0.0 0.0 0.0 19025 0.0 0.0 0.1 0.0 19027 0.0 0.0 0.0 0.0 19028 0.0 0.0 0.0 0.0 19030 0.0 0.0 0.1 0.0 19031 0.4 0.0 0.0 0.0 19032 ...... 19033 .... 19049 .... 19059 .... 19070 .... MEAN SD N 0.1 0.15 10 0.0 0.00 10 0.0 0.05 10 0.0 0.00 10 27-JUN-2002 Study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL 418-028:PAGE K-31 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE TEST(s) : UNITS: WBC RBC THSN/CU MM MILL/CU NM HGB GRAMS/DL Group: I-M MEAN SD N 15.3 1.63 10 7.51 0.648 10 16.5 1.16 10 HCT MCV MCH % CU MICRONS PICO GRAMS 43.5 3.48 10 57.9 1.89 10 22.0 0.87 10 MCHC PLT % THSN/CU MM MPV CU MICRON 38.0 0.93 10 1179 130.9 10 8.1 0.52 10 Group: 2-M MEAN SD N Group: 3-M MEAN SD N 14.9 3.03 10 7.33 0.396 10 13.8 2.84 10 7.32 0.441 10 15.9 0.42 10 15.7 0.59 10 42.2 1.57 10 42.0 2.22 10 57.7 2.27 10 57.4 1.78 10 21.8 1.13 10 21.5 0.75 10 37.7 1.15 10 1131 183.7 10 37.5 0.79 10 1115 149.2 10 7.9 0.54 10 8.2 0.72 10 Group: 4-M MEAN SD N Group: 5-M MEAN SD N 16.2 3.97 10 6.93* 0.305 10 15.4"* 0.67 10 40.2* 2.29 10 57.9 1.15 10 14.7 3.07 10 6.99* 0.442 10 15.6' 0.81 10 40.7* 1.81 10 58.3 1.37 10 22.2 0.45 10 22.3 0.56 10 38.4 0.80 10 1104 91.4 10 38.3 0.70 10 1148 159.9 10 8.2 0.76 10 8.0 0.59 10 *-Significant Difference from ControL P < .05 LABCAT HE4.43 **-Significant Difference from Control P < .01 27- JUMP2002 Study Report SUMMARY PERIOD: for Hematology REPORT TERMINAL 418-028:PAGE K-32 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE TEST(s) : UNITS: Group: 1-M MEAN SD N PT seconds APTT seconds 13.4 0.24 10 21.0 3.09 10 NRBC Lymphocyte Segmented Bands Monocytes Eosinophi I gasophils COUNTTHSN/CU MM THSN/CU MM THSN/CU MM THSN/EU MM THSN/CU MM THSN/CU MM 0 1! .9 2.2 0.0 0.8 0.3 0.0 0.0 1.72 0.73 0.00 O. 56 0.20 0.00 10 10 10 10 10 10 10 Group: 2-M MEAN SD N Group: 3-M MEAN SD N Group: 4-H MEAN SD N Group: 5-M MEAN SD N 14.2"* 22.1 0 0.27 4.41 0.0 10 10 10 13.6 20.6 0 0.23 2.38 0.0 10 10 10 13.8" 21.2 0 0.40 2.24 0.0 10 10 10 14.0"* 21.3 0 0.47 1 ./+3 0.0 10 10 10 11.0 2.&3 10 10.8 2.02 10 12.1 3.57 10 12.4 2.75 10 2.9 1.18 10 0.0 0.00 10 0.7 0.50 10 0.3 0.21 10 0.0 0.00 10 2.0 1.06 10 0.0 0.03 10 0.6 0.51 10 0.3 0.21 10 0.0 0.00 10 2.8 0.73 10 0.0 0.03 10 0.8 0.67 10 0.3 0.26 10 0.0 0.00 10 1.6 0.78 10 0.0 0,013 10 0.5 0.33 10 0.2 0.15 10 0.0 0.00 10 *-Significant Difference from ControL P < .05 " LABCAT HE4.43 n-Significant Difference from Control P < .01 27- JUM-2002 ! 418-028:PAGE K-33 Study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE TEST(s) : UNITS: AbnormaL L Other THSN/CU NR THSN/CU MM Group: 1-H NEAN SD N O. 1 0.20 10 O. 0 0.00 10 Group: 2-M MEAN SD N 0.1 0.1_ 10 0.0 0.00 10 Group: 3-R MEAN SD N 0.1 0.07 10 0.0 0.00 10 Group: 4-R REAN SD N 0.2 0.22 10 0.0 0.00 10 SEX: HALE Group: 5-R MEAN SO N 0.1 0.21 10 0.0 0.00 10 LABCAT RE4.43 27-JUN-L;_02 418-028:PAGE K-34 Study Report for Hematology SUMMARY PERIOD" REPORT TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE White Group N Blood Tota I Count Mean Std. Dev. DUNNETT'S 't ' DUNflETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm 1-M 10 153.1 15.3 1.63 TREATMENTS 4 2-M 10 149.2 14.9 3.03 0.29 11.9 18.7 11.1 19.5 ERROR 45 3-M 10 137.6 13.8 2.84 1.15 11.9 18.7 11.1 19.5 4-M 10 162.2 16.2 3.97 0.68 11.9 18.7 11.1 19.5 TOTAL 49 5-M 10 146.6 14.7 3.07 0.48 11.9 18.7 11 .I 19.5 F Ratio = 0.90 'F' table values F.CYI : 3.78 F.05 = 2.58 Coeff. Var. % = 20.065 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Red Blood Count I-M 2-11 3-M 4-M 5-M 10 75.13 10 73.26 10 73.19 10 69.30 10 69.89 7.51 7.33 7.32 6.93 6.99 0.6_8 0.396 0.441 0.305 0.442 0.91 0.94 2.83 2.54 7.00 7.00 7.00 7.00 8.03 8.03 8.03* 8.03* 6.87 6.87 6.87 6.87 8.16 8.16 8.16 8.16 TREATMENTS 4 ERROR 45 TOTAL 49 F Ratio = 2.88 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 6.381 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares 32.41 406.23 438.64 2.44 9.54 11.98 Mean Square 8.10 9.03 0.61 0.21 Hemoglobin 1-M 2-M 3-1,1 4-M 5-M 10 165.2 16.5 1.16 TREATMENTS 4 7.40 1.85 10 159.1 15.9 0.42 1.7-I 15.7 17.4 15.4 1716 ERROR 45 26.78 0.60 10 157.3 15.7 0.59 2.29 15.7 17.4 15.4 17.6 10 154.0 15.4 0.67 3.25 15.7 17.4, 15.4 17.6'* TOTAL 49 34.18 10 155.8 15.6 0.81 2.72 15.7 17.4" 15.4 17.6 F Ratio = 3.11 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 4.874 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 *-Significant Difference from Control P < .05 **-Significant Difference from Control P < .01 Error-within groups LABCAT HE4.43 Source-Source of Variation Treatments-between groups 27-JU1_-2002 418-028:PAGE K-35 | Study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL STUDY IO: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DLR4HETT'SPROCEOURE Hematocrit Group N Total Nean 1-M 10 434.6 43.5 2-M 10 422.2 42.2 3-M 10 419.9 42.0 4-M 10 401.7 40.2 5-M 10 406.7 40.7 Std. l)ev. DUNNETT'S 't' 3.48 1.57 2.22 2.29 1.81 1.17 1.39 3.11 2._ DUNNETT's RANGES LO -95%- HI LO -99%.- HI Source Degree Fdm 40.8 40.8 40.8 40.8 46.1 46.1 46.1" 46.1. 40.2 40.2 40.2 40.2 TREATMENTS 4 46.8 ERROR 45 46.8 46.8 46.8 TOTAL 49 F Ratio = Coeff. VaT. % = 5.06 5.676 'F' table values Dunnett's 'T' table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 Mean Corpuscular Volume 1-N 10 579.1 57.9 1.89 2-M 10 576.9 57.7 2.27 0.28 3--R 10 574.1 57.4 1.78 0.64 4-M 10 579.4 57.9 1.15 0.04 5-M 10 582.7 58.3 1.37 0.46 56.0 56.0 56.0 56.0 59.9 59.9 59.9 59.9 55.5 55.5 55.5 55.5 60.3 60.3 60.3 60.3 TREATNENTS 4 ERROR 45 TOTAL 49 F Ratio = 0.34 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. VaT. % = 3.(X)6 Ounnett's 'T' table values P.01 = 3.12 P.05 = 2.51 I-/4 2-M 3-_ 4-M 5-M Mean Corpuscular 10 220.3 10 217.6 10 215.2 10 222.3 10 223.2 22.0 21.8 21.5 22.2 22.3 Hemoglobin O. 87 1.13 0.75 O. 45 0.56 0.76 1.44 O. 57 0.82 21.1 21.1 21.1 21.1 22.9 22.9 22.9 22.9 20.9 20.9 20.9 20.9 23.1 23.1 23.I 23.1 TREATMENTS 4 ERROR 45 TOTAL 49 F Ratio = Coeff. VaT. % = 1.77 3.593 'F' table values Dunnett's 'T' table values F.01 = P.01 = 3.78 3.12 F.05 = P.O5 = 2.58 2.51 SEX: MALE Sum of Squares 68.54 252.15 320.69 Mean Square 17.13 5.60 4.07 136.05 140.12 1.02 3.02 4.40 28.04 32.44 1.10 0.62 *-significant Error-within Difference groups from Control P < .05 LABCAT HE4.43 Source-Source of Variation Treatments-between groups 27- JUN-2002 Study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL 418-028:PAG5 K-36 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE Mean Corpuscular Group N Total Mean Hem. Conc. Std. DUNNETT'S Dev. 't' DUNNETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm 1-M 10 380.4 38.0 O. 93 TREATMENTS 4 2-M 10 377.2 37.7 1.15 0.80 37.0 39.0 36.8 39.3 ERROR 45 3-M 10 374.9 37.5 0.79 1,38 37.0 39.0 36.8 39.3 4-M 10 383.7 38.4 0.80 0.83 37.0 39.0 36.8 39.3 TOTAL 49 5-H 10 383.1 38.3 0.70 0.68 37.0 39.0 36.8 39.3 F Ratio = Coeff. Var. % = 1.81 2.343 'F' table values Ounnett's 'T' table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 Platelets q-H 10 11790 1179 130.9 TREATMENTS 4 2-H 10 11305 1131 163.7 0.74 1015 1343 975 1383 ERROR 45 3-M 10 11154 1115 149.2 0.97 1015 1343 975 1383 4-M 10 11044 1104 91.4 1.14 1015 1343 975 1383 TOTAL 49 5-H 10 11483 1148 159.9 0.47 1015 1343 975 1383 Sum of Squares 5.72 35.64 41.36 Hean Square 1.43 0.79 34523 963815 _R_339 86,,31 21418 F Ratio = Coeff. Vat. % = 0.40 12.888 MEAN P_TELET 'F' table values Dunnett's 'T' table values VOLUME F.01 = P.01 = 3.78 3.12 F.05 = P.D5 = 2.58 2.51 1-M 10 81.2 8.1 0.52 2-M 10 78.6 7.9 0.54 0.92 7.4 8.8 3-M 10 82.3 8.2 0.72 0.39 7.4 8.8 4-M 10 82.1 8.2 0.76 0.32 7.4 8.8 5-M 10 80.4 8.0 0.59 0.28 7.4 8.8 F Ratio = Coeff. Var. % = 0.56 7.823 'F' table Ounnett's values 'T' table values F.01 : P.01 = TREATMENTS 4 7.2 9.0 ERROR 45 7.2 9.0 7.2 9.0 TOTAL 49 7.2 9.0 3.78 F.05 = 2.58 3.12 P.05 = 2.51 0.90 18.03 18.94 0.23 0.40 Error-within groups Source-Source of Variation LABCAT HE4.43 Treatments-between groups 27-JUN'-2002 Study Report for Hematology SUMMARY REPORT PERIOD: TERMINAL 418-028:PAGE K-37 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: NALE Prothrombin Group N Total Mean Std. Dev. DUNNETT'S 't' DUNNETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm I-M 10 133.8 13.4 0.24 TREATMENTS 4 2-N 10 142.3 14.2 0.27 5.63 13.0 13.8" 12.9 13.9"* ERROR 45 3-M 10 136.3 13.6 0.23 1.66 13.0 13.8 12.9 13.9 4-M 10 137.9 13.8 0.40 2.71 13.0 13.8" 12.9 13.9 TOTAL 49 5-M 10 139.5 14.0 0.47 3.77 13.0 13.8" 12.9 13.9_n_ F Ratio = Coeff. Var. % = 9.05 2.448 'F' table values Dunnett's 'T' table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 Sum of Squares 4.13 5.13 9.26 Mean Square 1.03 0.11 Act. Part. Thromboplastin Time 1-M I0 209.8 21.0 3.09 2-R 10 220.5 22.1 4.41 0.83 17.7 24.2 3-M 10 205.7 20.6 2.38 0.32 17.7 24.2 4-M 10 211.7 21.2 2.24 0.15 17.7 24.2 5-M 10 212.8 21.3 1.43 0.23 17.7 24.2 F Ratio = Coeff. Var. % = 0.35 13.609 'F' table values Dunnett's 'T' table values F.01 = P.01 = Nucleated Red Cells 1-M 10 0 0 0.0 2-R 10 0 0 0.0 0.00 3-M 10 0 0 0.0 0.00 0 0 0 0 4-M 10 0 0 0.0 0.00 0 0 5-M 10 0 0 0.0 0.00 0 0 F Ratio = Coeff. Var. % = 0.00 0.000 'F' table values Dunnett's 'T' table values F.01 = P.01 = 17.0 17.0 17.0 17.0 25.0 25.0 25.0 25.0 TREATMENTS 4 ERROR 45 TOTAL 49 3.78 3.12 F.O5 = P.05 = 2.58 2.51 0 0 0 0 3.78 3.12 TREATMENTS 4 0 ERROR 45 0 0 TOTAL 49 0 F.05 = P.05 = 2.58 2.51 11.75 374.92 386.66 2.94 8.33 0.00000 O. O0(X_ 0.00000 O. 00000 O. 00000 *-Significant **-Significant Error-within Difference Difference groups from Control from Control P < .05 P < .01 LABCAT HE4.43 Source-Source of Variation Treatments-between groups 27-JUN-2002 Study Report for Hematology SUMMARY PERIOD: REPORT TERMINAL 418-028:PAGE K-38 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE Lymphocytes Group N Total Mean Std. Dev. DUNNETT'S 1:' DUNNETT*s RANGES LO -95%- HI LO -99%,- HI Source Degree Fdm q-M 10 119.2 11.9 1.72 TREATMENTS 4 2-M 10 109.7 11.0 2.68 0.81 9.0 14.9 8.3 15.6 ERROR 45 3-M 10 108.1 10.8 2.02 0.9& 9.0 14.9 8.3 15.6 4-M 10 120.8 12.1 3.57 0.14 9.0 14.9 8.3 15.6 TOTAL 49 5-M 10 123.6 12.4 2.75 0.37 9.0 14.9 8.3 15.6 F Ratio = 0.70 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 22.619 Dunnet1:'s 'T' table values P.01 = 3.12 P.05 = 2.51 Segmented Neutrophils 1-M 2-M 3-M 4-N 5-M 10 22.1 2.2 O. 73 10 28.8 z.9 1.18 1._ 10 20.2 2.0 1.06 0.46 10 2B.4 2.8 0.73 1.54 10 15.5 1.6 0.78 1.61 1.2 3.2 1.2 3.2 1.2 3.2 1.2 3.2 TREATMENTS 4 0.9 3.5 ERROR 45 0.9 3.5 0.9 3.5 TOTAL 49 0.9 3.5 F Ratio = 3.79 'F' table values F.01 : 3.78 F.05 = 2.58 Coeff. Vat. % : 39.885 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Bands Sum of Squares 19.27 311.29 330.56 Mean Square 4.82 6.92 12.7"7 ]7.87 50.64 3.19 0.84 1-M 10 0.0 0.0 0.00 2-M 10 0.0 0.0 0.00 0.00 0.0 0.0 3-M 10 0.1 0.0 0.03 1.12 0.0 0.0 4-I_ 10 0.1 0.0 0.03 1.12 O.O 0,0 5-M 10 0.0 0.0 0.00 0.00 0.0 0.0 F Ratio = Coeff. Var. % = 0.75 500.000 'F' table values Ounnet1:'s 'T' table values F.01 = P.01 = TREATMENTS 4 0.0 0.0 0.0 0.0 ERROR 45 0,0 0.0 0.0 0,0 TOTAL 49 3.78 3.12 F.05 = P.05 = 2.58 2.51 0.00"12 0.0180 0.0192 0.0003 0.0004 Error-within groups Source-Source of Variation LABCAT HE4.43 Treatmen1:s-between groups 27-JUle-2002 Study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL 418-028:PAGE K-39 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE Monocytes Group N Total Mean Std. Dev. DUI_IETT' S 't' DUNNETT'S RANGES LO-95%- HI LO -99%- HI Source Degree Fdm I-R 2-M 3-M 4-M 5-fl 10 8.4 0.8 0.56 TREATMENTS 4 10 6.9 0.7 0.50 0.64 0.2 1.4 0.1 1.6 ERROR 45 10 5.8 0.6 0.51 1.11 0.2 1.4 0.1 1.6 10 8.0 0.8 0.67 0.17 0.2 1.4 0.1 1.6 TOTAL 49 10 4.5 0.5 0.33 1.6_5 0.2 1.4 0.1 1.6 F Ratio = Coeff. Var. % = 0.93 78.222 'F' table values Dunnett's 'T' table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.58 2.51 1-M 3-M 4-M 5-M Eos inophi I s 10 2.6 0.3 10 3.0 0.3 lO 2.5 0.3 10 2.7 0.3 10 1.8 0.2 0.20' 0.21 O.Zl 0.26 0.15 0.43 O.ll 0.11 0.86 0.0 O.S o.o o.5 0.0 0.5 0.0 0.5 TREATMENTS 4 0.0 0.5 ERROR 45 o.o 0.5 0.0 0.5 TOTAL 49 0.0 0.5 F Ratio = 0.46 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 82.096 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Ba sophi i s q-M 10 0.0 0.0 0.00 2-M 10 0.0 0.0 0.00 0.00 3-_ lO o.o o.o o.oo o.oo 4-R 10 0.0 0.0 0.00 0.00 5-M 10 0.0 0.0 0.00 0.00 TREATMENTS 4 0.0 0.0 0.0 0.0 ERROR 45 o.o o.o o.o o.o 0.0 0.0 0.0 0.0 TOTAL 49 0.0 0.0 0.0 0.0 F Ratio = 0.00 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 0.000 Ounnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares Mea_ Square 1.03 12.43 0.26 0.28 13.46 0.08 0.02 1.93 0.04 2.00 0.00000 O. 00000 0.00000 O. OOOO0 O. 00000 Error-within Source-Source groups of Variation LABCAT HE4.43 Treatments-between groups 27-JUN-2002 418-028:PAGE K-40 I Study Report for Hematology SUMMARY PERIOD: REPORT TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE Abnormal Group N Total Lymphocytes Std. Mean Dev. DUNNETT'S 't' 1-M 10 1.4 0.1 0.20 2-N 10 1 .I 0.1 0.14 0.38 3-M 10 0.6 0.1 0.07 1.01 4-M 10 2.1 0.2 0.22 0.89 5-M 10 1.2 0.1 0.21 0.25 DUNNETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm -.I 0.3 -.1 0.3 -.I 0.3 -.1 0.3 TREATMENTS 4 -.1 0.4 ERROR 45 -.I 0.4 -.I 0.4 TOTAL 49 -.1 0.4 F Ratio = 0.95 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 137.898 Dunnett's 'T' table values P.01 = 3.12 P.O5 = 2.51 Other 1-N 10 0.0 0.0 0.00 TREATMENTS 4 2-R 10 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 ERROR 45 3-M 10 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 4-M 10 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 TOTAL 49 5-M 10 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 F Ratio = 0.00 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 0.000 Ounnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares 0.12 1.40 1.52 Mean Square 0.03 0.03 0.00000 0.00000 0.00000 O. 00000 0. OOO00 Error-within Source-Source groups of Variation LABCAT HE4.43 Treatments-between groups 27-JUN-2002 Study Report for Hematology SUMMARY PERIOD: REPORT TERMINAL 418-028:PAGE K-41 STUDY IO: ARGUS 418-028 STUDY NO: 060--069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: FEMALE TEST(s ) : UNITS: WBC RBC THSN/CU MR MILL/CU ttM HGB GRAMS/DL GPoup: 1-F MEAN SD N 9.0 3.09 10 6.51 0.461 10 15.5 0.95 10 HCT MCV NCH % CU MICRONS PICO GRAMS MCHC PLT % THSN/CU MN MPV CU MICROtl 40.5 2.93 10 62.2 1.19 10 23.9 0.55 10 38.4 0.76 10 1419 153.0 10 8.3 0.49 10 Group: 2-F MEAN 5D N 13.8" 5.73 10 6.22 0.423 10 15.0 0.77 10 39.3 2.72 10 63.3 2.01 10 24.2 0.76 10 38.3 0.94 10 1240 208.1 10 8.0 0.84 10 Group: 3-F IIEAN SD N 11.1 2.98 10 6.40 0.554 10 15.2 0.91 10 39.5 3.24 10 61.8 1.50 10 23.9 0.88 10 38.7 1.29 10 1274 226.3 10 8.3 0.82 10 Group: 4-F MEAN SD N 10.8 3.15 10 6.39 0.532 10 15.2 0.90 10 40.1 3.02 10 62.9 2.04 10 23.8 1.25 10 37.8 1.22 10 1305 207.2 10 8.0 0.63 10 6roup: 5-F MEAN SD N 8.9 3.48 10 6.42 0.538 10 15.3 0.77 10 40.4 3.15 10 63.0 1.60 10 23.8 1.05 10 37.8 1.37 10 1355 176.8 10 7.7 0.74 10 *-Significant Difference from ControL P < .05 LABCAT HE4.43 27-JUW-2002 study Report for Hematology SUMMARY PERIOD- REPORT TERMINAL 418-028:PAGE K-42 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOgED BY DUNNETTS PROCEDURE SEX: FEMALE TEST(s): UNITS: Group: 1-F MEAN SD N PT seconds APTT seconds 13.2 0.41 10 21.7 2.36 10 NRBC Lymphocyte Segmented Bands Ronocytes EosinophiL BasophiLs COUNTTHSN/CU MR THSN/CU NN THSN/CU NN THSN/CU MM THSN/CU MR THSN/CU NN 0 6.6 2.2 0.0 0.1 0.1 0.0 0.0 2.44 0.90 0.00 0.15 0.10 0.00 10 10 10 10 10 10 10 Group: 2-F MEAN SD N Group: 3-F MEAN SD N 13.4 0.29 10 13.2 0.37 10 23.9 3.19 10 ?3.5 2.35 10 0 9.8* 3.6 0.0 0.3 0.1 0.0 0.0 3.44 3.69 0.00 0.49 0.13 0.00 10 10 10 10 10 10 10 0 8.4 2.4 0.0 0.2 0.1 0.0 0.0 2.60 0.93 0.00 0.13 0.13 0.00 10 10 10 10 10 10 10 Group: 4-F REAN SO N Group: 5-F MEAN SD N 13.5 0.42 10 13.8 1.12 10 25.3 2.45 10 25.2 3.81 10 0 8.2 2.4 0.0 0.2 0.1 0.0 0.0 2.52 0.96 0.09 0.11 0.11 0.00 10 10 10 10 10 10 10 0 6.0 2.6 0.0 0.2 0.1 0.0 0.6 2.77 0.82 0.00 0.14 0.15 0.00 10 10 10 10 10 10 10 *-Significant Difference LABCAT HE4.43 from Control P < .05 27-JUN-2002 STUDY ID: ARGUS 418-028 STUDY NO: 060-009 Study Report for Hematology SUMMARY PERIOD: REPORT T_RMINAL 418-028:PAGE K-43 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE TEST(s) : UNITS: Abnornml L Other THSN/CU I_ THSN/CU MM Group: 1-F MEAN SD N 0.0 0.05 10 0.0 0.00 10 SEX: FEMALE Group: 2-F MEAN SD N 0.0 0.00 10 0.0 0.00 10 Group: 3-F MEAN SD N O. 1 0.07 10 0.0 0.00 10 Group: 4-F MEAN SD N 0.1 0.10 10 0.0 0.00 10 Group; 5-F MEAN SD N 0.0 0.05 10 0.0 0.00 10 LABCAT HE4.43 27- JUN-2O_ Study Report for Hematology SUMMARY REPORT PERIOD: TERMINAL 418-028:PAGE K-44 STUDY ID: ARGUS418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE White Group N Blood Total Count Mean Std. Dev. DUNNETT'S ' t' q-F 10 89.9 9.0 3.09 2-F 10 137.8 13.8 5.73 2.80 3-F 10 110.7 11.1 2.98 t .22 4-F 10 108.4 10.8 3.15 1.08 5-F 10 88.9 8.9 3.48 0.06 DUNNETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm 4.7 13.3. 4.7 13.3 4.7 13.3 4.7 13.3 TREATMENTS 4 3.6 14.3 ERROR 45 3.6 14.3 3.6 14.3 TOTAL 49 3.6 14.3 F Ratio = Coeff. Var. % = 2.70 35.721 'F' table Ounnett's values 'T' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 Red Blood Count 1-F 10 65.06 6.51 0.461 2-F 10 62.15 6.22 0.423 1.29 5.94 7.07 3-F 10 63.96 6.40 0.554 0.49 5.94 7.07 4-F 10 63.87 6.39 0.532 0.53 5.94 7.07 5-F 10 64.23 6.42 0.538 0.37 5.94 7.07 F Ratio = Coeff. Var. % = 0.44 7.896 'F' table Dunnett's values 'T' table values F.01 = P.01 = TREATNENTS 4 5.80 7.21 5.80 7.21 ERROR 45 5.80 7.21 TOTAL 49 5.80 7.21 3.78 F.05 = 2.58 3.12 P.05 = 2.51 Hemoglobin 1-F 10 155.4 15.5 2-F 10 150.4 15.0 3-F 10 152.4 15.2 4-F 10 151.6 15.2 5-F 10 152.6 15.3 0.95 0.77 0.91 0.90 0.77 1.29 0.78 0.98 0.72 14.6 14.6 14.6 14.6 16.5 16.5 16.5 16.5 14.3 14.3 14.3 14.3 16:7 16.7 16.7 16.7 TREATNENTS 4 ERROR 45 TOTAL 49 F Ratio = 0.46 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 5.669 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 SEX: FEMALE Sum of Squares 158.42 659.12 817.54 Mean Square 39.61 14.65 0.45 0.11 11.44 0.25 11.89 1.36 0.34 33.62 0.75 34.98 *-Signifir_nt Error-within Difference groups from Control P < .05 LABCAT HE4.43 Source-Source of Variation Treatments-between groups 27-JUN-2(X]2 418-028:PAGE K-45 Ii study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE Hematocrit Group 1-F 2-F 3-F 4-F 5-F N Total I0 404.9 10 393.4 10 395.1 10 401.4 10 404.1 Mean 40.5 39.3 39.5 40.1 40.4 Std. Dev. DUNNETT'S 't' 2.93 2.72 3.24 3.02 3.15 0.85 0.73 0.26 0.06 DUNNETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm 37.1 37.1 37.1 37.1 43.9 43.9 43.9 43.9 36.3 36.3 36.3 36.3 44.7 44.7 44.7 44.7 TREATMENTS 4 ERROR 65 TOTAL 49 F Ratio = Coeff. Var. % = 0.30 7.548 'F' table Dunnett's values 'T' table values F.01 = P.01 = Mean Corpuscular Volt, me 1-F 10 622.3 62.2 1.19 2-F 10 633.3 63.3 2.01 1.45 60.3 64.1 3-F 10 618.2 61.8 1.50 0.54 60.3 64.1 4-F 10 629.1 62.9 2.04 0.90 60.3 64.1 5-F 10 629.7 63.0 1.60 0.97 60.3 64.1 3.78 F.05 = 2.58 3.12 P.05 = 2.51 59.9 59.9 59.9 59.9 64.6 64.6 64.6 64.6 TREATMENTS 4 ERROR 45 TOTAL 49 F Ratio = 1.30 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 2.709 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Mean Corpuscular Hemoglobin 1-F 10 239.0 23.9 0.55 2-F 10 242.4 24.2 0.76 0.82 _--F 10 2,38.9 23.9 o.88 o.o2 4-E 10 237.9 23.8 1.25 0.26 5-F 10 238.3 23.8 1.05 0.17 22.9 22.9 22.9 22.9 24.9 24.9 24.9 24.9 22.6 22.6 22.6 22.6 25.2 25.2 25.2 25.2 TREATItE, NTS 4 ERROR 45 TOTAL 49 F Ratio = 0.37 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 3.886 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 SEX: FEMALE Sum of Squares 11.01 409.80 420.81 Mean Square 2.75 9.11 14.98 129.67 144.64 3.74 2.88 1.28 0.32 38.92 0.86 40.21 Error-within groups Source-Source of Variation LABCAT HE4.43 Treatments-between group 27- JUN-2002 Study Report for Hematology SUMMARY PERIOD: REPORT TERMINAL 418-028:PAGE K-46 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DLINNETT'S PROCEDURE SEX: FEMALE Mean Corpuscular Group 1- F 2-F 3-F 4-F 5-F N Total 10 384.2 10 382.9 10 386.6 10 378.3 10 378.4 Mean 38.4 38.3 38.7 37.8 37.8 Hem. Cone. Std. DUNNETT'S Dev. 't' 0.76 O. 94 0.26 1.29 0.47 1.22 1.16 1.37 1.14 DUNNETT'S RANGES LO -95%- HI LO -99Z- HI 37. I 39.7 37.1 39.7 37.1 39.7 37.1 39.7 36.8 40.0 36.8 40.0 36.8 40.0 36.8 40.0 Source Degree Fdm TREATMENTS 4 ERROR 45 TOTAL 49 F Ratio = 1.03 'F' table values F.01 = 3.78 F.05 = 2.58 coeff. Var. % = 2.978 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Platelets 1-F 10 14189 1419 153.0 2-F 10 12396 1240 208.1 2.05 1199 1639 3-F 10 12735 1274 226.3 1.66 1199 1639 4-F 10 13048 1305 207.2 1.30 1199 1639 5-F 10 13554 1355 176.8 0.72 1199 1639 F Ratio = Coeff. Var. % = 1.29 14.868 'F' table Dunnett's values 'T' table values F.01 = P.01 = MEAN PLATELET VOLUME 1145 1145 1145 1145 1692 1692 1692 1692 TREATMENTS 4 ERROR 45 TOTAL 49 3.78 F.05 = 2.58 3.12 P.05 = 2.51 1-F 10 83.4 8.3 0.49 TREATMENTS 4 2-F 10 80.0 8.0 0.84 1.06 7.5 9.1 7.3 9.3 ERROR 45 3-F 10 82.7 8.3 0.82 0.22 7.5 9.1 7.3 9.3 4-F 10 80.3 8.0 0.63 0.97 7.5 9.1 7.3 9.3 TOTAL 49 5-F 10 76.5 7.7 0.74 2.15 7.5 9.1 7.3 9.3 F Ratio = 1.44 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % : 8.894 Ounnett's 'T' table values P.01 = 5.12 P.05 = 2.51 Sum of Squares 5.34 58.25 63.60 198797 1729216 1928012 2.95 23.11 26.06 Mean Square 1.34 1.29 49699 38427 0.74 0.51 Error-within groups Source-Source of Variation LABCAT HE4.43 Treatments-between groups 27- JUN-2002 study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL 418-028:PAGE K-47 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: FEMALE Prothrombin Group N Total Mean Std. Dev. DUNNETT'S 't' DUNNETT'S RANGES LO -95%- HI LO-99%- HI Source Degree Fdm 1-F 10 131.8 13.2 O.41 TREATMENTS 4 2-F 10 133.5 13.4 0.29 0.63 12.5 13.9 12.3 14.0 ERROR 45 3-F 10 131.7 13.2 0.37 0.04 12.5 13.9 12.3 14.0 4-F 10 134.8 13.5 0.42 1.11 12.5 13.9 12.3 14.0 TOTAL 49 5-F 10 137.5 13.8 1.12 2.11 12.5 13.9 12.3 14.0 F Ratio = 1.58 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. VaT. % : 4.521 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Act. Part. Thromboplastin Time 1-F 10 217.3 21.7 2.36 TREATMENTS 4 2-F 10 239.4 25.9 3.19 1.71 18.5 25.0 17.7 25.8 ERROR /,5 3-F 10 234.5 23.5 2.35 1.33 18.5 25.0 17.7 25.8 4-F 10 252.5 25.3 2.45 2.72 18.5 25.0* 17.7 25.8 TOTAL 49 5-F 10 252.1 25.2 3.81 2.89 18.5 25.0* 17.7 25.8 F Ratio = 2.53 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. VaT. % : 12.081 Ounnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Nucleated Red Cells Sum of Squares 2.32 16.48 18.80 Mean Square O. 58 0.37 84.50 375.66 460.17 21.13 8.35 1-F 2-F 3-F 4mE 5-F 10 0 0 0.0 10 0 0 0.0 0.00 10 0 0 0.0 0.00 10 0 0 0.0 0.00 10 2 0 0.6 1.58 TREATMENTS 4 0.32 0.08 0 0 0 0 ERROR 45 3.60 0.08 0 0 0 0 0 0 0 0 TOTAL 49 3.92 0 0 0 0 F Ratio = 1.00 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % : 707.107 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 *-Significant Difference from Control Error-within groups P < .05 LABCAT HE4.43 Source-Source of Variation Treatments-between groups 27-JUN--2002 418-028:PAGE K-48 Study Report for Hematology S_Y PERIOD- _PORT TERMINAL STUDY ID: ARGUS 418028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETTS PROCEDURE SEX: FEMALE Lymphocytes Group N TotaL Mean Std. Dev. DUNNETT'S 't' DUNNETTS RANGES LO -95%- HI LO -99%- HI Source Degree Fdm 1-F 10 65.6 6.6 2.44 TREATMENTS 4 2-F 10 98.0 9.8 3.44 2.61 3.4 9.7* 2.7 10.6 ERROR 45 3-F 10 83.8 8.4 2.60 1.46 3.4 9.7 2.7 10.4 4-F 10 81.8 8.2 2.52 1.30 3.4 9.7 2.7 10.4 TOTAL 49 5-F 10 60.3 6.0 2.77 0.43 3.4 9.7 2.7 10.4 F Ratio = 2.96 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 35.677 Ounnett's 'T' tabLe values P.01 = 3.12 P.05 = 2.51 Segmented Neutrophils I-F 10 21.9 2.2 0.90 2-F 10 36.0 3.6 3.69 1.72 3-F 10 23.8 2.4 0.93 0.23 6-F 10 23.5 2.4 0.96 0.19 5-F 10 25.9 2.6 0.82 0.49 0.1 4.3 0.1 4.3 0.1 4.3 0.1 4.3 TREATHENTS 4 -.4 4.8 ERROR 45 -.4 4.8 -.6 6.8 -.4 4.8 TOTAL &9 F Ratio = 0.95 'F' table vaLues F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 70.025 9unnett_s 'T' table values P.01 = 3.12 P.05 -- 2.51 Bands 1-V 2-F 3-F 4-F 5-F 10 0.0 0.0 0.00 10 0.0 0.0 0.00 0.00 10 0.0 0.0 0.00 0.00 10 0.3 0.0 0.09 1.58 10 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 TREATMENTS 4 -.1 0.1 ERROR 45 -.1 0.1 -.I 0.1 TOTAL 49 -.1 0.1 F Ratio = Coeff. Vat. % = 1.00 707.107 'F' table Dum_ett's values 'T' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 Sum of Squares 91.51 347.60 639.11 12.77 151.70 164.67 0.0072 0.0810 0.0882 Mean Square 22.88 7.72 3.19 3.37 0.0018 0.0018 *-Significant ErPoP-within Difference groups from Control P < .05 LABCAT HE4.43 Source-Source of Variation Tl'eataents-between gPoups 27-JUN-2002 Study Report for Hematology SUMMARY REPORT PERIOD: TERMINAL 418-028:PAGE K-49 STUDY ID: AR6US 4.18-028 STUDY NO: 060-069 ANALYSIS OF VARXANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: FEPIALE Monocytes Group N Total Hean Std. Dev. DUNNETT'S 't' DUNNETT'S RANGES LO-95%- HI tO -99%- HI Source Degree Fda I-F 10 1.3 0.1 0.15 TREATMENTS 4 2-F 10 2.7 0.3 0.69 1.26 -.1 0.4 -.2 0.5 ERROR 45 3-F 10 1.7 0.2 0.13 0.36 -.1 0.4 -.2 0.5 4-F 10 1.9 0.2 0.11 0.54 -.1 0.4 -.2 0.5 TOTAL 49 5-F 10 1.7 0.2 0.14 0.36 -.1 0.4 -.2 0.5 F Ratio = 0.43 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. 7. = 133.9A2 Ounnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares 0.11 2.79 2.90 Mean Square 0.03 0.06 Eosinophils q-F 10 0.9 0.1 2-F 10 1.2 0.1 3-F 10 0.9 0.1 4-F 10 0.7 0.1 5-F 10 0.8 0.1 0.10 0.13 0.54 0.13 0.00 0.11 0.36 0.15 0.18 0.0 0.2 0.0 0.2 0.0 0.2 0.0 0.2 F Ratio = Coeff. Var. g = 0.23 137.686 'F' table Dunnett's values 'T' table values F.oq = P.01 = TREATMENTS 4 -.1 0.3 ERROR 45 -.1 0.3 -.1 0.3 TOTAL 49 -.1 0.3 3.78 F.05 = 2.58 3.12 P.05 = 2.51 0.014 0.691 0.705 0.004 0.015 Bas ophi is 1-F 10 0.0 0.0 0.00 2-F 10 0.0 0.0 0.00 0.00 3-F 10 0.0 0.0 0.00 0.00 6-F 10 0.0 0.0 0.00 0.00 5-F 10 0.0 0.0 0.00 0.00 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 F Ratio = Coeff. Var. % = 0.00 O.O(X) ' F' 1:able values Dunnett's 'T' table values F.01 = P.01 = TREATHENTS 4 0.0 0.0 ERROR45 0.0 0.0 0.0 0.0 TOTAL 49 0.0 0.0 3.78 f.05 = 2.58 3.12 P.05 = 2.51 0.0(3000 O. 00000 0.00000 0.00000 0.00000 Er_r_ithin groups Source-Source of Variation LABCAT HE4.43 TreatRnt_t_ groups _-J_2002 418-028:PAGE K-50 11 Study Report for Hematology SUMMARY REPORT PERIOD : TERMINAL STUDY ID: ARGUS &18-028 STUDY NO: 060-069 ANALYSIS OF VARZANCE FOLLOWEDBY DUNNETF'S PROCEDURE SEX: FEMALE Abnormal Lymphocyt Group N Tota I Hean es Std. Dev. DUNNETT'S Jt' DUNNETT'S RANGES LO -95%- HI LO -99_- HI Source Degree Fdm 1-F 10 0.3 0.0 0.05 TREATIIENTS 4 2-F 10 0.0 0.0 0.00 1.09 0.0 0.1 -.1 0.1 ERROR 45 3-F 10 0.6 0.1 0.07 1.09 0.0 0.1 -.1 0.1 4-F 10 0.5 0.1 0.10 0.73 0.0 0.1 -.1 0.1 TOTAL 49 5-F 10 0.3 0.0 0.05 0.00 0.0 0.1 -.1 0.1 F Ratio = 1.39 'F' tabLe vaLues F.01 = 3.78 F.05 = 2.58 Coeff. VaT. % = 181.306 Dunnett's 'T' tabLe vaLues P.(Yl = 3.12 P.05 = 2.51 Other I-F 10 2-F 10 _F lo 4-F 10 5-F 10 F Ratio = Coeff. VaT. % : 0.0 0.0 0.00 0.0 0.0 0.00 0.00 o.o o.o o.oo o.oo 0.0 0.0 0.00 0.00 0.0 0.0 0.00 0.00 0.0 0.0 o.o 0.o 0.0 0.0 0.0 0.0 0.00 0.000 'F' table Dunnett's values 'T' table values F.01 = P.01 = TREATMENTS & 0.0 0.0 0.o o.o ERROR 45 0.0 0.0 TOTAL 49 0.0 0.0 3.78 F.05 = 2.58 3.12 P.05 = 2.51 Sum of Squares 0.021 0.171 0.192 O. 00000 0.000(30 0.00000 Mean Square 0.005 0.004 0.00000 0.00000 Error-within groups Source-Source of Variation LABCAT HE4.43 Treatments-betueen groups 27-Jl,nt-2002 418-028:PAGE K-5I Study Report for Hematology WHITR DIFFERRNTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID GROUP: 1-M 19109 19115 19116 Nucleated Red CeLls Lyaphocytes Segmented Neutrophi ls Bands Monocytes Eosinophi ts Basophi ls Abnormal Other LylphocyCes WBC Nucleated Red Cells Lywphocytes Segmented Neutrophi Ls Bands I_cytes Eosinophi ls Basophi ls AbnormmL Lymphocytes Other WBC NucLeated Red CelLs Ly_o_ocytes Segmented Neutrophi ls Bands Monocytes Eosinophi ts Basophi Ls Abnormal Lymphocytes OCher WBC TERMINAL CNT ABS 0 72 9.7 15 2.0 0 O. 0 8 1.1 5 O. 7 0 O. 0 0 0.0 0 0.0 13.5 0 72 11.4 17 2.7 0 0.0 10 1.6 1 O. 2 0 0.0 0 0.0 0 O. 0 15.9 0 77 11.6 20 3.0 0 O. 0 2 0.3 1 0.2 0 0.0 0 0.0 0 O. 0 15.1 SEX: MALE LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-52 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS /+18-028 STUDY NO: 060-069 Animal ID 19119 19122 19125 GROUP: 1-M NucLeated Red CeLLs Lylmphocytes Segmented Neutrophi Ls Bands Honocytes Eosinophi ls Basophi Ls AbnormaL Lymphocytes Other WBC NucLeated Red CeLLs Lymphocyt es Segmented Neut rophi Ls Bands H_ytes Eosinophi Ls Em_i Ls AbnormaL Lyaphocyt es Other WBC NucLeated Red CeLLs Lymphoc)rtes Segmented Neutrophi Ls Bands l_ocyt es Eosinophi Ls Basophi Ls Abnorma I Lymphocytes Other UBC TERMINAL CNT ABS 0 72 9. I 17 2.2 0 0.0 10 1.3 1 O. 1 0 0.0 0 O. 0 0 0.0 12.7 0 86 14.1 9 1.5 0 0.0 1 O. 2 1 O. 2 0 O. 0 3 0.5 0 0.0 16.4 0 81 14.3 8 1.4 0 O. 0 6 1.1 3 0.5 0 0.0 2 O. 4 0 0.0 17.6 SEX: MALE LABCAT HE4. &:3 27- JUN-2002 418-028:PAGE K-53 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ID GROUP:1-M 19131 19132 19134 Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands Honocytes Eosinophi Ls bsophi Ls Abnormal Lymp_ocytes Other WBC Nucleated Red Cells Lymphoc)rtes Segmented Neutrophi Ls Bands leanocy_es Eosinophi Ls BamophiLs id3noru I Lylq)hocytes Other WBC Nucleated Red Cells Lylq)hocytes SegmentedNeutrophi ls Bands le_ocytes E_il_'li Ls Sasophi Ls Abnormal L)nmphocyl:es Other gBC TERRINAL CNT ABS 0 74 13.1 15 2.7 0 O.0 9 1.6 1 0.2 0 O.0 1 0.2 0 0.0 17.7 0 85 12.7 10 1.5 0 0.0 3 0.4 0 0.0 0 O.0 2 0.3 0 O.0 14.9 0 73 11.0 23 3.5 0 O.0 3 O.5 1 O.2 0 O.0 0 0.0 0 0.0 15.0 SEX: MALE LABCATHE4.63 27- JUN-2002 418-028:PAGE K-54 Study Report for Hematology WHITE DII_3'I_RENTTAL DATA STUDY ID: ARGUS 418-028 STUOY NO: 060-069 Animal ID GROUP: 1-H TERfllNAL CNT ABS 19135 19143 Nucleated Red Cells Ly_ohocytes Segmented Neutr_i Ls Bands Monocy_ es Eosinophi Ls bsophi Ls Abnormal Lywq)hocytes Other WBC Nucleated Red CeLls Lylq)hocyt es Segmented Neutrophi Ls Bands Honocytes Eosinophi Ls Basophi Ls Abnormal Lymphocytes Other WBC 0 85 12.2 11 1.6 0 0.0 2 O. 3 2 0.3 0 0.0 0 0.0 0 0.0 14.3 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- -- 19151 Nucleated Red CelLs L)quphocytes Segmented Neutrophi ls Bands l$onocytes Eosir_p_li ls Basophi ts AbnormaL Ly_cyCes Other 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: MALE (--) - Data Unavailable _BCAT HE4.43 27-JUN-_ 418-028:PAGE K-55 i Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY %D: AR6US 41_028 STUDY NO: 060-069 Animal ID GROUP: 1-M TERMZNAL CNT ABS SEX: I_LE 19157 NucLeated Red CeLLs Lymphocytes segmented Neutrophi Ls Bands Ronocytes Eosinophi Ls bsophi ts Abnormal Lyephocyl:es Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 19161 19167 NucLeated Red CeLLs Lytphocytes Segmented Neutrophi Ls Bands Honocytes Eosi nophi Ls Basophi ts Abnormal Lymphocyl:es Other WBC NucLeated Red CeLLs Lymphocytes Segmented Neutr_hi ls Bands Honocytes Eosi nophi Ls Basophi ls AbnormaL LymphocyCes Other ttBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- (--) - Data Unavai LabLe LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-56 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ID 19102 19106 19108 GROUP:2-M NucLeated Red CeLLs LymphocyCes SegmencedNeutrophi ls Bands Monocytes Eosinophi Ls Basophi ts AbnormaLLymphocytes Other WBC NucLeated Red CeLLs Ly_hocyCes SegmentedNeutrophi Ls Bands Honocytes Eosinophi Ls Basophi Ls Abnormal Lya_hocyCes Other WBC NucLeated Red CeLLs Lywphocytes Segmented NeutrophiLs Bands Monocytes Eosinophi Ls BasophiLs Abnormal Lymphocy_es Other WBC TERHINAL CNT ABS 0 60 6.5 25 2.7 0 0.0 9 1.0 3 0.3 0 0.0 3 0.3 0 0.0 10.9 0 71 12.1 17 2.9 0 0.0 8 1.4 3 0.5 0 0.0 1 0.2 0 0.0 17.0 0 86 10. 9 11 1.4 0 0.0 1 O.1 0 0.0 0 0.0 2 0.3 0 0.0 12.7 SEX: HALE LABCATHE4.43 27-JUN-20(}2 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-57 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Anima IID GROUP: 2-M TERMINAL CNT ABS SEX: MALE 19110 19113 19120 NucLeated Red CeLls Lymphocytes Segmented Neutrophi Ls Bands Monocyt es Eosinophi ls BasophJ Ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocyt es Segmented Neutrophi Ls Bands l_ocytes Eosinophi Ls Basophi Ls Abnormal Lymphocytes Other _BC Nucleated Red Cells Lymphocyt es Segmented Neutrophj ls Bands Monocytes Eosinophi Ls Basophi Ls Abnormal Lymphocytes Other WBC 0 76 10.0 15 2.0 0 O. 0 5 O. 7 2 0.3 0 O. 0 2 0.3 0 0.0 13.I 0 71 11.1 23 3.6 0 0.0 5 O. 8 I 0.2 0 O. 0 0 0.0 0 O. 0 15.7 0 73 13.4 17 3.1 0 O. 0 8 I.5 2 0.4 0 O. 0 0 0.0 0 O. 0 18.4 LABCAT HE4. A5 27- JUN-2002 418-028:PAGE K-58 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: AR6US&18-028 STUDYNO: 060-069 Animal ID GROUP:2-N 19129 NucLeated Red CeLLs Lymphocytes Segmented Neutrophi Ls Bands ttormcytes Eosirmphi Ls Basophi Ls - Abnormal L)qephocyets Other UBC 19136 NucLeated Red CeLLs Lymphocytes SegmentedNeutrophi ts Bands Monocytes Eosinophi Ls ksophi Ls Abnormal Lymphocytes Other I_c 19138 NucLeated Red CeLLs Lyephocytes SegmentedNeutraphi Ls Monocytes Eosinophi Ls bsophi Ls Abnormal Lywphocytes Other UBC TERNINAL CNT ABS 0 64 9.4 27 4.0 0 O.0 5 O.7 4 0.6 0 O.0 0 0.0 0 0.0 14.7 0 82 8.6 13 1.& O 0.0 3 0.:3 Z 0.2 0 O.0 0 0.0 0 0.0 10.5 0 66 11.4 30 5.2 0 0.0 1 O.2 3 0.5 0 0.0 0 0.0 0 O.0 17.2 SEX: NALE LABCATHE4.43 27-JUN-2002 418-028:PAGE K-59 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 GROUP:2-M Animal ID TERNINAL CNT ABS 19139 19147 19153 NucleatedRed CelLs kya_hocytes SegmentedNeutrophi ls Bands Honocytes Eosinop_i ls Basophi Ls AbnormaLLymphocytes Other WBC NucLeated Red CeLLs Lymphocyl:es SegmentedNeutrophiLs Bands ttonocytes Eosinophi ts BasophiLs AbnormaLL)_ocyt es Other gBC NucLeated Red CeLLs L)m_nocytes Segmented NeutrophiLs Bamls Monocytes Eosinophi Ls Basophi Ls AbnormaL Lymphocyl:es Other gBC 0 86 16.3 13 2.5 0 O.0 1 0.2 0 0.0 0 0.0 0 0.0 0 0.0 19.0 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: MALE (--) - Data Unavailable LABCATHE4.43 27-JU1_2002 418-028:PAGE K-60 im Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: AR6US 418-028 STUDY NO: 060-069 Animal ID 19164 19165 19171 GROUP: 2-M NucLeated Red Cells Lymphocytes Segmented Neutrophi Ls Bands Nonocytes Eosinophi ls Basophi Ls Abnormal Lymphocyt es Other WBC Nucleated Red Cells LymphocyCes Segmented N_tr_i ls Bands Ronocytes Eosi nophi ls Basophi ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lyaphocytes Segmented Neutrophi ls Bands Nonocytes Eosinophi ls Basophi ls Abnormal Lymphocyl:es Other WBC TERRIHAL CNT ABS 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- '0 -- 0 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- -- SEX: NALE (--) - Data Unavailable LABCAT HE4.43 27- JUN-2002 418-028:PAGE K-61 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: AR6US418-028 STUDYNO: 060-069 Animal ZD 19101 19105 19107 GROUP:3-R NucLeated Red CeLLs LyBphocytes SegmentedNeutrophi Ls Bands Monocy_es Eosinophi Ls Basophi Ls AbnormaLLymphocytes OCher WBC Nucleated Red Cells L)_phoc)rtes SegmenCedNeutrophi Ls Bands Ronocytes Eosinophi Ls Basophi Ls Abnormal L)mphocytes other _BC Nucleated Red Cells tymphocy_es Segmented_r_phi Ls Bands Monocytes Eosinophi Ls Basophits Abnormal Lyq0hocytes Other WBC TERRZNAL CNT ABS 0 77 10.1 16 2.1 0 O.0 4 O.5 2 0.3 0 0.0 I O.I 0 0.0 13.1 0 80 8.8 17 1.9 0 0.0 2 0.2 0 0.0 0 0.0 1 0.1 0 0.0 11.0 0 78 9.8 5 0.6 1 0.1 12 1.5 4 O.5 0 0.0 0 0.0 0 0.0 12.5 SEX: RALE LABCATHE4._3 27-JUN-2002 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-62 STUDY /D: AR6US 418-028 STUOY NO: 060-069 AnimaL ID 6ROUP: 3-ti TERMINAL CNT AB5 SEX: I_ALE 19112 19114 19121 NucLeated Red CeLLs Lymphocytes Segmented Neutrophi ls Bands Monocytes Eosi nophi Ls Sasophi ls Abnormal Lywhocytes Other WBC Nucleated Red Cells L)_ohoc)rt es Segmented Neutrophi Ls Bands Ronocyt es Eosinophi Ls Basophi ls Abnormal Ly_hocyt es Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi ls Bands Monocytes Eosinophi ls Basophi ls Abnormal Lymphocytes Other WBC 0 83 11.0 10 1.3 0 O. 0 4 0.5 2 0.3 0 O. 0 1 0.1 0 0.0 13.3 0 70 7.8 24 2.7 0 0.0 5 O. 6 1 O. 1 0 0.0 0 0.0 0 0.0 11.2 0 71 14.8 19 4.0 0 0.0 7 1.5 2 0.4 0 0.0 1 0.2 0 0.0 20.9 LABCAT HE4.43 27-JUN-20G2 418-028:PAGE K-63 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 GROUP:_-R SEX: MALE Animal IO TERMINAL CNT ABS 19123 19130 19137 NucLeated Red CeLls Lymphocytes SegmentedNeutral Ls Bands Ronocytes Eosinophi Ls bsophi ls Abnormal Lynphocyt:es Other _C Nucleated Red CeLLs L>_4_hocyet s SegmentedNeutrophi Ls Bands Monocytes Eosinophi ls BasophiLs Abnormal Lyaq:d_cytes Other WBC NucLeated Red CeLLs Lymphocytes _ted Bands Neutrophi Ls Ronocytes Eosinophi Ls Basophi Ls Abnormal Lyaphocytes Other WBC 0 74 TI0.4 24 3.4 0 0.0 1 O.1 0 0.0 0 0.0 1 0.1 0 0.0 14.1 0 84 11.7 12 1.7 0 O.0 3 0.4 1 O.1 0 O.0 0 0.0 0 O.0 13.9 0 87 10.6 9 1.1 0 0.0 2 O.2 2 0.2 0 O.0 0 0.0 0 0.0 12.2 LABCATHE4.43 27-JUN-2002 418-028:PAGE K-64 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ID GROUP:_ TERMINAL CNT ABS 19146 19155 19159 Nucleated Red Cells tya_hocytes Segmented Neutroph_ ks Bands l_ocytes Eosinophi Ls BasophiLs Abnormal Lym_0hocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi ls Bands _onocytes Eosinophi Ls Ba=ophiLs AbnormaL tyRohocytes Other _BC NucLeated Red CeLLs Lymphocytes Segmented Neutrophits Bands l_nocytes Eosinophi ls Base_W_lis AbnormaL Lymphocytes Other WBC 0 85 13.1 9 1.4 0 O.0 2 O.3 4 0.6 0 0.0 0 0.0 0 0.0 15.4 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- SEX: RALE (_) - Data UnavaiLable LABCATHE4./+3 27-JUN-2002 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-65 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID GROUP: ]-M TERMINAL CNT ABS SEX: HALE 19172 Nucleated Red Cells Ly=phocTtes Segmented Neutrophi ls Bands Monocytes Eosinophi ls Basophi Ls Abnormal Lyaphocytes Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 19173 Nucleated Red Cells W=phocytes Seglllented Neutrophi ls Bands mmocytes Eosinophlis BasophLis Abnormal L),lq_ocyt IS Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 19174 Nucleated Red Cells Lymphocytes Segmented NeutPophi Ls Bands Honocytes Eosinophi Ls Basophi Ls Abnormal Other Lymphocytes WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- (--) - Data U_vaiLabLe LABCAT HE4.43 27-JUN-2002 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal I0 GROUP: 4-M TERMINAL CNT ABS 19100 19103 19104 Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands Monocy es Eosinophi ls Basophi Ls Abnormal kymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi ls Bands e4onocyt es Eosinophi Ls Basophi Ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocyt es Segmented Neutrophi ls Bands Monocytes Eosinophi ls Basophi Ls Abnormal Lymphccyt es Other WBC 0 61 7.7 32 4.0 0 O. 0 5 O. 6 I O. I 0 0.0 1 O. 1 0 O. 0 12.6 0 78 16.4 11 2.3 0 0.0 8 1.7 0 0.0 0 0.0 3 0.6 0 O. 0 21.0 0 67 12.1 17 3.1 0 O. 0 12 2.2 2 0.4 0 0.0 2 0.4 0 0.0 18.1 418-028:PAGE K-66 SEX: MALE LABCAT HE4.43 27-dUN-2002 418-028:PAGE K-67 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID GROUP: 4-M 19118 19"133 19141 Nucleated Red Cells Lymphocytes Segmented Neutrophi ls Bands Nonocytes Eosinophi Ls Basoph i ls Abnormal Lylq:dlocy$es Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands HonocTtes Eosinophi Ls Bali Ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neut_ils Bands Nonocyt es Eosinophi Ls Basophi ls AbnormaL LymphocyIes Other WBC TERMINAL CNT ABS 0 78 9.4 12 1.4 0 O. 0 4 0.5 2 0.2 0 O. 0 4 0.5 0 O. 0 12.0 0 79 17.4 13 2.9 0 O. 0 3 0.7 4 0.9 0 O. 0 1 0.2 0 0.0 P_.0 0 73 12.7 21 3.7 0 0.0 3 0.5 2 0.3 0 O. 0 1 0.2 0 0.0 17.4 SEX: NALE LABCAT HE4._ 27-JUN-2002 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-68 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID 19142 19144 19168 GROUP: 4-M Nucleated Red Cells Lymphocytes Segmented Neutrophi ls Bands Nonocytes Eosi nophi Ls Basophi Ls Abnormal Ly._hocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands Ronocytes Eosinophi L$ k_phi ts Abnormal Other Lymphocyte= WBC Nucleated Red Cells Lymhocytes Segmented Neutrophi Ls Bands Ronocytes Eosinophi Ls Basophi Ls AbnormaL Lynlphocytes Other WBC TERMINAL CNT ABS 0 67 8.6 22 2.8 1 O. 1 8 I. 0 1 O. 1 0 O. 0 1 O. 1 0 O. 0 12.8 0 81 15.9 14 2.7 0 O. 0 3 O. 6 2 0,4 0 O. 0 0 0.0 0 O. 0 19.6 0 79 12.3 20 3. I 0 O. 0 0 O. 0 1 0.2 0 0.0 0 0.0 0 0.0 15.6 SEX: MALE LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-69 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY 140:060-069 Animal ID 19150 19156 GROUP: &-R Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands tlonocyt es Eosinophi Ls Bas,nphi Ls Abnormal Lymphocytes Other UBC NucLeated Red CelLs Lymphocytes Segmented Neutrop_i Ls Bands Monocytes Eosi nophi Ls Basophi Ls Abnormal Other gBC Lymphocytes TERflINAL cNT ABS 0 75 8.3 22 2.4 0 0.0 2 O. 2 1 0.1 0 0.0 0 0.0 0 0.0 11.1 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: HALE 19160 Nucleated Red Cells Lymphocyt es Segmented Neutrophi Ls Bands Monocles EOS_nophi Ls Basophi Ls Abnormal Lyuphocyte$ Other WBC 0 0 -- 0 -- 0 -- 0 '-- 0 -- 0 -- 0 -- 0 -- (--) - Data Unavaitable LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-70 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDYNO: 060-069 Animal ID 19162 19163 19166 GROUP:4-M Nucleated Red Cells Ly_ohocytes SegmentedNeutrophi Ls Bands Monocytes Eosi nophil s Besophils Abnormal Lymphocytes other WBC Nucleated Red Cells Lympllocytes Segmented Neutral Ls Bands Monocytes Eosi nophi ts Basophi Ls AbnormaLLymphocytes Other WBC Nucleated Red CeLls Lymphocytes SegmentedNeutrophi ls Bands Monocytes Eosinophi ls h_i Ls Abnormal Lymphocytes Other WBC TERttlNAL CNT ABS 0 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- -- SEX: MALE (_) - Data UnavaiLable LABCATHE4.&3 , 27-JUN-2002 418-028:PAGE K-71 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ID GROUP:5-M TERMINAL CNT ABS SEX: MALE 19111 Nucleated Red Cells Lymphocytes SegmentedNeutrophi Ls Bands Monocytes Eosinophi Ls Basophils Abnormal Lymphocytes Other WBC 0 83 13.0 9 1.4 0 0.0 4 O.6 1 0.2 0 0.0 3 0.5 0 O.0 15.7 19117 19124 Nucleated Red Cells LymphocyCes Secjaented Neutrophi ts Bands ltonocyces Eosinophi Ls Basophils Abnor_ L Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands ttonocy'ces EosWtophiLs Basophi Ls Abnormal Lymphocytes Other UBC 0 79 11.3 12 1.7 0 0.0 6 O.9 5 0.4 0 0.0 0 0.0 0 0.0 14.3 0 94 I0.2 3 0.3 0 O.0 2 O.2 1 O.1 0 0.0 0 0.0 0 0.0 10.8 LABCATHE4.43 27- JUN-2002 418-028:PAGE K-72 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Ani_l ID GROUP:5-M 19126 19127 19128 Nucleated Red Cells Lymphocytes SegmentedNeutrophi Ls Bands Ronocytes Eosi_i Ls Sasophi Ls Abnormal Lymphocytes Other UBC NucLeated Red CeLls Lymphocyte$ SegmentedN_t_hi ls Bands Plonocytes Eosinophi Ls Basophits Abnormal LyBphocytes Other WBC Nucleated Red Cells Ly_hocytes SegmentedNeutrophi Ls Bands l_anocytes Eosinophi ls Basophits Abnormal Lymphocytes Other WBC TERRIHAL CNT ABS 0 81 10.1 9 1.1 0 O.0 7 O.9 3 0.4 0 0.0 0 0.0 0 0.0 12.5 0 73 8.0 22 2.4 0 0.0 2 O.2 1 O.1 0 O.0 2 0.2 0 0.0 11.0 0 87 12.4 10 1.4 0 0.0 2 0.3 1 O.1 0 0.0 0 0.0 0 O.0 14.2 SEX: HALE LABCATHE4.4] 2"/'-JUN.-200_ Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal IO GROUP: 5-H TERMINAL CNT ABS 19140 Nucleated Red Cells Lywhocyt es Segmanced Neul:rophi ls hnds Monocytes Eosi_i Ls Basophi ls Abnormal tymphocytes Other WBC 0 80 12.2 14 2.1 0 O. 0 1 O. 2 2 0.3 0 O. 0 3 0.5 0 O. 0 15.2 19145 Nucleated Red Cells Lymphocytes Segmented Neutrophi ts Bands _onocyt es Eosinophi ls Basophi Ls Abnormal Lyaphocytes OCher WBC 0 83 17.2 12 2.5 0 0.0 4 O. 8 I 0.2 0 0.0 0 0.0 0 0.0 20.7 19149 Nucleated Red Cells Lylphocytes Segmented Neutrophi ts Bands MonocyCes Eosinophi ts Basophtis Abnormal Lymphocyl:es OCher WBC 0 88 16.1 12 2.2 0 0.0 0 O. 0 0 0.0 0 0.0 0 0.0 0 0.0 18.3 418-028:PAGE K-73 SEX: HALE LABCAT HE4.43 27-JUfl-2002 418-028:PAGE K-74 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 0(_)-069 Animl ID GROUP:5-H TERMINAL CNT ABS SEX: MALE 19152 Nucleated Red CeLls Lymphocytes SegmentedNeutrophi Ls Bands Monocytes Eosi nophi Ls Basophiks Abnormal Lymphocytes Other gee 0 94 13.1 3 0.4 0 0.0 3 O.4 0 0.0 0 0.0 0 0.0 0 0.0 13.9 19154 19158 Nucleated Red Cells Lymphocyl:es Segmented Neutrophi Ls Bands Monocytes Eosinophi Ls Basophi Ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lyn_hoc)rtes SegmentedNeutrophils Bands Monocytes Eosinophi Ls Basophits Abnormal Lylphocyl:es Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- (--) - Data Unavailable LABCATHE4.43 27-JUN-2002 418-028:PAGE K-75 ' Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ZD GROUP: 5-M TERMINAL CNT ABS 19168 19169 19170 Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands 14onocytes E_i_p41i Ls Basophi Ls Abnormal LymphocTtes OCher WBC Nucleated Red Cells Lymphocytes Segeented Neutnophi Ls Bands _tonocytes Eosinophi Ls sa_i ts Abnormal LymphocyCes Other WBC NucLeated Red CeLls LymphocyCes Segmented Neutrophi Ls Bands Monocytes Eosi nophi Ls Basoph JLs AbnormaL Lymphocytes Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: MALE (--) - Data Unavailable LABCAT HE4.&3 27-JUN-200_ Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-76 STUDY ID: AR6US 418-028 STUDY NO: 060-069 _imaL IO 19010 19012 19019 GROUP: 1-F Nucleated Red Cells Ly_hocyt es Segmented Neutrophi Ls Bands Monocytes Eosinophi Ls Ba_phi Ls Abnormal Lymphocytes Other UBC Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands _nocyt es Eosinophi Ls Basophi Ls Almorma I Lymphocytes Other UBC Nucleated Red Cells Lyephocytes Segmented Neutrophi Ls Bands Monocytes Eosi_i Ls Basoph i Ls Abnormal Lymphocytes Other UBC TERMINAL CNT ABS 0 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 79 12.6 17 2.7 0 0.0 3 0.5 1 0.2 0 0.0 0 0.0 0 0.0 15.9 0 66 4.4 31 2.0 0 0.0 3 0.2 0 O. 0 0 0.0 0 0.0 0 O. 0 6.6 SEX: FEHALE LABCAT HE4.43 27-JUM-2002 418-028:PAGE K-77 Study Report for Hematology WHITE DIFFERENTTAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ZD GROUP:1-F 19021 19023 19041 Nucleated Red Cells Lymphocy_ces SegmentedNeutrophi Ls Bands Monocytes Eosinophi ls Basophits Abnormal Lymphocytes Ocher MBC NucLeatedRed CeLls Lymphoc_ces SegmentedNe_r_i ls Bands Honocy_es Eosinophi Ls Basophi ls Abnormal Lymphocytes Other WBC Nucleated Red CeLLs Lymphocy1:es SegmentedNeutrophi Ls Bands Ma_ocy_es Eosinophi ts Basophi Ls Abnormal Lymphocytes Other VBC TERMZNAL CNT ABS 0 77 5.7 19 1. & 0 O.0 0 O.0 4 0.3 0 O.0 0 0.0 0 O.0 7.4 0 83 6.1 12 0.9 0 0.0 2 O.1 2 O.1 0 O.0 I O.1 0 0.0 7.3 0 63 5.0 36 2.9 0 O.0 1 O.I 0 0.0 0 O.0 0 0.0 0 0.0 8.0 SEX: FEMALE _CAT HE4.43 27-JUN-2002 418-028:PAGE K-78 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animl ID GROUP: 1-F TERMINAL CNT ABS SEX: FENALE 19042 19044 19050 Nucleated Red Cells LymphocyCes Segmented Neutr_phi Ls Bands Monocytes Eosi nophi ls Basophi ks Abnormal Lyq)hocytes Other WBC Nucleated Red CeLLs Lymlphocyt es Segmented Seutrophi Ls Bands I_onocytes Eosi nophi ls Basophi Ls N_norma L LymphocyCes other WBC Nucleated Red Cells LyBphocyCes Segmented Neut rophi Ls Bands t_nocytes Eosinophi ts Basophi ls Abnormal Ocher WBC Ly_ot_cytes 0 78 5.3 22 1.5 0 0.0 0 O. 0 0 0.0 0 0.0 0 0.0 0 O. 0 6.8 0 69 7.3 29 3.1 0 0.0 1 O. 1 1 O. 1 0 0.0 0 0.0 0 0.0 10.6 0 74 6. I 22 1.8 0 0.0 3 O. 2 I 0.1 0 O. 0 0 0.0 0 0.0 8.2 LABCAT HE4.43 27-JUN-2002 418-028:PAOE K-79 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID 19053 19065 19068 GROUP: 1-F NucLeated Red CeLLs Lymphocytes Segmented Neutrophi Ls Bands Ronocyt es Eosinophi Ls Basophi ts Abnormal lyaphocyl:es Other WBC NucLeated Red CeLLs Lylmphocyl;es Seglltented Neutrophi ls Bands Ronocytes Eosinophi Ls Bali Ls Abnormal L_c_es Other WBC Nucleated Red CeLLs Lymphocytes Segmented Neutrophi ls Bands Nonocytes Eosinophi ls Basophi Ls Abnormal Ly_hocytes Other WBC TERMINAL CNT ABS 0 71 4.7 28 1.8 0 0.0 0 0.0 0 0.0 0 0.0 I 0.1 0 O. 0 6.6 0 67 8. & 30 3.8 0 O. 0 1 O. 1 1 0.1 0 0.0 I 0.1 0 O.0 12.5 0 0 -- 0 -- 0 -- 0 h 0 -- 0 -- 0 -- 0 -- SEX: FEMALE (--) - Data Unavailable LABCAT HE4.4] 27-JUN-2002 418-028:PAGE K-80 Study Report for Hematology WEIITE DIFFER_qTIAL DATA STUDY IO: ARGUS 418-028 STUDYNO: 060-069 GROUP:1-F Animal ID TERHINAL CNT AB5 19072 Nucleated Red CeLls Lymphocyet s Segmented Neutrophi Ls Bands Monocytes Eosinophi Ls Bat_:1)Lhsi Abnormal Lyuphocyt es OCher WBC 0 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 1907+ 19075 NucLeatedRed CeLLs Lymphocytes SegmentedNeutrophi Ls Bands Hoflocytes Eosi nophi ls BasophiLs Abnoema Lymphocytes Other WBC NucLeated Red CeLLs Lymphocytes Segmented Neutrot:x_iLs Bands I_matytes Eosirm_hi Ls Basophils Ab_rma I LymphocyCes OCher WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: FEHALE (--) - Data UnavaiLabLe LABCAT HE4./+3 27-JUN-2002 418-028:PAGE K-81 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ;D 6ROUP: 2-F TERRINAL CNT ABS SEX: FEMALE 19004 19009 19016 Nucleated Red Cells LyBphocTtes SegmentedNeutrophi Ls Bands _mocy_es Eosinophi ts Basophi Ls AbnoremI Lymphocytes Other gBC Nucleated Red Cells Lyaphocytes segmented Neutrot)hi Ls Bands Ronocytes Eosinophi ts Basophits Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands Plonocytes Eosinophi Ls Basophi Ls Abnormal Lymphocytes Other UBC 0 74 5.8 25 2.0 0 0.0 1 O.1 0 O.0 0 0.0 0 0.0 0 O.0 7.8 0 87 12.7 11 1.6 0 0.0 1 O.1 1 O.1 0 O.0 0 0.0 0 0.0 14.6 0 80 13.6 18 3.1 0 0.0 0 O.0 2 0.3 0 O.0 0 0.0 0 0.0 17.0 LABCATHE4.43 27- JUN-2002 418-028:PAGE K-82 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ZD GROUP:2-F TERMINAL CNT ABS 19018 19026 190_6 NucLeated Red CeLLs Lymphocytes Segmented Neutrophi Ls Bands Honocytes Eosinophi Ls BasophiLs Abnormal Lymphocyl:es Ocher WBC NucLeated Red CeLLs Lymphocyet s SegmentedNeutrophi ts Bands Monocytes Eosinophi Ls bsophi Ls Abrmrm L Lymphocyces Ocher WBC NucLeated Red CeLLs Lymphocy'ces Segmented Neutrol_iLs Bands tt0nocytes Eosinophi Ls bsophi Ls Abnormal Ly_phoc)ret s Other WBC 0 82 10. 9 17 2.3 0 O.0 0 O.0 I 0.1 0 0.0 0 0.0 0 0.0 13.3 0 85 7.7 15 q.4 0 0.0 0 O.0 0 0.0 0 0.0 0 O.0 0 0.0 9.1 0 84 15.1 12 2.2 0 0.0 2 O.4 2 O.4 0 O.0 0 0.0 0 O.0 18.0 SEX: FENALE LABCATHE4.43 27- JUN-2(X)2 418-028:PAGE K-83 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ID GROUP:2-F 19037 19043 19047 NucLeated Red CeLLs Lymphocytes Segmented Neutrophi Ls Bands Renocytes Eosinophi Ls BasophiLs AbrmPmL tymphocytes Ocher WBC NucLeated Red CeLLs Lymphocytes SegmentedNeurtrophits Bands Ronocy'tes Eosinophi ls BasophiLs AbnOllaL tSnmphocytes Other WBC NucLeated Red CeLLs Lylphocytes Segmented Neutrophi Ls Bands Monocytes Eosinophi Ls BesophiLs Abnormal Lymphocytes Other WBC TERfllNAL C.T ABS 0 42 11.0 52 13.7 0 0.0 6 1.6 0 0.0 0 O.0 0 0.0 0 0.0 26.3 0 71 9.6 26 3.5 0 O.0 2 0.3 1 0.1 0 0.0 0 0.0 0 0.0 13.5 0 56 6.2 41 A.6 0 O.0 2 0.2 I 0.1 0 O.0 0 0.0 0 O.0 11.1 SEX: FEHALE LABCATHE4.43 27- JUN-2002 418-028:PAGE K-84 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS /+18-028 STUDY NO: 060-069 AnimaL ZD 190_ 19052 19055 GROUP: 2-F Nucleated Red CelLs Lymphocytes Segmented Neutrophi Ls Bands Honocyzes Eosinophi ls 6asophi Ls Abnormal Lymphocytes Other gBC Nucleated Red Cells Lylq)hocytes Segmented NeUtrophi Ls Bands Monocytes Eosinophi ts Baeol_i Ls Abnormal OCher tymphocytes WBC Nucleated Red Cells Lymphocyt es Secjlented Bands Neutrophi Ls Ronocytes Eosi nophi Ls Basophi ts Abnormal Lymphocytes Ocher WBC TERHINAL CNT ABS 0 76 5.4 23 1.6 0 0.0 0 O.0 1 0.1 0 O. 0 0 0.0 0 0.0 7.1 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: FEMALE (_) - Data UnavaitabLe LABCAT HE4.43 27-JUN'-2002 418-028:PAGE K-85 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYlO: ARGUS418-028 STUDYNO: 060-069 AnilmL ID GROUP:2-F TERHINAL CNT ABS 190(51 19067 Nucleated Red Cells Lymphocyres Segmented NeutrophiLs Bands HormcyCes Eosinophi Ls Basophi Ls Abnormal ky_ho_Ttes OCher WBC NucLeated Red CeLLs Lymphocytes Segmented Neutrophi Ls Bands Monocytes Sosinophi Ls BasophiLs Abnormal Lymphoc)rtes Other WBC O 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- N 19071 Nucleated Red Cells Lymphocytes Segmented NeutrophiLs Bands Ronocyces Eosinophi ls Basophits Abnormal L),aphocytes Other tJBC O 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: FEMALE (--) - Data UnavaitabLe I._kT HE4.43 27- JUN-20_ Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 GROUP:_-F Animal ID TERMINAL CNT ABS 19003 NucLeated Red CeLLs Lymphocytes St_3mentedNeutrophi Ls Bands Monocytes Eosirw)phiLs EmsophiLs Abnormal byeq_ho_tes Other WBC 0 59 6.5 36 4.0 0 O.0 3 0.3 1 0.1 0 0.0 1 0.1 0 0.0 11.1 19007 NucLeated Red CeLLs Lymphocy_es Se_mte(I Bancls Neutrc_YaLi s Honocytes Eosinophi ts Basophi Ls AbnormaLLymphocytes Other WBC 0 72 5.7 25 Z.O 0 0.0 2 0.2 0 0.0 0 O.0 1 0.1 0 0.0 7.9 19008 NucLeated Red CeLLs Lymptmc)_es Segmented NeutrophiLs Bands eW=nocytes Eosi nophi Ls Basophi Ls AbnormaL L)n_hocytes Other WBC 0 72 5.0 27 1.9 0 0.0 0 O.0 0 O.0 0 0.0 1 0.1 0 0.0 7.0 418-028:PAGE.K-86 SEX: FEMALE "LABCATHE4.4] 27-JUN-2002 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-87 STUDY ZD: ARGUS 418-028 STUDY NO: 060-069 AnimL I0 19013 19014 19015 GROUP: 3-F Nucleated Red Cells Ly_phocyt es Segaented Neut rophi ts Bands Honocytes Eosinophi Ls Bas_phi ls N:morua L kyaphocyl;es Other WBC Nucleated Red Cells L_phocytes Segmented Neutrophi ls Bands Honocytes Eos_ nophi ts Basophi Ls Abrmrlm I Lymphocytes Other WBC Nucleated Red Cells Lymld_ocytes segmented Neutrophi Ls Bands Monocy_es Eosinophi Ls Basophi L= Abnormal Ly-mphocyl:es Ocher WBC TERMINAL CNT ABS 0 7& 12.3 23 3.8 0 O. 0 2 O. 3 0 0.0 0 0.0 1 0.2 0 0.0 16.6 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 77 5.7 22 1.6 0 0.0 1 O. 1 0 0.0 0 0.0 0 0.0 0 0.0 7.4 SEX: FEMALE (--) - Data Unavailable LABCAT HE4._ 27-JUN-2002 418-028:PAGE K-88 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ID GROUP:]-F TERMINAL CNT ASS SEX: FEMALE 19017 Nucleated Red Cells LymphocyCes SegmentedNeutrophi Ls Bands Monocy_es Eosinophi Ls Basophi Ls Abnormal Lymphocytes O_d_er WBC 0 76 9.8 20 2.6 0 0.0 1 O.1 5 0.4 0 0.0 0 0.0 0 0.0 12.9 19024 Nucleated Red Cells Lymphocyl:es Segeented Neutrc_ohiLs Bands Nonocytes Eosinophi Ls Basophi ts Abnormal Lymphoc)rets other WBC 0 70 7.4 25 2.6 0 0.0 2 O.2 2 0.2 0 0.0 1 0.1 0 0.0 10.5 19029 Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands l_nocytes Eosi_i Ls Basophi ls Abnorw L Ly.q_hocyCes Other WBC 0 82 10.3 17 2.1 0 0.0 0 O.0 I 0.1 0 O.0 0 0.0 0 0.0 12.5 LABCATHE6.43 27-JUN-2002 418-028:PAGE K-89 ' Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY NO: 060-069 AnimaL .T.D 6ROUP: 3-F TERHINAL CHT ABS SEX: FEMALE 19034 Nucleated Red Cells Lymphoc)rtes Segmented Neutrotot_i ls Bands Ronocytes Eosi _i Ls Basophi ls Abnormal Lyuphocytes Other WBC 0 90 10. 9 8 1.0 0 O. 0 1 O. 1 I O. 1 0 O. 0 0 0.0 0 O, 0 12.1 19038 1_)56 Nucleated Red CeLLs Lymphoc)rtes Segmented Neutrophi ls Bands Honoc_ces Eosinophi Ls bsophi ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented tteut rophi Ls Bands _nocytes Eosi _hi ls Basophi ls Abnormal LymphocyCes Other WBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 80 I0.2 17 2.2 0 O. 0 3 O. 4 0 O. 0 0 0.0 0 0.0 0 0.0 12.7 (--) - Data UnavailabLe LABCAT HE4.43 27-JUN-2_ Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-90 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal ID 19057 19060 19064 GROUP: 3-F Nucleated Red Cells Lymqohocytes Segmented Neutr_hi ls Bands _tonocytes Eosinophi ls Basophi ls Abnormal Other WBC Lymmphocy_es Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands Monocy_es Eosi nophi ls bsophi ts Abnormal Lymphoc)rt es Other WBC Nucleated Red Cells Lyaphocyt es Segmented Neutrophi Ls Bands Honocy_es Eosinophi ls Baso_i Ls AbnormaL Ly:phocytes Other WBC TERMINAL C_ _S 0 0 -- 0 -- 0 -- 0 -- 0 -- 0" -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: FEMALE (--) - Data UnavaiLabLe LABCAT HE4. _3 27- JUN-2002 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-91 STUDY ID: ARGUS 41H28 STUDY NO: 060-069 Animal ID GROUP: &-F TERMINAL CNT ABS SEX: FERALE 19002 Nucleated Red Cells Ly_0hocyces Segmented Neutrophi ls Bands Honocytes Eosi nophi Ls Bas_hi ls A_rmal Other WBC Ly_cytes 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 19005 Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands Nonocyces Eosinophi ls Basophi Ls Abnormal Lymphocyt es Other WBC 0 81 7.7 17 1.6 0 O. 0 1 O. 1 I 0.1 0 0.0 0 0.0 0 O. 0 9.5 19035 Nucleated Red Cells Lymphocytes Segmented Neutrophi ls Bands HonocyT es Eosinophi ls Basophi ls /d_rmal Lymphocytes Other gSC 0 7/, 7.1 24 2.3 0 0.0 1 0.1 0 0.0 0 0.0 1 O. 1 0 0.0 9.6 LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-92 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUOYNO: 060-069 Animal ZD GROUP:4-F TERtIINAL CNT ABS SEX: FEHALE 19039 19040 19045 Nucleated Red Cells Lymphocytes Segmented Heutrophi ls Bands e4onocyet s Eosinophi Ls Bali ts Al_orma l Lymphocytes Other WBC NucLested Red Cells Lyaphocyl:es Segmented Neutrophi ls Bands Monocytes Eosinophi ls Em_phi ls Abnormal Lymphocytes Other UBC Nucleated Red CelLs Lym_hocytes Segmented Neurtrophi ls Bands Ronocytes Eosinophi Ls Basophi Ls Abnormal Ly_ohocytes Other WBC O 75 9.6 22 2.8 0 0.0 2 0.3 1 O.1 0 0.0 0 O.0 0 0.0 12.8 0 77 7.7 22 2.2 0 0.0 1 O.1 0 0.0 0 0.0 0 0.0 0 0.0 lO.0 0 56 5.4 39 3.8 3 0.3 2 0.2 0 0.0 0 0.0 0 0.0 0 O.0 9.7 (--) - Data UrmvailabLe L/_CAT HE4._ 27- JUN-2(X)I2 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-93 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal 1D GROUP: 4-F TERMINAL CNT ABS SEX: FEMALE 19046 19051 19054 Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands )k_nocytes Eosi nophi Ls Basophi Ls Abnormal Lymphocytes Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi Ls Bands e_ocytes Eosinophi ls Basophi ls Abnormal Other Lymphocyt es WBC Nucleated Red Cells LymphocTtes Segamnted Neutrophi ls Bands Honocytes Eosinophi Ls Basophi ls Abnormal Lymphocytes Other gBC 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 82 12.2 13 1.9 0 O. 0 ] O. 4 0 0.0 0 0.0 2 0.3 0 O. 0 14.9 (--) - Data Unavailable LABCAT HE4.4] 27- JIJN-2002 418-028:PAGE K-94 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYI0: ARGUS418-028 STUDYNO: 060-069 Animal ID GROUP:&-F TERMINAL CNT ABS SEX: FEMALE 19058 19062 19063 Nucleated Red Cells Lymphocytes SegmentedNeutrophi Ls Bands Monocytes Eosinophi Ls BasophiLs AbnormaL Lymphocytes Other WBC Nucleated Red Cells ty_ocyt es SegmentedNeutrophi Ls Bands MonocyCes Eosinophi Ls BasophiLs AimorBaL LyBphocytes Other WBC NucLeated Red CeLLs Lymphocyet s Segmnzed Neutrophi Ls Bands Honocytes Eosinophi Ls BasophiLs Abnormal Lymphocytes Other UBC 0 73 11.5 25 3.9 0 O.0 1 0.2 1 0.2 0 0.0 0 0.0 0 0.0 15.7 0 77 4.8 22 1.4 0 O.0 1 O.1 0 0.0 0 0.0 0 0.0 0 0.0 6.2 0 76 9.8 20 2.6 0 0.0 2 O.3 2 0.3 0 O.0 0 0.0 0 0.0 12.9 LABCATHE&.43 27- JUN-2OO2 418-028:PAGE K-95 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS418-028 STUDYNO: 060-069 Animal |D GROUP:4-F 19066 19069 1907"5 NucLeated Red CeLLs Ly_ahocytes SegmentedNeutrophits Bands I_ocyt es Eosinophi Ls BasophiLs AbnormmLky_:hoc_r:es Other WBC Nucleated Red Cells Lymphocytes Segmented Neutrophi ts Bands l_ocytes Sosinophi Ls Sasophi ts Abnormal Lyalphocyl:es Other YBC NucLeated Red CeLLs LyWnocytes Segmented Neu_rophi ls Bands Nonocytes EosinophiLs Basophi Ls Abnormal Lymphocy_ces Other WEC TERMINAL CNT ABS 0 84 6.0 14 1.0 0 O.0 I O.1 0 0.0 0 0.0 1 0.1 0 0.0 7.1 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- SEX: FEMALE (--) - Data UnavaiLabLe LABCATHE4.4] 27- JUN-2OQ2 Study Report for Hematology WHITE DIFFERENTIAL DATA 418-028:PAGE K-96 STUDY ID: ARGUS 418-028 STUOY NO: 060-069 Ani_l ID GROUP: 5-F TERMINAL CNT ABS SEX: FERALE 19001 19006 Nucleated Red Cells Lylphoc)rt es Segmented Neutrophi ls Bands _onocy_ es Eosi nophi Ls _Ql_hi Ls _r_l L)qlphocytes Other WBC Nucleated Red Cells Lymphocytes Seglmflted Neutrophi ls Bands Honocytes Eosinophi ls _i Ls Abnormal tyllphocytes Other UBC 0 67 10.1 28 4.2 0 0.0 3 0.5 2 O. 3 0 O. 0 0 0.0 0 O.0 15.1 0 61 4.6 36 2.7 0 0.0 3 0.2 0 0.0 0 0.0 0 0.0 0 0.0 7.6 19011 Nucleated Red Cells LyaphocyCes Segmented Neutrophi Ls Bands Honocytes Eosinophi ls Basophi ts Abnormal Other Lymphocytes WBC 0 69 4.4 28 1.8 0 0.0 2 O. 1 0 0.0 0 0.0 1 O. 1 0 0.0 6.4 LABCAT HE4.1_ 27-JUN-2(X}2 418-028"PAGEK-97 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ZD: ARGUS 418-028 STUDY NO: 060-069 Animal Ii) GROUP: 5-F 19020 19022 19025 Nucleated Red Cells Lymphocytes S_m_ted Bands N_utr_i ls I_nocytes Eosinophi ls Ba_hi ls Abnormal Lymphocyt es Other WBC Nucleated Red CelLs Lymphocy_es segmented Neut_i Ls Bands NonocyCes Eosinophi ls ' Basophi Ls _rmaL Other Lyephocytes WBC NucLeated Red CeLLs Lymphocy_ es Segmanted Neutrophi Ls Bands Honoc)rtes Eosi nophi ts Bas_hi Ls Abnormal Lymphocyt es Other WBC TERMINAL CNT ABS 2 52 2.2 46 1.9 0 0.0 2 0.1 0 0.0 0 0.0 0 0.0 0 0.0 4.2 0 58 3.1 37 2.0 0 O. 0 3 0.2 2 0.1 0 0.0 0 0.0 0 O. 0 5.4 0 71 5.2 27 2.0 0 0.0 1 O. 1 0 0,0 0 0.0 1 0.1 0 0.0 7,3 SEX: FEMALE LABCAT HE4.43 27-JUN-2002 418-028:PAGE K-98 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDYID: ARGUS418-028 STUDYNO: 060-069 AnimoL II) GROUP:5-F TERI4IHAL CNT ABS SEX: FEHALE 19027 19028 Nucleated Red CeLLs Lymphocyl:es SegmentedNeutrophi ls Bands Monocytes Eosi nophiLs Basophi Ls A_rmma I t)qaphocyZes Other WBC NucLeated Red CeLLs Lylmphocytes Segmented Neutrophi ts Bands Nonocytes Eosinophi ls Basophi ts A_rmal Lympllocytes Other WBC 0 8] 10.2 16 2.0 0 O.0 1 O. 1 0 0.0 0 O.0 0 0.0 0 0.0 12.3 0 70 7.1 27 2.8 0 O.0 3 0.3 0 0.0 0 0.0 0 0.0 0 0.0 10.2 19030 Nucleated Red CeLLs Lymphocytes Segmnzed Neutr_i ts Bands Ronocytes Eosinophi Ls Basophi ts Abnormal Lymphocytes Other WBC 0 64 5.2 34 2.8 0 0.0 1 0.1 0 0.0 0 0.0 1 0.1 0 0.0 8.1 LABCATHE4.43 27-JUN-L:_)02 418-028:PAGE K--99 , Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS 418-028 STUDY 110:060-069 AnimL ID GROUP: 5-F TERMINAL CNT ABS SEX: FEMALE 19051 19032 NucLeated Red CeLLs LyBphocyt es Segmented Neutrophi ls Bands Monocy't es Eosi nophi ls Basophi ts Abnormal LyBphocyt es Other WBC NucLeated Red CeLLs Lymphocyt es Segmented Neutrophi Ls Bands Monocytes Eosi _i ls _i ts AbnormaL Lymphocytes Other gBC 0 67 8.2 30 3.7 0 O. 0 0 O. 0 5 0.4 0 O. 0 0 0.0 0 O. 0 12.5 0 0 w 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- -- 19035 Nucleated Red Cells Ly_ohocytes Segmented Neutrophi ls Bands Monocytes Eosi nophi ts Basophi ls Abnormal Lymphocytes Other WBC 0 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- (--) - Data Unavailable LABCAT HE4.45 27-JUN-2002 418-028:PAGE K-100 Study Report for Hematology WHITE DIFFERENTIAL DATA STUDY ID: ARGUS418-028 STUDYNO: 060-069 Animal ID 19049 GROUP:5-F Nucleated Red Cells Lymphocytes Segmented heutrophils Bands Monocytes Eosinop_i ls hsophi ls Abnormal Lymphocy_es Other TERPtINAL CNT ABS 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- SEX: FEtlALE 19059 19070 NucLeated Red Cells Lye_ocytes S_a_nted Neut_i ls Honocytes Eosinophi Ls Basophils AbnoreaI Lymphocytes Other MBC Nucleated Red Cells Lymphocytes SegmentedNeutrophiLs Bands l_nocytes Eosinophi Ls BasophiLs Abnormal Lyaphocy_es Other WBC 0 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- 0 -- (--) - Data Unevai lable LABCATHE4.43 27-JUN-2(X]2 418-028:PAGE K-101 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT PERIOD: TERMINAL BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: RAtE Animal ID GROUP: 1-R 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 19143 19151 19157 19161 19167 MEAN SO N TP g/dL ALB g/dC 5.9 3.9 6.5 4.4 5.8 4.0 6.0 4.1 6.2 4.3 6.5 4.3 6.5 4.2 6.1 4.3 6.3 4.3 7.0 4.7 ......... ......... .......... ........ ......... 6.3 0.36 10 4.3 0.22 10 GLU mg/dL 164 165 119 134 140 169 128 141 140 153 145 16.8 10 CHOL ,,g/dL 46 62 61 44 62. 56 59 51 56 72 57 8.3 10 T-BZL mg/dL 0.1 0.2 0.2 0.1 0.1 0.3 0.1 0.1 0.2 0.2 0.2 0.07 10 BUN mg/dL 16 19 16 14 15 15 18 15 15 15 CREAT mg/dL 0.3 0.4 0.3 0.3 0.3 0.4 0.3 0.3 0.3 0.3 CK U/L 263 164 330 105 177 260 385 250 722 221 16 0.3 288 1.5 0.04 172.6 10 10 10 (--) - Data UnavaiLable LABCAT C4.43 ZT-JUN-2002 418-028:PAGE K- 102 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL BY GROUP STUDY IO: ARGUS 41H28 STUDY NO: 060-069 Animal ID GROUP: 2-M 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 19147 19153 19164 19165 19171 TP g/dL ALB g/dL 6.1 4.1 6.2 4.0 5.8 3.9 6.1 4.1 5.6 3.9 6.5 4.3 5.7 3.9 6.2 4.3 6.3 4.4 6.1 4.2 .......... ........ ........ ......... ......... MEAN SD N 6.1 0.28 10 4.1 0.19 10 GLU IIIg/dL 172 166 152 104 157 115 149 123 108 165 141 26.0 10 CHOL mg/dL 40 44 38 :33 30 28 _3 52 60 55 T-BIL mg/dL 0.2 0.1 0.1 0.1 0.1 0.2 0.1 0.1 0.2 0.2 41 11.1 10 O. I 0.05 10 BUN mg/dL 16 17 15 16 16 17 17 17 15 16 CREAT mg/dL 0.3 0.3 0.3 0.3 0.3 0.4 0.3 0.3 0.4 O.& SEX: MALE CK U/L .169 151 235 307 133 350 292 134 478 213 16 0.3 246 0.8 0.05 111.5 10 10 10 (--) - Data UrmvailabLe LABCAT CC4.43 27-JUN-2002 418-028:PAGE K- 103 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal tO GROUP: 3-M 19101 19105 19107 19112 19114 19121 19123 19130 10137 19146 19155 19159 19172 19173 19174 MEAN SD N TP g/dL ALB g/dL 6.2 H 6. I 5.8 6.3 6.3 6.6 6.2 5.7 6.7 6.6 .......... ......... ........ .......... ......... 4.0 H 4.3 4.1 4.3 4.3 4.4 4.4 4.1 4.4 4,4 6.3 0.33 10 4.3 0.15 10 GLU lmg/dL 304 H 254 198 137 142 121 156 149 158 204 182 58.2 10 CHOL mg/dL 27 H 52 50 46 65 45 46 34 59 35 46 11.6 10 T-BIL mg/dL 0.3 H 0. I 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.06 10 BUN mmg/dL 18 H 16 15 12 16 18 17 17 17 16 16 1.8 10 CREAT mg/dL 0.4 H 0.3 0.3 0.3 0.3 0.4 0.2 0.3 0.3 0.3 SEX: MALE CK U/L 129 H 118 132 286 250 289 329 480 200 175 0.3 0.06 10 239 112.9 10 (--) - Data Unavaitable LABCAT CC4.43 H - HemoLyzed 27-JUN-2002 418-028:PAGE K-104 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT PERIOD." TERMINAL BY GROUP STUOY 10: ARGUS 418-028 STUDY NO: 060-069 SEX: MALE Animal ID GROUP: 4-M 19100 19103 19104 19118 19133 19141 19142 19144 19148 19150 19156 19160 19162 19163 19166 MEAN SP N TP g/dL ALB g/dL 6.0 4.0 5.7 4.0 5.9 4.1 6.0 &.2 6.3 4.4 6.2 4.3 6.6 4.4 6.2 4.1 6.5 4.4 6.4 4.4 ......... ......... ......... ........ ......... 6.2 0.28 10 4.2 0.17 10 GLU mj/dL 175 202 167 131 179 147 132 130 175 181 162 25.2 10 CHOL =g/dL 42 28 42 39 49 70 56 29 42 30 T_IL mg/dL 0.1 0.1 0.1 0.1 0.1 0.2 0.2 0.1 O.1 0.2 BUN -g/dL 16 20 16 20 17 16 17 18 16 18 43 0.1 17 13.1 0.05 1.6 10 10 10 CREAT ,_/dL 0.3 0.3 0.3 0.4 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.03 10 CK U/L 111 145 196 187 146 182 270 242 267 218 196 53.5 10 (--) - ORtm UrmvaiLabLe LABr.AT C4.43 27-JUN,'-2002 418-028:PAGE K-I.05 Study Report for Clinical INDIVIDUAL ANIMAL REPORT PERIOD: TERMINAL Chemistry BY GROUP STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Animal %O GROUP: 5-M 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 19154 19158 19168 19169 191"0 TP g/dL AN g/dL 6.6 H 6.1 H 6.6 6.4 6.6 6.5 6.0 6.4 6.3 5.8 ........ ......... ......... ......... ........ 4.6 H 4.3 H 4. B 4.6 4.6 4.8 4.3 4.5 4.5 4.2 MEAN SD N 6.3 0.28 10 4.5 0.20 10 GLU i_j/dL 173 H 177 H 226 165 156 181 110 134 195 132 165 33.7 10 CHOL I_J/dL 38 H 37 H 25 29 47 35 24 27 38 31 33 7.2 10 T-BIL mg/dL 0.2 H 0.3 H O. 1 O. 1 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.06 10 BUN mg/dL 26 H 2.3 H 20 20 22 21 18 19 19 19 21 2.4 10 CREAT l_j/dL 0.4 H 0.3 H 0.4 O. 3 0.3 0.4 0.3 0.3 0.3 0.3 SEX: MALE CK U/L 211 H 269 H 94 156 379 472 304 239 175 128 0.3 0.05 10 243 117.4 10 (--) - Data UnavaitabLe LABCAT CC4.43 H - Hemmotyzed 27-JUN-2002 418-028:PAGE K-106 Study Report for Clinical Chemistry INDMDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDYID: ARGUS418-028 STUDYNO: 060-069 AnimaL ID GROUP:1-F 19010 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 19074 10075 TP g/dL ALB g/dC ........... 5.4 3.7 6.3 4.3 7,1 4.9 6.1 4.3 5.9 4.3 6.3 4.3 6.6 4.4 5.8 3.9 5.5 3.7 5.7 3.5 ........ ........ ........ .......... MEAfl SD N 6.1 0.52 10 4.1 0.42 10 GLU I_/dL 149 139 157 156 165 155 144 184 106 163 152 19.8 10 CHOt ag/dC 63 92 136 85 62 77 93 78 43 40 N 27.8 10 T-BIL KJ/dL 0.2 0.1 0.1 0.2 0.2 0.1 0.3 0.2 0.1 0.1 O.Z 0.07 10 BUN sg/dL SEX: FEMALE CREAT _J/dL CK U/L 32 0.4 166 29 0.3 212 33 0.4 210 26 0.4 222 28 0.3 201 25 0.4 178 27 0.4 332 34 0.3 198 28 0.3 123 19 0.3 175 28 0.4 202 4.4 0.05 54.1 10 10 10 (--) - Data Unavailable LABCATCC4.43 27-JUN-2002 418-028:PAGE K-107 Study Report for Clinical INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL Chemistry BY GROUP STUDY IO: ARGUS41H28 STUOYN0:060-069 Animal IP GROUP: 2-F 19004 19009 19016 19018 1907.6 19036 19037 19043 19047 19048 10052 19055 19061 19067 19071 MEAN SO N TP g/dL AL_ g/dL 5.9 4.3 5.7 3.9 5.5 4.2 5.6 4.2 5.5 3.9 5.9 4.2 5.O 3.5 5.7 3.8 4.3 3.3 5.9 4.2 ......... ........ ........ ........ ........ 5.5 0.50 10 4.0 0.34 10 GLU mg/dL 178 166 173 152 170 203 153 149 135 142 162 20.0 10 CHOL q/dL 56 62 83 77 85 B4 54 92 57 73 T-elL mgldL 0.2 0.2 0.2 0.2 0.1 0.2 0-2 0.2 0.2 0.2 72 14.0 10 0.2 0.03 10 BUN mcjldL 20 26 26 27 32 22 ?..2 21 16 29 24 4.7 10 SEX: FERALE CREAT mg/dL 0.3 0.4 0.3 0.4 0.3 0.4 0.3 0.4 0.3 0.4 CK UIL 144 311 ?..37 151 208 22.8 369 269 392 186 0.4 0.05 10 250 85.6 10 (--) - Data UnavaiLabLe LABCATCC4.43. 27-JUfl--2002 418-028:PAGE K- 108 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 SEX: FEMALE Animal ID GROUP: 3-F 19003 19007 19008 19013 19014 19015 19017 19024 19029 19054 19038 19056 19057 19060 19064 MEAN SP N TP O/dL AIra g/dL 5.6 3.9 5.3 3.8 5.7 4.0 5.8 4.1 ........ 5.6 3.7 5.8 4.1 6.7 4.5 5.8 4.0 6.& 4.3 ........ 6.1 4.2 ......... ........ ....... 5.9 0.41 10 4.1 0.24 10 GLU mg/dL 179 121 171 146 155 164 140 142 151 179 155 18.7 10 CHOL mg/dL 54 45 56 59 61 87 96 79 105 63 "}1 20.0 10 T-BIL mcJ/dL 0.1 0.1 0.1 0.2 0.2 0.2 0.2 0.2 0.1 0.1 0.2 0.05 10 BUN mg/dL 56 41 20 24 28 22 33 26 23 28 28 6.7 10 CREAT mcJ/dL 0.3 0.3 0.3 0.4 0.3 0.4 0.4 0.3 0.4 0.3 0.3 0.05 10 CK U/L 199 159 206 160 212 190 215 212 169 197 -- 192 21.7 10 (--) - Data UnavailabLe LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-109 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL STUDY/D: ARGUS&18'-028 STUDYNO: 060-069 SEX: FEHALE Animal ID GROUP:4-F 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058 19062 19063 19066 19069 19073 TP g/dL ALB g/dL ......... 6.1 4.3 5.7 4.0 5.5 4.0 5.8 3.9 5.6 3.7 ........ ........ 6.3 4.5 5.6 3.9 6.0 4.4 5.2 3.6 5.9 4.1 ........ ........ GLU mg/dL HOL IKj/dL T-BTL Imj/dL 167 70 0.2 161 47 0.1 163 71 0.2 127 63 0.1 160 91 0.2 129 41 0.1 150 64 0.1 127 49 0.1 145 62 0.1 132 63 0.1 BUN IIg/dL CREAT IKj/dL 30 0.4 27 0.4 26 O.& 20 0.3 25 0.5 25 0.4 20 0.3 34 0.3 23 0.4 22 0.2 CK U/L 259 193 160 253 398 124 267 176 172 147 MEAN SO N 5.8 0.32 10 4.0 0.29 10 146 16.2 10 62 14.2 10 0.1 0.05 10 25 0.4 4.4 0.08 10 10 213 80.4 10 (--) - Data UnavaiLabLe LABCATCC4.43 27- JUlt-2002 418-028:PAGE K-110 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL STUDYID: ARGUS418-028 STUDYNO: 060-069 Artist ID TP _ g/dL g/dL GROUP:5-F 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 19049 19059 19070 5.5 4.0 5.7 4.0 5.8 4.3 5.6 3.5 5.8 4.1 5.8 3.7 5.5 3.8 5.9 4.0 6.3 4.5 6.3 4.4 .......... ........ ........ ........ ........ MEAN SI) N 5.8 0.29 10 4.0 0.31 10 Gt.U lg/dL C_L mg/dL 165 55 119 58 154 48 53 33 132 60 133 49 152 71 159 70 180 58 157 108 T.-SU. mg/dL 6u. ag/dL 0.1 23 0.1 19 0.1 29 O.2 73 0.1 25 0.1 28 O.1 27 0.2 ?.3 0.1 26 0.2 25 SEX: FEMALE CREAT mg/dL CK U/L 0.3 341 0.3 373 0.4 149 O.2 438 0.3 186 0.4 143 O.3 252 0.3 261 0.3 168 0.3 210 140 35.5 10 61 19.8 10 0.1 0.05 10 30 15.4 10 0.3 0.06 10 252 101.6 10 (--) - Data UnavaiLabLe LABCATCr_./_3 27-JUN-2002 418-028:PAGE K'-I11 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD = TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 AnimaL ID GROUP: 1-M 19109 19115 19116 19119 19122 19125 19131 19132 19134 19135 19143 19151 19157 19161 19167 ALT AST U/L U/L 36 78 38 77 40 93 33 83 51 86 49 113 39 116 42 80 49 146 43 85 ........... ........ ......... ........ ........ MEAN SD N 42 96 6.0 22.4 10 10 ALP U/L 91 77 102 112 103 115 107 98 126 119 105 14.3 10 CA IJ/dL 10.6 11.8 10.3 10.8 11.3 10.6 11.1 11.1 10.4 11.4 PHOS I_/dL 12.0 11.6 9.7 7.9 9.& 8.? 7.9 9.B 8.5 9.2 TRIG I_/dL 2:3 48 59 43 28 65 93 45 36 76 NA mmo',./L 143 146 145 147 146 1/,5 146 146 147 148 SEX: MALE K mmot/k 10.3 10.5 6.6 4.6 5.9 5.5 4.9 6.2 5.7 5.5 10.9 0.48 10 9.5 1.40 10 52 ;)I.8 10 146 6.6 1.4 2.10 10 10 (N) _ Data Unavai LabLe LABCAT CC4.43 27-JUN-200_ Study Report for Clinical INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL Chemistry BY GROUP 418-028:PAGE K-112 STUDYID: ARGUS4t8-028 STUDYNO: 060-069 SEX: MALE Animal IO GROUP:2-M 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 19147 19153 19164 19165 19171 ALT AST U/L U/L 37 84 38 77 37 81 57 116 40 91 46 111 48 104 3/, 80 261 382 33 87 ........ ........ ......... ......... ........ ALP U/L 101 70 99 141 130 91 99 88 199 95 CA Ig/dL 10.5 11.6 10.3 10.3 11.2 10.7 10.6 10.7 10.3 11.1 PHOS _J/dL 10.3 12.0 9.1 8.9 8.9 9.3 9.3 8.8 7.8 9.3 TRIG mg/dL 26 60 62 44 43 29 27 78 57 43 NA =,,oL/L 145 144 145 146 147 146 145 144 147 148 K "=_/L 6.8 8.0 6.0 5.4 5.2 5.5 6.2 6.9 5.5 5.5 MEAN SO N 63 69.9 10 121 92.6 10 111 36.9 10 10.7 0.44 10 9.4 1.11 10 47 17.2 10 146 6.1 1.3 0.89 10 10 (--) - Data UnavailabLe LABCATCC4.43 27- JUN-210_ 418-028:PAGE K-113 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL STUDYID: ARGUS418-028 STUDYNO: 060-069 SEX: HALE Animal ID GROUP:3-M 19101 19105 19107 19112 19114 19121 19123 19130 1913'7 19146 19155 19159 19172 19173 19174 ALT AST U/L U/L 60 H 35 35 45 141 42 101 55 44 42 ......... ........ ........ ........ ........ 12/, H 70 82 99 197 92 171 140 102 94 ALP U/L 101 H 107 97 92 108 88 80 124 108 96 CA ,m/dE 11.5 H 11.5 11.5 10.2 10.8 10.7 11.2 10.7 11.3 11.3 PHOS _J/dL 10.4. H 9.5 15.6 8.8 9.9 9.0 9.4 9.4 8.3 8.8 TRIG mg/dL NA ==ot/L K mmoL/L 20 H 56 34 51 35 56 21 34 ?2. 35 1&3 H 147 148 147 145 147 146 148 146 148 8.9 H 5.6 8.4 6.5 7.0 5.5 6.1 5.4 5.7 5.4 REAN SD N 60 34.4 10 117 40.9 10 100 12.4 10 11.1 0.44 10 9.9 2.09 10 36 13.8 10 147 6.5 1.6 1.27 10 10 (--) - Data UnavaiLable LABCATCC4.43 H - Hemolyzed 27-JUN-2002 418-028:PAGE K-114 ' Study Report for Clinical Chemistry INDIVIDUAL _ PERIOD: REPORT BY GROUP TERMINAL STUDY ID: ARSUS 418-028 STUDY NO: 060-069 SEX: MALE Animal ID GROUP: 4-M 19100 19103 19104 19118 19133 19141 19142 1914/, 19148 19150 19156 19160 19152 19163 19166 MEAN SD N ALT AST U/L U/L 51 91 37 83 50 107 439 887 54 106 46 9'I 45 108 48 119 138 277 46 107 ........ ........ ........ ........ ......... 95 124.1 10 198 248.7 10 ALP U/L 122 76 98 154 152 93 98 124 112 123 115 25.1 10 CA mg/dL 10.9 11.0 11.5 10.8 11.7 10.T 10.7 11.2 11.2 11.3 11.1 0.34 10 PHOS mg/dL 8.3 13.3 8.6 9.8 9.3 10.7 8.8 9.8 9.2 9.3 TRIG mg/dL 14 19 35 23 32 113 35 30 28 32 NA maol/t, 146 147 148 148 147 145 148 147 147 146 K emmot/L 5.5 6.6 5.6 6.9 5.3 6.4 5.3 5.5 6.1 6.5 9.7 1.43 10 36 27.9 10 147 6.0 1.0 0.60 10 10 (--) - Data Unavai LabLe LABCAT CC4.43 27-JUN-20(]2 418-028:PAGE K-115 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDY ZD: AR6US 418-028 STUDY NO: 060-069 SEX: MALE Animal IO GROUP: 5-M 19111 19117 19124 19126 19127 19128 191&0 19145 19149 19152 19154 19158 19168 19169 19170 MEAN SO N ArT AST UIL U/L 38 H 49 H 33 52 49 50 40 54 40 42 ......... ........ ........ ......... ........ 86 H 114 H 76 88 104 112 95 125 87 87 45 97 7.0 15.6 10 10 ALP U/L 115 H 153 H 204 159 182 83 169 137 140 99 144 37.5 10 CA "g/UL 11.7 H 11.0 H 11.9 11.3 11.8 11.6 10.9 11.3 12.0 11.8 PHOS mg/dL 12.9 H 9.2 H 9.8 9.6 11.1 8.4 9.0 10.8 10.2 11.3 11.5 0.38 10 10.2 1.33 10 TRIG mg/dL MA mmt/l K Immol/L 7H 18 H 2 17 21 17 23 16 16 31 142 H 144 H 147 146 147 148 148 143 147 148 10.7 H 6.6 H 5.5 6.8 7.1 6.0 5.5 7.9 7.1 5.8 17 146 6.9 8.0 2.2 1.55 10 10 10 (--) - Data UnavaiLabLe LABCAT CC4.43 H - ltemolyzed 27-JUN-200_ 418-028:PAGE K-116 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT PERIOD: TERMINAL BY GROUP STUDY ID: ARGUS 418-O28 STUDY NO: 060-069 SEX: FERALE Animal TD GROUP: 1-F 19010 19012 19019 19021 19023 19041 19042 19044 19050 19053 19065 19068 19072 19074 19075 ALT AST U/L U/L ........ 144 123 136 182 177 167 115 114 142 154 104 121 153 149 156 157 87 123 80 133 ........ ......... ........ ......... ALP CA PHOS TRIG NA K U/L mg/clL mg/dL mg/dL mmot/l mmot/L 217 10.5 6.8 77 10.9 11.4 345 11.8 8.3 89 10.3 8.6 171 9.9 9.2 82 10.2 8.7 100 11.7 8.3 202 10.5 10.4 92 9.6 7.6 73 10.1 8.2 70 143 6.0 65 142 5.8 47 159 5.4 91 1/,O 5.5 46 140 6.2 44 138 5.7 70 149 4.8 51 140 7.4 40 140 5.7 20 143 5.6 MEAN SD N 129 31.7 10 142 22.8 10 145 88.7 10 10.6 0.72 10 8.8 1.33 10 54 19.9 10 143 5.8 6.3 0.67 10 10 (--) - Data Unavsi LabLe LABCAT CC4.43 27-JUN-2002 Study Report for Clinical INDIVIDUAL ANIMAL REPORT PERIOD = TERMINAL Chemistry BY GROUP 418-028:PAGE K,-117 STUDYIO: ARGUS418-028 STUDYNO: 060-069 Animal ID ALT AST UIL UIL GROUP:2-F 19004 19009 19016 19018 19026 19036 19037 19043 19047 19048 19052 19055 19061 19067 19071 97 133 116 110 150 141 136 102 145 142 176 125 115 181 150 139 117 131 132 147 .......... ........ ........ ......... ........ MEAN SD N 133 23.0 10 135 21.6 10 ALP U/L 80 193 242 240 183 200 48 252 46 102 CA _Iclk 11.3 11.7 11.6 11.6 10.7 11.9 10.9 11.5 10.0 10.4 PHOS mglclL TRIG _/dL 8.& 45 7.6 60 7.8 68 6.7 45 9.6 45 8.5 /,7 5.3 43 8.3 55 5.3 55 11.8 32 SEX: FEI4ALE NA mollL K motlL 139 5.3 144 5.9 147 5.4 144 5.0 141 5.0 145 5.4 143 5.2 142 6.1 142 5.6 140 6.9 159 81.7 10 11.2 0.63 10 7.9 1.94 10 50 10.2 10 143 5.6 2.4 0.58 10 10 (--) - Data Unavailable LABCATCC4./_3 2?-JUN-L_02 418-028:PAGE K-118 Study Report for Clinical Chemistry INDMDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060069 SEX: FEMALE Animal ID GROUP: 3-F 19003 19007 19008 19013 19014 19015 19017 1902/, 19029 19034 19038 19056 19057 19060 19064 MEAN SD N ALT AST U/L U/L 182 134 90 113 107 143 137 125 .......... 178 137 166 153 121 139 189 151 124 1&O ........ 130 133 ......... ......... ........ 142 34.2 10 137 11.8 10 ALP U/L 101 60 64 154 97 418 60 190 108 111 136 107.3 10 CA mg/dL 9.6 9.3 9.7 10.6 9.3 11.3 11.3 11.5 11.0 10.7 PHOS mg/dL 9.7 9.2 7.3 5.6 14.3 5-9 11.5 6.1 10.5 9.9 TRIG mg/dL 26 17 28 65 116 33 46 53 72 34 NA mmoL/L K mmot/L 142 5.5 139 5.8 139 5.7 144 5.4 141 4.9 145 5.4 142 6.0 147 5.4 161 5.5 1/.,2 5.9 10.4 0.87 10 9.0 2.80 10 49 29.4 10 142 5.6 2.5 0.32 10 10 (--) - Data Unavai LabLe LABC:AT CC4.43 27-JUN-2002 418-028:PAGE K-119 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDYID: ARGUS418-028 STUDYNO: 060-069 Animal ID GROUP:4-F 19002 19005 19035 19039 19040 19045 19046 19051 19054 19058 1-9062 19063 19066 19069 19073 &LT AST U/L U/L .......... 131, 127 115 128 174 125 134 132 157 149 ........ ........ 94 107 124 117 98 129 119 114 116 112 ........ ........ REAN SD N 127 24.7 10 12/* 12.1 10 ALP U/L 108 107 228 90 432 70 171 102 388 88 178 131.0 10 CA nlg/dL 10.6 10.0 11.0 9.7 11.1 10.0 10.7 10.3 10.7 10.3 10.4 0.46 10 PHOS Iig/dL TRIG IKj/dL SEX: FERALE NA _ot/L K mot/L 9.6 9.2 4.4 10.8 8.6 10.1 5.4 10.7 5.0 13.2 49 142. 6.2 67 142 5.5 62 147 5.8 77 138 7.0 55 145 5.9 28 139 6.5 30 142 5.3 18 144 7.1 34 143 5.1 33 142 6.6 8.7 2.88 10 45 19.5 10 142 6.1 2.6 0.70 10 10 (--) - Data UnavaiLabLe LABCATcc4.43 27-JUN-2002 418-028:PAGE K-120 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL STUDYI0: AR6US418-028 STUDYNO: 060--069 &nimal ID GROUP:5-F 19001 19006 19011 19020 19022 19025 19027 19028 19030 19031 19032 19033 190/,9 19059 19070 ALT AST U/L U/L 116 133 160 149 132 131 151 263 143 167 113 141 149 138 179 127 156 177 148 158 ........ ........ ......... ........ ......... MEAN SD N 145 20.0 10 156 40.7 10 ALP U/L 135 71 131 138 101 142 58 122 171 211 SEX: FERI_LE CA mcJ/dL 10.1 10.1 10.3 9.8 10.3 9.6 9.6 12.0 10.9 11.6 PHOS mg/dL 10.7 9.0 8.2 13.7 10.0 9.4 10.6 9.2 11.6 8.6 TRIG mmg/dL NA mmot/L 37 141 49 138 53 142 7 143 58 145 30 142 44 141 45 147 77 139 57 143 K mmot/L 6.8 6.8 5.2 6.9 6.4 5.4 7.6 5.1 6.3 5.1 128 44.7 10 10.4 0.82 10 10.1 1.64 10 44 18.3 10 142 6.2 2.6 0.90 10 10 (--) - Data UnavaiLabte LA_AT CC4.43 27- JUN-2002 STUDY1D: ARGUS418-028 STUDYNO: 060-069 418-028:PAGE K-121 study Report for Clinical INDIVIDUAL ANIMAL PERIOD: REPORT TERMINAL Chemistry BY GROUP SEX: MALE AniBa L IO CL Imot/L GROUP:1-M 19109 103 19115 100 19116 98 19119 100 19122 97 19125 96 19131 98 19132 99 19134 98 19135 93 19143 -- 19151 -- 19157 -- 19161 -- 19167 -- SEAS 98 SO 2.7 N 10 GtOB g/dL 2.0 2.1 1.8 1.9 1.9 2.2 2.3 1.8 2.0 2.3 -- ----- 2.0 0.19 10 A/G none 2.0 2.1 2.2 2.2 2.3 2.0 1.8 2.4 2.2 2.0 ------ 2.1 0.18 10 (--) - Data Unavailable I._BCATCC4._ 27- JUN-2002 STUDYID: ARGUS418-028 STUDYNO: 060-069 Study Report for Clinical INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL 418-028:PAGE K-122 .Chemistry BY GROUP SEX: MALE Animal ID 6ROUP: 2-M 19102 19106 19108 19110 19113 19120 19129 19136 19138 19139 19147 19153 19164 19165 19171 Ct mmt/L 98 99 101 101 98 96 99 99 99 98 ------ MEAN 99 SD 1.5 N 10 Gt._ g/dL 2.0 2.2 1.9 2.0 1.7 2.2 1.8 1.9 1.9 1.9 -- ----- 2.0 0.16 10 A/G none 2.1 1.8 2.1 2.1 2.3 2.0 2.2 2.3 2.3 2.2 ------ 2.1 0.16 10 (--) - Data UnavaiLabLe I.kBCAT CC4.43 27- JUN--2002 STUDYZD: ARGUS418-028 STUDYNO: 060-069 418-028:PAGE K-123 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL SEX: MALE Animal ID GROUP:3-M 19101 19105 19107 19112 19114 19121 19123 19130 19137 19146 19155 19159 19172 19173 19174 CL mmol./L 101 H 96 105 10(3 99 99 100 100 98 99 .... .... .... -.... MEAN 100 SD 2.3 N 10 GLOB g/dL 2.2 H 1.8 1.7 2.0 2.0 2.2 1.8 1.6 2.3 2.2 -- 2.0 0.24 10 A/G none 1.8 H 2.4 2.4 2.2 2.2 2.0 2.4 2.6 1.9 2.0 -- 2.2 0.26 10 (--) - Data Unavailable " LABCATCC4.43 H - Hemotyzed 27-JUN-2002 " STUDY ]D: ARGUS &1_ STUDY NO: 060-069 418-028:PAGE K-124 Study Report for Clinical INDIVIDUAL ANIMAL REPORT PERIOD : TERMINAL Chemistry BY GROUP SEX: HALE Animal ID GROUP: 4-M 19100 19103 19104 19118 19133 19141 19142 19144 191&B 19150 19156 19160 19162 19163 19166 CL mmmol/L 104 104 100 10.1 98 99 96 99 98 99 .... ----- REAN 100 SD 2.7 N 10 GLOB g/dL 2.0 1.7 1.8 1.8 1.9 1.9 2.2 2.1 2.1 2.0 ----- 2.0 0.16 10 A/G none 2.0 2.4 2.3 2.3 2.3 2.3 2.0 2.0 2.1 2.2 ----- 2.2 0.15 10 (--) - Data Unavai table LABCAT CC4.43 27- JtJN-2002 STUDY ID: ARGUS 418-028 STUDY NO: 060-069 418-028:PAGE K-125 Study Report for Clinical Chemistry INDMDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL SEX: M,_LE Animal ID GROUP: 5-M 19111 19117 19124 19126 19127 19128 19140 19145 19149 19152 19154 19158 19168 19169 19170 NEAN SD N CL mloL/C 101 H 101 H 99 100 101 98 101 98 100 99 -.... ---- 100 1.2 10 GLOB g/dL 2.0 H 1.8 H 1.8 1.8 2.0 1.7 1.7 1.9 1.8 1.6 -- ---- 1.8 0.13 10 A/G none 2.3 H 2.4 H 2.7 2.6 2.3 2.8 2.5 2.4 2.5 2.6 -- ---- 2.5 0.17 10 (--) - Data UnavailabLe LABCAT CC4.43 H - Hemolyzed 27-JUN-2002 STUDYID: ARGUS418-028 STUDYNO: 060-069 418-028:PAGE K-126 study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL SEX: FEI_ALE Animal ID GROUP:1-F 19010 19012 19019 10021 19023 19041 19042 19(0 19050 19053 19065 19068 19072 19074 19O75 MEAN SO N CL moL/L -96 92 100 91 93 92 96 99 99 98 ----- 96 3.4 10 SLOB g/dL -1.7 2.0 2.2 1.8 1.6 2.0 2.2 1.9 1.8 2.2 -- ---- 1.9 0.7.2 10 A/G none 2.2 Z.2 2.2 2.4 2.7 2.Z 2.0 2.1 2.1 1.6 --- --- 2.2 0.28 10 (--) - I)ata Unavailable LABCATCC4.43 27-JIJN-2(X]2 STUDYID: ARGUS418-028 STUDYNO: 060-069 418-028:PAGE K-127 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD: TERMINAL SEX: FERALE lUri_, L IO GROUP:2-F 19004 19009 19016 19018 19026 19036 19057 19O43 19047 19048 19052 19055 19O61 19067 1907'I MEAN SO N CL msol/L 89 93 98 97 94 95 96 91 93 95 ------ 94 2.7 10 GLOB g/dL 1.6 1.8 1.3 1.4 1.6 1.7 1.5 1.9 1.0 1.7 ------ 1.6 0.26 10 A/G none 2.7 2.2 3.2 3.0 2.4 2.5 2.3 2.0 3.3 2.5 ---- -- 2.6 0.43 10 (--) - Data UnavaiLable LABCATCC4.43 27-JUN-2002 STUDYI0: ARGUS418-028 STUDYNO: 060-069 , study Report for Clinical ZNDMDUAL _ PERIOD REPORT : TERMINAL Chemistry BY GROUP 418-028:PAGE K- 128 SEX: FEI_LE Animal I0 GROUP:3-F 19003 19007 190O8 19013 19014 19015 19017 1902& 19029 19034 19038 19056 19057 19060 19064 NEAN SD N CL mot/L 92 97 96 97 .... 92 99 94 101 88 -93 ---- 95 3.8 10 GLOB g/dk 1.7 1.5 1.7 1.7 1.9 1.7 2.2 1.8 2.I -1.9 ---- 1.8 0.21 10 A/G none 2.3 2.5 2.4 2.4 1.9 2.4 2.0 2.2 2.0 -2.2 ---- 2.2 0.21 10 (--) - Data UnavaiLabLe LABCAT CC4.43 27- JULY-2002  STUDYID: ARGUS418-028 STUDYNO: 060-069 418-028:PAGE K-.129 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL SEX: FENALE Animal 1D GROUP:&-F 19002 19005 19035 I_39 19040 19045 19046 19051 19054 19058 19O62 19063 19066 19069 190T5 MEAN SO N CL mmol./L -90 96 101 91 97 --94 97 98 97 93 --- 95 3.4 10 GLOB g/dL -1.8 1.7 1.5 1.9 1.9 --1.8 1.7 1.6 1.6 1.8 --- 1.7 0.13 10 A/G none -2.4 2.4 2.7 2.1 1.9 --2.5 2.3 2.8 2.3 2.3 --- 2.4 0.26 10 (--) - Data Unavaitable LABCAT CC4.43 27- JUN-2002 STUDYID: ARGUS418-028 STUDYNO: 060-069 418-028:PAGE K-130 Study Report for Clinical Chemistry INDIVIDUAL ANIMAL REPORT BY GROUP PERIOD : TERMINAL SEX: FEMALE Animal ID GROUP:5-F 19001 19006 19011 19020 19022 19025 19027 19028 19050 19031 19032 19033 19049 19059 19070 I_EAN SD N CL mol/L 9"1 93 92 91 100 94 96 95 95 89 _ -.... .... -- 94 3.1 10 GL08 g/dL 1.5 1.7 1.5 2.1 1.7 2.1 1.7 1.9 1.6 1.9 --- -- 1.8 0.21 10 A/E none 2.7 2.4 2.9 1.7 2.4 1.8 2.2 2.1 2.5 2.3 -- -- 2.3 0.37 10 (--) - Data Unavailable LABCATCC4.43 27-JUN-2002 418-028:PAGE K-131 study Report for Clinical Chemistry SUMMARY REPORT PERIOD TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETTtS PROCEDURE SEX: MALE TEST(s) : UNZTS: Group: 1-H MEAN SD N TP ALB GLU CHOL T-BZL BUN CREAT CK g/dL g/dL mg/dL mg/dL mj/dL mg/dL _j/dL U/L 6.3 0.36 10 4.3 0.22 10 145 16.8 10 57 0.2 8.3 0.07 10 10 16 0.3 288 1.5 0.04 172.6 10 10 10 ALT U/L 42 6.0 10 Group: 2-H HEAN SD N Group: 3-M MEAN SO N 6.1 0.28 10 6.3 0.33 10 4.1 0.19 10 4.3 0.15 10 141 26.0 10 182 58.2 10 41.* 11.1 10 O. 1 0.05 10 46 11.6 10 0.1 0.06 10 16 0.3 246 0.8 0._ 111.5 10 10 10 16 0.3 239 1.8 0.06 112.9 10 10 10 63 69.9 10 60 34.4 10 Group: 4-I_ MEAN SD N Group: 5-M MEAN SD N 6.2 0.28 10 6.3 0.28 10 /+.2 0.17 10 162 25.2 10 4.5** 0.20 10 165 33.7 10 43* 13.1 10 0.1 0.05 10 33** 7.2 10 0.2 0.06 10 17 0.3 196 95 1.6 0.03 53.5 124.1 10 10 10 10 21"* 0.3 243 45 2.4 0.05 117.4 7.0 10 10 10 10 *-Significant Difference from Control P < .05 LABCAT CC4.43 **-Significant Difference from Control P < .01 27-JUtt-2002 418-028:PAGE K-132 study Report for Clinical Chemistry SUMMARY REPORT PERIOD : TERMINAL STUDY ID: ARGUS /18-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE TEST(s) : UNITS: Group: 1-H MEAN SD N AST U/L 96 22.4 10 ALP CA PHOS TRIG NA K CL U/L IIg/dL Ing/dL mg/dL mol/L mot/L mol/k 105 10.9 9.5 52 146 6.6 98 q4.3 0.48 1.40 21.8 1.4 2.10 2.7 10 10 10 10 10 10 10 SLOB g/dL 2.0 0.19 10 Group: 2-M MEAN SD N 121 92.6 10 111 36.9 10 10.7 0.44 10 9.4 1.11 10 47 17.2 10 146 6.1 1.3 0.89 10 10 99 2.0 1.5 0.16 10 10 Group: 3-H MEAN SO N 117 /,0.9 10 100 12.4 10 11.1 0.44 10 9.9 2.09 10 36 13.8 10 147 6.5 1.6 1.27 10 10 100 2.0 2.3 0.24 10 10 Group: 4-M MEAN SD N 198 248.7 10 115 25.1 10 11.1 0.34 10 9.7 1.1_3 10 56 27.9 10 147 6.0 1.0 0.60 10 10 100 2.0 2.7 0.16 10 10 Group: 5-M MEAN SD N 97 15.6 10 1/##nt 37.5 10 11.5** 0.38 10 10.2 1.33 10 17"* 146 6.9 8.0 2.2 1.55 10 10 10 100 1.8 1.2 0.13 10 10 **-Significant Difference from Control P < .01 LABCAT CC4.43 27-JUN-2002 STUDYID: ARGUS18-028 STUDYNO: 060-069 418-028:PAGE K-133 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD - TERMINAL ANALYSZSOF VARIANCEFOLLOWEDBY DUNNETT'SPROCEDURE SEX: MALE TEST(s) : UNZTS: A/6 none Group: 1-M MEAN SD N 2.1 0.18 10 6roup: MEAN SD N 2-11 2.1 0.16 10 Group: 3-M MEAN SD N 2.2 0.26 10 Group: 4-M MEAN SD N 2.2 0.15 10 6roup: 5-M MEAN SO N 2.5** 0.17 10 **-Significant Difference from Control P < .01 LABCATCC4./,3 27- JUN-2002 418-028:PAGE K- 134 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE Total Protein Group 1-M 2-H 3-H 4-H 5-11 N Total 10 62.8 10 60.6 10 62.5 10 61.8 10 63.3 Rean 6.3 6.1 6.3 6.2 6.3 Std. Dev. DUNNETT'S 't' O. 36 0.28 1.60 0.33 0.22 0.28 0.73 0.28 0.36 DUNNETT'S RANGES LO -95%- HI LO -99X- HI 5.9 6.6 5.9 6.6 5.9 6.6 5.9 6.6 5.9 6.7 5.9 6.7 5.9 6.7 5.9 6.7 Source Degree Fdm TREATMENTS 4 ERROR 45 TOTAL 49 F Ratio = 1.16 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 4.948 gumnett's 'T' table values P.01 : 3.12 P.05 = 2.51 Albumin 1-M 2_ 3-M 4-M 5-H 10 42.5 4.3 10 41.1 4.1 lo 42.7 4.3 10 42.3 4.2 10 45.2 4.5 0.22 0.19 0.15 0.17 0.20 1.66 0.24 0.24 3.2q 4.0 4.0 4.0 4.0 4.5 4.5 4.5 4.5* F Ratio = Coeff. Var. % = 6.35 4.399 'F' table Dunnett's vaLues 'T' table values F.01 = P.01 = Glucose 4.0 4.0 4.0 4.0 TREATMENTS 4 4.5 ERR 45 4.5 4.5 TOTAL 49 4.5** 3.78 F.05 = 3.12 P.05 -- 2.58 2.51 1-M 2-M ]-M 4-M 5-44 10 1453 10 1411 10 1823 10 1619 10 1649 145 141 182 16,?. 165 16.8 26.0 58.2 25.2 33.7 0.27 2.37 1.06 1.25 106 185 106 185 106 185 106 185 TREATHENTS 4 96 194 96 194 ERROR 45 96 1_ 96 19/+ TOTAL 49 F Ratio = 2.24 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 21.988 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 SEX: MALE Sum of Squares O. 44 4.26 4.70 Mean Square O. q 1 0.09 0.90 1.59 2.49 0.22 0.04 I_ 55071 06013 27"35 1224 *-Significant **-Significant Error-within Difference Difference groups from Control froB Control P < .05 P < .01 LABCAT CC4.43 Source-Source of Variation Treatments-between groups 27-JUN-2002 418-028:PAGE K- 135 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD : TERMINAL STUDY !D: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE Cholesterol Group 1-M 2-M 3-M 4-M 5-M N Total 10 569 10 413 10 459 10 427 10 331 Mean 57 41 46 43 33 Std. Dev. DUNNETT'S 't' 8.3 11.1 11.6 13.1 7.2 3.32 2.34 3.03 5.07 OUNNETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm TREATMENTS 4 45 69* 42 72** ERROR 45 45 69 42 72 45 69* 42 72 TOTAL 49 45 69* 42 72** F Ratio = 6.76 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vsr. % = 2_3.859 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Total Bilirubin 1-M 10 2-M 10 3-M 10 4-M 10 5-M 10 F Ratio = Coeff. Vat. % = 1.6 0.2 0.07 1.4 0.1 0.03 0.76 0.1 0.2 1.2 0.1 0.06 1.53 0.1 0.2 1.3 0.1 0.05 1.15 0.1 0.2 1.9 0.2 0.06 1.15 0.1 0.2 2.25 39.527 ' F' table Ounnett's values 'T' table values F.01 = P.01 = TREATMENTS 4 0.1 0.2 ER_R 45 0.1 0.2 0.1 0.2 TOTAL 49 0.1 0.2 3.78 F.05 = 2.58 3.12 P.05 = 2.51 Blood Urea Nitrogen 1-M 10 158 16 1.5 TREATMENTS 4 2-M 10 162 16 0.8 0.53 14 18 13 18 ERROR 45 3-. 10 162 16 1.8 o.s3 14 is 13 18 4-M 10 174 17 1.6 2.11 14 18 13 18 TOTAL 49 5-M 10 207 21 2.4 6.46 14 18. 13 18"* F Ratio = Coeff. Var. % = 14.12 9.821 'F' table Durmett's values 'T' table values F.01 = P.01 = 3.78 3.12 F.05 = P.O5 = 2.58 2.51 SEX: RALE Sum of Squares 2978.06 4954.90 7932.9B Mean Square 744.52 110.11 0.031 0.154 0.185 0.008 0.003 162.32 129.30 291.62 40.58 2.87 *-Significant /nt-Significant Error-within Difference Difference groups from Control from Control P < .05 P < .01 LABCAT CC4.43 Source-Source of Variation Treatments-between groups 27-JUN-2(X)2 418-028:PAGE K-136 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARZANCE FOLLOWEDBY DUNNETT'5 PROCEDURE Creatinine Group N Total 1lean Std. Dev. DUt_ETT' S 't' DUNNETT'S RANGES LO -95%- HZ LO -99%- H1 Source Degree Fdm 1-11 2-11 3-H 4-H 5-H '10 3.2 0.3 0.06 10 3.3 0.3 0.05 0.48 10 3.1 0.3 0.06 0.48 10 3.1 0.3 0.03 0.48 10 3.3 0.3 0.05 0.48 0.3 0.4 0.3 0.4 0.3 0.4 0.3 0.4 TREATHENTS 4 0.3 0.4 ERROR 45 0.3 0.4 0.3 0.4 TOTAL 49 0.3 0.4 F Ratio = 0.47 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 14.434 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Creatine Kinase 1-11 2-H 3-. &-11 5-11 10 2877 10 24,62 10 2388 10 1964 10 2427 288 246 239 196 243 172.6 111.5 112.9 53.5 117.4 0.78 0.91 1.71 0.84 153 153 153 153 422 422 422 422 121 121 121 121 TREATMENTS 4 455 455 ERROR 45 455 TOTAL 49 455 F Ratio : 0.73 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. VaT. % = 49.381 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Alanine Aminotrans ferase I-H 2-11 3-. 4-11 5-11 10 420 10 631 10 600 10 954 10 447 42 6.0 63 69.9 0.72 60 34.4 0.61 95 124.1 1 ._. 45 7- 0 O. 09 -32 116 -32 116 -32 116 -32 116 TREATIqENTS 4 -50 134 ERROR 45 -50 134 -50 134 TOTAL 49 -50 134 F Ratio = Coeff. VaT. X = 1.05 107.598 'F' table Dunnett's values 'T' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 SEX: 11ALE Sum of Squares O. 0040 0.0960 0.1000 1lean Square 0.0010 0.0021 41956 644538 686694 10489 14323 18155 194111 212266 4539 4314 Error-vithin groups Source-Source of Variation LABCAT CC4.43 Treatments-between groups 27-JUN-2002 418-028:PAGE K-137 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD : TERMINAL STUDY ID: ARGUS &IB-OZ8 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLONEDBY DUNNETT'5 PROCEDURE Aspartate Group N Total _minotra_s ferase Std. DUNNETT'S Rean Dev. 't ' DUNNETT'S RANGES LO -95%- HI LO -99Z- HI Source Degree Fdm 1-H 10 957 96 22.4 TREATMENTS 4 2-N 10 1213 121 92.6 0.47 -40 231 -73 264 ERROR 45 3-M 10 1171 117 40.9 0.40 -40 231 -73 264 4-R 10 1976 198 248.7 1.89 -40 231 -73 264 TOTAL 49 5-H 10 974 97 15.6 O. 03 -40 231 -73 264 F Ratio = 1.20 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 95.916 Puonett's 'T' table values P.01 = 3.12 P.05 = 2.51 Alkaline Phosphatase 1-R 10 1050 105 14.3 2-H 10 1113 111 36.9 0.51 74 136 3-. 10 lOre lOO 12.4 0.6o 74 136 4-R 10 1152 115 25.1 0.83 74 136 5-H 10 1441 t41+ 37.5 3.19 74 136. f Ratio = Coeff. Vat. % = 3.94 23.795 'F' table Dunnett's values 'T' table values F.01 = P.01 = Calcium TREATMENTS 4 67 143 67 143 ERROR 45 67 143 TOTAL 49 67 143.* 3.78 F.05 = 2.58 3.12 P.05 = 2.51 1-R 10 109.4 10.9 0.48 TREATMENTS 4 2-M 10 107.3 10.7 O.Z_; 1.12 10.5 11.4 10.4 11.5 ERROR 45 3-. 10 110.7 11.1 0._ 0.69 10.5 11.4 _0.4 11.5 4-H 10 111.0 11.1 0.34 0.85 10.5 11.4 10.4 11.5 TOTAL 49 5-H 10 115.3 11.5 0.38 3.14 10.5 11.4* 10.4 11.5** F Ratio = Coeff. Vat. Z = 4.87 3.800 'F' table Dunnett's values 'T' table values F.01 = P.01 = 3.78 3.12 F.05 = P.05 = 2.5B 2.51 SEX: NALE Sum of Squares 69637 655378 725015 Mean Square 17/+09 14564 11825 33780 751 45604 3.45 0.86 7.97 0.18 11.42 *-Significant **-Significant Error-within Difference Differone groups fro= Control frmm Control P < .05 P < .01 _CAT CC4.43 Source-source of Variation Treetaents-between groups 27- JUN-2O02 418-028:PAGE K-138 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY lO: ARGUS 418-028 STUDY NO: O60-069 ANALYSIS OF VARXANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: MALE Phosphorus Group N Total Mean Std. Dev. DUNNETT'S 't' DUNNETT' S RANEES LO -95%- HI LO-99%- HI Source Degree Fdm 1-M 10 94.7 9.5 1.40 TREATMENTS 4 2-M 10 93.7 9.4 1.11 0.15 7.8 11.2 7.4 11.6 ERROR 45 3-M 10 99.1 9.9 2.09 0.65 7.8 11.2 7.4 11.6 4-H 10 97.1 9.7 1.43 0.36 7.8 11.2 7.4 11.6 TOTAL 49 5-M 10 102.3 10.2 1.33 1.13 7.8 11.2 7.4 11.6 F Ratio = 0.53 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. Z : 15.485 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Triglycerides 1-M 10 516 52 21.8 2-tt 10 469 47 17.2 0.55 3-. 10 364 36 13.8 1.z9 4-M 10 361 36 27.9 1 .B2 5-M 10 168 17 B.O 4.10 30 73 3o 73 30 73 30 73* TREATMENTS 4 25 78 25 78 ERROR 45 25 78 TOTAL 49 25 78** F Ratio = 4.98 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 50.584 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sodiu= 1-14 10 1459 146 1.4 TREATMENTS 4 2-H 10 1457 146 1.3 0.20 1_ 168 144 168 ERROR 45 3-. 1o 1tin5 147 1.6 o._ I_ 1_ I_ I_ 4-ti 10 1469 147 1.0 1.44 1_ 148 1/,4 1/,8 TOTAL 49 5-M 10 1460 1/+6 2.2 0.14 144 148 144 148 F Ratio = Coeff. Vat. _ = 1.00 1.062 'F' table Dunnett's values 'T' table values F.01 = P.01 = 3.78 L05 = 3.12 P.05 = 2.58 2.51 Sum of Squares 4.80 102.32 107.12 7188 162_; 9.60 108.40 118.00 Mean Square 1.20 2.27 1797 361 2.40 2.41 *-Significant _rk-Significant Error-within Difference Difference groups from Control fro= Control P < .05 P < .01 LABCAT CC4.43 Source-Source of TreatNnts-betveen Variation groups 27- JUfl-2002 418-028:PAGE K-139 " Study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 Potassium Group 1-H 2-H 3-g 4.-H 5-H N Total 10 65.7 10 61.0 10 64.5 10 59.7 10 69.0 Hean 6.6 6.1 6.5 6.0 6.9 ANALYSIS OF VARIAHCE FOLLOWEDBY DUNNETT'S PROCEDURE Std. Dev. DUNNETT'S 't' DUNNETT'S RANGES LO -95%- HI LO -99H- HI Source Degree Fdm 2.10 0.89 0.76 1.27 0.19 0.60 0.97 1.55 0.53 5.0 8.1 5.0 6.1 5.0 8.1 5.0 8.1 TREATHENTS 4 4.6 8.5 ERROR 4.5 4.6 8.5 4.6 8.5 TOTAL 49 4.6 8.5 F Ratio = 0.73 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. X = 21.633 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Chloride 1-H 10 982 98 2.7 2-. 10 9era 99 1.5 0.62 96 101 3-. 10 997 100 2.3 1.54 96 lol 4-H 10 1000 100 2.7 1.85 96 101 5-H 10 998 100 1.2 1.64 96 101 F Ratio = Coeff. Var. % = 1.25 2.190 'F' table Dunnett's values 'T' table values F.01 = P.01 = Globulin TREATRENTS 4 95 lm ERROR45 9s lm 95 101 TOTAL 49 95 101 3.78 F.05 = 2.58 3.12 P.05 = 2.51 I-R 2-11 3-11 4-R 5-N 10 20.3 2.0 0.19 10 19.5 2.0 0.16 0.99 10 19.8 2.0 0.24 0.62 10 19.5 2.0 0.16 0.99 10 1B.I 1.8 0.13 2.73 1.8 2.2 1.8 2.2 1.8 2.2 1.8 2.2* TREATRENTS 4 1.8 2.3 1.8 2.3 ERROR 45 1.8 2.3 1.8 2.3 TOTAL 49 F Ratio = Coeff. Vat. % = 2.06 9.253 'F' table Dunnett=s values 'T' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 SEX: HALE Sum of Squares 5.56 86.21 91.77 H_n SqUare 1.39 1.92 23.60 m2.90 236.50 5.90 4.73 0.27 0.07 1.46 0.Q3 1.72 *-Significant Error-within Difference groups from Control P < .05 LABCAT CC4.43 Source-Source of Varietion Treatments-between groups 27-JUN-2002 418-028:PAGE K- 140 Study Report for Clinical SUMMARY REPORT PERIOD: TERMINAL Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSZS OF VARIANCE FOLLORED BY DUNNETT'S PROCEDURE Albumin/Globulin Group N Total Mean Ratio $td. DUNNETT'S Per. 't' DUNNETT'S RANGES LO-95_{- HI LO -997..- HI Source Degree Fda 1-M 10 21.2 2.1 0.18 TREATMENTS 4 2-H 10 21.4 2.1 0.16 0.24 1.9 2.3 1.9 2.& ERROR 45 3-M 10 21.9 2.2 0.26 0.84 1.9 2.3 1.9 2.4 4-M 10 21.9 2.2 0.15 0.84 1.9 2.5 1.9 2.4 TOTAL 69 5-N 10 25.1 2.5 0.17 4.67 1.9 2.3* 1.9 2.6/:* coeff. F Ratio = Van. % = 7.31 8.368 'F' 1:able values Dunnetl:'s 'T' table values F.01 = P.01 = 3.78 F.05 = 3.12 P.05 = 2.58 2.51 SEX: MALE Sum of Squares 1.02 1.57 2.59 Mean Square 0.25 0.03 *-Significant **-Significant Error-within Oiffevence Difference groups from Control from Control P < .05 P < .(_1 L_JBCAT CC4.43 Source-Source of Variation Treatments-between groups 27- JUN-2(X)2 418-028:PAGE K-141 Study Report for Clinical Chemistry SUMMARY PERIOD: REPORT TERMINAL STUDY I0: ARGUS 418-028 STUDY NO: 060-069 ANALYSZS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: FEMALE TEST(S) : UNITS: Group: 1-F MEAN SD N Group: 2-F MEAN SD H Group: 3-F MEAN SD N TP AIR 6LU CHOL T-BIL BUN CREAT CK ALT g/dL g/dL mg/dL mg/dL ug/dL mg/dL mg/dL U/L U/L 6.1 0.52 10 4.1 0.42 10 152 19.8 10 77 27.8 10 0.2 0.07 10 28 O. 4 202 129 4.4 0.05 54.1 31.7 10 10 10 10 5.5 0.50 10 4.0 0.34 10 162 20.0 10 72 14.0 10 O. 2 0.03 10 24 O.4 250 133 4.7 0.05 85.6 23.0 10 10 10 10 5.9 0.41 10 4.1 0.24 10 155 18.7 10 71 20.0 10 0.2 0.05 10 28 0.3 192 142 6.7 0.05 21.7 34.2 10 10 10 10 Group: 4-F MEAN SD N 5.8 0.32 10 4.0 0.29 10 146 16.2 10 02 14.2 10 0.1 0.05 10 25 0.4 213 127 &.4 0.1)8 80.4 24.7 10 10 10 10 Group: 5-F MEAN SD N 5. B 0.29 10 4.0 0.31 10 140 35.5 10 61 19.8 10 O.I 0.05 10 30 15.4 10 O. 3 0.06 10 252 101.6 10 145 20.0 10 LABCAT CC4.43 27-JUN-2002 418-028:PAGE K-142 Study Report for Clinical Chemistry SUMMARY PERIOD: REPORT TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 TEST(s) : UNITS: Group: 1-F MEAN SD N Group: 2-F MEAN SD N AST U/L 142 22.8 10 135 21.6 10 ANALYSZS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: FEMALE ALP CA PHOS TRIG NA K CL U/L mg/dL mg/dL mg/dL mmt/L mmL/L uot/L GLOB g/dL 145 88.7 10 10.6 0.72 10 8.8 1.33 10 54 19.9 10 143 5.8 6.3 0.67 10 10 96 1.9 3.6 0.22 10 10 159 81.7 10 11.2 0.63 10 7.9 1.96 10 50 10.2 10 143 5.6 2.4 0.58 10 10 94 1,6** 2.7 0.26 10 10 Group: 3-F MEAN SD N Group: 4-F MEAN SD N Group: 5-F MEAN SD N 137 11.8 10 136 107.3 10 124 12.1 10 178 131.0 10 156 40.7 10 128 44.7 10 10.4 0.B7 10 10.4 0.46 10 10.4 0.82 10 9.0 2.80 10 8.7 2.88 10 10.1 1.64 10 49 29.4 10 45 19.5 10 44 18.3 10 142 5.6 2.5 0.32 10 10 142 6.1 2.6 0.70 10 10 142 6.2 2.6 0.90 10 10 95 1.8 3.8 0.21 10 10 95 1.7 3.4 0.13 10 10 94" 3.1 10 1.8 0.21 10 _rk-Significant Difference from Control P < .01 LABCAT CC6.43 27- JUN-2002 STUDYID: AREUS418-028 STUDYNO: 060-069 418-028:PAGE K-143 Study Report for Clinical SUMMARY PERZOD: REPORT TERMINAL Chemistry ANALYSISOF VARIANCEFOLLOWEBDY DUNNETT'SPROCEDURE SEX: FEMALE TEST(s): UNITS: Group: 1-F mEAN SD N 6roup: 2-F MEAN SO N Group: 3-F NEAN SD N Group: 4-F HFAN SD I_ Group: 5-F MEAN SO N A/G none 2.2 0.28 10 2.6* 0._ 10 2.Z 0.21 10 2.4 0.26 10 2.3 0.37 10 *oSignificant Difference from Control P < .05 LABCATCC4.43 27-JUN-L_02 418-028:PAGE K-144 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE Total Protein Group 1- F 2-F 3-F 4-F 5-F N Total 10 60.7 10 55.0 10 58.8 10 57.7 10 58.2 Mean 6.1 5.5 5.9 5.8 5.8 Std. Dev. DUNNETT'S 't' O. 52 0.50 0.41 0.32 0.29 3.1)4 1.01 1.60 1.33 DUNNETT'S RANGES LO -95Z- HI LO -99%- HI Source Degree Fdlm 5.6 6.5* 5.6 6.5 5.6 6.5 5.6 6.5 TREATMENTS & 5.5 6.7 ERROR 45 5.5 6.7 5.5 6.7 TOTAL 69 5.5 6.7 F Ratio = 2.42 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 7.220 Dunnett's 'T' table values P.01 = 3.12 P.05 : 2.51 2_bua_n 1-F 2-F _P'-F 4-F 5-F 10 41.3 4.1 0.42 10 39.5 A.O 0.34 1.24 10 40.6 4.1 0.24 0.48 10 40.4 4.0 0.29 0.62 10 40.3 4.0 0.31 0.69 3.8 4.5 3.8 4.5 3.8 4.5 3.8 4.5 TREATMENTS 4 3.7 4.6 ERROR 45 3.7 4.6 3.7 4.6 TOTAL 49 3.7 4.6 F Ratio = 0.39 'F' table values F.01 = 3.78 F.05 2.58 Coeff. Var. % = 8.059 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Glucose 1-F 10 1520 152 19.8 TREATHENTS 4 2-F 10 1621 162 20.0 0.98 126 178 120 184 ERROR 45 ]-F 10 1548 155 18.7 0.27 126 178 120 184 4-F 10 1401 146 16.2 0.57 126 178 120 184 TOTAL 49 5-F 10 1404 140 35.5 1.12 126 178 120 184 F Ratio = 1.29 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 15.288 Durmett's 'T' table values P.01 = 3.12 P.05 = 2.51 SEX: FEMALE Sum of Squares 1.70 7.9'1 9.62. Mean Square O. 43 0.18 0.17' 4.78 4.94 0.04 0.11 2750 687 24006 533 26756 *-Significant Error-within Difference groups from Control P < .05 LABCAT cc4.43 Source-Source of Variation Treacmnts-betueen groups 27- JUN-2002 418-028:PAGE K-145 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE Cholesterol Group 1-F 2-F 3-F 4-F 5-F N Total 10 769 10 723 10 705 10 621 10 610 Mean 77 72 71 62 61 Std. Oev. DUNNETT'S 't ' DUNNETT'S RANGES LO -95%- HI LO -99%- HZ Source Pegree Fdm 27.8 14.0 20.0 14.2 19.8 0.52 0.72 1.67 1.79 55 99 55 99 55 99 55 99 TREATMENTS & 49 105 ERROR 45 49 105 49 105 TOTAL 49 49 105 F Ratio : 1.18 'F' table values F.01 = 3.?8 F.05 = 2.58 coeff. VaT. % = 28.929 Durmettts 'T' table values P.01 = 3.12 P.05 = 2.51 Total Biliz_zbin 1-F 2-F 3.-F 4-F 5-F 10 1.6 0.2 0.07 10 1.9 0.2 0.03 1.30 0.1 O.Z 10 1.5 0.2 0.05 0._ 0.1 0.2 10 1.3 0.1 0.05 1.30 0.1 0.2 10 1.3 0.1 0.05 1.30 0.1 0.2 F Ratio = Coeff. VaT. % = 2.33 33.974 'F' table Dunnett's values 'T' table values F.Oq = P.01 = Blood Urea Nitrogen 1- F 10 281 28 4.4 2-F 10 241 24 4.7 1.08 19 37 3-F 10 281 28 6.7 0.00 19 37 4-F 10 252 25 4.4 0.78 19 37 5-F 10 296 30 15.4 0.46 19 37 F Ratio = Coeff. Var. % = 0.79 30.672 'F i table Dunnett's values 'T' table values F.01 = P.01 = TREATMENTS 4 0.1 0.2 0.1 0.2 ERROR45 0.1 0.2 TOTAL 49 0.1 0.2 3.78 F.05 = 2.58 3.12 P.05 = 2.51 TREATMENTS 4 17 40 17 40 ERROR 45 17 40 TOTAL 49 17 40 3.78 F.05 = 3.12 P.05 = 2.58 2.51 SEX: FEMALE Sum of Squares 1862 17702 19564 Mean Square /,65 393 0.025 0.120 0.145 0.006 0.003 218.92 3099.90 3318.82 54.73 68.89 Error-within groups Source-Source of Variation LABCAT CC4.43 Treatmenta-betu_-en groups 27-JUN-2002 418-028:PAGE K-146 Study Report for Clinical SUMMARY REPORT PERIOD : TERMINAL Chemistry STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE Creatinine Group 1-F 2-F 3-F 4-F 5-F N Total 10 3.5 10 3.5 10 3.4 10 3.6 10 3.1 Mean 0.4 0.4 0.3 0.4 0.3 Std. Oev. DUNNETT'S 't' 0.05 0.05 0.05 0.08 0.06 0.00 0.37 0.37 1.47 DUNNETT'S RANGES LO -95%- HI LO -99_- HI 0.3 0.4 0.3 0.4 0.3 O.& 0.3 0.4 0.3 0.4 0.3 0.4 0.3 0.4 0.3 0.4 Source Degree Fdm TREATMENTS 4 ERROR 45 TOTAL 49 F Ratio = 1.00 'F' table values F.01 = 3.78 F.O5 = 2.58 Coeff. Var. % = 17.813 Dunnett's 'T' table values P.01 = 3.12 P.O5 = 2.51 Creatine Kinase I-F 2-F 3-r 4-F 5-F 10 2017 10 2495 10 1919 10 2129 10 2521 202 250 192 213 252 54.1 85.6 21.7 80.4 101.6 1.44 0.30 0.34 1.52 118 118 118 118 285 285 285 285 TREATMENTS 4 9B 305 98 ERROR 45 98 305 TOTAL 49 98 305 F Ratio = 1.39 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 33.472 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Alanine Aminotrans ferase SEX: FEMALE Sum of Squares 0.015 0.167 0.182 Mean Square 0.004 0.004 30624 247619 278244 7656 5503 1-F 10 1294 129 31.7 TREATMENTS 4 2546 637 2-F 10 1334 133 23.0 0.33 99 160 91 167 ERROR 45 33396 742 3-F 10 1424 142 34.2 1.07 99 160 91 167 4-F 10 1265 127 24.7 0.24 99 160 91 167 TOTAL 49 35942 5-F 10 1447 145 20.0 1.26 99 160 91 167 F Ratio = 0.86 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. Z = 20.138 Dunnett_s 'T' table values P.01 = 3.12 P.05 = 2.51 Error-within groups _)urce-$_)urce of Variation _CAT CC4.43 Treatments-between gr_s PT-JUN-_ 418-028:PAGE K_147 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD: TERMINAL STUDY I0: ARGUS 418-028 STUDY NO: 060-069 ANALYSZS OF VARIANCE FOLLOUEDBY DUNNETT'S PROCEDURE SEX: FEMALE Aspartate Group N Total Aminotrans Std. Mean Dev. ferase DUNNETT'S 't' DUNNETT' S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm 1-F 10 1423 142 22.8 TREATMENTS 4 2-F 10 1351 135 21.6 0.67 115 169 109 176 ERROR 45 3-F 10 1368 137 11.8 0.51 115 169 109 176 4-F 10 1240 124 12.1 1.69 115 169 109 176 TOTAL 49 5-F 10 1564 156 40.7 1.30 115 169 109 176 F Ratio = Coeff. Var. % = 238 17.422 'F' table Dunnett's values 'T' table values F.01 = P.Oq = Alkaline Phosphatase 1-F 10 1/.d_ 145 88.7 2-F 10 1586 159 81.7 0.32 38 252 3-F 10 1363 136 107.3 0.20 38 252 4-f 10 1784 178 131.0 0.79 38 252 5-F 10 1280 128 /_.7 0.40 38 252 3.78 F.OS = 3.12 P.05 = 2.58 2.51 TREATMENTS & 12 277 ERROR 45 12 277 12 277 TOTAL 49 12 277 F Ratio = Coeff. Vat. % = 0.44 63.7_3 'F' table Dunnett's values 'T' table values F.01 = P.01 = Calcium 1-_ 10 105.5 10.6 0.72 2-F 10 111.6 11.2 0.63 1.90 9.7 11.4 3-F 10 104.3 10.4 0.87 0.37 9.7 11.4 4-F 10 104.4 10.4 0.46 0.3/+ 9.7 11.4 5-F 10 104.3 10.4 0.82 0.37 9.7 11.4 3.78 F.05 = 2.58 3.12 p.05 = 2.51 T_T_E.TS 4 9.5 11.6 ERROR 45 9.5 11.6 9.5 11.6 TOTAL 49 9.5 11.6 F Ratio = 1.94 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 6.760 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares 5587 26359 31946 Rean Square 1397 586 15762 406995 422757 3941 9044 3.99 23.11 27.11 1.00 0.51 Error-within groups Source--Source of Variatio_ "LABCAT CC4.43 Treatl_nts-between groups 27-JUN-2002 418-028:PAGE K-148 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD : TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUflNETT'S PROCEDURE SEX: FEMALE Phosphorus Group N Total Mean Sl:d. Dev. OUNHETT' S ' t' DUNHETT'S RANGES LO -_%- HI LO -99%- HI Source Degree Fd_ 1-F 10 87.5 8.8 1.33 TREATMENTS 4 2-F 10 79.3 7.9 1.94 0.83 6.3 11.2 5.7 11.8 ERROR 45 3-F 10 90.0 9.0 2.80 0.25 6.3 11.2 5.7 11 .B 4-F 10 87.0 8.7 2.88 0.05 6.3 11.2 5.7 11.8 TOTAL 49 5-F 10 101.0 10.1 1.64 1.37 6.3 11 .Z 5.7 11.8 F Ratio = 1.26 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Var. % = 24.803 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares 24.53 219.09 243.62 Mean Square 6.13 4.87 Triglycerides q-F 10 544 54 2-F 10 495 50 3-F 10 490 49 4-F 10 453 45 5-F 10 437 44 19.9 10.2 0.54 29.4 0.59 19.5 1.00 18.3 1.17 32. 77 32 77 32 77 32 77 F Ratio = Coeff. Var. % = 0.42 42.148 'F' table Du_nett_s values 'T' table values F.01 = P.01 = Sodium 1-F 10 1434 143 6.3 2-F 10 1427 143 2.4 0.43 139" 147 3-F 10 1422 142 2.5 0.74 139 147 4-F 10 1424 142 2.6 0.62 139 147 5-F 10 1421 142 2.6 0.80 139 147 F Ratio = Coeff. Vat. % = 0.21 2.554 'F' table Dunnett's values 'T' table values F.01 = P.01 = TREATRENT$ 4 26 83 ERROR 45 26 83 26 83 TOTAL 49 26 83 3.78 F.05 = 2.58 3.12 P.05 = 2.51 TREATHENTS 4 138 148 ERROR 45 138 148 138 148 TOTAL 49 138 148 3.78 F.05 = 2.58 3.12 P.05 = 2.51 693 18711 19404 10.92 587.40 596.32 173 416 2.73 13.05 Error-within groups Source-Source of Variation LABCAT CC4.43 Treatlnts-betweenoroUl)S 27-JUN-2002 418 -028 :PAGE K- 149 I Study Report for Clinical Chemistry SUMMARY REPORT PERIOD : TERMINAL STUDY ID: ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOWEDBY DUNNETT'S PROCEDURE SEX: FEMALE Potassium Group N Total Mean Std. Dev. DUNNETT'S 't' DUNNETT'S RANGES LO -95%- HI LO -99%- HI Source Degree Fdm 1-F 10 58.1 5.8 O. 67 TREATMENTS 4 2-F 10 55.8 5.6 0.58 0.78 5.1 6.6 4.9 6.7 ERROR 45 3-F 10 55.5 5.6 0.32 0.88 5.1 6.6 4.9 6.7 4-F 10 61.0 6.1 0.70 0.98 5.1 6.6 4.9 6.7 TOTAL 49 5-F 10 61.6 6.2 0.90 1.18 5.1 6.6 4.9 6.7 F Ratio : Coeff. Van. % = 1.8& 11.322 Chloride 'F' table Dunnett's values 'T' table values F.01 : P.01 : 3.78 F.O5 = 2.58 3.12 P.05 : 2.51 1- F 10 956 96 3.4 TREATMENTS 4 2-F 10 941 94 2.7 1.01 92 99 91 100 ERROR 4.5 }-F 10 949 95 3.8 0.47 92 99 91 100 4-F 10 954 95 3.4 0.14 92 99 91 100 TOTAL 49 5-F 10 936 94 3.1 1.35 92 99 91 100 F Ratio = 0.66 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. VaT. % = 3.494 Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Globulin 1-F 10 19.4 1.9 0.22 TREATMENTS 4 2-P 10 15.5 1.6 0.26 4.12 1.7 2.2* 1.6 2.2** ERROR 45 3-F 10 18.2 1.8 0.21 1.27 1.7 2.2 1.6 2.2 4-F 10 17.3 1.7 0.13 2.22 1.7 2.2 1.6 2.2 TOTAL &9 5-F 10 17.9 1.8 0.21 1.59 1.7 2.2 1.6 2.2 F Ratio = 4.56 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. VaT. % = 11.982 Ounnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares 3.23 19.67 22.90 29.08 493.00 522.08 0.82 2.02 2.83 Mean Square O. 81 0.44 7.27 10.96 0.20 0.04 *-Significant *k-Significant Error-within Difference Difference groups frol Control from CoNtrol P < .05 P < .01 LABCAT CC4.43 Source-Source of Variation Treatments-between groups 27-JUN-2002 418-028:PAGE K-150 Study Report for Clinical Chemistry SUMMARY REPORT PERIOD : TERMINAL STUDY lO; ARGUS 418-028 STUDY NO: 060-069 ANALYSIS OF VARIANCE FOLLOVEOBY OUNNETT'S PROCEOURE SEX: FENALE Albumin/Globulin Group N Total Mean Ratio Std. DUMMETT'S Oev. 't ' DUNNETT'S RANGES LO -95%- HZ tO -99%- HI Source Degree fdm 1-F 10 21.7 2.2 0.28 TREATMENTS 4 2-F 10 26.1 2.6 0.43 3.07 1.8 2.5* 1.7 2.6 ERROR 45 3-F 10 22.3 2.2 0.E1 0.42 1.8 2.5 1.7 2.6 4-F 10 23.7 2.4 0.26 1.39 1.8 2.5 1.7 2.6 TOTAL 49 5-F 10 23.0 2.3 0.37 0.91 1.8 2.5 1.7 2.6 F Ratio = 2.83 'F' table values F.01 = 3.78 F.05 = 2.58 Coeff. Vat. % = 13.7_ Dunnett's 'T' table values P.01 = 3.12 P.05 = 2.51 Sum of Squares 1.16 4.63 5.80 Mean Square 0.E9 0.10 *-Significant Error-within Difference groups from Control P < .05 LABCAT C4.43 Source-Source of Variation Treatmmts-between groups 27- JUN-2002 APPENDIX L STATEMENT OF THE STUDY DIRECTOR 418-028:PAGE L-1 9os or, a Ho_aaraP,A 19044 rehon , 4 -8710 rdef_. (zts) 443.aS87 ARGUS RESEARCH CharlesPdveLr aboratories Discovery and Develt_ment Services PROTOCOL 418-028: ORAL (GAVAGE) COMBINED REPEATED DOSE TOXICITY STUDY OF T-7706 WITH THE REPRODUCTION/ DEVELOPMENTAL TOXICITY SCREENING TEST SPONSOR'S STUDY NUMBER: T-7706.1 STATEMENT OF THE STUDY DIRECTOR This final report accurately reflects the raw data obtained during the performance of the study. No deviations from the U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations; Final Rule a, the Japanese Ministry of Health and Welfare (MHW) Good Laboratory Practice Standard for Safety Studies on Drugs b, the Organisation for Economic Co-operation and Development (OECD). The Revised OECD Principles of Good Laboratory Practices c and the Organisation for Economic Co-operation and Development (OECD), The OECD Guideline for Testing of Chemicals a occurred that affected the quality or integrity of the study. Ra_i_Snd G. York, Phi, _ABT Date Associate Director of R_"g-earch Study Director Argus Research a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance No. 21, March 26, 1997. c. Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97) 186/Final]. d. Organisation for Economic Co-operation and Development (1996). OECD Guideline for Testing of Chemicals. Section 4, No. 422: Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test, adopted 22 March 1996. APPENDIX M QUALITY ASSURANCE STATEMENT 418-028:PAGE M- 1 9oss_.,_ _.,, _agA. PA19044 r_ _2_s)4_ 7_0 r_fax: C_ts_443-8s87 ARGUS RESEARCH CharlesRiverLaboratories DiscoverayndDevelopmenSt ervices QUALITY ASSURANCE STATEMENT Argus Protocol: 418-028 Sponsor's Study Number: T-7706.1 Study Director: Raymond G. York, Ph.D., DABT The protocol, critical phases, raw data and draft final report were inspected by the Quality Assurance Unit (QAU), to assure conformance with: Organisation for Economic Co-operation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97) 186/Final]. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. The undersigned indicate that the report is an accurate representation of the raw data. Data provided by the Sponsor or a subcontractor were not audited by the Argus Research Quality Assurance Unit. 418-028:PAGE M-2 The QAU inspection and report audit dates are listed below: Inspection Phase Protocol Test Substance Inspection Date(s) 22 MAR 02 Date(s) Findings Submitted to Study Director 22 MAR 02 Administration Test Substance 04 APR 02 05 APR 02 Preparation Motor Activity Blood Collection Fetal Blood Collection Fetal Liver Collection Satellite Caesarean- 05 APR 02 08 MAY 02 09 MAY 02 09 MAY 02 09 MAY 02 06 APR 02 16 MAY 02 10 MAY 02 13 MAY 02 13 MAY 02 Sectioning Male Sacrifice Male Blood Collection 09 MAY 02 14 MAY 02 14 MAY 02 13 MAY 02 14 MAY 02 14 MAY 02 Sperm Evaluation Litter Observations Functional Observational 14 MAY 02 17 MAY 02 14 MAY 02 17 MAY 02 Battery Blood Collection Dam/Litter Sacrifice In-Life Data 24 MAY 02 30 MAY 02 30 MAY 02 11-20 SEP 02 24 MAY 02 04 JUN 02 04 JUN 02 23 SEP 02 Necropsy Data Formulation Data 13-20 SEP 02 17 SEP 02 20 SEP 02 17 SEP 02 Report Tables 16-23 SEP 02 24 SEP 02 23 SEP 02 24 SEP 02 Report Text 19,22-23 SEP 02 25 SEP 02 23 SEP 02 25 SEP 02 Revised Report 06 NOV 02 06 NOV 02 Date(s) Findings Submitted to Management 22 MAR 02 05 APR 02 06 APR 02 16 MAY 02 10 MAY 02 13 MAY 02 13 MAY 02 13 MAY 02 14 MAY 02 14 MAY 02 14 MAY 02 17 MAY 02 24 MAY 02 04 JUN 02 04 JUN 02 23 SEP 02 20 SEP 02 17 SEP 02 23 SEP 02 24 SEP 02 23 SEP 02 25 SEP 02 06 NOV 02 Matthew J. Vaneman, B.S. Date Manager of Regulatory Compliance