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__- ~~ 10) OECD 41400PPTS 870.3700, Prenatal development toxicity (teratology), 4 18-023 SANITIZED Study Title Oral (Gavage) Developmental Toxicity Study of Potassium Perfluorobutane Sulfonate (PFBS) in Rats Sponsor's Study Number: T-7485.12 Data Requirement U.S. Environmental Protection Agency Pesticide Assessment Guidelines Subdivision F, 83-3 U.S. Environmental Protection Agency Toxic Substances Control Act Test Guidelines Health Effects Testing Guidelines - 798.4900 Author Raymond G. York, Ph.D., DABT (Study Director) Studv Completed On 18 April, 2002 (Final Report) Performing: Laboratorv Argus Research 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Laboratory Project ID Argus Research, Protocol Number: 4 18-023 Page 1 of 162 ARGUS 418-023 Confidentiality Page This is a blank page to be inserted by the Sponsor. Page 2 ARGUS 418-023 GOOD LABORATORY PRACTICE STATEMENT This study was conducted according to U.S. Environmental Protection Agency (EPA FIFRA/TSCA) "Good Laboratory Practice Standards; Final Rule" (40CFR Part 160/792), the Organisation for Economic Co-operation and Development (OECD) and the Japanese Ministry of Agriculture, Forestry and Fisheries (MAFF) "Good Laboratory Practice Standards" (59 NohSan No. 3850). Any areas of noncompliance are documented in the study record. No deviations existed that affected the validity of the study. Argtk-kesearch L J Paul H. Lieder, Ph.D., DABT Date 3M Corporate Toxicology Page 3 ARGUS 418-023 Flagging Page This is a blank page to be supplied by the Sponsor Page 4 ARGUS 418-023 TITLE: ORAL (GAVAGE) DEVELOPMENTALTOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS SPONSOR'S STUDY NUMBER: T-7485.12 TABLE OF CONTENTS SUBJECT I. SUMMARY AND CONCLUSION A. Methods B. Results C. Conclusion II. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study B. Test Substance Information C. Vehicle Information D. Test Substance Preparation and Storage Conditions E. Test System F. Husbandry G. Methods m. RESULTS A. Mortality, Premature Deliveries and Clinical and Necropsy Observations B. Maternal Body Weights, Body Weight Changes and Gravid Uterine Weights C. Maternal Absolute (g/day) and Relative (g/kg/day) Feed. Consumption Values PAGE 8 8 9 9 10 10 12 12 13 14 16 17 23 23 24 24 Page 5 ARGUS 4 18-023 SUBJECT PAGE D. Caesarean-Sectioningand Litter Observations 25 E. Fetal Alterations 25 REFERENCES 29 APPENDIX A - REPORT FIGURE 31 Figure 1. Maternal Body Weights 32 APPENDIX B - REPORT TABLES 33 Table 1. Clinical Observations - Summary 34 Table 2. Necropsy Observations - Summary 35 Table 3. Maternal Body Weights - Summary 36 Table 4. Maternal Body Weight Changes - Summary 38 Table 5. Maternal Absolute Feed Consumption Values (g/day) - Summary 39 Table 6. Maternal Relative Feed Consumption Values (g/kg/day) - Summary 40 Table 7. Caesarean-SectioningObservations - Summary 41 Table 8. Litter Observations (Caesarean-DeliveredFetuses) - Summary 42 Table 9. Fetal Alterations - Summary 43 Table 10. Fetal Gross External Alterations - Summary 44 Table 11. Fetal Soft Tissue Alterations - Summary 45 Table 12. Fetal Skeletal Alterations - Summary 46 Table 13. Fetal Ossification Sites - Caesarean-DeliveredLive Fetuses (Day 21 of Gestation) - Summary 48 Table 14. Clinical Observations - Individual Data 49 Table 15. Necropsy Observations - Individual Data 53 Table 16. Maternal Body Weights - Individual Data 57 Page 6 SUBJECT ARGUS 418-023 PAGE Table 17. Maternal Feed Consumption Values - Individual Data 65 Table 18. Caesarean-SectioningObservations - Individual Data 69 Table 19. Litter Observations (Caesarean-DeliveredFetuses) - Individual Data 73 Table 20. Fetal Sex, Vital Status and Body Weight - Individual Data 77 Table 21. Fetal Alterations - Individual Data 85 APPENDIX C - PROTOCOL 94 APPENDIX D - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILTY 125 APPENDIX E - CERTIFICATE OF ANALYSIS 127 APPENDIX F - ANALYTICAL REPORT 130 APPENDIX G - ENVIRONMENTAL AND HUSBANDRY REPORTS 141 APPENDIX H - QUALITY ASSURANCE STATEMENT 160 Page 7 ARGUS 418-023 TITLE: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PEEUOROBUTANE SULFONATE (PFBS) IN RATS ARGUS RESEARCH PROTOCOL NUMBER: 418-023 SPONSORS STUDY NUMBER: T-7485.12 I. SUMMARY AND CONCLUSION A. Methodsa One hundred Crl:CD@(SD)IGSBR VAF/Plus@presumed pregnant female rats were randomly assigned to four dosage groups (Groups I through IV),25 rats per dosage group. The test substance, Potassium Perfluorobutane Sulfonate (PFBS or T-7485), or the vehicle, aqueous 0.1% carboxymethylcellulose, were administered via gavage once daily to these female rats on days 6 through 20 of gestation (DGs 6 through 20). Dosages of 0 (Vehicle), 100,300 and 1000were administered at a dosage volume of 10 mL/kg, adjusted daily on the basis of the individual body weights recorded before intubation and administered at approximately the same time each day. The female rats were observed for viability at least twice each day of the study. Rats were also examined for clinical observations of effects of the test substance, abortions, premature deliveries and deaths before and approximately 60 +_ 10 minutes after dosage administration and on the day of scheduled sacrifice. Body weights were recorded daily during the dosage period and prior to sacrifice. Feed consumption values were recorded on DGs 0, 6,9, 12, 15, 18, 20 and 21. All surviving rats were sacrificed on DG 21, Caesarean-sectioned and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The gravid uterus was excised and weighed. The number of corpora lutea in each ovary was recorded. The uterus of each rat was excised and examined for pregnancy, number and distribution of implantations, live and dead fetuses and early and late resorptions. Each fetus was identified, weighed and examined for sex and gross external alterations. Approximately one-half of the fetuses in each litter were examined for soft tissue alterations and the remaining fetuses were examined for skeletal alterations. a. Detailed descriptions of all procedures used in the conduct of this study are provided in the appropriate sections of this report and in APPENDIX C (PROTOCOL). Page 8 ARGUS 4 18-023 B. Results Two rats in the 1000 mg/kg/day dosage group were found dead. The death of one of these rats was attributed to an intubation accident. No cause of death was determined for the other rat that was found dead in this dosage group, but the death was considered unrelated to the test substance because it was a single event. One rat in each of the 0 (Vehicle) and 300 mg/kg/day dosage groups began delivery before Caesareansectioning on day 21 of gestation (DG 21) and were sacrificed. All other rats survived until scheduled sacrifice. All clinical and necropsy observations were considered unrelated to the test substance. Maternal body weight gains were significantly reduced on DGs 6 to 9 and 18 to 21 in the 1000 mgkg/day dosage group. As a result of these reductions, maternal body weight gains were significantly reduced for the entire gestation period (DGs 0 to 21) and maternal body weights were significantly reduced on DGs 20 and 21 in this dosage group. Gravid uterine weights were slightly reduced (91% of control value) in the 1000 mg/kg/day dosage group, but this reduction was not statistically significant. Absolute (g/day) and relative (g/kg/day) feed consumption values were significantly reduced on DGs 9 to 12 (relative only) and 18 to 21 in the 1000mgkglday dosage group. As a result of these reductions, absolute and relative feed consumption values were significantly reduced in the 1000 mg/kg/day dosage group for the entire dosage period (calculated as DGs 6 to 21). Fetal body weights (total, male and female) were significantly reduced in the 1000 mg/kg/day dosage group, compared to the control group value. No other Caesarean-sectioning or litter parameters were affected by dosages of the test substance as high as 1000 mgkg/day. No gross external, soft tissue or skeletal fetal alterations (malformations or variations) were considered test substance related. C. Conclusion On the basis of these data, the maternal no-observable-adverse-effect-level(NOAEL) of PFBS is 300 mg/kg/day (the 1000 mg/kg/day dosage caused reductions in body weight gain and reduced absolute and relative feed consumption values). The developmental NOAEL is also 300 mg/kg/day (the 1000 mg/kg/day dosage caused reductions in fetal body weights). Alan M. Hoberman, Ph.D., DABT Date Director of Research Rvond G. Y&kdh.D., DABT Date Ass ciate Director of Research and Study Director Page 9 ARGUS 4 18-023 11. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study A.l. Sponsor 3M Corporate Toxicology, 3M Center, Building 220-2E-02, St. Paul, Minnesota 55 144-1000 A.2. Testing Facility Argus Research, 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 A.3. Study Number 418-023 A.4. Sponsor's Study Number T-7485.12 A S . Purpose of the Study The purpose of this study was to evaluate the developmental toxicity (embryo-fetal toxicity and teratogenic potential) of Potassium Peffluorobutane Sulfonate (PFBS) administered orally via gavage to Crl:CD@(SD)IGSBR VAF/Plus@presumed pregnant female rats. A.6. Study Design The requirements of the U.S. Environmental Protection Agency (EPA)''*2', the Organization for Economic Cooperation and Development (OECD)'3', and the Japanese Ministry of Agriculture, Forestry and Fisheries (MAFF)'4' were used as the basis for study design. A.7. Regulatory Compliance The study was conducted in compliance with Good Laboratory Practice (GLP) regulations of the U.S. Environmental Protection A ency (EPA)'5*6't,he Organization for Economic Cooperation and Development (OECD)", and the Japanese Ministry of Agriculture, Forestry and Fisheries (MAFF)'*'. 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. Page 10 ARGUS 418-023 A.8. Ownership of the Study The Sponsor owns the study. All raw data, analyses, reports and preserved tissues are the property of the Sponsor. A.9. Study Monitor Paul H. Lieder, Ph.D., DABT A.lO. Study Director Raymond G. York, Ph.D., DABT (Associate Director of Research, Argus Research, 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297) A.ll. Technical Performance John F. Barnett, B.S. (Director of Laboratory Operations) Todd J. Killino, B.S. (Research Associate) Timothy J. Brommer, B.S. (Laboratory Technician) Brian K. Kelsch (Necropsy Laboratory Technician) Christopher K. Ruppert, B.S. (FormulationLaboratory Technician) A.12. Report Preparation Raymond G. York, Ph.D., DABT Jo Ann Frazee, M.S. (Study Coordinator) Cindy L. Mininger, B.S. (Data Management Specialist) Natalie A. Farst (Report Administrator) Georgia Y. Burnett, A.A.S. (Report Administrator) A.13. Report Review Valerie A. Sharper, M.S. (Director of Study Management) A.14. Date Protocol Signed 16 February 200 1 A.15. Dates of Technical Performance Rat Arrival Cohabitation Period DG Oa Dosage Period (DGs 6 through 20) Caesarean-Sectioning Period (DG 2 1) 13 FEB 01 18 FEB 01 PM - 23 FEB 01 AM 19 FEB 01 - 23 FEB 01 25 FEB 01 - 14 MAR01 12 MAR 01 - 15 MAR 01 a. DG is an abbreviation used for day of (presumed) gestation. Page 11 ARGUS 4 18-023 A.16. Records Maintained The original report, raw data and reserve samples of the test substance and vehicle 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. Unused bulk test substance will be returned to the Sponsor. B. Test Substance Information B.1. Description Potassium Perfluorobutane Sulfonate (PFBS or T-7485)- a white powder B.2. Lot Number 5 B.3. Date Received and Storage Conditions The test substance was received on 23 February 2001, and stored at room temperature. B.4. 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 preparation and dosage. TyvekB sleeves and a half-face respirator were worn and procedures were conducted in a chemical fume hood when using the bulk test substance. B.5. Analysis of Activity Information regarding the identity, composition, method of synthesis, strength and purity of the test substance is on file with the Sponsor. A Certificate of Analysis is available in APPENDIX E. C. Vehicle Information C.l. Description Aqueous 0.1% carboxymethylcellulose"(CMC, medium viscosity) prepared using carboxymethylcellulose, an off-white powder, and reverse osmosis membrane processed deionized water (R.O. deionized water) a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PRTOCEDURES OF THE TESTING FACILTY), item 1. Page 12 ARGUS 418-023 C.2. Lot Number 49H 1332 C.3. Date Received and Storage Conditions The powdered carboxymethylcellulosewas received from Sigma Chemical Co., St. Louis, Missouri, on 19 October 1999, 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. C.4. 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 when handling the vehicle. C.5. Analysis of Purity Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the vehicle that would have interfered with the results of this study. D. Test Substance Preparation and Storage Conditions Suspensions of the test substance were prepared approximately every 10 days at the Testing Facility. Prepared test substance and vehicle formulations were stored at 2C to 8C. Page 13 D.l. Sample Information ARGUS 418-023 R.O. Deionized Water 1 g 20 MAR 01 a. James D. Johnson, Southern Research Institute, 2000 Ninth Avenue South, Birmingham, Alabama 35205. b. Duplicate samples were taken from each concentration of the first and last preparation on the day prepared. One sample of each set was shipped for analysis. The remaining samples were retained at 2C to 8C' at the Testing Facility as backup samples. D.2. Analytical Results The 10 and 30 mg/mL samples were all found to be within -e 10% of the target concentrations; the 100mg/mL samples were 86.3% (first preparation) and 106% (last preparation) of target. Information to document the homogeneity .and concentration of the test substance in the prepared vehicle over the range of concentrations used in this study and stability data for prepared formulations bracketing the range of concentrations in this study, are available in APPENDIX F. Result of the bulk test substance analysis is available in APPENDIX F. E. Test System E.l. Species Rat E.2. Strain Crl:CD@(SD)IGSBR VAF/Plus@ E.3. Supplier (Source) Charles River Laboratories, Inc., Raleigh, North Carolina E.4. Sex Female (Note: Male rats were used only for the purposes of breeding and are not considered part of the Test System.) Page 14 ARGUS 418-023 E.5. Rationale for Test System The Crl:CD@(SD)IGSBR VAFPlusB rat was selected as the Test System because: 1) it is one mammalian species accepted and widely used throughout industry for nonclinical studies of developmental toxicity (embryo-fetal toxicity/teratogenicity); 2) this strain has ebxepenerdieenmcoenesxtirsatteadt tthoebTeessetninsgitiFvaectiolitdye(9v-e11l)o.pmentaltoxins; and 3) historical data and E.6. Test System Data Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) on the Day after Arrival Weight (g) at Study Assignment 140 11 DEC 00 65 days 187 - 227 220 - 244 E.7. Breeder Male Rat Data Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) on the Day after Arrival Weight (g) at Cohabitation 250 17 JUL 00 79 days 310 - 388 526 - 876 E.8. Method of Randomization Upon arrival, male and female rats were assigned to individual housing on the basis of computer-generated random units. Healthy, mated female rats were assigned to four dosage groups (Groups I through IV), 25 rats per dosage group, using a computer- generated (weight-ordered) randomization procedure based on body weights recorded on DG 0. E.9. System of Identification Rats were permanently identified with a Monel@self-piercingear tag (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats were given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats were assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to study. Cage tags were marked with the study number, permanent rat number, sex, test substance identification, generation and dosage level. Page 15 ARGUS 4 18-023 F. Husbandry F.l. Research Facility Registration USDA Registration No. 14-R-0144under the Animal Welfare Act, 7 U.S.C. 213 1 et seq. F.2. Study Rooms The study rooms were maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum of ten changes per hour of 100%fresh air that had been passed through 99.97% HEPA filters. Room temperature and humidity were monitored constantly throughout the study. Room temperature was targeted at 64F to 79F (18C to 26C);relative humidity was targeted at 30% to 70%=. F.3. Housing Rats were individually housed in stainless steel, wire-bottomed cages except during the cohabitation period. During cohabitation, each pair of male and female rats was housed in the male rat's cage. All cage sizes and housing conditions were in compliance with the Guidefor the Care and Use of Laboratory F.4. Lighting An automatically-controlled fluorescent light cycle was maintained at 12-hours light: 12-hours dark, with each dark period beginning at 1900hours EST. F.5. Sanitization Cage pan liners were changed approximately four times weekly. Cages were changed approximately every other week. F.6. Feed Rats were given ad libitum access to Certified Rodent Diet@#5002 (PMI Nutrition International, St. Louis, Missouri) in individual feeders. F.7. Feed Analysis Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration limits for certified feed or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data and APPENDIX G. a. See APPENDIX G (ENVIRONMENTALAND HUSBANDRY REPORTS). Page 16 ARGUS 418-023 Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the feed that would have interfered with the results of this study. F.8. Water Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad libitum from an automatic watering access system. Chlorine was added to the processed water as a bacteriostat. F.9. Water Analysis The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, Pennsylvania) and monthly for possible bacterial contamination (Analytical Laboratories, Inc., Chalfont, Pennsylvania). Copies of the results of the water: analyses are available in the raw data and APPENDIX G. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the water that would have interfered with the results of this study. G. Methods G.l. Dosage Administration 19151 - 19175 Iv 1000 100 10 25 19176- 19200 a. The test substance was considered 100% pure for the purpose of dosage calculations. G.2. Rationale for Dosage Selection Dosages were selected on the basis of a dosage-range study (Argus Research, Protocol 418-023P). It was anticipated that the highest dosage level selected would induce some overt developmental and/or maternal mortality, the intermediate dosage levels would produce minimal observed toxic effects and the lowest dosage level would not produce any evidence of either maternal or developmental toxicity. In the 418-023P study, all rats survived to scheduled sacrifice. Clinical observations considered test substance-related were limited to urine-stained abdominal fur in four rats in the 2000 mg/kg/day dosage group, Rats in the 2000 mg/kg/day dosage group lost Page 17 ARGUS 4 18-023 weight on DGs 6 to 9 and body weight gains were reduced at several intervals tabulated during the dosage period. Reflecting these effects of the test substance, body weight gain was reduced in the 2000 mg/kg/day dosage group for the entire treatment (DGs 6 to 21) and gestation (DGs 0 to 21) periods. Absolute and relative feed consumption values were reduced in the 2000 mg/kg/day dosage group on DGs 6 to 9,9 to 12,12 to 15, 15 to 18 and 18 to 2 1, and for the entire treatment and gestation periods. Fetal body weights were reduced in the 2000 mg/kg/day dosage group, as compared with the control group. No other Caesarean-sectioning or litter parameters were affected by dosages of the test substance as high as 2000 mg/kg/day. No fetal gross alterations occurred. G.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 possible route of human exposure. 6.4. Frequency of Administration Appropriate dosages of the test substance or vehicle were administered orally (via gavage) once daily to rats on DGs 6 through 20. 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. G.5. Method of Study Performance After acclimation, 140 healthy virgin female rats were placed into cohabitation with 140 breeder male rats, one male rat per female rat. The cohabitation period consisted of a maximum of five days. Mating performance was evaluated daily during the cohabitation period. Female rats with spermatozoa observed in a smear of the vaginal contents andor a copulatory plug in situ were considered to be at DG 0 and assigned to individual housing. Rats were observed for viability at least twice each day of the study. Rats were also examined for clinical observations and general appearance weekly during the acclimation period and on DG 0. Observations for clinical signs of effects of the test substance, abortions, premature deliveries and deaths were also made daily before and approximately 60 f 10 minutes after dosage administration and on the day of scheduled sacrifice. Body weights were recorded weekly during the acclimation period, on DG 0, daily during the dosage period and prior to sacrifice. Feed consumption values were recorded on DGs 0, 6, 9, 12, 15, 18, 20 and 21. Page 18 ARGUS 418-023 6.6. Gross Necropsy In order to minimize bias, Caesarean-sectioning and subsequent fetal observations were conducted without knowledge of dosage group. All surviving rats were sacrificed by carbon dioxide asphyxiation on DG 21, Caesarean-sectioned and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The gravid uterus was excised and weigheda. Uteri of apparently nonpregnant rats were examined while being pressed between glass plates to confirm the absence of implantation sites. The number of corpora lutea in each ovary was recorded. The uterus of each rat was excised and examined for pregnancy, number and distribution of implantations, live and dead fetuses and early and late resorptions. An early resorption was defined as one in which organogenesis was not grossly evident. A late resorption was defined as one in which the occurrence of organogenesis was grossly evident. A live fetus was defined as a term fetus that responded to stimuli. Nonresponding term fetuses were considered to be dead (there were no dead fetuses). Dead fetuses and late resorptions were differentiated by the degree of autolysis present; marked to extreme autolysis indicated that the fetus was a late resorption. Each fetus was removed from the uterus, placed in an individual container and identified with a tag noting the study number, litter number, uterine distribution and fixative. Each fetus was subsequently weighed and examined for sex and gross external alterations. Live fetuses were sacrificed by an intraperitoneal injection of sodium pentobarbital. Late resorptions were examined to the extent possible. Approximately one-half of the fetuses in each litter were examined for soft tissue alterations using a variation of the microdissection technique of staple^"^). These fetuses were fixed in Bouin's solution and the heads were subsequently examined by free-hand sectioning; head sections were retained in alcohol. The decapitated carcasses were discarded. The remaining fetuses (approximately one-half of the fetuses in each litter) were examined for skeletal alterations (bone and cartilage) after staining with alizarin red $I4). The fetuses were initially fixed in alcohol; skeletal preparations were retained in glycerin with thymol added as a preservative. Rats that died or were sacrificed because of premature delivery were examined for the cause of death on the day the observation was made. The rats were examined for gross lesions. Pregnancy status and uterine contents of female rats were recorded. Gravid uterine weights were recorded unless precluded by autolysis. Delivered pups and fetuses in uteri were examined to the extent possible, using the methods described for term fetuses. Uteri of apparently nonpregnant rats will be examined while being pressed between glass plates to confirm the absence of implantation sites. a. See APPENDIX D, item 2. Page 19 ARGUS 418-023 6.7. Data Collection and Statistical Analyses Data generated during the course of this study were recorded either by hand or using the Primedica Argus Automated Data Collection and Management System and the Vivarium Temperature and Relative Humidity Monitoring System. All data were tabulated, summarized and/or statistically analyzed using the Primedica Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microsofi Excel [part of Microsoft Office 97 (version SR-2)] andor the SAS System (version 6.12). Page 20 ARGUS 418-023 Averages and percentages were calculated. Litter values were used where appropriate. The following schematic represents the statistical analyses of data: Tvpe of Testa I. Parametric II. Nonparametricb A. Bartlett's Test' A. Kruskal-WallisTest (175% ties) I Significant at p10.05 Dunnett's Test Not Significant B. Fisher's Exact Test (>75% ties) III. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution ___--___________----- a. Statistically significant probabilities are reported as either p10.05 or p10.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. Page 21 ARGUS 418-023 Clinical observation and other proportion data were analyzed using the Variance Test for Homogeneity of the Binomial Distrib~tion('~). Continuous data (e-g.,maternal body weights, body weight changes, feed consumption values and litter averages for percent male fetuses, percent resorbed conceptuses, fetal body weights, fetal anomaly data and fetal ossification site data) were analyzed using Bartlett's Test of Homogeneity of Variances(16)and the Analysis of Variance'"), when appropriate [i.e., Bartlett's Test was not significant (p>O.OOl)]. If the Analysis of Variance was significant (pl0.05),Dunnett's Test(18)was used to identify the statistical significance of the individual groups. If the Analysis of Variance was not appropriate [i.e., Bartlett's Test was significant Cp<O.OOl)], the Kruskal-Wallis Test'") was used, when less than or equal to 75% ties were present. In cases where the Kruskal-Wallis Test was statistically significant (p<O.OS), Dunn's Method of Multiple Comparisons'20'was used to identify the statistical significance of the individual groups. If there were greater than 75% ties, Fisher's Exact Test(2')was used to analyze the data. Count data obtained at Caesarean-sectioningof the dams were evaluated using the procedures described above for the Kruskal-Wallis Test("). Page 22 ARGUS 418-023 111. RESULTS - - A. Mortality, Premature Deliveries and Clinical and Necropsy Observations (Summaries Tables 1 and 2; Individual Data Tables 14 and 15) A.l. Mortality Two rats in the 1000 mg/kg/day dosage group were found dead. The death of one of these rats was attributed to an intubation accident. No cause of death was determined for the other rat that was found dead in this dosage group, but the death was considered unrelated to the test substance because it was a single event. One rat in each of the 0 (Vehicle) and 300 mg/kg/day dosage groups began delivery before Caesareansectioning on day 21 of gestation (DG 21) and were sacrificed. Observations in these rats are described below. All other rats survived until scheduled sacrifice. Rat 19181 in the 1000 mg/kg/day dosage group was found dead on day 18 of presumed gestation (DG 18) before the 13th daily dosage. This rat was cold to touch and dehydrated on DG 17. This rat had a variable pattern of weight gains and losses after DG 6 and weighed approximatelythe same amount on DG 16 as on DG 6. There was a weight loss from DG 16 to 17. Feed consumption values were comparable to other nonpregnant rats in this dosage group. All tissues examined appeared normal at necropsy. The rat was not pregnant. No cause of death could be determined. Rat 19198 in the 1000 mg/kg/day dosage group was found dead on DG 17, seven minutes after the 12th daily dosage. Clinical signs observed prior to being found dead were all normal. After an initial weight loss on DGs 6 to 7, this rat gained weight throughout the study. Feed consumption values were comparable to other rats in the dosage group. Necropsy revealed a 0.1 cm diameter perforation of the esophagus and 3 mL of red substance in the thoracic cavity. All other tissues examined at necropsy appeared normal. The litter consisted of 15 fetuses and one early resorption in utero. All fetuses appeared normal at gross external examination; further examination was precluded by early developmental age. Based on the necropsy results, this death was attributed to an intubation error. Rat 19111 in the vehicle control group delivered two pups on DG 21 and was euthanized. Clinical signs observed prior to delivery were all normal. Body weight gains and feed consumption values were comparable to other rats in the dosage group. All tissues examined at necropsy appeared normal. There were 13 fetuses and one early resorption in utero. All pups and fetuses appeared normal at gross external, soft tissue and skeletal examinations. Rat I9 157 in the 300 mg/kg/day dosage group delivered one pup on DG 21 and was euthanized. Clinical signs observed prior to delivery were all normal. Body weight gains and feed consumption values were comparable to other rats in the dosage group. All tissues examined at necropsy appeared normal. There were 16 fetuses in utero. All Page 23 ARGUS 418-023 fetuses and the pup appeared normal at gross external, soft tissue and skeletal examinations. A.2. Clinical and Necropsy Observations All clinical observations were considered unrelated to the test substance because: 1) the incidences were not dosage-dependent; or 2) the observations occurred in only one or three rats. These clinical observations included localized alopecia on the limbs in rats in the 0 (Vehicle), 100 and 300 mg/kg/day dosage groups and cold to touch and dehydration in the 1000 mg/kg/day dosage group rat that died as the result of an intubation error. The only adverse necropsy observations were perforation of the esophagus and red fluid in the thoracic cavity in the 1000 mg/kg/day dosage group rat that died as the result of an intubation error. - - B. Maternal Body Weights, Body Weight Changes and Gravid Uterine Weights (Figure 1; Summaries Tables 3 and 4 Individual Data Table 16) Maternal body weight gains were significantly reduced (p#O.OS or p#O.Ol) on DGs 6 to 9 and 18 to 21 in the 1000 mg/kg/day dosage group. As a result of these reductions, maternal body weight gains were significantly reduced (p#O.Ol) in the 1000 mg/kg/day dosage group for the entire gestation period (DGs 0 to 21). When the DG 21 body weight was corrected for gravid uterine weight (DG 21C), maternal body weight gains for the entire dosage period (calculated as DGs 6 to 21C) were significantlyreduced (p10.01). Maternal body weights were significantly reduced (p#0.05 or p#O.Ol) in the 1000 mg/kg/day dosage group on DGs 20 and 21. Gravid uterine weights were slightly reduced (9 1% of control value) in the 1000 mg/kg/day dosage group, but this reduction was not statistically significant. Body weights, body weight gains and gravid uterine weights were unaffected by dosages of the test substance as high as 300 mg/kg/day. Maternal body weight gains in the 100 and 300 mg/kg/day dosage groups were significantlyreduced @#0.05 or ~ 1 0 . 0 1o)n DGs 18 to 2 1 and this resulted in a significant reduction on DGs 6 to 2 1. These reductions were not considered test substance-relatedbecause they were single occurrences and were not significant ( p 0 . 0 5 )when the body weights were corrected for uterine contents (calculated DGs 6 to 2 1). - - C. Maternal Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values (Summaries Tables 5 and 6; Individual Data Table 17) Absolute (g/day) and relative (g/kg/day) feed consumption values were significantly reduced @#O.Ol) on DGs 18 to 21, and the relative feed consumption value was significantly reduced (p#0.05)on DGs 9 to 12 in the 1000 mg/kg/day dosage group. ASa result of these reductions, absolute and relative feed consumption values were Page 24 ARGUS 418-023 significantlyreduced (p#O.Ol)in the 1000 mg/kg/day dosage group for the entire dosage period (calculated as DGs 6 to 21). Absolute and relative feed consumption values were unaffected by dosages of the test substance as high as 300 mg/kg/day. Absolute andor relative feed consumption values were significantly reduced (p#0.05) on DGs 18 to 21 in the 100 and 300 mg/kg/day dosage groups but the reductions were not considered test substance-relatedbecause they were single occurrences and did not persist. - - D. Caesarean-Sectioningand Litter Observations (Summaries Tables 7 and 8; Individual Data Tables 18 through 20) Caesarean-sectioningobservations were based on 24,22,23 and 2 1 pregnant rats that survived to DG 21 in the 0 (Vehicle), 100,300 and 1000mg/kg/day dosage groups, respectively. Fetal body weights (total, male and female) were significantlyreduced @#O.Ol)in the 1000 mg/kg/day dosage group, compared to the control group value. No other Caesarean-sectioningor litter parameters were affected by dosages of the test substance as high as 1000 mg/kg/day. The litter averages for implantations, litter sizes, live fetuses, early and late resorptions, percent resorbed conceptuses, and percent live male fetuses were comparable among the four dosage groups and did not significantly differ. No dam had a litter consisting of only resorbed conceptuses, there were no dead fetuses and all placentae appeared normal. The litter average for corpora lutea was significantly reduced @#O.Ol)in the 300 mg/kg/day dosage group but the reduction was not considered treatment-related because: 1) it was not dosage dependent; and 2) corpora lutea formation occurred prior to dosage administration. E. - Fetal Alterations (Summaries Tables 9 through 13; Individual Data - Table 21) Fetal alternations were defined as: 1) malformations (irreversible changes that occur at low incidences in this species and strain); or 2) variations (common findings in this species and strain and reversible delays or accelerations in development). Litter averages were calculated for specific fetal ossification sites as part of the evaluation of the degree of fetal ossification. Fetal evaluations were based on 358, 316,331 and 311 live, DG 21 Caesarean-delivered fetuses in 24, 22,23 and 21 litters in the 0 (Vehicle), 100,300 and 1000 mg/kg/day dosage groups, respectively. Each of these fetuses was examined for gross external alterations. Of these respective fetuses, 173, 150, 157 and 150 fetuses were examined for soft tissue alterations, and 185, 166, 174 and 161 fetuses were examined for skeletal . alterations and fetal ossification site averages. Page 25 ARGUS 418-023 E.l. - Summary of (Summary Fetal Alterations Table 9; Individual Data - Table 21) The number of litters with fetuses with alterations numbered 4 (16.7%), 8 (36.4%), 3 (13.0%) and 3 (14.3%), in the 0 (Vehicle), 100,300 and 1000mg/kg/day dosage groups, respectively. The numbers of fetuses with any alteration observed were 4 (1.1%), 13 (4.1%)**,3 (0.9%)and 3 (1.0%),and the percentages of fetuses with any alteration per litter were 1.2,4.3,0.8 and 1.0 in these same respective dosage groups. No gross external, soft tissue or skeletal fetal alterations (malformations or variations) were caused by dosages of the test substance as high as 1000 mg/kg/day. There were no dosage-dependentor significant differences in the litter or fetal incidences of any gross external, soft tissue or skeletal alterations. The significant increase (p#O.Ol) in the percentage of fetuses with any alteration in the 100 mg/kg/day dosage groups was not considered treatment-related because it was not dosage-dependent. All fetal alterations in this study are described in the following information. - - E.2. Fetal Gross External Alterations (Summary Table 10; Individual Data Table 21) No gross external fetal alterations occurred. - - E.3. Fetal Soft Tissue Alterations (Summary Table 11; Individual Data Table 21) E.3.a. Malformations One control group fetus (19118-10) had an absent left eye. No additional alterations occurred in this fetus. E.3.b. Variations E.3.b.l. Eyes Folded retina, a variation usually attributable to processing, occurred in one fetus (19143-19) in the 100 mg/kg/day dosage group. No additional alterations occurred in this fetus. E.3.b.2. Vessels The umbilical artery descended to the left of the urinary bladder in 1 , 2 , 0 and 0 fetuses from 1 , 2 , 0 and 0 litters from the 0 (Vehcle), 100,300 and 1000 mg/kg/day dosage groups, respectively. No additional alterations occurred in these fetuses. ** Significantly different from the vehcle control group value at p#O.Ol Page 26 ARGUS 418-023 Absent innominate artery occurred in one fetus (19185-8) in the 1000 mg/kg/day dosage group. No additional alterations occurred in this fetus. - - E.4. Fetal Skeletal Alterations (Summaries Tables 12 and 13; Individual Data Table 21) E.4.a. Malformations One fetus in the vehicle control group (19120-5) had a left hemivertebra present as the 14th thoracic vertebra (arch and centrum with attached rib), fused arches of the 13th and 14th thoracic vertebrae, unilateral ossification (left) of the centrum of the 14th thoracic vertebra and not ossified centrum of the 13th thoracic vertebra. This fetus also had a bifid centrum of the 11th thoracic vertebra. One 100 mg/kg/day dosage group fetus (19138-9) had proximally fused 4th and 5th right ribs. This fetus also had a bifid centrum of the 4th thoracic vertebra. E.4.b. Variations As described in the following information, all skeletal variations were reversible delays in fetal ossification(22-23),with the exception of cervical rib, a common variation in this strain of rat(24). E.4.b.l. Vertebrae A bifid centrum in a thoracic vertebra occurred in 2 , 4 , 3 and 1 fetuses in 2,4, 3 and 1 litters in the four respective dosage groups. Fetuses 19120-5 and 19138-9 in the 0 (Vehicle) and 100 mg/kg/day dosage groups, respectively, had additional skeletal alterations, as previously described. E.4.b.2. Ribs A cervical rib at the 7th cervical vertebra, a common variation in this strain of rat(24)w, as present in 0, 3,O and 1 fetuses from 0,2,0 and 1 litters in the four respective groups. No additional alterations occurred in these fetuses. E.4.b.3. Sternum Delayed sternal ossification (fused 1st and 2nd and asymmetric 1st through 3rd sternal centra) occurred in one fetus (19140-12) in the 100 mg/kg/day dosage group. No additional alterations occurred in this fetus. Page 27 ARGUS 418-023 E.4.b.4. Pelvis One fetus (19139-7) in the 100 mgkglday dosage group had an incompletely ossified right pubis. No additional alterations occurred in this fetus. E.4.b.5. Fetal Ossification Site Averages Fetal ossification sites in the hindlimbs (metatarsals and phalanges) were significantly reduced (pLO.01) in the 1000 mg/kg/day dosage group. These reductions in the number of ossification sites are considered reversible delays in ossification associated with the maternal toxicity (reduced body weights, body weight gains and feed consumption) observed at this dosage level. There were no statistically significant or biologically important differences among the four dosage groups in the average numbers of ossification sites per fetus for the hyoid, vertebrae (cervical,' thoracic, lumbar, sacral and caudal), ribs, sternum (manubrium, sternal centers and xiphoid), forelimbs (carpals, metacarpals and phalanges) or hindlimbs (tarsals). Page 28 ARGUS 4 18-023 REFERENCES 1. U.S. Environmental Protection Agency (1998). Health Effects Test Guidelines; Prenatal Developmental Toxicity Study. Office of Prevention, Pesticides and Toxic Substances (OPPTS) 870.3700, August, 1998. 2. U.S. Environmental Protection Agency (1997). Toxic Substances Control Act (TSCA) Test Guidelines; Final Rule. Prenatal Developmental Toxicity, 799.9370 (cross-referencedto OPPTS 870.3700). Federal Register, August 15, 1997. 3. Organization for Economic Cooperation and Development (1981). OECD Guidelinesf o r Testing of Chemicals. Section 4, No. 4 14: Teratogenicity, adopted 12 May 1981. 4. Japanese Ministry of Agriculture, Forestry and Fisheries (1985). Guidance on Toxicology Study Data for Application of Agricultural Chemical Registration. 59 NohSan No. 4200. 5 . U.S. Environmental Protection Agency. Toxic Substances Control Act (TSCA); Good Laboratory Practice Standards; Final Rule. 40 CFR Part 792. 6. U.S. Environmental Protection Agency. Federal Insecticide, Fungicide and Rodenticide Act (FIFRA); Good Laboratory Practice Standards; Final Rule. 40 CFR Part 160. 7. Organization for Economic Cooperation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. 8. Japanese Ministry of Agriculture, Forestry and Fisheries (1984). Good Laboratory Practice Standards. 59 NohSan No. 3850. 9. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 10. 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. 11. Lang, P.L. ( 1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Crl:CD@BRRat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research.) Page 29 ARGUS 418-023 12. Institute of Laboratory Animal Resources (1996). Guidefor the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 13. Staples, R.E. (1974). Detection of visceral alterations in mammalian fetuses. Teratology 9(3):A37-38. 14. Modification of method of Staples, R.E. and Schnell, V.L. (1964). Refinement in rapid clearing technique in the KOH-alizarin red S method for fetal bone. Stain Technol. 39:61-63. 15. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 240-24 1. 16. 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. 17. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 18. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. h e r . Stat. Assoc. 50:1096-1129. 19. Sokal, R.R. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 20. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241-252. 21. Siegel, S. (1956). Nonparametric Statisticsfor the Behavioral Sciences, McGraw-Hill, New York, pp. 96-104. 22. Hannah, R.S. and Moore, K.L. (1971). Effects of fasting and insulin on skeletal development in rats. Teratology 4: 135-140. 23. Collins, T.F.X., Welsh, J.J., Black, T.N., Whitby, K.E. and O'Donnell, M.W., Jr. (1987). Potential reversibility of skeletal effects in rats exposed in utero to caffeine. Fd. Chem. Toxic. 25(9):647-662. 24. Khera, K.S. (1981). Common fetal aberrations and their teratologic significance: a review. Fund. and Appl. Toxicol. 1:13-18. Page 30 ARGUS 418-023 APPENDIX A REPORT FIGURE Page 31 PROTOCOL418-0231ORAL (GAVAGE)DEVELOPMENTAL TOXICITY STUDY OF POTASSIUMPERFLUOROBUTANESULFONATE(PFBS) IN RATS(SPONSOR'S STUDYNUMBER: T-7485.12) 450 400 5ccr ?. w h 350 I4 I- I 12 300 MATERNAL BODY WEIGHTS Figure 1 -TI 0 (VEHICLE) MGIKGIDA --1- 100 MGIKGIDAY 300 MGIKGIDAY -A- 1000 MGIKGIDAY 250 d 200 ~ d 6 f Q --,rT5-7'4 ~ ?4 - -lF---?l 1 5 16 117 -x-119210 211 DAY OF GESTATION 1_1 ** p50.01 ARGUS 418-023 APPENDIX B REPORT TABLES Page 33 PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T - 7 4 8 5 . 1 2 ) TABLE 1 (PAGE 1): CLINICAL OBSERVATIONS - SUMMARY FOUND DEAD 0 0 0 2b, c DELIVERED AND SACRIFICED Id 0 le 0 COLD TO TOUCH o/ 0 o/ 0 o/ 0 1/ lb DEHYDRATION o/ 0 o/ 0 o/ 0 1/ lb LOCALIZED ALOPECIA: LIMBS 261 3 -J/ 1 151 1 o/ 0 -----___________________________________------------------~-------------------------------------------------------------- STATISTICAL ANALYSES OF CLINICAL OBSERVATION DATA WERE RESTRICTED TO THE NUMBER OF RATS WITH OBSERVATIONS. MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP ON DAYS 6 THROUGH 21 OF PRESUMED GESTATION. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. w a. Dosage occurred on days 6 through 20 of presumed gestation. P b . Dam 19181 was found dead on day 18 of gestation. c. Dam 19198 was found dead on day 17 of gestation. d. Dam 19111 delivered and was sacrificed on day 21 of gestation. e. Dam 19157 delivered and was sacrificed on day 21 of gestation. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 2 (PAGE 1 ) : NECROPSY OBSERVATIONS - SUMMARY DELIVERED AND SACRIFICED N le 0 If 0 APPEARED NORMAL N 24e 25 24 f 23c ESOPHAGUS : PERFORATION N 0 0 0 Id THORACIC CAVITY: RED FLUID N 0 0 0 Id ________________________________________-------------------------------------------------------------------------------------------- a. Dosage occurred on days 6 through 20 of presumed gestation. b. Refer to the individual clinical observations table (Table 14) for external observations confirmed at necropsy. c. Dam 19181 was found dead on day 18 of gestation. d. Dam 19198 was found dead on day 17 of gestation. e. Dam 19111 delivered and was sacrificed on day 21 of gestation. f. Dam 19157 delivered and was sacrificed on day 21 of gestation. 0 N w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 3 (PAGE 1): MATERNAL BODY WEIGHTS - SUMMARY DAY 0 DAY 6 DAY 7 DAY 8 ? DAY 9 %wm DAY 10 DAY 1 1 DAY 12 DAY 13 DAY 14 MEANfS.D . MEANfS . D. MEANfS.D. MEANfS .D. MEANfS .D. MEANfS .D. MEAN&. D. MEAN+S .D . MEANfS .D . MEANfS .D. 231.7 f 6.6 262.8 f 8.5 266.2 f 8.8 269.8 f 8.9 273.8 f 9.4 278.4 f 10.3 285.2 f 10.3 291.9 + 10.1 296.2 f 12.2 302.2 f 12.2 232.9 f 6.7 263.2 f 9.9 265.5 f 9.4 269.0 f 10.2 273.3 f 9.9 277.4 f 10.0 283.5 f 10.9 290.0 f 11.0 293.7 f 10.7 299.0 f 12.5 232.0 f 6.7 263.9 f 11.5 266.8 f 12.1 269.6 f 13.5 275.5 f 14.1 280.1 f 14.6 288.2 f 15.7 293.2 f 15.6 297.5 f 16.3 302.4 f 16.4 232.8 f 6.9 262.4 f 13.0 262.9 f 15.3 265.1 f 16.5 268.5 f 16.7 273.0 f 15.0 279.0 f 16.7 285.3 f 15.9 290.2 f 15.4 294.8 f 15.5 PROTOCOL 4 1 8 - 0 2 3 : ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBVTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T - 7 4 8 5 . 1 2 ) TABLE 3 (PAGE 2 ) : MATERNAL BODY WEIGHTS - SUMMARY DAY 1 7 MEANfS . D . 334.0 f 15.9 332.4 f 14.6 333.8 f 19.5 322.8 f 18.9 DAY 1 8 MEANfS.D . 350.8 f 1 6 . 3 344.9 f 14.9 347.8 * 19.9 336.5 f 21.4 DAY 1 9 MEANfS .D . 364.3 f 19.7 358.7 * 15.6 364.0 f 2 2 . 3 [ 21lb 353.0 f 22.4 DAY 20 MEANkS.D . 386.0 +_ 2 0 . 9 375.9 f 17.1 380.8 f 22.8 [ 21lb 368.5 f 22.2** DAY 2 1 MEANfS .D . 414.0 f 24.7 401.7 f 1 9 . 9 402.0 f 25.9 [ 2llb 391.2 f 23.6** [ 21lb . GRAVID UTERINE w WEIGHTS (G) MEANfS .D 108.5 f 18.6 104.9 f 15.3 4 [ 231c.d DAY 21C e MEANkS .D, 304.4 f 17.0 296.8 f 18.2 101.3 f 1 1 . 8 [ 231c 3 0 0 . 1 +_ 2 0 . 9 9 9 . 1 f 11.6 [ 20ld 291.5 19.5 [ 23lc.d [ 231c [ 20ld ________________________________________-------------------------------------------------------------------------------------------- DAY = DAY OF GESTATION [ 1 = NUMBER OF VALUES AVERAGED a. Dosage occurred on days 6 through 2 0 of gestation. b. Excludes values for dam 1 9 1 9 8 which was found dead on gestation day 1 7 . c. Excludes values from dams that delivered on day 2 1 of gestation. d . Excludes values for dams with no gravid uterine weight recorded. e. 2 1 C = Corrected maternal body weight (day 2 1 of gestation body weight minus the gravid uterine weight). * * Significantly different from the vehicle control group value ( ~ ~ 0 . 0 1 ) . PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 4 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - SUMMARY MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 6 MEANkS.D . +31.1 f 6.4 + 3 0 . 3 f 7.4 +31.8 f 9.9 +29.6 f 10.9 DAYS 6 - 9 MEAN_+.SD . +11.0 f 4.2 +10.1 f 4.7 +11.6 f 7.0 +6.2 f 8 . 8 * DAYS 9 - 12 MEANkS .D. +18.2 f 3.5 +16.6 f 5.0 +17.8 f 5.1 +16.8 7.6 DAYS 12 - 15 MEANkS .D . + 2 0 . 3 f 4.7 +19.0 f 5.4 +18.2 f 5.4 +16.9 f 7.4 w DAYS 15 - 18 MEANkS.D . +38.6 f 7.1 +36.0 f 6.8 + 3 6 . 3 f 5.0 +33.[ 62fl:lb8 . 8 00 DAYS 18 - 21 MEANfS.D . +63.2 f 11.5 +56.8 +- 8.2* +54.2 f 10.4** +54.7 f 9.0** DAYS 6 - 21 MEANfS .D. +151.3f 18.8 +138.5f 15.2* +138.2+ 20 . O f 21lb +128.4f 14.4** DAYS 0 - 21 MEANiS .D . +182.4+ 22.5 +168.8+ 18.7 +170.0f 26.0 [ 21lb +158.0* 22.5** DAYS DAYS 6 - 21C e 0 - 21C e MEAN_+S.D . MEANfS .D. +42.4 f 13.7 [ 231c.d +73.6 f 15.7 +33.6 f 15.6 +63.9 f 18.4 +36.5 f 15.3 [ 231c +6?.9 f 20.6 [ 21lb +28.9 f 11.6** I 20ld +58.5 f 19.1 [ 23lc.d [ 231c [ 20ld __________________._____________________----------------------- DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Dosage occurred on days 6 through 20 of gestation. b. Excludes values for dam 19198 which was found dead on gestation day 17. c. Excludes values from dams that delivered on day 21 of gestation. d. Excludes values for dams with no gravid uterine weight recorded. e. 21C = Corrected maternal body weight (day 21 of gestation body weight minus the gravid uterine weight). Significantly different from the vehicle control group value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the vehicle control group value (p<O.Ol). PROTOCOL 4 1 8 - 0 2 3 : ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T - 7 4 8 5 . 1 2 ) TABLE 5 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - SUMMARY PREGNANT N 25 22 24 22 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 6 DAYS 6 - 9 DAYS 9 - 1 2 MEANfS .D. MEAN+S.D. MEANtS .D. 22.1 f 2.5 22.5 f 1.8 23.0 f 2.3 22.1 f 2.1 22.4 f 2.2 22.6 f 1.2 23.0 f 2.6 23.4 f 3 . 1 [ 23lb 23.7 f 3.2 22.2 f 2.2 21.1 f 4.0 21.2 f 3.0 DAYS 1 2 - 1 5 MEANfS .D . 24.5 f 2.4 24.0 f 1 . 9 24.3 f 2.4 22.9 f 2.3 DAYS 1 5 - 18 DAYS 1 8 - 2 1 MEANfS .D . MEAN+S.D. 25.6 f 2.8 26.9 f 3.4 26.0 f 2 . 1 [ 21lb 2 4 . 9 f. 3.0* 26.1 f 24.8 f 2.8 2.9* 24.4 f 3.2 I 2llc 23.6 f 2.6**. [ 21lc DAYS 6 - 2 1 MEANfS .D . 24.5 f 2.0 24.0 f 1.6 24.4 f 2 . 5 2 2 . 1 f 2.0** DAYS 0 - 2 1 MEANfS .D . 24.0 f 1 . 9 23.6 f 1.5 24.0 f 2.4 I 2llc 22.6 f 2.0 I 21lc ________________--______________________-------------------------------------------------------------------------------------------- DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Dosage occurred on days 6 through 2 0 of gestation. b. Excludes values that were associated with spillage. c. Excludes values for dam 1 9 1 9 8 which was found dead on gestation day 17. Significantly different from the vehicle control group value (~50.05). * * Significantly different from the vehicle control group value ( ~ ~ 0 . 0 1 ) . PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSILJM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 6 (PAGE 1) : MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - SUMMARY -------._________.._____ GROUP DOSAGE (MG/KG/DAY)a --------____-__-.__-____ RATS TESTED 25 25 25 25 PREGNANT N 25 22 24 22 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 6 MEANfS .D. 91.8 f 8.7 91.6 f 8.3 92.5 f 9.6 89.6 f 7.2 DAYS 6 - 9 DAYS 9 - 12 MEANfS.D. MEANfS .D. 83.1 f 5.6 81.2 f 6.6 83.5 f 80.3 f 7.2 4.2 86.5 f 10.4 [ 23lb 83.3 f 1.7 79.4 f 12.4 76.7 f 8.3* DAYS 12 - 15 MEANfS .D. 81.5 f 6.3 80.7 _+ 5.3 80.4 f 5.5 78.2 f 6.5 P DAYS 15 - 18 MEAN+S,D. 77.7 f 8.1 79.6 f 5.1 19.2 f 6.0 16.6 f 8.8 0 [ 21lb [ 21lc . . DAYS 18 - 21 MEANiS D 71.0 f 8.0 61.2 f 6.7 66.2 f 5.8* 65.2 f 6.5** . r DAYS 6 - 21 MEANiS D. 78.2 f 5.0 11.3 f 3.8 78.1 f 4.6 21lc 14.5 f 4.2** DAYS 0 - 21 MEANfS . D. 77.8 f 4.5 71.3 f 3.5 17.9 & 4.3 * [ 21lc 75.1 4.0 [ 21lc _______________--___i___________________-----.------------------- DAYS = DAYS OF GESTATION [ ] = NUMBER OF VALUES AVERAGED a. Dosage occurred on days 6 through 20 of gestation. b. Excludes values that were associated with spillage. c. Excludes values for dam 19198 which was found dead on gestation day 17. * Significantly different from the vehicle control group value ( ~ ~ 0 . 0 5 ) . * * Significantly different from the vehicle control group value (p~O.01). b PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 7 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY PREGNANT FOUND DEAD DELIVERED RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION CORPORA LUTEA IMPLANTATIONS LITTER SIZES N MEANfS.D . MEANfS .D. MEANfS.D . LIVE FETUSES N MEANtS .D. DEAD FETUSES N RESORPTIONS EARLY RESORPTIONS MEANfS.D. N MEANiS .D. LATE RESORPTIONS N DAMS WITH ANY RESORPTIONS N(%) DAMS WITH ALL CONCEPTUSES RESORBED N(%) DAMS WITH VIABLE FETUSES N($) ZS(100.0) O( 0.0) 1( 4.0) 24 18.6 f 3.2 15.3 f 2.8 14.9 f 2.9 358 14.9 f 2.9 0 0.4 f 0.7 10 0.4 f 0.7 0 7 ( 29.2) 0 24(100.0) 22( 88.0) O ( 0.0) O( 0.0) 22 17.9 f 3.2 15.2 f 2.3 14.4 f 2.2 316 14.4 f 2.2 0 0.8 f 1.1 18 0.8 f 1.1 0 10( 45.4) 0 22(100.0) ' 24( 96.0) O ( 0.0) 1( 4.2) 23 15.9 f 2.4** 15.0 f 1.9 14.4 f 2.2 331 14.4 f 2.2 0 0.6 f 0.8 15 0.6 f 0 . 8 0 12( 52.2) 0 23 (100.0) 22( 88.0) 1( 4.5) O ( 0.0) 21 17.6 f 2.4 15.7 f 1.7 14.8 f 1.7 311 14.8 f 1.7 0 0.8 f 1.1 18 0.8 f 1.1 0 10( 47.6) 0 21(100.0) PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 8 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVEREDFETUSES) - SUMMARY GROUP DOSAGE (MG/KG/DAY)a -------------___---_____________________- LITTERS WITH ONE OR MORE LIVE FETUSES N 24 IMPLANTATIONS MEANfS . D. 15.3 f 2.8 LIVE FETUSES N MEANfS.D . 358 14.9 f 2.9 LIVE MALE FETUSES N 189 22 15.2 f 2.3 3 16 14.4 i: 2.2 154 23 15.0 f 1.9 331 14.4 f 2.2 161 21 15.7 f 311 14.8 f 156 1.7 1.7 % LIVE MALE FETUSES/LITTER MEANfS.D . 52.4 f 8.9 49.0 f 12.1 48.2 f 13.4 50.5 f 12.7 LIVE FETAL BODY WEIGHTS (GRAMS)/LITTER MEANfS.D. R MALE FETUSES MEANfS .D. FEMALE FETUSES MEAN+S.D . 5.34 f 0.39 5.48 f 0.39 5.18 f 0.41 5.36 f 0.31 5.54 f 0.35 5.21 f 0.32 5.17 f 0.30 5.30 f 0.29 5.06 f 0.32 4.87 f 0.20** 5.00 f 0.22** 4.73 f 0.21** * % RESORBED CONCEPTUSES/LITTER MEANfS.D. 2.8 f 4.9 5.3 8.1 4.5 f 5.9 5.3 f 6.7 ________________________________________-------------------------------------------------------------------------------------------- a . Dosage occurred on days 6 through 20 of gestation. * * Significantly different from the vehicle control group value (peO.01). b PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBTANE SULFONATE (PFBS) TABLE 9 (PAGE 1): FETAL ALTERATIONS - SUMMARY LITTERS EVALUATED FETUSES EVALUATED LIVE LITTERS WITH FETUSES WITH ANY ALTERATION OBSERVED FETUSES W I T H ANY ALTERATION OBSERVED N N N N(%) N(%) 24 358 358 4 ( 16.7) 4( 1.1) 22 316 316 8 ( 36.4) 13 ( 4.1)* * 23 331 331 3 ( 13.0) 3 ( 0.9) 21 311 311 3 ( 14.3) 3 ( 1.0) w P PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 10 (PAGE 1): FETAL GROSS EXTERNAL ALTERATIONS - SUMMARY PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) ' TABLE 11 (PAGE 1) : FETAL SOFT TISSUE ALTERATIONS - SUMMARY EYES: FOLDED RETINA LITTER INCIDENCE FETAL INCIDENCE N(%) N(%) O ( 0.0) O( 0.0) 1( 4 . 5 ) 1( 0.7) O( 0.0) O( 0.0) EYES: ABSENT EYE LITTER INCIDENCE FETAL INCIDENCE N(%) N(%) 1( 4.2) 1( 0.6) O ( 0.0) O ( 0.0) O( 0.0) O ( 0.0) VESSELS: UMBILICAL ARTERY DESCENDED TO THE LEFT OF THE URINARY BLADDER LITTER INCIDENCE N(%) 1( 4.2) 2( 9.1) FETAL INCIDENCE N(%) 1( 0.6) 2( 1.3) O( 0.0) O( 0.0) R VESSELS: INNOMINATE ARTERY ABSENT LITTER INCIDENCE N(%) O( 0.0) O( 0.0) FETAL INCIDENCE N(%) O( 0.0) O ( 0.0) --__---______-__________________________------------------------------------------ a. Dosage occurred on days 6 through 20 of gestation. O( 0.0) O( 0.0) O( 0.0) O( 0.0) O ( 0.0) O( 0.0) I 8 w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 12 (PAGE 1) ; FETAL SKELETAL ALTERATIONS - SUMMARY (See f o o t n o t e s on t h e l a s t page of t h i s t a b l e . ) THORACIC VERTEBRAE: CENTRUM, BIFID LITTER INCIDENCE N(%) FETAL INCIDENCE N(%) 2 ( 8.3) 2( l . l ) b THORACIC VERTEBRAE: ARCHES, FUSED LITTER INCIDENCE N(%) FETAL INCIDENCE N(0) THORACIC VERTEBRAE: HEMIVERTEBRA LITTER INCIDENCE N(%) FETAL INCIDENCE N(%) 1( 4.2) 1( 0 . 5 ) b THORACIC VERTEBRAE: CENTRUM, UNILATERAL OSSIFICATION LITTER INCIDENCE N(%) 1( 4.2) FETAL INCIDENCE N(%) 1( 0 . 5 ) b THORACIC VERTEBRAE: CENTRUM, NOT OSSIFIED LITTER INCIDENCE N(%) FETAL INCIDENCE N(%) 1( 4.2) 1( 0 . 5 ) b 4 ( 18.2) 4( 2.4)c O( 0.0) O( 0.0) O( 0 . 0 ) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) 3 ( 13.0) 3( 1.7) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O ( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) O( 0.0) PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 12 (PAGE 2) : FETAL SKELETAL ALTERATIONS - SUMMARY LITTERS EVALUATED N 24 22 23 21 FETUSES EVALUATED N 185 166 174 161 LIVE N 185 166 174 161 ---------_______________________________-------------------------------------------------------------------------------------------- STERNAL CENTRA: SUMMARIZATION (Includes fused and asymmetric sternal centra): LITTER INCIDENCE FETAL INCIDENCE N(%) N(%) O( 0.0) O ( 0.0) 1( 4.5) 1( 0 . 6 ) O( 0.0) O( 0.0) O( 0.0) O( 0.0) STERNAL CENTRA: FUSED LITTER INCIDENCE FETAL INCIDENCE N(%) N(%) O ( 0.0) O( 0.0) 1( 4.5) 1( 0 . 6 ) d O( 0.0) O( 0.0) O( 0.0) O( 0.0) STERNAL CENTRA: ASYMMETRIC % LITTER INCIDENCE N(%) O( 0.0) FETAL INCIDENCE N(%) O ( 0.0) P 4 PELVIS: PUBIS, INCOMPLETELY OSSIFIED O( 0.0) O( 0.0) O( 0.0) O( 0.0) LITTER INCIDENCE N(%) O ( 0.0) 1( 4.5) O( 0.0) O( 0.0) FETAL INCIDENCE N(%) O( 0.0) 1( 0.6) O( 0.0) O( 0.0) -_-___-_______-_________________________-------------------------------------------------------------------------------------------- a . Dosage occurred on days 6 through 20 of gestation. b. Fetus 19120-5 had other skeletal alterations. c . Fetus 19138-9 had other skeletal alterations. d. Fetus 19140-12 had other skeletal alterations. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 13 (PAGE I): FETAL OSSIFICATION SITES - CAESAREAN-DELIVERED LIVE FETUSES (DAY 21 OF GESTATION) - SUMMARY HYOID MEANfS.D . 1.00 f 0.00 VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS (PAIRS) MEANfS .D. MEANfS . D. MEANfS.D . MEANfS .D. MEANfS . D. MEANfS.D . 7.00 f 13.08 f 5.92 f 3.00 f 1.75 f 0.00 0.13 0.13 0.00 0.92 13.05 f 0 . 0 8 STERNUM MANUBRIUM STERNAL CENTERS XIPHOID MEAN+S .D. MEANfS.D . MEANfS.D . 1.00 f 3.98 f 1.00 f 0.00 0.06 0.00 FORELIMB b CARPALS METACARPALS DIGITS PHALANGES MEANfS.D . MEANfS.D . MEANfS .D. MEANfS.D. 0.00 f 4.00 f 5.00 f 8.37 f 0.00 0.02 0.00 0.74 HINDLIMB b TARSALS METATARSALS DIGITS PHALANGES MEANfS.D. MEANfS.D . MEANfS .D . MEANfS . D. 0.04 f 4.85 f 5.00 f 6.57 f 0.16 0.21 0.00 1.27 a. Dosage occurred on days 6 through 20 of gestation. b. Calculated as average per limb. 1.00 f 0.00 7.00 f 13.03 f 5.95 f 3.00 f 7.64 f 0.00 0.06 0.08 0.00 0.90 13.02 f 0.04 1.00 f 3.98 f 1.00 f 0.00 0.07 0.00 0.00 f 3.97 f 5.00 f 8.35 f 0.00 0.09 0.00 0.71 0.03 f 4.80 f 5.00 f 6.52 f 0.08 0.27 0.00 1.11 0.99 f 0.04 0.98 f 0.05 7.00 f 13.07 f 5.92 f 3.00 f 7.65 f 0.00 0.12 0.12 0.00 0.70 13.06 f 0.10 7.00 f 13.08 f 5.91 f 3.00 f 7.10 f 0.00 0.17 0.17 0.00 0.58 13.06 f 0.11 1.00 f 4.00 f 1.00 f 0.00 0.02 0.00 1.00 f 3.98 f 1.00 f 0.00 0.05 0.00 0.00 f 4.00 f 5.00 f 8.37 f 0.00 0.00 0.00 0.64 0.00 f 4.00 f 5.00 f 8.27 f 0.00 0.00 0.00 0.64 0.02 f 0.06 4.81 f 0.28 5.00 f 0.00 6.27 f 1.10 _--__--__--__-___ 0.00 f 0.00 4.60 f 0.31** 5.00 f 0.00 5.65 f 0.72** _--__-___-___--__-___ P c 03 PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBFANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 14 (PAGE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA 19101 NO ADVERSE FINDINGS 19102 NO ADVERSE FINDINGS 19103 DG( 14- 19) LOCALIZED ALOPECIA: LIMBS 19104 NO ADVERSE FINDINGS 19105 NO ADVERSE FINDINGS 19106 NO ADVERSE FINDINGS 19107 NO ADVERSE FINDINGS 19108 NO ADVERSE FINDINGS 19109 DG( 9 - 21) LOCALIZED ALOPECIA: LIMBS a 19110 19111 DG( 21 1 NO ADVERSE FINDINGS DELIVERED AND SACRIFICED 19112 NO ADVERSE FINDINGS 19113 NO ADVERSE FINDINGS 19114 NO ADVERSE FINDINGS 19115 DG( 15- 21) LOCALIZED ALOPECIA: LIMBS a 19116 NO ADVERSE FINDINGS 19117 NO ADVERSE FINDINGS 19118 NO ADVERSE FINDINGS 19119 NO ADVERSE FINDINGS 19120 NO ADVERSE FINDINGS 19121 NO ADVERSE FINDINGS 19122 NO ADVERSE FINDINGS 19123 NO ADVERSE FINDINGS 19124 NO ADVERSE FINDINGS 19125 NO ADVERSE FINDINGS .................................................................................................................................... DG = DAY O F PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 4 1 8 - 0 2 3 : ORAL (GAVAGE) DEVELOPMENTAL T O X I C I T Y STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE ( P F B S ) I N RATS ( S P O N S O R ' S STUDY NUMBER: T - 7 4 8 5 . 1 2 ) TABLE 1 4 (PAGE 2 ) : C L I N I C A L OBSERVATIONS - INDIVIDUAL DATA .................................................................................................................................... GROUP I1 LOW DOSAGE 100 MG/KG/DAY .................................................................................................................................... FAT # DESCRIPTION .................................................................................................................................... 19126 19127 19128 NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS 19129 NO ADVERSE FINDINGS 19130 NO ADVERSE FINDINGS 19131 NO ADVERSE FINDINGS 19132 NO ADVERSE FINDINGS 19133 NO ADVERSE F I N D I N G S 19134 NO ADVERSE FINDINGS 19135 NO ADVERSE FINDINGS 19136 NO ADVERSE FINDINGS 19137 NO ADVERSE FINDINGS 19138 NO ADVERSE FINDINGS 19139 NO ADVERSE FINDINGS 19140 NO ADVERSE FINDINGS 19141 NO ADVERSE FINDINGS 19142 NO ADVERSE FINDINGS 19143 NO ADVERSE FINDINGS 19144 NO ADVERSE FINDINGS 19145 NO ADVERSE FINDINGS 19146 NO ADVERSE FINDINGS 19147 NO ADVERSE FINDINGS 19148 DG( 15- 21) LOCALIZED ALOPECIA: LIMBS a 19149 NO ADVERSE FINDINGS 19150 NO ADVERSE FINDINGS .................................................................................................................................... DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. 0 wh) PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 14 (PAGE 3 ) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA ----------.-_-_-___-____________________------------------------------- GROUP I11 MIDDLE DOSAGE 300 MG/KG/DAY --------.______-________________________---------------------------- RAT # DESCRIPTION .................................................................................................................................... 19151 NO ADVERSE FINDINGS 19152 NO ADVERSE FINDINGS 19153 NO ADVERSE FINDINGS 19154 NO ADVERSE FINDINGS 19155 NO ADVERSE FINDINGS 19156 NO ADVERSE FINDINGS 19157 DG( 21 ) DELIVERED AND SACRIFICED 19158 NO ADVERSE FINDINGS 19159 NO ADVERSE FINDINGS 19160 NO ADVERSE FINDINGS 19161 NO ADVERSE FINDINGS 19162 NO ADVERSE FINDINGS 19163 NO ADVERSE FINDINGS 19164 NO ADVERSE FINDINGS 19165 NO ADVERSE FINDINGS 19166 NO ADVERSE FINDINGS 19167 NO ADVERSE FINDINGS 19168 NO ADVERSE FINDINGS 19169 NO ADVERSE FINDINGS 19170 NO ADVERSE FINDINGS 19171 NO ADVERSE FINDINGS 19172 NO ADVERSE FINDINGS 19173 DG( 7- 21) LOCALIZED ALOPECIA: LIMBS a 19174 NO ADVERSE FINDINGS 19175 NO ADVERSE FINDINGS -----_---___--_------------------------------.-------------------------------- DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 14 (PAGE 4 ) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA 19176 19177 19178 19179 19180 19181 19182 19183 19184 19185 19186 19187 19188 19189 19190 19191 19192 ' 19193 19194 19195 19196 19197 19198 19199 19200 DG( 1 7 ) DG( 17 ) DG( 18 1 DG( 17 ) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS COLD TO TOUCH DEHYDRATION a FOUND DEAD NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS FOUND DEAD NO ADVERSE FINDINGS NO ADVERSE FINDINGS R w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 15 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA GROUP DOSAGE (MG/KG/DAY) RAT NUMBER ._ DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a _________.____-_____--------------- 19101 19102 19103 19104 19105 19106 19107 19108 19109 19110 DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 19111 DG 21 P 15 DELIVERED ON DAY 21 OF GESTATION. UTERINE CONTENTS: 16 IMPLANTATION SITES ( 2 DELIVERED PUPS, 13 FETUSES AND 1 EARLY RESORPTION IN UTERO) .b ALL TISSUES APPEARED NORMAL. 19112 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19113 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19114 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19115 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19116 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19117 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19118 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19119 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19120 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19121 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19122 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19123 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19124 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19125 DG 21 P 15 ALL TISSUES APPEARED NORMAL. ________________________________________---------------.------ DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 14) for external observations confirmed at necropsy. b. Refer to table 20 for gross, soft tissue and skeletal examinations. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 15 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA ----r----__________.____________________~~~~------------------------------------------------------------------------------------~--- GROUP RAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a .................................................................................................................................... I1 100 19126 DG 21 P 15 ALL TISSUES APPE!ARED NORMAL. 19127 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19128 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19129 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19130 19131 DG 21 P DG 21 NP 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 19132 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19133 19134 DG 21 P DG 21 P 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 19135 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19136 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19137 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19138 DG 21 P 15 ALI, TISSUES APPEARED NORMAL. 19139 DG 21 P 15 ALL TISSUES APPEARED NORMAL. wl 19140 DG 21 P 15 ALL TISSUES APPEARED NORMAL. P 19141 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19142 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19143 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19144 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19145 DG 21 NP 15 ALL TISSUES APPEARED NORMAL. 19146 19147 19148 DG 21 P DG 21 P DG 21 NP 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 19149 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19150 DG 21 P 15 ALL TISSUES APPEARED NORMAL. ---__--____----__---____________________-----.~~-----~-------~---~------ DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observatlons table (Table 14) for external observations confirmed at necropsy. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 15 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA PREGNANCY DOSAGES STATUS ADMINISTERED ._ 19151 19152 19153 19154 19155 19156 DG 21 P DG 21 NP DG 21 P DG 21 P DG 21 P DG 21 P 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 19157 DG 21 P 15 DELIVERED ON DAY 21 OF GESTATION. UTERINE CONTENTS: 17 IMPLANTATION SITES (1 DELIVERED PUP AND 16 FETUSES IN UTERO).b ALL TISSUES APPEARED NORMAL. 19158 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19159 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19160 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19161 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19162 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19163 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19164 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19165 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19166 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19167 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19168 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19169 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19170 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19171 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19172 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19173 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19174 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19175 DG 21 P 15 ALL TISSUES APPEARED NORMAL. ____________-___________________________--.---.---------------------- DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 14) for external observations confirmed at necropsy. b. Refer to table 20 for gross, soft tissue and skeletal examinations. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 15 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA GROUP RAT DOSAGE (MG/KG/DAY) NUMBER .................................. IV 1000 19176 19177 19178 DG 21 P DG 21 P DG 21 P 19179 DG 21 P 19180 DG 21 P 15 ALL TISSUES APPE'ARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 19181 DG 18 NP 12 FOUND DEAD ON DAY 18 OF GESTATION. (DEATH OCCURRED PRIOR TO DOSAGE). ALL TISSUES APPEARED NORMAL. 19182 19183 19184 19185 19186 19187 19188 19189 19190 19191 19192 19193 19194 19195 19196 19197 DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 NP DG 21 NP DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P DG 21 P 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 15 ALL TISSUES APPEARED NORMAL. 19198 DG 17 P 12 FOUND DEAD ON DAY 17 OF GESTATION. (DEATH OCCURRED 7 MINUTES AFTER DOSAGE). ESOPHAGUS: PERFORATION (0.1CM IN DIAMETER). THORACIC CAVITY: RED SUBSTANCE ( 3 ML) UTERINE CONTENTS: 16 IMPLANTATION SITES (15 FETUSES AND 1 EARLY RESORPTION IN UTERO).b ALL OTHER TISSUES APPEARED NORMAL. 19199 DG 21 P 15 ALL TISSUES APPEARED NORMAL. 19200 DG 21 P 15 ALL TISSUES APPEARED NORMAL. __________-.__---__---------------.-----------.-------------------------------- DG = DAY OF PRESUMED GESTATION P = PREGNANT NP = NOT PREGNANT a. Refer to the individual clinical observations table (Table 14) for external observations confirmed at necropsy. b. Refer to table 20 for gross, soft tissue and skeletal examinations. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 16 (PAGE 1): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA 19101 P 224. 264. 267. 270. 273. 276. 283. 292. 294. 301. 311. 324. 342. 19102 P 237. 259. 260. 265. 269. 275. 281. 289. 291. 294. 305. 318. 330. 19103 P 228. 264. 264. 261. 271. 275. 283. 287. 297. 297. 307. 315. 326. 19104 P 223. 253. 258. 264. 271. 274. 282. 291. 290. 295. 308. 320. 332. 19105 P 232. 263. 265. 271. 274. 281. 284. 296. 298. 304. 313. 332. 349. 19106 P 228. 266. 270. 274. 279. 285. 290. 293. 297. 306. 312. 329. 337. 19107 P 240. 277. 279. 286. 288. 296. 303. 307. 314. 323. 333. 343. 355. 19108 P 231. 255. 256. 261. 263. 276. 281. 284. 290. 290. 303. 316. 318. 19109 P 221. 241. 246. 250. 252. 256. 263. 271. 273. 280. 287. 297. 306. 19110 P 236. 269. 273. 272. 279. 285. 290. 297, 301. 308. 318. 331. 343. 19111 P 242. 264. 266. 264. 273. 278. 284. 291. 293. 301. 307. 320. 329. 19112 P 224. 258. 267. 270. 271. 274. 282. 284. 291. 298. 305. 323. 333. 19113 P 238. 263. 268. 273. 280. 281. 291. 295. 300. 305. 312. 329. 341. 19114 P 230. 275. 280. 284. 288. 294. 305. 310. 317. 318. 329. 334. 352. 19115 P 231. 258. 268. 270. 276. 279. 284. 291. 296. 302. 311. 318. 333. 19116 P 220. 252. 258. 266. 269. 269. 278. 280. 280. 292. 307. 312. 316. 19117 P 238. 274. 281. 285. 290. 295. 302. 310. 317. 321. 333. 341. 350. 19118 P 226. 258. 255. 260. 263. 269, 275. 280. 284. 289. 302. 309. 312. 19119 P 228. 253. 257. 261. 261. 267. 273. 280. 287. 291. 303. 315. 315. 19120 P 234. 258. 261. 266. 266. 271. 270. 289. 286. 289. 302. 311. 320. 19121 P 232. 268. 277. 275. 281. 291. 294. 301. 307. 315. 326. 339. 354. 19122 P 232. 264. 264. 265. 270. 268. 277. 285. 287. 292. 296. 302. 316. 19123 P 243. 275: 278. 286. 289. 297. 300. 310. 325. 329. 343. 354. 371. 19124 P 240. 267. 267. 273. 269. 273. 286. 294. 296. 313. 316. 318. 332. 19125 P 234. 271. 271. 273. 279. 274. 288. - - - - - - _ _ 2_ 9_ 1_ ._ 293. _ _ 3_ 0_ 2_ ._ 3_ 1_ 6_ ._ 323. 339. ------- ----__ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). b PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 16 (PAGE 2): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA _______._________.______________________-----.---------------------- RAT # GROUP I VEHICLE 0 MG/KG/DAY __.___________.__.______________________-----.---------------------- PREGNANCY GRAVID STATUS DAY 18 19 20 21 UTERINE WEIGHT (G) --______________________________________-..---.--------------------- 19101 P 365. 378. 401. 429. 136.05 19102 P 344. 361. 378. 411. 106.33 19103 P 343. 353. 382. 412. 111.83 19104 P 347. 358. 379. 402. 89.53 19105 P 353. 375. 404. 431. 113.66 19106 P 353. 365. 382. 416. 108.74 19107 P 374. 379. 405. 449. a 19108 P 337. 347. 366. 402. 113.60 19109 P 320. 329. 345. 366. 87.54 19110 P 363. 386. 414. 444. 128.41 19111 P 343. 354. 374. 405 .b 108.44~ 19112 P 355. 366. 391. 420. 121.15 19113 P 357. 368. 385. 419. 106.85 19114 P 378. 387. 404. 446. 116.38 19115 P 353. 363. 388. 381. 117.87 19116 P 338. 360. 373. 403. 88.26 19117 P 359. 379. 398. 420. 94.09 19118 P 335. 320. 358. 387. 105.74 19119 P 330. 346. 363. 385. 87.37 19120 P 336. 354. 371. 398. 98.50 19121 P 373. 388. 414. 448. 133.60 19122 P 325. 339. 354. 376. 57.00 19123 P 383. 409. 433. 464. 129.47 19124 P 352. 370. 390. 417. 119.07 19125 P 355. 373. 399. 420. 123.61 __________-___-_---_____________________--.---------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Gravid uterine weight not recorded. b. Dam 19111 delivered on day 21 of gestation. c. Uterine weight taken including the two delivered pups (pups weighed outside the uterus); value was excluded from group averages and statistical analyses. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 16 (PAGE 3): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA 19126 P 19127 P 19128 P 220. 234. 221. 251. 269. 259. 258. 276. 263. 260. 281. 268. 257. 286. 277. 271. 285. 270. 19129 P 224. 258. 259. 265. 272. 277. 19130 P 230. 250. 251. 256. 259. 261. 19131 NP 228. 231. 233. 234. 228. 236. 19132 P 19133 P 232. 232. 270. 260. 267. 265. 258. 270. 267. 271. 272. 279. 19134 P 238. 255. 257. 263. 263. 270. 19135 P 230. 252. 256. 263. 269. 274. 19136 P 19137 P 233. 241. 276. 277. 278. 267. 288. 270. 286. 280. 296. 283. 19138 P 240. 264. 268. 269. 272. 275. 19139 P 232. 261. 266. 267. 270. 275. 19140 P 19141 P 19142 P 236. 242. 236. 264. 284. 258. 266. 287. 262. 269. 290. 263. 272. 297. 269. 274. 303. 271. 19143 P 224. 255. 260. 262. 269. 272. 19144 P 237. 265. 266. 273. 274. 278. 19145 NP 228. 241. 244. 243. 238. 244. 19146 P 225. 255. 260. 262. 267. 273. 19147 P 239. 276. 281. 284. 283. 291. 19148 NP 224. 248. 255. 248. 251. 255. 19149 P 235. 254. 252. 254. 265. 265. 19150 P 243. 278. 277. 282. 288. 288. .................................................................... P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERnGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 274. 294. 282. 285. 264. 238. 280. 284. 271. 281. 302. 292. 282. 283. 287. 305. 275. 273. 283. 249. 275. 297. 258. 270. 299. 280. 297. 282. 290. 277. 239. 289. 295. 284. 285. 305. 300. 289. 283. 290. 319. 284. 277. 290. 250. 283. 304. 248. 274. .3- 0- -2-.- - - 293. 306. 290. 296. 278. 233. 292. 296. 282. 290. 310. 303. 293. 292. 298. 319. 283. 281. 288. 244. 289. 306. 246. 276. 3_ 0_ 0_ _. 298. 309. 286. 305. 279. 241. 294. 302. 291. 295. 319. 306. 301. 296. 302. 328. 284. 286. 295. 248. 291. 313. 245. 283. 314. 308. 324. 301. 310. 286 .' 245. 310. 311. 301. 305. 331. 314. 308. 303. 314. 337. 298. 299. 296. 251. 301. 318. 250. 295. 326. 315. 330. 314. 324. 298, 244. 320. 326. 318. 312. 341. 329. 313. 310. 324. 355. 305. 303. 307. 244. 308. 326. 245. 297. 336. 330. 347. 326. 338. 312. 238. 330. 336. 330. 332. 358. 349. 333. 318. 338. 355. 309. 316. 326. 246. 317. 346. 247. 314. 353. P c 90 R w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 16 (PAGE 4): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA 19126 P 349. 362. 382. 411. 19127 P 356. 376. 394. 424. 19128 P 339. 349. 367. 390. 19129 P 345. 361. 372. 407. 19130 P 315. 329. 343. 369. 19131 NP 245. 250. 244. 246. 19132 P 345. 354. 375. 398. 19133 P 353. 366. 385. 421. 19134 P 349. 361. 392. 411. 19135 P 332. 359. 364. 398. 19136 P 366. 382. 406. 420. 19137 P 365. 373. 388. 415. 19138 P 342. 356. 368. 392. m 19139 P 328. 334. 350. 364. 0 19140 P 352. 366. 375. 406. 19141 P 369. 382. 403. 436. 19142 P 328. 346. 358. 382. 19143 P 334. 348. 365. 393. 19144 P 342. 358. 375. 403. 19145 NP 247. 247. 252. 253. 19146 P 331. 342. 360. 384. 19147 P 357. 370. 389. 421. 19148 NP 251. 248. 250. 254. 19149 P 325. 335. 360. 368. 19150 P 366. 382. 398. 425. 101.40 109.77 97.44 86.13 9- 2- .- 2- 0- 127.44 113.72 117.40 110.21 112.53 113.84 86.10 61.36 111.03 93.85 102.05 117.48 117.92 --_-- 91.47 11_3_.4-3- - 107.03 124.49 P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (GI. PROTOCOL 4 1 8 - 0 2 3 : ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 16 (PAGE 5 ) : MATERNAL BODY WEIGHTS - INDIVIDUAL DATA PREGNANCY STATUS DAY 0 6 7 8 9 10 11 12 19151 P 224. 249. 252. 252. 251. 265. 274. 276. 19152 NP 225. 252. 254. 254. 256, 260. 263. 262. 19153 P 222. 268. 270. 276. 273. 282. 288. 288. 19154 P 243. 271. 268. 263. 266. 270. 281. 283. 19155 P 234. 266. 270. 215. 276. 287. 293. 305. 19156 P 242. 262. 268. 263. 270. 278. 284. 288. 19157 P 228. 269. 272. 270. 278. 280. 290. 294. 19158 P 238. 280. 288. 290. 294. 299. 310. 316. 19159 P 236. 262. 262. 270. 270. 278. 285. 292. 19160 P 232. 267. 260. 265. 271. 270. 283. 286. 19161 P 230. 268. 276. 281. 291. 296. 312. 316. 19162 P 223. 243. 252. 252. 258. 264. 265. 274. 19163 P 220. 251. 250. 254. 264. 268. 274. 279. 19164 P 229. 280. ' 286. 292. 301. 307. 314. 321. 19165 P 228. 260. 267. 270. 274. 282. 289. 297. 19166 P 19167 P 232. 228. 256. 257. 262. 265. 265. 268. 273. 273. 278. 277. 285. 288. 288. 292. 19168 P 241. 271. 266. 272. 280. 280. 287. 290. 19169 P 239. 275. 278. 284. 292. 295. 303. 308. 19170 P 224. 248. 242. 247. 253. 260. 260. 268. 19171 P 235. 245. 258. 262. 269. 261. 276. 277. 19172 P 233. 284. 292. 299. 304. 315. 322. 327. 19173 P 240. 278. 276. 280. 292. 292. 303. 302. 19174 P 231. 256. 258. 252. 262. 264. 270. 279. 19175 P 237. 267. 264. 269. 277. 275. 282. 292. - - _ _ _ _ _ _ _ _ - - _ _ _ - - - _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - --------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . 13 281. 255. 298. 284. 310. 294. 295. 320. 295. 290. 319. 275. 278. 322. 299. 297. 302. 294. 315. 272. 291. 337. 303. 281. 288. 14 288. 254. 305. 290. 316. 298. 302. 329. 301. 294. 329. 279. 288. 322. 298. 299. 307. 299. 320. 275. 284. 338. 308. 287. 301. 300. 255. 318. 297. 318. 308. 311. 345. 309. 300. 339. 285. 290. 331. 313. 309. 313. 312. 331. 284. 294. 349. 317. 291. 311. 308. 252. 327. 304. 332. 316. 324. 360. 319, 314. 350. 302. 298. 348. 320. 318. 332. 319. 347. 294. 304. 355. 323. 298. 319. 322. 253. 345. 315. 346. 325. 337. 370. 329. 326. 364. 306. 305. 355. 328. 336. 335. 330. 358. 304. 323. 371. 339. 313. 329. w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 16 (PAGE 6): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA 19151 P 19152 NP 335. 252. 349. 257. 367. 259. 391. 263. 9_ 9_ ._ 2_ 4_ 19153 P 352. 37s. 392. 436. 109.01 19154 P 320. 337. 352. 367. 90.83 19155 P 358. 381. 394. 417. 120.07 19156 P 19157 P 338. 354. 349. 367. 364. 386. 388. 417.a 74.51 113.46b 19158 P 388. 409. 417. 437. 98.49 19159 P 344. 361. 385. 389. 101.74 19160 P 342. 362. 379. 397. 115.40 19161 P 370. 387. 400. 430. 98.97 19162 P 320. 330. 347. 366. 90.93 19163 P 324. 337. 353. 376. 94.62 19164 P 369. 388. 409. 421. 108.14 19165 P 347. 362. 386. 395. 103.27 19166 P 348. 365. 393. 413. 114.38 19167 P 353. 374. 390. 413. 112.35 19168 P 354. 376. 385. 413. 110.86 19169 P 366. 384. 404. 438. 115.11 19170 P 316. 331. 339. 361. 93.25 19171 P 337. 345. 371. 385. 97.65 19172 P 391. 403. 424. 450. 107.27 19173 P 350. 367. 374. 386. 86.25 19174 P 323. 330. 344. 363. 79.82 19175 P 348. 367. 383. 400. 107.15 ___________-___-_--_____________________-------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). b a. Dam 19157 delivered on day 21 of gestation. b. Uterine weight taken including the one delivered pup (pup weighed outside the uterus); value was excluded from group averages and statistical analyses. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 16 (PAGE 7): MATERNAL BODY WEIGHTS - INDIVIDUAL DATA 19176 P 233. 265. 266. 266. 272. 278. 282. 285. 291. 298. 302. 315. 327. 19177 P 242. 278. 282. 284. 290. 285. 286. 293. 307. 312. 321. 332. 353. 19178 P 222. 238. 241. 242. 250. 257. 242. 254. 262. 266. 276. 285. 286. 19179 P 228. 260. 264. 268. 272. 280. 269. 279. 281. 290. 301. 310. 328. 19180 P 220. 257. 264. 266. 271. 272. 270. 277. 283. 294. 302. 308. 320. 19181 NP 232. 262. 256. 246. 244. 251. 256. 254. 253. 259. 253. 261. 242. 19182 P 236. 279. 286. 294. 289. 300. 308. 316. 317. 317. 323. 338. 355. 19183 P 237. 267. 267. 275. 279. 283. 290. 294. 301. 306. 316. 323. 336. 19184 P 237. 275. 276. 281. 289. 282. 300. 301. 304. 317. 322. 333. 344. 19185 P 231. 268. 266. 274. 265. 268. 278. 287. 295. 301. 309. 328. 325. 19186 P 221. 242. 251. 250. 255. 268. 272. 277. 273. 280. 282. 294. 308. 19187 NP 224. 231. 234. 230. 232. 233. 232. 231. 234. 232. 233. 236. 236. 3 19188 NP 226. 258. 261. 262. 263. 266. 271. 269. 268. 265. 262. 266. 262. m 19189 P 229. 264. 269. 274. 284. 284. 295. 305. 308. 295. 306. 316. 318. w 19190 P 228. 268. 267. 274. 272. 278. 290. 300. 302. 312. 312. 324. 339. 19191 P 232. 264. 262. 256. 253. 256. 267. 271. 277. 277. 290. 304. 315. 19192 P 238. 255. 250. 248. 246. 246. 257. 263. 272. 275. 280. 291. 302. 19193 P 240. 270. 261. 269. 273. 281. 283. 291. 290. 295. 299. 308. 315. 19194 P 231. 256. 258. 255. 261. 271. 271. 280. 282. 288. 298. 307. 321. 19195 P 234. 260. 266. 245. 255. 257. 264. 273. 285. 285. 292. 298. 308. 19196 P 241. 268. 272. 274. 277. 285. 290. 292. 296. 299. 312. 325. 334. 19197 P 239. 232. 234. 235. 234. 245. 263. 265. 266. 276. 275. 288. 292. 19198 P 224. 254. 225. 239. 245. 254. 262. 270. 279. 279. 288. 296. 306. 19199 P 234. 266. 272. 217. 283. 281. 294. 296. 300. 303. 306. 318. 318. _ 19200 P ___ _ _ 244. __ _ 286. ___ 285. ___ _ 286. __ _ 293. -_ _ 295. ___ 304. ___ _ 308. __ _ _ 3_ _1 _3_ _. _ _ 321. __ _ 337. __- - . 331. --- - .- 352. - - - -- --- - - - - - - - - - - - - - P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (GI. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASS'IUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 16 (PAGE 8) : MATERNAL BODY WEIGHTS - INDIVIDUAL DATA ---------____..__--_____________________.~~--..----~----.-------------- RAT # GROUP IV HIGH DOSAGE 1000 MG/KG/DAY -----_--_________--_____________________--~--.----------------------------- PREGNANCY GRAVID STATUS DAY 18 19 20 21 UTERINE WEIGHT (G) -----------_____-_--____________________--~-------------------~-----------------------~---~------~------------------------~-~--~---- 19176 P 19177 P 19178 P 19179 P 19180 P 19181 NP 340. 354. 373. 395. 370. 379. 401. 432. 301. 316. 330. 350. 340. 361. 372. 400. 336. 353. 363. 390. FOUND DEAD ON DAY 18 OF GESTATION. 98.76 108.20 74.22 96.73 96.86 _-__- 19182 P 368. 383. 401. 416. 97.35 19183 P 347. 367. 381. 404. a 19184 P 361. 383. 399. 413. 105.22 19185 P 341. 358. 379. 413. 105.29 19186 P 19187 NP 19188 NP 325. 242. 258. 338. 236. 249. 356. 238. 259. 369. 247. 273. 109.11 _-___---- 19189 P 338. 356. 371. 400. 100.95 19190 P 356. 374. 395. 404. 112.90 19191 P 328. 339. 358. 386. 91.49 19192 P 312. 320. 335. 346. 82.96 19193 P 333. 342. 351. 375. 87.86 19194 P 327. 346. 361. 381, 94.12 19195 P 318. 341. 353. 373. 107.11 19196 P 345. 371. 377. 404. 119.73 19197 P 19198 P 284. 301. 327. 354, FOUND DEAD ON DAY 17 OF GESTATION. 9_8-._99_ _ 19199 P 337. 355. 364. 388. 81.41 19200 P 360. 377. 391. 422. 112.41 .................................................................................................................................... P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Gravid uterine weight not recorded. w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL T O X I C I T Y STUDY O F POTASSIUM PERFLUOROBZPTANE SULFONATE ( P F B S ) I N RATS ( S P O N S O R ' S STUDY NUMBER: T-7485.12) TABLE 17 (PAGE 1 ) : MATERNAL FEED CONSUMPTION VALUES - I N D I V I D U A L DATA -_--_--_________________________________-----.---------------------------- RAT # GROUP I VEHICLE o MG/KG/DAY __--_-____._____________________________------.------------------ PREGNANCY STATUS DAYS 0 - 6 6 - 9 9 - 12 12 - 15 15 - 18 18 - 20 20 - 21 __-__--_________________________________-------------------------------------------------------------------------------------------- 19101 P 137. 61. 63. 69. 79. 52. 24. 19102 P 125. 65. 67. 71. 74. 54. 29. 19103 P 142. 56. 70. 67. 69. 50. 30. 19104 P 138. 70. 69. 73. 78. 52. 24. 19105 P 152. 77. 79. 84. 94. 61. 30. . 19106 P 143. 70. 73. 75. 81. 56. 27. 19107 P 195. 76. 75. 83. 72, 54. 34. 19108 P 120. 61. 67. 62. 68. 49. 27. 19109 P 119. 61. 63. 68. 67. 32. 22. 19110 P 130. 63. 64. 71. 75. 56. 33. 19111 P 138. 64. 65. 65. 68. 54. 27 .a 19112 P 133. 70. 64. 67. 75. 48. 26. 19113 P 136. 70. 67. 69. 76. 51. 29. 19114 P 143. 71. 72. 71. 72. 53. 32. 19115 P 126. 67. 66. 74. 65. 44. 10. 19116 P 126. 69. 62. 71. 86. 61. 28. 19117 P 146. 73. 81. 79. 73. 57. 27. 19118 P 134. 67. 69. 69. 64. 45. 30. 19119 P 121. 61. 61. 70. 74. 52. 24. 19120 P 134. 67. 60. 78. 84. 61. 26. 19121 P 142. 74. 84. 83. 85. 58. 30. 19122 P 136. 65. 66. 80. 88. 63. 28. 19123 P 128. 75. 84. 93. 96. 68. 30. 19124 P 136. 70. 68. 77. 78. 57. 28. 19125 P 128. 63. 58. 70. 82. 51. 24. .................................................................................................................................... P = PREGNANT N P = NOT PREGNANT (VALUES EXCLUDED DAYS = DAYS O F PRESUMED GESTATION ALL WEIGHTS WERE RECORDED I N GRAMS ( G ) . a . Dam delivered on day 21 of gestation. FROM AVERAGES) 4 CA P c PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 17 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - INDIVIDUAL DATA ---____.-__________.____________________------.----------------------- RAT # GROUP I1 LOW DOSAGE 100 MG/KG/DAY ---____.-__________.____________________~..~--.-------------------- PREGNANCY STATUS DAYS 0 - 6 6 - 9 9 - 12 12 - 1 5 15 - 18 18 - 20 20 - 21 ____.____________.______________________----~----------------- 19126 P 134. 59. 68. 82. 84. 58. 28. 19127 P 155. 81. 71. 80. 86. 61. 28. 19128 P 139. 70. 59. 64. 79. 52. 21. 19129 P 127. 71. 73. 81. 88. 58. 31. 19130 P 128. 65. 62. 66. 67. 38. 24. 19131 NP 108. 51. 56. 53. 51. 38. 15. 19132 P 140. 52. 65. 68. 75. 41. 23. 19133 P 128. 65. 68. 73. 77. 51. 30. 19134 P 122. 64. 60. 68. 82. 51. 23. 19135 P 173. 68. 70. 72. 72. 53. 27. 19136 P 152. 70. 67. 76. 78. 50. 17. 19137 P 133. 56. 70. 66. 84. 47. 25. 19138 P 135. 69. 68. 71. 74. 48. 24. 19139 P 131. 63. 70. 73. 66. 44. 20. 19140 P 128. 64. 69. 68. 77. 47. 21. 19141 P 147. 75. 68. 84. 90. 60. 28. 19142 P 130. 63. 69. 69. a 52. 24. 19143 P 134. 73. 67. 70. 76. 49. 18. 19144 P 129. 69. 66. 63. 80. 50. 25. 19145 NP 114. 55. 58. 60. 58. 38. 15. 19146 P 119. 66. 66. 70. 75. 52. 26. 19147 P 151. 76. 74. 76. 72. 52. 28. 19148 NP 127. 62. 61. 53. 50. 36. 19. 19149 P 124. 61. 70. 73, 75. 43. 17. 19150 P 140. 70. 68. 75. 84. 53. 27. __________-_____________________________-------------.~------ P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a . Spilled feed precluded the calculation of this value. 19151 P 19152 NP 132. 65. 71. 78. 81. 56. 25. 121. 58. 51. 47. 48. 37. 16. 19153 P 19154 P 150. 65. 64. 77. 86. 61. 31. 126. 53. 61. 64. 59. 46. 21. 19155 P 146. 73. 73. 77. 78. 52. 25. 19156 P 124. 58. 64. 72. 72. 50. 20. 19157 P 150. 73. 63. 71. 76. 36. 27.b 19158 P 152. 80. 76. 82. 95. 56, 21. 19159 P 131. a 71. 72. 83. 52. 15. 19160 P 147. 61. 71. 80. 71. 53. 22. 19161 P 19162 P 140. 78. 80. 84. 81. 56. 27. 125. 65. 61. 61. 70. 45. 19. 19163 P 126. 64. 61. 58. 60. 50. 23. 19164 P 172. 99. 94. 75. 89. 61. 17. 19165 P 124. 69. 76. 69. 77. 56. 16. 19166 P 136. 69. 73. 72. 80. 54. 27. 19167 P 153. 66. 73. 71. 78. 52. 25. 19168 P 127. 63. 65. 71. 80. 60. 24. 19169 P 154. 82. 82. 82. 86. 53. 28. 19170 P 131. 55. 65. 61. 68. 38. 16, 19171 P 99. 72. 61. 66. 83. 54. 22. 19172 P 19173 P 158. 90. 93. 84. 86. 56. 25. 152. 80. 82. 76. 84. 57. 18. 19174 P 120. 61. 67. 68. 73. 44. 16. 19175 P 132. 71. 63. 76. 80. 52. 24. - _ _ _ _ _ _ _ . _ _ - - - _ _ _ - - _ - - - - - - - - - - - - - . . - . . - . . - . . - . . - . . - . . - . . - . . - . . - . . - . . .-. .-. .-. .-. .-. .-. .- - - - - - - - - - - - - - - - - - - - P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS ( G ) . a . Spilled feed precluded the calculation of t h i s value. b. Dam delivered on day 2 1 of gestation. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 17 (PAGE 4 ) : MATERNAL FEED CONSUMPTION VALUES - INDIVIDUAL DATA -------_-_______..______________________~~------------------------ RAT # GROUP IV HIGH DOSAGE io00 MG/KG/DAY ---_-___________________________________-----.-------------.---------- PREGNANCY STATUS DAYS 0 - 6 6- 9 9 - 12 12 - 15 15 - 18 18 - 20 20 - 21 ------_____r____________________________--.----------------------------------------------------------------------------------------- 19176 P 122. 62. 56. 58. 69. 48. 22. 19177 P 141. 69. 56. 76. 85. 58. 25. 19178 P 117. 65. 52. 63. 62. 47. 20. 19179 P 122. 72. 57. 66. 89. 56. 23. 19180 P 145. 77. 58. 78. 87. 58. 23. 19181 NP 137. 43. 49. 55. FOUND DEAD ON DAY 18 OF GESTATION. 19182 P 152. 76. 77. 77. 86. 53. 16. 19183 P 127. 68. 69. 74. 78. 50. 23. 19184 P 143. 69. 58. 81. 85. 51. 22. 19185 P 145. 64. 60. 77. 70. 46. 29. 19186 P 110. 60. 63. 56. 68. 41. 16. 19187 NP 112. 49. 44. 49. 52. 32. 16. 19188 NP 140. 74. 64. 56. 55. 32. 18. 19189 P 152. 80. 83. 63. 72. 50. 30. 19190 P 140. 71. 70. 66. 79. 53. 14. 19191 P 140. 57. 53. 62. 71. 45. 25. 19192 P 117. 40. 54. 64. 77. 42. 15. 19193 P 133. 59. 71. 68. 71. 39. 23. 19194 P 130. 60. 66. 75. 74. 51. 21. 19195 P 129. 49. 55. 64. 69. 46. 20. 19196 P 137. 70. 73. 65. 63. 39. 21. 19197 P 111. 37. 59. 68. 58. 40. 28. 19198 P 128. 44. 67. 67. FOUND DEAD ON DAY 17 OF GESTATION 19199 P 137. 69. 71. 72. 70. 58. 25. 19200 P 156. 76. 75. 74. 56. 51. 26. _-__-________-__.---____________________------.-------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). R w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 18 (PAGE 1): CAESAREAN-SECTIONINGOBSERVATIONS - INDIVIDUAL DATA -------.--_______._.____________________~~-~-~.-~-----~~~~~~ GROUP I VEHICLE o MG/KG/DAY ----------_.______._____________________-~---~-------------------- _ _ VIABLE ___ FETUSES _.__ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS _ __ __ __ __ __ _ . _ _ _ _ _ _ _ _ - - - _ . __ __ __ __ __ __ _ _ _ _ _ _IM__P-_L-_A-N-T-A_T_IO_N_S_IT_E_S____ C__O_ R_ P_ O_ R_ A_ _ L_ U_ T_ _E_A_ SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT .RIGHT LEFT RIGHT LEFT RAT# M F --------- -____ _ _ _ H_ ORN_ _ _ _ T_ O_ T_ A_ L_ _ _ _ H_ OR_N_ _ _ TOTAL --___ _ _ HORN __ __ - _T- O- T_A_L_ _ _ _ H_ ORN- - - - T_ O_ T_ _A_L _ - -H_ORN_ _ _ _ T_ O_ T_ A_ L_ _ _ _ O_ VAR_ _Y_ _ T_ O_ T_ A_ L_ 19101 12 7 11 8 19 0 0 0 0 0 0 0 0 0 11 8 19 11 8 19 19102 8 7 6 9 15 0 0 0 0 0 0 0 0 0 6 9 15 6 9 15 19103 7 8 7 8 15 0 0 0 0 0 0 0 0 0 7 8 15 7 9 16 19104 5 7 10 2 12 0 0 0 0 0 0 0 0 0 10 2 12 10 4 14 19105 8 7 10 5 15 0 0 0 0 0 0 0 0 0 10 5 15 10 8 18 19106 8 5 9 4 13 0 0 0 1 0 1 0 0 0 10 4 14 10 4 14 19107 6 11 8 9 17 0 0 0 0 0 0 0 0 0 8 9 17 10 12 22 19108 8 6 7 7 14 0 0 0 0 1 1 0 0 0 7 8 15 8 10 18 19109 7 5 4 8 12 0 0 0 0 0 0 0 0 0 4 8 12 6 8 14 19110 8 9 9 8 17 0 0 0 0 0 0 0 0 0 9 8 17 11 9 20 19111 DELIVERED AND SACRIFICED ON DAY 21 OF GESTATION % 19112 11 7 5 13 18 0 0 0 0 0 0 0 0 0 5 13 18 6 13 19 cn\o 19113 7 8 6 9 15 0 0 0 0 0 0 0 0 0 6 9 15 8 9 17 19114 11 5 6 10 16 0 0 0 2 0 2 0 0 0 8 10 18 8 10 18 19115 12 6 12 6 18 0 0 0 0 0 0 0 0 0 12 6 18 13 6 19 19116 4 7 7 4 11 0 0 0 1 1 2 0 0 0 8 5 13 8 7 15 19117 6 6 4 8 12 0 0 0 0 0 0 0 0 0 4 8 12 10 9 19 19118 7 9 10 6 16 0 0 0 0 1 1 0 0 0 10 7 17 11 12 23 19119 6 6 7 5 12 0 0 0 2 0 2 0 0 0 9 5 14 11 6 17 19120 6 8 6 8 14 0 0 0 0 0 0 0 0 0 6 8 14 8 10 18 19121 11 8 9 10 19 0 0 0 0 0 0 0 0 0 9 10 19 12 15 27 19122 4 3 7 0 7 0 0 0 0 0 0 0 0 0 7 0 7 10 12 22 19123 8 8 9 7 16 0 0 0 0 0 0 0 0 0 9 7 16 9 9 18 19124 9 8 8 9 17 0 0 0 0 1 1 0 0 0 8 10 18 8 15 23 19125 10 8 9 9 18 0 0 0 0 0 0 0 0 0 9 9 18 9 12 21 ___________________-____________________------.---------------- M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 18 (PAGE 2 ) : CAESARERN-SECTIONINGOBSERVATIONS - INDIVIDUAL DATA --_--___--______________________________-..----------------------- GROUP I1 LOW DOSAGE loo MG/KG/DAY ------__--______________________________--~----.----------------- VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS _ _ _ _ . _ _ _ _ _ __ _ _ _ _ __ __ __ _ -_ __ __ __ _.__ _ _ _ __ _-- _ --- _ -_ _- __ -__ -_ _ __ _ _ _ _ _ _ _ _ _ _ IM_ P_ _L_A_N_T_A_T_I_O_N_ S_ I_ T_ _ES _ _ C_ O_ R_ P_ O_ R_ A_ _L_U_ T_ E_ A_ _ SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT 'RIGHT LEFT RIGHT LEFT RAT# M F --_---__- _____ HORN TOTAL ____ ____ _____ _ _ _ _HORN TOTAL _-._.- - - _ HORN TOTAL - _ _ _ _ _ _ -- - - - - - - _H_ORN_ _ _ _ T_ O_ T_ A_ _L _ _ _ H_ ORN_ _ _ _ TOTAL ____ - _ _ OVARY -_ _- _ _T_ O_ T_ A_ _L 19126 4 9 7 6 13 0 0 0 0 0 0 0 0 0 7 6 13 7 6 13 19127 7 8 5 10 15 0 0 0 1 1 2 0 0 0 6 11 17 7 11 18 19128 4 9 5 8 13 0 0 0 0 1 1 0 0 0 5 9 14 6 9 15 19129 7 4 6 5 11 0 0 0 0 0 0 0 0 0 6 5 11 8 7 15 19130 7 6 9 4 13 0 0 0 0 0 0 0 0 0 9 4 13 10 8 18 19131 NOT PREGNANT 19132 10 5 9 6 15 0 0 0 1 0 1 0 0 0 10 6 16 12 10 22 19133 5 10 8 7 15 0 0 0 1 0 1 0 0 0 9 7 16 10 7 17 19134 9 7 11 5 16 0 0 0 0 0 0 0 0 0 11 5 16 11 6 17 19135 9 7 11 5 16 0 0 0 0 0 0 0 0 0 11 5 16 11 5 16 19136 5 9 8 6 14 0 0 0 0 0 0 0 0 0 8 6 14 11 9 20 19137 7 10 11 6 17 0 0 0 1 0 1 0 0 0 12 6 18 12 12 24 4 0 19138 7 6 9 4 13 0 0 0 0 0 0 0 0 0 9 4 13 12 9 21 19139 6 2 6 2 8 0 0 0 3 1 4 0 0 0 9 3 12 10 4 14 19140 8 7 8 7 15 0 0 0 1 1 2 0 0 0 9 8 17 10 9 19 19141 6 7 7 6 13 0 0 0 0 0 0 0 0 0 7 6 13 7 6 13 19142 5 8 9 4 13 0 0 0 0 0 0 0 0 0 9 4 13 10 6 16 19143 10 7 8 9 17 0 0 0 3 0 3 0 0 0 11 9 20 11 13 24 19144 10 7 10 7 17 0 0 0 0 1 1 0 0 0 10 8 18 10 11 21 19145 NOT PREGNANT 19146 5 8 9 4 13 0 0. 0 2 0 2 0 0 0 11 4 15 11 4 15 19147 6 10 9 7 16 0 0 0 0 0 0 0 0 0 9 7 16 11 8 19 19148 NOT PREGNANT 19149 8 8 10 6 16 0 0 0 0 0 0 0 0 0 10 6 16 10 9 19 19150 9 8 9 8 17 0 0 0 0 0 0 0 0 0 9 8 17 9 8 17 ___-____-____--___----------------.-----------.--------------------------------- M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE, PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 18 (PAGE 3 ) : CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA ---_-_._________._______________________.----.----------------------- GROUP 111 MIDDLE DOSAGE 3 0 0 MG/KG/DAY ---_--_.__._______._____________________~~~-~.----------~--------- VIABLE FETUSES ________________________ DEAD FETUSES _____ _EA_ _ R_ L_ Y_ _ _ R_ E__ S_ O_ R__ P_ T_ I_.O_ _ N_ _S LATE RESORPTIONS _ - __ _ __ _ __ _ _ _ _ - - IMPLANTATION - _-_- _ - - - _ _ _ SITES CORPORA _ _ - _ ____- _ - - - _ LUTEA _-_-_ _ _ _ _ SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT# --------- M F --__- HORN TOTAL - . - _ _ _ _ _ _ _ _ _ _ _ - _HO_RN_ _ _ _ T_ O_ T_ A_ L_ _ _ _ H_ ORN_ _ _ _ TOTAL --___ HORN ---_ ____ TOTAL ---__ HORN TOTAL ---- -__----_- OVARY TOTAL ___- -_-- --_-_ 19151 5 9 9 5 14 0 0 0 2 0 2 0 0 0 11 5 16 11 7 18 19152 NOT PREGNANT 19153 11 4 5 10 15 0 0 0 1 0 1 0 0 0 6 10 16 6 10 16 19154 5 7 7 5 12 0 0 0 0 0 0 0 0 0 7 5 12 7 5 12 19155 9 10 10 9 19 0 0 0 0 0 0 0 0 0 10 9 19 12 9 21 19156 4 5 5 4 9 0 0 0 2 1 3 0 0 0 7 5 12 8 5 13 19157 DELIVERED AND SACRIFICED ON DAY 2 1 OF GESTATION 19158 4 11 8 7 15 0 0 0 0 0 0 0 0 0 8 7 15 8 7 15 19159 6 9 7 8 15 0 0 0 0 0 0 0 0 0 7 8 15 9 8 17 19160 9 8 12 5 17 0 0 0 0 1 1 0 0 0 12 6 18 12 6 18 2 19161 4 9 10 3 13 0 0 0 0 1 1 0 0 0 10 4 14 10 4 14 % 19162 7 6 10 3 13 0 0 0 0 1 1 0 0 0 10 4 14 10 5 15 2 19163 7 6 4 9 13 0 0 0 1 0 1 0 0 0 5 9 14 5 10 15 19164 6 10 14 2 16 0 0 0 0 0 0 0 0 0 14 2 16 14 2 16 19165 12 3 5 10 15 0 0 0 0 0 0 0 0 0 5 10 15 5 10 15 19166 10 7 9 8 17 0 0 0 0 0 0 0 0 0 9 8 17 9 9 18 19167 6 10 7 9 16 0 0 0 0 0 0 0 0 0 7 9 16 7 9 16 19168 7 8 6 9 15 0 0 0 1 0 1 0 0 0 7 9 16 7 9 16 19169 10 7 10 7 17 0 0 0 0 1 1 0 0 0 10 8 18 13 8 21 19170 5 8 6 7 13 0 0 0 0 1 1 0 0 0 6 8 14 6 9 15 19171 7 6 12 1 13 0 0 0 1 0 1 0 0 0 13 1 14 13 1 14 19172 10 5 7 8 15 0 0 0 0 0 0 0 0 0 7 8 15 7 8 15 19173 6 6 9 3 12 0 0 0 0 1 1 0 0 0 9 4 13 11 7 18 19174 4 8 4 8 12 0 0 0 0 0 0 0 0 0 4 8 12 4 8 12 19175 7 8 6 9 15 0 0 0 0 0 0 0 0 0 6 9 15 6 9 15 .................................................................................................................................... M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 18 (PAGE 4): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA __--___-________________________________-----.-------------------------- GROUP IV HIGH DOSAGE 1000 MG/KG/DAY ________________________________________-------------------------------------------------------------------------------------------- VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA _ _ _ _ _ _ _ _ _ ___ _ _ __ _ _.__ _ __ _ __ -___ _- _ _ -_- -_ _ __ - - -_ - - __ ---_ - - - _ - - - - - - _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - _ _ - - - _ _ SEX RAT# M F _ _ - - _ _ _ _ __ _ _ _ _ RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT . RIGHT LEFT RIGHT LEFT _ _ _ H_ ORN_ _ _ _ TOTAL _____ _ _ _ H_ ORN_ _ _ _ TOTAL HORN TOTAL - _ _ _ __ _ _ - _ _ - - _ _ - - - - _ _HO_RN_ _ _ _ TOTAL ---__ HORN TOTAL - _ _ - _ _ - -- _ _ - - OVARY TOTAL _--_ ---_ _____ 19176 7 8 11 4 15 0 0 0 0 0 0 0 0 0 11 4 15 12 6 18 19177 9 8 7 10 17 0 0 0 0 1 1 0 0 0 7 11 18 7 11 18 19178 7 4 6 5 11 0 0 0 3 0 3 0 0 0 9 5 14 9 I 16 19179 7 6 8 5 13 0 0 0 0 0 0 0 0 0 8 5 13 8 6 14 19180 4 10 8 6 14 0 0 0 1 1 2 0 0 0 9 7 16 9 7 16 19181 FOUND DEAD ON DAY 18 OF GESTATION 19182 10 5 7 8 15 0 0 0 1 0 1 0 0 0 8 8 16 9 11 20 19183 7 7 7 7 14 0 0 0 1 1 2 0 0 0 8 8 16 10 9 19 19184 4 11 11 4 15 0 0 0 0 0 0 0 0 0 11 4 15 11 5 16 19185 10 5 8 7 15 0 0 0 0 0 0 0 0 0 8 I 15 8 7 15 % 19186 8 8 8 8 16 0 0 0 1 1 2 0 0 0 9 9 18 9 10 19 4 19187 NOT PREGNANT td 19188 NOT PREGNANT 19189 9 6 9 6 15 0 0 0 0 3 3 0 0 0 9 9 18 15 9 24 19190 9 7 5 11 16 0 0 0 0 0 0 0 0 0 5 11 16 5 11 16 19191 6 8 7 7 14 0 0 0 0 0 0 0 0 0 7 7 . 14 11 9 20 19192 8 5 5 8 13 0 0 0 1 0 1 0 0 0 6 8 14 8 9 17 19193 9 5 8 6 14 0 0 0 0 0 0 0 0 0 8 6 14 10 8 18 19194 8 5 4 9 13 0 0 0 0 0 0 0 0 0 4 9 13 4 9 13 19195 9 7 8 8 16 0 0 0 0 0 0 0 0 0 8 8 16 10 8 18 19196 6 12 10 8 18 0 0 0 0 0 0 0 0 0 10 8 18 10 8 18 19197 8 8 7 9 16 0 0 0 1 0 1 0 0 0 8 9 17 9 9 18 19198 FOUND DEAD ON DAY 17 OF GESTATION 19199 4 9 11 2 13 0 0 0 0 2 2 0 0 0 11 4 15 12 5 17 19200 7 11 12 6 18 0 0 0 0 0 0 0 0 0 12 6 18 13 6 19 ________________.-______________________----------------------- M = MALE F = FEMALE PLACENTAE APPEARED NORMAL UNLESS NOTED OTHERWISE. PROTOCOL 418-023: ORAL (GAVAGE)DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 19 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA 19101 19102 19103 19104 19105 19106 19107 19108 19109 19110 19111 19112 19113 19114 19115 19116 19117 19118 19119 19120 19121 19122 19123 19124 19125 12 7 19 5.73 5.32 5.58 19 8 7 15 5.50 5.22 5.37 15 7 8 15 5.34 5.16 5.24 15 5 7 12 5.46 5.13 5.26 12 8 7 15 5.65 5.31 5.49 15 8 5 13 6.29 5.88 6.14 14 6 11 17 5.41 5.16 5.25 17 8 6 14 6.12 5.89 6.02 15 7 5 12 5.12 4.95 5.05 12 8 9 17 5.50 5.51 5.50 17 DELIVERED AND SACRIFICED ON DAY 21 OF GESTATION 11 7 18 5.07 4.61 4.89 18 7 8 15 5.32 5.00 5.15 15 11 5 16 5.37 5.30 5.35 18 12 6 18 4.96 4.57 4.83 18 4 7 11 5.94 5.62 5.74 13 6 6 12 5.64 5.42 5.53 12 7 9 16 4.96 4.64 4.78 17 6 6 12 5.39 5.18 5.29 14 6 8 14 5.50 5.03 5.23 14 11 8 19 5.45 5.04 5.28 19 4 3 7 6.09 5.59 5.88 7 8 8 16 5.76 5.71 5.74 16 9 8 17 4.73 4.30 4.53 18 10 8 18 5.10 4.86 4.99 18 0 0.0 0 0.0 0 0.0 0 0.0 0 0.0 1 7.1 0 0.0 1 6.7 0 0.0 0 0.0 0 0.0 0 0.0 2 11.1 0 0.0 2 15.4 0 0.0 1 5.9 2 14.3 0 0 .o 0 0.0 0 0.0 0 0.0 1 5.6 0 0.0 b w N PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 1 9 (PAGE 2): LITTER OBSERVATIONS (CRESAREAN-DELIVERED FETUSES) - INDIVIDUAL DATA 19126 4 19127 7 19128 4 19129 7 19130 7 19131 NOT 19132 10 19133 5 19134 9 19135 9 19136 5 19137 7 19138 7 19139 6 19140 8 19141 6 19142 5 19143 10 19144 10 19145 NOT 19146 5 19147 6 '19148 NOT 19149 8 19150 9 ___--___---__-. 9 8 9 4 6 PREGNANT 5 10 7 7 9 10 6 2 7 7 8 7 7 PREGNANT 8 10 PREGNANT 8 8 13 6.01 5.50 5.65 13 0 15 5.60 5.32 5.46 17 2 13 5.71 5.34 5.45 14 1 11 5.98 5.52 5.81 11 0 13 5.46 5.18 5.33 13 0 15 5.34 5.31 5.33 16 1 15 6.02 5.53 5.69 16 1 16 5.16 5.32 5.57 16 0 16 5.24 4.76 5.03 16 0 14 6.10 5.74 5.87 14 0 17 5.26 4.78 4.98 18 1 13 4.81 4.47 4.65 13 0 8 5.56 5.38 5.52 12 4 15 5.40 5.25 5.33 17 2 13 5.54 5.05 5.28 13 0 13 6.13 5.75 5.89 13 0 17 5.32 5.14 5.25 20 3 17 5.20 4.97 5.10 18 1 13 5.36 5.09 5.19 15 2 16 5.18 5.02 5.08 16 0 16 5.24 4.89 5.06 0 17 5.58 5.24 5.42 0 ____-_____________------------------- 0.0 11.8 7.1 0.0 0.0 6.2 6.2 0.0 0.0 0.0 5.6 0.0 33.3 11.8 0.0 0.0 15.0 5.6 13.3 0.0 PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR`SSTUDY NUMBER: T-7485.12) TABLE 19 (PAGE 3): LITTER OBSERVATIONS (CAESRREAN-DELIVEREDFETUSES) - INDIVIDUAL DATA RAT # MALE FEMALE TOTAL MALE - - - - - - - - - - _ _ _ _ _ _ _ _ _ _ _ _ ______ _ _ _ _ _ _ _ _ _ _ _ _ _ _ - - - - - - - - - - - - - - - - - 19151 5 9 14 5.08 16 19152 NOT PREGNANT 19153 11 4 15 5.30 5.09 5.25 16 19154 5 7 12 5.60 5.37 5.46 12 19155 9 10 19 4.77 4.52 4.64 19 19156 4 5 9 6.09 5.99 6.03 12 19157 DELIVERED AND SACRIFICED ON DAY 21 OF GESTATION 19158 4 11 15 4.98 4.78 4.83 15 19159 6 9 15 5.32 4.78 4.99 15 5cb 19160 9 8 17 5.15 4.94 5.05 18 19161 4 9 13 5.62 5.31 5.40 14 19162 7 6 13 5.18 4.79 5.00 14 19163 7 6 13 5.30 5.30 5.30 14 19164 6 10 16 4.95 4.70 4.79 16 19165 12 3 15 5.13 5.02 5.11 15 19166 10 7 17 5.21 4.85 5.06 17 19167 19168 19169 6 10 16 5.47 5.12 5.25 16 7 a 15 5.63 5.40 5.51 16 10 7 17 5.24 4.94 5.12 18 19170 5 8 13 5.11 5.07 5.08 14 19171 7 6 13 5.48 5.37 5.43 14 19172 10 5 15 5.28 5.05 5.20 15 19173 19174 19175 6 6 12 5.30 4.83 5.06 13 4 a 12 5.01 4.75 4.84 12 7 a 15 5.67 5.23 5.44 15 2 12.5 1 6.2 0 0.0 0 0.0 3 25.0 0 0.0 0 0.0 1 5.6 1 7.1 1 7.1 1 7.1 0 0.0 0 0.0 0 0.0 0 0.0 1 6.2 1 5.6 1 7.1 1 7.1 0 0.0 1 7.7 0 ' 0.0 0 0.0 PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 19 (PAGE 4): LITTER OBSERVATIONS (CAESAREAN-DELIVEREDFETUSES) - INDIVIDUAL DATA 19176 19177 19178 19179 19180 19181 19182 19183 19184 19185 19186 19187 19188 19189 19190 19191 19192 19193 19194 19195 19196 19197 19198 19199 19200 7 8 15 5.19 4.77 4.97 15 9 8 17 4.85 4.42 4.65 18 7 4 11 4.98 4.61 4.84 14 7 6 13 5.13 5.12 5.13 13 4 10 14 5.17 4.84 4.93 16 FOUND DEAD ON DAY 18 OF GESTATION 10 5 15 4.72 4.70 4.71 16 7 7 14 5.01 4.56 4.79 16 4 11 15 5.23 4.95 5.02 15 10 5 15 5.42 5.06 5.30 15 8 8 16 5.34 5.04 5.20 18 NOT PREGNANT NOT PREGNANT 9 6 15 5.06 4.88 4.99 18 9 7 16 5.10 4.64 4.90 16 6 8 14 4.80 4.82 4.82 14 8 5 13 4.80 4.76 4.78 14 9 5 14 4.69 4.38 4.58 14 8 5 13 5.27 4.88 5.12 13 9 7 16 4.79 4.53 4.67 16 6 12 18 4.97 4.70 4.79 18 8 8 16 4.90 4.64 4.77 17 FOUND DEAD ON DAY 17 OF GESTATION 4 9 13 4.95 4.56 7 11 18 4.66 4.56 0 0.0 1 5.6 3 21.4 0 0.0 2 12.5 1 6.2 2 12.5 0 0.0 0 0.0 2 11.1 3 0 0 1 0 0 0 0 1 2 0 _--- 16.7 0.0 0.0 7.1 0.0 0.0 0.0 0.0 5.9 13.3 0.0 P c ? 0 N w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL T O X I C I T Y STUDY O F POTASSIUM PERFLUOROBUTANE SULFONATE ( P F B S ) I N RATS ( S P O N S O R ' S STUDY NUMBER: T-7485.12) TABLE 2 0 (PAGE 1 ) : FETAL S E X , V I T A L STATUS AND BODY WEIGHT - INDIVIDUAL DATA RAT # C L s 19101 11/ 8 19102 6/ 9 19103 7/ 9 19104 10/ 4 19105 10/ 8 19106 10/ 4 19107 10/12 19108 8/10 19109 6/ 8 19110 11/ 9 19111 MA MA 5.95 5.61 MA MA 5.93 5.33 MA F A 5.39 5.31 FA FA 4.84 4.89 MA FA 5.78 5.27 MAMA 6.17 6.49 FA MA 4.79 5.13 FA MA 6.22 5.85 F A MA 5.05 4.91 MA F A 5.82 5.23 DELIVERED F A MA MA 5.43 5.69 5.79 FA FA FA 5.19 5.27 5.40 MA FA MA 5.32 5.53 5.46 F A FA MA 5.30 5.50 5.43 MA FA MA 5.41 5.21 5.37 M A M A MA 6.61 6.39 6.27 F A MA FA 5.58 5.59 5.22 MA MA MA 6.47 6.14 6.03 MA MA / MA 5.46 5.17 4.95 MA FA FA 5.57 5.46 5.66 AND S A C R I F I C E D FA MA F A F A MA F A / M A 5.50 5.85 5.22 5.33 6 . 0 0 5.27 6.00 FA / FA FA FA M A M A M A 5.40 4.95 5.23 5.12 5.27 5.49 5.39 MA M A / F A MA MA FA F A 5.52 5.44 4.86 4.85 5.41 4.91 5.04 MA MA F A F A M A / F A MA 5.33 5.57 4.90 5.07 5.47 5.40 5.48 MA F A F A MA FA / FA MA 5.56 5.07 5.50 5.74 5.48 5.33 6.07 E F A MA F A FA / MA MA 5.76 6.36 5.62 6.06 6.20 5.86 MA MA F A / F A F A MA F A 5.76 5.13 5.18 4.70 5.18 5.49 5.48 FA M A / F A E F A MA F A 5.99 6.23 5.64 5.94 5.95 5.93 F A F A F A MA MA MA F A 4.84 4.98 5.28 4.78 5.43 5.13 4.60 MA MA F A FA / F A MA MA 5.11 5.32 5.19 5.49 6.01 5.29 5.38 ON DAY 2 1 O F GESTATION FA 5.17 MA 5.52 FA 5.49 MA 5.49 MA 5.43 FA 5.09 MA 5.44 MA 5.64 FA 5.06 FA 5.31 FA 5.95 FA 5.18 FA 5.61 MA 5.51 FA 6.02 FA 5.15 MA 6.43 MA 5.75 FA 5.16 MA 5.84 F A MA F A 5.41 5.51 5.53 MA MA 6.10 5.28 F A MA 5.13 5.35 F A MA 5.62 5.97 FA 5.36 MA 5.53 19112 19113 6/13 8/ 9 MA 4.96 FA 5.47 MA MA 5.29 4.87 F A MA 4.96 5.91 MA 5.09 MA 5.83 M A / F A FA 5.30 4.12 4.11 MA FA / FA 5.23 5.15 5.08 F A MA 4.80 5.14 MA F A 4.83 5.21 MA 5.16 FA 4.43 MA 4.95 FA 4.52 FA 4.81 MA 5.15 MA 4.72 MA 5.20 FA 4.89 FA 5.15 FA 4.55 MA 5.12 MA 4.94 FA 5.02 MA 5.34 M = MALE F = FEMALE A = A L I V E E = EARLY RESORPTION " / " DENOTES P O S I T I O N OF CERVIX CLS = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED I N GRAMS (GI. P c 90 8 w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T - 7 4 8 5 . 1 2 ) TABLE 20 (PAGE 2): FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA RAT # CLS 19114 6/10 FA MA MA MA E MA E M A / M A MA MA FA MA FA FA FA M A M A 5.44 5.62 5.59 5.37 6.00 4.69 5.35 4.86 5.14 5.00 5.52 5.43 5.36 5.26 5.45 5.53 19115 13/ 6 MA MA MA MA MA MA MA FA FA FA FA F A / M A MA MA MA MA FA 19116 e/ 7 5.43 4.89 5.06 4.86 4.52 5.32 4.95 4.34 4.43 4.49 4.54 4.80 4.96 5.09 4.85 4.96 4.62 4.82 FA MA E FA FA MA MA FA / FA FA E MA FA 5.67 5.70 5.78 5.46 6.05 5.92 5.55 5.76 5.46 6.09 5.68 19117 10/ 9 FA FA MA F A / M A MA FA MA MA FA MA FA 5.30 5.63 5.16 5.64 5.97 5.54 5.39 5.64 5.56 5.14 5.94 5.41 19118 11/12 MA FA FA MA MA MA FA FA FA FA / MA MA E FA FA MA FA 5.15 4.85 5.12 5.44 4.81 4.83 4.57 4.88 4.53 3.86 4.12 4.94 4.95 4.35 5.40 4.64 19119 11/ 6 E FA E MA FA FA MA MA M A / F A FA MA MA FA 19120 8/10 5.07 5.16 5.11 4.92 5.42 5.45 5.66 5.38 5.27 5.31 5.34 5.35 FA MA MA FA FA MA / FA FA FA MA MA FA FA MA 5.34 5.41 5.70 4.85 4.04 5.78 5.55 5.13 4.86 5.51 5.34 5.10 5.34 5.28 19121 12/15 MA FA FA MA FA MA FA MA FA / MA FA FA MA MA MA FA M A M A M A 5.46 4.80 5.05 5.77 5.22 5.39 5.56 5.83 5.35 5.21 4.65 4.84 5.36 5.32 5.33 4.83 5.29 5.62 5.38 19122 10/12 FA MA FA FA MA MA M A / 5.37 6.23 5.84 5.57 6.03 6.25 5.86 19123 9/ 9 FA MA FA FA MA MA FA MA MA / FA FA MA MA MA FA FA 5.67 5.85 5.79 5.56 5.97 5.57 5.64 5.32 6.10 5.52 5.84 5.57 5.80 5.94 5.80 5.86 19124 8/15 MA MA FA MA FA MA MA M A / F A E FA MA MA MA FA FA FA FA 4.92 5 .OO 5.58 4.45 4.49 4.71 4.03 4.71 2.65 4.81 5.30 4.34 5.09 4.48 4.31 4.23 3.87 19125 9/12 MA MA MA FA FA MA MA FA MA / FA MA FA FA FA MA MA FA MA 5.08 5.06 4.92 4.86 4.95 5.13 5.18 4.67 5.38 4.80 4.83 4.94 4.84 5.04 5.06 4.80 4.82 5.51 .................................................................................................................................... M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION 'I/" DENOTES POSITION OF CERVIX CLS = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (GI. PROTOCOL 4 1 8 - 0 2 3 : ORAL (GAVAGE) DEVELOPMENTAL T O X I C I T Y STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE ( P F B S ) I N RATS ( S P O N S O R ' S STUDY NUMBER: T - 7 4 8 5 . 1 2 ) TABLE 2 0 (PAGE 3 ) : FETAL S E X , V I T A L STATUS AND BODY WEIGHT - I N D I V I D U A L DATA __________._____________________________-..-------------------- GROUP I1 LOW DOSAGE 1 0 0 MG/KG/DAY _________________.______________________-.--------------------- FETUS# 1 2 3 4 5 6 7 8 9 1 0 11 1 2 1 3 1 4 1 5 1 6 1 7 18 1 9 2 0 ________________________________________-.---.--------------------- RAT # 19126 CLs 7/ 6 19127 7/11 19128 6/ 9 19129 8/ 7 1 9 1 3 0 10/ 8 19131 MA FA FA 5.95 5.69 5.69 FA FA E 5.24 5.51 FA MA MA 5.37 5.74 5.66 MA MA FA 6.22 6.38 5.76 MA FA FA 5.66 5.42 5.19 NOT PREGNANT MA 6.15 FA 5.81 FA 5.19 MA 5.98 MA 5.42 MA MA F A / F A 6.30 5.64 5.33 5.57 MA MA / F A E 5.57 5.56 4.29 FA / MA F A F A 5.55 5.50 5.64 4.97 FA F A / MA MA 5.75 5.05 5.44 5.92 MA MA F A MA 5.33 5.66 5.10 5.50 FA FA 5.57 5.37 MA FA 5.61 5.19 MA FA 5.94 4.87 MA MA 5.94 5.96 MA / FA 5.28 5.05 FA 5.59 MA 5.55 FA 5.41 FA 5.52 FA 5.04 FA 5.48 MA 5.45 E MA 5.35 FA 5.18 MA 5.47 FA 5.46 FA 5.64 FA 5.59 FA 5.28 FA 5.27 FA 5.65 MA 6.02 1 9 1 3 2 1 2 / 1 0 MA MA FA E MA. MA F A F A F A MA / M A MA FA M A M A M A 5.38 5.33 5.27 5.20 5.57 5.54 4.88 5.60 5.54 5.49 5.02 5.25 5.12 5.14 5.56 1 9 1 3 3 10/ 7 F A MA E FA FA F A F A F A F A / FA M A M A M A F A F A MA 5.50 5.73 5.96 5.23 5.44 5.62 5.30 5.72 5.33 5.73 6.56 6.10 5.51 5.69 5.97 1 9 1 3 4 11/ 6 MA FA MA FA FA M A M A MA M A M A FA / FA F A MA F A MA 1 9 1 3 5 11/ 5 5.40 5.10 5.75 4.99 5.44 5.69 5.66 5.95 6.27 5.59 5.54 5.67 5.03 5.96 5.45 5.57 F A MA MA FA FA F A MA F A M A M A MA / FA F A M A M A M A 3.00 5.08 5.28 5.09 5.05 5.14 5.61 4.99 5.38 4.99 4.11 4 . 8 1 5.24 5.80 5.40 5.55 1 9 1 3 6 11/ 9 MA MA FA MA FA F A F A MA I F A FA FA MA FA 6 . 1 1 6.12 5.25'6.03 5.75 5.76.5.87 6.11 6.00 5.54 5.84 6.15''k:lO FA 5.60 19137 12/12 FA 4.70 E MA FA FA MA F A F A MA FA MA F A / M A F A MA F A MA F A 5.21 3.63 5.14 5.51 4.76 4.95 4.81 4.63 5.04 5.21 5.61 4 . 4 1 5.30 5.12 5.38 5.11 1 9 1 3 8 1 2 / 9 F A FA FA MA FA MA FA MA M A / M A FA MA MA 4.49 4.12 4.60 5.02 4.77 4.95 4.23 5.03 3.04 4.98 4.61 5.16 5.49 --- M = MALE F = FEMALE A = A L I V E E = EARLY RESORPTION ' I / " DENOTES P O S I T I O N O F CERVIX CLS = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED I N GRAMS ( G ) . P c 90 0 wh) PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL T O X I C I T Y STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE ( P F B S ) I N RATS ( S P O N S O R ' S STUDY NUMBER: T-7485.12) TABLE 20 (PAGE 4 ) : FETAL S E X , V I T A L STATUS AND BODY WEIGHT - INDIVIDUAL DATA RAT # CLS 19139 10/ 4 19140 10/ 9 19141 7/ 6 19142 10/ 6 19143 11/13 19144 10/11 19145 MA F A E 5.27 5.11 MA MA FA 5.10 5.71 5.34 FA FA MA 5.19 5.20 5.59 FA FA MA 5.82 5.85 6.21 MAEE 5.11 MAMAMA 5.33 5.36 5.21 NOT PREGNANT MAE 5.70 FA MA 5.32 5.96 FA FA 3.80 5.08 FA FA 5.61 5.75 MA MA 5.73 5.53 MAMA 5.14 4.76 MA 5.67 MA 5 .72 FA 5.22 MA 6.28 MA 5.47 FA 4 .80 MAMA 5.41 5.63 E FA 5.29 FA / MA 5.51 5.11 F A MA 5.32 5.93 MA MA 4.94 5.00 FA FA 4.90 4.70 E/FA E MA 5.64 5.69 MA / FA FA MA E 5.22 5.05 5.27 4.65 MAFAMAMAMA 5.66 5.34 5.54 5.45 5.91 FA / MA FA MA F A 6.13 5.99 6.02 6.23 5.47 MA E FA / FA FA 5.68 5.38 5.36 5.18 MA F A / F A FA MA 5.40 4.90 4.92 5.19 5.13 MA 5.62 FA 5.18 MA 5.26 FA 5.11 MA 5.58 MA 5.37 MA 5.25 FA 4.75 MA 5.05 FA 5.35 MA 4.91 E FA 5.15 FA 5.36 FA 5.02 MA 5.29 03 0 19146 11/ 4 FA E FA MA FA MA F A FA MA MA E / FA FA FA MA 4.74 5.14 4.84 5.30 5.58 5.19 4.91 5.59 5.48 4.98 5.18 5.25 5.32 19147 11/ 8 F A F A F A MA FA F A MA F A MA / FA MA MA FA MA F A FA 5.44 5.01 4.46 5.15 4.90 5.33 5.36 4.78 5.09 4.73 4.98 5.12 5.41 5.39 4.96 5.19 19148 NOT PREGNANT 19149 10/ 9 FA MA F A FA FA MA MA F A MA M A / M A FA FA MA F A MA 4.58 5.25 4.16 5.16 4.96 5.31 5.13 5.02 5.22 4.89 5.56 5.44 4.78 5.35 5.00 5.22 19150 9/ 8 F A MA FA MA MA MA F A MA FA / FA MA MA F A FA MA MA FA 5.32 5.67 5.21 5.51 5.17 5.46 4.85 5.88 5.20 4.80 5.99 5.36 5.74 5.26 5.50 5.64 5.52 ________________________________________-------------------------------------------------------------------------------------------- M = MALE F = FEMALE A = A L I V E E = EARLY RESORPTION " / " DENOTES P O S I T I O N O F CERVIX C L S = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED I N GRAMS ( G ) . PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 2 0 (PAGE 5 ) : FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA 19153 6/10 19154 7/ 5 19155 12/ 9 19156 E/ 5 19157 E MA 5.66 FA FA 5.55 4.99 MA FA 4.79 4.71 FA FA 5.80 5.90 DELIVERED FA MA MA 5.06 5.01 5.39 FA FA FA 5.74 5.56 5.37 FA MA MA 4.71 4.84 4.72 MA E E 6.04 AND SACRIFICED F A / M A MA MA MA MA 5.12 5.07 5.46 4.92 5.68 5.27 FA MA / MA FA MA MA 5.28 5.64 5.51 5.10 5.26 5.67 MA FA FA MA FA / FA 4.78 4.49 4.45 4.47 4.16 4.63 M A F A / M A M A FA FA 6.18 6.29 5.94 6.20 5.58 6.36 ON DAY 21 OF GESTATION MA 4.84 MA 5.91 MA 4.85 E FA 5.03 MA 4.74 MA 5.53 FA 4.76 MA 5.53 MA 4.94 FA 5.14 MA 4.84 FA 4.73 FA 4.15 FA 4.44 0C-0L 19158 E/ 7 FA FA FA FA FA FA FA FA / MA FA MA FA MA FA MA 4.68 4.34 4.69 4.69 4.78 4.96 5.08 5.13 4.96 4.66 4.98 4.55 4.83 5.01 5.13 19159 9/ 8 FA FA FA FA MA MA FA / MA FA FA MA MA FA FA MA 4.61 5.00 4.71 4.60 5.37 5.43 4.83 5.22 4.66 4.60 4.98 5.49 5.02 4.95 5.40 19160 12/ 6 FA MA MA MA MA MA FA MA FA FA MA FA / FA MA E FA MA FA 4.79 5.41 5.03 4.80 5.70 5.06 5.08 4.72 4 . E O 4.61 4.96 5.22 4.78 5.42 19161 10/ 4 FA MA FA FA FA FA MA FA MA FA / MA FA E FA 5.22 5.27 4.99 4.73 5.11 5.04 5.47 5.05 5.03 5.86 5.41 5.55 5.68 5.95 5.58 5.77 19162 10/ 5 MA FA FA MA FA MA FA MA FA F A / E MA MA MA 4.98 4.93 4.81 5.28 4.49 4.97 4.65 5.31 5.24 4.61 5.45 4.92 5.33 19163 5/10 FA MA MA E F A / M A FA FA MA MA FA MA MA FA 5.69 5.35 5.33 _----__________-- 5.37 5.41 5.25 5.17 5.20 5.30 5.06 5.46 5.08 5.27 M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION " / " DENOTES POSITION OF CERVIX CLS = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS ( G ) . PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 20 (PAGE 6 ) : FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA _____-__________________________________-------------------~------------------------------------------------------------------------ GROUP I11 MIDDLE DOSAGE 300 MG/KG/DAY ___________________.____________________-.---.-------------~- FETUS# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 ----_----__.____________________________.~---~--------~~~~---- RAT # CLs 19164 14/ 2 FA MA FA FA FA MA FA FA MA MA FA MA FA FA / MA FA 4.19 4.96 4.35 4.51 4.50 4.86 4 . 9 7 5.00 5.19 4.86 4.64 4.74 4.64 5.11 5.07 5.07 19165 5/10 MA FA MA MA MA / MA MA MA MA MA FA MA MA MA FA 5.82 5.05 5.11 5.42 5.28 4.86 5.09 4.97 4.97 5.21 4.88 5.09 4.84 4.86 5.14 19166 9/ 9 FA FA FA MA MA MA FA FA FA / MA MA FA MA MA MA MA MA 4.70 4.88 4.95 5.16 5.32 5.36 4.80 5.11 4.62 5.39 4.96 4.91 4.94 5.02 5.39 5.12 5.45 19167 7/ 9 FA MA MA MA FA FA FA / FA FA FA MA FA FA FA MA MA 5.57 5.52 5.06 5.72 5.14 5.25 5.02 5.09 5.16 4.91 5.49 5.20 4.79 5.03 5.60 5.42 19168 7/ 9 MA FA MA E MA FA FA / FA FA MA MA MA FA FA MA FA 5.44 5.18 5.73 5.81 5.31 5.52 5.44 5.39 5.34 5.89 5.51 5.22 5.48 5.70 5.70 19169 13/ 8 MA FA MA MA MA FA FA FA MA MA / FA MA FA MA E MA MA FA 4.86 4.82 5.03 5.35 5.40 5.10 4.94 4.90 5.16 5.51 4.89 5.26 4.81 5.17 19170 6 / 9 MA FA FA FA MA FA / FA MA FA FA E FA MA MA 5.31 5.39 5.16 4.96 5.51 5.08 5.21 5.14 5.22 5.01 4.92 4.86 4.75 4.92 5.25 5.28 00 N 19171 13/ 1 MA FA MA MA MA MA FA FA E FA FA FA MA / MA 5.72 5.42 5.18 5.66 5.32 5.65 5.36 5.65 5.33 5.06 5.40 5.26 5.59 19172 7/ 8 MA MA MA MA FA FA FA / MA MA MA FA MA MA FA MA 5.13 5.19 5.36 4.94 4.87 5.19 5.03 5.14 5.27 5.53 4.99 5.29 5.46 5.18 5.49 19173 11/ 7 FA FA MA FA FA MA FA MA MA / FA MA E MA 4.72 4.62 5.12 4.83 5.00 5.02 4.85 5.41 5.64 4.94 5.13 5.49 19174 4/ 8 FA FA MA MA / FA FA MA FA MA FA FA FA 4.91 4.93 4.84 5.07 4.64 4.76 5.08 5.02 5.06 4.33 4.76 4.62 19175 6 / 9 FA FA MA MA MA MA / MA FA FA MA FA FA MA FA FA 4.92 5.48 5.42 5.76 5.75 5.64 5.73 5.04 5.28 5.66 5.31 5.40 5.75 5.27 5.13 _________----___________________________-------------------------------------------------------------------------------------------- M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION 'I/" DENOTES POSITION OF CERVIX CLS = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G). P soC-L 0 w N PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL T O X I C I T Y STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE ( P F B S ) I N RATS ( S P O N S O R ' S STUDY NUMBER: T-7485.12) TABLE 2 0 (PAGE 7 ) : FETAL S E X , V I T A L STATUS AND BODY WEIGHT - I N D I V I D U A L DATA RAT # CLS 19176 12/ 6 19177 7/11 19178 9/ 7 19179 E/ 6 19180 9/ 7 19181 FA FA M A MA MA FA F A MA FA FA 4.39 4.90 5.12 5.09 4.89 4.69 4.42 4.98 4.77 4.75 FA MA MA FA FA FA MA / F A MA MA 4.48 4 .86 5.01 4.43 4.33 4.50 5.10 4.37 4 .82 4.87 MA MA FA E MA E FA E MA / MA 4.79 5.11 4.86 4.88 4.82 5.21 4.89 FA MA MA FA MA FA MA F A / MA MA 4.99 5.43 5.32 5.39 5.39 4.96 4.21 5.10 4.86 5.48 FA FA E FA F A F A F A F A MA / FA 3.43 5.06 4.92 5.19 4.85 5.14 5.19 5.32 4.74 FOUND DEAD ON DAY 18 O F GESTATION FA / MA 4.78 5.61 MA F A 4.68 4.31 FA FA 4.48 4.29 FA FA 5.28 5.03 MA E 4.85 MA 5.58 MA 4.60 MA 4.99 MA 5.24 FA 4.80 MA 5.08 E MA 4.98 MA 5.12 FA 5.46 FA 4.64 MA 5.38 MA 4.90 FA 5.03 FA 4.26 MA 4.85 19182 9/11 M A MA F A MA E MA MA M A / M A MA FA F A MA MA F A F A 4.75 4.89 4.51 4.86 4 .85 4.64 4.85 4.69 4.60 4.82 4.39 4.66 4.44 4.85 4.91 0w 0 19183 10/ 9 MA E FA M A M A M A MA FA / FA E MA F A F A MA F A F A 5.27 4.86 5.13 4.91 4.95 5.16 4.42 4.67 5.01 4.08 4.55 4.67 4.53 4.81 19184 11/ 5 MA FA MA MA FA F A F A F A FA MA FA / F A F A F A F A 5.47 4.65 5.10 5.12 4.99 4.99 5.00 5.06 4.89 5.22 5.05 4.69 4.95 4.99 5.19 19185 E/ 7 F A FA FA M A M A M A MA M A / M A FA MA MA F A M A M A 4.88 4.84 5.06 5.47 5.46 5.08 5.28 5.31 5 . 5 0 4.97 5.36 5.41 5.55 5.76 5.56 19186 9/10 F A MA MA FA F A F A MA E M A / F A MA MA MA F A F A F A EMA 4.95 5.36 5.03 5.14 4.89 4.54 5.41 5.09 5.67 5.41 5.75 5.69 5.45 4.99 4.73 5.02 19187 NOT PREGNANT 19188 NOT PREGNANT . M = MALE F = FEMALE A = A L I V E E = EARLY RESORPTION L = LATE RESORPTION C L s = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED I N GRAMS ( G ) ' I / " DENOTES POSITION OF CERVIX 6 N w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 20 (PAGE 8 ) : FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA ----------______________________________----~~--~--~~~---------------------------------------------------------------------------~-- GROUP IV HIGH DOSAGE 1000 MG/KG/DAY ------_--_______________________________-------------------------------------------------------------------------------------------- FETUS# 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 ----___--_______________________________-------------------------------------------------------------------------------------------- RAT # CLs 19189 151 9 FA FA MA MA M A FA FA MA MA 1 E MA MA MA FA MA E 4.88 4.72 5.24 4.91 4.57 4.44 4.80 4.69 5.17 5.50 5.13 5.14 4.98 5.17 19190 5/11 MA MA MA FA MA / FA FA MA FA MA FA FA FA MA MA MA E FA 5.46 4.76 5.29 5.19 4.87 5.39 4.68 4.91 4.84 3.69 5.11 4.83 4.83 4.71 5.09 5.13 5.11 19191 11/ 9 FA FA FA FA FA MA MA / FA MA MA FA MA MA FA 4.97 4.87 5.09 4.48 4.79 4.55 4.85 4.86 5.06 5.33 5.15 4.46 4.58 4.38 19192 E/ 9 FA E MA MA MA MA / FA MA FA FA MA FA MA MA 5.00 5.13 4.69 4.85 4.99 4.49 4.90 4.91 4.87 4.56 4.51 4.60 4.69 19193 10/ 8 MA FA MA MA FA FA MA MA / MA FA FA MA MA MA 4 .81 4.75 4.95 4.88 4.44 3.77 4.51 4.62 4.37 4.45 4.48 4.79 4.56 4.69 19194 4 / 9 MA MA FA FA / FA FA MA MA MA MA MA FA MA 5.28 5.56 5.02 4.92 4.98 4.92 5.33 5.00 5.34 5.19 4.91 4.55 5.56 19195 10/ 8 FA FA FA MA MA FA MA MA / MA FA MA MA FA FA MA MA 3.79 4.68 4.45 4.50 4.91 4.56 4.79 5.15 4.88 4.52 4.41 4 .EO 4.63 5.08 5.01 4.63 00 P 19196 10/ 8 M A FA MA MA FA FA FA MA MA FA / FA FA FA FA FA MA FA FA 4 .88 4.56 4 .SO 4.99 4 .85 5.02 4.76 5.01 5.15 5.33 4.70 4.55 4.23 4 .SO 4.42 5.31 4.42 5.11 19197 9/ 9 MA FA FA FA MA FA MA E / MA MA MA MA FA FA FA FA MA 4.90 4.25 4.88 4.67 5.00 4.46 4.88 4.89 4.83 4.60 4.78 4.80 4.70 4.72 4.62 5.34 19198 FOUND DEAD ON DAY 17 OF GESTATION 19199 12/ 5 FA FA MA MA FA MA MA FA FA FA FA / E E FA FA 4.57 4.32 4.77 4.76 4.61 5.60 4.68 4.60 4.39 4.74 4.26 5.04 4.56 19200 13/ 6 FA MA MA FA FA FA MA MA FA FA MA FA / MA FA FA FA FA MA 4.59 4.36 4.22 4.43 4.64 4.80 4.47 4.92 4.44 4.00 4.98 4.76 5.02 4.53 4.74 4.34 4.88 4.64 .................................................................................................................................... M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION " / " DENOTES POSITION OF CERVIX CLS = CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS ( G ) . 19101 O ( 0.0) 0/19 0/10 19102 19103 19104 % w 19105 w 19106 03 VI 19107 19108 19109 O ( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) 1( 8 . 3 ) 0/15 0/15 0/12 0/15 0/13 0/17 0/14 0/12 FETUS 2 VESSELS: UMBILICAL ARTERY DESCENDED TO THE LEFT OF THE URINARY BLADDER O/ 8 o/ 8 O/ 6 o/ 8 o/ 7 o/ 9 o/ 7 O/ 6 19110 19111 O ( 0.0) 0/17 o/ 8 DELIVERED AND SACRIFICED ON DAY 21 OF GESTATION a a/ 9 19112 O( 0.0) 0/18 o/ 9 19113 O ( 0.0) 0/15 o/ 7 19114 O ( 0.0) 0/16 O/ 8 o / 19115 O ( 0.0) 0/18 9 I o / i3 19116 O ( 0.0) 0/11 5 w _ _ _ _ _ _ _ _ _ - - _ _ _ - - - . _ _ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED a . Dam 19111 delivered 2 pups and had 13 fetuses and 1 early resorption in utero. examination all pups and fetuses appeared normal. At gross external, s o f t tissue and skeletal PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'S STUDY NUMBER: T-7485.12) TABLE 21 (PAGE 2): FETAL ALTERATIONS - INDIVIDUAL DATA 19118 19119 19120 1( 6.2) 0/16 O ( 0.0) 1( 7.1) 0/12 0/14 l/ 8 FETUS 10 EYES: ABSENT EYE, left 19121 19122 19123 O ( 0.0) O ( 0.0) 1( 6.2) 0/19 a/ 7 0/16 O/ 8 19124 O ( 0.0) 0/17 19125 O ( 0.0) 0/18 o/ 9 N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED O/ 6 1/ 7 FETUS 5 THORACIC VERTEBRAE : CENTRUM, BIFID, 11th; ARCHES, FUSED, left 13th and 14th; HEMIVERTEBRA, left 14th, arch and centrum with attached rib; CENTRUM, UNILATERAL OSSIFICATION, left 14th; CENTRUM, NOT OSSIFIED, 13th 1/ 8 FETUS 11 THORACIC VERTEBRAE: CENTRUM, BIFID, 10th o/ 9 PROTOCOL 418-023: ORAL (GAVAGE)DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 21 (PAGE 3) : FETAL ALTERATIONS - INDIVIDUAL DATA 00 4 19131 19132 NOT PREGNANT O ( 0.0) 0/15 1/ 7 FETUS 9 THORACIC VERTEBRAE: CENTRUM, BIFID, 13th 19133 O ( 0.0) 0/15 19134 O ( 0.0) 0/16 19135 O ( 0.0) 0/16 19136 O( 0.0) 0/14 19137 O ( 0.0) 0/17 19138 2( 15.4) 0/13 1/ 6 FETUS 2 VESSELS: UMBILICAL ARTERY DESCENDED TO THE LEFT OF THE URINARY BLADDER ................................................................................................ N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED FETUS 9 THORACIC VERTEBRAE: d CENTRUM, BIFID, 4th; cn RIBS: FUSED, right 4th and 5th, proximally ______--_________________ P c 00 b E PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 21 (PAGE 4) : FETAL ALTERATIONS - INDIVIDUAL DATA 19139 2 1 25.0) o/ 8 o/ 4 2/ 4 FETUS 7 PELVIS: PUBIS, INCOMPLETELY OSSIFIED, right FETUS 10 THORACIC VERTEBRAE: CENTRUM, BIFID, 11th % 19140 1( 6 . 7 ) 0/15 19141 19142 19143 O( 0.0) O( 0 . 0 ) 1( 5.9) 0/13 0/13 0/17 O/ 6 1/ 8 FETUS 19 EYES: FOLDED RETINA, right 1/ 8 o/ 7 o/ 7 o/ 9 FETUS 12 STERNAL CENTRA: FUSED, 1st and 2nd; ASYMMETRIC, 1st - 3rd 19144 2( 11.8) 0/17 o/ 8 2/ 9 FETUS 1 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, left FETUS 11 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, left 19145 NOT PREGNANT _____________---------------------------------.---------------------------- N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 21 (PAGE 5): FETAL ALTERATIONS - INDIVIDUAL DATA --____-_________________________________-----.-------------------------- GROUP I1 LOW DOSAGE loo MG/KG/DAY ----__.--_______________________________-.------------------------ SPECIMENS GROSS EXTERNAL EXAMINATION SOFT TISSUE EXAMINATION SKELETAL EXAMINATION WITH ANY RAT ALTERATIONS . NUMBER N(%) N/N DESCRIPTION N/N DESCRIPTION N/N DES,CRIPTION _-____---_______________________________-.---------------------- 19146 2 ( 15.4) 0/13 O/ 6 2/ 7 FETUS 8 THORACIC VERTEBRAE : CENTRUM, BIFID, 12th FETUS 13 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, left (3 19147 1( 6.2) 0/16 1/ 8 FETUS 10 VESSELS: UMBILICAL ARTERY DESCENDED TO THE LEFT OF THE URINARY BLADDER O/ 8 19148 NOT PREGNANT 19149 O ( 0.0) 0/16 O/ 8 O/ 8 1 . 9150 ..... . . . . . .O.(. . .0...0.) . . . . . .0./.1.7. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .O./. .8 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .o./ . .9. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED P c w PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 21 (PAGE 6) : FETAL ALTERATIONS - INDIVIDUAL DATA 19152 NOT PREGNANT 19153 O ( 0.0) 0/15 19154 O ( 0.0) 0/12 19155 O ( 0.0) 0/19 19156 19157 19158 19159 O ( 0.0) o/ 9 DELIVERED AND SACRIFICED ON DAY 21 OF GESTATION a O ( 0.0) 0/15 o/ 7 O ( 0.0) 0/15 o/ 7 19160 19161 O ( 0.0) O ( 0.0) 0/17 0/13 O/ 8 O/ 6 o/ 7 19162 19163 19164 19165 19166 O ( 0.0) O ( 0.0) O ( 0.0) O ( 0.0) 1( 5.9) 0/13 0/13 0/16 0/15 0/17 o/ 7 o/ 7 o/ 8 % o / 8 Q c 1/ 9 FETUS 9 CA THORACIC VERTEBRAE: P c CENTRUM, BIFID, 12th 19167 0/16 o / w O ( 0.0) 8 ______-__---------------------------------.--.-------------------------------- N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED a. Dam 19157 delivered 1 pup and had 16 fetuses in utero. A t gross external, soft tissue and skeletal examination the pup and all fetuses appeared normal. PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBWANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 2 1 (PAGE 7) : FETAL ALTERATIONS - INDIVIDUAL DATA 19169 1( 5.9) 0/17 19170 19171 19172 O( 0.0) O ( 0.0) 1( 6.7) 0/13 0/13 0/15 O/ 6 O/ 6 o/ 7 1/ 9 o/ 7 o/ 7 1/ 8 FETUS 18 THORACIC VERTEBRAE: CENTRUM, BIFID, llth FETUS 3 THORACIC VERTEBRAE: CENTRUM, BIFID, llth 4 CA P Y PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 2 1 (PAGE 8): FETAL ALTERATIONS - INDIVIDUAL DATA 19178 19179 19180 734 19181 \o til 19182 O ( 0.0) 0/11 o/ 5 O ( 0.0) 0/13 O/ 6 O( 0.0) 0/14 o/ 7 FOUND DEAD ON DAY 18 OF GESTATION (NOT PREGNANT) 1( 6.7) 0/15 o/ 7 O/ 6 o/ 7 o/ 7 1/ 8 FETUS 3 CERVICAL VERTEBRAE: CERVICAL RIB PRESENT AT 7TH CERVICAL VERTEBRA, right 19183 1( 7.1) 0/14 o/ 7 1/ 7 FETUS 4 THORACIC VERTEBRAE: CENTRUM, BIFID, 11th 19184 O ( 0.0) 0/15 o/ 7 o/ 8 19185 1( 6.7) 0/15 1/ 7 FETUS 8 o/ 8 VESSELS: INNOMINATE ARTERY ABSENT % 19186 O ( 0.0) 0/16 o/ 8 o/ 8 Q c 1 9 1 8 7 NOT PREGNANT v1 s" . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . P N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED c 0 wtil PROTOCOL 418-023: ORAL (GAVAGE) DEVELOPMENTAL TOXICITY STUDY OF POTASSIUM PERFLUOROBUTANE SULFONATE (PFBS) IN RATS (SPONSOR'SSTUDY NUMBER: T-7485.12) TABLE 21 (PAGE 9 ) : FETAL ALTERATIONS - INDIVIDUAL DATA 19189 O ( 0.0) 0/15 o/ 7 O/ 8 19190 19191 19192 19193 O( 0.0) O ( 0.0) O ( 0.0) O( 0.0) 0/16 0/14 0113 0/14 O/ 8 o/ 7 O/ 6 o/ 7 O/ 8 o/ 7 o/ 7 o/ 7 19194 O ( 0.0) 0/13 O/ 6 o/ 7 19195 O ( 0.0) 0/16 O/ 8 O/ 8 19196 O ( 0.0) 0/18 o/ 9 o/ 9 19197 O ( 0.0) 0/16 O/ 8 O/ 8 19198 FOUND DEAD ON DAY 17 OF GESTATION a 19199 O ( 0.0) 0/13 O/ 6 o/ 7 19200 O ( 0.0) 0/18 o/ 9 o/ 9 ________________________________________-------------------------------------------------------------------------------------------- N/N = NUMBER OF SPECIMENS WITH ALTERATIONS/NUMBER OF SPECIMENS EXAMINED a. Dam 19198 was found dead on day 17 of gestation and had 15 fetuses and one early resorption present in utero. A t gross external examination all fetuses appeared normal for early developmental age. Fetuses could not be further examined due to early developmental age. APPENDIX C PROTOCOL Page 94 ARGUS 418-023 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Honham. PA 19044 Telephone: (215) 443-8710 Telefax: (215) 443-8587 PROTOCOL 418423 SPONSOR'S STUDY NUMBER: T-7485.12 STUDY TITLE: PURPOSE: TESTING FACILITY: STUDY DIRECTOR: SPONSOR: STUDY MONITOR: Oral (Gavage) Developmental Toxicity Study of Potassium Perfluorobutane Sulfonate (PFBS) in Rats The purpose of this study is to evaluate the developmental toxicity (embryo-fetal toxicity and teratogenic potential) of Potassium Perfluorobutane Sulfonate (PFBS) administered orally via gavage to Crl:CD@(SD)IGSBR VAF/PIu& presumed pregnant female rats. Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Telephone: (215) 443-8710 Telefax: (215) 443-8587 Raymond G. York, Ph.D., DABT Associate Director of Research raym0nd.yot-k 8 primedica.com 3M Corporate Toxicology 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 Paul H. Lieder, Ph.D., DABT 3M Corporate Toxicology 3M Medical Department Telephone: (651) 737-2678 Telefax: (651) 733-1773 Page 95 ARGUS 418-023 REGU LATORY CITATIONS: Protocol 418-023 Paas 2 U.S. Environmental ProtectionAgency (1998). Health Effects Test Guidelines; Prenatal DevelopmentalToxicity Study. Office of Prevention, Pesticides and Toxic Substances (OPPTS) 870.3700, August, 1998, U.S. Environmental Protection Agency (1997). Toxic Substances Control Act (TSCA) Test Guidelines; Final Rule. Prenatal Developmental Toxicity, 799.9370 (crossreferenced to OPPTS 870.3700). Federal Register, August 15, 1997. Organization for Economic Cooperation and Development (1981). OECD Guidelines for Testing of Chemicals. Section 4, No. 414: Teratogenicity, adopted 12 May 1981. Japanese Ministry of Agriculture, Forestry and Fisheries (1985). Guidance on Toxicology Study Data for Application of Agricultural Chemical Registration. 59 NohSan No. 4200. U.S. Environmental Protection Agency. Toxic Substances Control Act (TSCA); Good Laboratory Practice Standards; Final Rule. 40 CFR Part 792. US. Environmental Protection Agency. Federal Insecticide, Fungicide and Rodenticide Act (FIFRA); Good Laboratory Practice Standards; Final Rule. 40 CFR Part 160. Organization for Economic Cooperation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. Japanese Ministry of Agriculture, Forestry and Fisheries (1984). Good Laboratory Practice Standards. 59 NohSan No. 3850. REGULATORY COMPLIANCE: This study will be conducted in compliance with the Good Laboratory Practice (GLP) regulations cited above. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. The Testing Facility's Quality Assurance Unit (QAU) will audit the protocol, the raw data and the report, and will inspect critical phases of those portions of the study conducted at the Testing Facility in accordance with the Standard Operating Procedures of Argus Research Laboratories, Inc. Should any portion of the study be conducted by a subcontractor or by the Sponsor, the QAU for that facility will conduct critical phase inspections and audit respective results and reports for that study portion according to the SOPSof that facility. Such critical phase inspection reports and report audits will be submitted by that facility to the Study Director. The dates of the inspections and report submissions will be incorporated into Page 96 ARGUS 418-023 Protocol 418-023 Page 3 a QAU Statement generated by that facility and provided to the Testing Facility for inclusion in the final report. The final report will include a compliance statement signed by the Study Director that the report accurately reflects the raw data obtained during the performance of the study and that all applicable GLP regulations were followed in the conduct of the study. Should significant deviations from GLP regulations occur, each will be described in detail, together with how the deviation might affect the quality or integrity of the study. SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. TEST SUBSTANCE AND VEHICLE: ldentification: Test Substance: Potassium Perfluorobutane Sulfonate (PFBSor T-7485). Lot number 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 purity of the test substance. Vehicle: Aqueous 0.1 o/o carboxymethylcellulose (CMC) (medium viscosity) prepared using reverse osmosis membrane processed deionized water (R.O. deionized water). Lot number will be documented in the raw data. Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this protocol will be conducted, Safetv Precautions: Gloves, dust-misVHEPA-filtered mask, appropriate eye protection and uniform/lab coat to be worn during formulation preparation and dosage. In addition, Tyvek sleeves and a half-face respirator will be worn and preparation will be conducted in a chemical fume hood when using the bulk test substance. The Material Safety Data Sheet is attached to the protocol (ATTACHMENT 2). Page 97 ARGUS 418-023 Storaqe: Protocol 418-023 Page 4 Bulk Test Substance: Bulk Vehicle Components: Prepared Test Substance and Vehicle Formulations: Room temperature. Room temperature. 2C to 0C All test substance shipments to the Testing Facilrty should be addressed to the attention of Julian Gulbinski, Manager of Formulations, at the previously cited address and telephone number. Shipments should include information concerning storage conditions and shipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment. FORMULATlON: Frequencv of Preparation: Formulations (suspensions) will be prepared approximately every 10 days at the Testing Facility. Detailed preparation procedures will be attached to this protocol (ATTACHMENT3). Adiustment for Purity: The test substance will be considered 100% pure for the purpose of dosage calculations. Testinq Facilitv Reserve Samples: The Testing Facility will reserve a 1 g sample of each lot of bulk test substance and vehicle 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 deemed necessary during the course of the study. Page 98 ARGUS 4 18-023 Bulk Test Substance Samplinq: Protocol 418-023 Page 5 A sample (1 g) of the test substance will be taken on the last day of treatment and sent (ambient conditions) to the Sponsor for analysis. This sample will be sent to: James D. Johnson Southern Research Institute 2000 Ninth Avenue South Birmingham, Alabama 35205 Telephone: (205) 581-2715 (205) 581-2599 Telefax: ,(205) 322-7473 (lab phone) The recipient will be notified in advance of sample shipment. Analvses of Prepared Formulations: Homoqeneity: Homogeneity analyses of the preparedformulationswill not be conducted. Information is on file with the Sponsor to document the homogeneity of the test substance in the prepared vehicle over the range of concentrationsto be used in this study. This information will be providedto the Study Directorfor inclusion in the final report. Concentration: Concentration of the prepared formulations will be verified during the course of this study. Duplicate samples (2 mL each) will b e taken from the first and last preparation on the day prepared. One sample of each set will be shipped for analysis; the remaining samples will be retained at the Testing Facility as backup samples. Backup samples will be stored under the previously cited conditions and discarded at the Testing Facility upon the request of the Sponsor. Stability: Stability data for prepared formulations bracketingthe range of concentrations in this study are on file with the Sponsor and will not be determined during the conduct of this study. This information will be provided to the Study Director for inclusion in the final report. Page 99 ARGUS 418-023 Shippincr Instructions: PtotOCOl418-023 Page 6 Samples to be analyzed will be shipped (on cold packs) to James D. Johnson, at the previously cited address. 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: Nelda Marecki, Ph.D. 3M SMMG EHS&R 3M Center Building 236-16-10 St. Paul, Minnesota 55144-1000 TEST SYSTEM: SpeciedStrainand Reason for Selection: The Crl:CDB(SD)IGS BR VAF/PIu& rat was selected as the Test System because: 1) it is one mammalian species accepted and widely used throughout industry for nonclinical studies of developmental toxicity (embryo-fetal toxicity/teratogenicity); 2) this strain has been demonstrated to be sensitive to developmental toxins; and 3) historical data and experience exist at the Testing Facility('-3). Number: Initial population acclimated: Population selected for study: 140 virgin female rats. 100 mated female rats (25 per dosage group). Bodv Weiqht and Acle: Female rats will be ordered to have body weights of 200 g to 225 g each at receipt, at which time they will be expected to be at least 60 days of age. Actual body weights recorded the day after receipt will be documented in the raw data, and the weight range will be included in the final report. - Sex: Female rats will be given the test substance. Male rats of the same source and strain will be used only as breeders and are not considered part of the Test System. Page 100 ARGUS 418-023 Source: Protocol 418-023 Page 7 Charles River Laboratories, Inc. The rats will be shipped in filtered cartons by air freight and/or truck from Charles River Laboratories, Inc., to the Testing Facility. Identification: Rats are permanently identified using MoneRO self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats are given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats are assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study on the basis of day 0 of presumed gestation body weights. ANIMAL HUSBANDRY: All cage sizes and housing conditions are in compliance with the Guide for the Care and Us8 of Laboratory Animald4). Housing: The rats will be individually housed in stainless steel, wire-bottomed cages except during the cohabitation period. During cohabitation, each pair of rats will be housed in the male rat's cage. No nesting materialswill be supplied because the female rats will be sacrificed before parturition is expected. Room Air, Temperature and Humidity: The animal room is independentlysupplied with at least ten changes per hour of 100% fresh air that has been passed through 99.97sb HEPA filters. Room temperature will be maintainedat 64F to 79F (18C to 26C)and monitoredconstantly. Room humidity will also be monitored constantlyand maintainedat 30% to 70%. Liqht: An automatically-controlled12-hour light:12-hour dark fluorescent light cycle will be maintained. Each dark period will begin at 1900 hours EST. - Diet: Rats will be given Certified Rodent Diet@#5002 (PMI Nutrition International)available a d libitum from individual feeders. Page 101 ARGUS 418-023 - Water: Protocol 418-023 Page 8 Water will be available ad libitum from individual bottles attached to the cages or from an automatic watering access system. All water will be from a local source and passed through a reverse osmosis membrane before use. Chlorine will be added to the processed water as a bacteriostat: processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. Contaminants: Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the certified diet or in the drinking water at levels that would interfere with the results of this study. Therefore, no analyses other than those routinely performed by the feed supplier or those mentioned in this protocol will be conducted. RANDOMIZATION AND COHABITATION: Upon arrival, male and female rats will be assigned to individual housing on the basis of computer-generated random units. After acclimation, virgin female rats will be cohabited with breeder male rats, one male rat per female rat. The cohabitation period will consist of a maximum of five days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ will be considered to be at day 0 of presumed gestation and assigned to individual housing. Healthy mated female rats will be assigned to dosage groups based on computergenerated (weight-ordered) randomization procedures. ADMINISTRAT10N: Route and Reason for Choice: The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered; and 2) it is one possible route of human exposure. Method and Frequency: Female rats will be given the test substance once daily on days 6 through 20 of presumed gestation. Dosages will be adjusted daily for body weight changes and given at approximately the same time each day. Rationale for Dosaqe Selection: Dosages were selected on the basis of a dosage-range study (Argus Research Laboratories, Inc., Protocol 418-023P). Page 102 ARGUS 4 18-023 Protocol 41 8-023 Page 9 The highest dosage level selected should induce some overt developmentalandor maternal mortality. The intermediate dosage levels should produce minimal observed toxic effects. The lowest dosage level should not produce any evidence of either maternalor developmentaltoxicity. Dosaae Levels, Concentrations and Volumes: IV 25 loo0 100 10 The test substance wrll be considered 100% pure for the purposeof dosage calculabons. 8418-023-D(Day.Month.Year) TESTS, ANALYSES AND MEASUREMENTS: Viability: All Periods: At least twice daily. Clinical Observations andor General Appearance: Acclimation Period: Weekly. Predosage Period: Day 0 of presumed gestation. Dosage Period: Daily before dosage. Postdosage observations will be recorded approximately 60 10 minutes after administration. Day of Sacrifice: Once. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director andor Study Monitor. Bodv Weiqhts: Acclimation Period: Weekly. Predosage Period: Day 0 of presumed gestation. Dosage Period: Daily. Day of Sacrifice: Prior to sacrifice. Page 103 ARGUS 418-023 Feed Consumption Values (recorded and tabulated): Protocol 418-023 Page 10 Predosage Period: Day 0 of presumed gestation. Dosage Period: Days 6, 9, 12, 15, 18 and 20 of presumed gestation. Day of Sacrifice: Feed left recorded. Feed consumption values may be recorded more frequently if it is necessary to replenish the feed. These intervals will not be tabulated. Matinq Performance: Mating will be evaluated daily during the cohabitation period and confirmed by observation of spermatozoa in a smear of the vaginal contents and/or a copulatory plug observed in situ. Caesarean-Sectioninq Observations: Rats will be Caesarean-sectioned on day 21 of presumed gestation. To minimize bias, Caesarean-sectioning and subsequent fetal observations will be conducted without knowledge of dosage group. The gravid uterus will be excised and weighed. The fetuses will be removed from the uterus and placed in individual containers. The rats will be examined for number and distribution of: Corpora Lutea. Implantation Sites. Live and Dead Fetuses. (A live fetus is defined as one that responds to stimuli; a dead fetus is defined as a term fetus that does not respond to stimuli and that is not markedly autolyzed; dead fetuses demonstrating marked to extreme autolysis are considered to be late resorptions.) Early and Late Resorptions. (A conceptus is defined as a late resorption if it is grossly evident that organogenesis has occurred; if this is not the case, the conceptus is defined as an early resorption.) Page 104 ARGUS 418-023 Fetal Obsenrations: Protocol 418-023 Page 11 To minimize bias, fetal observations will be conducted without knowledge of dosage group. Gross External Alterations and Sex: Fetuses will be examined for sex and for gross external alterations. Late resorptions and dead fetuses also will be examined for sex and for gross external alterations to the extent possible but such observations will not be included in either data summarization or statistical analyses. Body Weiqhts and Identification: The body weight of each fetus will be recorded. Only body weights of live fetuses will be used to determine litter fetal body weight averages. Fetuses will be tagged with identification noting study number, litter number, uterine distribution and fixative. Soft Tissue Examination: Approximately one-half of the fetuses in each litter will be examined for soft tissue alterations by using a variation of the microdissection technique of Staples('). These fetuses will be fixed in Bouin's solution and the heads will subsequently be examined by free-hand sectioning: head sections will be stored in alcohol. The decapitated carcasses will not be retained. Skeletal Examination: The remainingfetuses (approximately one-half of the fetuses in each litter) will be examined for skeletal alterations (bone and cartilage) after staining with alizarin red S(6). The fetuses will be initially fixed in alcohol; skeletal preparations will be retained in glycerin with thymol added as a preservative. Representative photographs of fetal gross, soft tissue and skeletal alterations will be taken. METHOD OF SACRIFICE: Rats will be sacrificed by carbon dioxide asphyxiation. Live fetuses will be sacrificed by an intraperitoneal injection of sodium pentobarbital. NECROPSY: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation. Unless specifically cited below, all other tissues will be discarded. Page 105 ARGUS 418-023 Scheduled Sacrifice: Protocol 418-023 Page 12 On day 21 of presumed gestation, female rats will be Caesarean-sectioned, and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. The gravid uterus will be excised and weighed. Uteri of apparently nonpregnant rats will be examined while being pressed between glass plates to confirm the absence of implantation sites. Rats Found Dead or Moribund: Rats that die or are sacrificed because of moribund condition, abortion or premature delivery will be examined for the cause of death or moribund condition on the day the observation is made. The rats will be examined for gross lesions. Pregnancy status and uterine contents of female rats will be recorded. Gravid uterine weights will not be recorded if precluded by autolysis. Aborted fetuses, delivered pups and/or concepti in uteri will be examined to the extent possible, using the methods described for term fetuses. Uteri of apparently nonpregnant rats will be examined while being pressed between glass plates to confirm the absence of implantation sites. Page 106 ARGUS 418-023 Protocol 41 8-023 Page 13 PROPOSED STATISTICAL METHODS(7"3' Averages and percentageswill be calculated. Litter values will be used where appropriate. Additional procedures andor analyses may be performed, if appropriate. T w e of TesP I. Parametric II. Nonparametricb r l A. Bartlett'sTest' I Significant at ps.0.001 Not Significant A. Kruskal-WallisTest r - l(~75%ties) Significant at ~50.05 Not Significant Significant at ps0.05 Not Significant 6. Fisher's Exact Test (>75% ties) Dunnett's Test Ill. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilitiesare reportedas either ps0.05 or ps0.01. b. Proportion data are not included in this category. c. Test for homogeneity of variance. Page 107 ARGUS 418-023 DATA ACQUISITION, VERIFICATION AND STORAGE: Protocol 418-023 Page 14 Data generated during the course of this study will be recorded either by hand or using the Primedica Argus Automated Data Collection and Management System and the Vivarium Temperature and Relative Humidity Monitoring System. All data will be tabulated, summarized and/or statistically analyzed using the Primedica Argus Automated Data Collection and Management System, the Vivarium Temperature and Relative Humidity Monitoring System, Microsoft Excel [part of Microsoft Office 97 (version SR-2)] and/or The SAS System (version 6.12). Records will be reviewed by the Study Director and/or appropriate management personnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft final report, after which time the Sponsor will be contacted to determine the disposition of these materials. RECORDS TO BE MAINTAINED: Protocol and Amendments. Test Substance, Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (if prescribed by Staff Veterinarian). Generat Comments. Clinical Observations and/or General Appearance. Blood Sample Collection, Processing and Shipment (if required). Body Weights. Feed ConsumptionValues. Caesarean-Sectioning and Fetal Observations. Gross Necropsy observations. Organ Weights (if required). Photographs (if required). Study Maintenance (room and environmental records). Feed and Water Analyses. Packing and/or Shipment Lists. Page 108 ARGUS 418-023 KEY PERSONNEL: Protocol 418-023 Page 15 Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Director of Research: Alan M. Hoberman, Ph.D., DABT Associate Director of Research and Study Director: Raymond G. York, Ph.D., DABT Director of Laboratory Operations: John F. Bamett, B.S. Manager of Animal Operations: Dena C. Lebo, V.M.D. Chairperson, InstitutionalAnimal Care and Use Committee: Theresa Woodard, D.V.M. Director of Operations and Compliance: Barbara J. Patterson, B.A. Director of Study Management: Valerie A. Sharper, M.S. Consultant, Veterinary Pathology: W. Ray Brown, D.V.M., Ph.D., ACVP FINAL REPORT: The Study Director will provide periodic updates of study progressto the Sponsor. Draft summary tables of unaudited 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 followingconsultationwith the Sponsor. i h e report will include the following: Summary and Conclusion. Experimental Design and Method. Evaluation of Test Results. Appendices: Figures, Summary and IndividualTables Summarizing the Above Data, Protocol and Associated Amendments and Deviations, Study Directots GLP Compliance Statement, Reports of Supporting Data (if appropriate) and QAU Statement. The Sponsor will receive one copy of the draft report and two copies of the final report. INSTITUTIONAL ANIMAL CARE AND USE COMMIlTEE STATEMENT: The procedures described in this protocol have been reviewed by the Testing Facility's InstitutionalAnimal Care and Use Committee. All procedures described in this protocol that involve study animals will be conducted in a manner to avoid or minimize discomfort, distress or pain to the animals. The Sponsor's signature below documents the fact that informationconcerningthe 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 meetingthe stated purposes of the study. Page 109 ARGUS 418-023 REFERENCES: PtotOCOl418-023 Page 16 1. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical InformationService, U S .Department of Commerce, Springfield, VA 22161. 2. Christian, M.S. (1984). Reproductive toxiclty and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 3. Lang, P.L. (I 988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charfes River Crl:CLXDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 4. Institute of Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Staples, R.E. (1974). Detection of visceral alterations in mammalian fetuses. Teratology 9(3):A37-38. 6. Modification of method of Staples, R.E. and Schnell, V.L. (1964). Refinement in rapid clearing technique in the KOH-alizarin red S method for fetal bone. Stain Technol. 39:61-63. 7. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneity of the binomial distribution. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 240-241. 8. Sokal, R.R. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, W.H. Freeman and Co.,S a n Francisco, pp. 370-371. 9. Snedecor, G.W. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, 6th Edition, Iowa State University Press, Ames, pp. 258-275. 10. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50~096-1129. 11. Sokal, R.R. and Rohlf, F.J. (1969). Kruskal-WallisTest. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 12. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241-252. 13. Siegel, S. (1956). Nonparametric Statistics for the Behavioral Sciences, McGraw-Hill, New York, pp. 96-104. Page 110 PROTOCOL APPROVAL: FOR THE TESTING FACILITY ARGUS 418-023 Protocol 418-023 Page 17 Study Director Theresa Woodard, D.V.M. Chairperson, InstitutionalAnimal Care and Use Committee FOR THE SPONSOR --L-kEedQ---/----------------- Date Paul H. Lieder, Ph.D., DABT Study Monitor and Sponsor's Representative Page 111 ARGUS 4 18-023 AlTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE Page 112 AITACHMENT 1 ARGUS 418-023 Protocol 418-023 Page 1 of 2 STUDY SCHEMATIC DEVELOPMENTAL TOXICITY STUDYa Female Rats ' Start of Dosage Cohabitation I Day 6 of Presumed Gestation End of Dosage I CaesareanSectioningb Day 20 of Presumed Gestation Day 21 of Presumed Gestation = Dosage Period. a. For additional details see "Tests, Analyses and Measurementsnsection of the protocol. b. Fetal evaluations (all fetuses - external examinations , one-half the fetuses in each litter - soft tissue or skeletal examinations). Page 113 . AlTACHMENT 1 ARGUS 418-023 Protocol 418-023 Page 2 of 2 SCHEDULEa 13 FEB 01 18 FEB 01 PM - 23 FEB 01 AM 19 FEB 01 23 FEB 01 25 FEB 01 '- 15 MAR 01 12 MAR 01 - 16 MAR 01 06 JUN 01 Animals Arrive - Acclimation Begins. Cohabitation Period. First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. Dosage Period (Days 6 through 20 of presumed gestation). Caesarean-Sectioning Period (Day 21 of presumed gestation). Draft Final Report. ....................... a. The study initiation date is the date the Study Director signs the protocol. Page 114 ARGUS 418-023 AlTACHMENT 2 MATERIAL SAFETY DATA SHEET Page 115 ARGUS 418-023 CUTERIM SAfElY DATA (~xpari.ontW 3n Sn Centor st. P8ul, nillnomtr 561u-100a 1-800-W-357) Of (8S1) 131-6501 (24 hOUrS) capyrlght, 2000, nlnnorot. M h g 8nd nurufrcturing company. All rights nwrvmd. Coprina andlor dbwiLolding of t h i s inforration t o r thr plrpo8@Ot prop.rly utiUzing 3)1 prpQlcts e is 8Uouad provFkd that: 1) tlm i n f o r r r r t b l Is bl fau t h no changes unless prior agn.rant i 8 obtainad f r a Sl, and 2) neithrr thr copy nor th. original Is mrold o r othrnrite distributed With tha intantion o t earning a p r o f i t thereon. DIVISION: 3R spH;]IALTy HAl'ERuLs rwTERuL: L-7038 DEVUO-AL WERUL ISSUED: Dacnber 0 8 , 2000 SUPERSEDES: October 11, 2000 -: 04-4134-2 -9(1 -2 The caponWItS Of thh product 8rC in cmpli8nca u i t h the c h m i o r l n o t i f i c a t i o n nquirsllmtr of fieA. A l l 8ppliCable chemical ingrrdientS in t h i s mterfrl 8- U s t s d on the Europe811 Invmntory o f Existing C h c l i c i l Substanca8 (U)(Ecs),o r i r e exempt polymers nhose monomers a r t l i s t e d on EINECS. VBOAIKLMINGPRPESOSINUTR:E..:............................... NlA UIA VWAAPOpRoRmAnwmI RA:TE.:............................. MIA WIA SSOPELCLIllFBIICUTOyRIANVrM rV:E.R..:.. .................... 5 % c.. 0.54 blatepl PERCENT MLATILE: .............. Bulk MIA DmSity 7-8 lb/~8l pH: ............................ 5.5 .7 . S VISCOSITY:. .................... (SI Aqueous) MIA MELTING POINT:. ................ 285 C APPEARANCE AND OWR: S o l i d , uhhitr , odorless, granular Page 116 ARGUS 418-023 PAGE 2 .......... F m POflST:......... ........ . LIMITS LEL: ....... . F m L E LIMITS llEL:. ..... AlrrortinnIW TEMPERATURE:. Won-F-ble N/A NlA N/A ~ ~ S n I l w 3 u0 : Mtm, carbon dioxide, W mk.l,Foam SPECUL FIRE FIOWTING PRoCEDLREs: Herr full p r o t e c t i v e clothing, including helmot, self -contained, positive pres8ure o r pressure d r u n d brmrthing apparatus, bunker coat and pants, bands around 8m8, wirt 8nd legs, face mask, and protectivm covering ?or rxposd a n a 8 o f the head. ulusw FIRE AND MPLOSfOeY m: See Hazardous Decorpsiti#, s r c t h n f o r products of combustion. STABILITY: Stabla - I M A T I B I L I T Y mn knobm. ~ T E R I M S / c o w D ~ o NTsO AVOID: HAuRlws POLYMERIZATIffl: Hurrdout polymerization w i l l not occur. mamolls M#mposmfflPRo#IcTs: Carbon Monoxide 8nd Carbon Dioxide, Oxidmr o f Sulfur, Hydrog8n Fluoride, Toxic Vapors, Gmer o r P a r t i c u h t c s . SPILL RESPONSE: Obmrve precautions from other sections. Ventilate area. Collect s p i l l e d materi8l. Use mt sneeping compound o r m t e r t o avoid dusting. Clem up ralidua. P l r o in a closed container. REG- DISPOSAL: Incinerate in an i n d u s t r i a l o r commercial f a c i l i t y in t h e presence o f a combustible m 8 t ~ i . l . Cwburtion products w i l l include HF. Disposal r l t s r n 8 t i v e : Dispose of m s t c product in a f a c i l i t y pcrritted t o rcccpt chemical butte. ___________-_-----__--------------------------------------------------------- Abbreviations: NID - Not Deterninad N I A - Not Applicable CA - Approximately Page 117 ENVIRQlCOCTAL MTA: Not detenined. ARGUS 418-023 PAGE 3 EYE COWTACT: I r e d & t 8 l y flush eyes wfth large amounts ot wtrr. Get i u e d h t 8 medical attention. S U N c#TTAcT: mth rffect8d a n a with 808p and I4at.r. 1"AUTIoll: If s i ~ n s l 8 y . p t m s occur, I y o v e parson t o fresh a i r . I f signsltymptms continuo, call I physician. IF w : I f a m l l o m d , c i u 8 phy8ici.n M i a t e l y . Only induce vomiting a t the instruction of 8 phyrieian. Never give anything by mouth t o an unconscious person. EYE PROTECTIOW: mold eyr contact. The follouing should k worn a&na or in combination, as approprirta, t o prevent aye contact: near vented goggles. Ck8r 88fety glrrses n i t h t i d e shields. SKIN PROTECKON: Avoid skin C0nt.e. b a r 8WVWprht8 flOV86 w h m handling this material. A pair o f gloves u d o fma the folloning mllterial(3) 8m recawended: neoprene, n i t t i l a rubber. h e one o r #re of the folloninp psrsonal p r o t e c t i o n i t e a r 8s necessary t o prevent skin contact: herd covering, Eovemllt. Protective garrents (other than - _p_lov-e_r) -sh-o.u-ld-b-e_ro-de_o-f -ei-th-e-r -o-f -th-e-f-o-ll-w-in-g-m-a-te-ri-al-s:- - - - - - - - - - - - - - - - - - - - Abbreviations: NID :Not D a t e N h e d MIA Not Applicable CA - Approxinrtely Page 118 ARGUS 418-023 PAGE 4 Plain Tyvek o r coated Tyvek O r t h i l a r disposable c o v e r i l l . RE- VENlIURON: Provide appropriate 10- exhaust product i s he8t.d. Provide - appropriate local O x h 8 U 8 t v o n t f h i b n m t transfer points. Provide sufficient vantil.tan to m.intrm ai.riorw kbn mcorended exposure U n i t s . If exhaust w n t f l t i o n it not 8dequrte, use appmprhte r e s p i n t o y proteetion. RESPIRATORY PRoTEcTIOU: Avoid b m 8 t h u g drborn. m 8 t 8 r w . A m i d breathing O f dust. SeUct one ot.the toUmdng NIW approved respirators based on drbom8 EonWntr8tiOn Of C O I B t U f n M t S m d fn 8 C C O r d M C 8 d t h 0S)u rmgul8tiDns: h8lf -mask dust and D i r t mspirator, tull-?ace dust 8nd m i s t respAntot. PREVDnrON ff AccfMwlK ~ ~ I o W : Do not 08t, drfnk o r smoke when using t h i r product. Hash exposed amas thoroughly with soap 8nd M t e r . a s h hands 8tter hondUnQ 8nd b e f o r e eating. Do not ingest. RE- STORkoE: storm y . y from heat. when not in .SU Koep container d y . FIRE MR) mosmN AVOIDANCE: Nat lpplLuble. ~ o n f l 8 ~ a b l . . Keep container closed OTHER PRECWlZONUW INFORMARON: NO smoking: 9.drhg w h i l e using t h i s product can result i n c o n t u i n a t i o n of t h e tobacco and/or Roke and le8d t o the fornation o f th. h828tdOUS decomporition products mentioned h the Reactivity Data rmction of t h i n FSUS. S t a m work clothes separately f r o m other clothing, food and tobacco products. SKIN WTATIffl: L i s t e d SUbtt8nCeS indiC8t.d w i t h * Y ' under SKIN refer t o t h e p o t r n t h l contribution t o thr o v c r 8 l l exposure by the C U t m W U S r o u t e i n c l u d i n g mucous a a b n n e m d mye, e i t h e r by airborne or, mom p a r t i c u l a r l y , b_y_ d_ i-n- _c_t -c-a_n-ta-c_t_w-i-t_h-t-h-e- -s-u- -b-s-ta-n-c-e-.- - -V-e- h- -ic-l-e-s- -ca- n- -a- -lt-e-r- -s-k-i-n- -8-b-S-O-r-p-ti-O-n-.- - - - - - - - - - - Abbreviations: M I D - Not Determined M I A - Not Applicable CA - Appmximtely Page 119 m:L-7038 DEVEL-M Decnbor OB, 2000 cIcITo;(IK MPOSLIRE UMTS (continued) S- OaUR:# OF %POWE L f m f MTA: Sn Recowended Exposure Guidelines - IYOWE: Won0 Established ARGUS 418-023 PAG 5 EYE WNTACT: W e r a t e Eym I W f t 8 t b I I : S i g n 8 / S Y W t M S can i n c l u d e mdness, smiling, pain, tearing, .and h u y vision. SKIN WNTm: Contact d t h the skin during produet use i 8 not expected t o r r s u l t i n signif imnt irritatbn. 1"AUlToN: S i n N e mremxposurr, .bow rrcow8nded guidelines, u y cause: I r r i t a t i o n ( u w r mrpintoy): signslsymptms can include tormess of the nose and throat, coughing and sneezing. IF SWAObED: m y be h8rrtu.l it mallmod. Page 120 )4sDs: L-7038 D E V E l O M & MhlERIK December 06, 2000 ARGUS 418-023 PAGE 6 m, The intorration in t h i s r n t o r h l Safety Data Sheet (-) is believed t o be correct as o f the date b8u.d. Sn HUES No WRAKnES, EXPRESSED OR IWLIED, I I Y C L W Ie~u,r NQT 1m-m AWY rmxm WARRAWTV OF HERCHAHTABILfTV OR FIT"= Foil A PARTICUM PURPOSE OR COUISE of PERFOWANCE OR U S G O f TRADE. mer i s responsible f o r deterwining whether t h e 3M product is f i t f o r a particular purpose 8nd suitable t o r user's method of Ute o r r p p u t i o n . Qivsn t h o variety of factors that Cu) 8 f f O C t the use and 8wat- Of 8 PfOdUtt, $We Of J I i C h in uniquely within tho user's knowledQt and control, it i s e s s e n t i a l that the user evaluate the 3n product t o determine whether it is tit f o r a particular purpose and Sult8ble f o r user's method o f use o r application. pr0VfdO8 inforration i n electronic t o r r as I service t o its customers. Due t o the mm0t.e p o t s i b U t y that electronic t r a n r t e r may hive resulted in errors, w i t t i o n s o r alterations i n t h i s i n t o m i t i o n , 3l makes no represrntations as t o its mmpletme8s o r accuracy. I n addition, information obtained f r a 8 dat.brre m y not be as current i t the information i n the nsDS r v a i l 8 U e directly from W. . Page 121 ~ ~~ ARGUS 418-023 ATACHMENT 3 TEST SUBSTANCE PREPARATION PROCEDURE Page 122 ARGUS 418-023 ATTACHMENT 3 Protocol 418-023 Version: 418-023(05FEB 011 Page 1 of 2 TEST SUBSTANCE PREPARATION PROCEDURE Test Substance: PFBS Vehicle: Aqueous 0.1% CMC (medium viscositv) A. Purpose: The purpose of this procedure is to provide a method for the preparation of dosage suspensions of PFBS for oral (gavage) administration to rats on Primedica Argus Research Laboratories, Inc., Study 418-023. B. General Information: 1. All suspension containers will be labeled and color-coded. Each label will specify the protocol number, test substance identification, Argus batch number, concentration, dosage level, preparation date, expiration date and storage conditions. - 2. Suspensionswill be prepared: - Daily Weekly -X Approximatelyevery ten days For- days of use - By Sponsor 3. Suspensionswill be prepared at a final dosage volume of 10mUkg. 4. Safety: - X Nitrile or neoprene gloves, uniform/lab coat, goggles or safety glasses with side shields - X Dust-Mist Respirator if used in a chemical fume hood - X Half-Face Respirator if not used in a chemical fume hood - Full-Face Respirator/Positive Pressure Hood - X Tyvek Suit or tyvek apron and sleeves 5. Dosage suspensions adjusted for YOActivity/Purity or Correction Factor: - Yes - X No (Calculations based on 100%) - % Activity -YoPurity - Correction Factor 6. Sampling requirements: Cited in protocol 7. Storage: Cited in protocol Page 123 ARGUS 418-023 AlTACHMENT3 , Protocol 418-023 Version: 418-023(05FEB 011 Page 2 of 2 TEST SUBSTANCE PREPARATION PROCEDURE NOTE: Prior to test substance preparation accurately measure the required amount of the appropriate vehicle (R.O. deionized water should be used for calibration purposes) in a graduated cylinder, pour the required amount of vehicle into a beaker. Carefully mark each beaker at the meniscus. This mark will be used during the preparation to bring the test substance slurry up to volume. C. Dosage Suspension Preparation: 1. Weigh the required amount of test substance on a piece of weigh paper or into an appropriately sized mortar (see PREPARATION CALCULATIONS). 2. If weigh paper is used, transfer the test substance to an appropriately sized mortar. If necessary, grind the test substance into a fine powder. Slowly add a small amount of vehicle and grind. Continue to add vehicle slowly and grind the vehicle and the test substance together to form a fine slurry. Transfer the vehiclehest substance slurry to a marked beaker. 3. Rinse the mortar and pestle with additional vehicle to remove any remaining test substance. Transfer rinse to beaker. 4. Add additional vehicle to the beaker to bring volume up to the mark. Place on magnetic stir plate and agitate prior to and during administration andor sampling. 5. Repeat steps (1) through (4) for each concentration. Written By: I-' ' 1.2.k Clarification: 90 -Yes [see attached clarification form] InitiaVDate : I&##- > / z ~ , h Page 124 ~ ~~ ~ ~ ARGUS 418-023 APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY Page 125 ARGUS 418-023 DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 1. On 22 February 2001, 1% carboxymethylcellulose,instead of 0.1% carboxymethylcellulose,was prepared and used to make the prepared formulations that were used for dosage administration on 25 February 2001. This deviation did not adversely affect the outcome or interpretation of the study because the correct concentration of test substance was administered. 2. On 13 March 2001, day 21 of gestation, the gravid uterus was not weighed for female rats 19107 and 19183 in the 0 (Vehicle) and 1000 mg/kg/day dosage groups, respectively. T h s deviation did not adversely affect the outcome or interpretation of the study because sufficient data were collected to evaluate this parameter. All deviations are documented in the raw data. Asswiate Director M e a r c h and Study Director Page 126 ARGUS 418-023 APPENDIX E CERTIFICATE OF ANALYSIS Page 127 ARGUS 418-023 =\ CEntrE Analytical Laboratories. Inc. 3048 Research Drive &Phone: (814) 231-8032 State College, PA 16801 w.centrelab.com Fax: (814) 231-1253or (814) 231-1580 CERTIFICATE OF ANALYSIS Centre Analytical Laboratories COA Reference #: 023-067 3M Product: PFBS, Lot #5 Purity: 98.2% Test Name Specifications Purity' Result 98.2 Yo Appearance Identification NMR Metals (ICP/MS) 1. Calcium 2. Magnesium 3. Sodium* . 4. Potassium* 5. Nickel 6. Iron 7. Manganese rota1 % hpuni@ (NMR) rota1 YOImpurity &WS) rota1 % ~mpurity 'GCMS) tesidual Solvents TGA 'unity by DSC norganic Anions (IC) 1. Chloride 2. Fluoride 3. Bromide 4. Nitrate 5. Nitnte 6. Phosphate 7. Sulfate kganic Acids '(IC) 1. TFA 2. PFPA 3. HFBA 4. NFE'A lemental Analysis': 1. Carbon 2. Hydrogen 3. Nitrogen 4. Sulfur 5. Fluorine White, crystalline solid Theoretical Value = 14.2% Theoretical Value = 0.0% Theoretical Value = 0% Theoretical Value = 9.5% Theoretical Value = 50.6% Conforms Positive 1. 0.002 wt./wt.% 2. -=0.001 wtJwt.% 3. 1.142wt/wt.% 4. 8.442 wt./wt.% 5 . <0.001 wt./wt.% 6 . 0.002 wt./wt.% 7. co.001 wt./wt.% 0.9wtJwt.% 0.774 wt./wt.% None Quantified 0.02 wt./wt.% 99.69% 1. <0.015 wt./wt.% 2. 0.101 wt./wt.% 3. c0.040 wt./wt.% 4. <0.009 wt./wt.% 5 . ~0.006wt./wt.% 6. <0.006 w t J w t . % 7. ~0.078wt./wt.% 1. CO.1 wt./wt.% 2. co.1 wt./wt.% 3. co.1 wt./wt.% 4. <0.25 wt./wt.% 1. 10.4 wt./wt.% 2. 0.3 1 1 wt./wt.% 3. 3.94 W./wt.% I 4. 9.46 wt./wt.% 5. 38.1 wt./wt.% COA023-067 Page 128 Page 1 of2 ARGUS 418-023 CEntre Analytical laboratories. Inc. 3048 Research Drive State College, PA 16801 www.centrelab.com (814) 231-8032 Fax: (814) 231-1253 or (814) 231-1580 CERTIFICATE OF ANALYSIS Centre Analytical LaboratoriesCOA Reference #: 023-067 Date of Last Analysis: 08131/01 Expiration Date: 08131/03 Storage Conditions: Frozen 1-10"C Re-assessment Date: 08/3 1/03 'Purity= 100%- (Total NMR impurities, 0.90%+ Total LCMS impurities, 0.774% + Total Metal impurities, 0.004% + Total Anion Impurities, 0.101 + Residual Solvents TGA, 0.02%) Total impurity from all tests = I .799% Purity = 100% - 1.799% = 98.2% 'Potassium and Sodium are present as counterions and thus are excluded &om the purity calculation. '"FA HFBA NFPA PFPA Trifluoroacetic acid Heptafluorobutyric acid Nonafluoropentanoic acid Pentduoropropanoic acid 4Theoretical value calculations based on the empirical formula, C4F9S0,XC (MW=338.2) Prepared By: Scientist, Centre Analytical Laboratories \\a\or Date Reviewed By: 'Laboratory Manager, Centre Analytical Date Laboratories COA023-067 Page 129 Page 2 of2 ARGUS 4 18-023 APPENDIX F ANALYTICAL REPORT Page 130 ARGUS 418-023 ANALYSIS OF DOSING SOLUTIONSUSED AT ARGUS RESEARCH LABORATORY FOR SPONSOR'S STUDY NUMBERS T-7485.11, T-7485.12, AND T-7485.13 (ARGUS RESEARCH LABORATORY PROTOCOL NUMBERS:418-023P, 418-023 AND 418-021) STUDY ID: A343.1 SPONSOR STUDY NOS.T-7485.; 1 T-7485.12 T-7485.13 Southern Research Institute 2000 Ninth Avenue South P.O. Box 55305 Birmingham, AL 35255-5305 Page 131 ARGUS 418-023 SUMMARY A total of 30 formulated dose samples including blanks and vehicles ranging in concentration from 3 to 200 mg/mL were analyzed by analytical method BACG 3533 to determine the concentration of perfluorobutanesulfonate (PFBS) in the formulated mixture. All dose formulations were found to be within k 10% of the reported concentration except for four samples, 100 mg/mL dated 1/1I/O 1, 100 m g h L dated 2/23/01, 100 mg/mL date 9/4/0 1 and 200 mg/mL dated 1/11/01. These three samples were found to have measured concentrations of 88.9 %, 86.3 %, 76.6% and 76.8 % of target values, respectively. Page 132 KEY PERSONNEL Raymond G. York, Ph.D., D.A.B.T. Study Director Argus Research Laboratories . James D. Johnson, M.S., MBA Manager Bioanalytical Chemistry Group Gregory S. Gorman, Ph.D. Staff Chemist Bioanalytical Chemistry Group Lara Cook, M.S. Research Associate I1 Bioanalytical Chemistry Group ARGUS 418-023 Page 133 ARGUS 4 18-023 1. OBJECTIVE The objective of this study was to determine the dose concentration of PFBS in the supplied dosing solutions received from the sponsor. 2. SAFETY All necessary procedures to ensure safety of the analysts were based on information contained in the Material Safety and Data Sheets (MSDS), provided by the producer of the test article and the study director. 3. Compliance The study described in this final report was conducted in accordance with the EPA Good Laboratory Practice Regulations (40 CFR Part 160 and 792), OECD Principles of Good Laboratory Practice [C(97)186/Final]. For studies 418-023 and 418-023P Good Laboratory Practice Standards (1984) for MAFF 59 Nohsan No. 3850 were met. The final report accurately reflects the raw data obtained during the performance of the study. There were no adverse circumstances that affected the quality or integrity of the study. 4. EXPERIMENTAL 4.1 Analytical Procedures The sample preparation and analysis procedures as described in the analytical method BACG 3533 were employed for all analyses. Each sample was allowed to warm to room temperature and was then vortexed well before being sampled. An aliquot was taken from each and diluted as described in the method. (Note: the 66.7 mg/mL and higher concentrated samples often were suspensions and the entire sample volume in some cases were diluted to the target range). Multiple calibration curves were prepared over a concentration range of 500 to 10,000 ng/mL and analyzed along with the samples as described in the method. The correlation coefficient for each curve was greater than 0.9982. 4.2 Results The results of the analysis are presented in the Tables 1-111corresponding to the sponsors study number at the end of the report. Page 134 ARGUS 418-023 5.0 Conclusion A total of 30 dose formulation samples ranging in concentration from 3 to 200 mg/mL were analyzed by BACG 3533. All samples except for four, 100 mg/mL dated 1/11/01, 100 mg/mL dated 2/23/01, 100 mg/mL dated 9/4/01 and 200 mg/mL dated 1/11 were found to be withln k 10 % of the reported concentration. Page 135 ARGUS 418-023 Table I Dose Formulation Analysis of PFBS in 0.1% CMC Sponsor Study Number T-7485.13 I Small amount of material detected in the undiluted sample is considered insignificant when compared to amount found in diluted samples and is believed to be the result of carry-over from a previously run standard injection. `Average of two results. Page 136 ARGUS 418-023 Table I1 Dose Formulation Analysis of PFBS in 0.1% CMC Sponsor Study Number T-7485.12 Page 137 ARGUS 418-023 Table I11 Dose Formulation Analysis of PFBS in0.1% CMC Sponsor Study Number T-7485.11 Page 138 6.0 Approvals ARGUS 4 18-023 Research Associate I1 Bioanalytical Chemistry Group Staff Chemist Bioanalytical Chemistry Group Tina Rogers, Ph.D. Director Safety Assessment Department Date /'?faa w ? Date Page 139 ARGUS 418-023 Quality Assurance Statement Final Report On Analysis of Dosing Solutions Used at Argus Research Laboratory for Sponsor's Study Numbers T-7485.11, T-7485.12, and T-7485.13 (Argus Research Laboratory Protocol Numbers: 418-0231', 418-023, and 418-021) A343.1 Listed below are the phases andor procedures performed by Southern Research Institute that were inspected and audited by the Quality Assurance Unit during the study described in the report. Findings were reported to the study director and management periodically. Ph asedProcedures Inspection/ Audit Date Date Management Notified Date Study Director Not fled' Protocol Review I 5114101 1 5/14/01 1 1/18/02 1 Dose Formulation Concentration Analysis 5114101 6 /14101 1/18/02 I 1 II I I 1 Data Audit and Draft 12/3/01-12/6/01; Report Review I/ 14102-1117/02 1/18/02 1/18/02 1 FinalReport I 1 I 1/30/02 1 I 1/30/02 1 I I 1/30/02 I I I I I The study director is off-site. 2.-Quality AssuranceIQuality Control / Date / L-- Page 140 ARGUS 4 18-023 APPENDIX G ENVIRONMENTAL AND HUSBANDRY REPORTS Page 141 ARGUS 418-023 ARGUS Temperature and Relative Humidity Report Location: Room 03 Protocol Number: 418023 Range of Dates: 13-Feb-2001 08:OO to 23-Feb-2001 1 6 5 9 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64F to 79F 11 248.75 249 Relative Humidity 30% to 70% 11 248.75 249 Mean (2 SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 70.8 73.1 71 .O 66.5 249 0 0 (k 1.3) (100.0) (0.0) (0.0) 36.6 50.2 35.7 31.2 249 0 0 (* 3.2) (100.0) (0.0) (0.0) Report Generated: 15-Mar-2001 at 14% COMMENTS: , REVIEWED BY: DATE: :umulative by Location (vMay-21-1999) Page 142 ARGUS 418-023 ARGUS Temperature and Relative Humidity Report Location: Room 20 Protocol Number: 418023 Range of Dates: 23-Feb-2001 16:OO to 15-Mar-2001 1659 Target Range: Species: Rat I Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64'F to 79'F I 21 I 470.5 I 477 Relative Humidity 30% to 70% 21 478.5 477 Mean (k SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High ("h): Number of Points Low (%): ~~ ~ Report Generated: 15-Mar-2001at 14:46 71.1 72.0 71.1 69.8 477 0 0 ~~ (i0.4) (100.0) (0.0) (0.0) ~~ 51.4 65.7 52.0 31 .O 477 0 0 (* 6.3) (100.0) (0.0) (0.0) COMMENTS: REVIEWED BY: / . DATE: umulative by Location (vMay-21-1999) Page 143 Certified Papers Retrieval Return to Certified Analysis Retrieval I Product Code: Product Desc: Lab Number: Lot Code: Entered: 5002M CERTIFIED RODENT DIET MEAL LOO25623-1 OCT 03 00 3A 10/5/00 Assay PROTEIN Analysis- 21.51-1 5.64j./"I ARGUS 418-023 Page 1 of 2 Aldrin Beta-BHC ODE Delta-BHC Endrin Heptachlor Lindane Mirex LESS THAN 0.02 LESS THAN 0.02 LESS THAN 0.02 LESS THAN 0 02 LESS THAN 0.02 LESS THAN 0.02 LESS THAN 0.02 LESS THAN 0.02 Alpha-BHC Chlordane DDT Dieldrln HCB Heptachlor Epoxlde Methoxychlor PCB I LESS THAN 0 02 LESS THAN 0 02 LESS THAN 0 02 LESS THAN 0 02 LESS THAN 0 02 LESS THAN 0 02 LESS THAN 0.02 LESS THAN 0.1 5 http:liwwv. labdiet.com/certified/pwa-spc002.asp Page 144 -- ... __ 12/10/2000 Certified Papers Remeval ARGUS 4 18-023 Page 2 of 2 [Aflatoxin IlAflatoxins ]!LESS THAN 5 PPB No notes. For additional information, please Contact: -- 1) Michael J. Murphy at (314) 982-2383 for assay methodology -- 2) Dr. Dorrance Haught at (314) 768-4362 for nutritional interpretation - 3 ) Richmond, IN ManufacturingPlant at (765) 962-9561 all other questions The iemt '"Less Than" IS used 10 s t J i l ~ f y the loucr liniil ofquaittiinrioit o i the procedure uiioer ilic condtiionr cniDIo,ea The use 01 the term 'Less Titan" docs UOIintply t h x traces o i a n i l ? t e*ere present. 12/20/2000 ARGUS 4 18-023 Page 146 ARGUS 4 18-023 Pqp 1 of2 Smpk NO.: uKX28475-2 R s a i v d : 11/3o/oo Rcpor(ed: lY14/00 PRKR FrEW?TAI FEBR ASP CDP CAF PB HGSP PP SE 20.8 5.92 3.99 a200 0.053 0.977 0.139 Q).m 0.659 0.159 REVIEWED BY: DATE Page 147 ARGUS 418-023 ORCP RSPB AFK Rsrult 4.02oO 4.0200 4.0200 4,m CD.Mo0 4.m 4.0200 a0200 4.Moo 4.0200 aom 4.0200 4.0200 4.0200 4.02oo 4.0200 Q.o200 4.Ou)O 4.0200 4.0200 4.MM 4.m 4.moo 4.0200 4.150 4.00 Page 148 ARGUS 418-023 Lancaster Laboratories Where quality is a science. Laneaster Laboratories Sample No. Ww 3 4 0 2 2 9 0 collecied:06/20/2000 09:30 by EA Submitted: 06/20/2000 17:OO Reported: 07/18/00 at 05:50 AM Discard: 8/18/00 Point f l Analytical Lab Grab Water 9 0 5 Sheehy Bldg. B Sample Account Number: 02423 Prirnedica Argus 9 0 5 Sheehy Drive, Sie. A Hozsham PA 19044-1297 Pl--- CAT No. 00224 00228 00368 0150.i 01506 07322 Ol505 Analysis Namm Chloride Sulfate ?:-crate Nicrogcr: Fluoride Nitrice Nicroger: Orcho-phosphace Bromide 00178 00453 00454 00455 00469 006700478 03638 01902 01903 01904 a1905 C1906 01907 ai908 ai909 01910 0191: ai91?. Oi9-3 01914 01915 01916 0191: 01918 Cl919 Pesc:cides/P~3's Ln Maces G a m a BHC - Lrndane liep:achlor Aidrin Di e1dr1.1 L.?dZZ.-. DSS Ezr'rrx Ald=h::dAlpha BHZ ae=a BKC Delta BHC ileprachlor Epoxide DDZ DCD Chlordane Toxaphene E2dosulfan Z Ecdosulfan :I - Endosulfan Sulfate PCB 101 6 pca-122: pca - it3I - ~ c a125 z - PC3 1248 PCB-1254 ?C9- i26 0 CAS Numbrr 16887-00-6 14808-79-8 14797-55-8 16984-48-8 14797-65-0 13265-43-2 n.r. u Rrcrivmd Rosult c 2.0 c 5.0 c 0.50 c 0.50 c 0.50 5.0 c 2.5 h Rrcrivrd L i m i t of Purrrtit r t i o n 2.0 5.0 0.50 c . sc 0.50 5.0 2.5 58-89-9 76-44-8 309-00-2 60-57-1 72-20-8 SO-29-3 7421-93-4 319-84-6 319-85-7 3ig-wi-8 1024-57-3 - 7 2 - 5 5 - 9 72 -54 8 57-74-9 8001-35-2 959-98-8 33213-65-9 1031-07-8 12674-11-1 11104-28-2 11141-16-5 53469-21-9 12672-29-6 11097-69-1 11096-82-5 < 0.0096 .a 0 . 0 0 9 6 c 0.0096 < 0.0096 c 0.0096 0.0096 < 0.095 0.0096 < 0.0096 < 0.0096 < 0.0096 c 0.0096 c 0.0096 0.29 c 3.8 c 0.0096 c 0.0096 c 0.029 < 1.0 < 1.0 < 1.0 c 1.0 1.0 c 1.0 c L.0 0.0096 0.0096 0.0096 0.0096 0.0096 0.0096 0.096 0,0096 0.0036 0.0096 0.0096 0.0096 0.0096 0.29 3.8 0.0096 0.0096 0.029 1.0 1.0 1.0 1.0 1.0 1.0 1.0 ug/l ug/1 ug/l ug/l ug/ 1 ug/ 1 ug/l ug/l ug/1 ug/l ug/l ug/l ug/ 1 ug/l ug/l ug/l ug/l ug/l ug/l ug/l ug/l ug/l ug/l ug/l ug/l Page I of 2 Dilution ?ac t o r 5 5 5 5 5 5 5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Page 149 ARGUS 418-023 LancasterLaboratories Where quality is a science. Lancarter Laboratories Saaupl. N o . W W 3402290 Collected:06/20/2000 09:30 by Submitted: 06/20/2000 17:OO Reported: 0 7 / 1 8 / 0 0 a t 0S:SO AM Discard: 8/18/00 Point #1 A n a l y t i c a l Lab Grab Water 905 Sheehy Bldg. B Sample Account Number: 02423 Primedica Argus 905 Sheehy Drive, Ste. A Horsham PA 1 9 0 4 4 - 1 2 9 7 PI.--- CAT NO. AnSlysiS Name accuracy at a batch level CAS Numbmr U Rmcmivmd R.ault Aa Rmemivrd L M t of Quaatitation Units The beta-BHC and endosulfan sulfate recoveries are outside the QC limits for the X S D . Since the recoveries were high and no beta-EH: or endosulfan sulfate was detected in the sample. the results were reported. 01856 Herbicides in water 01857 01858 05286 05287 05288 05289 05290 05291 05292 05291 08103 2.4-D 2.6.5-TP 2.4.5-T Dalapon Dinores Dicarrba KC7P MCPA 2.4-DP (Drchlorpropl 2.4-DB Pentachlorophenol 94-75-7 91-72-1 93-76-5 75-99-0 88-85-7 1918-00-9 -9 3 - 6 5 - 2 94 74-6 120-36-5 94-82-6 87-86-5 < 0.48 c 0.048 0.048 < 1.2 4 0.24 < 0.29 < 190. c 190. c 0.48 c 0.48 < 0.05 0.48 0.048 0.048 1.2 0.24 0.29 190. 190. 0.48 0.48 0.05 Commonwealch of Pennsylvania Lab Cercificacion No. 36-037 ug/ 1 ugll ug/ 1 ug/l ug/l ug/ 1 ug/ 1 ug/1 ug/l us/ 1 ug/ 1 Page 2 of 2 Dilution hetor 1 1 1 1 1 1 1 1 1 1 1 Page 150 ARGUS 418-023 qlp Lancaster Laboratories Wherequalily is a science. Lancaster Laboratories Sample No. WW 3542351 Collected:01/26/2001 08:40 by EA Submitted: 01/26/2001 16:OO Reported: 02/06/01 at 05:22 AM Discard: 3/9/01 905 Formulation Lab Point t 2 Grab Semi-Annual Water Sample Accounf Number: 02423 Prisedica nrgus 5 1 Union Street Worcester MA 01608 Page I of 1 905-2 CAT No. 00224 00228 00368 01504 015?i 07322 01505 Analyrir.m&N Chloride Sulfate Nitrate Nitrogen Sluorxie ::: zrlZE Nitrogen 0r;ho-phosphate Sramide 00178 00453 00454 00455 0045'3 00477 50479 -.*-. ".,A: 01502 01903 01904 01905 CL3CS 01907 01908 01909 01910 01.911 01912 01911 01914 01915 01515 01317 C1319 51919 Pescrcides/PCB's in Water Gamma 8HC - Lindane Heptachlor Aldrin Pirl;trin Endrin 331 Z:.iziz .;ki=>15= Alpha JXC :=:a a x Oelca 3HC He?cacr.lor SpoxSd. 339 335 Cklo-dane Taxapnene E.-.losulan I Endosulfan I1 Endosulfan Sulfate Ita- l o i s - PC3-1221 PCI 122.2 - ?C3-1242 ?C3 i2i8 1 3 ~ - 2 54 IC3~ 12; 0 UIS Number 16887-00-6 14808-79-9 11797-55-9 1698+-48-8 :;-+--<j-: ii255-44-2 n.a. As Rmceivid Reaul t < 2.0 c 5.3 < 0.50 c 0.jJ < :.:: c 5.0 c 2.5 A s Received Limit O f Quantication 2.0 5.0 0.50 0.50 5.52 5.3 2.5 Unitr mg: 1 nq/1 3q/1 my/? nq, 1 3q/I m3l I Dilution Factor 5 5 5 5 5 5 58-89-9 76-44-8 309-00-2 53-57-1 72-20-8 53-29-3 . - - - ; 3~ - i 119-a4-6 319-35-7 319-86-8 '024-57-3 72-55-3 71-j4-3 57-74-9 3001-35-2 353-98-8 31213-65-9 1011-07-e 12674-11-2 11104-28-2 >:141-15-5 51469-21-5 11572-19-5 11537-65-> 11035-32-5 < 0.0096 c 0.0095 c 0.0036 < 0.0096 < 0.0095 c 5.2035 c :.:35 < 0.5056 c 0.2036 c 0.0096 < 0.0035 c :..I35 c 0.3095 c 0.29 < 3.3 c 0.0096 c 0.0096 ' 5.529 < 1.0 < 1.0 c 1.2 c 1.3 < 1.3 c l.J c 1.3 3.0096 o.cs>; 0.3095 0.0096 0.0095 0.3095 C.396 0.0095 9.3035 0.3036 0.0096 3 539s 3.3096 0.23 3.8 0.3355 0.0096 0.029 1.0 1.0 1.0 1.5 1.0 1.0 1.0 uq/l uq/ 1 1 U9/l ug/l , 1 uq/ 1 - ug: L ..-,. -9, 1 1 q/l ug/1 1 L.911: uq; 1 1 Ug;l 1 uq/ 1 1 ugil 1 ugi1 1 uq/ 1 1 ug/l 1 u9/l 1 ugil 1 uq/l uq/l 1 ugi 1 1 ug/l ug/1 ug/1 1 Uncaster boorarorle5. Inc. 2625 New Uolland Pike m BOX i z u s LancaRer. PA 17605-2425 777-616-23W Fax: 717-656-2581 2116 Rev. 9/111w Page 151 ARGUS 418-023 Lancaster Laboratories where pahfyisa science. Lancaster Laboratories Samplo No. WW 3 5 4 2 3 5 1 Collected:01/26/2001 08:40 by EA Submitted: 01/26/2001 16:OO Reported: 02/06/01 at 05:22 AM Discard: 3/9/01 905 Formulation Lab Point f Z Grab Semi-Annual Water Sample A C C O U ~ C Number: 02423 Prirnedica Argus 57 Union Strest Worcester HA 01608 905-2 CAT No. 01856 Analyaia Nu. Herbicides in Water 01857 01858 OS286 05237 05288 OS289 OS290 OS291 OS292 05293 08103 2.4-0 2.4.5-TP 2,4,S-P Calapcn Drnoseb Dicamba MCPP MCPA 2.4-DP (DichlorpropJ 2.4-08 Pentachlorophenol CAS Number A. R8cmiv.d Rmmult 34-75-7 93-71-1 93-76-5 75-33-0 88-as-7 1918-00-9 93-6 j - 2 94-74- i 120-35-5 94-82-6 87-85-5 c 0.48 c 0.048 c 0.048 c 1.2 < 0.21 < 0.29 < 190. c 190. c 0.48 c 0.48 c 0.05 C;monueal:h of Pennsylvania Lab C-rcificacion NO. 36-037 rU X a c r i v e d L i m i t of gumtitation Units 0.48 0.048 0-- ..--048 0.24 0.29 190. 130. 0.48 0.48 0.05 Ug/ 1 ugI1 Ug/l ug/i ug/l u9/i u9: u9/l ug/1 ug/ 1 ug/ 1 Page 1 of 2 Dilution Factor 1 1 1 1 I 1 1 . I ME e E m u s t e r Laboratories. Inc 1425 New Holland Pike W Boa 12425 bncasrer. PA 1?6Q5*24l5 117-6SCZ300 Fax: 717-656-16881 Page 152 ARGUS 418-023 ANALYTICAL LABORATORIES, INC. P.0. Bax 319 cHALmm, PA 18914 (215) 723-6466 customer: Argus Research, Inc. 905 Sheehy Dt., Bldg. A Horsham, PA 19044 Attn. Dena Lebo Attn : 215-443-8710 FAX t i : Sample number: 2337-OlA Date sampled : 02/14/01 Time sampled : 1515 Dace received: 02/14/01 Sampled by : EM Water Safety Classification: No Coliform bacteria were detected. Therefore, the water supply, at the time o f sampling, meets the &PA and DEP drinking water standards for this parameter. Coliform Bacteria, councs/100 m l c- Noncoliform Bacteria, counts/100 m l Total Chlorine, mg/l <l c1 < 0-1 ~ l rlesults which are outside the "Maximum Contaminant Level" (MCL) established under the "Safe Drinking Water Act" are marked by asterisks ( * * I . Symbol key : c - less than TNTC - too numerous to count mg/l - milligram/llcer ( > 200) PA DEP ciO9-332 Maryann E. Fedock/ President Page 153 ARGUS 4 18-023 ANALYTICAL LABORATORIES, INc. P.0. BOX 319 cm.Lmm, PA 18914 (215) 723-6466 PRPQBT Customer : Argus Research, Inc. 905 Sheehy Dr., B l d g . A Horsham, PA 19044 Attn. Dena Lebo Attn: 215-443-8710 FAX # : Sample source: Rm. 5 @ D L A%& Sample number: 2337-01C Date sampled : 02/14/01 Time sampled : 1515 Date received: 02/14/01 Sampled by : gM ANALYTICAL RESULTS Water Safety Classification: No Coliform bacteria were detected. Therefore, the water supply, at the time of sampling, meets the EPA and DBP drinking water standards for this parameter. Parameter ICL Coliform Bacteria, counts/100 ml e1 Noncoliform Bacteria, COuntS/100 d Total Chlorine, mg/l Result c1 <I. 0.1 All results which are outside the "Maximum Contaminant Level" (MCL) established under the "Safe Drinking Water Act" are marked by asterisks ( * * ) . Symbol key: < - l e s s than TNTC - too numerous to count ( > 2 0 0 ) mg/l - milligram/liter PA DEP 809-332 m- I -2y4 */-/& Maryann E. Fedock/ President Page 154 ARGUS 418-023 ANALYTICAL LABORATORIBS, P.O. Bar 319 CElUZONT, PA 18914 (2154) 723-6466 INC. Customer: Argus Research, InC. 905 Sheehy Dr., Bldg. A Horsham, PA 19044 Attn. De- Lebo Attn: 215-443-8710 FAX # : Sample number: 2337-om Date sampled : 02/14/01 Time sampled : 1515 Dace received: 02/14/01 Sampled by : EM & Sample source: Analytical 3k ~ L SSA C C A Y b c / y ~ d / ~ a / c / MALITICAL RESULYS Water Safety Classification: No Coliform bacteria were detected. Therefore, the water supply, at the time of sampling, meets the BPA and DBP drinking water standards for this parameter. Parameter Coliform Bacteria, counts/100 ml c- Noncolifonn Bacteria, counts/100 ml Total Chlorine, mg/l Result cl <1 < 0.1 All results which are outside.che "Maximum Contaminant Leveln (MCL) established under the " S a f e Drinking Water Ac:" are marked by asterisks ( * * I . Symbol key : c - less than `ITJTC - too numerous to count mg/l - milligram/liter ( > 200) PA DEP # 0 9 - 3 3 2 *Z--e&& L9d Maq&nn E. Fedock/ President Page 155 ARGUS 418-023 AwLGYTIcat LABORATORIES, P . O . Box 319 CBALWNT, PA 18914 (215) 723-6466 INC. Customer: Argus Research, Inc. 905 Sheehy Dr., Bldg. A Horsham, PA 19044 Sample number: Date sampled : Time sampled : Attn. D e M -0 Attn: 21s-443-8710 Date received: Sampled by : FAX #: Sample source: Chem. L a b 'IDS F &hc& 1 b &-jLy+7, r'vc ANALYTICAL RESULTS 4135-01B 03/07/01 1430 03/08/01 PO Water Safety Classification: No Coliform bacteria were detected. Therefore, the water supply, at the time of sampling, meets t h e BPA and DEP drinking w a t e r standards for this parameter. Coliform Bacteria, counts/100 m l <1 Noncoliform Bacteria, counts/100 m l Total Chlorine, mg/l cl <1 c 0.1 A l l results which are outside the "Maximum Contaminant Level" IMCL) established under the "Safe Drinking Water Act" are marked by asterisks ( * e ) . C Symbo- l l key ess : than TNTC - too numerous to count mg/l - milligram/liter (> 200) FA DEP f c O 9 - 3 3 2 *-a- 4-J-.%A Marya'rin E. Fedock/ President Page 156 ARGUS 418-023 ANALYTICAL WLBORATORIBS, P . O . Box 319 cBAT;FoNT, PA 18914 (215) 723-6466 INC. Customer: Argus Research, Inc. 905 Sheehy Dr., Bldg. A Horsham, PA 19044 At-. Dena Lebo Attn: 215-443-8710 FAX # : Sample number: 4135-01C Date sampled : 03/07/01 Time sampled : 1430 Date received: 03/08/01 Sampled by : PO ANALYTICAL RESULTS Water Safety Classification: No Coliform bacteria were detected. Therefore, the water supply, at the time of sampling, meets the BPA and DEP drinking water standards for this parameter. Parameter Coliform Bacteria, counts/100 m l e1 Noncoliform Bacteria, counts/100 m l Total Chlorine, mg/l <1 e1 < 0.1 All results which are outside the "Maximum Contaminant Level" (MCL) established under the "Safe Drinking Water Act" are marked by asterisks ( * * ) . Symbol key : C - less than TNTC - too numerous to count ( > 200) m g / l - milligradliter PA DEP t 0 9 - 3 3 2 Maryann E. Fedock/ President Page 157 ARGUS 418-023 ANALYTICAL LABORATORIES, INC. P.0. Box 319 CEALPONT, PA 18914 (215) 723-6466 SIS REPORT Customer: Argus Research, Inc. 905 Sheehy Dr., Bldg. A Horsham, PA 19044 Attn. D e n a Lebo Attn: 215-443-8710 FAX # : Sample number: 4135-01D Date sampled : 03/07/01 Time sampled : 1430 Date received: 03/08/01 Sampled by : FO Sample source: Room #5 ANALYTICAL RESULTS Water Safety Classification: NO Coliform bacteria were detected. Therefore, the water supply, at the time of sampling, meets the EPA and DBP drinking water standards for this parameter. Parameter Mal Result Coliform Bacteria, counts/100 ml <1 Noncoliform Bacteria, counts/100 m l Total Chlorine, m g / l s c1 cl 1.5 A l l results which are outside the "Maximum Contaminant Levell' (MCL) established under the "Safe Drinking Water Act" are marked by asterisks ( * * I . Symbol key: C - less than TNTC - too numerous t o count mg/l - milligram/liter PA DE? 409-332 (> 200) Ma&hn E. Fedock/ President Page 158 ARGUS 418-023 Explanation of Symbols and Abbreviations The followingdefines common symbols and abbreviatiansused in m p o ~ n gtechnical data: N.D. TNTC IU umhoslcm C CaI meq 9 ug mi m3 none detected Too Numerous To Count International Units micromhodcm degrees Celsius (diet) calories milliequivalents gramw microgram(s) milliliter(s) cubic meter(s) BMQL MPN CP Units NN F Ib. kg mg I ui fib >5 umlml Below Minimum Quantitation Level Most Probable Number cobattchloroplatinateunits nephelometricturbidity units degrees Fahrenheit pounw kilogram(s) milligram(s) liter(s) rnicroliterls) fibers greater than 5 microns in length per ml C > PPm less than - The number following the sign is the limit of auantitation,the smallest amount of analyte which can be reliably determined using this specific test. greater than parts per million - One ppm is equivalent to one milligram per kilogram (mglkg), or one gram per million grams. For aqueous liquids, ppm is usually taken to be equivalent to milligrams per liter (mg/l), because one liter of water has a weight very close to a kilogram. For gases or vapors, one ppm is equivalent to one microliter of gas per liter of gas. PPb Dry weight basis parts per billion Results printed under this heading have been adjusted for moisture content. This increases the analyte weight concentrationto approximatethe value present in a similar sample without moisture. U.S. PA data qualifiers: Organic Qualifiers A B C D E J N P U X,YZ TIC is a possible aldolcondensabon product Analyte was also detected in the blank Pesticideresult confirmed by GC/MS Compound quantitated on a diluted sample Concentratton exceeds the calibration range of the instrument Estimated value Presumptiveevidence of a compound (TICSonly) Concentration difference between pnmary confirmation columns 225% Compound was not detected Defined in case narrative Inorganic Qualifiers B Value is <CRDL. but i l D L E Estimated due to interference M Duplicate injection precision not met N Spike sample not within control limits S Method of standard additions (MSA) used for calculation .U Compound was not detected W Post digestion spike out of control limits Duplicate analysis not within control limits + Correlation coefficient for MSA <0.995 Tests results relate only to the sample tested. Clients should be aware that a critical step in a chemical or microbiological analysis is the collection of the sample. Unless the sample analyzed is truly representativeof the bulk of material involved, the test results will be meaningless. If you have questions regarding the proper techniques of collecting samples, please contact us. We cannot be held responsiblefor sample integrity. however. unless sampling has been performed by a member of our staff. This report shall not be reproduced except in full, without the written approval of the laboratory. WARRANW AND LIMITS OF LIABILITY - In accepting analytical work, we warrant the accuracy of test results for the sample as submitted, THE FOREGOING EXPRESSWARRANTYIS EXCLUSIVEAND IS GIVEN IN LIEU OF ALL OTHER WARRANTIES, EXPRESSEDOR IMPLIED. WE DISCLAIMANY OTHER WARRANTIES. EXPRESSEDOR IMPLIED, INCLUDINGA WARRANTY OF FITNESSFOR PARTICULAR PURPOSEAND WARRANN OF MERCHANTABILIN. IN NO EVENTSHALL LANCASTER LABORATORIESBE LIABLEFOR INDIRECT,SPECIAL. CONSEQUENTIAL.OR INCIDENTALDAMAGESINCLUDING,BUT NOT ' IMITEDTO, DAMAGESFOR LOSS OF PROFITOR GOODWILLREGARDLESSOF (A) THE NEGLIGENCE(EITHERSOLE OR JONCURRENT) OF LANCASTERLABORATORIESAND (8) WETHER LANCASTERLABORATORIESHASBEENINFORMEDOF THE POSSIBILITY OF SUCH DAMAGES. We accept no legal responsibility for the purposes for which the client uses the test results. No purchase order or other order for work shall be accepted by Lancaster Laboratories which indudes any conditions that vary from the Standard Terms and Conditions of LancasterLaboratoriesand we hereby object to any confliing terms containedin any acceptance or order submitted by client. Page 159 ARGUS 4 18-023 APPENDIX H QUALITY ASSURANCE STATEMENT Page 160 ARGUS 418-023 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (2 15) 443-87 10 Telefax: (215) 443-8587 QUALITY ASSURANCE STATEMENT Argus Protocol: 418-023 Sponsor's Study Number: T-7485.12 Study Director: Raymond G. York, Ph.D., DAJ3T The protocol, critical phases, raw data and final report were inspected by the Quality Assurance Unit (QAU), to assure conformance with: U.S. Environmental Protection Agency. Toxic Substances Control Act (TSCA) Good Laboratory Practice Standards; Final Rule. 40 CFR Part 792. U.S. Environmental Protection Agency. Federal Insecticide, Fungicide and Rodenticide Act (FIFRA) Good Laboratory Practice Standards; Final Rule. 40 CFR Part 160. Organization for Economic Cooperation and Development (1998). The Revised OECD Principles of Good Laboratory Practices [C(97)186/Final]. Japanese Ministry of Agriculture, Forestry and Fisheries (1984). Good Laboratory Practice Standards. 59 NohSan No. 3850. The undersigned indicate that the report is an accurate representation of the raw data. Data provided by the Sponsor or a subcontractor were not audited by the Primedica Argus Quality Assurance Unit. Page 161 ARGUS 418-023 The QAU inspection and report audit dates are listed below: Inspection Phase Insuection Date(s) Date(s) Findings Date(s) Findings Submitted to Study Submitted to Director Management Protocol 03 FEB 01 16 FEB 01 Test Substance Preparation 08 MAR 01 Test Substance Administration 08 MAR 01 Caesarean-Sectioning 15 MAR 01 In-Life Data 02, 12and 14 APR 01 Formulations Data 17 APR01 & 05 MAY 01 Report Text 22-24,26 MAY 01 31 MAY 01 Report Tables 25-31 MAY 01 Revised Report 23 JAN 02 03 FEB 01 16 FEB 01 08 MAR 01 26 MAR 01 17 MAR 01 14 APR 01 06 MAY 01 26 MAY 01 31 MAY 01 31 MAY 01 23 JAN 02 03 EEB 01 16 FEB 01 08 MAR 01 26 MAR 01 17 MAR 01 14 APR 01 06 MAY 01 26 MAY 01 31 MAY 01 31 MAY 01 23 JAN 02 ............../..f.i.A...q..R...o..a.... Matthew J. Vaneman, B.S. Date Manager, Regulatory Compliance Date &ality Assurance AssociateiTeam Leader and Principal Auditor Page 162