Document 6Brnkb6EOj9XazBQOo8zQn7L9

FINAL REPORT PROTOCOL 418-015. ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.14 EINAL REPORT DATE: 23 JULY 1999 01001 PROTOCOL 418-015 ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS INRATS SPONSOR'S STUDY NUMBER: T-6295.14 TABLE OF CONTENTS SUBJECT PAGE I. SUMMARY AND CONCLUSION 2] A Methods 1 B. Results 3 C. Conclusion [" I. DESCRIPTION OF TEST PROCEDURES 11 A. Conduct of Study 11 A. Sponsor 11 A2. Testing Facilty [=] A3. Study Number 1-1 A4. Sponsor's Study Number 1-1 AS. Purpose of the Study 11 AS. Study Design 1 i 01002 SUBJECT AT. Regulatory Compliance A8. Ownership of the Study A9. Study Monitor A10. Altemate Study Monitor A11. Study Director A12. Technical Performance A13. Report Preparation A.14. Report Review A15. Date Protocol Signed A.16. Dates of Technical Performance AA7. Records Maintained B. Test Aticle Information B.1. Description B82. LoBatch Number B3. Date Received and Storage Conditions B.4. Special Handling Instructions B5. Analysis of Activity C. Vehicle Information C.1. Description C2. LotNumbers C3. Dates Received, Source and Storage Conditions Ca. Special Handing Instructions. i PAGE 1 2 2 1-2 2 2 12 2 2 3 3 3 1-3 1-3 4 1-4 14 4 4 14 14 14 01003 SUBJECT C5. Analysis of Purity D. Test Article Preparation and Storage Conditions D.1. Sample Information D.2. Analytical Results E. TestSystem EA. Species E2. Strain E.3. Supplier (Source) Ed. Sex ES. Rationale for Test System E6. Test System Data E.7. Breeder Male Rat Data ES. Method of Randomization E.9. System of Identification F. Husbandry FA. Research Facility Registration F2. Study Rooms F3. Housing F4. Lighting F5. Sanitization F6. Feed F7. Feed Analysis iii PAGE 5 Is 1-5 1-5 II-5 5 1-6 1-6 1-6 6 1-6 1-6 6 7 wr 7 wr wr 1-8 1-8 8 8 0100% SUBJECT F.8. Water F.9. Water Analysis F.10. Bedding F.11. Bedding Analysis G. Methods G.1. Dosage Administration G2. Rationale for Dosage Selection G.3. Routeof Administration G4. Rationale for Route of Administration G5. Frequency of Administration G6. Length of Study G.7. Method of Study Performance G8. Pharmacokinetic Sample Collection G.9. Gross Necropsy G.10. Statistical Analyses n. RESULTS A. Mortality, Clinical and Necropsy Observations A. Mortality A2. Clinical Observations A.3. Necropsy Observations B. Body Weights and Body Weight Changes B.1. Precohabitation v PAGE 1-8 8 ie ne 1-9 Ire iro 1-10 1-10 1-10 1-10 1-11 12 I-13 1-14 n-1 n-1 1 1 n-1 1 1-1 01005 SUBJECT PAGE B2. Gestation m2 B3. Lactation 2 C. Absolute (glday) and Relative (g/kg/day) Feed Consumption Values Ill-2 Cu. Precohabitation nz C2. Gestation 2 C3. Lactation "3 D. Natural Delivery and Litter Observations "3 E. Clinical Observations from Birth to Day 21 Postpartum and Necropsy Observations "3 REFERENCES ns APPENDIX A - REPORT FIGURE Figure 1. Body Weights -- Fo Generation Female Rats A APPENDIX B - REPORT TABLES Table 1. Clinical Observations - Summar-y Fo Generation Female Rats B-1 Table 2. Necropsy Observations - Summar-y Fo Generation Female Rats B4 Table 3. Body Weights - Precohabitatio-n Summar-y Fo Generation Female Rats B-5 Table 4. Body Weight Changes - Precohabitation - Summary Fo Generation Female Rats B6 Table 5. Matemal Body Weights - Gestation - Summary Fo Generation Female Rats B7 Table 6. Matemal Body Weight Changes - Gestation - Summary Fo Generation Female Rats Bo v C1006 SUBJECT Table 7. Table 8. Table 9. Table 10. Table 11. Table 12. Table 13. Table 14. Table 15. Table 16. Table 17. Table 18. Table 19. Table 20. PAGE Matemal Body Weights - Lactation - Summary - Fo Generation Female Rats B-10 Matemal Body Weight Changes - Lactation - Summary - Fo Generation Female Rats B12 Absolute Feed Consumption Values (g/day) Precohabitation - Summary - Fo Generation Female Rats ~~ B-13 Relative Feed Consumption Values (g/kg/day) Precohabitation - Summary - Fo Generation Female Rats ~~ B-14 Matemal Absolute Feed Consumption Values (g/day) - Gestation - Summary - Fo Generation Female Rats B15 Matemal Relative Feed Consumption Values (g/kg/day) - Gestation - Summary - Fo Generation Female Rats B16 Matemal Absolute Feed Consumption Values (g/day) - Lactation - Summary - Fo Generation Female Rats B17 Matemal Relative Feed Consumption Values (g/kg/day) - Lactation - Summary - Fo Generation Female Rats B-18 Natural Delivery Observations - Summary - Fo Generation Female Rats B19 Litter Observations (Naturally Delivered Pups) - Summary - F1 Generation Litters. B-20 Clinical Observations from Birth to Day 21 Postpartum - Summary - F1 Generation Pups 8-23 Necropsy Observations - Summary - F1 Generation Pups ~~ B-24 Clinical Observations - Individual Data - Fo Generation Female Rats B25 Necropsy Observations - Individual Data - Fo Generation Female Rats B28 vi 01007 SUBJECT PAGE Table 21. Body Weights- Precohabitation - Individual Data - Fo Generation Female Rats 830 Table 22. Matemal Body Weights - Presumed Gestation - Individual Data - Fo Generation Female Rats B-36 Table 23. FMoatGeemnaelraBtoidoyn FWeeimgahltesR-aLtasctation - Individual Data - B39 Table 24. Table 25. Feed Consumption Values - Precohabitation - Individual Data - Fo Generation Female Rats B-42 IMnadtievimdaulalFDeaetdaC-oFnosuGmepnetriaotnioVnalFueemsa-lPerReastusmed Gestation - B44 Table 26. DMaattaem-aFloFGeeenderCaotnisonumFpetmiaolneVRaaltuses - Lactation - Individual B47 Table 27. Natural Delivery, Implantation Sites, and Pup Viabilty Fa1ndGeSneexra-tIinodnivLidiutaelrsData -- Fo Generation Female Rats/ B49 Table 28. Pup Body Weight Litter Averages from Birth to Day 21 Postpartum - Individual Data - F1 Generation Litters B-51 Table 29. IPnudpiviBdoudaly DWaetiagh-tFs1fGreonmeBriarttihotno PDuapys21 Postpartum - 853 Table 30. Pup Vital Status and Sex from Birth to Day 21 Postpartum - Individual Data - F1 Generation Pups B-65 Table 31. CIlnidniivciadluaOlbsDeartvaa-tiFo1nsGefnreormatBiirotnh to Day Pups 21 Postpartum - B67 Table 32. Necropsy Observations - Individual Data - 1 Generation Pups B68 APPENDIX C- PROTOCOL AND AMENDMENT C1034 APPENDIX D - SDETVAINADTAIRODNSOFPREROAMTTIHNEG PPRROOTCOECDOULREASNDOFTHTEHE TESTING FACILITY D1 vi; 01003 SUBJECT APPENDIX E - TEMPERATURE AND RELATIVE HUMIDITY REPORTS APPENDIX F- STATEMENT OF THE STUDY DIRECTOR APPENDIX G - QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT PAGE Et0E9 F-1 G110G6 viii 010909 418-015:PAGE I-1 TITLE: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-015 SPONSOR'S STUDY NUMBER: T-6295.14 I. SUMMARY AND CONCLUSION A. Methods' Twenty-four presumed pregnant Cri:CDGBR VAF/PIus (Sprague-Dawley) rats were assigned to three dosage groups (Groups I through Ii), eight rats per dosage group. The rats were the vehicle, 0.5% Tween 80 administered the test article, PFOS (FC-85), in reverse osmosis membrane processed or deionized water(R.O. deionized water), orally (via gavage), once daily beginning 43 days prior to 0 (Vehicle), 0.1 cohabitation until confirmed evidenced of and 1.6 mg/kg/day were administered at a mating. dosage Dosages of volume of 5 mlikg. A.A. FoGeneration Rats: The female rats were observed for viability at least twice each day of the study. aTbhoertriaotnss,weprreemeaxtaumriendeedlifvoerricleisniacnaldodbesaetrhvsatbieofnosreofaenfdfeacptsproofxtihmeatteelsty aorntieclhe,our aafptpeeradroasnacgee.onRcaetsdawileyrdeuorbinsgeravlleodtfhoerrcpleinriicoadlsobofsesrtvuadtyi.onBsoadnydwegiengehrtaslwere recorded daily during the dosage and postdosage periods and at sacrifice. Feed gceosntsautmipontiaonndvoanludeasywse1r,e4,re7c,o1r0deadnwdee1k4loyf plraicotrattioocno(haDbLista1t,i4o,n,7,da1i0lyadnudri1n4g). `The female rats were evaluated for duration of gestation, litter sizes and pup viability at birth. Pups that either appeared stillbom or that died before initial Mexaatmeimnaaltiboenhaovfitohreolfitttehres dfoarmvsiabwialistyewvearleuaetxeadmdianileydwfhorenvittahlesptautpuss waterbierth. examined during the 21-day postpartum period. Observed matemal behavior was recorded on DLs 1,4, 7, 10, 14 and 21. a. Detailed descriptions of all procedures used in the conduct of this study are provided in the (PROTOCOL AND appropriate sections AMENDMENT). of this report and in APPENDIX C 01010 418-015:PAGE 1-2 Urine and fecal sample were collected from female rats for the following intervals: one day prior to initiation of cohabitation to the following morning, days 6107, 14 to 15 and 20 to 21 of presumed gestation (DGS 6 to 7, 14 to 15 and 2010 21), and DLs 21 to 22. Following each 24-hour collection interval, samples were shipped to the Sponsor for analysis. Blood samples were collected from each of the matemal rats on the day cohabitation was initiated (prior to cohabitation), DG 7, 15 and 21, and DLs 14 and 22. Serum samples were shipped to the Sponsor for analysis. All surviving rats assigned to the study were sacrificed on DL 22 following the final collection interval for urine and fecal samples and blood sample collection and a gross necropsy of the thoracic, abdominal and pelvic viscera was. performed. Tissues with gross lesions were retained. The number and distribution of implantation sites was recorded. A liver section from each dam was collected and shipped to the Sponsor for analysis. A2. F1Generation Litters: Day 1 of lactation (postpartum) was defined as the day of birth and was also the first day on which all pups in alitterwere individually weighed. The litters were observed for viability at least twice each day during the postpartum period. Litters were observed for clinical observations and general appearance once daily during the postpartum period. The pups in each litter were counted once daily. Body weights were recorded on DLs 1 (birth), 4, 7, 14 and 21 Pups found dead were examinedforgross lesions and for the cause of death. For all pups found dead on DLs 2 to 4, all lungs were preserved. On DL 4, litters were culled to five male pups and five female pups per litter, where possible. The lungs and the livers were collected from the first ten pups culled determined to be at DL 4 from each dosage group (irrespective of iter). The lungs and the livers were individually retained. Remaining culled pups were sacrificed and discarded without evaluation. All remaining pups on study were sacrificed and examined for gross lesions on DL 21. Gross lesions were retained. The liver from each pup was collected, pooled (per litter) and shipped to the Sponsor for analysis. Blood samples were collected and pooled (per liter). Serum samples were shipped to the Sponsor for analysis. 01011 B. Results 418-015:PAGE 13 No deaths or premature deliveries were attributed to PFOS treatment. One vehicle control group rat was injured and died after orbital sinus bleeding on DG 15. All other rats survived to scheduled sacrifice. All adverse clinical observations during the precohabitation, gestation and lactation periods were considered unrelated to the test article. A red substance was found in the thoracic cavity of the rat that died. All other rats appeared normal at necropsy. Rats administered the 1.6 mg/kg/day dosage of the test article had reduced body weight gains or body weight losses in each week of the precohabitation period. Dams in the 1.6 mg/kg/day dosage group had reduced body weight gains during the first week of the gestation period (DGs 0 to 7). Body weight gains were then increased as compared to the control group value on DGs 7 to 10, 13 to 15 and 1510 18. Reflecting this rebound effect, body weight gains were increased for the entire gestation period (DGs 0 to 20) in the 1.6 mg/kg/day dosage group, as. compared to the control group value. Bodyweight gains for dams administered the 1.6 mg/kg/day dosage of the test article during the precohabitation period generally continued to be increased during the lactation period. Absolute (g/day) and relative (g/kg/day) feed consumption values were reduced in the 1.6 mg/kg/day dosage group in each week of the precohabitation period. Dams in the 1.6 mg/kg/day dosage group had reduced absolute and relative feed consumption values during the first week of the gestation period (DGs 0107). Feed consumption values were then generally comparable to control group values for the remainder of the gestation period. Absolute and relative feed consumption values in this dosage group were slightly reduced, as compared to control group values, for the entire gestation period (DG 0 to 20). Absolute and relative feed consumption values during the lactation period were reduced in the groups administered 0.1 or 1.6 mg/kg/day of the test article during the precohabitation period, however the reductions were not strictly dosagedependent Administration of the test article at dosages as high as 1.6 mg/kg/day did not adversely affect any parameter evaluated at natural delivery or during the 22-day lactation period. No clinical or necropsy observations in the F1 generation pups were attributable to dosages of the test article as high as 1.6 mg/kg/day. 001012 C. Conclusion 418-015:PAGE 14 TpPhhFaeOmmSpauctrorpkeoiasntemeetoniftctsdhueorfsitPnugFdyOt,hSeaispnrseFt-oactogehedanbieinrtatathtiieoopnnroppterorecigoolnd,a.nwtTahfsiesmtoarleepevoarrlatutaisntfceolltulhdoeewsinsgtudy dIaabtoaraftroormiets,heInicn.-ffTehpeorpthiaornmwahciockhinweatsiccsoanmdpuclteesdthaattAwregrues cRoelsleecatrecdhduring the in-ife portion of the study were sent to the Sponsor for analysis and will be reported separately. It is the responsibility of the Sponsor, 3M Corporate Toxicology, to combine the pharmacokinetic report. in-life results with the analytical results into a final WAOst gos2 9 EMxieldcruetdivSe. DCihrreicsttoiarno,fPRhe.sD.s,arFcehllow, ATS Date 1 Alen Alan M. Hoberman, Ph.D., DABT Director of Research 23 Ju JP Date bo. 7 dA zy ow ARssomcoiantde GD.irYeocrtko,rForf-oRe.s.eDaArcBhTand Date Study Director 001013 418-015:PAGE II-1 I. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study: Ad. Sponsor: 3M Corporate Toxicololgy, 3M Center, Building 220-2-02, St. Paul, Minnesota 55144-1000 A2. Testing Facility: Argus Research Laboratories, Inc., 05 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 A3. Study Number: 418-015 Ad. Sponsor'sStudyNumber: T-6295.14 AS. Purposeof the Study: The purpose of this study was to evaluate the pharmacokinetics of PFOS in Fo generation and F1 generation rats during gestation and lactation following cessation of PFOS treatment of CH:CDBR VAF/Plus female rats at confirmed mating. AS. Study Design: The requirements of the U.S. Food and Drug Administration (FDA)" were used as the basis of study design. AT. Regulatory Compliance: The study was conducted in compliance with Good Laboratory Practice (GLP) regulations of the U.S. Food and Drug Administration (FDA)? the Japanese Ministry of Health and Welfare (MHW) and the European Economic Community (EECY. 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. 01013 418-015:PAGE Il-2 AB. Ownership of the Study: `The Sponsor owns the study. All raw data, analyses, reports and preserved tissues are the propertyofthe Sponsor. AS. Study Monitor: Marvin T. Case, D.V.M., Ph.D. A10. Alternate Study Monitor: Andrew M. Seacat, Ph.D. A1. Study Director: Raymond G. York, Ph.D., DABT (Associate Director of Research) A112. Technical Performance: John F. Bamett, B.S. (Directorof Laboratory Operations) Paul E. Ciotti (Team Leader) Todd J. Killino, B.S. (Laboratory Technician) A.13. ReportPreparation: Raymond G. York, Ph.D., DABT Jo Ann Frazee, M.S. (Study Coordinator) Denise P. Gasiorowski (Data Management Specialist) Karen G. Parker, A.A. (Report Administrator) A.14. ReportReview: Mildred S. Christian, Ph.D., Fellow, ATS (Executive Director of Research) Alan M. Hoberman, Ph.D, DABT (Director of Research) A.15. Date Protocol Signed: 16 November 1998 01015 418-015:PAGE I13 A16. Dates of Technical Performance: Rat Arrival Date Dosage Period [43 dayspriorto the first day of cohabitation* until confirmed mating (DG 0)] Cohabitation Period DG*0 Delivery Period (DL 1) Sacrifice of Fo generation rats not selected for continued evaluation DL 4 Culling (pups not selected for continued observation) DG 25 Sacrifice (dam with no confirmed date of mating) DL 21 Scheduled Sacrifice (F1 generation pups) DL 22 Scheduled Sacrifice (Fo generation dams) 17 NOV 98 23 NOV 98 - 08 JAN 99 04 JAN 99 PM - 09 JAN 99 AM 05 JAN 99 - 08 JAN 99 26 JAN 99 - 30 JAN 99 15 JAN 99 29 JAN 99 - 02 FEB 99 02 FEB 99 15 FEB 99 - 19 FEB 99 16 FEB 99 - 20 FEB 99 AAT. Records Maintained: `The original report, raw data and reserve samples of the test article and vehicle components are retained in the archives of Argus Research Laboratories, Inc. Any preserved tissues are retained in the archives of the Testing Facilty for one year after the mailing of the final report, after which time the Sponsor will decide their final disposition. All unused prepared formulations were discarded at the Testing Facility. All remaining bulk test article was retumed to the Study Monitor upon completion of all work with the test article. B. Test ArticleInformation: B.1. Description: PFOS (FC-95) - off-white powder B.2. Lot/Batch Number: 217 (Expiration date: May 2000) a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), item 1. b. DG is used as an abbreviationfor day of (presumed) gestation. c. DLis used as an abbreviation for day of lactation or day postpartum. 0101 418-015:PAGE 114 B.3. Date Received and Storage Conditions: The test article was received on 21 October 1998, and stored at room temperature. B.4. SpecialHandlingInstructions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles or safety glasses and a face-shield) were taken when handiing the bulk test article and prepared suspensions. B.S. AnalysisofActivity: Information regarding the identity, composition, strength and purity of the test article is on file with the Sponsor. C. Vehicle information: C1. Description: 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Tween 80 - a clear or yellow viscous liquid C.2. LotNumbers: Tween 80 - M29477 and MO3HOS C.3. Dates Received, Source and Storage Conditions: Tween 80 was received from J.T. Baker, Philipsburg, New Jersey, on 17 September 1998 (lot M29477) and 3 December 1998 (lot MO3HOS), and stored at room temperature. The R.O. deionized water is available from a continuous source at the Testing Facility and is maintained at room temperature. C4. Special Handling Instructions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles or safety glasses and a face-shield) were taken when handling the vehicle. co1017 418-015:PAGE [1-5 C5. Analysis of Purity: Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. D. Test Article Preparation and Storage Conditions: Suspensions of PFOS (PC-95) were prepared daily at concentrations of 0, 0.02, and 0.32 mg/mL. Prepared formulations were stored at room temperature. D.1. Sample Information: 5Sample Type Concentration al ORaettasined | CStoonrdaigtei.ons Shipped To__| we | [2NEoovess [rom Joe [BRDate Shipped 2NoVSs BulkTestAvice Room Testing VerzdizComponentsi oo [2 ReTswewerveeno80 lam | novee| mam RO.LoWoatseHros | sSmmL || 1109nDoEvCsSSs | temperate [ER Joo | woole | FaCHY ome 22773a3N9m99 3 Dn uplica atetshael mprelmeasis wneinrgeaamkpelnesowmetrherefrisntean 85asbapcrkeuppasr.atBiaonc.kOunp essaammpsleowfereaSchesdetwazsesnh(i7p0Cfoorr elon)andwilbe scared a th Testing Faciyupon ierequestof te Sponsor D2. AnalyticalResults: Information on the stability and homogeneity of the prepared formulations and of the stability of the bulk test article are on file with the Sponsor. Data verifying the stabilityofthe test article in the vehicle for 48 hours under the conditions of administration are on file with the Sponsor. Records were maintained to documenthow the test article formulations were prepared. Results of the concentration analyses were not available at the time of the writing of this report E. TestSystem: EA. Species: Rat 03i013 418-015:PAGE Il-6 E.2. Strain: Cri:CDBR VAF/Plus (Sprague-Dawley) E3. Supplier (Source): Charles River Laboratories, Inc., Raleigh, North Carolina Ed. Sex: Female (Note: Male rats were used only for the purposes of breeding and are not considered part of the Test System.) E5. Rationale for Test System: The Crl:CDBR VAF/Plus (Sprague-Dawley) rat was selected as the Test System because: 1) this strain of rat was used in the reproductive and developmental toxicity studies; 2) historical data and experience exist at the Testing Facility'"; and 3) the test article is pharmacologically active in the species and strain. ES. 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 37 14 SEP 98 65 days 181-222 192-231 E7. 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 ES. Method of Randomization: 112 13 JAN 98 78 days 300 - 356 515-893 Upon arrival, rats were assigned to individual housing on the basis of computergenerated random units. After acclimation, 36 virgin female rats were placed into groups on the basis of physical appearance and body weights recorded during acclimation. Female rats were assigned to three dosage groups (Groups I through Ill), 12 rats per dosage group, using a computer-generated (weight-ordered) randomization procedure. After these 36 rats were cohabitated, 041019 418-015:PAGE II-7 sight mated female rats (those with confirmed evidence of mating) were selected for the study from each dosage group. On DL 4, a table of random units was used to cull the litters to five male pups and five female pups per litter, where possible. ES. System of Identification: E.9.a. Fo Generation Rats: 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 before cohabitation. Each rat was individually identified with a Monel self-piercing ear tag (Gey Band and Tag Co., Inc., No. MSPT 20101). E.9.b. F1 Generation Pups: Pups were not individually identified during lactation; all parameters were evaluated in terms of the liter. F. Husbandry: FA. Research Facility Registration: USDA Registration No. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 ot seq. F2. Study Rooms: The study rooms were maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum of ten changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters. Room temperature and humidity were monitored constantly throughout the study. Room temperature was targeted at 64F to 79F (18C to 26C); relative humidity was targeted at 30% to 70%. See APPENDIX E (TEMPERATURE AND RELATIVE HUMIDITY REPORTS). F.3. Housing: All cage sizes and housing conditions were in compliance with the Guide for the Care and Use of Laboratory Animals. Fo generation rats were individually housed in stainless steel, wire-bottomed cages, except during the cohabitation and postpartum periods. During cohabitation, each pair of rats was housed in the male rat's cage. Beginning no later than DG 20, Fo generation female rats 01020 418-015:PAGE 11-8 were urine individually housed and fecal samples. in nesting boxes, except during collection intervals for During these collection intervals, the female rats were housed individually in metabolism cages. Each dam and delivered litter were housed in a common nesting box during the postpartum period. Fa. Lighting: An automatically-controlled fluorescent light cycle was maintained at 12-hours light:12-hours dark, with each dark period beginning at 1900 hours EST. F5. Sanitization: Cage pan liners were changed approximately three times each week. Cages were changed approximately every other week. F.6. Feed: Rats were given ad libitum access to Certified Rodent Diet #5002 (PMI Nutrition Intemational, St. Louis, Missouri) in individual feeders. F.7. FeedAnalysis: Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to with the results of this have study. been present in the feed that would have interfered FB. Water: Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad ibitum from an automatic watering access system and/or individual water bottles. 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. cic 418-015:PAGE 11-9 Neither the Sponsor nor the Study Director nor the Sponsor was aware of any potential contaminants likely to have been present in the water that would have interfered with the results of this study. F.10. Bedding: Bed-0'cobs was used as the nesting material (The Andersons' Industrial Products Groups, Maumee, Ohio). F.11. Bedding Analysis: Bedding was changed as often as necessary to keep the animals dry and clean. Analyses for possible contamination were conducted annually and documented in the raw data. Neither the Study Director nor the Sponsor was aware of any agent present in the bedding that was known to interfere with the resultsofthis study. G. Methods: G.1. DosageAdministration: osage| Number Group | (mohgidayy| (mgmt) | (ming) |" Rats AT37s26s- iTR3a7g2t8N,numTe3b7e3dr1s. [oom[0[oe Te ERE [oTon J om [0To["og137n48-g1374i9 TMe| reTee [ov |womens| a. The test aricte was considered 100% pure forthe purpose of dosage calculations. G2. Rationale for Dosage Selection: Dosages were selected on the basisof a previous study conducted with the test article (Argus Research Laboratories, Inc. Protocol 418-008). In this study the Fo generation maternal and paternal no-observable-effect-level (NOEL) of PFOS was 0.1 mg/kg/day (0.4 mg/kg/day and higher dosages caused reductions in body weight gain and reduced feed consumption values). The Fo generation reproductive NOEL was greater than 3.2 mg/kg/day; no effects on mating, fertility or estrous cycling occurred. The NOEL for viability and growth in the F'1 generation offspring was 0.4 mg/kg/day (1.6 mg/kg/day and higher dosages viability, growth acnadusseudrvipvraeli)m.plantation loss and reductions in litter size, pup 01022 418-015:PAGE 11-10 The F1 generation matemal and patemal NOEL of PFOS was 0.1 mg/kg/day (0.4 mg/kg/day dosage caused reductions in body weight gain and reduced feed consumption values). The F1 generation reproductive NOEL was greater than a dosage of 0.4 mg/kg/day; no effects on mating or fertility occurred. The NOEL for viability and growth in the F2 generation offspring was 0.1 mg/kg/day (0.4 mg/kglday dosage caused stillbirths and reductions in litter size, pup viability, growth and survival). G.3. RAoudtmeionfistration: Oral (gavage) G.4. Rationale for Routeof Administration: The oral (gavage) route was selected for use because: 1) this was the route of administration in the developmental and reproductive toxicology studies; and 2) it is one of the possible routes of human exposure. G5. Freque f istration: G.5.a. Fo Generation Female Rats: Appropriate dosages of the test article or vehicle were administered orally (via gavage) once daily to female rats beginning 43 days prior to cohabitation until DG 0 (confirmed evidenced of mating, such as observation of spermatozoa in a `smear of the vaginal contentsor a copulatory plug in situ). Female rats were not given the test article or vehicle on DG 0. Dosages were 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.b. F1GenerationPups: F1 generation pups were not directly given the test article, but may have been possibly exposed to the test article during matemal gestation (in utero exposure) or via matemal milk during the lactation period. G6. Length of Study: Approximately 14 weeks a. See APPENDIX D, item 2. b. See APPENDIX D, item 1. 61023 418-015:PAGE Il-11 G.7. Methodof Study Performance: G.7.a. Fo Generation Rats: After acclimation and 43 days of dosage administration, 36 healthy virgin female rats were placed into cohabitation with 36 breeder male rats (one male rat per female rat maximum in of the five male days. rat's cage). The cohabitation Mating was evaluated daily period during consisted ofa the cohabitation period. Female rats with contents or a copulatory spermatozoa observed in a plug in situ were considered smear of the vaginal to be at DG 0 and retumed to individual housing. were selected for the Twenty-four mated study, as previously female rats, discussed. eight per dosage group, The female rats were observed for viability at least twice each day of the study and for general appearance at least once during acclimation. `examined for clinical observations of effects of the test article, The rats were abortions, also premature deliveries and deaths before and approximately one hour after dosage. Rats were observed for clinical observations and general appearance once daily during all other periods of study. rBeocdoyrdweedigdhatilsywdeurreinrgetchoerddeodsoangceeadnudripnogstacdcolsiamgateiopne.rioBdosdaynwdeiatghstascriwfeircee. Feed consumption cohabitation, values were daily during recorded gestation once during acclimation, weekly prior and on DLs1,4, 7, 10 and 14. Feed to consumption was not tabulated after DL would begin to consume matemal feed. 14, when it was expected that pups The female rats were evaluated for duration of gestation (DG 0 to the day the f(ilrisvte pbuopmwpausposbsoenlryv)eda)n,dlpitutepr sviiazbeisli(tydeaftinbierdtha.sPalulppsutphsatdeeiltihveerread)p,peliavreeldittsetrislilzboern or that died vital status before at birth. initial The examination of the litters lungs were removed and for viability immersed were examined for in water. Pups with lungs that sank were considered stillborn; pups with lungs that floated were dcoanmssidwearsedevliavleubaotrendadnadiltyowhhaevne tdhieedpushposrtwleyraefteerxabimrithn.edMadutreimnaglthbeehavior of the 42.17-,da1y0,po1s4tapnardt2u1m.peDreivoida.tiOobnssefrrvoemdemxapetcetmeadlmbaethearvniaolrbweahsavrieocrowrederde ornecDorLdsed1,, if and when present, on all other days during the postpartum period. G.7.b. F1 Generation Rats: Day 1 of lactation (postpartum) was defined as the day of birth and was also the first day on which all pups in were recorded after all pups a in litter were individually alitterwere delivered weighed (pup and groomed body weights by the dam). 01024 418-015:PAGE I-12 The litters were observed for viability at least twice each day during the postpartum period. Dead pups observed at these times were removed from the nesting box. Litters were observed for clinical observations and general appearance once daily during the postpartum period. The pups in each litter were counted once daily. Bodyweights were recorded on DLs 1 (birth), 4, 7, 14 and 21. G8. Pharmacokinetic Sample Collection: G.8.a. Fo Generation Rats: Urine and fecal sample were collected from female rats for the following intervals: one day prior to initiation of cohabitation to the following morning, DGs 6107, 14 to 15 and 20 to 21, and DLs 21 to 22. Following each 24-hour collection interval, samples were collected into centrifuge tubes, placed on dry ice and stored frozen (-70C or below) and shipped, frozen on dry ice, to the Sponsor for analysis. Blood samples were collected from each of the matemal rats following removal from metabolism caging (prior to test article or vehicle administration) on each of the following days: on the day cohabitationis initiated (prior to cohabitation), DGs 7, 15 and 21, and DLs 14 and 22. On all days of collection except DL 22, blood samples (approximately 1 mL each) were collected from the orbital sinus. On DL 22, blood samples (approximately 4 mL each) were collected via the inferior vena cava. Blood was collected and transferred into serum separator tubes. The samples were spun in a refrigerated centrifuge. The serum was transferred into polypropylene tubes labeled with the. study number, rat identification, date of collection, study day and collection timepoint. All samples were immediately frozen on dry ice and maintained frozen (-70C or below), and shipped, frozen on dry ice, to the Sponsor for analysis. G.8.b. F1 Generation Litters: On DL 21, blood samples were collected from all remaining pups on study. Blood samples were collected via the inferior vena cava from each pup, pooled (per litter) and transferred into serum separator tubes. The samples were spun in a refrigerated centrifuge. The serum was transferred into polypropylene tubes labeled with the study number, rat identification, date of collection, study day and collection timepoint. All samples were immediately frozen on dry ice and `maintained frozen (-70C or below) and shipped to the Sponsor for analysis. 01025 418-015:PAGE 11-13 G9. Gross Necropsy: G.9.2. Fo Generation Rats: Female rats, with or without confirmed evidence of mating, that were cohabited but not assigned to the study, were sacrificed by carbon dioxide asphyxiation and discarded without further evaluation. All surviving rats assigned to the study were sacrificed by carbon dioxide asphyxiation on DL 22 following the final collection interval for urine and fecal samples and blood sample collection. A gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Tissues with gross lesions were preserved in neutral buffered 10% formalin for possible future evaluation. Representative photographs of matemal lesions are available in the raw data. The number and distribution of implantation sites was recorded. A liver section (fight lateral lobe) from each dam was collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. All other maternal tissues were discarded. The rat that did not delivera litter was sacrificed on DG 25 and examined for gross lesions. teri were stained with 10% ammonium sulfide to confirm the absence of implantation sites. The rat that was found dead was examined for the causeofdeath on the day of the death. The rat was examinedforgross lesions. A liver section (right lateral lobe) was collected and stored, as described above. Pregnancy status was recorded; fetuses were examined to the extent possible. G.9.b. F1GenerationPups: All pups culled on DL 4 were sacrificed via decapitation. The lungs and the livers were collected from the first ten pups culled determined to be at DL 4 from each dosage group (irrespective of litter) and preserved for possible future histopathological evaluation. The lungs were individually retained in Bouin's solution, and the livers were individually retained in neutral buffered 10% formalin for possible future evaluation. Remaining culled pups were sacrificed and discarded without evaluation. Pups found dead were examined for gross lesions and for the cause of death. For all pups found dead on DLs 2 to 4, all lungs were preserved in Bouin's solution for possible future evaluation. All remaining pups on study were sacrificed on DL 21 by carbon dioxide `asphyxiation and examined for gross lesions. Gross lesions were preserved in neutral buffered 10% formalin. The liver from each pup was collected, pooled 01026 418-015:PAGE II-14 (per litter), frozen and stored (-70C or below) and shipped to the Sponsorfor analysis. G.10. Statistical Analyses: Averages and percentages were calculated.Litter values were used where appropriate. 01027 mW. RESULTS 418-015:PAGE Iil-1 A. Mortality, Clinical and Necropsy Observations (Summaries - Tables 1 and 2; Individual Data - Tables 19 and 20) AA. Mortality No deaths or premature deliveries were attributed to PFOS treatment. One vehicle control group rat was injured and died after orbital sinus bleeding on DG "15. All other rats survived to scheduled sacrifice. A2. Clinical Observations All adverse clinical observations during the precohabitation, gestation and lactation periods were considered unrelated to the test article because the incidences were not dosage-dependent and/or the observation occurred in only one rat in a group. These observations included dental problems (missing, broken and/or misaligned incisors), chromodacryorrhea, abrasion on the forepaw, localized alopecia on the head or limbs, chromorhinorrhea, exophthalmos, hemorrhagic area on the eye, urine-stained abdominal fur, missing eye, comeal opacity, swollen area on chestoraround eye, red, dried perioral substance, tom ear and soft or liquid feces. A red perinasal substance, blue paws and gasping were agonal signs for the rat that died. A3. Necropsy Observations A red substance was found in the thoracic cavity of the rat that died. All other rats appeared normal at necropsy. B. Body Weights and Body Weight Changes (Figure 1; Summaries Tables 3 through 8; Individual Data - Tables 21 through 23) B.1. Precohabitation Rats administered 1.6 mg/kg/day dosage of the test article had reduced body weight gains or body weight losses in each week of the precohabitation period. Reflecting these effects of the test article, body weight gains were 46.1% of the control group value for the entire precohabitation period (DSs 1 to 43) in the 1.6 mg/kg/day dosage group. Precohabitation body weights and body weight gains were unaffected by the 0.1 mg/kg/day dosage of the test article. 01023 B2. Gestation 418-015:PAGE ll2 Dams administered 1.6 mg/kg/day dosage of the test article during the precohabitation period (treatment ended on DG 0) had reduced body weight gains during the first week of the gestation period (DGs 0 to 7). Body weight gains were then increased as compared to the control group value on DGs 7 to w10e,re13intcor1e5asaenddfo1r5thtoe 1e8n.tirReefgleescttaitnigonthpiesrrieodbo(uDnGdsef0fetcot,20b)odinytwheeig1.h6t mggai/nksg/day dosage group, as compared to the control group value. Gestation body weights and body weight gains were unaffected by the 0.1 mg/kg/day dosage of the test article. B3. Lactation Barotidcylewdeuirgihntggtahiensprfeocrodhaabmistaatdimoinnpiesrtieordedgetnheera1l.l6ymcgo/nktgi/ndueady tdoosbaegiencorfetahseedtest during the lactation period. Lactation body weights and body weight gains were unaffected by the 0.1 mg/kg/day dosage of the test article. C. A(bSsuomlmuatrei(egs/d-aTy)abalensd 9Retlhartoiuvegh(g1/4k;g/Idnadyi)viFdueaeldDCaotnas-uTmapbtlieosn2V4alues through 26) CA. Precohabitatior Arbesdoulcuetde i(ngt/hdeay1).a6 nmdg/rkelga/tdiavey (dgo/skag/gdeayg)rfoeuepdincoenascuhmpwteieokn ovfatlhueesprweecroehabitation period. reduced Reflecting these effects of the for the entire precohabitation test article, period (DSS feed 1 to consumption 43) in the 1.6 values were mg/kg/day dosage group, Precohabitation feed consumption dosage of the test article. values were unaffected by the 0.1 mg/kg/day C2. Gestation Dams administered 1.6 mg/kg/day dosage of the test article during the prerleactoihvaebfieteadticoonnpseurmipotdi(otnrevaatlmueenstdeunrdinegdtohne DfirGst0w)eheakdorfetdhuecgeedstaabtsioolnutpeerainodd (coDnGtrsolOtgoro7u)p. vFaeleudescfoonrstuhmeptrieomnaivnadleureosfwtehreegteshteantgioenneprearliloyd.coAmbpsaorlaubtleeatnod 01029 418-015:PAGE Ill-3 relative feed consumption values in this dosage group were slightly reduced, as compared to control group values, for the entire gestation period (DG 0 to 20). Gestation feed consumption values were unaffected by the 0.1 mg/kg/day dosage of the test article. C3. Lactation Absolute and relative feed consumption values during the lactation period were reduced in the groups administered 0.1 or 1.6 mg/kgiday of the test article during the precohabitation period, however the reductions were not strictly dosagedependent. D. Natural Delivery and LitterObservations (Summaries - Tables 15 and 16; Individual Data - Tables 27 through 30) Natural delivery observations were based on 7, 8 and 7 pregnant rats in each of the three respective dosage groups. Administration of the test article at dosages as high as 1.6 mg/kg/day did not adversely affect any parameter evaluated at natural deliveryor during the 22-day lactation period (duration of gestation, averages for implantation and live litter sizes, numbers of dams with no livebom pups or all pups dying during lactation, gestation, viability and lactation indices, surviving pups per litter, litter size at `weighing, pup weight per liter and pup sex ratios). E ical m Birthto Day 21 P Necropsy Observations (Summaries - Tables 17 and 18; individual Data - Tables 31 and 32) No clinical or necropsy observations in the F1 generation pups were attributable to matemal dosages of the test article as high as 1.6 mg/kg/day because: 1) the incidences were not dosage-dependent; and/or 2) the observation occurred in only one or two pups. The only adverse clinical observation was a black tip of tail in one 0.1 mg/kg/day dosage group pup. No milk in stomach occurred in 0, 1 and 2 pups that were found dead in the three respective dosage groups. All pups appeared normal at necropsy on DL 4 or 21 21030 418-015:PAGE Ili REFERENCE: 1. U.S. Food and Drug Administration (1994). Intemational Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Federal Register, September 22, 1994, Vol. 59, No. 183. 2. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. 3. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice StanfdoraSafretdy Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 4, European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratorypractice. Official Jounal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. 5. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 6. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin. Psychiat. 45(3):7-10. 7. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River Crt:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 8. Institute of Laboratory Animal Resources (1996). Guideforthe Care and UseofLaboratory Animals. National Academy Press, Washington, D.C. 9. Salewski, E. (1964). Frbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. 01031 APPENDIAX REPORT FIGURE 01032 BODY WEIGHTS Fo GENERATFIiOgNurF1eEMALE RATS wl : | o ol wy7 ER.E - 3 i - RL ry gw 2" Lo PE = FT | |= [1 3 - Hams Spade ee em EG ER 3 e oavorstuor oAY oFGesTATION OAYoF LicTATION 8 Ep SSm ERs E, | I = APPENDIX B REPORT TABLES 021034 Epo ci I essen TLSL LD US ) BRET Thoraumeptec1Scoioarton. B<RrlchSSeaEbrerEants TEorSely cooutisartiscle tn" r,m tone rs ses . g2 2 ; S & 2 hrm I A vt --e wn om rn --. Er ll pupa ous ewe sv & Sh Shia te GrtyTE iSen nS tary S'S hrougn of ser 8 2g88 82 3 Dam 13731 had un accidental death on day 15 of gestation. 2 commesarons woe wa un xe onenosirevn, ssn [A ------ ---- Wawa we we wa owe wa we we L TL CI Shara oeL Ti Tekh re ood wen 3 Spay on iy Cht-- o To serGroup 11 rate vere dosed -3 332 28 @&  ISS LTT s Mr on day 3 0 po"of peavensdain fo Co ER AI BS E A add - i : : | FhEES ETE sSp A SALRre eecos t & 3 3 i83s m z | PROTOCOL 410-015, CANL (GAVAGE) PAMKACORINETIC RECOVERY STUDY OF F705 IN RATS (SKOMSOR'S STUDY WAGER, T-6295.14) Leh ST rr -- ali tne rotpLa ath Cre aces en's Sybase on eT ran sy 1 ree ee doe gg 23 38 s3 & ssnEn dT TE TTT 3 2 v en BL SLL he cco tn cnt actcte rapustion, aroun 1 rat sere doses 2 RB Fetus erore chs Cat ett okeefco Se mcioeetmd noh soapy | I of ary ; I os rn mm Je sms . EEERR a g ~ : 3 3 & 8 3 wenn wens. een os was ve earsn ;FE EL EE R 3 3: 22 ol in Sma. men ne & B3 EEELIE Lgi el ss pie : : &E 3 o EBTERRAer aSecon ht Fors ave dove wet a/hpron nsaays nsecEamo CrP 11 1s were does 8 rr rin ima main ano IT en : & g83 BE o EE EEECEhe ElEEEALTe !RIRUTD SILIy E SUSceie anTtcrSe prepucion, com 10 ee &33 zz EE. + wn Es oN rs BolEh el 0 Sa r hd EySETILL ee. g i 32 ? N 2 Co me es wpa war ar oa wens 0. arr maga alsa oe sors 3 EPRI SLI S10 SLT SL wn sect proton, ro 1 1 ar dns EwLith 67128 mafig/dey and Growp 111 ath vere dosed vith 3 my/nadey oa dave 1 Chiou 4 of seedy. & 2 3832 @ 55 eo wn ae - 2 D TE ETT tout 0 SER A WOS U stTmov wm oe & 2 & 2= > i = Eri toon TF 2 aanor asus avemsaeo co - oo BT ST I SE mdY a Su 13IEoc)h 0Se8r cE od eI n Sahruanyin taarteasthhresao ohey 1 TS re ened :. EH TELSe enh Sl g .2 & 2 2 a 33 Fo - PUY fa ;E E TntetE 0Se3 tISoLeT ts sTpi, 1 a sv 2 3 # 22 52 J. amr pr wise hE iSETn ST Sa I BAinL tent ariT el , ion. arene 11 ador.es PE ee Icnn mss wots ssp 32 s I5 a&222 3z rr mse. Tp. ness masa EE dT Foy ut Ee od SEety a Sty i 1 hls arinan daresSnofe ses u? 1 1 vere doses 3 #3 88 : pa bi wi "3 BE LE EE LR a nie na i TT me ves 338~ ap 22 3 &8 Fo min 3 # 3 T + ERE A E yn wy SSLe eee &3 882 2 FEEL : a ss sstomns emer hi. ets A.8. 2 oF ERE tn SS son race sts, ts re 3 BBR AILMatIa bIear sloItLIpop pac LicAes, InTcioesintTatetars iseomnane memvrivviiveing ppeuepae + oF > ag g2 3 EeyT nr tsotrn oi, 3 o 3 ACh TS eparny and Group 11 Tach ware dosed HED me mH/BraSy oh Seve') Chron 3 obsoniye? 1 TH re dosed 2 8 8a R NL TOTAL PRRGURMCY (GATS x SORE JLITTERGWITH omsenvATIONS or Tr o Bho narr retac aSctata yeuTilateihon: Ssehic sesEbeSoe'sStare aTRAE E, en vee on : 8 - a 22 az 4 8 &g 2 2 & : ;g 25 RanETBis TsT veem RE 3 = : &g 3 ose ewan menses, wssne/mons cr EE rer dt nin i - 3g 22 z8 g& ~ Be cui ce 1 rapt mtn int hs g 3 = Ire 8 88 i al Temeited tn an eFFOr In test article preparation, ats were dosed with 3 2 8 Er, oh JO HETR mrs or i IbE ie aavrros aen eeteie prorarae erwateroieta vith g a23 2 3#22 Q 3 a3 33 B 23 a@ 8 3 & 3 3 8 & &7 @ 2 i dl Ee EMIT ELEEIETLELEY MD a ---- 2 3 & 23 &  RTE ELIAmn wes no bo si pt 954 wi dt : g & TL er ron roteeneo re 1 ih ton, ts re de in a 5 ? S2o #2 ey EL = EEE SIE, i conics 1 nor co sce potion, sr oe in & 2 8 3 a3 & OE 3 2 o g 8 SL & DEERLEEESmap mnee meg 33 4 Cr omarcrow 111 iH oosack. 1.6wana a co : i v HEE ip wor PRR (VAUORSEXCLUDED ROW AVERAGES) ce - g goEn I EE EESE o E 1 ch rete a ebeens iecmt prion, ts ee dnd ih 3 2 2 23&? 25 8 E EETRE 0 cnt te bn cts mt, i z 2 3 2? LT0 tn cnt re te etn 28 s83 5a3 RT g3 o BE - - Eroeetrpreioraedenoehn Selalatioant this value ) & $g i FE ER ar mr 2 35g2$ :2R Li EE i o PEREFn iroevor i s f 3 FT HR Wiss wi es Ers tn 2ig DE SETTERERE0R7I.E RvEats oc es 4tm weremteemattoenppt, Erics wTt sStt 23 g3 i v Y. 8 Wp Wor AEOUANT (VALDES EXCLUOED PRON AVERAGES) or g Tham E8 ee ee & i E TETERAa NEE ma 8828g 0 0L-Ltn. es we det wn & 32 BRETlv mr A TL ALbc,. ses oe sv in & g2 &38 2 & Ay i E Ele aee TE "Mariana coinchoos 11 Law bomat Ce ears - A ers einen A ER Trae preparation, race vere dosed wich g 2$2 :2i g 3 EE & 5 2$ 8 Son Fo I HEE ti R g gg 8: &82 g Xe a TR Ia ate are doses wien ; :: :: 2 g :3 2 Be ET wm sor sn & 5 8 5 | alesTae Tone in vie grissiin, itn tn shih 5 Z :s & | ee . 8 3 i @ 8 ee? 2 ertsiriin beens 2 rss dlne 8 2 Tl TE TT Stee esecion, roe er oes win : & 83 gS3 q&@ 8 :i i: F-- J - I . BE EeHO CBR AREER ein . & g2 82 8 8 TeET 2 2 i. sn oe si g g g: & :8 ERE re ee ar . :g g a 88 5 2 g 2 & pb g Ee mm rt tl A AR ] FREE EEE EEE : : . g3 & 2 g 2 2 & 38 & i 8 8 | : ES amin a ehTe, 1 ce ere doves i: g ii3 ] oe 1 tm 1. sorscoins mv on co mE TT iu SRS 2 i & cd SEREEREE g 2 3 5 + Fh REMEi Te 5 s & g $ a 8 APPENDIX C PROTOCOL AND AMENDMENT 001105 418-015:PAGE C-1 rv, PRIMED!ICA - Argus30R5esSehaarecWmhayrLeBahbraoemr.a,toBPruAiteas1i.9nI0gn4cA.4 TolaToptheornaex:: ((321155))44433--88578107 PROTOCOL 418-015 SPONSOR'S STUDY NUMBER: T-6295.14 STUDY TITLE: Oral (Gavage) Pharmacokinetic Recovery Study of PFOS in Rats PURPOSE: pThhaermpaucropkoisneetoifctshoisfsPtFudOySisitnoFeovagleunaetreattihoen and cFe1ssgaetnieornatoifonPFraOtSs dturreiantgmegnetstoaftiCorn:aCnDdBlBacRtaVtAioFn/fPolluloswing female rats at confirmed mating TESTINGFACILITY: A9r0g5uSshReeesheyarDcrihveL,abBourialtdoirnigeAs, Inc. Horsham, Pennsylvania 19044-1287 Telephone: (215) 43-8710 Telefax: (215) 443-8567 STUDYDIRECTOR: ~~ Raymond G. York, Associate Director Ph.D., DABT of Research `SPONSOR: STUDY MONITOR: 33MM CCoenrtpeorr,atBeuiTlodxiincgo2l2o0g-y2-02 St. Paul, Minnesota 55144-1000 MTaerlveipnhoTn.eC:ase(6,51D)V.7M3.3,-5P1h8.D0. Telefax: (651) 733-1773 ALTERNATE STUDYMONITOR: TAenldeprheoMnwe.: Se(a6c5a1t), 5P7h5.-D.3161 Telefax: (651) 733-1773 001106 418-015:PAGE C-2 Protocol 41P8a-g0e152 REGULATORY CITATIONS: U.S. Food Guideline oannddetDercutgioAndmoifntiosxtircaittyiotno r(1e9p8r4o)d.uctIinotnemfaotrimoendailcCinoanlfeprreodnuccetso.n HFaerdmeornailsation; Register, September22, 1994, Vol. 59, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. fJoarpSaafneetsye MSitnuidsitersyoofnHDeraulgtsh, aMnHd WWelOfradriena(n1c9e87N).umGboeord21L,abMoarractohr2y6,Pr1ac9t9i7c.e Standard E`aucrcoeppetaannceEcboyntohmeicEuCroompmeuanniEtcyo(n1o9m88i)c. CoComumnucniiltdyecoifsainonOoEnC2D8 dJuelcyis1i9o8n9/roencotmh-e mendation on the European Ccoommmpulniiatniceesw:itLhepgriisnlactiipolne.sof32go(Nood. laboratory L 315; 28 practice. October): Official 1-17. Joumal of uY NC] This study will be conducted in compliance with the Good Laboratory Practice (GLP) regulations cited above. All changes Director and or revisionsofths protocol shall be the Sponsor, dated and maintained documented, signed with the protocol. by the Study Tanhde wQiulalliintsypAecstsucrriatinccalepUhnaitse(sQAofU)thweilsltauuddyititnhaeccporortdoacnolc,etwhiethrathwedSattaanadnadrdthOeperreapotritn,g Procedures of Argus Research Laboratories, Inc. TachceurfaitnaellyrerpeofrltecwtislltihneclruadwedaatsataotbetmaeinntedsidgunreidngbtyhtehpeeSrtfuodrymaDnicreeocftotrhtehastttuhdeyraenpdortthat salilgnaipfpilciacnatbdleeviGaLtPionrsegfurloamtiGonLsPwerergeulfaotliloonwsedocicnutrh,eecaocnhduwicltl of be the study. described Should in detail, together with how the deviation might affect the quality of integrity of the study. `SCHEMATICOF STUDYDESIGNANDSTUDYSCHEDULE: `See ATTACHMENT 1 to the protocol. 001107 418-015:PAGE C-3 Protocol 4P1a8g0e1s5 TAESTRTIANCDVELHICE LE: Identification: Test Article: NPhaymsei:cal Description: Lot/Batch Number: `Specific Gravity: Purity: Expiration Date: LPiFghOtS-c(olSoyrneodnpyomw:deFrC-.85). 217. ~0.6. 98.9%. May 2000. Information on the identity, composition, strength and purityofthe test article is on file `with the Sponsor. Vehicle: 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Supplier and lot identificationof Tween 80 to be documented in the raw data. NtoeibteheprrethseenStpionntshoervneohirctlheetShattudwyouDlidreicnttoerrfies raewwairtheotfheanreysuplottsenotfitahliscosnttuadym.inTahnetrsefliokreel,y no analyses other than those mentioned in this protocol will be. `conducted. SafetyPrecautions: Gloves, mask, appropriate eye protection and a uniform/lab coat are to be wom during formulation preparation and administration. The Material Safety Data Sheet (MSDS) is attached to the protocol (ATTACHMENT 2). Storage: Bulk Test Article: Vehicle Components: Prepared Vehicle: Prepared Formulations: Room temperature. Room temperature. Room temperature. Room temperature. telephone number. All test article shipments to the Testing Facility should be addressed to the attention of Julian Gulbinski, Man ofa Forg mulae tior ns, at the previously cited address and 001108 Shipments should cartons should be include labeled iapnpfroorpmraitaitoenlyc.onTcehrenirnecgipsiteonrtasgheocuolnddibteionnostiafinedd shipping in advance of shipment. 418-015:PAGE C4 Protocol 41P8a.g0e1s5 FORMULATION: Freqouf Perepnaractioyn: taFhdoemrimsnutilasbatitlriiatotyniosofn(tashrueesptoeensntsfiialoretniwsci)ltewhiiltnlhtebheeSpvpoernhesipocarlre.edforda4i8lyahotutrhseuTnedsetrintgheFaccoinlidtiyt.ioDnastoafverifying Detailed preparation procedures are attached to this protocol (ATTACHMENT 3). `AdjusftormPuerintyt: The test article will be considered 100%purefor the purpose of dosage calculations. TestingFacility ReserveSamples: The Sponsor will reserve a sample (1 g)of each lotof the bulk test article used during the course of this study. TheTesting Facility will reserve a sample (5 mL) of each lot of the vehicle components used during the courseofthis study. Samples will be stored under the previously cited conditions. ANALYSES: `Samples additional to those described below may be taken if deemed necessary during the course of the study. BulkTestArticle Sampling: No analyses of the bulk test artcie wil be conducted during the course of this study. Information on the stability of the bulk test article is on file with the Sponsor. Analyses of Prepared Formulations: Homogeneity and stabilityof prepared formulations is on file with the Sponsor. However, records will be maintained to document how the test article formulations were prepared. ConceofnTesttArrticale Ftormiulaotionns: Concentration of the prepared formulations will be verified during the course of this study. Duplicate samples (2 mL each) will be taken from the first and last preparation on the day prepared. One remaining samples will be sampleof retained at each set will be shipped for analysis; the the Testing Facility as backup samples. Backup samples will be stored frozen (-70C or below) and discarded at the Testing Facility - upon requestofthe Sponsor. 001109 418-015:PAGE C5 Protocol 41P8a.g0e1s5 ShippingInstructions: Samples to be analyzed will be shipped (frozen on dry ice) to: 3KrMisEJn.vHiarnosnmeenn,tPahl.TDe.chnology and Safety Services 935 Bush Avenue Building 2-36-09 St. Paul, Minnesota 55133-3331 Telephone: Telefax: (612) 778-6018 (612) 778-6176 The recipient will be notified in advance of sample shipment. DISPOSITION: Parrteicplaerweidllfboermruelattuimoends twoillthbeeSdtiusdcyaMrodneidtaotrathtetTheestpirnegviFoaucsilliytyc.itAeldlardedmraeisnsinugpobunlk test completion of all workwith the test ariicle. TESTSYSTEM: `SpecianedRsea/soSnftorrSelaecitionn: TbehceauCsrei::C1D)tBhRisVsAtrFa/iPnloufsrat(wSapsraugsueed-Dianwtlheey)rerpartowduacstisveeleacntdeddeavsetlhoepmTeesnttaSlytsotxiecmity asrttuidcilees;is2p)hahirsmtaocrioclaolgdiactalalaynadcteixvpeeirnitehnecespeexicsitesatatnhde Testing strain. Facilty"; and 3) the test Number: Initial population acclimated: Population selectedforstudy: 36 virgin female rats. 24 mated female rats (8 per dosage group). BWodyeiganhdAgte: wFheimcahletirmaetsthwielyl wbiellobredeerxepdecttoehdavtoebbeodatylweeaisgth6t0sodfa2y0s0ofgatgoe.22A5ctguealacbhodaty rweeciegiphtt,sawtill be recorded the day after receipt and will be documented in the raw data. The weight range will be included in the final report. Sex: Female be used rats will only as be given breeders tahnedteasrtearntoitclce.onsMiadleererdatspoarfttohfethseamTeesstoSuyrscteeamn.d strain will cre 001110 418-015:PAGE C6 Protocol 41P8a.g0e1s5 Source: Charles River Laboratories, Inc. "The rats will be shipped in filtered cartons by air freight and/or truck from Charles River Laboratories, Inc., to the Testing Facility. Identification: EGoeneration: Rats are permanently identified using Monel self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats are given unique permanent identification numbers upon assignmentot 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. EG1eneration: Pups will not be individually identified during lactation; all parameters will be evaluated in termsof the litter. ANIMALHUSBANDRY: All cage sizes and housing conditions are in compliance with the Guideforthe Care. and UseofLaboratory Animals`. Housing: EGo eneRarts/Fa1GenteratiionoLitners: Fo generation rats will be individually housed in stainless steel, wire-bottomed cages. except during the cohabitation and postpartum periods. During cohabitation, each pair of rats will be housed in the male rat's cage. Beginning no later than day 20 of presumed gestation, Fo generation female rats will be individually housed in nesting boxes, except during collection intervals for urine and fecal samples. During these postpartum period. collection intervals, the female rats will be housed individually in metabolism cages. Each dam and delivered litter will be housed in a common nesting box during the. 001111 418-015:PAGE C-7 Protocol 418.015 Page 7 Nesting Material: Nesting material (bed-0'cobs) will beprovided. Bedding will be changed as often as necessary to keep the animals dry and clean. Analyses for possible contamination are `conducted annually and documented in the raw data. RTooemAimr,peraatndHuumirditey: "The animal room is independently supplied with at least ten changes per hourof 100% fresh air that has been passed through 99.97% HEPA fitters (Airo Clean room). cRoonsotmanttelym.perRaotoumrehwuimllidbietymawiilnltaalisnoedbeatm6on4itFor(e1d8cCo)nsttoa7nt9lyFa(n2d6mCa)inatnadinmeodniatto3r0ed% to 70%. Light An automatically controlled 12-hour light: 12-hour dark fluorescent light cycle will be maintained. Each dark period will begin at 1900 hours EST. Rats will be given Certified Rodent Diet #5002 (PMI Nutrition Intemational) available ad libitum from individual feeders. Water Water will be availablead 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 than 1.2 ppm chlorine bacteriostat; processed at the time of analysis. wateris Water is expected analyzed tmoocnotnhtlayinfonropomsosribele bacterial contamination and twice annually for possible chemical contamination. Contaminants: Neither the Sponsor nor the to be present in the certified Study Director is diet, the drinking awareofany potential contaminants likely water or the nesting material at levels that would interfere with the resultsof this study. Therefore, no analyses other than those routinely performed by the feed supplier or those mentioned in this protocol will be conducted. 001112 418-015:PAGE C8 Protocol 4P1a8g0e15 COHABITATION cUopmopnutaerrri-vgale,nmeraalteeadnrdafnedmoamleunriattss. wiAlfltebreaacscsliimganteidont,o 3i6ndviivrigdiunalfehmoaulseinrgatosnwtilhebbeasis of dsuerliencgteadccfloirmsattiuodny.onThtheefbeamsailsoerfatpshwyislilcableaapspseiagrnaendcteoadnodsabgoedygrwoeuipgsht(s12refceomradleed rats peprrodcoesduargees agrnodupt)rebataesdedwitohn eciotmhperuttehre-vgeehniecrleatoerdth(ewetiegshtta-rotridcleerefdo)r 4r2anddaoymsizpartiioronto cohabitation. Within each cohabitation dwoitshagberegerdoeurp,macloensreactsu,tiovneeomradelre wil rat be used to per female assign female rats to rat. The cohabitation pineraiosdmewiallrcoofnstihsetvoafgianamlacxoinmteunmtosfafnidv/eodraaysc.opuFleamtaolrey praltusgwoibtshesrpveerdmiantsoiztouawiolbsbeerved considered to be at day 0of presumed gestation and assigned to individual housing, Eight each mated female dosage group. rats (those with Any remaining confirmed evidence of mating) will be female rats with or without confirmed assigned evidence to of mcoahtaibnigtatthiaotnwweirllebaesssaicgrniefdicteod eainthderdoifstcahredetdrewaittehdougtrofuuprtsheorr etvhaelucoanttiroonl agtrtohuepdpirsicorrettioon of the Study Director and the Study Monitor. Day 1 which of all lpaucptastiionna(lpiotsetrpaarretuimn)divisiddueaflilnyewdeiasghtehde day of birth and is (pup body weights also will the first day be recorded on after all pups in a litter are delivered and groomed by the dam). On day 4 postpartum, liter, where possible. litters Pups will not be culled selected to for cfiovnetminauleedpeuvpasluaantdiofniwviellfebmealsaecrpiufpiscepdevria dbeucfafpeirteadti1o0n;%tfhoermlaulnigns) (wislalvbeed cionlBloeucitend'sfrsoolmuttihoen)fiarsntdtetnhecullilveedr (psuapvsedfrionmneeuatcrhaldosage gporsosuipbl(eirfruetsupreecthiivsetoofpalitthtoerl)ogdiectaelremvianleudattioonb.e Ratemdaaiyn4inpgosctupllaerdtupmupasndwilplrbeseesravcerdiffiocred via decapitation and discarded without necropsy evaluation. ADMINISTRATION: RR outee andasfoo r Chon ice: The oral (gavage) route was selected for administration in the developmental and use because: 1) this was the route reproductive toxicology studies; and of 2) itis one of the possible routes of human exposure. 01113 418-015:PAGE C-9 Protocol 4P1a80g1e5s Meat nd Fh reqo uend cy: EoGeneration FemaleRats: Female rats cohabitation wil be given the test article once day beginning 42 days until day 0 of presumed gestation (confirmed evidence of prior to mating, such as osibtus)e.rvFateimoanloefrsaptserwmialtnooztobaeigniavesnmethaertoefstthaertivcalgeinoarltcheonvteehnitcsleorona dcaopyu0laotforpyreplsuugmeind gestation. Dosages will be adjusted daily for body weight changes and given at approximately the same time each day. E1GenerationPups: eFx1pgoesneedrattoiotnheptuepstsawritlilcnleotdubreindgirmecattley mgaivlegnetshteattieosnt (airntiucltee,robuetxpmoasyurbee) poorsvsiiablmyatemal milk during the lactation period. Ratfior DoosangeaSelelctieon: Dosages were selected on the (Argus Research Laboratories, basis Inc., of a previous study Protocol 418-008). conducted with the test article DCosaogenLevcelse, ntratanidVoolnumess: CTs [owen[ 0 [5[samosammmomm|n [oe oo 1 om |&[eomonssmmmone|n LoToTe1 om [[eusoncommmm|m Tetestarc wbconsicred 100%urs ohsupe ofdosagecacao. TM ESTSE ,ANA ALYSS ESAU ND REME -FoN GENT ERATS ION: All Periods: Atleast twice daily. Clini andlor GeneralAppearance: Acclimation Period: Atleast once. 001114 418-015:PAGE C-10 Protocol 4P1a8g.e01150 Dosage Period: Tawppircoexidamialyt.elPyrioonretohoaudrmipnoissttdroastiaogne.and once All Other Periods: Once daily. Maternal Behavior: aDbanyosrm1a,4l,b7e,h1a0v,io1r4wailndbe21repcoosrtdpeadrtduami.ly.Observed CalpipnriocparlioabtseerbvyatthieonSstumdayyDbireercteocroradnedd/omroSrteudfyreMqouneinttolry.than cited above,if deemed Body Weights: Acclimation Period: Dosage Period: Atleast once. Daily. All Other Periods: Sacrifice: Daily. Terminal weight. FeedConsumptionValues (recorded and tabulated): Acclimation Period: Atleast once. Dosage Period: All Other Periods: Weekly to cohabitation. aDanidly1d4urpoisntgpparrteusmu.meFdegeedstcaotnisounmapntdiodnaywisll1n,o4t,7be,: 10 teaxbpuelcatteedd tahfatterpduapys 1wi4llpboesgtpianrttoumc,onwshuemneitmiastemal feed. Freepeldenciosnhstuhmepfteieod.n vTahleuesseminatyerbvaelsrewiclolrndoetdbmeotraebuflraetqeude.ntly if itis necessary to MatingPerformance: Mobasteirnvgawtiilolnboefesvpaelrumaatetdozdoaialyindaursimngeatrhoefcothhaebivtaagtiinoanl pceornitoedntasndancdoonfriarmceodpublyatory plug observed in sit. 001115 418-015:PAGE C-11 Protocol 4P1a8g.e01115 NaturalDelivery: Female rats will be evaluated for: Duration of Gestation (day O of presumed gestation to the day the first pup is observed). Litter Size (defined as all pups delivered). Live Litter Size (live bon pups only). Pup Viability at Birth. `PharmacSoamkplieCnoleletctiiocn: Urine and FecalSamples: Female rats will be housed individually in metabolism cages for collection of urineand fecal samples for the following intervals: one day prior to initiation of cohabitation to the following moming and days 6 to 7, 14 to 15 and 20 to 21 of presumed gestation, as well as days 21 to 22 postpartum. Following each 24-hour collection interval, samples will be collected into centrifuge tubes, placed on dry ice and stored frozen (-70C or below) until shipment for analysis. In the event that a dam begins to deliver before completionof urine and fecal sample collection on day metabolism cage 2a0ndtopdlaacye2d1ionfapnreestsiunmgebdogxeswtiatthiosunf,fitchieendtabmedwdiilnlgb.e removed from the BloodSamples: Blood samples will be collected from eachof the matemal rats following removal from metabolism caging (prior to administration) on each of the following days: on the day cohabitation is initiated (priotor cohabitation), and on days 7, 15 and 21of presumed gestation, as well as on days 14 and 22 postpartum. The timeof blood collection will be recorded in the raw data. 001116 418-015:PAGE C-12 Protocl 4P1a8g.e01125 On all days of collection except day 22 postpartum, blood samples (approximately 1 mL each) will be collected from the orbital sinus. If necessary, `whole blood may be collected from data. On day an 22 paotsttepmaartteumsi,teb;liofsodo,stahmeplaelste(maaptpersoixteimwialtl eble4ydmoLcuemaecnht)ewdillinbtehe raw collected via the inferior vena cava. Blood will be collected and transferred into serum separator tubes. The samples will be spun in a refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (70C or below) until shipment to the Sponsor for analysis. `Shipping Instructions: shipment. All samples will `Samples will be be maintained frozen shipped on frozen on (-70C dry ice or below) until shipment for analysis. via overnight mail. A packing list will be included address. with the samples and sent Both the recipient and the to Kris Study J. Hansen, Monitor will Ph.D., at the be notified in previously cited advanceof sample METHODOFSACRIFICE -FoGENERATRIAOTSN: Rats will be sacrificed by carbon dioxide asphyxiation. NEC- FR o GEO NERAPTIOSNRAY TS: Gross lesions will be evaluation (a table of retained random in neutral buffered 10% formalin for possible units will be used to select one control group future rat from which all tissues examined at necropsy will be retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions). Unless. specifically cited below, all other tissues will be discarded. Rats Not Selected for Continued Evaluation: Any remaining female rats with or without confirmed evidence of mating that were assigned to either ofthe sacrificed and discarded treated without groups further or the control evaluation at group prior to the discretion cohabitation of the Study will be Director and the Study Monitor. RaDtesNlotiveraiLinttger: Rats that do not deliver a litter will be sacrificed on day 25of presumed gestation and examinedfor gross lesions. Uteri will be stained with 10% ammonium sulfide to confirm the absenceof implantation sites. 001117 418-015:PAGE C-13 Protocol 4P1a8g.e01153 `Scheduled Sacrifice: Oannddbalyoo2d2spaomsptlpaerctoulml,ecftoilolno,wfiengmatlheerfaitnaslwiclolllbeectsiaocnriifnitceerdv,alafnodr uarignreosasndnefcercoalpssyamofpltehes thoracic, abdominal and pelvic viscera wil be performed. The number and distribution of implantation sites will be recorded. A liver section (fight lateral lobe) from each dam will be collected, frozen and stored (70C or below) until shipment to the Sponsor for analysis. DS amu s wirthv Noivin Pupgs: Dams with no surviving pups wil be sacrificed after the last pup is found dead, missing or presumed cannibalized. Prior to sacrifice, a blood sample (approximatel4y mL) will be collected from the maternal rat via the inferior vena cava and transferred into serum separator tubes. The sample will be spun in a refrigerated centrifuge. The serum will be transferred into a polypropylene tube labeled with the study number, animal identification, date of collection, study day and collection timepoint. The sample will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. A gross necropsyofthe thoracic, abdominal and pelvic viscera will be performed. A liver section (right lateral lobe) from each dam will be collected, frozen and stored (70C or below) until shipment to the Sponsor for analysis. Postpartum data for these dams will be excluded from summary tables. Rats Found Dead or Moribund: Rats that die or are sacrificed becauseof moribund condition, abortion or premature odebisievrevraytwiiolnl bisemeaxdaem.inTehdeforrattshweilclabueseexofamdienaetdh oforrmgorroisbsulnedsicoonsn.ditAiolnivoenr stehcetidoany(trhieght lateral lobe) from each dam will be collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Pregnancy status and uterine contentsoffemale rats will be recorded. Aborted fetuses andor delivered pups will be examined to the extent possible. Uteri of apparently nonpregnant rats will be stained with 10% `ammonium sulfide to confirm the absenceof implantation sites. p-- . All samples will be maintained frozen (-70C or below) until shipment for analysis. `Samples will be shipped frozen on dry ice via ovemight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. 001118 418-015:PAGE C-14 Protocal 4P1a8g.e01154 STS, A si NTS - F1 Viability: Postpartum Period: Litters will be observed for dead pups at least twice ddaaiillyy.. The pups in each litter wil be counted once ClOinicbal serv andlaor Gtenei ralAoppen arans ce: Postpartum Period: Once daily. Clinical observations may be recorded more frequently than cited above, if deemed `appropriate by the Study Director and/or the Study Monitor. BodyWeights: Postpartum Period: Days 1 (birth), 4, 7, 14 and 21 postpartum. Sacrifice: Terminal weight. F SACRIFICE - F1 ION RATS: As previously cited for Fo generation rats. NEC- FR 1 GEO NERAPTIOS NRAY TS: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation (a tableof random units will be used to select one control group rat of each sex from which all tissues examined at necropsy will be retained, inorder to provide control tissues for any possible histopathological evaluationsof gross lesions). Unless specifically cited below, all othertissues will be discarded. Caps and labeled tubes will be weighed (combined weight, to the nearest .001 gram) before and after retentionofpooled pup samples. These weights will be documented in the raw data, and copies of these weights will be included with the packing lst prior to shipment 001119 418-015:PAGE C-15 Protocol 4P1a8g.e01155 PupsFoundDeadonDay1Postpartum sPtuaptsustahtatbidriteh.beTfhoereleuxnagsmiwnialtiboenroefmtohveelditaernfdorimpmueprvsieabdiliintywawitlelr.bePeuvaplsuawittehd lfuonrgvsitatlhat S`ainndk twoilhabveeiddeinetdifsiheodrtalsysatfitlebrorbnir;thp.upPsupwisthwiltuhnggsrotshsaltesfilooantswwililllbebeidpernteisfeiredveadsilniBvoeubionm',s Spoolsustiibolneffourtpuroesseivballeuaftuitounr.e evaluation. All lungs will be preserved in Bouin's solution for PupsFoundDeadorMoribundonDays2to21 Postpartum: lPeuspisonfsoaunnddfodreatdheorcasaucsreifoifceddeabtehcaoursteheofmomorriibbuunnddictoyndwiitlilobne. examined Pups with for gross gross lesions fevoaulnudatoinond;agyrsos2stole4sipoonsstopfaprtuupmswfiollunbde opnredsaeyrsve5dtion 2B1oupions'tspsaorltutuimonwilflorbpeopssriebsleervfeutdurine neutral buffered 10% formalin. FBoourianl'lspsuoplsutfioounnfdordpeoasdsiobnledfauytsur2eteova4lupaotsitopna.rtFuomr,aallllpluupnsgsfwoiullndbedepardesoenrvdeadysin5 to 21 postpartum, al lungs will be preserved in neutral buffered 10% formalin for possible future evaluation. Pups Not r Conti ion -Day 4 P Allilveprsupwisllcbulelecdololnecdtead4yfropmostthpeafritrsutmtewinllpbuepssaccurlilfeidce(divmieaspdeecctaipvietoaftiloitnt.er)Alflrloumngesacahnd dosage group; the will be individually rleutnagisnewdillinbeneuitnrdiavlidbuuaflfleyrerdet1ai0n%edfionrmBaoluiinn'fsorspoolsutsiiobnl,e and the future livers evaluation. Remaining culled pups will be sacrificed and discarded without evaluation. `Scheduled Sacrifice: On day 21 postpartum, blood samples wil be callected from all remaining pups on study. lBilttoeor)dasnadmtprlaenssfweirlrebdeicnotlolseectreudmvsiaeptahreatinofrertiuobrevs.enTahceavsaamfprloemsewaicllh pup, pooled be spun in a (per refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (70C or below) until shipment to the Sponsor for analysis. The pups will be examined for gross lesions. The liver from each pup will be collected, pooled (per liter), frozen and stored (-70C or below) unti shipment to the Sponsor for analysis. 001120 418-015:PAGE C-16 Protocal 4Pa1g8e01156 `AlSlasmapmlpelseswilwliblle bsehimpapiendtafirnoezdenfroonzednry(i7c0e vCiaoorvbeemliogwh)tumntaiill.shAippmaecnktinfgorlaisntawliylslisbe.. included address. wBitohththtehesarmecpilpeisenatnadndstehnettoStKurdiys MJ.onHiatnosrewni,ll Pbhe.Dn.o,tiaftietdhein pardevviaonucselyocfisteadmple shipment STATISTICAL EVALUATION: Aapvperropargieastea.ndAdpdeirtcieonnatlagpersocweildlubreescaalncdu/loarteadn.alLyistteesr vmaalyuebsewiplelrbfeorumseedd,iwfhdeereemed appropriate. DATA ACQUISITION.VERIFICATIONANDSTORAGE: DDiarteactwoirllabned/hoarnadp-paronpdr/ioartceommpauntaegre-rmeecnotrdpeedr.soRnneeclorwditshwiinll2b1edraeyvsiaefwteedr gbeynetrhaetiSotnu.dyAll obreigbionaulnrdecaonrddsinwdiellxebde. sAtorceodpiynotfheallarrcahwivdeastoaftwihllebTeesstuipnpgliFeadctioltyh.eASllpoornisgoinralupdaotna will request. Preserved year after mailingof tissues will be stored the draft final report, aafttetrhewhTiecshtitnigmeFatchieltSypaotnnsoorchwialrl gbee for one contacted to determine the disposition of these materials. 001121 418-015:PAGE C-17 Protocol 4P1a8g-e01157 REC TOO BE MR AIND TAINS ED: Protocol and Amendments. Test Article, Vehicle and/or Reagent Receipt, Preparation and Use. ARnainmdaolmiAzcaqutiisointiSocn.hedules. TMraetaintgmeHnitst(ofryp.rescribed byStaffVeterinarian). GCleinneicraallOCbosmermveanttiso.ns andor General Appearance. Tissue and Sample Collection, Processing and Shipment. Cap and Labeled Tube Weights. Body Weights. Feed Consumption Values. Natural Delivery Observations. Litter Observations. GOrrogsasn NWeecirgohptssy(iOfbrseeqruviraetdi)o.ns. PSthuodtyogMraaipnhtsen(ianrceqeui(rreodo).m and environmental records). Feed, Water and Bedding Analyses. Packing and/or Shipment Lists. KPEEYRSONNEL: Executive Director of DireofcResteaorcrh: Research: Mildred S. Christian, Ph.D., AlaMn. Hoberman, Ph.D., DABT Fellow, ATS Associate Director of Research and Study Director: Raymond G. York, Ph.D., DABT Director Director of of Laboratory Operations: John F. Study Management: Valerie A. Bamett, Sharper, B.S. M.S. Manager of Animal Operations and Chairperson, Institutional Animal Care and DiUrescetoCroomfmOiptetreaet:ioDnesnaandC.CoLmepblo,iaVn.cMe.:D.Barbara J. Patterson, BA. Consultant, Veterinary Pathology: W. Ray Brown, D.V-M., Ph.D., ACVP ce1iz2 418-015:PAGE C-18 Protocol 4P1a8g.e01158 FINALREPORT: AbecfoimnpalriezhedenfsoillvoewidnrgafctofnisnualltraetpioorntwwiitlhbteheprSepopnasroerd.onThcoemrpelpeotritownilolfitnhcelusdteudtyheand will following: `Summary and Conclusion. Experimental Design and Method. Evaluation of Appendices: Test Results. Figures, Summary and Individual Tables Summarizing the Above DGaLtPa,CPormoptloicoalncaendStAastseomceinatt,edReApmoertnsdomfenSutpspoarntdinDgevDiaattaio(nisf,apSptruodpryiDaitree)ctaonrd's QAU Statement. i TIONAL ST : TInhsetitpurtoiocneadluArneismdaelsCcrairbeedanidn tUhsisepCroomtomciotltehea.veAbleeprnorceevdiuerweesddbesyctrhiebeTdesitnitnhgisFapcriolttoyc'osl that involve discomfort, dsitsutdryesasnoirmaplasinwitlol be the conducted animals. in a manner to avoid or minimize nTehceesSspiotnysfoorr'scosnidguncattiunrge tbheilsoswtuddoycaunmdentthse ftahcetftahcatttthhast iinsfnoortmaatniounncnoencceesrsnianrgythe duplicative study may be obtained from the Sponsor. No altemative (in vitro) procedures were available for meeting the stated purposes of the study. 001123 418-015:PAGE C-19 Protocol 4Pa1g8e01155. REFERENCES: 1. CThersitsst.iaEn,nvMi.rSo.nmaenndtaVloyPtreokt,ecPt.iEo.n (A1g9e82n)c.y,IWnaVsihviongRteporno,duD.cCt.ivNeatainodnaMluTteacghenniiccailty Information Service, U.S. Departmentof Commerce, Springfield, VA 22161. 2. Cnharlitsrteixaon,neM(.PS.ro(c1e9e8d4i)n.gsRoefpNraoldturcetixvoenetoSxiycmitpyoasnidumt,erNateowloYgoyrekvaAlcuaatdieomnsyooff Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the hares River Cri:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base providedby Argus Research Laboratories, Inc.) 4. Institute of Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski, E. (1964). Farbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Rate. Arch. Pathol. Exp. Pharmakol. 247:367. 001124 PROTOCOLAPPROVAL: FOR THE TESTING FACILITY Gre---- Alan M. Hoberman, Ph.D., DABT Director of Research G. York, PhD', DABT Associate Directorof Research `Study Director Dre Chile Dena C. Lebo, V.M.D. Chairperson, Institutional Animal Care and Use Committee FOR THE SPONSOR Was JGun Marvin T. Case, D.V.M., Ph.D. Study Monitor | 418015PAGE C20 Protocol 4P1a8g.e0105 (6- wov-l Date 16-wov98 Date 1 Nev 28. Date 17 th 28 Date 001125 418-015:PAGE C-21 ATTACHMENT 1 `SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE 001126 ATTACHMENT 1 418-015:PAGE C-22 ProtocPolag4e181001152 STUDYSCHEMATIC PHARMACOKINETIC RECOVERY STUDY" Dossuargaet CoEnnsose' SScercarduiicees FoRanE ss E PPuemraetnag 2a) | Copneprractdon| [r= Gpersetastuimoend Perod | PLoascpiaavrouny = Ea. E DFoorsaadgdeitPieorniaoldd.etails see "Tests, Analyses and Measurements" sectionofthe b. prFootgoecnoelr.ation femaleratswil receive test articleorvehicle until mating is confirmed (day 0 of presumed gestation). 001127 ATTACHMENT 1 418-015:PAGE C-23 ProtocoPlag4e182.o0f125 SCHEDULE 17NOV 98 23NoV 98 04JANSSPM-09JANGIAM 05 JAN 99 03 JAN 99 26 JAN 99 03FEB 99 29 JAN 99 06 FEB 99 30 JAN 99 03 FEB 99 15 FEB 99 - 23 FEB 99 16 FEB 99-24 FEB 99 29 JUN 99 Animals Arrive - Acclimation Begins. cDoohsaabigteatPieornioundti-l FceomnafliremReadtmsat[4i2ngda(ydsaypr0ioorf to presumed gestation). Cohabitation Period. FLiarsstt PPoossssiibbllee DDaayy 00 ooff PPrreessuummeedd GGeessttaattiioonn. First Possible Delivery (Day 21 of presumed gestation). Last Possible Delivery (Day 25 of presumed gestation). First Last Possible Possible Day 4 Day4 Postpartum Postpartum Culling, Culling. First Possible Day 25 of Presumed Gestation Female Sacrifice. Last Possible Day 25of Presumed Gestation Female Sacrifice. Scheduled Sacrifice - F1 Generation Pups (Day 21 postpartum). Scheduled Sacrifice - Fo Generation Dams. (Day 22 postpartum). Draft Final Report. a. The study initiation date is the day the Study Director signs the protocol. 001128 418-015:PAGE C-24 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 001129 418-015:PAGE C-25 MATERIAL SAFETY DATA SHEET 34 Center St. Paul, Minnesota 55144-1000 1-800-364-3577 or (612) 737-6501 (24 hours) Copyright, ALL rights r19e9s8e,rveMdi.nnesCootpayiMnignianngd/aonrddMoawnnulfoaacditnugrinofg Company. this inforaation for the purpose of properly utilizing SM products is 1) tahlelowiendforpmraotviiodnedisthacto:pied in full with no changes unless 2) npreiiotrheragrtheeemecnotpyisnoorbttaheineodrifgrionmalSMi,s and resold or otherwise distributed with the intention of earning a profit thereon. DIVISION: 3M CHEMICALS TRADE NAME: FC-95 FLUORAD Brand Fluorochemical Surfactant 1D NUMBER/U.P.C.: 98-0207-0103-7 00-51135-09054-1 98-0207-0104-5 98-0211-0888-5 2ZF-0002-1044-1 00-- 51135--09362--7 98-0211-3916-1 SISUSPUEERDS:EDEJSa:nuaNroyvem29b,er190S9,8 1997 DOCUMENT: 10-3796-9 00-51135-09055-8 00-51135-02311-2 1. INGREDIENT PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SSUULLFFOONNAATTEE............ POTASSIUM POTASSIUM PERFLUOROALKYL PERFLUOROALKYL SULFONATE...... SULFONATE...... POTASSIUM PERFLUOROALKYL SULFONATE...... C.A.S. NO. 2795-39-3 82 3~28947210--499-93 -633 60270-55-5 2 3872-25-1 1 PERCENT - 86 -8 -7 -6 -3 2. PHYSICAL DATA BOILING POINT:.......ceucunenns N/A VAPOR PRESSURE:..........eeuees NIA VAPOR DENSITY:..........cceeeees N/A EVAPORATIONRATE:.............. N/A SOLUBILITY IN WATER:........... slight SPECIFIC GRAVITY:.............. CA. 0.6 Hater=i (Bulk) PERCENTVOLATILE:..........cc0n 0% VISCOSITY:.....uevnennreencnnss NI(D0.1% Aqueous) MELTING POINT:......covvvnennn. NID APPEARANCE AND ODOR: Light colored, free flowing powder. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 001130 418-015:PAGE C-26 JJaonsu:aryFG-238,5 F1L9U8O8RAD Brand Fluorochenical Surfactant age 2 37 FIRE AND EXPLOSION HAZARD DATA FFTLLUAAASRAHUBPBLOLIEENTLL:II.NM.II.TTSSoc--oeLUsEELLei:e..e..e....n..c..e..e.. NNOIDAS NIA ATOTGNITION TEMPERATURE1S.... N/A EXTTIaNtGeUrISHCIaNrGboMnEDdIiAo:xide, Dry chemical, Foss SPHI EeCIlALvAeFIREPprrFeaoIsntGsdeHusc.TrteIiNavGroeroPuRcnpOldrCoeEtsrDhsaiUusnRr,gEe,S:Wdaeiimnascnltdudaibnnrdgasltehghesli,snegtf,aicpspeaerlmaafts-ukcs,o,ntsanbidunnekde,r cost O BrtheP tive covering for exposed aress of the head. UNA USUArL FIaRnEdoAuNsDDEeXcPoLpOoSsIiOtNioHnAZAsReDcSt:ion for products of combustion. REACTIVITY DATA STABILITY: Stable INNCoOtMPAapTpIlBiIcLaIbTlYe.-MATERIALS/CONDITIONS TO AVOID: WAZARDOUS POLYMERIZATION: Hazardous polymerization Will not occur. VAZoCAiRvDnoOrUiSeM,oDnEoTCtoOsxRdiPecOSVIaaTnpdIoOrNCsa,PrRbOoGDnaUsCeDTsiSo:xoirdeP,arOtxiicduelasteosf.Sulfur, Kydrogen 5. ENVIRONMENTAL INFORMATION Tr Tr SPICGhLnsLeenroRsEueSn.PdOpNroSreEc:waautteironstofarvoomidothdeurstisnegc.tioCnAsU.TIONV!acuAumv,acuuusme oo onition source. Clean up residue with water. wcelteanSewreecpoiunlgd Place in an APproved metal container. Seal the container. REDaCFrO7oM7nM6meEotsNosDfeErsDetlheetDahIseSletPoOwStecAsooLtu:lwdaEtCeSrrDewsauoylrst LCionSrOaseqcwuoeanrtc.iecntrDcaootnicnooentn.trusaetIinocinnisneprrgaortdeeuactteisrn otrhan an mteaetreonraitan4li.ave:oCrombcDuiossmtpmioeosrneciaoplfrodfwuaacscttiselitwpyirloldiuncitnthceliundperaesfHFea.nccieliDtoiyfspoapsecarlommitbtuesdtibtloe 0011, 31 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - approxinatdiy {ier JJaSDnSu:aryFG2-99,5 1F9L9U8ORAD Brand Fluorocheaical Surfactant 418-015:PAGE C-27 Pace 3 5. ENVIRONMENTAL INFORMATION (continued) accept chemical waste. ENVREBIaoRinEOrRiiNnE1eNAiTrAAS itL)irDcsu ATmhAgF:/i(n lsL;hepoL4enCBSi-OHs,r.saFacEtrChoS0ec,ahdirDMuaispn)hn=no8iwsa(PmMiga/sglen,pahaR=laei5sn0bopsrwg/olmT;rsoluaCtsO()DS==a3.l80m0omg/l, Gig; 80020 = Nil. RE`GVUoLlAaTtOiRleY VOC Less OHIr2Ng0FaOnR&iMcAETxICeOomNsp:tpouSnodlsv:entNs/:A. N/A. SSeifnocreeredgisuploastailo.ns vUa.rS.y, EPcoAnsHualztardaopupsliMcaasbtlee rNuemgbuelrati=onsNonoer a(uNtohtorUi.St.ies EPA Hazardous). TTShCiAs,prEoIdNEuCcSt, coCOmSpLl,iesAICWSi,thMItThIe acnhdemiKcoraela.registration requiresents of EOPCRREA HWaAzZAARRDD: CLNAoSS:PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes "6.SuGESTED FIRST AID : Tm EYETaCnOeNdTiAaCtTe:ly flush eyes minutes. Get immediate wmeidtihcallargaetteanmtoiuonnt.s of ater for at least 15 SKI`NTamCeOdNiTaAtCeTl:y flush skin Sontaninated clothing. wIifthirrliartgaetioanmoupnetrssisotfs,watcearl.l Raempohvyesician. Wash Contaminated clothing before reuse. INHTtALAiTgInOsN/:syaptons occur, signs symptoms continue, craelmlovea ppheyrssiocniafno. fresh air. If IFrSiWnAkLLOtWoED:glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION or o or - EYEAvoPiRdOTEeCyTeIOcNo:ntact. Wear vented goggles. 001132 Abrevistions WD | Wet Determined NIA Not Applicable CA. Approximately 418-015:PAGE C-28 MJSaDnSu:aryFG2-99,5 F1L9U9O8RAD Brand Fluorachesical Surfactant PAE 4 7 PRECAUTIONARY INFORMATION (continued) SKIGNolPdROTsEkCiTnIOcNo:ntact. A amata pair of Wgleoavresapwpardoeprfiraotme gthleovfeosllwohweninghanmdaltienrigslt(hsi)s are DPFoaevtraoarmoisnneegnl,dapdcr:oovteerc"abtulitloysnl. irtuebPmbrseort.eAcstiUnvseeeceosgnsaearreeyonrttsomorp(eroetvohefenrtthteshkaifnonlgllcooowvneitsna)gct:shohuledad Pbeelymeatdneyloerneeiptohleyrvionfyltihdeenaellcohwlionrgidemat(eSrairaalnse:x). RECVCoeOenMtMiEwlNiaDttEheDdapVEparNreTosIp.LrAiTaItPOerNo:vliodcealsuefxfhaiucsitentvenvteinltaitliaotni.on tUosemainintaaWienll: a2lsnsoitonasdeqbuealtoew, reucseommaepnpdreodprieaxtpeosurreespilrimaittosr.y prIfoteecxthiaouns.t ventilation REASTvPeoIsiRpdAiTraObtRroYerasPtRhOibTnaEgsCeTdoIfOoNnd:usati.rborSneelacctoncoennetroaftitohne fofolcloonwitnagminNaInOtSsH aanpdproivned aPcictormdaasnkcesuwpitphlieOdSHaAirregruelsaptiiroantso:r, fhualllf--fmaacsek dduusstt aanndd mmiisstt rreessppiirraattoorr,, full-face supplied air respirator. PRrDEoVeEaNnsToItOtNhsaetOr,FougGAhrClaCynIkDEMNEoTrtAhLseSooIkaNepGESwaThnIedOnNW:autseirn.g tWhaisshprhoadnudcst.aftWearshhaenxdploisnegd and before eating. RECKOeeMpMENcDoEnDtaiSTnOeRrAGdEr:y. Keep container closed when not in use. FIRNoEntAlNaDmsEabXiPlLeO.SION AVOIDANCE: OTHNCEooRnt`asPmsRoiEknCiaAntgU:iToInOSNmAoofRkYintghIeNWFhtOiRolMbeAaTcIcuOosNi:nagndt/hoirs smporkoeducatndclaneadresToultthein forsation TofhistheHOhSa.zardous decomposition products mentioned in section 4 of WIS HAZARD RATINGS: PHEEARLSTOHN:AL2PROFTLEACMTMIAOBNI:LITXY:(Se0e RpErAeCcTaIuVtiIoTnYs:, 0section 7.) EXPOSURE LIMITS INGREDIENT VALUE UNIT PPOOTTAASSSSIIOUNM PPEERRFFLLUUOGROOAALLIKYYLL SSUULLFFOONNAATTEE...... | 00..11 MHoW/G/MM33 PPOOTTAASSSSIIUUMM PPEERRFFLLUUDORROOAALLKKYYLL SSUULLFFOONNAATTEE...... 00..11 MHGM/G/MM33 TYPE AUTH SKIN" TTWHAA sat4 YY TTWMAA a344 YY Abbreviations: NID - Not Determined N/A - Not Applicable CA - Approximately 001133 418-015:PAGE C28 SDSDaSnu:aryFC2-99,5 F19L9U8OMAD Brand Fluorocheaisal Surfactant Paces EXPOSURE LIMITS (continued) INGREDIENT VALE WNIT TIPE AUTH SKI oTASSTIN PERFLUGRGALKYL SULFOMKTE... | 0.1 MG/m3 TWA MY I - SKIcNonnNtnOiTAsnTi)IcOoNuc:sonsteLraiibsbtrueatdnieonsuanbtdostatenyhece,esoveeiirtnahdleilrcstebeyxdpomsiwuribrtoehrnb'eyY'tohru,endecmrourtSeaKneIpoNaurstriecfrueolruatretloy, itary Sontact with the substance. Vehicles can alter skin absorption. SSOomUeR.CE *OF31EXRPOeScUoRaEenLdIeMdITExDApToAs:ure Guidelines "a. HEALTH HAZARD DATA eveR3c0onEtyaec:irritation: signs/sysptoms can include redness, swelling, Pain, and tearing. SKIHiNiolnaCsONSsTkyAimCnpTt:Tormesitcaatnioninc(laufdteerrepdrnoelsosn,gedsweolrlirnegp,estaendd ictocnhtiancgt.): W"axytonbdeedabstoinreb.ed through the skin and persist in the body for an INHWAaLyATbIeONn:araful if inhaled. MTianye.be absorbed by inhalation and persist in the body for an extended Single overexposure, above recommended guidelines, may cause: SIorrreinteastsionof(Tuhpepernosreespainrdattohrryo)a:t, aicgonusg/hsiynsgptaonndssnceaenziinngc.lude IF InSWgAeLsLtOiWoEnD:is not a Likely route of exposure to this product. tIhlilsnesmsatemraiyalr.esult from a single swallowing of a moderate quantity of May be haratul 5f swallowed. MUTMAuGtEaNgIeCnIiTcYi:ty assays indicate the product is not mutagenic. Aobreviationas WD - Net Determined WIA - Not ApplicibGAle Approxiately . 001134 418-015:PAGE C-30 JMSaDnSu:aryFG-238,5 F1L99U8ORAD Brand Fluorocheaical Surfactant Pace 3. WEALTH HAZARD DATA (continued) AETPRoODUvCeTrIaVtoEg/eDnEiVcELOiPnHtEhNeTALratTOXaItNSo:ral doses below maternally toxic Teves. OTCHaElRifHoEfrAinLiTcaHePHrAoZ1apAoRsnDiottIiNoFknOnRo6Mu5An.TItOoN:Gontain any substances regulated under A Product Toxicity Sumary Shest is available. SECTION CHANGE DATES HEADING SECTION CHANGED SINCE November 05, 1997 ISSUE Aobrevistions: NID - Not Determined N/A - Not Applicable CA - Approximately TTThoHePLCIoiErnDrf,eocrtmIaNtaCiaLoUnDoIfNiGnt,hethBiUdsTateMNaOtTiesrsLuiIeaMdlI.TSEaDfITeM0t,WyAKDAENaSYtaNIOSMhPWeLsAItRERDA(NWMTASIDRESRS)A,NTisYEXbOPFeRlEiSeSvEeDdOtRo NIPEnEReRCtFHnOAeRNrMTAANtBChIeELIO3TRMYpUSrOAoRGd'EuFcItTONFEiSsTSRAfDiFEtO.RfAorUsPeA8rRTpIaiCsrUtLirAceRuslpaoPnrUsRiPpbOulSreEposOfeRoraCOndUdeRtSseEuriatiOFanbilneg for U"iisnneirq'uaseflfyemcemttihtothdhienofustehueseanudosrearapspppllikicncaoatwtilioeondng.eofanaGdivecnonptrtrohodelu,cvta,riitestoiymse eofsofsfeaWnchttiiocarhls atrhteahtat PnaertiucsuesraarvapluursptoeseThaendSSHuPirtoadbulcetfoTro duesteerr'msinmeethwohdathoefruiste iosr faiptplfiocratiaon. 3DiMunepertroor.voritsdh,eesoreaiminosftsoeiromnapstoisosonribiialnlitteeylreacttithoranotsnieclinecftotrrhoiasniacsinTafrorasanessrtfvieiorcne,mafyoIMhimtasavkeecsursetsnouomletresd. IrnefporremsaetnitoantioonbstaiansedTofriotms caomdpalteatbeanseessmayornoatccubreacaya.curIrnenatddiatsiotnh,e information in the MSDS available directly from SH. 001135 418-015:PAGE C-31 ATTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE 00113 418-015:PAGE C-32 ATTACHMENT 3 Version: 41Pr8ot(o.0co6l0N4O11V8-096185) TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE Page 1012 Test Article: Vehicle: PFOS 0.5% Tween 80 in R.O. Deionized Water A. Purpose: oTfhedopsuargpeosseusopfetnhsisiopnrsoocfedPuFreOSis taondprtohveidveehaicmleetfhoordorfaolratdhmeinpirsetpraartaitoinonto rats on Argus Study 418-015, B. General Information: 1. Aslplecsiulsypethnesipornotcoocnotlaniunemrbserwi,lltebset laratbieclleedidaenndtifcioclaotriocno,dAerd.gusEbaacthchlabel wil number, concentration, dosage level, preparation date, expiration date and storage conditions. 2a. Suspensiownilsl be prepared: X_ Daily _ Weekly 2b. _Ve_hicleDawiilllybe preparXe_d: Weekly __ For__daysofuse _For__daysofuse 3. Suspensions will be prepared at a final dosage volume of 5 mLikg. 4. safety: X_ Gloves, lab coat, goggles or safety glasses and faceshield X_ Dust-Mist Respirator Half-Face Respirator -- Ful-Face Respirator/Positive Pressure Hood TZ Tyvek SuitiApron 5. Dosage suspensions adjusted for Free base and % Purity. Yes X_ No (Calculations based on 100%) __ FreeBase __ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol. 01137 418-015:PAGE C-33 ATTACHMENT 3 Version: 418P-r0o1t5o(c0o6lN4O18V.308151 Page ar TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE NOTE: Test article will be prepared as a serial dilution from the high dosage to the low dosage. Once the final volumes are achieved, stir bars are to be aaddmdienidsttroatthieonc.ontainers; mixing should occur during sampling and/or C. Preparationof Vehicle 1. `Add the required amountof R.O. deionizedwaterto an appropriately labeled container. Heat thewater to 50C = 5C, add the required `amountof Tween 80 and mix until uniform (See TEST ARTICLE CALCULATIONS). D. Test Article Suspension Preparation: 1. To prepare the 0.32 mg/mL, Group Ill suspension, add the required amount of test article (See TEST ARTICLE CALCULATIONS) into an appropriately sized, labeled container. QS ad to the required amount with vehicle and heat the mixture to 80C +5C for approximately 30 minutes or until the TA/S dissolves. 2 Once the test article has dissolved: spin while the suspension cools. (Be sure there is a visible vortex, this will achieve the desired emulsion. This may be prepared the day before use.) 3. To prepare the 0.02 mg/mL, Group Il suspension, remove the required amount of stock suspension (Group Ill) (See TEST ARTICLE CALCULATIONS), QS ad with the vehicle and mix. 4. To prepare the 0 mg/mL, Group | suspension, add required amount of vehicle to an appropriately sized, labeled container (See TEST ARTICLE CALCULATIONS) and mix. witenby: _ Liv lo Cut A Approved by Lo Date: _fe-vevop' Clarification: _/ No __ Yes (See attached clarification form.) nitalsiDate : (leFighiom & Bop 3/3/3, 001138 418-015:PAGE C-34 PRIMED ~ ICA ArseResSearcohDLpras.brBBieoeniebes, - Teepe Gr5 awtosn PROTOCOL 418-015 Oral (Gavage) Pharmacokinetc Recovery Study of PFOS in Rats SPONSOR'S STUDY NUMBER: T-6295.14 - `Amendment 1 - 07 January 1999 1. A(nPaalgyese4sooffthPerepproatroecdolF)o:rmulations,ConcentrationofTest ArticleFormulations day prepared [Effective date: 06 January 1999] Duplicate samples (2 mL each) will be taken from the first and last preparation rather than, the first and last preparation on the ReaforsCho angne: "The preparation occurs the day before it is used for dosage administration and, according the to protocol, day 0 of gestation is the last day of dosage padrmeignniasnttr,attihoen.saSmipnlcee cdaanyno0tobfegetsatkaetnioonf itshtehelasdtadyatyhaotftphreepraartastiaorne. confirmed l eb Alan M. Hoberman, Ph.D., DABT Director of Research mn Date foc fh 07-99 lond G. York, Associate Director Study Director Ph.p., DABT Date search and J | ltmndeed ; Dena C. Lebo, V.M.D. ate Chairperson, Institutional Animal Care and Use Committee gDoes 72. 2% 92 Marvin T. Case, D.V.M., Ph.D. Study Monitor Date 001139 APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 001140 418-015:PAGE D-1 DEVIATIONS FROM THE PROTOCOL AND STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 1. All rats were administered the test article or vehicle on premating day 43, 4 January 1989. The rats should have been placed into cohabitation on premating day 42 after dosage administration. This deviation did not adversely affect the outcome or interpretation of the study because no data were lost. 2. From 23 November 1998 to 26 November 1998 (days 110 4 of the premating period), the following Fo generation female rats received the incorrectly calculated amount of test article in vehicle, resulting in the rats receiving 25% moreofthe test article than required. Dosage Groiup I] Dosage (mag/0k.g1/day) R1a37t3N8u-mb13e7r4s9 16 137-1537061 These deviations did not adversely affect the outcome or interpretation of the study because the dosages were only for the first four days outof a total of 43 days of test article administration prior to mating. eo Horm ni All deviations arg`documented in the raw data. Raythond G. York, n.0), DABT Date Associate Director of Research and Study Director 001141 APPENDIX E TEMPERATURE AND RELATIVE HUMIDITY REPORTS 001142 418-015:PAGE E-1 ARGUS Temperature and Relative Humidity Report Location: Room 15 Protocol Number: 418-015 Range of Dates: 17-Nov-1998 14:00 to 27-Nov-1998 08:55 TTanigooRange: Tota NomofaBayre: ToTotal mNeunmoob ffHbe oouarr sF:ins: Temperature || Relative Humidity %u234.72 W"e234.712 MMMeoaexnnaini(meunmS:D): Number of Points in Range (%): NNuummbbeerr ooff PPooiinnttss LHiogwh ((%%)):: a6o&s7H.0 (09) 25%"2446 (48) 232 (98.7) 235 (100.0) 03 "0.30 o0 (0..00)) Report Generated: 26-Apr-1999 at 12:33 comments: revieweosv: AiL ch oars: 4115/5 1 Cumatve by Location (40491.97) 001143 ARGUS 418-015:PAGE E-2 Temperature and Relative Humidity Report Location: Room 04 Protocol Number: 418-015 Range of Dates: 27-Nov-1998 08:55 to 30-Nov-1998 09:15 | STpaercgieetsR:arnagte: TToottaall NNuummbbeerr ooff HDoauyrss:: `Total Number of Data Points: T8e4mFpelroaTt9urFe | Rela3t0i%vetH0u7m0i%dity 7"14.99 714.99 7 75 Mean ( SD): MMeadxiiamnu:m: Minimum: NumobfPoeinrts in Range (%): NNuummbbeerr ooff PPooiinnttss LHiogwh((%%)):: 724 (09) | 451 (x48) 772433 "684.17 6s 400 75 0 000)| .0) 75 0 (1000)0) 0 0) 0 0) Report Generated: 28-Apr-1999 at 12:35 COMMENTS: Rrevieweo ey: _{(, [oh DATE: 15/55 `Cumulative by Location (v04.01.97) 001144 ARGUS 418-015:PAGE E-3 Temperature and Relative Humidity Report Location: Room 27 Protocol Number: 418-015 Range of Dates: 30-Nov-1998 08:15 to 09-Dec-1998 14:30 TSpaercgieetsR:aantge: TToottaall NNumubemroobffHDeoauyrsrs:: Total Number of Data Points: TSemFpe0ra7tu0re | Rela3t0i%ve1H0u7m0i%dity 21100 221100 22 22 Mean ( 5D): MMeadxiiamnu:m: | Minimum: NNumubemroobffPPaeaiinnrttss HinigRhan(4g)e: (%): Numberof PointsLow('): M3 os | m3 een 77234 6681s2 700 370 2 o9o|| 2oo2 (9000 9 | oo (0 Report Generated: 26-Apr-1999 at 1237 COMMENTS: REVIEWED BY: IL I DATE: Ls/ 74 `Cumulative by Location (v04.01.97) 001145 ARGUS 418-015:PAGE E4 Temperature and Relative Humidity Report Location: Room 28-29 Protocol Number: 418-015 Range of Dates: 03-Dec-1998 14:30 to 15-Dec-1998 15:44 STpaercgieetsR:arnagte: TToottaall NNuummbbeerr ooff HDaoyusr:s: Total NuomfDabtaPeoinrts: TFemlpeora7t8urFe | Rola3t0i%vetHou7m0i%dity 15.701 1457.01 145 45 Mean (2 SD): MMeadxiiamnu:m: Minimum: 707 1.1) 49.0 (35) 776054 Sa0s4 60.1 308 NNuummbbeerr ooff PPooiinnttss HinigRhan(%g)e: (%): Number of Points Low (%): 146 (100.0) 146 (100.0) 0) 0 0) 0 (0.0) 0 .0) Report Generated: 28-Ape-1999 at 12:39 COMMENTS: neveves or,ALA 7 `CumulativebyLocation (v04.01.97) 001146 ARGUS 418-015:PAGE ES Temperature and Relative Humidity Report Location: Room 27 Protocol Number: 418-015 | Range of Dates: 15-Dec-1998 15:44 to 21-Dec-1998 13:45 STpaercgieetsR:arnagie: TToottaall NNumubemroobffHDeoauyrsrs:: Total Number of Data Points: TFemlpeora1tu0re | Rela3t0i%vetHou7m0i%dity at.7 rat7 a Vis Mean (50): MMeadxiiamnu:m: Minimums: NNuummbbeerr ooff PPooiinnttss iHnigRhan(%g)e: (X): Number of Points Low (%): 708 wom| se 26) 770255 Faos 0 i uose 0`o0o || w0s o`ooon ooo | 0 00 Report Generated: 26-Apr-1999 at 12:41 COMMENTS: REVIEWED BY:LLL oats: 7h `Cumulative by Location (v04.01.97) 001147 ARGUS 418-015:PAGE E6 Temperature and Relative Humidity Report Location: Room 28-29 Protocol Number: 418-015 | Range of Dates: 21-Dec-1998 13:45 to 20-Feb-1999 15:00 TSapregceitesR:arnagte: TToottaall NNumubemroboffDeHaoyursr:s: Total NumberofData Points: Mean (50): MMeadxiiamnu:m: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): Report Generated: 26-Apr-1999 at 12:46 TSemFpleroaTtuFre Rela3t0i%vetHou7m0i%dity 146842.99 146642.99 146s 1465 00 @1 | 490 761990 satrs es 100 1466 (100.0) 1407 0 (0.0) 49 0 .0) 10 (108 (96.0) 33 on COMMENTS: revieweo ev: JffA onte: nts Cumulative by Location (v04.01.97) 001148 ARGUS 418-015:PAGE E-7 Temperature Deviations Report Location: Room 15 Protocol Number: 418-015 Range of Dates: 17-Nov-1998 14:00 to 27-Nov-1998 08:55 `Temperature Target Range: Species: rat 25NDoaw-t1e988 T0i30m0e Te6m3p8.L 2255NNoovv--11999888 00670000 6633.7411 64F to 79F Date Time Temp. H=Valus ooufratTnegemp-.Hi=gThem_peLra=tuVraelu*eFout of range - Low Report Generated: 28-Apr-1999 at 12:34 These deviations dit nt adversey ate the autome or interpretation ofthe suc: "The following deviation(s) impacted on the outcomeofthe study as described: StudyDirector: i 5 = Dategp@P90 Deviations by Location (v04.01.87) 001149 418-015:PAGE E-8 ARGUS Relative Humidity Deviations Report Location: Room 28-29 Protocol Number: 418-015 Range of Dates: 21-Dec-1998 13:45 to 20-Feb-1999 15:00 Humidity Target Range: Species: rat 30% to 70% sbSepDeoeaetctmieeieossm Ti111833m000e000 RTTmHa2o.bHn OOOffmiaimmiicoosmm DBsao0pd mmMoaaallb amSCmuiniisoem oGGroa IaaLbL GGGelelanemrmiieoosmw Gooesedo 11lga7okkb (IISenalnnttioeenm T1d5ie0oa0 TTTeeoennn nWiainieom oFaao TTeeanm SBmDhmaiiitemooae Tioo18ssm0eee0 RTTTHooA.omHn DLrliamimneiem 0oh0o 1T0o4umn BEBliiinnniie a0T%k r7miaenmn arLamurenetir me men Iffliiimnieeie BGGeieeo msToaaonnh ffliinnit oosees 7m3aanm Wx Value out of ange -High L = Value outof range - Low R.H. = Relative Humidity (%) Report Ganaraind: 28-Apr-1990 al 1253 L These dovinions didnot acversly afct he avtome a nterprtation of he cy. "The following deviation(s) impacted on the outcomeofthe study as described: Study -- ; 5 } Date: 30 AH Deviations by Location (4401.57) 001150 ARGUS 418-015:PAGE E-9 Relative Humidity Deviations Report Location: Room 28-29 Protocol Number: 418-015 RangeofDates: 21-Dec-1998 13:45 to 20-Feb-1999 15:00 Humidity Target Range: Species: rat 30% to 70% ZvaDaanntiteooww 2aniow Ti007mm0e0 1000 R7T7H05o.381MMH ZZBaainnnidwiee z1i5t0oo0 7TTasssehn SFFinnn ios 111548000000 rsseoissan niinniiie im11m00e0 TTToaeeTeHMn ZTTuuuannnieeossw 111561000000 TT1eE0sTahHm Tnuinees 21080000 rToassnh G2GaaFDFFeaeepntniieisssewsse T221i01900m0000e R017H7T7.HHH FGGiaeFFreeibriieeonnw 02o33r00o00 7715174THmH GGoirFeebbiieeens 0120000 77252emm H = Value RH. = oou frat nge -ReHliagthive HLu=mVialdue09o)ut of range - Low Report Generated: 26-Apr-1999 at 1254 /__ These deviations did not adversely affect the outcome or interpretation of the study. "The following deviation(s) impacted on the outcome of the studyas described: Study one /~ [ x . = Date: 35.905) Deviations by Location (v04.01.97) 001151 APPENDIX F STATEMENT OF THE STUDY DIRECTOR 001152 %PRIMEDICA 418-015:PAGE F-1 SS PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.14 STATEMENT OF THE STUDY DIRECTOR This final report accurately reflects the raw data obtained during the performance of the study. No deviations from the U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations; Final Rule", the Japanese Ministry of Health and Welfare (MHW) Good Laboratory Practice Standard for Safety Studies onDrugs and the European Economic Community (EEC) Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles ofgoodlaboratorypractice occurred that affected the quality or integrity of the study. Associaitned GD.irYoerckt/oPhr.[).,of-RDbsAearBchT Date and Study Director a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1988). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. c. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Communityof an (OECD decision/recommendation on compliance with principles of good laboratorypractice. Official Journal of the European Communities: Legislation. 32(No. L 315; 28 October): 1-17. 001153 APPENDIX G QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT 001154 r2 PRIMEDICA 418-015:PAGE G-1 ArguS0s8RSesheeaerhcyh DLiavbeo.ratBouriielsd.iInAncg TelephoHnoer:sh(i2e13n),4P4A3.189701404 Telex: (215) 443-8587 QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT Study Director: Raymond G. York, Ph.D., DABT Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Protocol 418-015: Oral (Gavage) PFOS in Rats Pharmacokinetic Recovery Study of Sponsor's Study Number: T-6295.14 and The draft protocol Drug Administration (foFrDtAh)isGsotouddyLwaabosraatudoirtyePdrafocrtiacdeheRergeunlcaetitoonsU,.S.JaFpaonoedse Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic. Community of an OECD decision/recommendation on compliance with principles of good laboratory practice on 18 OCT 98. Critical phases of this study were inspected 10 times; study information and raw data were audited twice (see tables 1 and 2 for dates and phases/data). The draft final report and the raw datafor thisstudywere compared and taoudUi.tSe.d FfooroadcacunrdacDyr,ugfoArdamdihniesrternacteiotno (pFrDoAto)coGloroedquLiarbeomreanttosr,yaPnrdacftoirceadherence RPreagcutliacteioSntsa,ndJaarpdanfeorseSaMfientiystSrtyuodfieHseaolnthDraungds,WealnfdarEeur(oMpHeWa)n;EGcooondomLiacboratory Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on a`cnodmp1l5iaJnUceNw9i9t,hapnridncfioprlreesvoifsigoonsodrelqaubeosratteodrybyprtahcetiScpeobnestowreoenn 2100 JMUALY999,8 22 JUL 99, and for finalization on 23 JUL 89. 001155 418-015:PAGE G-2 This study was conducted according to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on on 28 July Drugs. And 1989 on the European Economic Community (1989) acceptance by the European Economic council decision Community of an OlaEboCrDatdoreycipsraicotni/cre.ecommendation on compliance with principles of good or ( . NancyJ. Gongliewski Date Quality Assurance Manager / . 25077 free A. Zaborowski, B.S. Date Senior Quality Assurance Associate and Principal Auditor 001156 TABLE 1 CRITICAL PHASES INSPECTED 418-015:PAGE G-3 Date of inspection: 23 NOV 98 Date results reported to the Study Director and Management: 25NOV 98 Test Article Preparation Date of inspection: 01 DEC 98 Date results reported to the Study Director and Management: 02 DEC 98 Urine/FecalCollection Date of inspection: 04 JAN 99 Date results reported to the Study Director and Management: 04 JAN 99 Blood Collection Dates of inspection: 04 JAN 99, 17 FEB 99 04 Dates JAN 99, resus 17 FEB reported 99 to the Study Director and Management: Cohabitation Date of inspection: 05 JAN 99 Date results reported to the Study Director and Management: 07 JAN 99 Natural DeliverylLitter Watch Date of inspection: 27 JAN 99 Date results 02 FEB 99 reported to the Study Director and Management 001157 418-015:PAGE G4 Cul Pul pSci rifince;g Da, y4 Date of inspection: 01 FEB 99 Date results reported to the Study Director and Management: 03 FEB 99 Necropsy, Fo Dams, F; Pups Dates of inspection: 17 FEB 99, 17 FEB 99 Dates results reported to the Study Director and Management: 22 FEB 99, 22 FEB 99 001158 418-015:PAGE G-5 TABLE 2 RAW DATA AUDIT(S) `The from 07 following study information APR 99 to 13 APR 99: and raw data were audited Protocol. LPirsottoofcoplerasmoennndemleanntd.computer operator codes. AEnrriomralcordeecesipatn,dracnoddoemsizfaotricolnin,icpahlyssiigcanloebsxearmviantaitoinso.n and acclimation. IFne-eldfectornasnusamcpttiioonn.record. CNaothuarbailtadteiloinv.ery observations. LPiuttperboobdsyerwveaitgihotnss.and status. PNeucproRpasnyd.omization. Organ weights. TMiaslseuebrpeaecdkeirncgolliostnsy records. General comments. TSetmupdyermaatiunrteenaanndcreelraetciovredhsu.midity reports. Feed, water and Edit requests. bedding analyses. DDeavtiaatrieovnise.w page. KBleoyofdorcotlelsetcitnigonfadcialtiatyacnodmppuatcekrinbgalcisktus.p record abbreviations. Urine/Fecal collection data and packing lists. on 1T5hAePrResu9l9t.softhis audit were reported to the Study Director and Management 001159 418-015:PAGE G-6 The following study information and raw data were audited on 20 APR 99: Vehicle receipt, preparation and use. Test article receipt, preparation and use. Test article packing lists. Deviations. The results of this audit were reported to the Study Director and Management on 23 APR 99. 001160