Document XzeQ9jdz4jVEaRjVvB5RJpYrG

Fa 73 5)) Lary J `Summary PFOS Rabbit Teratology Sunt Numbers, 3M T-6295.10, Argus 18012 CCommpLoa&uLn,xd EPFOOS (Proflnsoirodncsatncoifwiotnhtf)r-aLroetpo2r1t7.,N98.o4-%aptmrse(loStMsDs sAend mytiwo-Ryeeuaesrtt. 2 73 oo 70S cncinogeicty sud (T-6295, Covance 6329-18) `Studs Tule: Oral (Stomach Tube) Developmental Toxicity StudyofPFOS in Rabbits ~z A3 ReonDai: 11 January 1999 = StudyMonitor Suommmfabriyn:s0(el8vseitmsehnidwcboeyhrueMhlaeavdrxlvabsmieeinonnf.ee0Cx.fps0oo.r)s1se,odP1f1r0se.2geu3n,raeanotn(dridacbrbria75)foermmhgaeaallennkso,pggdse2en2ysspisoennwgegdarrydosciuoswtlcehroareonduigahe2d0ay E e eae andehen aeisgfshentctufasawloehyreoecs2ec.xu5aarameninddnet3.ha7f5oermh1ke.e0gl.2t7.a55dl.osmacehnp.adng3gN7eoos5smceo.ympAosudingondsiefriegclraioneutdpsed.deusTcihiseoodneifcifnuefrroeeodndcae in ndyodesspeeloeve e.veee im on nee 2.5dmksaMdveoisa,rnanpdbpnyweenwigeh3ls7t5wefmreeckdtehcdaeotacsso.ecdcTuihnretehdseba2ot.r5hieoanntsdwao3.Ht7ih5gehmheiskgthgdodsogseroeluevpvsee.rles a3 probaablyismeceaonidonafdryoafd0metltahoyxiedcdaitmoys.stoiTxfihicceiattiyin.ocnresstssedwnruembfeorunodfiabotrhtio2n.s5 aanndd r3e.d7u5cmedepkupfweetiusgehstswhwiecrhe is g= 2 o3z0 steomepouind-nreltofedmastoetrntal stooxiscistkuyelaeethaelsmealdfoorsmeatvieoln.s were foundat any dose level. Thus, the on o> =i 52 cThoempfousndownaesnNtoOtEfeLouwnradstvoa1a0bmest0e.1olmggs.eintnhe irabcbi. Teruogensc NOEL in herbi wis > 375 mugtwice during the say and these samples were sen lo 2~ LF 22 TNa E eep eoy t lsousts2aepabl(oedaaynwtiearmrhyaerlionl(Bgslridoioud2op.)xLacinekdepwhtiesleoF,woslalaomdwpilhneigsghswademropelsecszolowleeucrt)ee.dcafrlTohcmedse:exaseneifrmeuammlaslaensd: Ee roanpa mimsenasHolusoAensmbaanedcnpaolancbee.notrasr(oapoeliendBegset2.eanAlifomymeiorefettssh.iss swTuehmrmeeanrosyte,cmMlpauerdscehwder9ien9ft.heoe ol report not i his sammy. . vw mTReY 6 AW 12snioe Edie 3 000650 & Epa-OTS 000811786V FINAL REPORT PROTOCOL 418-012 ORAL (STOMACH TUBE) DEVELOPMENTAL TOXICITY STUDY OF PFOS IN RABBITS SPONSOR'S STUDY NUMBER: 6295.10 FINAL REPORT DATE: 11 JANUARY 1999 000651 PROTOCOL 418.012 ORAL (STOMACH TUBE) DEVELOPMENTAL TOXICITY STUDY OF PFOS IN RABBITS SPONSOR'S STUDY NUMBER: 6295.10 TAOFBCONLTENE TS SUBJECT PAGE I. SUMMARY AND CONCLUSION 1 A Methods 1 B. Results 2 C. Conclusion 3 Il. DESCRIPTION OF TEST PROCEDURES 11 A. Conduct of Study 1 AA. Sponsor 11 A2. Testing Facility 1 A3. Study Number 11 A4. Sponsor's Study Number I AS. Purposeofthe Study [=] AS. Study Design [=] ! 000852 SUBJECT PAGE AT. Regulatory Compliance 11 A8. Ownershipof the Study 1-2 A. Study Monitor 12 A10. Alternate Study Monitor 1-2 A11 Study Director 1-2 A12. Technical Performance 2 A13. Report Preparation 1-2 A14. Report Review 12 A.15. Date Protocol Signed 1-2 A.16. Dates of Technical Performance 3 A17. Records Maintained 1-3 B. Test Article Information 3 B.1. Description 3 B2. Lot Number 1-3 B3. Date Received and Storage Conditions 3 B.4. Special Handling Instructions 3 BS. Analysis of Activity 3 C. Vehicle Information 4 CAA. Description 14 C2. Lot Number 4 C.3. Date Received and Storage Conditions 4 C4. Special Handing Instructions. 14 i 000653 SUBJECT PAGE C5. Analysis of Purity 4 D. Test Article Preparation 4 D.1. Sample Information 5 D2. Analytical Results 15 E. TestSystem 15 EA. Species 5 E2. Strain 5 E3. Supplier (Source) Is E4. Sex 5 ES5. Rationale for Test System 6 ES. Test System Data 16 E7. Method of Randomization 16 ES. System of Identification [8 F. Husbandry 7 FA. Research Facility Registration nr F.2. Study Rooms 7 F.3. Housing 7 F4. Lighting 7 F5. Sanitization 7 F6. Feed 7 F.7. Feed Analysis 8 F.8. Water 8 u 000854 SUBJECT PAGE F.9. Water Analysis 8 G. Methods 9 G.1. Dosage Administration I G2. Rationale for Dosage Selection 9 G3. Route of Administration 9 G.4. Rationale for Route of Administration 19 G5. Frequency of Administration 9 G6. Length of Study 9 G7. Method of Study Performance I-10 G8. Gross Necropsy 110 G9 Statistical Analyses 12 I. RESULTS 1-1 A. Mortality, Abortions, Clinical and Necropsy Observations 1-1 AA. Abortions 0-1 A2. Clinical Observations "3 A3. Necropsy Observations "3 B. Matemal Body Weights and Body Weight Changes 1-3 C. Matemal Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values 4 D. Caesarean-Sectioning and Litter Observations 4 E. Fetal Alterations ns EA. Summary of Fetal Alterations 1-5 E2. Fetal Gross External Alterations ne w 000655 SUBJECT PAGE E3. Fetal Soft Tissue Alterations ne E4. FetalSkeletalAlterations nz F. Satelite Rabbits I-10 REFERENCES 1-12 APPENDIX A- REPORT FIGURE Figure1. Maternal Body Weights A APPENDIXB - REPORT TABLES Table 1. Clinical Observations and Necropsy Observations -- Summary B-1 Table 2. Uterine Contents and Litter Data for Individual Rabbits That Aborted B2 Table 3. Maternal Body Weights - Summary B4 Table 4. Maternal Body Weight Changes - Summary B-6 Table 5. Matemal Absolute Feed Consumption Values (g/day) - Summary 87 Table 6. Maternal Relative Feed Consumption Values (g/kg/day) - Summary B8 Table 7. Caesarean-Sectioning Observations - Summary B-9 Table 8. Litter Observations Summary (Caesarean-Delivered Fetuses) -- B-10 Table 9. Fetal Alterations - Summary B11 Table 10. Fetal Gross External Alterations - Summary B12 Table 11. Fetal Soft Tissue Alterations - Summary B-13 Table 12. Fetal Skeletal Alterations - Summary B14 v 000656 SUBJECT Table 13. Fetal Ossification Sites - Caesarean-Delivered Live Fetuses (Day 29 of Gestation) - Summary Table 14. Clinical Observations - Individual Data Table 15. Necropsy Observations - Individual Data Table 16. Matemal Body Weights - Individual Data Table 17. Maternal Feed Consumption Values - Individual Data Table 18. Caesarean-Sectioning Observations - Individual Data Table 19. Litter Observations (Caesarean-Delivered Fetuses) -- Individual Data Table 20. Fetal Sex, Vital Status and Body Weight - Individual Data Table 21. Fetal Alterations - Individual Data APPENDX C- PROTOCOL AND AMENDMENT APPENDIXD - PILOT REPORT APPENDIX E - HISTORICAL CONTROL DATA APPENDIX F - STATEMENT OF THE STUDY DIRECTOR APPENDIX G - QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT PAGE 8-19 B-20 8-26 8-33 B48 B63 B69 B75 B-87 C-110C-32 D-1t0 D-94 E-1toE-14 Fl G-1t0G4 Pp 000657 418-012.PAGE I-1 TITLE: ORAL (STOMACH TUBE) DEVELOPMENTAL TOXICITY STUDY OF PFOS IN RABBITS ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-012 SPONSOR'S STUDY NUMBER: 6295.10 I. SUMMARY AND CONCLUSION A. Methods One hundred ten New Zealand White [Hra(NZW)SPF] rabbits were assigned to eachoffive dosage groups (Groups | through V) for the main portion of the study. Nineteen additional female rabbits were assigned to one of five dosage groups for the satellite study (three, five, three, three and five rabbits assigned to Groups | through V., respectively). The test article, PFOS, or vehicle, 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water), was administered orally (via stomach tube) once daily to female rabbits on days 7 through 20 of presumed gestation (DGs 7 through 20). Dosages of 0 (Vehicle), 0.1, dosage volume of 5 mL/kg, 1.0, 2.5 adjusted and 3.75 mg/kg/day daily on the basis of were administered individual body at a weights. `The female rabbits were observed for viability at least twice each day of the study. The rabbits were also examined for clinical observations of effects of the test article, abortions, premature deliveries and deaths before and approximately 6du0rimnignuttheespaofstterdoesaacghedpaeirliyodi.ntuBbaotdiyonweduirgihntgs twheerdeorseacgoerdpeedrioond,DaGnsd 0onacnedd6aily through 29. Feed consumption values were recorded daily throughout the study. All surviving rabbits were sacrificed by intravenous administration of Beuthanasia-D Special euthanasia solution on DG 29 and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number of corpora lutea in each ovary was recorded. The uterus of each rabbit was a. Detailed descriptions of all procedures used in the conduct of this study are provided in the appropriate sections of this report and in APPENDIX C (PROTOCOL AND AMENDMENT). 000858 418-012.PAGE I-2 `examined for pregnancy, number and distribution of implantations, live and dead fetuses and early and late resorption. The gross external alterations and sex. Visceral fetuses were weighed, examined alterations and cavitated organs for were evaluated by dissection. The brains of approximately one half of the fetuses in each litter were free-hand cross-sectioned and examined in situ. All fetuses were alterations. eviscerated, stained with alizarin red S and evaluated for skeletal Rabbits in the satellite study were sacrificed on DG 21. Blood samples were collected and centrifuged. The liver was excised, weighed and sectioned Fasestiugsneesdwteorteheexmaamiinnestdudgyr.ossFleytutosetsheaenxdtepnltacpeonstsaieblweearse dpeosoclreidbepderfolritrtearb.biAtfster `completion of were shipped sample collection, to the Sponsor for serum, liver analysis. section, fetal and placental samples B. Results Aindmtiennisatbroarttiioonnsof(otnhee 2in.5thaen2d.53.m7g5/mkgg//kdga/yddaoysdaogseaggreosuopfatnhde tneisnteairntitchlee resulted o3f.7t5hemgdo/skign/gdapyerdioosda(goen gDroGusp)2.2,T2h4e,s2e5aobror2t8i)o.nsAlococtuhrerreddoaefstersutrhveivceodmptloetion scheduled sacrifice. Increased numbers of does in the 3.75 mg/kg/day dosage group had observations rabbits in the of scant 1.0 and or no feces. Scant feces also 2.5 mg/kg/day dosage groups, occurred in one and respectively. The. three 1.0 mg/kg/day dosage group rabbit also had soft doe in the 3.75 mg/kg/day dosage group had red or liquid feces on substance in the one day. cage pan One `associated with impending abortion. No other clinical or necropsy observations related to the test article occurred. oDcocsuargree-ddeinpetnhede1n.t0,,2s.i5gnainfidca3n.t7b5omdgy/wkegi/gdhaty rdeodsuactgieongsroourpbsofdoyr wtheeigehnttilroesdsoessage pweeriigohdt (gcaailncsuloactceudrarsedDiGnsth7et2o.521a)n.dD3o.s7a5gmeg-/dkegp/ednadyendtosraegdeucgtiroonuspsifnorbotdhye entire gestation period (DGs 0 to 29) and for the gestation period after the initiation of dosing (DGs 7 to 29). Average body weights were significantly reduced on (g/day) and DGs 17 relative through 24 in the 3.75 mg/kg/day dosage (g/kg/day) feed consumption values were group. Absolute reduced in the 2.5 and 3.75 mg/kg/day dosage groups for the entire dosage period (DGs 7 to 21), and the entire period after the initiation of dosage (DGs 7 to 29) Fetal body weights (total, male and female) were significantly reduced in the 2.5 and 3.75 mg/kg/day dosage groups. Delays in fetal ossification associated with 000659 418-012:PAGE I-3 the significantly reduced fetal body weights in the 2.5 and 3.75 mg/kg/day dosage groups were evident as significant reductions in the litter averages for ossified sternal centers in the 2.5 and 3.75 mg/kg/day dosage groups and significant reductions in the litter averages for ossified hyoid and metacarpals. and significant reductions in the litter and fetal incidencesof incompletely ossified pubes in the 3.75 mg/kg/day dosage group. Cc. Conclusion Based on these data, the maternal no-observable -effect-level (NOEL) for PFOS is 0.1 mg/kg/day. The 1.0 mg/kg/day and higher dosages reduced maternal body weight gain. Dosagesof 2.5 and 3.75 mg/kg/day caused abortions and reduced feed consumption. The developmental NOEL is 1.0 mg/kg/day. Dosages of 2.5 and 3.75 mg/kg/day caused reductions in fetal body weights and delays in ossification. Based on these data, PFOS should not be identified as a selective developmental toxicant; the compound was not found to be teratogenic in the rabbit. Mited d Lot s simmnr Mildred S. Christian, Ph.D_, Fellow, ATS Date Executive Director of Research Alan M. Hoberman, Ph.D., DABT Director of Research Date A OS ond G. York, Ff PABT Associate Director orResearch and Study Director Iw:9 Date 000660 418-012:PAGE II-1 Il. DESCRIPTIONOF TEST PROC! A. Conduct of Study: AA. Sponsor: 3M Corporate Toxicology, 3M Center Building 220-2E-02 St. Paul, Minnesota 55144-1000 A2. Testing Facility: Argus Research Laboratories, Pennsylvania 19044-1297 Inc., 905 Sheehy Drive, Building A, Horsham, A3. Study Number: 418.012 Ad tudy Number: 6295.10 AS. Purpose of the Study: TNheewpZueraploasnedoWfhtihtise s[tHurday:w(NaZsWd)eSsPiFg]nepdretsoudmeetedctpraedgvnearnstefeefmfaelctesroafbbPiFtsOaSndon idmepvlealnotpatmieonnttoofctlhoesuermeborfytoheanhdarfdetpuaslacteo.nseThqiusensttutdoyewxapsosduerseigofnetdhetodoeevaflruoamte ICH in a Harmonised Tripartite nonrodent species. Guideline stages C and D of the reproductive process AS. Study Design: HTahremroenqiusierdemTernitpasrtoiftethGeuiIdnteelrinnaeti"ownaelreCounsfeedreanscethoenbaHsairsmofonrissatutdiyonde(sIiCgHn). AT. Regulatory Compliance: The study was conducted in (GLP) regulations of the U.S. compliance with Food and Drug the Good Laboratory Practice Administration (FDA)? the Japanese Ministry of Community (EEC)*. Health There and Welfare (MHW) and the were no significant deviations European Economic from the GLP regulations that affected the quality or integrity of the study. Quality Assurance 000661 418-012.PAGE 11-2 dUnoictufmiennditnegds daenrdivheadvferboemetnheprionvsipdeecdtitoonsthdeurSitnugdtyhDeicreocntdourcatonfdtthhiesTsetsutdiyngare Facility management. A8. OwneorftshehStiudpy: The Sponsor owns the study. tissues are the property of the All raw data, Sponsor. analyses, reports and preserved AS. Study Monitor: Manin T. Case, D.V.M., Ph.D. A10. Alternate Study Monitor: Andrew M. Seacat, Ph.D. A11. Study Director: Raymond G. York, Ph.D., DABT (Associate Director of Research) A12. Technical Performance: JKroihsnteFn. lBaanmdeotlta,ShB.eSr.er(,DiBr.eSc.to(rRoefseLaarbcohraAtsosroyciOapteer/aFteitoanls)Evaluation) Matthew J. Vaneman, B.S. (Laboratory Technician) A.13. ReportPreparation: Raymond G. York, Ph.D., DABT Michelle R. Rzaca, B.S. (Study Coordinator) Susan K. Bradshaw, B.S. (Data Management Specialist) Karen G. Parker, AA. (Report Administrator) A.14. ReportReview: Alan M. Mildred Hoberman, Ph.D., S. Christian, Ph.D., DABT (Director of Research) Fellow, ATS (Executive Director of Research) A.15. Date Protocol Signed: 11 August 1998 000662 418-012:PAGE II-3 A16. Dates of Technical Performance: Rabbit Arrival Date Dosage Period [Days 7 through 20 of presumed gestation (DGs 7 through 20)] Toxicokinetic Sample Collection (DG 21) Caesarean-Sectioning Period (DG 29) 21AUG 98 23 AUG 98 - 09 SEP 98 06 SEP 98 - 10 SEP 98 14 SEP 98 -- 18 SEP 98 A17. Records Maintained: The original report, raw data and reserve samplesof 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 draft final report, after which time the Sponsor will decide their final disposition. All unused test article suspensions were discarded atthe Testing Facility. Unused bulk test article will remain at the Testing Facility until its disposition is decided by the Sponsor. B. Test Article Information: B.1. Description: PFOS -- an off-white powder B.2. LotNumber: 217 (Expiration date: May 2000) B.3. Date Received and Storage Conditions: The test article was received on 20 May 1998, and stored at room temperature. Prepared formulations were stored refrigerated. B.4. Special Handli ions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles or safety glasses and a face-shield) were taken when handling the bulk test article and prepared suspensions. B.5. Analysis of Purity: Information regarding the purity, identity, strength and composition of the test article is on file with the Sponsor. 000663 418-012.PAGE 114 C. Vehicle Information: C.1. Description: 0.5% Tween 80 prepared using 2% Tween 80, a clear viscous to yellow liquid, in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). C2. Lot Number: MO3HoS ca. ived and Storage Conditions: The 2% Tween 80 was received on 8 July 1998, and stored at room temperature. R.O. deionized water is available from a continuous source at the. Testing Facility and is maintained at room temperature. C.4. Special Handling Instructions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles or safety glasses and a face-shield) were taken when handing the vehicle. C.5. Analysisof 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. TestArticle Preparation: Suspensions of PFOS were prepared daily at concentrations of 0, 0.02, 0.2, 0.5 and 0.75 mg/mL. The test article was considered 100% pure for the purpose of dosage calculations. 000664 418-012:PAGE II-5 D1. SampleInformation: Sample Type _| Components Concenraton CStoonrdageens | snopodTo | oat smgpes HAUG ZAUG Se fgmsm][|=[f [= T[=E~y [faemer| Rese Aves rTfo J me fe Verice Reserve ll al =ll water temperate | Arches temperature | Archives 5. sDaumppilceaslwoesrameplreestwaarieantteahkdeenfToomseaFchaconc2cariBiarctaktuopyns.Onsatofsales wes ser for analysis: th remaining 5. FLasstddaayyooffpprreeppaarraattioino.n Homogeneity and stability of prepared formulations are on file with the Sponsor. D.2. Analytical Results: Concentration samples (2 mL) were taken on the first and last days of preparation for analyses by 3M Environmental Technology and Safety Services. The resultsof these analyses were not available at the time of this report. E. TestSystem: E.1. Species: Rabbit E2. Strain: New Zealand White [Hra:(NZW)SPF] E3. Supplier (Source): Covance Research Products, Inc. Denver, Pennsylvania Ed. Sex: Timed-pregnant female 000665 418-012:PAGE II-6 ES. Rationale for Test System: The New Zealand White [Hra:(NZW)SPF] rabbit was selected as the Test System because: 1) itis one non-rodent mammalian species accepted and widely used throughout the industry for nonclinical studies of developmental toxicity (embryo-fetal toxicityteratogenicity); 2) this strain of rabbit has been demonstrated to be sensitive to developmental toxins; 3) historical data and experience exist at the Testing Facility"; and 4) the test article is biologically active in the species and strain. E.6. TestSystemData: Number of Rabbits Approximate Date of Birth Approximate Age at Arrival Weight (kg) on DG 0 Weight (kg) at Arrival 133 21 MAR 98 5 months 277-420 284-421 E7. Method of Randomization: Upon arrival, rabbits were assigned to individual housing on the basis of computer-generated random units. Rabbits were assigned to one of five dosage groups (Groups | through V), 22 rabbits per dosage group, for the main portion of the study. An additional 19 satellite rabbits were assigned for toxicokinetic evaluation; five rabbits were assigned to each of the low and high dosage groups (Groups II and V) and three rabbits were assigned to each of the remaining dosage groups (Groups |, Ill and IV). Rabbits were assigned to dosage groups using a computer-generated (weight-ordered) randomization procedure based on body weights recorded by and at the Supplier (Covance Research Products, Inc.) on DG 0. ES. System of Identification: Cage tags were marked with the study number, permanent rabbit number, sex, test article identification and dosage level. Each rabbit was individually identified with a Monel self-piercing ear tag (Gey Band and Tag Co, Inc., No. MSPT 20103) inscribed with the rabbit's designated unique permanent number. 000666 418-012:PAGE Il-7 F. Husbandry: FA. Research Facility Registration: USDA Registration No. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 ot seq. F.2. StudyRooms: The study rooms were maintained under conditions of positive airflow relative to a10h0al%lwfareysahnadiritnhdaetpheanddebnteleynspuapsplsieeddtwhirtohuaghm9i9n.i9m7u%m HofEtPeAn cfihlatenrsge(sAirpoerClheoaurnof trhoeomsst)u.dy.RoRoomotmemtpeemrpaetruarteuraenwdahsumtiadrigteytewderate6m1onFittoor7e2dcFon(s1t6antCltyot2h2roCu)g;hout relative humidity was targeted at 30% to 70%. F.3. Housing: Rabbits were individually housed. All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratory Animals. F4. Lighting: Alinght:au1t2o-mhaotiucraslldya-rcko,nwtirtohlleeadcfhludoarrekscpeenrtioldighbtecgyicnlneinwgaast m1a9i0n0tahionuerdsaEtS1T2.-hours F.5. Sanitization: Cage pan liners were changed approximately three times each week. were changed approximately every other week. Cages F.6. Feed: Approximately 150 g of Certified Rabbit Chow #5322 (PMI Nutrition International, St. Louis, Missouri) was available to each rabbit each day until the first day of dosage, at which time approximately 180 g of the same certified feed winadsivoifdufaelresdtationeleascshstreaeblbi"tJe-taycphe"dafye.edeTrhseactetraticfhieedd tfoeeedacwhascaagvea.ilable from 000667 418-012PAGE II-8 F.7. FeedAnalysis: Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration for certified feed or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any agent present in the feed that was known to interfere with the results of this study. FS. Water: Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rabbits ad libitum from individual water bottles and/or from an automatic watering system (individual sipper tubes). Chlorine was added to the processed water as a bacteriostat. FS. Water Analysis: The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, Pennsylvania) and monthly for possible bacterial contamination (Analytical Laboratories, Inc., Chalfont, Pennsylvania). Copies of the results of the water analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any agent present in the water that was known to interfere with the results of this study. 000668 418-012:PAGE II-9 G. Methods: GA. Dosage Administration: OGuospe|| MRuabrbersr ||moohogssisoea|n| Gomrcaemtm)ion | Dosage voume 0 [an] same | To=rrTT oTo oa[ [ m 5 5e Jovwoosms [[oommeme]] [CCov[[moee[[ooweT ToaToToowm n[[vvoorreess]] To Tow Ton | ova [wow] 2TheRatemtoasrc4osgwnaessc1on0sihderesdc10o0%otpureevfoaratheonpurpose of dosage calclatons. G.2. Rationale for Dosage Selection: Dosages were selected on the basis of a dosage-range study (Argus Research Laboratories, Inc., Protocol 418-012P) G.3. RAoudtmeionfistration: Oral (stomach tube) G.4. Rationale for Route of Administration: The oral (stomach tube) route was selected for use because: 1) in comparison with the dietary route, is one of the possible the exact dosage can be accurately routes of human exposure. administered; and 2) it G.5. Freq ofu Adme inin strc atiy on: Appropriate dosages of the test article were tube) once daily to naturally-bred rabbits on administered orally DGs 7 through 20. (via stomach Dosages of 0 (Vehicle), 0.1, 1.0, 2.5 and 3.75 mg/kg/day of the test article were administered at a dosage volume of 5 weights recorded before ml/kg, adjusted intubation. The daily on the rabbits were basis of the individual body intubated at approximately the same time each day. G6. Length of Study: Approximately 4 weeks 0C0E69 418-012.PAGE 11-10 G.7. Method of Study Performance: The female rabbits were naturally bred by breeder male rabbits of the same source and strain before shipment to the Testing Facility. The rabbits were mated on five consecutive days and shipped to the Testing Facility on the day after the last day of mating. The day of mating was considered to be DG 0. A computer-generated (weight-ordered) randomization procedure was used to assign the rabbits to five dosage groups based on body weights recorded on DG 0 and supplied by Covance Research Products, Inc. All rabbits were observedforviability at least twice each day of the study and for general appearance several times during acclimation and on DG 0. Additional examinations for clinical observations of effects of the test article, abortions, premature deliveries and deaths were made at least once during the predosage period, once before each daily intubation and approximately 60 minutes after intubation during the dosage period. These observations were also made once daily during the postdosage period (DGs 21 through 29). Body weights were recorded on DGs 0 and 6 through 29. Feed consumption values were recorded daily throughout the study. G.8. Gross Necropsy: G.8.a. Satellite Rabbits Assigned to Toxicokinetic Sample Collection: On day 21 of presumed gestation (the day following the last dosage), toxicokinetic samples were collected from the rabbits assigned to the toxicokinetic evaluation. Following anesthesia of pentobarbital, blood samples (approximately 4 mL per rabbit) were collected from the inferior vena cava into serum separator tubes and centrifuged. The resulting serum (approximately 2 mL) was immediately frozen on dry ice and maintained frozen (-70C) until shipment to the Sponsor for analysis. The liver was excised, weighed, and a sample was taken from the right lateral lobe, frozen and retained at 70C until shipment to the Sponsor for analysis. Rabbits were Caesarean-sectioned and fetuses were examined grossly to the furthest extent. Fetuses and placentae were pooled per litter and retained frozen (-70C) until shipment to the Sponsor for analysis. After completion of sample collection, serum, liver sections, fetal and placental samples were shipped (frozen on dry ice) to 3M Environmental Technology and Safety Services, St. Paul, Minnesota. 000670 418-012:PAGE II-11 G.8.b. ScheduledSacrifice: All surviving rabbits were sacrificed by intravenous administration of Beuthanasiae-D Special euthanasia solution on DG 29. The thoracic, abdominal and pelvic viscera of each rabbit were examined for gross lesions. Gross lesions were preserved in neutral buffered 10% formalin for possible future evaluation (with the exception of parovarian cysts, which are common, spontaneous lesions in rabbits); all other tissues were discarded The number of corpora lutea in each ovary was recorded. The uterus was excised and examined for pregnancy, number and distribution of implantations, early and late resorptions and live and dead fetuses. Uteri from does that appeared nonpregnant were stained with 10% ammonium sulfide to confirm the absence of implantation sites. An early resorption was defined as one in which organogenesis was not grossly evident. A late resorption was defined as one in `which the occurrence of organogenesis was grossly evident. A live fetus was defined as a term fetus that responded to mechanical stimuli. Nonresponding term fetuses are considered to be dead (there were no dead fetuses). Dead fetuses and late resorption are differentiated by the degree of autolysis present; marked to extreme autolysis indicated that the fetus was a late resorption. Each Caesarean-delivered fetus was weighed, examined for gross external alterations and individually identified with a tag noting study number, litter number, and uterine distribution. Live fetuses were sacrificed by an intraperitoneal injection of Beuthanasiae-D Special. All fetuses were examined internally to identify sex and visceral alterations; cavitated organs, including the brain, were evaluated by dissection"; and the brain was free-hand crosssectioned (a single cross-section was made between the parietal and the frontal bones) and examined in situ. Fetal gross lesions were preserved in neutral buffered 10% formalin for possible future evaluation. Al fetuses were eviscerated, stained with alizarin red S"" and evaluated for skeletal alterations. Al skeletal preparations were stored in 80% glycerin with thymol crystals added to retard fungal growth. Late resorptions were examined 10 the extent possible. Representative photographs of fetal alterations are available in the raw data. Rabbits that aborted were examined on the day the observation was made. Pregnancy status and uterine contents were recorded. Aborted fetuses were examined to the extent possible, using the same methods described for fetuses. Uteriofapparently nonpregnant does were stained with 10% ammonium sulfide to confirm the absence of implantation sites. 000671 418-012:PAGE Il-12 G.9. StatisticalAnalyses: The following schematic represents the statistical analyses of the data: Type of Test I. Parametric Il. Nonparametric A. Bartlett's Test A. Kruskal-Wallis Test (75% ties) Significant at p<0.05 Norpdrametc Not Significant | Analysis of Variance Significant at p0.05 | Dunn's Test Not Significant Significant at p<0.05 Not Significant B. Fisher's Exact Test (>75% ties) odo Test Ill. Test for Proportion Data Variance Test for Homogeneity of the Binomial Distribution a. Statistically significant probabilfies are reported as either at p<0.05 or p<0.01 b. Used only to analyze data with homogeneity of variance. . Proportion data are not included in this category. d. Test for homogeneity of variance. 000672 418-012:PAGE I1-13 Clinical Test for observation and Homogeneity of other proportion data were analyzed the Binomial Distribution, using the Variance `Ccoonntsiunmupotuisondavtaalu(ees.g.a,ndmaltiettrenraalvebroadgyewsefiogrhtpse,rcbeondtymwaelieghftetcuhseasn,gepse,rcfeenetd resorbed conceptuses, fetal body weights, ossification site data) were analyzed using fetal anomaly Bartlett's Test data and fetal of Homogeneity of TVeasrtiawnacsenso"t asnigdnitfhiecaAnntal(>y0s.i0s5)o]f.VarIfitahnecAen"a,lywshiesnofaVpparroiparnicaetewa[is..s,iBganritfliectatn'ts i(npd<i0v.i0d5u)a,l Dgurnonuepst.t'sIfTtehsetA"nawlayssisusoefdVatoriiadnencteifwyatshensottataipsptricoaplrisaigtneiffiiec.a,ncBaeortflettth'es Test was significant (p<0.05)], the Kruskal-Wallis Test" was used, when less Ftihsahnero'rseEqxuaaclttTo e7s5t%"twieasswuesreed.preIsnecnta;sewsheinnwmhiocrhetthheanKr7us5k%alt-iWeaslwliesreTepsrtesweanst., ustsaetidsttiocaildleynstiigfynitfhiecansttat(ips<t0i.ca0l5)s,igDnuinfnic'asncMeeotfhothdeoifndMiuvlitdiupalle gCrooumpps.arisons was Count data procedures obtained at Caesarean-sectioning were evaluated previously described for the Kruskal-Wallis Test". using the Group | rabbit 8481 had a litter consisting of ten dead fetuses. occurrences can abnormally skew the distributionof the data, Because such statistical analyses `were were made with and excluded from without the values summarization and for this rabbit and litter. Data for statistical analyses; values are this litter presented on the individual tables. 000673 418-012:PAGE Il-1 Wl. RESULTS A. Mortality, Abortions, Clinical and Necropsy Observations (Summary Tab1l; e Individual Data -Tables 3, 14 and 15) A Abortions Aintotthael3o.f75temngr/akbgb/itdsayabdoorsteadg;eognreoupin. the 2.5 mg/kg/day These abortions dosage group occurred after and the nine" completion of 24,25 or 28)] the and dosing period [on were considered days 22, 24, 25 or 28 of related to the test article gestation because (DGs they 22, occurred at dosage-dependent incidences in other rabbits survived to scheduled sacrifice. the two highest dosage groups. All 2.5 mglkgiday wDeoree8n5o17adavbeorrsteedcleiingichatlloabtseerrevsaotripotnisonbseofonreDGabo2r5tinagn.d wTahisssdaocreifliocsetd.boTdhyewreeight eaingdhtfeiemdplcaontnastuimopntsiiotneswaansd salelvetrieslsuyerseadpupceeadraefdtenroDrmGal.13. AuNteoclyrsoipssyprreecvleuadleedd further evaluation of the eight late resorptions. 3.75 malkalday aDdoveer8s5e34clianbiocarlteodbsoenrevadteiaondsfbeetufsoroenabDoGrti2n2g.anTdhiwsadsoseacgreinfiecreadl.lyTlhoestrewewiegrhte annod hreavdeasleevderseelvyernedliuvceefdetfueseedscionntshuemuptteirouns vaanldueaslltthirsosuuegshoauptptehaerestdudnyo.rmaNle.crAolplsy ftiestsuuseesexaapmpienaarteiodn.norFmeatlusfeorst1h,e2ir, d3e,v4e,l5o,p6me.n7taalndag8ehsaadt ngortosossseifxiteedrnpaulbaensdastoft skeletal examination. `Daodeve8rs5e37clianbiocarlteodbsfeirvvealtaitoenrebseofroprteioanbsorotnioDnGwa2s5 aabnsdenwtafsescaecsri(fiDcGed.24).ThTehoinsldyoe gtehnreoruaglhloyutlotsthewesitugdhyt. anNdechraodpssyevreerveelaylrededfuicveedimfpeleadntcaotniosnumspitteisoannvdalaluletsissues appeared normal. resorptions. Autolysis precluded further evaluation of the five late Dweoree8n5o38adavbeorrsteedcleiingichatlloabtseerrevsaotripotnisonbsefoonreDGabo2r4tinagn.d wTahisssdacoreifgiecende.ralTlhyelroest wNeeicgrhotpsayndrehvaedalseedvenrienley irmepdlauncteadtifoenedsicteosnsaunmdptalilotnisvsauleuseasptpheraoruegdhonuotrmtahle. stOudnye. conceptus was of the eight late presumed cannibalized resorption. and autolysis precluded further evaluation ** Significantly different from the vehicle control group value (p<0.01). 000674 418-012:PAGE lll2 Doe 8539 aborted seven live fetuses on DG 24 and was sacrificed. The only adverse clinical observation before abortion was scant feces (DGs 22 and 23) This doe generally lost weight and had severely reduced feed consumption values throughout the study. Necropsy revealed seven implantation sites and all tissues appeared normal. All fetuses appeared normal for their developmental ages at had not gross external ossified pubes and and soft tissue fetuses 2 examination. and 3 had not Fetuses ossified 1,2, 3,4, 5, 6 and sternal centra at 7 skeletal examination nDooead8v5e4r0seabcolirntiecadlfoobusredrevaatdiofnestubseefsooren aDboGrt2i5nga.ndThwiassdosaecrliofsitcewde.ighTthearnedwfeereed cthorneseumlipvteifoentuvsaesl,ueosnweedreeasdevfeerteulsyesraendudcseidx alfatteerrDesGor1p5t.ionNsecinrothpesyutreervuesa.leAdll tissues appeared normal at necropsy. All fetuses appeared normal for their developmental ages at gross external precluded further evaluationof the six and late soft tissue examination. resorptions. Fetuses 1, Autolysis 2, 3,5, 7, 8,9, and and 10 had not ossified pubes and fetus 2 short maxillae at skeletal examination. had incompletely ossified nasal bones. Doe 8542 aborted six dead fetuses on DG 28 and was sacrificed. The only adverse clinical observation before abortion was scant feces (DGs 21 to 23 and 27). This doe lost weight and had severely reduced feed consumption after DG 18. normal. NTewcorocposnycerpetvuesaleesdweeirgehtpirmepsluanmteadticoannsniitbeasliaznedd aallntdisalsluseisxaapbpoeratreedd fetuses were partially cannibalized. examination. All fetuses appeared normal at soft tissue aDdoveer8s5e44clianbiocarlteodbsneirnveatdieoandbfeeftourseesaboonrtiDoGn 25 and was sacrificed. was scant feces (DGs The 19 to only 20 and 220 24). consumption After DG 12, until sacrifice. this doe lost weight Necropsy revealed and nine had severely implantation reduced sites and feed all ttiissssuueesexaapmpienaatrieodn.norFmeatlu.seAlsl1f,e2t,us3e,s4a,p5,pe6a,r7e,d8naonrdma9l haatdgrnoostsoesxstifeirendalpuabndessoft and fetus 3 had not ossified sternal centra at skeletal examination. aDdoveer8s5e47clianbiocarlteodbseeirgvhattliaotnesrbeesofroprteiaobnosrotnionDGwe2r2e asncadnwtafsecseascr(ifDiGced1.8)Tahned oanlryed substance in the cage were reduced DGs 11 p10a2n1.(DNGe2c1r)o.psByodryevweeailgehdtesigahntdifmepeldanctaotnisounmpsittieosnanvadlualels tissues appeared normal. resorption. Autolysis precluded evaluation of the eight late Doe 8548 aborted one dead adverse clinical observation fetus on DG 25 before abortion and was was sacrificed. The scant feces (DGs 21 only and 23 to 24). This doe lost reduced after DG 17. weight after DG 15 and feed consumption was severely Necropsy revealed seven live fetuses, one dead fetus and 000675 418-012:PAGE Ill-3 two late fetuses resorptions in the uterus. appeared normal for their Al tissues appeared developmental ages normal at gross at necropsy. extemal and All soft tissue examination. The late resorptions appeared examination; autolysis precluded further evaluation. normal at Fetuses gross external 1,2, 3,4, 5,6, 7, 8 and 11 had not ossified pubes at skeletal examination. A.2. ClinicalObservations oIbnscerrevaasteidonnsumofbesrcasnotfodroneosfienctehse; 3t.h7e5imncgi/dkegn/cdeayofdscoasnatgefegcreosupwahsadsignificant (p<0.01). Scant feces also occurred in one and three rabbits in the 1.0 and 2ra.b5bimtglalksgoldhaayddsoosftaogrelgirquoiudpfsecreesspeocntiovenley.dayT.heOn1.e0 dmoge/kign/tdhaey3d.o7s5amgge/kggr/oduapy dosage group abortion. had red substance in the cage pan associated with impending All other adverse clinical observations, and any statistically significant (p<0.01) increases in the incidences the test article because the of these observations, were considered incidences were not dosage-dependent. unrelated These to observations included ungroomed coat. localized alopecia on the limbs, back and/or underside and A.3. NecropsyObservations With the exception of persistent adverse lesions were identified at necropsy. clinical observations, no additional gross B. Maternal Body Weights and Body Weight Changes `Summaries - Tables 3 and 4; Individual Data - Table (Figure 16) 1; M1.a0t,e2m.a5lanbdod3y.7w5eimggh/tkgga/idnasywdeorseagreedgurcoeudpsoratbomdoystwetiagbhutlaltoesdseisntoecrvcaulrsrdeudriinngthtehe dosage period; these reductions or body weight losses were statistically significant (p<0.05 1013, 1310 16 and or p<0.01) 16 10 19. in the 3.75 mg/kg/day dosage group on DGs 10 During the first three days of the gains continued to be reduced in postdosage the 1.0 and period (DGs 21 to 24), 2.5 mg/kg/day dosage body weight groups; significant (p<0.05) body weight loss occurred in the 3.75 mg/kg/day dosage sgirgonuipficdaunrtinbgodthyiswepiergihotd.redAusctaiornessuoltr obfotdhyewseeirgehdtucltoisosness, (dpo<s0a.g0e5-odreppe<n0d.e01n)t, opecrciuordre(dcailnctuhlaete1.d0,as2.D5Gasn7d 3t.o7251)m.g/kg/day dosage groups for the entire dosage Dosage-dependent reductions 3.75 mg/kg/day dosage groups in body for the weight gains occurred entire gestation period in the (DGs 2.5 0 to and 29) and for the gestation period after the initiation of dosing (DGs 7 to 29; significant at 000676 418-012:PAGE lll-4 Pps0.01 in the 2.5 mg/kg/day dosage group). Average body weights were s3i.g7n5ifmigca/nktgl/y draeydudcoesdag(ep<g0r.o0u5po,rapss0c.o0m1p)aroendDwGisth 1t7hethvreohuigclhe2c4onitnrotlhegroup values. Body weights and dosage of PFOS. body weight gains were unaffected by the The significant reduction (p<0.05) in body 0.1 mg/kg/day weight gain in the 0.1 mg/kg/day dosage related because it was group on transient DGs 10 and not dtoos1a3gwe-adsepnoetndceonnstidered treatment- C. MVaatleurensal(SAubmsmolaurtiees(g-/Tdaayb)leasn5daRnedla6t;ivIend(iav/ikdgu/adlay)DatFaee-dTaCbolnes1u7m) ption Adbursionlguttehe(gd/odsaay)geanpderrieoldat(iavfete(rg/DkGg/d1a0y))ifnetehdec2o.n5saunmdpt3i.o7n5 vmagl/ukegs/wdeayredorseadguec.ed groups; both of tthheesseegrreoduupcstioonnsDwGesre13sttatois1t6ic(arlleylastiigvneiofnilcya)n,t (p<0.05 16 to 19 or p<0.01) and 19 to in 21 one or Atbwsooglruotuepasnddurrienlagtitvheefpeoesdtcdoonssaugmeppteiroinodv;atlhueesrecdouncttiinounesdwteorbeegreneedruacleldy in these statistically significant (p<0.05 or p<0.01) on DGs 21 to 24. Rveafllueecstiwnegrtehreesdeuecfefdectosr osifgtnhiefitceasnttlayrtriecldeu,caebdso(lpu<t0e.0a5nodrrpe<l0at.i0v1e)feinedthceo2n.s5umapntdion 3.75 mg/kg/day dosage groups for the entire period after the initiation the entire of dosage dosage (DGs 7 period to 29). (DGs 7 to 21), and Absolute and relative feed consumption 1.0 mg/kg/day dosage of PFOS. values were unaffected by the D. aCnades8a;reIandni-vSiedcutalioDnaitnag -aTnadblLeitste1r8Otbhsreoruvgahti2o0)ns (Summaries - Tables 7 2P1re(g9n5a.n4c%y) orcacbbuirtrsediniena2c0h(d9o0s.9a%g)e,g1r9ou(p.86.C4a%)e,sa1r9ea(n8-6s.e4c%t)i,on1i7n(g7o7b.s3e%r)vaatnidons on DG 29 were based respective dosage on 20, groups. 18, 19, 16 One and and nine 12 pregnant rabbits does aborted in the in each 2.5 and of the five 3d.o7s5agmeg/gkrgo/udpaydodeos(a8g48e1g)rtohuapts,hardeaspelcitttievrecloyn.siVsatliungesoffotrenondeea0d.1fmetgu/skegs/dwaeyre excluded from considered to data be a tabulation and non test article statistical analyses; this total related, spontaneous event ltter death was (Fpet<a0l.0b5odayndwepi<g0h.t0s1,(troetaslp,ecmtailveelay)ndinftehmeal2e.)5 waenrde3s.i7g5nimfgic/akngtl/ydaryedduocseadge groups, abisolcoogimcpaallryeidmtpoortthaenvteohricslteaticsotnitcraolllygsriogunipfivcaalnutesd.iffTehreernecewseirnethneoloittthereraverages fwoerrceornpoordaoelsutweia,thiamlpllcaonntacteipotnu,selsiverefestoursbeesdaanndd eaallrlpylaocrelnattaeereaspoprpetairoensd. noTrhmaelr.e 000677 418-012:PAGE Ill5 E. Fetal Alterations (Summaries -Tables 9through 13; Individual Data - Table21) oFectcaulraalttelroawtiionncsiwdeenrceesdeifnitnheids assp:ec1i)esmaalnfdorsmtaratiino);nsan(idrr2e)vevrasriibalteiocnhsa(ngceosmmthoant findings in this development). sLpietctieresa/vsterraaigne,sawnedrreevcearlsciubllaeteddelfaoyrsspoerciafciccelfeetraaltiososnisfiicnation sites as part of the evaluation of the degreeoffetal ossification. Fetal evaluations were based on 175, delivered live fetuses in 20, 18, 19, 16 162, and 152, 130 12 litters and 108 DG 29 Caesarean in the 0 (Vehicle), 0.1, 1.0, 2.5 for garnodss3.ex7t5ermnga/lk,gs/odfatytidsossueagaendgrsokueplse,tarleaslptecetriavteiloyn.s All and fetuses were examined fetal ossification site averages. in the 2.5 lt was also possible to examine the ten dead fetuses from mg/kg/day dosage group for gross external, soft tissue and litter 8481 skeletal alterations. substance. The embryonic sacs of all of these fetuses contained a dark red One fetus had downward flexion of the forelimbs, an observation faestsuoscihaatdedanwgiutlhaitneudtheyrooicdoamlparee,sasnidontw(sokefleettuasleossshiafdicoantlioyntneontoarfsfiexctceadu);daalnother vertebrae. The remaining six fetuses appeared normal. EA. SummaryofFetal Alterations (Summary -Table 10; Individual Data Table 22 fCoormfbeitnalatalitoenratoifonmsa.lfoIrn mtahteio0n(sVeahnidclvea)r,ia0.t1i,on1s.0r,es2u.l5teadndin 3t.h7e5fmogll/okwgi/ndgaiyncdiodseangcees g8r(o6up6s.,7%r)eslpietcetrsivhealyd,f1e4tu(s7e0s.0w%it)h, o1n1e(o6r1.m1o%r),e 9al(t4e7ra.t3i%o)n,s 4ob(s2e5r.v0e%d). aInndthese `alstaemreatrieosnpsewcetirvee2d5os(a14g.e3%g)r,ou2p5s,(1t5h.e4t%o)t,al1n4u(m9b.e2r%s)o.f5fe(t3u.s8e%s)"wiatnhdan19y i(d1e7n.t6i%fi)e.d One or 17.4% more of the afletteursaetsiopnesrolcicttuerrrien dthien afivveerraegsepsecotfiv1e4.d1o%s,ag1e7.g0r%o,up9s..5%T,h3e.6% and significant alterations reduction in the 2.5 (p<0.05) in mg/kg/day the total dosage number of group was fetuses with identified not considered treatment- related because the expected rather than a decrease, in the response to a toxicant number of alterations. would be an increase, oDrelpa<y0s.0i1n)fefteatlalosbsoidfyicawteiiognhatsssioncithaete2d.5wiatnhdth3e.7s5igmnigf/ikcgan/tdlayyreddouscaegde(gpr<o0u.p0s5 were evident ossified as significant reductions (p<0.05 or sternal centers in the 2.5 and 3.75 p<0.01) in mg/kg/day the litter averages for dosage groups and significant reductions and metacarpals and (p<0.05 or significant p<0.01) in reductions the litter (p<0.01) averages for ossified in the litter and fetal hyoid incidences of incompletely ossified pubes in the 3.75 mg/kg/day dosage group. "+ Significantly different from the vehicle control group value (p<0.01). 000678 418-012:PAGE Il6 All other fetal gross external, soft tissue and skeletal alterations (malformations and variations) were considered unrelated to the test article because: 1) the. incidences were not dosage-dependent; and/or 2) the incidences were within ranges observed historically at the Testing Facility". E.2. Fetal Gross External Data - Table 21) Alterations (Summary - Table 10; Individual E2.a. Malformations `The fist 8448-1. dSikgeiltewtaalseaxbasmeinntatoinonbootfhthfiosrefleitmubssrfeovrevaelheidclaebsceonnttro1lstgrmeoduipafletaunsd distal phalanges and 1st metacarpal on both forelimbs. occurred in this fetus. No additional alterations E.2b. Variations One fetus (8473-6) in a 0.1 mg/kg/day dosage group (8543-3) in a 3.75 mg/kg/day dosage group litter had litter and another downward flexed fetus forepaws, afentuosbs8e5r4v3a-t3iornevaesasloecdiaatevdarwiiatthioinn iuntsekruollcoomsspirfeiscsaitoionn. (hSokleeleintatlheexpaamriientaatliboonneo)f. Fetus 8473-6 had no additional alterations. EJ. Fetal Soft Tissue Alterations (Summary - Table 11; Individual Data Table 21 E..a. Malformations No soft tissue malformations were observed. E3.b. Variations E3bA. Eyes Olenfteeyceo,ntarovlardioastaiogneggerneoruapllfyetautstr(i8b4u4t9a-b6le) thoatdraaucmiarcduumrcionrgnperaolcehsesminogr.rhaSkgeeoleftatlhe `evaluation of this fetus (regular nasal-frontal revealed variations in suture and angulated ossification alae). of the skull and hyoid a. See APPENDIXE (HISTORICAL CONTROL DATA). 000679 418-012:PAGE IIl-7 E3.b.2. Lungs The intermediate lobe of the lungs was absent in 2, 7**, 1, 0 and 1 fetuses from 2,4,1,0.and1 litters in the 0 (Vehicle), 0.1, 1.0, 2.5 ad 3.75 mg/kg/day dosage groups, respectively. Two of the 0.1 mg/kg/day dosage group fetuses (8476-8, 8483-4) also had fused 3rd and 4th sternal centra. The significant increase (p<0.01) in the fetal incidenceofthis observation in the 0b.e1camugs/ek:g/1d)aythidsovsaargiaetigornouinp was lung not considered development is related to the test article frequent in this rabbit strain(', 2) the fetal and litter incidences are within the historical rangesof the Testing fFoacritlihtey,un3i)t iotfwmaesansoutredmoesangte,-tdheepelintdteernt; and 4) it was not significantly increased E.4. Fetal Skeletal Alterations (Summaries - Tables 12 and 13; Individual Data - Table 21 E.4.a. Malformations E.4.a.1.ThoracicVertebrae/Ribs Interrelated vertebral/rib malformations or malformations of only the ribs occurred in one, three and one fetuses in the 0 (Vehicle), 0.1 and 1.0 mg/kg/day dosage groups, respectively. These types of vertebral/rib malformations are relatively common at maternally toxic dosages in rabbits and generally considered to be secondary to maternal stress". F3redtutsho8r4a5c2ic-1veirnttehberaveahnidclae scpolnittr2olndgrloefutpribh.adThaislefftethuesmiavlesortheabdraapsryemsmeenttraisc t1hste 10 3rd sternal centra and an angulated hyoid ala. Fetus 8476-1 in the 0.1 mg/kg/day dosage group had fused centra of the 11th and 12th thoracic fveetrutseb8r4a7e6-a7n,dhaadbiffiudsceedn5ttrhumanodf t6hteh r1i2btshatshotrhaeciocnlvyeratletebrraat,ioan.nd Fietstulistt8e4r8ma5t-e2, in the 0.1 mg/kg/day dosage vertebra, fused left centra ogfrtohuep 1h1atdh aa nsdma1l2lthleftthaorracchicofvtehrete1b1rtahe tahnodracpircoximate. bases of the 10th and 11th left ribs. Fetus 8500-3 in the 1.0 mg/kg/day dosage group had split 8th right and 7th left ribs as the only alteration. E.4.a.2. Lumbar Vertebrae Fetus 8500-8 had aleft hemivertebra present as the 1st lumbar vertebra and a bifid centrum of the 2nd lumbar vertebra. .- Significantly different from the vehicle control group value (p<0.01). 000680 418-012:PAGE Ill-8 E.4.2.3. Metacarpals/Phalanges The 1st medial and distal phalanges and 1st metacarpal on both forelimbs were absent for vehicle control group fetus 8448-1 that had missing first digits at external examination. E.4b. Variations E4b.A. Skull oCsosimfimcoatniosnmsailtlesirwrietghuilnartithieessuitnuorsessifoircactailovnaroifath(enassaklu,lflr"on[ttahleoprrpeasreientcael of small bones) and/or irregular shapingor fusion of the bones] occurred in 7, 9, 3, 1 and 10 fetuses in 6, 7, 3, 1 and 3 litters in the 0 (Vehicle), 0.1, 1.0, 2.5 and 3.75 mglkglday dosage groups, respectively. Irregular ossificationof the nasal bones (irregular nasal-frontal suture, fused, intemasal, midline suture displaced and intranasal) were the most common of these small irregularities in skull ossification, occurring in 6, 9, 3, 1 and 1 fetuses in 5, 7, 3, 1 and 1 litters in the five respective dosage groups. The significant reduction (p<0.05) in the fetal incidenceof variations in skull ossification in the 2.5 mg/kg/day dosage group was considered unrelated to the test article because the expected effect of a toxicant would be an increase in delayed skull ossification, rather than a decrease. The significant increase. (p<0.01) in the fetal incidence of variations in skull ossification in the 3.75 mg/kg/day dosage group was considered to reflect the significantly increased (ps0.01) fetal incidence of holes) in the parietal and unrelated to the test article, as discussed below. The fetal incidence of nasal midline suture displaced was significantly increased (p<0.01) in the 0.1 mg/kg/day dosage group but was considered unrelated to the test article because: 1) it was not dosage-dependent; and 2) the litter incidence was not significantly increased. The fetal incidence oaf hole in the parietal was significantly increased (p<0.01) in the 3.75 mg/kg/day dosage group. This increase was considered unrelated to the test article because the observation occurred in seven fetuses from only one high dosage group liter (8543-1, -3, -4, -5, -6, -8 and -10), so the litter incidence, the more liter also relevant had not parameter", was ossified pubes; all not significant. Two of the fetuses in this other fetuses had no additional alterations One 0.1 mg/kg/day dosage group fetus (8471-1) had unossified premaxillae as the only alteration. + Significantly different from the vehicle control group value (ps0.05) + Significantly different from the vehicle control group value (p<0.01) 000681 418-012:PAGE Ill-9 E4b.2. Hyoid One or both alae of the hyoid were angulated in 10, 4, 7, 3 and 5 fetuses in 6, 3, 3,2:and 3litters in the 0 (Vehicle), 0.1, 1.0, 2.5 and 3.75 mg/kg/day dosage groups, respectively. One fetus (8452-1) in the vehicle control group also had vertebrallrib malformations and asymmetric 1st to 3rd stemal centra, as previously described. E.4.b.3. Vertebrae E4b3.a. Cervical One 0.1 mg/kg/day dosage group fetus had unilateral ossification of the centrum of the 2nd cervical vertebrae as the only alteration E.4b3.b. Thoracic One vehicle control group fetus had unilateral ossificationof the centrum of the 5th thoracic vertebrae as the only alteration. E4b3.c. Lumbar Bifid centrum of a lumbar vertebra occurred in one fetus in the 1.0 and d3.o7s5agmeg/gkrgo/udpayaldsooshaagdeagrhoeumpisv,errtesepbercatipverleys.entFeatsuas 8l5u0m0b-a8r vinerttheebr1a.,0amsg/kg/day previously described. The fetus in the 3.75 mg/kg/day dosage group had no additional alterations. E4b3.d. Caudal A misaligned 17th the only alteration. caudal vertebra occurred in one vehicle control group fetus as E.4bA. Ribs Cervical ribs were present at the 7th cervical vertebra for one fetus in each of the 0 in (Vehicle) and these fetuses. 2.5 mg/kg/day dosage groups. No additional alterations occurred Thickened areas (8456-4) and one in two or three 1.0 mg/kg/day ribs occurred in one dosage group fetus vehicle control group (8493-7). No other fetus alterations occurred in these fetuses. 000682 418-012:PAGE IIl-10 E405. Sternum Fused 3rd and 3 fetuses from d4itfhfesrteentmalilttceersntirnathoecc0ur(rVeehdicalse)thaenodnl0y.1skmegl/ektagl/dalatyerdaotisoangien 2 and groups, respectively. Fetus centra. 8452-1 This in the vehicle fetus also had vceorntterborlalg/rroiubpmahlafdoramsaytmimoentsraincd1satntoan3grudlastteedahlyoid ala, as previously described. E.4bs6. Pelvis oFsosiufiredfeptuubseess. frTowmotwoof*t*he3s.e75femtgu/skegs/adlasyodhoasdagaevagrrioatuiponliitntersskuhlaldosisnicfoicmaptlieotnely t(hhoelfeetianlpaarnidetlailt)t,eraisncdiedsecnrciebsedofptrheivsiooubssley.rvaTthioenswigansifcicoannstidinecrreedasaete(spt<0a.rt0i1c)lein related delay body weights in in ossification and to reflect this dosage group. the significantly reduced (p<0.01) fetal E.4.b.7. Fetal Ossification Site Averages T(ph<e0.a0v5eroargpe<0n.u0m1b)eirnstohfeo3s.si7f5icmagt/ikogn/sdiatyesdpoesragfeetugsrwoueprefosrighnyiofiidc,anstltyeraelducceentders saingdnifmiectaanctalrypraelsd.uceTdh(ep<a0v.e0r1a)geinntuhmeb2e.r5smogf/kosgs/idfaiyeddsotsearngael gcreonutpe,rsaswecroempalasro.ed with the vehicle control observed historically at group value. the Testing The values were below Facility. These delays in the ranges ossification were considered effects (p<0.05 or p<0.01) of the test fetal body article associated with the significantly weights in these dosage groups. reduced Tluhmebaarv,ersaacgrealnuamnbdecrasudoafl)o,ssriifbisc,atsitoenrsniutmes in the vertebrae (manubrium and (cervical, xiphoid), thoracic, forelimbs o(ccacruprarlesd aantdsipmhiallaranignecsi)deanncdeshiinndllititmebrss i(ntaalrlsadlos,samgeetagtraorusaplssaanndd dpidhanloatnges) significantly differ. F. Satellite Rabbits (Individual Data - Tables 14 through 20 Awlelrreabnboitasdivnetrhseescaltienlilciatle odbosseargveatgironosupisn tsuhrevsievdeodesto. scPhaetdteurlnesdosfacbroifdiycew.eiTghhetre tghaeinsaanmdefdeoesdacgoensleuvmelpst.ioOnnwleyroenceormapbabirtabinlethteo3t.h7e5rmabgb/iktgs/idnatyhdeomsaaignesgtruoduypat wliattser npoatrapmreetgenrasntwaetreCaceosmarpeaarna-bsleectaimooninnggtohne fDivGe d21o. saCgaeesgarroeuapns-.seTchteiroeniwnagsand not a reduction in fetal body weights at the 2.5 and 3.75 mg/kg/day dosages as. "+ Significantly different from the vehicle control group value (p<0.01). 000683 418-012PAGE Ill-11 wAvaesroabgseelrivveerdwfeoirgthhtesmfaorinprsetgundayntradbboietss.weAlrled1o4e0s.5ap+pe20a.r2e,d1n2o7.r1ma+l a15t.n2,ecropsy. 31.2735.9mg4/3k3g./2d,a1y1d5o.s+7ag5e.7graonudps9,0r.e7sp+e2ct2i.v2eliyn.thTeh0e(Vdeahtiaclfeo)r,ra0b.b1i,ts1.a0,ss2i.g5naenddto the satellite portion of the study are provided in individual tables only 000684 418-012:PAGE IIl-12 REFERENCES: 1. U.S. Food and Harmonisation; DGruuigdeAldimnieniosntrdaettieocnti(o1n99o4f)t.oxiIcnitteymtaotiroenparlodCuocntfieonrefnorce on medicinal No. 183. products. Federal Register, September 22, 1994, Viol. 59, 2. U.S. Food and Drug Administration. Good Regulations; Final Rule. 21 CFR Part 58. Laboratory Practice 3. JPraapcatniceeseStMainnidsatrrdy foofrHSeaafletthyaSntdudWieelsfoanreDr(1u9g9s7,).MHGoWoOdrLdaibnoarnacteorNyumber 21, March 26, 1997. 4. EounrtohpeeaacnceEpctoannocmeicbyCtohmemEuunriotpye(a1n98E9c).onoCmouincciClomdemcuinsiitonyoonf a2n8 OJuElCy D1989 decision/recommendation on compliance with principles of good laboratory practice. Legislation. 32(No. Official Journal of the European L 315; 28 October): 1-17. Communities: 5. Cshtruidsytioafn,thMe.St.e,raHtoogbeenrimcapno,teAntMia.l aofndsuSsmpietnhs,iTo.nHs.Fo.f (t1r9i8n2i)t.rofDluoosraengoen-erange (2(T1N)F:)40ad(m#i1n4i3)s.tered orally to New Zealand White rabbits. Toxicologist 6. Cnharlitsrteixaonn,eM.(SP.ro(c1e9e8d4i)n.gsReofprNoadlutcrteixvoenetoSxiycmiptyoasinudmt,erNateowloYgoyrekvaAlcuaatdieomnsyof of Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 7. Feussner, EL, Lightkep, G.E., Hennesy, RA., Hoberman, AM. and Christian, M.S. (1992). A decade of rabbit fertiity data: control animals. Teratology 46(4):349-365. Study of historical 8. IUnssteoitfuteLaobfoLraabtoorraytAonriymAanlsi.malNaRteisonoaulrcAecsad(1e9m96y).PreGsusi,dWeafsohritnhgetoCna,reD.aCn.d 9. Salewski, E. (1964). Implantationsstellen Farbemethode zum am Uterus der Ratte. mAarkcrho.skPoatphiosl.chEexnp.NaPchhawremiaskovlon 247:367. 10. Staples, fetuses. RE. (1974). Detection of Teratology 9(3)A37-38. visceral alterations in mammalian 11. tSetacphlneisq,ueREi.n thaendKOSHc-hanleilzla,rVi.nL.re(d1964)m.ethRoedfifnoermfeetnatl ibnornaep.idSctlaeianriTnegchnol. 39:61-63 000685 418-012:PAGE I-13 12. Snedecor, G.W. and Cochran, W.G. (1967). Variance test for homogeneityof the binomial distribution. Statistical Methods, 6th Edition, lowa State University Press, Ames, pp. 240-241. 13. Sokal, RR. and Rohlf, F.J. (1969). Bartlett's test of homogeneity of variances. Biometry, W.H. Freeman and Co., San Francisco, pp. 370-371. 14. Snedecor, GW. and Cochran, W.G. (1967). Analysis of Variance. Statistical Methods, Pp. 258-275. 6th Edition, lowa State University Press, Ames, 15. Dunnett, C.W. (1955). A multiple comparison procedure for comparing several treatments with a control. J. Amer. Stat. Assoc. 50:1096-1129. 16. Sokal, RR. and Rohlf, F.J. (1969). Kruskal-Wallis Test. Biometry, W.H. Freeman and Co., San Francisco, pp. 388-389. 17. SMiceGgrealw,-HS.il(l1,95N6)e.w YNoornkp,arppa.me9t6r-i10c4.Statisticsforthe Behavioral Sciences, 18. Dunn, O.J. (1964). Multiple comparisons using rank sums. Technometrics 6(3):241-252. 19. Christian, M.S., McCarty, R.J., Cox-Sica, D.K. and Cao, C.P. (1987). Recent increases in the incidences of skull, lung and rib alterations in vehicle control New Zealand White rabbits. J. Amer. Col. Toxicol. 6(4):562 (#126). 20. Khera, KS. (1985). Maternal toxicity: A possible etiological factor in embryo-fetal deaths and fetal malformations of rodent-rabbit species. Teratology 31:129-153. 000688 APPENDIX A REPORT FIGURE 000687 MATERNAL BODY WEIGHTS Figur1e - -| - rE mn EaeeAl oon || oF [| we 3 a pT AliaSN ETE TN -- = |: 0 oF 72nd FrFaagx FE 3& hed 4 H0 Bh Hw # B APPENDIX B REPORT TABLES 000689 es ee ZZTF. Tae gYiiooooufrhoEyn rsED Euds go |iE P ESmHEEmEoEBa mSlLih IiImAInRDea Sr Tm ---- IIEEEmEEEEEmngIIomLniaIioaImnEns 3g3 BR ETI, eneset grow tue tgp.ta. &2 SE AE ly | : gg 3 - Te a g z ] EEBH rey TUR TP ee : TE mER ; LR uri or EN 3 2 & g mL mm mm mee ew mew man 8Emure mmumso. mamsseom msmeom omaesees oe acsem ese een IN Immonen me eee eee FE a a ot FO rt tr ee z EEEEcE s & "+ Slaniticantly Sifferen from the venicle control group value (eo.on) 8 iz Senn me amie ween ee LOT marin men mien anew Lwin TREN Tha :S 8 oars 21 + 39 Wowie. 0700 oarsA ea 0s 200 ea1s 0sTO woEs I0a0 3 3 Wess ewsen meson wwien addin bia g VEER i LEEEE EE Se rm g omnis men meine eB DIDO NERD TL EE ie g z SON mn WILLD mn med ads e oars 7.3 aw. was a. fine, fons, $ DEEa T E ETaThE ou ymTin,T o somata se - 2 SH g3 5 I ERLE Si eg wr ea & g g Te INIT onn on Em he wl mle ele wlan owle eee en g Ts Aeon | a es i Em nA ARE EEL a ed oe or : g2 E CoIleDEIn CETemnaiotcscoet ysoorm resn & 3 g 3 2 mmm ha o22: i 32 . g: i: . : 2 EE cr emwmtonynsmoves srmis mine, omen ET SL EE SATIIrs vaten tren en :g :5 & 8 iromer se) 20m oon en o 00 2 Wpaogosnted Woon fie in wen o en 23 g TEERTTETYETT wm xm wm nam 2 &E LonTrEamEcEoonEpr na nm mmo nm nom 35& a EBHIB Hh, HI BMRB ? BEERS na mmm ug, wi ww es CEEERSTTm osm mm mmm mm nm s $ wCuEn mw oE pin E gin JOgCmSOEnwES A 3 : g mem CHTum. ouwmoonmoommouam 5 3g EEE UAT wie wes wan em : EmErme.e A Aw.ae 2w8a.e 2wa8e E wenL EenE. 8 ED Bre we sensing wes er sori ron soem F LLE L EEEERSESk Ea Sn --rm--a g3 3 ag5 * 2 a TER mmomns oma omy ms om . Lr WMEED wmiem mmioew mai oem EE EE a  "052 oar or mREsoNED OESTATION o - or g33 or o :zz 2s 8  3 8 . EE i i i & Sn J i o EERE oo . 2 : is Ts g$ F=S Ba EBEEE E . 3 eisai SE, ES rr TR fi SR ew Eom EERRmE : & g a g 3 3 &g 3 s pater te thaTAAL Chimica) obsecvatices table (Table 30) for extern] chaervations conteed at seceopa. \ o OF amar to he iativsaua) chinical chaervations table (Table 14) for external cbeervations contiteed st BecropRy. & e: g3 # g : 5 &  wo wn a ow mmm g3g wo IwnLL8 wn m semeeene --a :: oo BE PB BEEEERER oe BTETTRETTEas coven wee ot 1 sr avers omens swiss z g s & & li Eh i B - Bion AvEAAGES AWECALCULATED ETHTUE RODDHILAR.(ALE g 3 I s ASETaxhicions wi uFROEESTrirmohann ;2 2 *3 i3: s 47 oun #443 hed 8 Titer which consisted on 10 desd fetuses; values excluded from grow averages and statistical analyses. g BEEERE ERELT S 8 ni ae : 5 & & g iMO AVERAGESARECALCULATED ITUE ROWED HiLocAa (4) VALOR g g Ena ; $gi 5 : | Ewes i 3 Z z : | FEEERrTe i 3 i g f < BODY WEIGHT AVERAGES AXE CALCULATED WITH THE ROUNDED KILOGEAW (KG) VALGE. Sh JE i g 3 & & i if g i 3 3 f 2 5 38 3 8 i 2 8 z g i o 2 = 3 : ; i i g g 3 3 g & sg 4." Spilled feed preciuded the calculation of this valve. 3 83 25 * 8 JE h RRR REE A QF Eh SR SRI Le ie frm ore vn a aes mre : 3 & :3 8 2g i: | i 2 3  a & 1 ii 3 i i 2: 5:i z : J a 38 & i 8:5 :i3 < i3 . ; :i % : : i@ 5 3 g :3 i 2 <g ; g T | g " a & 3 Ll OBitiidiiiiiiiiibigiig te LLL A] ; 3 3 FE SS A SS SE SES SE SE A a iii itil ridls TE NTE TE tolTedens totus: vatons exciuses trom geo averages aod staistica amiyuen i `vosage awour 111 3.0 marka/oAY Co B - gg BBFEIIzwip:r:r1r11r1r:r 11s1s13w8a:1a+y3 T EBVlI l rr rortotoroztororiiozory g 3 a S8 mDiEiP isrt iia bis iii bis iiis ids ins ag g3 3 , BREET iin SE hateau dad E BF a i Brule 5 1% oo 8 irri 8 o 8 s 8 a 8 5 3 = cE ) BE TE mens 1 & & i [J SS HS Sirs SERA a a Gy Ml sips SEE[T4T sere A IBEL E E I E E ey g z3 & ER TTT g #01} sk oEmoax oc fon io f EEE4 1H g 3 2 o FE A IE gS 2mooIiT 5828 1I18ae Bf 1 ig HEE 8 & 33 3 s Bool Ss Hn ro Boom uN Nd * 3a g Fd FI Tous oun uu : 3 be 3 2 . gL0 tEEE oe bE g Fd or mar noms na nas n 00 2 E@oGoh GEE EE BEOf 1d g g g $F hemes SF CE 3 we 3 3 FREE i sou i 3 5 3i pr GE 3 fd you TEE -% Fi i a va * 1a rete Bn 8 3 g ER mrIehmh EbRr EhAE T still 3 wow a tn a a mgm a mm 2 2 z ossar cao wewici woo - ~ 8S 3 g 3a 23 & g al alte wh ah ele al ate ists als ate wo oe nh ah aks 0h aheh oe g ean PE Ihl dI R EE oh Ee ad ET pre a i 0Durch mentor * "ies aorSeto inkRECobeDBans nT /" TES sin or coxvix i: ii :i 3 RE CE Re SW rae huis oom bar sesoerrion + Lai artis LJ omens retioficooRveik ? 8 i52zg 8 :2: & a tee althdh 2 FEE Gh a ehhe ah S EWSI TAr CCaEkL haSeLE5 o HeEWaCy LSEG 3 a WHO 4 sors Ferran 8 cH g: 3 3 5 &  g 2f i Ln ha NNR RS En PE TS g 8g PO a ah ha a hahah uh Nw ree i osono mai meri + ve sesrion L-Smores bettie orcenit 3 5: s 8 a : i . a haltah haha -- Foomiohahddhaes g wae CEE 0 a aa a as n 2 i onde" aIeEe wrennkeabeHsSecsndShs rerofiGoi wm 2 - 25 : CE TIRE ) & }: ii i rR 7 i onl Them meres, ' i 4 gH Gs .oS HS s a fEEiE i s. ere 2 5 8 & 8 Wu NBER Gp SPRCINENS WITH ALTESATICHSWAGER OF SPRCINENS EANINED FE Bh or 8 ar em A g me me wr uv 3 ET ------ ----; i. Ee ne . Topper 3 . wg g F i g 3 we awe eo a or YE En oPfiar,"eb 2:  ART HA ois hi Flame | Boe LC --" 3 &  Tonos, eremaoinre ) a BL crieA dBesSM den 1 cere cs aioe 4 rk 160 btu. Saves sere che from 2 gomweonw 8 w- wEsE F BoYle. 33} EL ca. san ot fn i || la wen ue ll fF ve pm w Ba . ERE g er g3e s--ba--mmnm,ie NL AER OF SPECIMENS WITH ALTERATIONS NGS OFSRC IMENS BANINED g33 5 ol 2 2J . - WE OF SPRCINRRSVITAALTERWBAETOIF SVORCNINESRS ROSIN - Cy --, - fap or - 2 a woo ame one Woke -- oe' ve -- ne EE a- - sa ese om om :ESoow wee one om i a a z2 o Worm 2g we umn we marie I. A A A . ve me 5 g of | ] ws aan em "ns oe en g monn ~ rte yi EAL ALE grat wo HE ba. mona oe 5 : oo hcA 3 8 o mme I. Pa g Flaw me 8 . a fm B EE ein Tyees I re 307.1% of ghutatiat. M1 ines WPLITES VGLMAY z | S PSE H Li a TT en, -- oe -3 3 $n 33a tmoreed sigh lateEesooun pSayt24 iof BoHVLmALLson:SOPN retioT bed oO rtoerL vereeS tios. Gos adtitional g sg . Bon353 biressseven Aefcines cn day 30 of gestation. ALY foture ppestad sormal a gross external and ste tisnue i E . E E e NA ELRAETTv, et 3 8 | REET ro oo : 28 3 EscoFt ion E BET roms a Span 8 Tm 2Earwe ssemm nwnr orco "sr os 2 g J ee worm on os or common 3 Baran 2 E SE Ee0re 2 i : i & APPENDIX C PROTOCOL AND AMENDMENT 000795 418-012:PAGE C-1 .PRIMED]ICA -_-- Argus50R8esSehaercehrLOarbioera:tSoruilesdninAgc. TelTeoplhootanxe.: ((231155))44433-.68751807 PROTOCOL 418-012 SPONSOR'S STUDY NUMBER: 6295.10 STUDY TITLE: PURPOSE: TESTING FACILITY: DIREC SPONSOR: STUDY MONITOR: ALTERNATE STUDYMONITOR: iOnraRlab(bSittosmach Tube) Developmental Toxicity Study of PFOS `PTFheOSpuornpoNseeowfZtehaislasntdudWyhiistteo[dHertaec:t(NaZdWv)eSrPsFe]efpfreectssumofed prfeetgunsacnotnsfeeqmuaelnetrtaobbeixtpsoasnudredeovfetlhoepdmoeentfroofmthiempelmabntraytoioannd tHoarclmoosnuirseoedf TtrhiepahratritdepGaulaitdee.linTehisstasgteusdyCevaanlduaDteosf tIhCeH reproductive process in a nonrodent species. A8r0g5uSshReeesheyarDcrihveL,abBourialtdoirnigesA, Inc. THeolresphhaomn,e:Pen(n2s1y5l)va4n4i3a-871190044-1297 Telefax: (215) 443.8567 ARsasyomcioantde GD.irYeocrtokr, oPfh.RDe.s,eDarAcBhT 3M 3M TCoexnitceorloBguiyldSienrgvi2c2e0s-2E-02 St. Paul, Minnesota 55144-1000 TMealrevpihnoTn.e:Case(6,51D).V7.3M.3,.5P1h8.0D. Telefax: (651) 733-1773 TAenlderpehwonMe.: Se(a6c5a1t), 5P7h5.-D3.161 Telefax: (651) 733-1773 0007986 418-012PAGE C2 Protocol 1P8a.g0e1?2 REGULATORYCITATIONS: U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detectionoftoxicittoy reproduction for medicinal products. Federal Register, September 22, 1994, Vol. 59, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministryof Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. European Economic Community (1988). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recom- men the dation on European Ccoommmpulniiatniceesw:itLhepgriislnactiipolne.sof32g(oNoo.d lLab3o1r5a;to2r8y pOrcatcotbiecre).: Official 1-17. Journal of REGULATORYCOMPLIANCE Trheigsulsattiuodnyswcililtebdeacboonved.ucted in compliance with the Good Laboratory Practice (GLP) All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. The Quality Assurance Unit (QAU) will audit the protocol, the raw data and the report, and will inspect critical phasesof the study in accordance with the StandardOperating Procedures of Argus Research Laboratories, Inc. The final report will include a statement signed by the Study Director that the report accurately reflects the raw data obtained during the performance of the study and that all applicable GLP regulations were followed in the conductof the study. Should significant deviations from GLP regulations occur, each will be described in detail, together with how the deviation might affect the quality or integrityof thestudy. `SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT1to the protocol. 000797 418-012,PAGE C-3 Protocol 41P6a.g0e132 TA ESTRTIANCDVEL HICE LE: Identification: Test Article: Name: Physical Description: Lot/Batch Number: Specific Gravity: Purity: Expiration Date: PFOS. Light-colored powder. 217. ~086. 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 identification of Tween 80 to be documented in the raw data. Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the vehicle that would interfere with the resultsofthis study. Therefore, no analyses other than those mentioned in this protocol will be conducted. Safety Precautions: fGolromvuelsa,timoanskpr,epaaprpartoiporniaatnedeaydemipnriosttercattiioonna.nTdhae uMnaitfeorrima/llaSbafceotayt DaarteatoShbeeewto(rMndSuDrSi)ngis attached to the protocol (see ATTACHMENT 2). Storage: Bulk Test Article: Vehicle Components: Prepared Vehicle: Prepared Formulations: Room temperature. Room temperature. Room temperature. Room temperature (samples to be frozen). All test article shipments to the Testing Facility should be addressed to the attention of Julian Gulbinski, Manager of Formulations, at the previously cited address and telephone number. Shipments should include information concerning storage conditions andshipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment. 000798 418-012:PAGE C4 Protocol 41P8a.g0e1s2 FORMULATION: EreqoufPerepnaractioyn: Formulations (suspensions) will be prepared daily at the Testing Facity. Detailed preparation procedures are attached to this protocol (ATTACHMENT 3) AdjustfomrPeurnitty: `The test article will be considered 100% pure for the purposeof dosage calculations. TeFstia ng ciRelseri veSt amply es: The Sponsor the course of will this reserve study. a sample (1 The Testing g)of each Facility will lotofthe reserve bulk test a sample article (5 mL) used during of each lot of tuhnedevrehtihceleprceovmipoousnleynctisteudsceodnddiutriionnsg.the courseofthis study. Samples will be stored IALYS tShaemcpoluersseadodfittihoenasltutdoyt.hose described below may be takenif deemed necessary during BulkTestArticleSampling: INnofoarnmaaltyisonesonoftthheesbtuabliklitteystofartthieclbeuwliklltbesetcaortnidcluectisedondufrilienwgitthhethceouSrpsoensoofr.this study. AnalysesofPreparedFormulations: HHoowmeovgeern,eirteycoarndds wstialbibleitymaoifnptraeipnaerdetdofdoormcuulmaetinotnshoiswotnhefitleeswtitahrttihcleeSfpoornmsuolra.tions were prepared ConceofnTesttArrticale Ftormiulaotionns: sCtoundcye.ntDruaptliiocnaotfe tshaempplreepsa(r2edmLforemauclha)tiwiolnlsbweilltabkeenvefrrifoimedthdeurfiirnsgt athnedcloaustrsperoefparthaitsion roenmtahienidnagysparmepplaeresdw.illObneerseatmaipnleed oaftethaechTessettiwnigllFbaceilsthyipapsedbafcokruapnaslaysmipsl;est.he Backup TseasmtpilnegsFwaiclillbtye ustpoornedreuqnudeesrtotfhethpereSvpioonussolry.cited conditions and discarded at the 000799 418-012:PAGE C-5 Protocol 41P6a.g0e1s2 Shipping Instructions: `Samples to be analyzed will be shipped (frozen on dry ice) to: Kris J. Hansen, Ph.D. 933M5EBnuvsihroAnvmeenntuael Technology and Safety Services Building 2-3-09 TSte.lePpahuoln,eM:inn(e6s1o2t)a77585-163031-83331 Telefax: (612) 7786176 The recipient will be notified in advanceof sample shipment. DISPOSITION: aPrrteicplaerweidlfboermrueltautrinoends twoiltlhbeeSdtiusdcyarMdoenditaotrtahtetTheestpirnegviFoaucsilliytyc.itAeldlardedmraeisnsinugpobunlk test `completion of all work with the test article TESTSYSTEM: `SpecianedRsea/soSnftorrSelaecitionn: `bTehceauNsee:w Z1e)alitainsdonWehintoen-[rHordae:n(tNZmWa)mSmPaFl]iraanbbsiptewcaiesssaeclcecetpetdedasantdhewiTdeesltySuyssetdem ttohxriocuigtyhhoeurtattohgeeniincdiutsyt)r:y2f)orthniosnsctlrianiincaolf srtabubdiiteshaofsdbeeveenlodpemmeonntsatlrtaotxeidcittyo (beembsreynos-ifteitvaelto adenvde4l)optmheentteaslt toxins; article is3)phhiasrtmoariccoallogdiactaalalyndacetxipveeriinetnhcee sepxeisctieast tahnedTsetsratiinn.g Faciity!": Number and Sex Population evaluated: Population selected for toxicokinetic evaluation: Body Weight and Age: 110 timed-pregnant female rabbits (22 per dosage group). l1e9veslasteplliutse tfhermeaelaetrtahbebiottshe(frivdeoastetlheevellso)w. and high dose `rTahbebiitnsdwiivlildubael abpopdryoxwiemiagthetlsy offivtehteofseemavleen rmaobnbitthsswiolfl argaengaet ftrhoemti2m.e5 kg of sttoud5.y5 kg; the dasosciugmnemenntte.d iAnctthuealrbaowddyatwae,igahntdstrheecwoeridgehdtartarnegceeiwpitllabned at study included assignment will be in the final report GC0800 418-012.PAGE C6 Protocol 41P6a-g0e1s2 `Source: Covance Research Products, Inc. D`eSnwvaemrp,brPiedngnesyRlovaadn,iaBo1x77521070 PTrhoedurcatbsb,itsInwci.l,lDbeenvsehri,ppPeednnisnyflevarneida,catrottohnesTbeysttirnugckFafcrioltmy.Covance Research Identification: TRaabgbiCtos,aIrnec.p,eNrom.anMeSntPlTy 2id0e1n0t3i)fi.edFuesmiangleMoranbebiltssaerlef-gpiiverecninugnieqauretpagesrm(aGneeynBtand and giedesnttaitfiiocantiboondnyuwmebigehrtss.when assigned to the study on the basis of day 0ofpresumed ANIMALHUSBANDRY: Al cage sizes Animals. are in compliance with the Guide for the Care and Use of Laboratory Housing: nTehsetirnagbbmiattsewriilallbsewiilnldbiveidsuuaplpllyiheodubseecdauinseunitthse offemsiaxlteoreaibgbhittsstwialilnlbeesssasctreieflicceadgebse.forNeo parturition is expected. RT ooe mAimr,peraatndu Humr idie ty: f`Trheeshanaiirmtahlatrohoasm biseeinndeppaesnsdeednttlhyrosuugphpl9ie9d.9w7it%hHatEPleAasftittteerns.chRaonogemstpeemrpehroautruorfe1wi0ll0%be wmiallinatlasionbeed maton6i1toFre(d16coCn)sttaont7l2yaFn(d2m2aiCn)taanidnemdoantit3o0r%edtoco7n0st%a.ntly. Room humidity Light mAaninatuationmeadt.icaElalychcodnatrrkolpleerdi1od2-whilolurbelgiignhta:t121-9h0o0urhdoaurrksflEuSoTr.escent ight cycle will be: Diet: bApeparvoaxiilmaabtleeltyo e15a0chgroafbbCietrteiaficehd dRaaybbuinttiChthoewfirs#t 5d3a2y2of(PdMoIsaNguetr,itaitonwhIinctehrtnaitmieonal) will Taphperocexritmiafiteedlyfe1e8d0wigllobfethaevasialambelecefrrtoimfieidndfieveidduwailllstbaeinolfefsesresdteteole"aJc-htyrpaebbfieteedaercsh day. attached to each cage. 000801 418-012:PAGE C-7 Protocol 41P8a.g0e1?2 Water: 2Wnataeurtowimlaltbiec wavaatielraibnlge aacdcelisbsitsuymsftreomm. iAnldlivwiadtuealr wbioltltlbees fartotmacaheldoctaol tshoeurccaegeasndorpfarsosmed pthrroocuegshseadrweavteerrseasosamobsacitsermioesmtbatr;apnreocbeesfosreeduwsaet.erCihsleorxipneectwieldl btoe caodndtaeidntnoothme.ore tbhacatner1i.a2lpcopnmtacmhilnoaritnieonatatnhdettwiimceeofanannuaallylsyisf.orWpaostseirblies acnhaelmyizceadl mcoonntatmhinfaotripoonssible Contaminants: 1N0eibteheprretsheenStpionntshoercnerotriftiheed dSiteutdoyrDiinrtehcteodrriisnkaiwnagwreatoefraantylpeovteelnstitahlatcownotualmdininatnetrsferliekely wpietrhftohremerdesbuylttsoheffteheisdsstuupdpy.lieTrhoerretfhoorsee,mneontainoanleydseisn otthhserprtohtaoncotlhowislel broeutcionnedluycted. MAANT DRAI NDOMN IZATG ION s`Tahmeefesmoaulreceraabnbidtsstwriailnl,bebenfaotruerasllhyibprmedetnatottthheeSuTpepsltiienrg, Fbayciibtrye.edTehremadlaey rmaabtbiintsg oofcctuhers cwiolnlsbeecudteisviegndaatyesdadnadys0hiopfpperdetsoutmheedTgeessttiantigonF.aciTlhyeafrtaebrbtihtsewillalstbemamtaitngeddaoyn tfoivaerrive Sounppdlaiyer1,wi2l.l3f,o4rwaorrd5borfepedriensgumreecdorgedsstaatnidon.dayBe0foofreprsehsiupmmeendtgoefsttahteiroanbbbiotds,ytwhee.ights. tAhecormapbubittesrt-ogednoesraagteedgr(owuepigshtb-aosredderoend)thriasnidnofmoirzmaattiioonn. procedure will be used to assign ADMINISTRATION: RR outee andasfoo rChon ice: dTiheetaorryalro(ustet,omtahcehextaubcet)droosuatgeewacasnsbeleeacctceudrfaotreluyseadbmeicnaiusstee:red1;) ainn dco2m)ptairsisoonneowfitthheth.e possible routes of human exposure. MeatndhFreqouendcy: pFreemsaulmeerdabgbeisttsatwiiloln.beDgoisvaengetshewitlelstbearatidcjluesotnedcedadiallyyfoornbdoadyyswe7itghhrtocughhan2g0esofand given at approximately the same time each day. Ratfoir DoosangeSaelelctieon: LDaobsoaragteosriweesr,eInsce.,lePcrtoetdocoonlt4h1e8-b0a1s2iPs).of a dosage-range study (Argus Research 000802 418-012:PAGE C-8 Protocol 41P8a-g0e1s2 foe Le[oe Tox] | DC osao geLn evelc s. entrVa olumtesai ndIo njecn tions Rat, es: sates| oprgtey 2 . [TowToso| 0 | 5 |ecwornonmosmue|n LoTow[orTow | |sumonsonmmee|n FoTaw[v0 1 00 Ts|sononconmmve|n ToRteaasrscrvwginecsornieenede10a0%pare iFuso ofdosagecaioins. NALY: 3 Viability Al Periods: Atleast twice daily. Clinical tionsandlorGeneralA; Predosage Period: Atleast once. Dosage Period: ``Tawpipcreoxdiamialyt.elyProinorethooduorspaogsetdaodsmaignei.stration and once Postdosage Period: Once daly. aClpipnriocpalrioabtseebrvyatthieonSstumdayyDbireecrteocroardneddormoSrteudfyreMqouneinttolry. than cited above, if deemed BodyWeights: Predosage Period: Day 0 atthe ToefsptrinegsuFamceidityg.estation and on the day of arrival Dosage Period: Daily. Postdosage Period: Daily. Feed ConsumptionValues: Predosage Period: Dosage Period: (Rveaclouredsendotdatalbyulaaftteedr).arrival at the Testing Facility Recorded daily. 000803 418-012PAGE C-9 Protocol 41P8a-g0e1s2 Postdosage Period: Recorded daily. Feed consumption values during the dosing period will be tabulated for the same intervals as body weight evaluations. `Caesarean-Sectioning Observations: Rabbits will be Caesarean-sectioned on day 29 of presumed gestation. The fetuses will be removed from the uterus and placed in individual containers. The rabbits will be `examined for number and distribution of: Corpora Lutea. Implantation Sites. d[aPtlaa]centae that appear abnormal (size, color or shape) will be noted in the raw Live and Dead Fetuses. resorption) (Alive fetus is defined as one that responds to stimuli; a dead fetus is defined as a term fetus that does not respond to stimuli and that is not markedly autolyzed:; * dead fetuses demonstrating marked to extreme autolysis are considered to be late (EaArcloynacnedptLuasteisRdeesfoirnpetdioanssa. late resorptionif it is grossly evident that a`onregaarnloygerneseosriptsiohna.)s occurred:if this is not the case, the conceptus is identified as FeOtbaslervations: Body Weights and Idenification: Tbheeusbeoddytowediegthetrmoifneealcihttefrefteutsalwibloldbyewreeicgohrtdeadv.eraOgnelsy. boFdeytu`sweeisghwitlsl obfelitvaegfgeetdusweisthwill identification noting study number, litter number and uterine distribution. `Gross ExternalAlterations All fetuses will be examined for gross extemal alterations. Late resorptions and dead fetuses also will be examined for gross extemal alterations to the extent possible but asnuaclhysoebss.ervations will not be included in either data summarization or statistical 000804 418-012:PAGE C-10 Protocol 4Pa1g8e01120 SE oftxTiassume inataindoSenx: Aevlafleutautseedsiwnilalllbfeeteuxsaemsibnyeddiisnsteercntailolny.toAdestienrgmleincerossesx.-seCcatviiotnawtieldl boergmaansdewilblebteween the parietal and frontal bones, and the brain will be examined in situ. `Skeletal Examination: A`Slkefleettaulsepsrewiplalrbateioenxsamwiilnlebdefroertsakienleedtailn galltyecreartiinownistahfttehrysmtoalinaidndgewditahsalaizparreisnerrveadtiSve. tRaekperne.sentative photographs of fetal gross, soft tissue and skeletal alterations will be MET OF H SACRO IFID CE: Buseeudthtaonsaasciriafi-cDe rSapbebciitasl(v(imaainnutfraacvteunroeudsbiynjSeccthieorn)inagn-dPlloivuegfheAtnusiemsal(vHieaailntthr)apweilrlitboen.eal injection) NECROPSY: Gevraolsusatlieosnio(ncsorwirlelsbpeonrdeitnaginteidssiunensewuitlrlableburfefteariended1f0r%omforrambabliitns ifnortphoesvseihbilceiefuctounrterol gdirsocuaprdaetdt;hethdeissecraerteiocnoofmmtohen,StsupdoyntDiarneectoour)s. le(sEixocnesptiniorna:bbiPtas.r)ovUanrliaensscyssptecsiwfiilclalbley cited below, all other tissues will be discarded atellite Rabbits Assigned to Toxicokinetic Sample Collection `Osnamdpalyes2w1ilofbperceoslulmecetdedgefsrtoamtitohne (rtahbebidtasyafsoslilgonweidngttohtehelatsotxdiocsokaignee)t,ictoexviacloukaitnieotnic wFiolllboewicnogllaencetsetdhfersoima tohfepeinnfteorbiaorrbviteanla, cbalovoadinstaomspelreusm(sapeppraorxaitmoartteulbye4s maLndpceerntrraibfbuigte)d. Tmahientraesiunletdinfgroszeernum(-(7a0ppCr)ouxnitmilatsehliyp2memnLt)towitllhebeSpiomnmseodiraftoerlaynaflryosziesn. on dry ice The liver and wil be erxectiasiende,d watei-g7h0edC,uantnidl sahsiapmmepnlte twoiltlhbeeStpaoknesnofrrfoomrtahnaelyrsiigsh.t lateral lobe, frozen and Rpoasbsbiibtlsewailsl dbeescCraiebseadreaabno-vseecftoriornaebbditasndasfseitgunseeds twiollthbeemeaxianmistnuedyd.grFoesstluysetso tahnedextent placentae wil be pooled Sponsor for analysis. per liter and retained frozen (-70C) until shipment to the Awifltlerbecosmhpilpepteidon(forfozseanmpolnedrcyolilceec)titoon,KrsiesrJu.m,Halnisveerns,ecPthi.oDn.s,, aftettahleanprdevpiloaucselnytaclitseadmples. 00805 418-012:PAGE C-11 Protocol P4a1g8e01121 address advance for of analysis. Both the sample shipment. recipient and the Study Monitor will be notified in ScheduledSacrifice: gOrnosdsayne2c9roopfspyroefstuhmeetdhogreasctiact,ioanb,dfoemmianlael rabbits will and pelvic vbiescCeareasawirlelabne-speecrtfioornmeedd., and a Uteri of aapbpsaernecnetolyfniomnpplarnetgantainont sdioteess".will be stained with 10% ammonium sulfide to confirm the. RabbitsFoundDeador Meribund: dRealbibvietrsy twihlaltbdeieexoramairenesdacfroirfitcheed cbaeucsaeusoefodfeamtohroirbumnodricboundnidticoonn,diatbioorntioonn tohrepdraemyatthuere foebtsuesrevsatainodnloisr mdaedleiv.erePdrepgunpasnwciyll sbteateuxsaamnidneudtetrointehecoenxtteennttspwoislsibbeler,ecuosridnegd.theAsboarmteed wmietthh1od0s%daemscmroinbiedumforsufleftiudseest.o coUtnefriirmoftahpepaarbesnetnlcyenoofnpimrpelgannatnattidonoessitweilsl.be stained 000806 418-012.PAGE C-12 Protocol P41a8g-e01122 PROPOSEDSTATISTICALMETHODS": `Aavpeprroapgrieastea.ndAdpdeirtcieonnatlagpersocweildlubreescaalncdu/loarteadn.alLyitsteesr vmaalyuebsewiplelrbfeorumseedd, wifhaeprpreopriate. I. Parametric A. Bartlett Test* TyofpTeset" Il. Nonparametric A. Kru(sk7a5l-%Watilelss) Test 1 atps0.05 Not T" 7 atps0.05 Not Significant Nonparametric ras Vrars Dunn's Test Significant at|ps0.05 Dunnett's Test Not Significant B. Fisher's Exact Test (>75% ties) Ill. TPesrtfoorportiDoatna VofartihaenBcienTomeisatlfoDrisHtorimbougtieonneity a. b. PStraotpiosrtitciaolnlydsaitganiafriecanntotprionbcalbuidletdieisn tahries rceaptoergtoerdy.aseither p<0.05 or p<0.01. c. d. TUessetdfoornlhyotmooagneanleyizteydoaftvaarwiiatnhceh.omogeneity of variance. 000807 418-012.PAGE C-13 Protocol P4a1g8e01123 DATAACQUISITION,VERIFICATIONAND STORAGE: DDiarteactwoilrlabnedhoarnadp-praonpdr/ioartecommapuntaegre-mreecnotrdpeedr.soRneneclorwditshiwinll21bedareyvsiaefwteedr gbeynetrhaetiSotnu.dyAll boreigbionaulnrdecaonrddsiwnidlelxebde. stAorceodpyinofthaellarrcahwivdeastoafwtilhlebTeesstuipnpgliFeadcitloittyh.eASllpoornisgoinralupdoatna will yreeqauresatf.terPmraeisleirngveodf ttihsesdureasftwiflilnbalersetpoorrte,daafttetrhewhTiecshtitnigmeFatchileitSypaotnsnoorchwiallrgbeefcoornotnaected to determine the disposition of these materials. REC TOO BEMR AIND TAINS ED: TPerosttoAcrotlicalnedVeAhmieclnedmanedn/tosr Reagent Receipt, Preparation and Use. ARnainmdaolmiAzcaqutiisointiSocnh.edules. MVeatteirnignaHriisatonryE.xamination. Treatment (i prescribed General Comments. byStaff Veterinarian) BCllionoicdalSaOmbspelrevaCtoliloencstiaonnd,/PorroGceensesrianlg Appearance. and Shipment (i required). Body Weights. CFaeeesdarCeoanns-uSmepcttiioonniVnagluaensd. Fetal Observations. OGrrogsasn WNeeicgrhotpssy(iObrseeqruviarteido)ns. `PShtoutdoygMraaipnhtsen(iaf nrecqeui(rredo)o.m and environmental records). Feed and Water Analyses. Packing andor Shipment Lists. KEY PERSONNEL: Executive Director of Director of Research: RAelsaenarMc.h:HoMbielrdmraedn,S. PhCh.rDi.s,tiDaAn,BPTh.D., Fellow, ATS ADsirseocctioartoefDLiarbeocrtoartoorfyROepseeraarticohnsa:ndJSothundyF.DiBraercnteotrt:, Raymond B.S. G. York, Ph.D., DABT Manager Manager of of ASntiumdaylCoOopredriantaitoionns: Valerie A. Sharper, M.S. and Member, Institutional Animal Care and MUsae nCoomamfiRtegtgeuele:atDorreynaCoCm.plLieabnoc,eV:.MK.aD.thleeAn. Moran, M.S. Consultant, Veterinary Pathology: W. Ray Brown, D.V.M., Ph.D., ACVP 0003808 418-012:PAGE C-14 Protocol P4a1g8e01124 EINALREPORT. bAecfoimnpalriezheednfsoillvoewidnrgafctofnisnualltraetpioorntwwiiltlhbteheprSepopnasroerd.onThcoemprelpeotritowniolfitnhcelusdteudtyheand will following: `Summary and Conclusion. EEvxapleuraitmieonntaolf TDeesstigRnesaulntds.Method. ADpatpae,ndPircoetsoc:olFiagnudreAss,sSocuimamtaedryAmaennddImnedinvtisduaalnTdaDbelveisatSiuomnsm,arSitzuidnygDtirheecAtobro'sve GLP QAU CSotmaptelmieanntc.e Statement, Reports of Supporting Data (if appropriate) and INSTITUTIONALANIMALCAREANDUSECOMMITTEESTATEMENT: ITnhsetitpurtoiconeadluArneismdaelsCcrairbeedanidn tUhsisepCroomtomciotltehea.veAblleeprnorceevdiuerweesddbesyctrhiebeTdesitnitnhgisFapcriolttoyc'osl thatinvolve discomfort, dsitsutdryesasnoirmaplasinwitlol tbhee caonnimdaulcst.ed in a manner to avoid or minimize `nTehceesSspiotnysoforr'scosnidguncattiunrge tbheisloswtuddoycaunmdentthse ftahcetftahcatttthhaits iisnfnoortmaatniounnnceocnecsesranriinlgythe. duplicative procedures study were amvaayilbabeleobftoarimneeedtfirnogmtthheesStpaotendsopru.rpNosoeaslotfertnhaetisvteud(yi.n vitro) REFERENCES 1. oChfrtihsteiatne,ratM.oSg.eniHcopboetrenmtainal,oAfMs.usapnednsSimointsh,ofTt.rHi.nFi.tr(o1f9l8u2c)r.enoDnoes(aTgNeF-)range study `administered orally to New Zealand White rabbits. Toxicologist 2(1):40 (#143). 2. nCharlitsrteixaon,neM.(SP.ro(c1e8e8d4i).ngsReofprNoadlutcrteixvoenetoSxiycmiptyosainudmt,erNateowloYgoyrekvaAlcuaatdieomnsyooff Sciences, November 7, 1983), J. Clin. Psychiat. 45(9):7-10. 3. MF.eSu.ss(n1e9r8,2).EL.A dLeigchatdkeepo,fGr.aEb.b,itHfeenrnteltsyya,tRaA:.,StHuodbyerofmahnis,toAriMc.al aconndtrCohlriasntiimaanl,s. Teratology 45(4):349-365. 4. LInasbtoitruatteoorfyLAanbiomarlast.oryNaAtniiomnaall ARecsaoduermcyesPr(e19s9s6,).WasGhuiindgetofnor,tDh.eC.Care and Use of 5. TSetrapalteosl,ogRyE9.(3()1:9A7347)-.38.Detection of visceral alterations in mammalian fetuses. 000809 418-012:PAGE C-15 Protocol P4a1g8e01125 6. tShtaepKlOeHs-,aRl.iEz.arainndreScdhSnelmle,tVh.oLd. f(o1r9f6e4t)a.l bRoenfei.neSmteanitn Tinecrhanpoild.cl3e8a:r6i1n-g63t.echnique in 7. ISmaplleawnstkait,ioE.nss(1t9e6l4l)e.n aFmarUbteemreutshdoedreRaztutem. maArkcrho.skPoatphioslc.hEexnp.NaPchhawremaiksolv,on 247:367. 8. tShneedbeicnoorm,iaGlWdi.strainbudtiCoon.chSrtaant,isWti.cGa.l M(e1t9h67o)d.s,V6atrhiaEdnicteiotne,stlfoowrahSotmaotgeenUneiivteyrsoifty Press, Ames, pp. 240-241. 9. BSoikoamle,trRy,R.W.aHn.dFRroehlefm,aFn.Ja.n(d18C6o9.),. SBaanrtFlreatnctitsecsot,opfph.o3m7o0g-e3n7e1ityofvariances. 10. MSneetdheocdosr,,6tGh.WE.ditainodn,CloocwharaSnt,atWe.UGn.iv(e1r9s6i7t)y.PrAensasl,ysAimsoesf,Vaprpi.an2c5e6.-27S5t.atistical 11. tDruennaettmte,ntCs.wWi.th(1a95c5o)nt.rolA. muJ.ltAimpelre.coSmtapta.rAisssooncp.r5o0c:e1d0u9r6e-1fo1r29c.omparing several 12. WSoHk.al,FRrRe.emaanndanRodhiCfo,.,F.SJ.an(19F6r9a)n.cisKcrou,skpap.l-W3a8l8l-i3s89T.est. Biometry, 13. 6D(u3n)n:,24O1.-J2.52(.1964). Multiple comparisons using rank sums. Technometrics 14. Siegel, S. (1956). McGraw-Hill New YNoornkp,arppa.me9t6r-i10c4.Statistics for the Behavioral Sciences, 000810 PROTOCOLAPEROVAL: FOR THE TESTING FACILITY / George E, Assotiate fe arlove, Ph.D., DABT Director of Research R d G. York, AsSsttudy DitreecDtiorrector of de . DABT Research Dis Cofoidoe Dena C. Lebo, V.M.D. Member, Institutional Animal Care and Use Committee )FOR THE SPONSOR sea To Marvin T. Case, D.V.M., Ph.D. Study Monitor 418-012:PAGE C-16 Praca srroaon lpeb-7p Date fideo Date ot e Date 7 Phyge Date 000811 418-012:PAGE C-17 ATTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE 000812 ATTACHMENT 1 418-012:PAGE C-18 ProtocPola4g1e8.1001122 STUDYSCHEMATIC RABBIT DEVELOPMENTAL TOXICITY STUDY* DSotasratgoef TimeRdaPbrbeigsnant PDreas7yumoefd Gestion Caesarean: DEonsdaogr`eSectioning PDraeys2u0m0e1d Gestation PDraeys2u0meodf Gestation aW=E F-oDroasdadgiteioPnearlioddetails protocol. see "Tests, Analyses and Measurements" sectionof the b= Fetal evaluations (external, softtissue and skeletal). 000813 ATTACHMENT 1 418-012:PAGE C-19 ProtocFolas4e18z.a0r1s2 21AUG 98 23 AUG 98- 09 SEP 98 06 SEP 98 - 10 SEP 98 14 SEP 98 - 18 SEP 98 17 DEC 98 SCHEDULE* Animals Arrive -Acclimation Begins. Dosage Period (Days 7 through 20 of presumed gestation). Toxicokinetic Sample Collection (Day 21 of presumed gestation). Caesarean-Sectioning Period (Day 29 of presumed gestation). Draft Final Report. a. The study initiation date is the date the Study Director signs the protocol. } 000814 418-012.PAGE C-20 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 00081% MATERIAL SAFETY DATA SHEET 418-012,PAGE C-21 am 3M Center sSts.iesP-a1u0l,00Minnesota 1-800-364-3577 or (812) 737-6501 (24 hours) ACLoLpyrriigghhtt,s r19e9s8e,rveMdi.nnesCootpayiMnignianngd/oarnddoMwannluofaadcitnugrinogf tChoimspany. iisnfoarlmlaotwiedonprfoovridtehde tphuartp:ose of properly utilizing SM products 1) pthreiorinafgorremeamteinotn iiss ocbotpaiiednedinfrfoumllSHw,ithandno changes unless 2) dneiistthreirbuttehde wciotphy tnhoer tihneteonrtiiogninoafl eiasrnriensgolda porrofoitthetrhweirseeon. TDRIAVDISEIORNA:NE:3M CHEMICALS 10FCN-U9M5BEFRL/UUO.RPA.DC.:Brand Fluorocheaical Surfactant 9988.-00221017..00818063..57 0000.-5511113355.-0099306542-.17 9988..00221017--30911064.-15 ISSZUFE.Do:o0J2an1u0aary 29, 1L 998 L DSOUCPUEMRESNETD:ES:10N-o3v7e9m6b-e9r 05, 1967 0000.-5511113355--0029301515.-28 1. INGREDIENT CAS. NO. PERCENT PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SSUULLFFOONNAATTEE........... PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SSUULLFFOONNAATTEE............ POTASSIUM PERFLUOROALKYL SULFONATE...... 3278957-139.-939.6382."e86 6200247200--5459.-53 32 T"e7 3872-25.1 1 Ts 2. PHYSICAL DATA vBaOpIoLRINGPPROINET: .SL.L.S.L..U.1..R1.1.E.1.1.S.1. WNIaA vEaVpAoPRORApeTnIsOtNTRYA:TE: 111.1101 1111110 wN/aA SSPOELCUIBFIILCITYGRAIVNITWY:.A..T.0...E.1R11.1:11]] csal.ign0t.6 Water=1 PERCENT VOLATILE:.............. 0(%Bulk) HVEICSTCIONSGITPYo:.i.n.:..L..1.1o1ie. NI(D0.1% Win Aqueous) APPLEiAghRtANCcEoloArNeDd,ODOfRr:ee flowing pouder. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 000816 MJSaDnSu:aryFC-299,5 F1L9U98ORAD Brand Fluorochenical Surfactant 418-012:PAGE C-22 PAGE 2 3. FIRE AND EXPLOSION HAZARD DATA FFLLAASRHMPBOLIENTL:I.N.T.T.S..L..E..E...1..1.I..T. FLAMMABLE LINITS - UEL:.. 0.0. WNoAne N/A AUTOIGNITION TEMPERATURE: ...... N/A EXWTaItNeGrU,ISHCIaNrbGonMEDdIiAo:xide, Dry chemical, Foam SPWEeCaIrALfuFlIlREprFoItGeHcTtIiNvGePRcOlCoEtDhiUnRgE,S: including helmet, self-contained, apnodsitpiavntes,prebsasnudsrearoorunpdresasrumsr,e wdaeimsatndanbdrealteghsi,ngfaAcppearmaatsuks,, anbdunker coat protective covering for exposed areas of the head. UNSUeSeUALHazFaIRrEdouAsNDDeEcXoPLmOpSoIsOiNtioHnAZAsReDcSt:ion for products of combustion. 4. REACTIVITY DATA STABILITY: Stable INNCoOtMPAaTpIpBliIcLaIbTlYe.- MATERIALS/CONDITIONS TO AVOID: HAZARDOUS POLYMERIZATION: Hazardous polymerization Will not ocour. HAZCAarRbDoOnUSMoDnEoCxOiMdPeOSIaTndIONCarPbRoODnUCDTiSo:xide, Oxides of Sulfur, Hydrogen Fluoride, Toxic Vapors, Gases or Particulates. "5. ENVIROMENTAL INFORMATION Tr SPOILbLserRvESePOpNrSeEc:autions from other sections. Vacuum, use Wet sweeping cboempaonunidgniortioantesrourtcoe.avoiCdleadnustuipngr.esCiAdUuTeIOwNi!thAwavtaecru.um cPllaecaenerincoanuld approved metal container. Seal the container. RECDoOMMnoEtNDErDeleDaIsSePOStAoL:Waterways or sewer. Do not use in products or pTr/1o0cesosfesthethaltowecsotuldECSrOesuolrtLCiSnOaqcuonacteinctrcaotnicoenn.tratIinocnisnegrrateeateinr tanhan imantdeursitarli.al CoormbcuosmtmieornciaplrodfuacctisliwtiyllinintchleudpereHsFe.nceDiofspoasaclombustible alternative: Dispose of waste product in a facility permitted to Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 000817 JJSaDnSu:aryFG-299,5 1F9L9U8ORAD Brand Fluorocheaical Surfactant 418-012:PAGE C-23 Pace 3 5. ENVIRONMENTAL INFORMATION (continued) accept chemical waste. enBvoIhoRuOnrWgIiEaNATqA`wLttBaDeAoThAuF:(iLsenhpoLatCiSsD, maFactrhoeoahdiruWsiyn~ncohu(aiga/s1p,haRlaeisnbopwroTsreoluats)(=s3a8lemng/L, SGaoirrinBeorosz)eer=s niBtO.IL] 8-H. ECS0, DAPARLA Magna = 80 ma/L;COD -004 REGVVUooLlcAaTtLOiaRlsYes MOIfENgDFaORn3MiAoETxIeCOnoNpm:tpouSnodlsv:entNsI:A. WIA. oSBiehnfcoerheazraderigdsuopluoasst)ailo.ns Uvasr.y, EPcAonNsauzltardaopopsliHcAaSbTlee NroeRgbuOlrati=onsNonoer a(uNtOThorUi.St:ies TThoihs, prEoindEuccst,coCosopLl,issAICWSi,thWItThSe acnhdemiKcoarela.registration requirements of ErTcIaRnE WAAzZAaRmD:LANeS. S:PRESSURE: No REACTIVITY: Mo AGUTE: Yes CHRONIC: Yes "6. SUGGESTED FIRST AID evieTnacusetodeiesa.rte:looytf1Aumsmhedieaytees mweidtihcallargSerteanmtoiuonnt:s of water for at least 15 skciTnoammtceaodnnitisantceettl:yadflSulsohthisnkgi.n wIifthArFliarrgaetioanmoupnetrseieorfs,watGeari.l aRemporvyeeician. Mash ety tren IASTLfiAGTrSeiIgrOneNsy:uspytmopntsomssentoicmcuuer,, creamlolvae pperryseoinsiatno fresh air. It IFDrsuunAkLLOwHeED:Glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION : evevopirdoreecyteiocno:ntact. Wear vented goggles. Aobreviations: NID - Not Determined WIA . Not Applicasie Gh - Approximately . 000818 MSDS: FC-95 FLUORAD Brand Fluorochesical Surfactant January 29, 1998 418-012:PAGE C-24 PAGE 4 7. PRECAUTIONARY INFORMATION (continued) SKAIvNoiPdROTsEkCiTnIOcNo:ntact. Wear appropriate gloves when handling this mraetceormimaeln.ded:A pabiutrylof rugblboevre.s maUdsee ofnreomorthemofroelloofwitnhge fmoaltleorwiianl(gs) are cpoevresroinnagl, prcoovteercatlilosn. itePmrsoteasctinveecesgsaarrmyenttso p(roetvheenrtthsakinn gcloonvetsa)ct:shouhledad pbeolymeatdheyloefnee/iptohleyrvionfyltihdeenfeollcohwlionrgidemat(eSrairaalnse:x). RECUsOeMMEwNiDtEhDaVpEpNrToIpLrAiTaItOeN:local exhaust ventilation. Use in a wellevemnitsisliaotnesdbearleoaw. rePcroomvmiednededsuefxfpiocsiuernet lviemnittisl.atioInf etoxhmaauisnttavienntilation is not adequate, use appropriate respiratory protection. RES`APvIoRiAdTObRrYeatPhRiOnTEgCToIfONd:ust. Select one of the following NIOSH approved respirators based on airborne concentration of contaminants and in hacaclofr-dmaanscke swuiptphlieOdSHaAirregruelsaptiiroantso:r, fhualllf--fmaacsek dduusstt aanndd mmiisstt rreessppiirraattoorr,, full-face supplied air respirator. PREVENTION OF Do not eat, AdCrCiInDkENoTrALsmoIkNeGEWSThIeOnN:using this product. Wash exposed bareefaosretheaotrionugg.hly With soap and water. Wash hands after handling and RECKOeeMpMENcDonEtDaiSnTeOrRAGdEr:y. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: Nonflammable. OTHNEoRsmPoRkEiCnAgU:TIOSNmAoRkYingINWFhOiRlMeATIuOsNi:ng this product can result in ocfontthaeminhaatziaorndouosf dtheecotmopboascictoionandp/roorducstmsokemeanntdionleedadintosethcetiofnor4matofion this MSDS. HIS HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.) EXPOSURE LIMITS INGREDIENT VALUE UNIT TYPE AUTH SKIN POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 THA aM Y PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SSUULLFFOONNAATTEE...... 00..11 MMGG//MM33 TTHHAA aM Y POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 THA 3M Y Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 000819 418-012:PAGE C-25 JMSaDnSu:aryFC-299,5 1F9L9U8ORAD Brand Fluorochemical Surfactant PAGE 5 EXPOSURE LIMITS (continued) INGREDIENT VALUE UNIT TYPE AUTH SKIN POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 TM wm t=heSKIpNoteNOnTtAiTaIlONc:ontrLiibsuttedionsubtsotatnhceesoviernadlilcateexdposwuirteh b'yY' tuhendecrutSaKnIeNousrefreoruteto biynclduidriencgt cmounctoaucst mweimtbhranteheansdubseytea,ncee.itheVrehbiycleasirbcoarnnealtoer,r more skin apabrstoircputliaornl.y, SZO'U3MR:CE OF3MEXRPeOcSoUmRmEenLdIeMdITExDApToAs:ure Guidelines 8. HEALTH HAZARD DATA EYEMilCdONTEAyCeT:Irritation: signs/sysptoss can include redness, swelling, pain, and tearing. SKIN CONTACT: Mild Skin Irritation (after prolonged or repeated contact): signs/symptoms can include redness, swelling, and itching. eMaxytenbdeedabtsiomreb.ed through the skin and persist in the body for an INMHaAyLATbIeONh:armful if inhaled. May be absorbed by inhalation and persist in the body for an extended Time. Single overexposure, above recommended guidelines, may cause: sIorrreinteastsionof (tuhpepernosreespainrdattohrryo)a:t, sicgonusg/hsiyngmptaonmdssnceaenziinngc.lude IF_ISnHgAeLsLtOiWoEnD:is not a likely route of exposure to this product. Illness may result from a single swallowing of a moderate quantity of this material. May be harmful if swallowed. MUTMAuGtEaNgIeCnIiTcYi:ty assays indicate the product is not mutagenic. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 000820 MSDS: FC-95 FLUORAD Brand Fluorochemical Surfactant January 29, 1998 418-012:PAGE C-26 PAGE 6 8. HEALTH HAZARD DATA (continued) REPNRoOtDUtCeTrIaVtEo/gDeEnViEcLOiPnHEtNhTeALratTOXaItNSo:ral doses below maternally toxic Levels. OTTHhEiRsHEprAoLdTuHctHAZisARnDotINkFnOoRwMnATItOoN:contain any substances regulated under California Proposition 65. A Product Toxicity Susmary Sheet is available. SECTION CHANGE DATES HEADING SECTION CHANGED SINCE November 05, 1997 ISSUE Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately bTehecoirnrfeocrtmataisonofinthethdiasteMatiesrsuieadl. Saf3eMtyMAKDEaStaNOShWeAetRRA(NMTSIDESS), isEXbPeRlEiSSeEvDedORto MIEMRPCLHIAEDN,TABIINLCILTUYDINOGR,FIBTUNTESNSOTFOLRIMAITEPDARTTIO,CULAANRY PIMUPRLOISEED OWRARRCAOUNRTSYEOFOF WPhEeRtFhOeRrMANthCeE 3OHR pUSrAoGdEuctOFisTRfADiEt. forUsearpaisrtirceuslpaornsipbulreposfeoranddetesrumiitnaibnlge for cuasner'asffemcetthotdheofuseuseandorapappplliiccaattiioonn.of aGi3vHenprtohdeucvta,riestoymeofoffwahcitocrhsartehat tuhneiquuesleyr ewviatlhuiantethetheus3eMr'sprokdnuocwtledtgoedeatndermcionnetrowlh,ethietrisitesissenftitialforthaat particular purpose and suitable for user's method of use or application. D3Mueprtoovitdheesreimnoftoermaptoisosnibiinliteylectthartonieclefcotrromniacs tarasnesrfveircematyo hitasvecursetsoumletresd. rinepreersroernst,atioomnisssiasonstooritsaltceormaptlieotnesnesisn otrhisacciunrfaocrym.atioInn, ad3d4itmiaokne,s no information obtained from a database may not be as current as the information in the MSDS available directly from 3M. 000821 418-012:PAGE C-27 ATTACHMENT 3 TEST ARTICLE PREPARATION PROCEDURE 000822 418-012:PAGE C-28 ATTACHMENT 3 Version: 418-P0r1o2to(c0o9lA4U1G809182) TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE Page 1013 Test Article: PFOS Vehicle: 0.5% Tween 80 in R.O. Deionized Water A. Purpose: TofhdeopsuargpeosseuosfptehnissiopnrsocoefdPuFreOiSs taondprtohveidveehaicmleetfhoordorfaolratdhmeinpirsetpraartaitoinonto rabbits on Argus Study 418-012. B. General Information: 1. Aslplecsiufsypethnesipornotcoocnotlaniunemrbserwi,lltebset laratbieclleedidaenntdifcioclaotriocno,dAerd.gusEbaacthchlabel will naunmdbsetro,racgoencceonntdriattiioonsn., dosage level, preparation date, expiration date 2a. Suspensions will be prepared: X_ Daily __ Weeky 2b. Vehicle will be prepared: Day X_ Weey _For__daysofuse _For__daysofuse 3. Suspensions will be prepared at a final dosage volume of 5 mL/kg. 4 safety: X_ XC GDulsotv-eMsi,stlaRbecsopaitr,agtoorggles or safety glasses and faceshield _ Half-Face Respirator TFyuvlekkFaScueitR/eAsppriorantor/Positive Pressure Hood 5. --DosageYessuspensions adjusted X_ No for Free base and% (Calculations based Puriy. on 100%) T_ FreeBase __ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol. 000823 418-012:PAGE C29 ATTACHMENT 3 Version: 418.0P1r2ot.o(c0o9lA4U18G.09182 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE Page 2013 NOTE: tTheestloarwtidcolseawgiell.beOnprceepatrheedfianaslavoselruimalesdialruteioanchfireovmedt,hesthiirgbhadrsosaargeetotobe `adamdidneidsttroattihoen.containers; mixing should occur during sampling and/or C. Preparation of Vehicle 1. AIadbdeltehdecroenqtuaiirneerd.amHoeuantttohfe Rw.aOt.erdteoio5n0izCe,dw+a5teCr,taodadntahpeprreoqpuriiraetdely amount of Tween CALCULATIONS) 80 and mix until uniform (See TEST ARTICLE D. Test Article Suspension Preparation 1. Taomporuenptaorfetteshtea0r.ti7c5lem(gS/emeL,TEGSroTuApRVTIsCusLpEenCsAiLonC,ULadAdTIthOeNSre)quiinrteodan `vaephpircolperiaantdelyhesaitzetdh,e lmaibxetluerdectoont8a0ineCr.+5QSC ad for to the required approximately amount with 30 minutes or until the TAJS dissolves. 2. Osunrceethtehreeteisstaavritsicilbelehvaosrtdeixs,sotlhivsedw;illspaicnhiwheivleetthheedseussipreednseimounlsciooonl.s.Th(iBse may be prepared the day before use.) 3. `Taomopurnetpaorfestthoeck0.s5usmpge/nmsLi,onG(rGoruopuIpVVs)u(sSpeenesiToEnS,TrAeRmoTvIeCLthEe required CALCULATIONS). QS ad with the vehicle and mix. 4. aTomopurnetpaorfestthoeck0.s2usmpge/nmsLi,onGr(oGruopulplIsVu)s(pSeenesiTonE,STreAmRoTvIeCtLhEe required CALCULATIONS), QS ad with the vehicie and mix. 5. Tamoopurnetpaorfestthoeck0s.0u2spmegn/smiLo,n G(GrroouuppIlIs)us(pSeenesTioEnS,TrAemRoTvIeCLthEe required CALCULATIONS), QS ad with the vehicle and mix. 000824 418-012PAGE C-30 ATTACHMENT 3 Version 418.P0r1o2t.oc0o5lA4U18G.06182 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE Page 3013 6. vTeohipcrleeptaoreanthaepp0rompgr/imaLte,lyGrsoizuepd,| lsaubsepleendsicoonn,taaidnedrr(eSqeuierTedESamToAuRnTtIoCfLE CALCULATIONS) and mix. Wiiten by: _ "Mak (. Approved by/z Date: _=5-9F Clarification: "X_No __"ds (See attached clarification form) Initals/Date : Ly 29 SEV GF 000825 2PRIMEDICA _----m 418-012:PAGE C-31 ER SHoBrsy heam,bPlA 1i90r44 PROTOCOL 418-012 ORAL (STOMACH TUOBFE)PFDOESVEILNORPAMBEBINTTSAL TOXICITY STUDY SPONSOR'S STUDY NUMBER: 6295.10 Amendment 1 - 11 December 1998 emememrrreeeeeeeeeeeeeseessmmmeeeeeeeeeeemeeeeeees 1 Sponsor (page 1ofthe protocol): The Sponsor is 3M Corporate Toxicology,ratherthan 3M Toxicology Services. Reason for Change: This change was made at the request of theSponsor. 2. Species/Strain and Reason for Selection (page 5of the protocol): sotrsaitn.est article is biologically active, rather than pharmacologically active in this Reafos rChoangne: This change was made at the requestof the Sponsor. 3. RR outee andasfoo rChon ice (page 7of the protocol): oTnhee porraolpo(gsaevdagfoer)crlionuitcealisusae.possible route of human exposure, rather than the 000826 Rea forsCho angne: This change was made at the requestofthe Sponsor. 418-012PAGE C32 ProtAocmoeln4d1m8e.0n1t12 Page2 iil (fom JAD ones n i. Hoberman, Ph.D., DABT Date `Reyfabnd G. York, fo DABT Date Director of Research aAsnsdocSitautdey DDiirreeccttoorr &f Rsearch Ubee-s 7 Dena Chairperson, , M.D. Institutional Animal Date Care and Use Committee Manes: Tlaas Manin T. Case, Study Monitor D.V.M., Ph.D. 7d Date 000827 APPENDIX D PILOT REPORT 000828 418-012.PAGE D-1 FINAL PILOT REPORT `Study Title Oral (Stomach Tube) Dosage-Range Developmental Toxicity Study of PFOS. in Rabbits SPONSOR'S STUDY NUMBER: T-6295.10 Author Raymond G. York Ph.D., DABT (Study Director) Stud leted 6 January 1999 (Final Pilot Report) Performing Laboratory Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 LaboratoryProjectID Argus Research Laboratories, Inc., Protocol Number: 418-012P 000829 418-012:PAGE D-2 PROTOCOL 418-012P: 418.012PPAGE 2 ODERVAELL(OSPTMOEMNATCAHLTTUOBXEI)CIDTOYSASGTEU-DRYAONFGPEFOS IN RABBITS SPONSOR'S STUDY NUMBER: T-6295.10 SUBJECT ABSTRACT IL Purpose TAOFB CONL TENE TS I. Methods nn. Results Iv. Conclusion Figure 1. Table 1. Table 2. Table 3. Maternal Body Weights Clinical Observations - Summary Necropsy Observations - Summary Uterine Contents for the Individual Rabbits that were Found Dead or Aborted Table 4. Table 5. Table 6. Maternal Body Weights - Summary Maternal Body Weight Changes - Summary SMautmemranarlyAbsolute Feed Consumption Values (g/day) - Table 7. VMaalteurensa(lgRheklga/tdiavye) -FeSeudmmCaonrsyumption PAGE 4 6 6 7 12 13 14 16 20 22 26 2 30 000830 418-012:PAGE D-3 SUBJECT 418-012PPAGE 3 PAGE Table 8. Caesarean-Sectioning Observations - Summary 32 Table 9. Litter Observations `Summary (Caesarean-Delivered Fetuses) - 34 Table 10. Fetal Gross External Alterations - Summary 36 Table 11. Clinical Observations - Individual Data 38 Table 12. Necropsy Observations - Individual Data 42 Table 13. Maternal Body Weights - Individual Data 45 Table 14. Maternal Feed Consumption Values - Individual Data 52 Table 15. Caesarean-Sectioning Observations - Individual Data 59 Table 16. Litter Observations Individual Data (Caesarean-Delivered Fetuses) - 61 Table 17. Fetal Sex, Vital Status and Body Weight - Individual Data 63 ATTACHMENT 1 - PROTOCOL 66 000831 418-012PAGE D4 418.012P PAGE 4 TITLE: ORAL (STOMACH TUBE) DOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY OF PFOS IN RABBITS APRRGOUTSOCROELSENUAMRBCEHR:LAB41O8R-A0T1O2RPIES, INC. SPONSOR'S STUDY NUMBER: T-6295.10 ABSTRACT Trhainrdtyo-mflivyeapsrseisgnuemdedtopsreevgenanndtoNseagweZgeraoluapnsd, Wfhivietpee[rHgrrao:u(pN.ZWS)uSsPpFe]nsriaobbnistsofwere PFOS were administered orally once daily on days 7 through 20 of presumed gestation (DGs7to 20) at dosages of 0 (Vehicle), 0.1, 1, 2.5, 5, 10 and 20 mg/kg/day. The vehicle processed deionized water. wTahse0d.o5s%agTeweveonlum8e0wian sre1v0ermsLe/okgs,moasdijsumsetemdbdraailnye on the basis of the individual body weights. Checks for viability were made twice daily. Clinical observations wererecorded twice daily during the dosage period (once prior to dosage administration and once approximately one hour after dosage administration) and once daily during the postdosage period. Body weights and feed `consumption values were recorded daily during the dosage and postdosage periods. All surviving rabbits were sacrificed on DG 29 and examined for the number and dniesctrriobpustyioonfotfhceotrhpoorraacilcu,teaab,diommpilnaanltaatniodnpseiltveiscavnidscuetrearwinaescopnetrefnotrsm.ed.AgrFoestusses were weighed and examined for gross external alterations and sex. dgTrowosouapgdeoaebgsorroitnueptdhwaeen5rdemwgfe/orkuegn/dsdadcareyiafdidcoeasdn.adgeAodngdreitoiduoopneaalilnnyd,tfhfoeouursraidnmoetehsde oi1ns0athgmege/2gk0rgo/mudgpa/ykagbd/oordstaaeygde and was sacrificed. oocbcsuerrrveadtiionnsth(es2c.a5n,t Dosage-dependant increases in adverse 5o,r 1n0oafnecdes2,0umngg/rkogo/dmaeyddcoosatagaendgreouxpcse.ss clinical salivation) Rabbits in the 10 and 20 mg/kg/day dosage groups had body weight losses and severely reduced absolute and relative feed consumption values for all recorded 000832 418-012:PAGE D-5 418-012PPAGE 5 intervals after these groups initiation of dosage; there were no after DGs 26 and 19, respectively. surviving pregnant rabbits The 2.5 and 5 mg/kg/day in ednotsiraegedogsraoguepsanhdadgedsetcarteiaonsepderbiooddsy. wAebisgohltugtaeinasndorrbeloadtiyvewefiegehdtcloonsssuemspftoirotnhe vdaolsuaegsfeorgrtohuepse.ntFireeeddocsoangseumppertiioodnwvearleuersewduecreedriendtuhceed2.i5n atnhed 25.5mga/nkdg/day 5 mg/kg/day dosing. dosage groups for the entire gestation period after the initiation of Freetsaolrpbtoiodnyswaenidghtthsewpeerrecesnetvaegreeloyfrceodnucceepdtuasnedstpheernluitmtebrewreroef eianrclryeaasneddliantethe 5 mg/kg/day dosage group, however, only two of dosage group survived to Caesarean-sectioning. the four pregnant does in Fetal body weights were this reduced controls in the 2.5 mg/kg/day but were at the lower dosage group as endof the range compared to the concurrent observed historically at the TuneasftfiencgtFeadciblytya.dmAilnliostthreatriCoaneosfarteheant-esstecatritiocnlienagt odosliatgerespaarsamheitgehrasswere 5 mg/kg/day. iOnnciesofrest.usTiwn othfeet2u.s5emsgi/nktgh/eda0.y1 dmogs/akgge/dgaryoudposhaagdeagcrloefutpshnaodutfeatnald garbossesnt malformations. other fetus had One fetus abdominal had a short distention. snout, displaced and small nares and the BPaFsOeSd woenrtehreseecodamtmae,nddoesdagfeorstohfe0de(Vveehliocplem)e,nt0.a1l,t1o.x0i,ci2t.y5staunddy3i.n7r5abmbgi/tskg(/Adragyus Protocol 418-012) 000833 418-012:PAGE D6 I. Purpose: 418-012PPAGE tTohbeepuusrpeodsien otfhethdisevsetluodpymweantsatlotporxiocviitdye(ienmfborrymo-afteitoanlfotroxtihceitysealnedctitoernaotfogdeonisca.ges Wpohtietnetia[lH)rast:u(dNyZWo)fSPPFFO]Spraedsmiunmiesdteprreedgnoraanltlyfveimaalsetormababciths.tube to New Zealand Il. Methods 2T0heMtaeyst1a9r9ti8c,lea,nPdFwOaSs(sLtootr2e1d7a)t, raonoomfftwehmipteeraptouwrdee.r,Thweasverheiccleei,ve0d.o5n% Tween80 was stored (lot MO3HOS), a clear at room temperature. viscous fluid The vehicle was received on 1 July 1988, diluent, reverse osmosis and fmreommbaracnonetipnruoocuesssseodurdceeioantitzheed Tweastteirng(RF.aOc.iliDteyiaonndizweadsWamtaeirn)tawiansedaavtairloabolme temperature. Trhainrdtyo-mflivyeapsrseisgnuemdedtopsreevgneanndtoNseagweZgeraoluapnsd White [Hra:(NZW)SPF] [five per group (Groups rabbits were. | through oVinb]d.aySsus7ptehnrsoiuognhs2o0foPfFpOrSeswuemreedagdemsitnaitsitoenr(edDGosral7lyto(2s0t)omaatcdhotsuabgee)soonfce daily 0 (Vehicle), 0.1, mLkg, adjusted 1, 2.5, 5, 10 daily on the and 20 basis of mtgh/ekgi/nddiavyi.duaTlhbeoddoyswaegieghvtoslruemceorwdaesd 10 immediately before intubationof the test article. Checks for twice daily dvuirabiinlgittyhweedroesmaagdeepetrwiiocde daily. (once pCrliionrictaoldoobssaegrveaatdimoinnsiswterraetiroencaonrdded tohnecepoasptpdroosxaigmaetpeelryiood.ne Bhoodury awfetiegrhdtossaangdefaedemidncisotnrsautmiopnt)ioanndvaolnucese wdaeirley during recorded daily during the dosage and postdosage periods. Adlilstsruirbvuitvioinngofracbobriptsorwaelruetesaa,criimfpilcaendtaotnioDnGsi2te9s and and examined for the uterine contents. number and A gross wneecrreowpesiygohfetdheantdhoeraxcaimc,inaebddofmoirngarlosasnedxtpeerlvniacl vailstceerartaiwonassapnedrfsoerxm.ed. Fetuses a aDreetapirloevdiddeesdcriinptthieonasttoafcahlledprporcoetdocuorlesanudseadmeinn dtmheenctosn.ducDtevoifatthiiosnsstfurdoym the are PavraoitloacbolleainndthSetarnadwadradtOaperating Procedures of the Testing Facility 000834 418-012:PAGE D-7 Wl. Results: 418.012PPAGE 7 A and i icaalnd 2; Individual Data Ne - Tables 3. 11 ations and 12) (Summaries -Tables 1 A. Mortality, Abortion: terine Contents of Aborted Rabi (T8w2o61d,o8e2s62(,8285376a4nadn8d258296)5i)n itnheth5e m1g0/mkgg//dkagy/ddaoysdaogseaggreogurpoaunpdafboourrteddoeasnd 2w0ermeg/sakcgr/idfiacyedd.osAdadgietigornaolulpy,wfeoruer fdoouensd(d8e2a66d,a8n2d67o,ne82d6o8ea(n8d26892)7i0n)tihnetshaeme idnotsuabagteiognroacucpidaebnotr.teTdhaensdewdaosessaacrreifdiecesdc.riTbhede bdeelaotwh.ofAldlooeth8e2r6d6owesassudruveivteodatno scheduled sacrifice. malkalday dosage DDoGe2832a5f7teirn 1t4heda5ilmygd/oksga/gdeasy.doTshaegeonglyroaudpvearbsoertceldinaicnadl owbassersvaactriiofincwedasonscant rfeescoersp(tiDoGnss w1e3r1e0a2b2)o.rteNdoagnrdososnelelsaitoensrewseorrpetrioenvewaalsedibnyutnereoc.roTphsiysadnodefliovsetlate body weight period. and feed consumption values were reduced throughout the dosage DDoGe2822a5f9teirn 1t4heda5ilmygd/oksga/gdeasy.doTshaegeonglyroaudpvearbsoertceldinaicnadl owbassersvaactriiofincwedasonscant dfeoceesab(oDrGtesd1s4evaennd d1e6a1d0 f21e)t.useNsoagnrdostswolelsaitoenrsewseorrpetiroenv;eanloedcboyncneepctruopsseys. This remained in utero. reduced after DG This 11 doe lost body weight and feed consumption values were 10 mglkglday dosage DDoGe28626af1teirn 1t4heda1i0lymdgo/skagg/edsa.y dAodsvaegresegrcloiunpicaalboorbtseedrvaantidownsasinscalcurdiefdicsecdaonnt feces (DGs 12, 16 0 20, and an ungroomed coat 23 and 24), no (DGs 24 to 26). feces (DG 13 to All lobes of the 15, 21, 22, 25 and 26) liver were mottled red and tan and the necropsy. This mucosal surface of the stomach doe aborted seven dead fetuses had and numerous black areas two dead fetuses were at in utero. The fetuses that were fetuses appeared normal for in utero had edematous heads their developmental ages. This and doe necks; all generally other lost weight and had reduced feed consumption values after DG 9. 000835 418-012:PAGE D8 418-012P PAGE 8 Doe 8262 in DG 22 after the 10 mg/kg/day 14 daily dosages. dosage group aborted and was sacrificed on Adverse clinical observations included a mass on the nose (DG 13). (DGs 7to 21), scant At necropsy, the cut feces (DGs 11 surface of the to 12, 14 to21) andnofeces small mass on the nose revealed a rsemmoaoitnhedpiinnkumtaetreor.ialT.hisNidnoeellaotsetrbeosodryptwieoingshtwearned afbeoerdtecdonasnudmpntoioconnvcaelputuessewse.re reduced after DG 9. Doe 8264 in DG 23 after the 10 mg/kg/day 14 daily dosages. dAodsvaegrseegcrloiunipcaalboobrsteerdvaantidownsasinscalcurdiefdicsecdaonnt feces (DGs 12, 14, 15, 19 red substance in the cage and pan 20), (DG no feces (DG 13, 16to 18, 21 and 22) 22). All tissues appeared normal at and a necropsy. Nine utero. This doe late lost resorptions were body weight after aborted and no DG 8 and feed conceptuses consumption remained in values were reduced after DG 9. Doe 8265 in DG 25 after the 10 mg/kg/day 14 daily dosages. dAodsvaegresegcrloiunipcaalboobrsteerdvaatnidonwsasinscalcurdiefdicsecdaonnt ifnectehse (caDgGes p9atno (20D,G2235)a. ndAl2l4t)i,ssnuoe fseacepspe(aDrGesd2n1oramnadl 2a2t)naecnrdopasyre.d Tshuibsstdaonece aborted one dead utero. All fetuses fetus and appeared five live fetuses normalfor their and one early developmental resorption were in ages. Body weight losses occurred after DG 8 and throughout the dosage period. feed consumption values were reduced 20mg/kg/daydosage dDaoiley8d2o6sa6giens.theA2d0vemrgskegcllidnaicyaldoobssaegrevagtrioounps wpraisorftooudnedadtehaidncolnudDedGs1c7anatffeerc1e1s (DGs 1210 perivaginal 15), a red substance in substance (DGs 16 and c1a7g)eapnadn,exncoesfescessa,livuantgiornoo(mDeGd17c)o.at, a red NNeeccrrooppssyy arlesvoearleevdeaalwehditteearfsroitnhythmeatleerftiadliaipnhtrhaegemastoipchalogbuesaannddiinntfhlaeteldeftlulnagtse.ral lobeofthe lungs (approximately 2.0 fundic mucosaofthe stomach (1.5 x cm 0.5 in length) and cm and 1.5 x two 0.5 eroded areas in cm). Ten early the argeesso.rptBioondsywweerieghitn luotesrso.occAlulrrfeedtuasfetserapDpGea8raedndnoitrsmfaeledfocrotnhesiurmdpetvieolnopvmaleunetsal cwoenrseidreerdeudceadntihnrtouubagthioountatchceiddeonstabgeecpaeurisoed.ofTthheetdeeaartihn otfhethliesftdlouengwalosbe; however, this doe had the doe was moribund. adverse clinical and necropsy observations that indicated 000836 418-012.PAGE D-9 418.012P.PAGE dDaoiley8d2o6sa7geins.theA2d0vemrgs/ekgcl/idnaicyaldoobssaegrevagtrioounps winacslufdoeudndscdaenatdfeocnesD(GD2G0s a9f,te1r013 alensdio1n6s wtoer19e),obnsoerfevceedsb(yDnGescr1o1ptsoy 1a5n)datnhdiretxeecnesdseasdalfiveattuisoens(wDeGre19i)n. utNeroo.groAsls foectcuusrersedapafpteearrDedG dosage period. n8oarmnadlffeoerdthceoinrsduemvpetlioopnmevnatlaulesagweesr.e Breodduyceweditghhrtoulgoshsoeust the 0D10o1ed5a)8i,l2ys6od8fotisonargtehlsieq.u2i0dAfmdegvc/eekrsgs/e(dDcaGlyisndi1coa2slaaognbedseg1rr3vo)a,utpiaowtnaasnsipnfecorluiunvdadegddinesaacladnstounbfseDtcaGensc1(e7D(aGfDstGe9r16) faindd ninotfheeceasbd(oDmGin1a6l). caNvietcyr,oapsfryiarbelveesatleodmaacphpraonxdimaatreulpytu1r8edmgLalolfbclladoduedry.grNeienne dead fetuses and one early resorption were in utero. All fetuses appeared naonrdmfaelefdorcothnesiurmdpetvieolnopvmaelnuteaslwaegrees.redBuocdeydwtehirgohutghlooustsetsheocdcousraregde after DG period. 9 DDoGe1892a6f9teirn 1t3heda2i0lymdgo/skagg/edsa.y dAodsvaegresegrcloiunipcaalboobrsteerdvaatnidonwsasinscalcurdiefdicsecdaonnt tcfohenecfecisram(gaeDtGipsoann1o1(ftDpoGer1s91i)9s),t.eenxOtctehasedsrvestrahslaienvacatlniinoioncba(lsDeorGbvssaet1riv8oanatinoodfnus1,n9)gnraoongodrmoaesdsrecldoeasstiuobansnstdwaencree in observed by necropsy. This doe aborted three dead fetuses and five live fetuses wweeirgehtinluotsesreos; oalclcfuertruesdesanadppfeeaerdecdonnosrummapltifoonr tvhaeliruedsevweelroepmreendtuacledagaefste.r DBoGdy8 Doe 8270 in the 20 mg/kg/day dosage group was found dead on DG 16 after nine daily dosages. The only adverse clinical observation prior to death was ssucrafnatcfeeocfesth(eDsGtsom9taoch15t)o.beNercerdoapnsdy driefvfuesael.edFtihvee nfoent-ugslesanadnudlaornmeuecaorslayl resorption were in utero. All fetuses appeared normal for theirdevelopmental waegerse.reBdoudcyedwetihgrhotuglhoosustesthoeccduorsraedgeafpteerrioDd.G 7 and feed consumption values A.2. ClinicalObservations 2D5o,s5a,ge1-0deapnednd2a0nmtg/inkcgr/edaasyesdoinsaagdveegrrsoeupcsl.inicOanl eobrsaebrbviattiinotnhseo2c.c5urdroesdaginetghreoup and all rabbits in the 5, 10 and 20 mg/kg/day dosage groups had scant feces (ifnirtshteob5,se1r0veadndon20DmGgs/k1g4,/d1a3y,d9oasnadge9,grreosuppescthivaedlyn)o. fOencee,s f(fiivrestaonbdsethrrveeed roanbbits 000837 418-012:PAGE D-10 418012P.PAGE 10 DG 18, 12 group had and 11, excess respectively). salivation (first Three rabbits in observed on DG the 20 mg/kg/day 17). Two rabbits dosage that aborted roredwesurbesftoaunncdedineatdheincaegaechpaonf t(hferst10obasnedrv2e0dmogn/kDgG/sda2y2daonsdag1e6,grreosuppescthiavedlya) Two rabbits in the 20 mg/kg/day substance (first observed on DG dosage group had 16). One rabbit in a red each or of tan the perivaginal 2.5, 5, 10 and 20 mg/kg/day 24,24 and dosage groups had 16, respectively). an ungroomed coat (first observed on DG 18, 23, bAlelcoatuhseer: cli1n)itcahleoibnsceirdveantcieosnswewreerenoctodnosisdaegree-ddeupnernedleatnetd;toort2h)ettheestoabrsteirclveations ocourred in (underside, only one rat. These clinical observations head, back and/or limbs) for 2, 1, 1, 0, 2, included localized alopecia 0 and 0 does in the seven grresopuepctdiovee tdhoastaagbeorgtreodu,pos,nearmaabbsist ionnetahcehnoofsteheof0.o1nean1d0 2m0g/mkgg//kdga/ydadyosdaogseage groups one 0.1 that had soft or liquid feces (first mg/kg/day dosage group rabbit observed on DG on one day of the 12) and study. scant feces in A3. Necropsy Observations sTchheerdeuwleerdesancorifniecec.ropAlslynoebcsreorpvsaytioobnssefrovratthieonrsabinbirtasbbtihtast tshuartvidvieeddournatiblorted were described previously. B. SMautmemranraileBsod- yTaWbeliegsh4tsanadnd5;BIonddiyviWdeuiaglhDtatCaha-nTgaebsle(F1i3gure 1; aRlalbrbeictosridnedthient1e0rvaanlsda2ft0ermgin/iktgia/tdioanyofdodsoasgaegeg;rothueprsehwaedrebondoyswuerivgivhitnglopsrseegsnfaonrt 5rambbgi/tksgiindtahyedsoesgargoeupgsroauftpesr hDaGdsd2e6craenadse1d9,broedsypewcteiivgehlty.gaiTnhseo2r.b5oadnydweight losses for the entire dosage postdosage period (DGs 21 period to 29), (calculated as DGs 7 to body weight gains were 21). During increased in the the o2c.5cuarnsdin5tmhge/skegt/ydpaeysdoofssatgudeiegsr.oupBso,dya wreeibgohutndgapihnsenfoormtehneoennttihraetgceostmamtoionnly (peDrGisod7(tDoG2s9)0wteor2e9)readnudcetdheienntthiere2g.e5satantdio5nmpge/rkigo/ddaafytedrotshaegienitgiraotiuopns.of dosing Body weights dosage of the and test body weight article. gains were unaffected by the 1 mg/kg/day 000838 418-012.PAGE D-11 418.012P PAGE 11 C. Ab(sSoulmumtarei(egs/d-aTya)balnedsR6elaantdiv7e: (Iqnldkigviidduaayl)DFaetead-CToanbsluem1p4t)ionValues aRbasbobliuttseian ntdhere1l0atainvedf2e0edmgc/okngs/udmapytidoonsavgaleuegsrofuorpsallhardecsoervdeerdeliyntreervdaulcseadfter initiation of dosage; there were no after DGs 26 and 19, respectively. surviving Absolute parnedgnraenltatriavebbfietesdincotnhsesuempgtrioounps tvhaelu2e.s5faonrdth5emegnt/ikrge/ddoasyadgoespaegreiogdro(ucpasl.culDautreidnagsthDeGpsos7ttdoo2s1a)gewepreeriroedd(uDcGeds i2n1 tsou.r2v9i)v,infgeegdrocuopnss,uamlptthioounghvatlhuee5s wmeg/rkegg/ednaeyradlolsyacgoempgarroaubplheaadmsolingghtltyheinfcirveeased feed consumption `commonly occurs values during in these types this period, of studies. a rebound phenomenon that Feed consumption values were reduced in periodafter the the 2.5 and 5 initiation mg/kg/day dosage groups of dosing (DGs 7 to 29) for the entire gestation A1bmsgo/lkugte/daanyddroeslaatgieveoffetehdectoesntsuarmtpictlei.on values were unaffected by the D. aCnades9;arIenadni-vSiedcuatlioDnaitnag -aTnadblLietste1r5tOhbsreoruvgahti1o7)ns (Summaries - Tables 8 rCaabebsiatrsewaint-hseocnteioonrimnogroebslievrevfaettiuosnesswienrteheba0,se0.d1,on1,52,.45, a4,nd55anmdg/2kgp/rdeagynadnotsage groups, respectively. Caesarean-sectioning There in the w1e0raenndo2p0remggn/akngt/draabybditosstahgaet survived groups. to DG 20 Fetal body twheeigphetrscewnetreresseovrebreedlycornecdeupcteudsaensdpetrhelinttuermwbeerreofinecarrelyasaendd(laaltoengrewsiotrhptthieo.ns and ocfontchoemfiotuarnptrdeegcnraenatsdeoiensliintetrhissizdeo)sian gtehegr5omugp/skugrlvdiavyeddotosaCgaeesgarrouepa.n-Only two gsercotuiponainsg.comFpetaarlebdotdoytwheeicgohntcsuwrerernetaclosnotrroeldsubcuetdwienrteheat2.t5hemlgo/wkegr/deanyddoofstahgee range observed historically at the Testing Facilty. aAldlmiontihsetrrCataieosnaroefatnh-esetecsttiaorntiinclge or at litter parameters dosages as high were as 5 unaffected mg/kg/day. by The litter caovmepraagreasblfeoracmoorpnograthleutfeiav,e implantations and surviving dosage percent groups. live No male does fetuses were hada litter consisting placentae aofppoenalyrerdesnoorrbmeadl.conceptuses, there were no dead fetuses and all 000839 418-012:PAGE D-12 E. | i ary - T: 10) Totals of 42, 37, 34, 42 and 11 fetuses were examined externally for gross alterations in the five respective dosages groups with live litters. Three fetuses had gross external alterations. One fetus (8251-1 0) in the 2.5 mg/kg/day dosage tghreou0p.1hmagd/akgc/ledfatysdnooustagaendgraobuspenhtadincfiestoarls.groTswsomafleftoursmeasti(o8n2s4;3-f8etaunsd82842344--87h)aidn a short snout, displaced and small nares and fetus 8244-7 had abdominal distention. IV. Recommendation: Based on these data, dosages of 0 (Vehicle), 0.1, 1.0, 2.5 and 3.75mg/kg/day PFOS were recommended for the developmental toxicity study in rabbits (Argus Protocol 418-012). The 0.1 mg/kg/day dosage is expected to be a no-observable-effect-level (NOEL) for both maternal and `embryo-fetal toxicity, and the 3.75 mg/kg/day dosage is expected to produce minimal maternaltoxicity and little or no developmentaltoxicity. Cost erm Alan M. Hoberman, Ph.D., DABT Date Director of Research Ean' ond G. York AasnsdocSitautdey DDiirreectcotror ns O6ame 99 . DABT Date search 000840 MATERNALFBigOurD1eY WEIGHTS || apdJl[. = - Lo gasefIlT e---- Lop Ce Ta en ses] | non EG i ge PR re w= | aN . See "rl A/ R =| Tra, 0 2 Sw Nx / - Ts] ssucraonr 11] o= a ee | |LI-- EL Shs ge hn hn nw |, .8 ig G5 Pog g ir meatsongor heen 5of preeend eevee 8 : g [I goE T EEe miEHyEEiEEi y .: {EmEERI Bo hieIh ouna5020 0aay ofSesemion | EREERNEE Pgiois3 {hr EEE Ss i8 ' Pio a LIA Bn 3S a 3 g a 3 3 i:  EERE i S wes memosowras 4 3 0 a Bp pmehen mddt pE ryeet ie on th ee bd 1 etn Ta wee vee a . E2 gE 5 P i s3 g8 :g 4 .& 2 28 iisf g Il rrr eri ae ae . f gw nce. aaa ELT arn ren 1 tain ane amen amsoa 83 FI [SE] FEL g A He Bg3 g 2 = Cn on a . oo ono . EF orn rs mien non oi gE 3 t E RrR .erE ese iioeo g So , 8 iE rn rir mieten amen dee S mim. LuLLInin Tmo s 2 REE .8 z :z io: J J----) worsen oinieon aasmsieem ann mes mien ie abit g mew sale lin SEg w TEmEa ... 8 1 EErERTAma 3 i: 38 : 2oz iLA oe g B EaEiuaen a vatue That sas Seveciaed withoption .i ioe inn man wi te am g PIE EERIE a3 3 .2 3 8 2 oo g8 Tinoommooamn ple ewe em g nL ie. g .g id8 LI omen le s S mup----_, . . . . sf & oT ise 3 ho 01 yt in prod sir yh i 5: 208 s mm " ame |8J i p--m--Te--L--onwlsaloen i i HiE 3 5 ar m-- ---- ---- aT wm ,B g8 Ii:3 LI s 3 8 LI  IRila - . g2 i. &8 82 Ei A g LEM EE .f Re : iBg g . 3 3 2 i3 ii 2 [I I 2 29 8 wm Poy EE BE ae EE TELLIN. 2 .E : Bog o TERE, gg i: , ifa g 8 FI Ii  # OEE ip i mumommmm 3 FW oREB TL T cove wee ote 1 or er shen stot sv, Lf3 :8 Pi g g LE memo * ET rots Soa a rnste te 1 os s rthe mfr re g igE 23 ii CRETE -- : os it g ,2 8 3 a F5 r38 . i, g& 3 ia gf @ a I 82 3it gg , 8 3 i Poe J g :f 3g HI ig S$ 3 338 .f igE :5 FI g8 g g8 Hii :i g :@ if 3g 85 HEE OE g S 2 5 2 g2 E g Z .f ig 8 i 5 a2 LI : i 8 [I 2 8FE 3 ig ,vidg& # id 88$s .5 tT 3 2 o i8 aShl ben" 514A TOHICETY ETWK oF HSE 1 maine i CTR PEARED en. E38MoT meme. :z iE i if [I wo - a . Ln noan 8% 3 HER 3 g io g iisd Pisf : tl TR 8 3: 8 Cd z3 3PR BOR | . Em rrmmaaseaassmmewaemmnne Li E8 BERL 33 RL pe es snr cee Bg og 418-012:PAGE D-66 418.012P:PAGE 66 ATTACHMENT 1 PROTOCOL 000894 418-012:PAGE D-67 418012P.PAGE 67 nesearcs, Argus Research Laboratories, nc. 4 rgus pod Ho9rT0s5rhSaihme,eesPhaytnnDeraiyviea,anrBiual.alda1i9ne0g4rA4 -------------------------------------------------------- PROTOCOL 418-012P `SPONSOR'S STUDY NUMBER: T-6295.10 STUDY TITLE: Oral (Stomach Tube) Dosage-Range Developmental Toxicity Studyof PFOS in Rabbits PURPOSE: TESTINGFACILITY: STUDYDIRECTOR: `SPONSOR: "The purpose of this study is to provide information for the `tosxeilceicttyio(neomfbrdyoos-aegteasltotoxbiecuysaenddintetrhaetodgeevneilcoppomteennttiaall) study of PFOS administered orally via stomach tube to New Zealand White [Hra:(NZW)SPF] presumed pregnant female rabbits. Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1287 Telephone: (215) 443-8710 Telefax: (215) 443-8587 ARsasyomcioantde GD.irYeocrtko,r oPfh.R0e.s,eDarAcBhT 3M Toxicology Services 3StM. PCaeunlt,erM,inBnueilsdoitnag 252104-42E--100200 STUDY MONITOR: Marvin T. Case, D.V.M., Ph.D. Telephone: (612) 733-5180 Telefax: (612) 733-1773 SATLUTDERYNMAOTNEITOR:~~ ATenlderpehwonMe.:Se(a6c1a2t), 5P7h.5D-.3161 Telefax: (612) 733-1773 000895 418-012.PAGE D-68 418.012P.PAGE 68 Protocol 418012P Page? REGULATORYCITATIONS HUaSm.onFiosoadtiaonnd;DGruuigdeAldimnieniosntrdaettieocnti(o1n99o4ft)o.xiIcnitteymtaotiroenparlodCuocntfieonrefnocremeodnicinal products. FederalRegister, September 22, 1984, Vol. 59, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58 fJoarpSaanfeestye SMitnuidsitersyoonf HDeraulgtsh,aMnHd WWelOfradriena(n19c9e7)N.umGboeord2L1,abMoarracthor2y6P,ra1c9t9i7.ce Standard European Economic Community (1888). Council decision on 28 July 1989 on the m`aacncdeapttiaonnceobnyctohmeplEiuarnocpeeawinthEcproinnocimpilcesoCofmgmouonditlyaobofraatnorOypErCacDtidceec.isOifofnic/iraelcJoomu-rnal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. REGULATORY COMPLIANCE: "rTehgiuslsattiuodnyswcililtebdeacboonvdeucintetdhaitntthheeTsepsititnogf FtahceiiGtoyopderLsaobnonrealtowirlyl aPrdahcetrieceto(GtLhPe)Standard OFpaceirlattyinQguaPlrtoycAedsusruersanfcorelUanbiotra(tQoArUy)opweilrlatniootnasudaintdtahetaproctoolcleoclt,iotnh.e rTahwedTaetsat,itnhge reports or the critical phasesofthe study. ADilrecchtaonrgaensdothereSvpisoinosnosro,ftdhaitsedpraontodcomlaisnhtaallibneeddowictuhmtehnetperdo,focsoilg.ned by the Study SCHEMATIC OF STUDYDESIGNANDSTUDY SCHEDULE: `See ATTACHMENT 1 to the protocol 000896 418-012:PAGE D-69 418-012PPAGE 60 Protocol 418P-a0g1e230 TA ESTRTIANC DVEL HICE LE: Identification: TestArticle: NPahmyseic:al Description: ~~ PLiFgOhSt-.colored powder. LotBatch Number. 217. PuSrpietciyf.ic Gravity: ~9088..9%. Expiration Date: May 2000. Information on the identity, composition, strength and purityofthe test article is on fle with the Sponsor. Vehicle: 0.5% Tween 80 in Reversed Osmosis Membrane Processed Deionized Water (R.0. Deionized Water). Supplier and lot identificationof Tween 80 to be documented in the raw data. tNoeibteheprrtehseenStpionntshoervneohrictlheetShtatudwyouDlidreicnttoerrfiesraewwairteh tohfearneysuplottseonfttiahliscosnttudaym.inTahnetrsefliokreel,y no analyses other than those mentioned in this protocol wil be conducted. Safety Precautions: Gloves, mask, appropriate eye protection and a uniformfiab coat are to be worn during f(oMrSmDulSa)tisonaptrteapcahreadtitoonthaendprdotooscaogle(aAdTmiTniAsCtrHaMtiEonN.T 2T)he Material Safety Data Sheet Storage: VBeuhlikcTleesCtoAmrptiocnlee:nts: PPrreeppaarreedd VFeohrimcullea:tions: RRoooomm tteemmppeerraattuurree.. RFrooozmente(-m2p0erCa)t.ure. AJlullitaenstGaurltbiicnlseksih,iMpamnenatgsetrotofheForTmeusltaitnigoFnasc,ialtttyhsehopurledviboeusaldydcrietsedseaddtdortehsse aatntdention of telephone number. `caSrhtiopnmsensthsosuhlodubled lianbcelluedde iapnpfroorpmraitaitoenlyc.onTcehrenirnecgipsiteonrtasgheocuolnddibteionnostfainedd sinhiapdpvinagnce of shipment. 000897 418-012.PAGE D-70 418012P.PAGE 70 Protocol 418.012P Pages EORMULATION: FreqoufPerepnaractioyn: Formulations (suspensions) will be prepared daaitlthye Testing Facility. Vehicle will be prepared weekly at the Testing Facilty. Detailed preparation procedures are attached to this protocol (ATTACHMENT 3). Adjustfomr Peurnitty The test article will be considered 100% pure for the purpose of dosage calculations. TestingFacilityReserve Samples: The Sponsor wil reserve a sample (1 g) of each lot of the bulk test article used during tthhee vceohuircsleeocfotmhipsonsetundtys. uTsheed TdeusrtiinnggtFhaeccilotuyrswielolfrtehsiesrvsetuadys.amSpalmepl(5esmLw)ilobfeeascthorelodt of under the previously cited conditions. ANALYSES: t`hSeamcpoluersseadodfittihoenasltutdoyt.hose described below may be takenif deemed necessary during Bulk Test ArticleSampling NInofoarnmaaltyisoensoonftthheesbtualbkiltteystofartthieclbeuwliklltbesetcaortnidculectiseodndfuirleinwgitthhethceouSrpsoensoofrt.his study. Analyses of Prepared Formulations: AcotntdhuecrteeqduedsutrionfgtthheeScpoounrssoer,ofntoheansatludyys.esHoofwperveepra,rerdectoersdtsarwtiilclbeefomramiunltaatiinonesd wtioll be document how the test article formulations were prepared. DISPOSITION: aPrrteicplaerweidllfboermrueltatuinoends twoiltlhbeeSdtiusdcyarMdeodnatiatthteotThreestpirnegviFoaucsiliytyc.iteAldlardedmraesisn.ing bulk test 000898 418-012:PAGE D-71 418012PPAGE 71 Protocol 41P8a-g0e1s2 TESTSYSTEM: `SpecianedRsea/soSn ftorrSelaecitionn: The New Zealand White (Hra:(NZW)SPF] rabbit was selected as the Test System tbhercoauugsheo:ut1t)hietiisndounsetrnyofno-rronodnecnltinmiacmalmastluidaienssopfedceiveselaocpcmeepntteadl atonxdicwiityde(leymbursyeod-fetal toxicityRteratogenicity) 2) this strain of rabbit has been demonstrated to be sensitive to adenvde4l)optmheentteaslt atrotxiicnlse;i3s)phhiasrtmoarcicoallogdiactaalalnydacetxipveeriinetnhceesepxeisctieast tahnedTsetsrtaiinn.g Facility"; NumabndeSerx: Population evaluated: 35 timed-pregnant female rabbits (5 per dosage group). BW odyeiganh dAgte: "The individual body weights of the female rabbits wil range from 2.5 kg to 5.5 kg; the rabbits wil be approximatelyfiveto seven months of ageatthe time of study assignment. Actual body weights recorded at receipt and at study assignment wil be documented in the raw data. Source: Covance Research Products, Inc. Swampbridge Road, Box 7200 Denver, Pennsylvania 17517 `The rabbits will be shipped in fitered cartons by truck from Covance Research Products, Inc., Denver. Pennsylvania, to the Testing Facilty. Identification: Rabbits are permanently identified using Monel seft-piercing ear tags (Gey Band and Tag Co. Inc., No. MSPT 20103). Female rabbits are given unique permanent identification numbers when assigned to the study on the basisofday 0 of presumed gestation body weighs. ANIMAL HUSBANDRY: Al cage sizes are in compliancewiththe Guideforthe Care and UseofLaboratory Animals 000899 418-012.PAGE D-72 418.012P.PAGE 72 Protocol 418P.a0g1e2s0 Housing: The rabbits will be individually housed inunitsof sitxo eight stainless steel cages. No nesting materials wil be supplied because the female rabbits will be sacrificed before parturition is expected. RTooemAmir, peraatnduHumridiety: `The animal room is independently supplied with at least ten changes perhour of 100% fresh air that has been passed through 99.97% HEPA fiters. Room temperature wil be. wmialilnatlasionbede maton6i1toFre(d16coCn)sttaont7l2yaFn(d2m2aiCn)taanidnemdoantit3o0r%etdoco7n0s%ta.ntly. Room humidity Light An automatically controlled 12-hour light 12-hour dark fluorescent light cycle will be: `maintained. Each dark period will begin at 1900 hours EST. Diet Abpeparvoaxiilmaabtleeltoy e15a0chg roafbbCietrteiaficehd dRaaybbuinttiChthoewfirs#t5d3a2y2of(PdMoIsaNguetr,itaitownhIincthertnaitmieonal) will a`Tphperocxeritmiaftieedlyfe1e8d0wigllobfethaevasialambelecefrrtoifmieidndfieveidduawlil,l sbteaionflfeesrsesdteteole, a"cJh-tyrpaebb"ifteeeadcerhsday. attached to each cage. Water: Water will be available ad itu from individual bottles attached to the cages or from an automatic watering access system. All water wil be from a local source and passed through a reverse osmosis membrane before use. Chiorine will be added to the pthraonce1s.s2epdpwmatcehrioarsinaeabtacttheeriotsitmaet;ofparnoacleyssisse.d Wwaatteerriisseaxnpaelcytzeeddtmoocnotnhtlayinfonropomsosribele bacterial contamination and twice annual for possible chemical contamination. Contaminants: Ne10itbheeprrtehseenStpionntshoercnerotriftiheed SditeutdoyrDiirtechteodrriisnakiwnagrweaotefraantylpeovteelnstitahlatcwonotualmdininatnetrsfelriekely pwietrhfotrhemerdesbuylttshoeftfheiesdsstuupdpyl.ieTrhoerretfhoorsee,mneontainoanleysdeisn otthihserprtohtaoncotlhowislel broeutcionnedluycted. 000900 418-012:PAGE D-73 418012PPAGE 73 Protocol 418P-a0g1e2?0 MRATAINNGDAONMDIZATION: Tshaemefesmoaulreceraabnbidtsstwriallinbbeenfaotruerasllhyibrpedmatetottnhhete STuepsptliinegrFbaycibltrye.edTehremadlaeyrmaabtbiintsg oofccthuers TwielsltbiengdFeasciiglntayteafdtedramyaOtionfg,prteo saurrmieved ognesdtaatyio1n.ofTphreesruabmbeidtsgweisltlabtieons.hipBpefeodrteo sthhiepment ogfestthaetiroabnbbitosd,ythweeigShutpsp.lieAr wciollmpfuotrewrar-dgebnreereadtiendg (rweeciogrhdts-oarndderdeady) Oraonfdopmriezsautmieond procedure will be used to assign the rabbits to dosage groups based on this information. ADMINISTRATION: RR outee andasfoo r Chon ice "The oral (stomach tube) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately administered; and 2) t is one of the possible routes of human exposure. Meat ndFh reqo uend cy: Female rabbits will be given the test article once daly on days 7 through 20 of aptreapspurmoexdimgaetsetlatyitohne. sDaomseagteimsewielalcbhedaadyj.usteddailyfor body weight changes and given Ratfoir DoosangeSaelelctieon: Dosages willbe selected by the Sponsor on the basis of previous studies conducted with the test article. 000901 418-012:PAGE D-74 418012P-PAGE 74 Protocol 418P.a0g1e2s0 DC osaogen Levcele s, ntratai ndVooln umess: [ DBoisgsegye ana | ConCcenetrsation| Voum9e gusBainNumver |1| [[5Towwm[o [10 [eswourscmsomven] [ ov |s[or[ oot [ to [sstwoncsommomven] [w|s[+ [or | 10 [sswoncconmnnven] [wv | 5s [ 2s [ om [ 10 [eswonroommonven] [vs[5 [ 0s | 10 [esmwourconmommen] [wv | 5[0[+ 1 0[esmwourronumven| (wow [5[= [2 Tw [ewsowrconmonven] The test aie vil be considered 100% purefo the pupose of dosage caluations TA ESTSN , ALANY DMES ASURE EMENS TS: Viability: All Periods: At least twice daily. ClOinicbal servandalorGteneiraloAppenarasnce Predosage Period: At least once. Dosage Period: Postdosage Period "Twice daily. Prior to dosage administration and once approximately one hour postdosage. Once daiy. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or Study Monitor. Body Weights: Predosage Period: Dosage Period: Day 0 of presumed gestation and on the dayofarrival at the Testing Facility. Dai. Postdosage Period: Daily. 000902 418-012:PAGE D-75 418.012P PAGE 75 Protocol 418P-a0g1e2sP EeedConsumptionValues: Predosage Period: Recorded daily after arrivalatthe Testing Facilty (values ot tabulated). Dosage Period: Recorded daily. Postdosage Period Recorded day. Feed consumption values during the dosage period will be tabulatedforthe same intervals as body weight evaluations. Caesarean-Sectioning Observations: Rabbits wil be Caesarean-sectioned on day 29of presumed gestation. The fetuses will be removed from the uterus and placed in individual containers. The rabbits will be `examined for number and distribution of: Corpora Lutea. Implantation Sites. d{aPltaac]entae that appear abnormal (ize, color or shape) will be noted in the raw Live and Dead (Alive fetus is Fetuses. defined as one that responds to stimui; a dead fetus is defined as daetaedrmfefteutsuessthdaetmodnosetsrantoitnrgesmpaornkdedtotosteimxutlrieamnedautthoatlyissisnoatremacroknesdildyeraeudtotloyzbee.d; late resorption.) E(aArcloynacnedptLuasteisRdeesfoirnpetdioanss.a late resorptionifitis grosslyevidentthat oarngeaanrolgyerneessoirpstihoans.)occurred; f this is not the case, the conceptus is identified as EetalObservations: `BodyWeights: `bTeheusbeoddytowdeiegthetromfineeaclihtefrefteutsalwibloldbyewreeicgohrdteadv.eraOgnelsy.body weightsof ive fetuses will 000903 418-012.PAGE D-76 418.012P.PAGE 76 Protocol 1P6a.g0e121P0 GrossExternalAlterations: fAeltfuesteussaelsswoilwlilbl ebeexeaxmaimniendedfofrogrrgorsosssexetxetrneamlalalatlertaetriaotniso.ns Ltaottehereesxotrepnttiopnosssainbdledebuatd sanuaclhysoebss.ervFaettiuosnesswwiillthnogtrboessienxctluedmeadlianlteeirtahteirodnastwailsubmemaprriezseartvieodn ionrnsetuattirsatlicbaluffered 10% formalin. All other fetuses will be discarded Representative photographsoffetal gross alterations will be taken. Sex: Al fetuses wil be examined internally to determine sex. MET OFSHACROIFID CE: Buseeudthtaonsaasciriafi-ceD rSapbebciitasl((vimaaninutfraacvteunroeudsbiynjeSccthieorn)inagn-dPliovuegfheAtnusiemsal(vHieaailntthr)apweirllitboeneal injection). NECROPSY: eGvraolsusatlieosnio(ncsowmiellspboenrdeitnaginteidssiunensewuitlrlableburfefteariended1f0r%omforrambabliitns ifnorthpeosvseihbilcelefuctounrterol dgirsocuapradtedt;hethdeissecraertieocnoomfmtohen,StsupdoyntDiarnecetoour)s. le(sEixocnespitniorna:bbiPtsa.r)ovUanrlieansscyssptecsiwfiilclalbley cited below, all other tissues will be discarded. ScheduledSacrifice: Ogrnosdsayne2c9roofppsryoefsuthmeedthogreasctiact,ioanb,dfoemmianlaelraanbdbitpselwviilcl vbiescCeareasawrielabne-speecrtfioormneedd., aUntderai of aapbpsaernecnetloyfniomnpplarnetgantainotn sdioteess.will be stained with 10% ammonium sulfide to confirm the Rabbits FoundDead orMoribund: dRealbibvietrsy wtihlaltbdeieeoxramarienesdacrfoirfitcheed cbaeucsaeuosfedoefamtohriorbumnodricboundnidticoonn,diatbioorntioonn tohrepdraeymatthuere ofebtsuesrevsatainodn/oisrmdaedleiv.erePdrepgunpasnwciyllsbteateuxsaamnidneudtetrointehceoenxtteennttspwoisllsibbeler,ecuosridnegd.theAsboarmteed mwietthh1od0s%daemscmroinbiedumforsufleftiudseest.o coUtnefriiromftahpepaarbesnetnlcyenoofnipmrpelgannatnattidoonessitewisl'l.be stained 000904 418-012:PAGE D-77 418.012 PAGE 77 Protocol 41P6a.g0e12101 STATISTICALEVALUATION: Aapvperroaprgieastea.ndAdpdeirtcieonnatlagpersocwieldlubreescaalncdullaotreda.nalLyitsteesr vmaalyuebsewiplelrbfeorumseeddifwdheereemed appropriate. [DATAACQUISITION,VERIFICATIONANDSTORAGE: DDiarteactwoirllabned/hoarnadp-praonpdr/ioartceommapuntaegre-mreecnotrdpeedr.soRneneclorwdisthwiinll2b1edraeyvsiaefwteedr gbeynetrhaetiSotnu.dyAll boreigbionaulnrdecaonrddsinwdiellxebde. stAorceodpiynotfheallarracwhidvaetsaofwitlhlebTeesstuipnpgliFeadctiolttyh.e ASlpoornisgoinralupdoatna will yreeqaureasftt.erPmraeisleirnvgeodfttihsesdureasftwiflilnablersetpoorrte,d aafttetrhewhTiecshtitnigmeFatchielSypaotnnsoorcwhiallrgbeefcoornotnaected to determine the disposition of these materials. REC TOO BEMR AIND TAINS ED: PTreosttoAcrotliclaen,dVAemhiecnldemeanndt/so.r Reagent Receipt, Preparation and Use. ARnainmdaolmiAzcaqutiisoitnioSnc.hedules. Veterinarian Examination. TMraetaintgmeHnitst(oriyp.rescribed byStaff Veterinarian). GCleinneicraallOCbosmermveanttiso.ns and/or General Appearance. BFeoeddy WCeoingshutsm.ption Values. Caesarean-Sectioning and Fetal Observations. GOrrogsasn NWeecirgohpssy(iOfbrseeqruviarteido)ns. PSthuodtyogMraaipnhtse(niaf nrecqeui(rreodo).m and environmental records). PFaecekdinagndanWda/toerrSAhniaplmyesnest.Lists. 000905 418-012.PAGE D-78 418.012P-PAGE 78 Protocol 41P8a.g0e121P2 KPEEYRSONNEL: DiErxeecctuotriovefDiRreescetaorrcohf:RAelsaenarMc.h:HoMbielrdmreadn,S.PhC.hDi.s,tiDanA,BPTh.D., ATS ADsirseocctioartoefDLiarbecotroartoofryROepseeraartcihonasn:dJSothundyF.DiBraemcettort:, BR.aS.ymond G. York, Ph.D., DABT MMaannaaggeerrooffSAtnuidmyalCoOopredriantaitioonns:anVdalMeerimebAe.r,ShIanrsptietru.tiMo.naSl. Animal Care and MaUnsaegeCromomfitRteegeu:latDoreynaCoCm.plLieabnoc,e:V.MK.aD.thleen A. Moran, M.S. Consultant, Veterinary Pathology: W. Ray Brown, DV.M., Ph.D. ACVP REPORT: imlmeetdtieratreeployrtfoflolrotwihnegpcuormppolseetioofndoofstahgeens-eIlfeectpihoansfeo.r the full study will be prepared Aressuultmsmaanrdy croenpcolrutsiwoinl; betapblreepoafrceodntiennctlsu;dicngo:pyaobfsttrhacet,prsoutomcmoalr;iaemseonfdmtehentmse;thsoudsm.mary and individual tables; and reports of supporting data (if appropriate). The report will be tihnecldurdaefdt arespoarnt aapnpdentdwioxctoopitehseoffulthsetufidnyalrerpeoprotr.t.The Sponsar will receive one copy of INSTITUTIONALANIMAL CAREANDUSECOMMITTEE STAEMN: TInhsetitpurtoiocneadluArneismdaelsCcrairbeedanidn tUhsisepCroomtomciotltehea.veAbleepnrorceevdiuerweesddbeysctrhiebeTdesitnitnhgisFapcrioittoyc'osl tdhiastcoimnfvoorltv,edsitsutdryesasnoirmaplasinwitolltbheecaonnidmuact.ed in a manner to avoid or minimize The Sponsor's signature below documents the fact that information concerning the. nduepcleiscsaittiyvefosrtucdoyndmucatyinbgetohibstasitnueddy farnodm tthhee fSapcotntshoart.thiNsoisanltoetranantiuvnene(icnesvistarro)ily procedures were available for meeting the stated purposesofthe study. 000906 418-012:PAGE D-79 418.012PPAGE 79 Protocol 41P8a.g0e12103 REFERENCES: 1. oCfhrtihsetitaenr,atM.oSg.e,niHcopboeternmtainal,oAfMs.usapnednsSimointsh,ofTt.nHiFt.ro(f1i9u6o2)r.enoDnoes(aTgNeF-)range study `administered orally toNewZealand White rabbits. Toxicologist 2(1):40 (#143). 2. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of nSacliternecxeosn,eN(oPvreomcebeedrin7,gs1o9f8N3a)l.tJr. eCxiionn.ePSsyycmhpioats.iu4m5(,9)N:e7-w10Y.ork Academy of 3. FMeSu.ss(n1e9r9,2).ELLA.dLeicghatdkeepo.frGa.bE.b.itHfeenrntieistyy,datRa.:A.StHuodbyeromfahnis,toAriMc.al aconndtrCohlriasntiimaanl,s. Teratology 46(4):348-365. 4. Institute of Laboratory Animal Resources (1996). Guidefor the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. SImaplleawnstaktiioE.nss(t1e9l6i4e)n. aFmarUbteemreusthdoedreRaztutem. maArkcrho.skPoatphiosl.chEexnp.NaPchhawremiaskovlon 247:367. 000907 PROTOCOLAPPROVAL: FOR THE TESTING FACILITY Associraalee. DDieraercitoovreo,f PRhe.sDe.a,rDcAh BT rad Z, AsSBCiantdeDGir.eYcotrokr,oPfhD /DRAkeaBrchT Study Director DIY MDeemnbaeCr.,LeInbsoti,tuVt.iMon.aDl.Animal Care and Use Committee FOR THE SPONSOR lTa ire s Marvin T. Case, D.V.M. Ph.D. Study Monitor 418-012:PAGE D-80 418.012P PAGE 80 Protocol 4P1a8g-0e12104 [2=q0 TE. Date 12-900 -9 Date 17 Mig 28 pate >// 1S0 ater Date 000908 418-012:PAGE D-81 418-012P:PAGE 81 ATTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE 000909 ATTACHMENT 1 418-012.PAGE D-82 418-012P:PAGE 82 ProtocoPl4a1g8e-01t2ot2 STUDYSCHEMATIC DDOSAGE-RANGE DEVELOPMENTAL TOXICITY STUDY" Timed. Pregnant Rabbits Start of Dosage CaesareanSectioning' End of Dosage Day0 of Presumed Gestation Day7 of Presumed Gestation Day20 of Presumed Gestation PDraeys2u9meofd Gestation SE Dosage Period. a. For additional details see "Tests, Analyses and Measurements" section of the b. pFreottaolcoelv.aluations (all fetuses - external examinations). 000910 ATTACHMENT 1 418-012.PAGE D-63 418.012P PAGE 83 ProcolPa4g1e8.20011220 264UN 98 02UL98-15.4UL 88 260008 3tauLes 080CT 98 SCHEDULE Animals Arrive- Accimation Begins. DosagePeriod (D7athyrousgh 20of presumed gestation). Caesarean-Sectioning Period (Day 20 of presumed gestation). Letter Report `Summary Report a. The study initiation date is the date the Study Director signs the protocol. 000911 418-012:PAGE D-84 418-012P:PAGE 84 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 000912 418-012:PAGE D-85 MDAATTEARISAHLEETSAFETY -3M Center 5St5.144P-au1l0,00Minnesota 1-800-364-3577 or (612) 737-6501 (24 hours) Copyright, ALL rights r19e9s8e,rveMdi.nnesCootpayiMnignianngd/oarnddMoawnnluofaadcitnugrinogf tChoimspany. iisnfoarlmlaotwieodn pfroorvitdheed tphuartp:ose of properly utilizing 3M products 1) ptrheiorinafgorremeamteinotn iiss ocobptiaeidnedinfrfoumllSMW,ithandno changes unless 2) neither the distributed copy with nor the itnhteenotriiogninaolf iesarnriensgolda oprrofoitthertwhiesreeon TDIRVAIDSEIONNA:ME:3MCHEMICALS FC-95 FLUORAD Brand 10 NUMBER/U.P.C. Fluorachemical Surfactant 9988--00221017--00818083--57 0000--5511113355--0099306524--17 9986--00221017--30911064--15 IS2SZUFE-D0:00J2a-n1u0a44r-y1 29, 1- 998 So DSOUCPUEMRESNETD:ES:10N-3o7v9e6m-b9e.r 05, 1997 0000--5511113355--0029301815-.28 418-012P:PAGE 85 PFOS 1. INGREDIENT C.A.S. NO. PERCENT PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SSUULLFFOONNAATTEE............ 3278957.139--939-6382 --886 POTASSIUM POTASSIUM PPEERRFFLLUUOORROOAALLKKYYLL SULFONATE...... SULFONATE...... 20420-49-3 60270-85-5 3 2 -7 -6 POTASSIUM PERFLUOROALKYL SULFONATE...... ~3872-25.1 1 3 2. PHYSICAL DATA VBAOPIOLRINGPREPSOSIUNRTE::..................v.u.v.u.e.n.n.s NA NIA EVAVPAOPRORDAETNISOINTRYA:T.E.:..............0.00.0.0.0.| NA N/A SSPOELCUIBFIILCITYGRAINVIWTAY:T.E.R.:..................... csal.igh0t.6 Waters! PERCENT VOLATILE:.............. 0 (%Bulk) VISCOSITY:...........ccceeirNrIe(Ds0.1% Aqueous) MELTINGPOINT: .....0000 00000000 NID APPLEiAgRhtANCcEoloArNeDd,ODOfRr:ee flowing powder. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 000913 418-012.PAGE D-86 WJaSnSu:aryFC-299,5 1F9L8U8ORAD Brand Fluorochenical Surfactant 418012P-PAGE 86. PAGE 2 3. FIRE AID EXPLOSION HAZARD DATA FFLLAASWHMPBOLIENTL:INcTTSoLcE:o1o111n WHOIMAO AFULTAOMIMGANBILTEIOLNIMTIETWSPE-URAETLU:R.E:111.01.1.]. NWIIAA EXWTaItNeGrU,ISHCIaNrbGonMEDdIiAo:xide, Dry chemical, Foss SPEWPeCoaIsriAtLifuvFleIlREpprreFsoIstGueHrcTetIiNvGoerPcpRlrOoeCtEshDsiUunRrgEe,S:dienmcslnuddibnrgeathhelisnegt,appsaerlaft-ucso,ntabiunnekde,r coat P2nrdotepacnttisv,e cbaonvdesrinagroufnodr earxapso,sedWaiasrteasandofletghse, hefaadc.e sask, and UNU`SSeUeALHazFaIrREdouAsNDDeEcXoPmLpOSoIsOiNtiHoAnZAsReDcSt:ion for products of combustion. "4. REACTIVITY DATA STABILITY: Stable INNCoOtM.PAaTpIplBiIcLaIbTlYe.- MATERIALS/CONDITIONS TO AVOID: HAZARDOUS POLYNERIZATION: Hazardous polymerization will not occur. HAZGAarRbDoOnUSMoDnEoCxOiMdPeOSIaTndIONCarPbRoODnUCDTiSo:xide, Oxides of Sulfur, Hydrogen Fluoride, Toxic Vapors, Gases or Particulates. 5. ENVIROWMENTAL INFORMATION SPIOLbLserRvEeSPOpNrSaEcautions from other sections. Vacuum, use Wet sweeping cboempanounidgniortioMnatesrourtcoe.avoiCdleaGnustuipngr.esCiAdUuTeIOwNi!thAwavtaecru.um cPlleaacenerincaounld approved metal container. Seal the container. REDCoOMnEoNtDErDelDeIaSsPeOStAoL:Waterways or sewer. Do AGE Use in products or pSr7o70cesosfesthethaltowecsotuldECSr0esuolrtLiCnS0aCqounacteicntrcaotnicoenn.tratIinocnisnergarteeateirn tanhan miantdeursitarli.al CorombcuosmtmieornciaplrofdaucctislitwyillinitnhceludpereWsFe.nceDoifsp3osaclombustible Alternative: Dispose of Haste product in a facility peraitted to Abbreviations: N/D - Not Determined N/A - Not Applicable GA - Approximately 000914 418-012.PAGE D-87 JWaSnSu:aryFG-299,6 F1L99U8ORAD Brand Fluorocheaical Surfactant 418.012PPAGE 87 Pace 3 5. EWIROWENTAL INFORMATION (continued) accept chemical waste. E`NGVB6IlRunOerNg.MiElNlATaAuSLautnifDcAiTsAhF:i(sLhepoLnCSiDs, asFcartohoehadirWuisn)n-ogus(Pmigs/Le,phaRlaeisnbopwrosTreolusts()sS3a8lenmg/l, OGlaai;ran8e0r0s2)0=1=1Nimlg./L; 48-Hr. ECS, Daphnia Magna = S0 mg/L; COD=.004 REGVUoLlAaTtOiRlYe OIrNgFaOnRMiAcTICOoNm:pounds: N/A. Voc Less HZ 8 Exempt Solvents: WIA. DSienfcoereredgiusploastailo.ns Uv.aSr.y, EcPoAnsHualztardaopupslicaasbtlee NreugsubLesrti=onsMoonre a(uNotthorUi.St.ies EPA Hazardous). TTShCiAs, pErIoNuEcCtS,cCoOsSpLl,issAICWSi,thMItThIe acnhdemKiocraela.registration requirements of EPFCIRRAE HHAAZZAARRDD: CLNAoSS:PRESSURE: Mo REACTIVITY: No AGUTE: Yes CNAONIC: Yes 6. SUGGESTED FIRST AID EYETaCnOeNdTiAaCtTe:ly flush eyes minutes. Get immediate Wmietdhicallargsetteanmtoiuonnt.s of water for at least 15 SKITNaneCgONiTaAtCeTl:y flush skin contaminated clothing. wIifthirrliatragteioanmoupnetrssisotfs,watcearl.l Raempohvyesician. Mash Contarinated clothing before reuse. IN"AItLASTiIgOnNs:syaptons occur, remove person to fresh air. If signs,sysptons continue, call a physician. IFDrSiunLkoutewoD:glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION ev`eAvPoRiOdTEeCyTeIOcNo:ntact. Wear vented goggles. Abbreviations: ND - Not Deterained N/A - Not Applictble OA . Approximately 000915 418-012:PAGE D-88 JSaDnSu:aryFC-299,5 F1L9U9O8RAD Brand Fluorochesical Surfactant 7. PRECAUTIONARY INFORMATION (continued) 418.012P-PAGE 88 PAGE SK`IANvoPiRdOTsEkCiTnIOcNo:ntact. material. A pair of gWleoavresapspardoeprfisrtoe tghleovefsollwhoewninghanmdatleirnigalt(hsi)s are Fpeecrosmomneanlcepdr:otecbtuitoynl riutbebmesr.as Unseecesosnaeryortoo rperevofenttheskfinollcoownitnacgt: head bSeoveardieng,ofcoevietrhaelrlso,f tPherotfeocltliovweinggarmsaetnertisal(so:ther than gloves) should polyethylenepolyvinylidene chloride (Saranex). RECUsOeMMEwNitDhEDaVpEpNrToIpLrAiTsItOeN:local exhaust ventilation. Use in a well Voemnitsisliaotnesdbaerleoaw. rePcroomvsiednededsufefxipcoiseunrte vliemnittisl.atioInf teoxhamuasitntavienntilation 16 not adequate, use appropriate respiratory protection. RES"APvIoRiAdTObRrYeaPtRhOiTnEgCToIfONd:ust. Select one of the eapirators based on airborne concentration offollcoownitnagminNaInOtSsH aanpdproivned hAaclcformdaasnkcesWuiptphlieOdSHAairregruelsaptiiroantso:r, fhualllf--fmaacsek dduusstt aanndd mmiisstt respirator, respirator, fullface supplied air respirator. PR`EDVoENnTotIONeatO,F dArCiCnIkDENoTrALseoIkNeGESwThIeOnN:using areas thoroughly With sosp and Water. tWhiasshphraodnudcst.afWtaershheanxdploisnegd and before eating. RECKOeMepMENcDoEnDtaiSTnOeRrAGdEr:y. Keep container closed when not in use. FIRNEonfAlNaDmaEaXbPlLeO.SION AVOIDANCE: OTHNEoRsPmoRkEiCnAgU:TIOSNmAoRkYingINwFhOiRlMeATIuOsNi:ng this product can result in cofonttahmeinhaatziaorndouosf tdheecotmopboascictoionandp/roorducstssokemenatndionleeaddintostehcetifoonrm4atoifon this MSDS. WMS MAZARD RATINGS: PMEEARLSTOHN:AL2PROFTLEACMTMIAOBNI:LITXY:(Se0e pRErAeCcTaIuVtIiToYn:s, 0section 7.) EXPOSURE LIMITS INGREDIENT PPOOTTAASSSSIIUUMM PPEERRFFLLUUCORROOAALLKKYYLL SSUULLFFOONNAATTEE...... PPOOTTAASSSSIIUUMM PPEERRFFLLUUORROOAALLKKYYLL SSUULLFFOONNAATTEE...... VALUE NIT 00..11 MWGG//MM3 00..11 WMGG//WM33 TYPE AUTH SKIN" TTWWAA 3314 Y TTWWAA 334M YY Abbreviations: N/D - Not Deterained N/A - Not Applicable CA - Approximately 000916 418-012:PAGE D-89 418.012P PAGE 89 MJSaDnSu:aryFG-299,6 F19L9U8ORAD Brand Fluorochesical Surfactant Page 5 EXPOSURE LIMITS (continued) INGREDIENT ALE oI TYE AUTH SKIN POTASSIUM PERFLUGROALKYL SULFONATE... 0.1 WG/M3 THA am ThneeS.lKuIpaNoitneNgnOtTAimTauIlcOoNuc:sontmrLeiiabsbutraetndieonsuabtnsodtaetnyhece,esoevieitrnhadelirlcatebeyxdpoaswiuirrtbehornb'eyY' toruh,endecmrourteaSKnIepNoaurstriecfrueolruatretloy, By direct contact with the substance. Venicles can alter skin absorption. S2O3UhR:CE OFaEmXPROeScUoRmEmenLdIeMIdTEDxApToAs:ure Guidelines 5. HEALTH HAZARD DATA : EYEM3C4ONTEAyCeT:irritation: signs/sysptoms can include redness, swelling, pain, and tearing. SKIMSNiilgdCnOsNSsTkyAimCnpTt:oImrsritactanioninc(laufdteerrpedrnoelsosn,gedswoerllirnegp,eataendd ciotncthaicntg): eMaxytenbdeedabstoinreb.ed through the skin and persist in the body for an INMHaAyLATbIeONh:armful if inhaled. tMaiynebe absorbed by inhalation and persist in the body for an extended Single overexposure, above recomended guidelines, may cause: sIorrreinteastsionof (tuhpepernosreespainrdattohrryo)a:t, sicgonusg/hsinygsptaondms scnaeneziinngc.lude IF ISnWgAeLsLtOiWoEnD:is not a likely route of exposure to this product. tIhlilsnesmsatemraiyalr.esult from a single swallowing of a moderate quantity of May be harmful if swallowed. MUTMAuGtEaNgIeCnIiTcYi:ty assays indicate the product is not mutagenic. Abbreviations: N/D - Not Deterained W/A - Not Applicable CA - Approximately 418-012:PAGE D-90 MJSaDnSu:aryFG-299,6 F19L8U8ORAD Brand Fluorocheaical Surfactant 418012PPAGE 90 Pace "al WEALTH WAZARD DATA (continued) REPWRoOtDUtCeTrIaVtEo/geDnEiVcELOiPnHtEhNeTALraTtOXaItNSo:ral doses below maternally Toxic Tovels. OTTHChEaiRlsifHoPErrAnLoiTdaHuctWPArZ4oAspRoDs.niottIiNokFnnOoRwM8n5A.TITOoN:contain any substances regulated under A Product Taxicity Summary Shest is available. SECTION CHANGE DATES HeAoIG SECTION CHANGED SINCE November 05, 1997 ISSUE Sobreviations: WD - Wot Determined WIA - Not Applicable GA . Approximately TDIehN"ePLcIoiErnDrf,eocrtmIaNtaCiLoUnDofINiGtn,hethBiUdsTateNMOaTtiesrsLiuIeaMdlI.TSEaDf3eT%Ot,yWAKADENaStYaNIOSWhPWeLAeIRtERDA(NWMTASIRDESRS)A,NTisYEXPObFReElSiSevEeDdORto HnPEEeRRtGFhHOeARrNMTAANtBChIeELIOaRTMYpUOSrRAo'GdEuFcItOTFNEiSTsSRAFDLFETO.RoArUsPeA&rRTpIaiGsrUtLirAceRuslpaPorUnRsFipObuSlrEeposfOeRorCaOdnUdeRtSesErumiOitFanbilneg for uUCansniearuasetlaymcettwhiottdhheinofusteuhseeaundosrearpasbpiplikicncaoatwtilioeondng.eofa3GndivIeMcnonpttrrohadelu,cvta,riitestoiysmeoefosfsefnWathciitaoclrhsatrhteahtat TPhaertiucsuerlarevapluuraptoesethaendSHsupirtoabdluectftoor duesteerrsminmeethWohdethoferuiste iosr Taiptplifocratiaon. MDinu'e'perrtoro.ovritsdh,ees rosemmniosftsoeiroanpssotisosonirbiianlliteetlryeatctithoranotsniecilnecftotrhriaosniacsinfatorrasanessrtvfiieocrne,satyoSHhimatsavekecsursetnsooumletresd. rienpfroersmeanttiaotnioonbstaaisnedtofirtosn caomdpalteabtaesneessmayornoatccubreac8ys. curIrne'natddiatsiotnh,e information in the HSOS available directly from 000918 418-012:PAGE D-91 418-012P:PAGE 91 ATTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE 000919 418-012:PAGE D-92 418012P-PAGE 52 ATTACHMENT 3 Version: 41Pr8oto-c(o1l024J11U8-N2091P280) TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE Page 1013 Test Article: PFOS Vehicle: 0.5% Tween 80, in RO. Water "A. Purpose: The purposeofthis procedure is to providae method for the preparation orfabdboitssaogneAsrugsupsenSstiuodnys4o1f8P-0F1O2SP.and thevehiclefor oral administration to B. General Information: 1. All suspension containers wil be labeled and color coded. Each label will specify the protocol number, test article identification, Argus batch number, concentration, dosage level, preparation date, expiration date and storage conditions. 2a. Suspensions wil be prepared: X_ Daiy _ Weeky _For__daysofuse 2b. Vehicle wil be prepared: -- Daily X_ Weekly _For__daysofuse 3. Suspensions wil be prepared at a final dosage volume of 10 mLikg. 4. Xsa_fety:Gloves, lab coat, goggles or safety glasses and faceshield X_C DHuasltt--MFiascte RReessppiirraattoorr Z Full-Face Respirator/Positive Pressure Hood Tyvek SuitiApron 5. Dosage solutions adjusted for Free base and % Pury. -- Yes X_ No(Calculationsbased on 100%) __ FreeBase __ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol 000920 418-012:PAGE D-93 418012P PAGE 83 ATTACHMENT 3 Version: 41Pr8oto-c(o1l024J11U8.N2019P28P) `TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE Page 2013 NOTE: tTheestloarwtidcolseawgilel.beOnprceepathreedfianaslavoselruimalesdialurteioanchfireovmetdh,setihribgahrdsosaargeetotobe `adamdidneidsttoratthioen.containers; mixing should occur during sampling andlor C. Preparationof Vehicle 1. Aldadbetlhedecroenqtuaiinreerd.amHoeuantttohfewRa.tOe.rdetoio5n0izCe,d w5atCe,r taodadnthaepprreoqpuriiraetdely `CaAmLoCuUntLAofTITOwNeSe)n. 80 and mix until uniform (See TEST ARTICLE D. Test Article Suspension Preparation: 1. Tofotpesrtepaartriectleh(eS2e-emTg/EmSLT. Group Vil ARTICLE CsAusLpCeUnLsiAoTnI,OaNdSd)thinetoreaqnuired amount Vaephpircolperiaantdelhyeasitztehd,e lmaibxetluerdectoont8a0ineCr,. 4A5dCd ftohrearpepqruoixriemdataemloyu3n0t mofinutes. 2. Othnecree itshae tveissitbalretivcolretehxa,stdhiissswoilllveadc;hisepvientwhheildeetshieresdoleumtuiiosniocno.olsT.hi(sBmeasyure be prepared the day before use.) 3. `Taomopurnetpaorfestthoeck1s-umsgp/emnLs,ioGnro(uGproVuIpsVuIsIp)e(nSseioenT,rEeSmTovAeRTtIhCeLrEequired CALCULATIONS), add the required amount of vehicle and mix. 4. aTomporuenptaorfestthoeck.s5u-smpge/nmsL.ioGnr(oGurpoVupsVuIs)pe(nSseieonT,ESreTmAoRvTeItChLeErequired CALCULATIONS), add the required amount of vehicle and mix. 5. T`aomporuenptaorfesttohcek.s2u5s-mpge/nmsLi,onGr(GoruopupIVVs)u(spSeenesTioEnS,TrAeRmTovIeCLthEe required CALCULATIONS), add the required amountofvehicle and mix. 000921 418-012:PAGE D-94 418.012P:PAGE 94 ATTACHMENT 3 Version: 41Pr8oto-cLo1lP02a4Jg11eU8.N230o19Pf2839) TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE 6. `Taomporuenptaorfestthoeck0s.u1s-mpgeinmsLi,onG(rGoruopupIllIsVu)s(pSeenesiTonE,STreAmRoTvIeCtLhEe required CALCULATIONS), add the required amooufvenhictie and mix. 4 7. Writen by: aTomporuenptaorfestthoeck0s.u0s1p-emngsiimoLn, (GGrroouupplIlls)u(sSpeenesTioEnS,TrAeRmTovIeCLthEe required CALCULATIONS), add the required amoofuvenhictle and mix. rg Lo Approved by: |Z. Date: /2-guw-38 Clarification: 7No __ Yes (See attached clarification form.) ntiatsiDate: (hmbten RZ o-- 000922 APPENDIX E HISTORICAL CONTROL DATA 000923 418-012:PAGE E-1 SUMMARY OF REPRODUCTIVE INDICES NZW RABBIT PERIOD: JANUARY 19- JA9NUA6RY 1998 NUMBER OF STUDIES: 66 NUMBER OF RABBITS: TESTED 901 FOPURNEDGDNEAANDT 8661 DELIVERED PREAMABTOURRTEELDY 24 NUMBER OF RABBITS PREGNANT AT CAESAREAN-SECTIONING ON DAY 29 OF GESTATION: 825 NSUIMNBGELRE COOFNRCAEBPBTITUSSWLIITTTHER: RESORLBIEVED 11 ABORTED 2 MEANOr% % PREGNANT 9%0 AVERAGE # CORPORA LUTEA 87 AVERAGE # IMPLANTATIONS 89 AVERAGE LITTER SIZE AVERAGE #LIVE FETUSES 84 AVERAGE # DEAD FETUSES 00 AVERAGE # RESORPTIONS 04 AVERAGE# EARLY RESORPTIONS 03 AVERAGE# LATE RESORPTIONS or * Three were moribund sacrifices, one was attributed to an intubation accident RAMNEGAEN/OSrT%UDY (750-100) (8117) (38108) (32104) (01) (032) (028) (12) 000924 418-012:PAGE E-2 SUMMARY OF REPRODUCTIVE INDICES NZW RABBIT MEANOr% RMAENAGNE/oSrT%UDY ARVEESROARGPETI%ODNOSES WITH ANY 68 (0100) ACVOENRCAEGPET%USDEOSESREWSIOTHRBAELDL 0s (0200) AMVOERRAEGLEIV%E DFEOTEUSSWEISTH ONE OR 4 (800-100) A(V%EMRAALGEESISLEIXTTREART)IO, 512 (314610) AVERAGE FETALBODY WEIGHT (6) ~~ 4374 (31.85-55.74) AVERAGE FOR MALES (G) 426 (20555697) AVERAGE FOR FEMALES (G) 4308 (32255376) AVERAGE % DEAD OR RESORBED CONCEPTUSESILITTER 44 (0188) 000925 418-012.PAGE E-3 `SUMMARY OF MATERNAL NECROPSY NZWRABBITS OBSERVATIONS TPOETRAILOD# STUDIJEASNUARY 1996 - JANUARY 19788 TOTAL #DOES #PREGNANT 987261% ##ADIBEODRTED 2& ##DDEOLEISVEWRITEHD 1P0R0E%MARTESUORREPLTYIONS. 45 OFP0RsEGNANT 0244 05 EXTERNAL OBSERVATIONS Fecalmaterial in Lopcearliiaznealdraelgoipoencia Leftear,tom GROSS LESIONS THYMUS `Smal LUNGS Discolored Tein raightdriapMhurltaigplmealteisiclonosbe THORACI`CCCoAnVtIaiTnYedred fid LIVER Pale andlor discolored Acpcaetnttemuaotnedlloobbeusl)ar BACK Breakpresentin lumbarregion of spine RANGE /STUDY NO% ON % 210012 0011 (050) (050) 10m 01 (050) 1 011 01 (050) 6 065 04 (0200) 11001m 0011 ((004402)) 10m ot (040) 7 076 04 (0-160) 1 01 01 (050) 1011 01 (050) + aTthtrreibeutweedrteo maonriinbtuunbdatsiaocnriafcicciedse,notne was 000926 418-012.PAGE E4 SUMMARY OF MNAZTWERRNAABLBINTESCROPSY OBSERVATIONS GROSS LESIONS BACK (CODNoTr)salmuscles,three ihnelmuormrbhaargriecgairoenas. STOMACHTrichobezoar Mucosa,erodedin areas SPLEEN Smal Large KIDNEY(S) small Ricgahutd,adlilsyplaced ADRENALRGigLhAtN,Dabsent UTERUS Right hom contained darkbrownfluid vHiosmcsocuosn,tgarieneend-barown substance VPalsaccuelnatraiezastuirornounded bsyubsatthaincck,eyellow OVARIES Parovarian cyst(s) NN % RAONNGE /STUDY % 1 om 01 (1298) 2 02 01 (043) 1011 01 (042) 3100132 0011 ((0024000)) 1011 01 (0200) 101 01 (050) 101 04 (050) 1011 01 (0167) 1011 01 (040) 1011 01 (043) 10M 01 (050) 0 326 05 (0250) 000927 418-012PAGE E-5 `SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS NZW RABBITS P#EORFISOTDUDIES JANUARY 1996 - JANUARY 199685 ##LLIIVTETEFRESTEUXSAEMSIENXEADMINED (DAY 29) 816 6020 ALTERATION SKIN Absentarea RANGE / STUDY No% ON % LF1 1 000112 0011 ((005038)) HEAD Meningocele L 2 026 01 Cyclops F L 3 004 1012 0012 F1 001 01 Upperjawintwo L102 01 Nasregemseanbtssent LF1100012 04 04 Fleshyprotusion ~~ F L 11 000112 0011 F1 001 01 (0467) ((003579)) (007) ((005097)) (053) ((000583)) (0086) EYES Bugedepressed Eyelids open FL 33 000347 0011 ((000583)) Lo 1 012 01 (059) F1 001 01 (007) snout Short FLo 22 00023 0041 ((000579)) TONGUE Protrudes Lo1 012 01 (053) F1 001 04 (008) B80DY Umbilical Homa ~~ L 9 110 02 Edema LF o110 001124 0013 F 1 001 01 (0-500) ((0015230) (008) L: LITTER INCIDENCE F: FETAL INCIDENCE 000928 418-012.PAGE E-6 SUMMARY OF FENTZAWL RGARBOBSISTSEXTERNAL ALTERATIONS ALTERATION N BODY (CSOkNiTn)discolored L 1 purple Spina bifida F3 L2 F2 Hemomhagicareals) LF 2 2 Meningocele FL 22 Hematoma L1 F1 Dark red areas. L1 F1 FORELIMPBaSw(AsN):D/FlOeRxeHd/INDLIMBLS 2 Rotated F2 Paw: Shordigts ~~ L1 Fo1 Limb(syRotated ~~ L F 2 2 Limb(s): Absent L1 F1 TAIL Short L7 F 10 RANGE / STUDY % N 0% 012 01 004 03 024 003 01 04 024 003 041 O1 000234 04 01 012 01 001 04 012 04 001 04 (053) (019) (0200) (019 (050) (006) (0167) (020) (059) (008) (058) (008) 024 01 003 01 012 01 001 01 024 01 003 01 012 01 001 01 (056) (0:08) (056) (007) (0-167) (0-19) (053) (008) 08 014 01 04 (0200) (023) L: LITTER INCIDENCE F: FETAL INCIDENCE 000929 418-012:PAGE E-7 SUMMARY OF FNEZTAWLRSAOBFBTITTISSSUE ALTERATIONS P#ESRTIUODDIES JAN 1996U-JAANUAR RY1Y 9388 ##LFIETTTUESRESSEEXXAAMMIINNEEDD 5677349 ALTERATION BRAIN Diatedlateralventries (Moderate) (Grade2) EYE(S) Circumcomealhemorthage Microphthaimia HEART Large Threeventricies VESSELPSersistenttruncus airntneormiionsaitse, absent Twopulmonary arteries LUNGS Oonrceoomrpmloerteealogbeense,spiarsti~al~ Rilogbhetsafupsediancdcaardilac DIAPHRAHGerMnia N% L 2 030 ~F 2 003 LF21210 20937 L 1 015 F 1 002 LF 11 000125 FL 11 000125 L F 1 1 000125 L 1 015 LF 11 00.0125 F 1 002 LF 6990 1105274 L 1 045 F 1 002 FL 1 015 1 002 RANONGE/S%TUDY 0-1 (05.0) 0-1 (006) 0-2 03 (0-167) (0-19) 01 (050) 01 (0:08) 00441 ((00:5096)) 0011 ((00:5098)) 0014 ((00:5008)) 01 (063) 00441 ((00-0579)) 01 (008) 0095 ((00--2570.)8) 0-1 (053) 04 (0:06) 01 (050) 01 (0:06) FL:i LFIETTTAELRIINNCCIIDDEENNCCEE 000930 418-012:PAGE E-8 SUMMARY OF FETAL SOFT TISSUE ALTERATIONS NZW RABBITS ALTERATION KIDNEY(S) Absent Displacedcaudally SPLEEN Pale GONADS.Righttestsdisplaced caudally RANGE/STUDY N% N % L 1 015 01 (058) F 1 002 04 (008) L 1 015 04 (043) F 1 002 04 (005) L 1 015 04 (07.4) F 1 002 01 (009) L 1 015 01 (059) F 1 002 01 (006) LiLITTER INCIDENCE F: FETAL INCIDENCE 000931 `SUMMARY OF FETAL SKELETAL ALTERATIONS NZW RABBITS 418-012:PAGE E-9 PERIOD JAN19U96-AJARNUAYRY 1998 #STUDIES. 37 #LITTERS EXAMINED 668 #FETUSES EXAMINED 684 #OF STUDIES ALTERATIONS RANGE /STUDY WITH sku N % ON % ALTERATION `Summartzaatlliroenguolafr L185 27.69 0-11 (0667) 35 ossificationofskull F237 am 017 008) Anterior Fontanale Imagularly shaped Lo 1 01s 010083) F102 010005) 1 Posterior Fontanele + E(nGlraardgeed1)(Sight) Lo 1 015 01053) 1 F102 01008 Frontals mogul suture Lo 18 200 o03@t6n 13 Fo 2 035 03018 + ntetrontals present Lu 25 030150 13 Fo 0% 03017 Fused L 5 075 020108 4 F 5 00m 02011 + Two segments L 1 015 01059 1 F100 01007) +Suturelarge. Lo 1 015 01050) 1 Fo1 om 01008) Smal Lo 1 01s 01083) 1 F102 01008 Parieta(s) Contain holes: L 3 045 01058 3 F 3 005 01008 Fusedand small Lo 1 015 01053 1 F102 01008 Inerparietals Lo 1 015 01053) 1 imoguiarly shaped F 1 0m 01005 `inearetals incom L 1 015 010167) 1 pletely ossifed Foo1 0m 01019) L: LITTER INCIDENCE F: FETAL INCIDENCE 000932 418-012PAGE E-10 SUMMARY OFFETAL SKELETAL ALTERATIONS NZW RABBITS #or ALTERATIONS RANGE /STUDY sWTIuTDHiEs SKNaUsLalLs(CONT) N% ON % ALTERATION mogularsure LF4407om 002200420 3 Inomasais LFooox oasmt 004300238 1 Invanasals LFo oote 240 0m 0201) 02019 1 Displaced sure LFootse 322%7 080400) 09052) M Fused LFo0on10 00220011030 9 smal L Fottooo 001100058) 1 NaimsaagluiFarronaanldslourrmessa:lned LE1o3z 0o2w3 0032(0m-19m) 9 Premasiiae: fused LFoooott oo oo 01089) 01009) 1 Promasise: notossfied FL oot1 015 0 041(050) 01008) ' Maxie: used L Fot o1 os oo 01083 01008) ' Ssuhpraapcecdoplals:imoguiaty LF011052 0041(000553)) ' Eye socket: smal LFo z2 0omn 001100s0)8 2 Swhewmossfeston FL 1102015 0011005088 1 Hyon Ata(o,angulated smal Imsguiaty shaped LFowwe 1264117 00188009490)0 om LFoat oooorm 004100284)) 1 LF110021 0011000s8y 1 LF:: UFETTTAELRIINNCCIIDDEENNCCEE 000933 418-012:PAGE E-11 `SUMMOFAFERTAYL SKELETAL ALTERATIONS ZW RABBITS #0F sTupies ALTERATIONS RANGE/STUDY WITH N% ON % ALTERATION VERTEBRAE Cervical iContum, unital L 2 030 010058 2 ossification Fo :AchesandorCena, L 2 0M 01007 3 045 O41 (058) 3 fused +Homivertebra FLo 33 004055 0011005097) 3 F 3 005 01008 iConvum,asymmetric L 1 015 01056) 1 +Conta, bifid Foot om L105 0011008031 1 F102 01008 Thoracic +Hemivertobra Lo 10 150 01089 10 Fotos 0201) ArchesandiorCena, L 5 075 02008 4 fused Conrum, unilateral F600 02042 L 5 075 041053) 5 oisCsiofnivcaa,tioonneormore FLo 15 00105 O04110(00588) 1 asymmetric + Contrum,bifid F102 01008 L 4 o 020108 3 iConva,notossiied Fa om L 1 015 0012004152) 1 +Arch, absent Foot 02 01008 Lt 015 01058 1 +Arch, smal F102 01008 Lo 3 045 010059 3 F 3 005 010008) LumbaHremivertobra Lo 2 0X 01059) 2 Foz oo Centrum, unlatersl ~~ L 1 015 0041005008) 1 o+sAsricfh,castmiaolnl F Lo 12020 01008 01050 2 F200 010008) GContum, notossifed L 1 015 01050) 1 Foot om 010008) Li LITTER INCIDENCE F: FETAL INCIDENCE 000934 418-012:PAGE E-12 `SUMMARY OFFETALSKELETAL ALTERATIONS NZWRABBITS #OF STUDIES RANGE/STUDY WITH ALTERATIONS N % N % ALTERATION VERTEBRAE(CONT) Sacaal +Archesopen Lo 1 ois 01083) 1 Fo 1 002 010008 Caudal :Oneormore misaligned Fused +11 present +12present 131014present 15present +Imeguiarly shaped L 30 440 030167) 24 Fo 31 0s 03(020) Lo 8 120 01(059) 8 F 10 018 03016) L105 01043) 1 Fo 1 002 010008 L105 01043) 1 Fo 2 004 02010) L 3 045 01058 3 F 3 005 01008 L105 01043) 1 F102 010005) Lo 1 015 01080 1 Fo 1 002 01008) VERTEBRAERIS InorolaledVortsbral/R ~~ L 3 045 03 (0158) 1 malformations Fo 3 005 03016) RBS CervicalRibpresent. L 2 0. 01058 2 F 2 004 01(008) Two ormore,fused L 6 0% 020105 5 F 6 010 02042) Basesproximate Lo 10 150 02(091) s F 10 0m. 02011) One ofmore, spit Lo 8 120 010088) 8 F 8 014 01(008) Onoormore, thickenedareas L 27 404 03 (0-167) 19 Fo 2 051 03022 Fiat L 2 030 01088 2 F 2 004 01(006) L: LITTER INCIDENCE F:FETAL INCIDENCE 000935 418-012:PAGE E-13 SUMMARY OF FETALSKELETALALTERATIONS NZW RABBITS #OF STUDIES RANGE /STUDY WITH ALTERATIONS N % N % ALTERATION RIBS (CONT) Extra rib L 1015 01043) 1 F 1002 010008) `Small L 2 030 01(059) 2 F 2 004 01008) Broad L 1015 04 (059) 1 F 1002 O4(008) Bont L 1015 04 (045) 1 F 1002 01(008) MANUBRIUM Duplicated Fused L 1015 01059) 1 F 1 002 01007) L 3 045 02 (0-105) 2 F 3 005 02(01) STERNEBRAE Twoormore,fused L 84 958 05028) 31 Fo 7 137 07039) One ormore,asymmetric L 6 080 02 (0105) 5 F 6 010 02(042) One ormors, incompletely L 11 165 02 (0-105) 9 ornotossified Foot 019 02012) Duplicated L 1015 041059) 1 F 1002 04007) PELVIS Pubis(es): incompletely L 4 080 01059) 4 or not ossified F 5 000 02011) SCAPULAE Aa(e): imeguiarly shaped Aa(e):wavy L 4 080 02 (0108) 3 F 4 007 02(012) L 1015 04 (053) 1 F 1002 01 (008) L: LITTER INCIDENCE F: FETAL INCIDENCE 000936 418-012:PAGE E-14 `SUMMARYOFFETALSKELETAL ALTERATIONS NZWRABBITS #OF STUDIES. RANGE/STUDY WITH ALTERATIONS N % ON % ALTERATION `SCAPULAE (CONT.) Misaligned Bent L 5 om o2@08 4 F 5 0m 02012) Lo 1 015 010080) 1 F110: 01008) FORELIMB(S) 1 Phalan present Lo 1 015 01088) 1 Fo 1 0m 01007) 0Phalanges present. Lo 1 015 01088) 1 Humerus,Radius, Uina, Fo 1 0m 01007) Carpals, Metacarpals,Fore- digits andForephalanges L 1 015 04 (0-53) 1 absent Foot 002 04(008) L: LITTER INCIDENCE F: FETAL INCIDENCE 000937 APPENDIX F STATEMENT OF THE STUDY DIRECTOR 000938 %PRIMEDICA 418-012:PAGE F-1 Sahoipteae fe PROTOCOL 418-012: ORAL (STOMACH TUBE) DEVELOPMENTAL TOXICITY STUDY OF PFOS IN RABBITS `SPONSOR'S STUDY NUMBER: 6295.10 STATEMENT OF THE STUDY DIRECTOR This final report accurately reflects the raw data obtained during the performance of the study. No significant 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 on Drugs 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 ofgoodlaboratory practice' occurred that affected the quality or integrity of the study. - wt oe 10-00-99 Frond G. Yor(Pr ,DABT Date AS? Directokof-Research 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 (1988). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32(No. L 315; 28 October): 1-17. 000939 APPENDIX G `QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT 000940 r2, PRIMEDICA 418-012:PAGE G-1 ArgusrRaphensrnhshy:LBaDboa Ghrat1osrai9es,B2ai43l2ig7In1nc?0. 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 418-012: Oral (Stomach Tube) Developmental Toxicity Study of PFOS in Rabbits Sponsor's Study Number: 6295.10 "The draft protocol for this study was audited for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles. of good laboratory practice on 10 AUG 98. Critical phases of this study were inspected five times; study information and raw data were audited twice (see tables 1 and 2 for dates and phases/data). The draft final report and the raw data for this study [except for Appendix F, the Pilot Report, which was conducted in the spirit of Good Laboratory Practice (GLP)] were compared and audited for accuracy, for adherence to protocol requirements, and for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice between 04 DEC 98 and 17 DEC 98, for revisions requested by the Sponsor 23 DEC 98, and for finalization on 11 JAN 99. 000941 418-012: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 Drugs, and 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 laboratory practice. soi ametesi. Barbara J. terson, BA. Directorof Operations and Compliance Date Hu|.tflbih tineuungennts Heather L. Rabuttino, M.S. Date Quality Assurance Supervisor and Principal Auditor 000942 TABLE 1 CRITICAL PHASES INSPECTED 418-012.PAGE G-3 Test Article Administration - Gavage Date of inspection: 25 AUG 98 Date results reported to the Study Director and Management: 12 SEP 98 Test Article Preparation Date of inspection: 04 SEP 98 Date results reported to the Study Director and Management: 12 SEP 98 Blood Collection Date of inspection: 10 SEP 98 Date results reported to the Study Director and Management: 10 SEP 98 Caesarean-Sectioning Dates of inspection: 10 SEP 98, 18 SEP 98 Dates results reported to the Study Director and Management 10 SEP 98, 24 SEP 98 000943 TABLE 2 418-012PAGE G4 RAW DATA AUDIT(S) on 1T2heOCfoTll9o6w:ing study information and raw data were audited TVeehsitcalretircelceeriepcte,ipptr,epparreaptairoantiaonndaunsde.use. Test article packing lists. ManTahgeemreesnutltosnof13thiOsCaTudi9t6.were reported to the Study Director and on 1T3heOCfoTll9o8w:ing study information and raw data were audited Protocol. PLirsottoofcoplerasmoennndemleanntds.computer operator codes. Error codes and codes for clinical sign observations. `Animal receipt, randomization, and acclimation. Veterinary examination. FIne-leidfe ctornasnsuamcpttiioonnr.ecord. Caesarean-sectioning. FMeattaelrngarlosgsroosbsseorbvsaetrivoantsi.ons. Fetal fixative assignment. Fetal visceral examination. Fetal skeletal examination. NOercgraonpswye.ights. Tissue packing lists. `General comments. TSetmupdsycrmiabienst.enance records. Feed and water analyses EDdoistargeequvesotlsu.mes. Deviations. BDlaotoadrceovlileewctpiaogne. 000944 The resis of this audit were reported to the Study Director and Management on 14 OCT 98.