Document qkb8q2d809Y8O0BG08o324qxE

FINAL REPORT RECEIVED QFPT CBIC 200FER22 AHiL3Y PROTOCOL 418-008 COMBINED ORAL(GAVARGEPER)OFDEURTCITLIITOYN. TDOEXVICEILTOYPSMTEUNDTYALOFANPDFOS PERINATAUPOSTNATAL IN RATS SPONSOR'S STUDY NUMBER: 6295.9 FINAL REPORT DATE: 10 JUNE 1999 000502 PROTOCOL 418-008 PERCIONMABTIANUEPDOOSRTANLAT(AGLAVRAEGPER)OFDEURCTITLIIOTYN,TDOEXIVCEILTOYPSMTEUNDTYALOFANPDFOS INRATS SPONSOR'S STUDY NUMBER: 6295.9 TABLE OF CONTENTS SUBJECT PAGE I. SUMMARY AND CONCLUSIONS - A. Methods 1 B. Results 5 C. Conclusion 10 Il. DESCRIPTION OF TEST PROCEDURES I1 A. Conduct of Study 1-1 A. Sponsor 1-1 A2. Testing Facility 11 A3. Study Number 1-1 A4. Sponsor's Study Number 1-1 AS. Purpose of the Study I AS. Study Design 11 AT. Regulatory Compliance 2 ASB. Ownership of the Study 2 AS. Study Monitor 2 i 000503 SUBJECT A10. Alternate Study Monitor A.11. Study Director A.12. Technical Performance A13. Report Preparation A.14. Report Review A.15. Date Protocol Signed A.16. Datesof Technical Performance A17. Records Maintained B. Test Article Information B.1. Description B2. Lot Number B.3. Date Received and Storage Conditions B.4. Special Handling Instructions B5. Analysis of Activity C. Vehicle Information C1. Description C2. Lot Numbers C.3. Dates Received and Storage Conditions C4. Special Handing Instructions C5. Analysis of Purity D. Test Atticle Preparation D1. Sample Information ii 000504 PAGE 12 2 2 1-2 1-3 3 3 4 4 14 4 15 5 Is 5 Is Is 15 16 [1 6 16 SUBJECT D.2. Analytical Results E. TestSystem Ed. Species E2. Strain E3. Supplier (Source) E4. Sex ES. Rationale for Test System ES. Test System Data E.7. Method of Randomization EB. System of Identification F. Husbandry F.1. Research Facilty Registration F2. Study Rooms F.3. Housing Fa. Lighting FS. Sanitization F6. Feed F7. Feed Analysis F8. Water F.9. Water Analysis F.10. Bedding F.11. Bedding Analysis il PAGE 7 7 7 7 7 7 7 7 18 I8 1-9 Ig Ig Ig 1-10 I-10 I-10 110 110 I-10 I-11 I-11 000505 SUBJECT G. Methods G.1. Dosage Administration G2. Assigned Rat Numbers G3. Rationale for Dosage Selection G4. Route of Administration G5. Rationale for Route of Administration G6. Frequencyof Administration G.7. Length of Study G8. Method of Study Performance G.9. Gross Necropsy G.10. Statistical Analyses Ill. RESULTS - Fo GENERATION MALE RATS A. Clinical Observations 8B. Body Weights and Body Weight Changes C. Absolute Values (giday) and Relative (g/kg/day) Feed Consumption D. Mating and Fertiity E. Necropsy Observations F. OTrergmainnaWleiBgohdtytWoeTiegrhmtisn,alOrBgoadnyWWeeiigghhtts and Ratios (%) of IV. RESULTS - Fo GENERATION FEMALE RATS A. Clinical Observations Al. Mortality PAGE I-11 1-11 I-12 I-12 I-12 112 I-12 1-13 1-13 I-18 I-21 1 1 1 1-1 n-2 n-2 0-3 v-1 v-1 v1 iv 000506 susJECT A2. Clinical Observations B. Body Weights and Body Weight Changes B.1. Precohabitation B.2. Gestation B.3. Lactation C. VaAblsuoelsute (g/day) and Relative (g/kg/day) Feed Consumption C.1. Precohabitation C2. Gestation C3. Lactation D. Estrous Cycling, Mating and Fertility E. Necropsy Observations F. Caesarean-Sectioning and Litter Observations G. Natural Delivery and LitterObservations H. NCleicnircoaplsOybOsbesrevravtaitoinosnfsrom Birth to Day 21 Postpartum and I Reflex and PhysicalDevelopment 1, Surface Righting 2 Pinna Unfolding 13. Eye Opening 14. Acoustic Startle 15. Air Righting 16. Pupil Constriction PAGE v1 v1 v-1 v-2 v2 v3 v3 v-3 v-3 v4 v4 v4 v4 V5 v-6 v-6 v7 v7 v7 [24 v8 v 000507 SUBJECT V. RESULTS - F1 GENERATION MALE AND FEMALE RATS A. F1 Generation Male Rats A. Mortality and Clinical Observations A2. Body Weights and Body Weight Changes A3. AVbaslouleuste (g/day) and Relative (g/kg/day) FeedConsumption Ad. Sexual Maturation AS. Passive Avoidance Performance AS. Watermaze Performance AT. Mating and Fertility AB. Necropsy Observations A. Terminal of Organ WBeoidgyhtWetiogThetrsmiannadl Organ Body Weights Weight and Ratios (%) B. F1 Generation Female Rats B.1. Mortality and Clinical Observations B.2. Matemal Body Weights and Body Weight Changes B3. CMoantseummapltAibosnolVuatleue(sg/day) and Relative (g/kgiday) Feed B.4. Sexual Maturation BS. Passive Avoidance Performance B6. Watermaze Performance B.7. Mating and Fertilty B.8. Necropsy Observations B.9. Natural Delivery and Litter Observations vi 000508 PAGE \Z] V-1 \Z] v2 v2 va v3 v3 v-3 v3 v4 v4 v4 v5 v6 v7 v7 v7 v7 v8 v8 SUBJECT 8.10. CNleicnircoaplsOybsOebrsvearvtaitoinosnfsrom Birth to Day 21 Postpartum and REFERENCES APPENDIX A - REPORT FIGURES PAGE V-9 V-10 Figure 1. Body Weights ~ Fo Generation Male Rats A Figure 2. Body Weights ~ Fo Generation Female Rats A2 Figure 3. Body Weights ~ F1 Generation Male Rats A3 Figure 4. Body Weights ~ F1 Generation Female Rats Ad APPENDIXB - REPORT TABLES - Fo GENERATION MALE RATS Table B1. Clinical Observations - Summary -- Fo Generation Male Rats B.1 Table B2. Body Weights ~ Summary - Fo Generation Male Rats B2 Table B3. Body Weight Changes ~ Summary -- Fo Generation Male Rats 8.3 Table B4. AFbosoGleunteeraFteieond MCaolnesuRmapttsion Values (g/day) -- Summary B4 Table BS Relative Feed Fo Generation Consumption Male Rats Values (g/kg/day) ~ Summary -- B-5 Table 86. Mating and Fertity - Summary - Fo Generation Male Rats B.6 Table B7. Necropsy Observations ~ Summary - Fo Generation Male Rats B.7 Table BS. TFeormGiennaelraBtoidony MWaelieghRtastsand Organ Weights ~ Summary -- 88 Table BS. SRautmimosar(y%)-oFfoOrGgeannerWaetiigonhtMtaolTeeRramtisnal Body Weight -- 8.9 Table B10. CMlailneicaRlatOsbservations - Individual Data ~ Fo Generation 8-10 Table B11. Body Weights - Individual Data ~ Fo Generation Male Rats B.20 vit 000509 SUBJECT PAGE Table B12. FFoeeGdenCeornastuimopntMiaolneVRaaltuses - Individual Data - B-30 Table B13. MMaatlienRgaatnsd Fertility ~ Individual Data -- Fo Generation B40 Table B14. MNaelceroRpastysObservations ~ Individual Data ~ Fo Generation 8-50 Table B15. Terminal of Organ Body Weights and Weight to Terminal Organ Body Weights Weight ~ and Ratios Individual (%) Data - Fo Generation Male Rats B58 APPENDIX C - REPORT TABLES ~ Fo GENERATION FEMALE RATS. Table C1. RaCltinsical Observations ~ Summary -- Fo Generation Female C1 Table C2. FBeomdaylWeeiRgathsts Precohabitation ~ Summary Fo Generation c4 Table C3. FBoodGyenWeeriagthitonChFaenmgaelse -RaPtrsecohabitation ~ Summary - cs Table C4. MFaotGeernnearlaBtoiodny WFeeimgahltesR~atGsestation ~ Summary -- [= Table C5. Maternal Body Fo Generation Weight Female Changes Rats -- Gestation ~ Summary -- cs Table C6. FMaoteGrennaelraBtoiodny WFeeimgahltesR~atLsactation ~ Summary -- co Table C7. FMaotGeernnaelraBtoidony FWeemigahlte CRhatasnges -- Lactation ~ Summary c-10 Table C8. APrbescoolhuatbeitFaeteidonC~onSsuummmptairoyn~VaFlouGeesn(egra/tdiaoyn--)Female Rats C-11 Table C9. PRreelcaotihvaebiFteateidonCo~nsSuummpmtairoyn Values (g/kg/day) -- - Fo Generation Female Rats C12 Table C10. GMeastteartniaolnA~bsSoulumtmearFeye-d FCoonGseunmeprtatiioonnVFaelmueasle(gR/adtasy) ~ c13 vii 000510 SUBJECT Table C11. GMeastteartniaolnR~elSatuimvmeaFreyed~ CFoonGseunmeprtaitoinonVaFleumeasl(eg/Rkagt/sday) Table C12. LMaactteartniaoln A--bsSoulumtmeaFreey~dFCooGnesnuemrpattiioonn VFaelmuaelse(Rg/adtasy) - Table C13. MLaactteartniaoln R--elSautmivmeaFreye~d FCoonGseunmeprtaitoinonVFaleumeasle(gR/aktgs/day) -- Table C14. Table C15. FEostGreonuesrCaytciloinng,FeMmaatliengRaatnsd Fertility -- Summary -- FNeemcraolpesRyaOtbsservations ~ Summary ~ Fo Generation Table C16. FCaoeGseanreeraant-iSoenctFieomnailnge Observations Rats ~ Summary -- Table C17. Natural Delivery Observations -- Summary Fo Generation Female Rats Table C18. Litter Observations (Naturally F1 Generation Litters Delivered Pups) -- Summary ~ Table C19. Table C20. CSluimnimcaalrOyb-seFr1vaGteinoenrsaftrioomn BPiurtphsto Day 21 Postpartum -- Reflex and Physical Development -- Summary -- F1 Generation Litters Table C21 Table C22. Necropsy Observations -- Summary -- F1 Generation Pups Clinical Observations ~ Individual Data -- Fo Generation Female Rats Table C23, Table C24. Table C25. FBoodGyenWeeriagthitosn --FePmreacloehaRbaittsation -- Individual Data -- Maternal Body Weights -- Presumed Gestation -- Individual Data ~ Fo Generation Female Rats Maternal Body Weights -- Lactation -- Individual Data -- Fo Generation -- Fo Generation Female Rats PAGE C14 C15 C16 C-17 cg c20 c-21 c-22 c-25 C-26 Cc-31 C-32 c40 C-50 C-70 ix 000511 SUBJECT PAGE Table C26. DFaetead-CoFnosGuemnpetriaotnioVnalRuaetss -- Precohabitation Individual crs Table C27. Maternal Individual Feed Data C--oFnosuGmepnteiroantiVoanlFueesmalPerReastusmed Gestation -- ces Table C28. IMnadtievrindaulalFDeaetda C--oFnosuGmepnteiroantiVoanlFueesma--lLeacRtaattsion c-108 Table C26. FEsotGroeunserCaytciloinngFeamndalDeayRastisn Cohabitation -- Individual Data -- c-110 Table C30. FNeecmraolpesRyaOtbsservations -- Individual Data -- Fo Generation C115 Table C31. FCaoeGseanreeraant-iSoenctFieomnailnge Observations Rats -- Individual Data c-121 Table C32. LiItndtievriOdbusaelrDvaattiao~nsF1(GCeaneesraartein-oDneLliitvteerrsed Embryos) -- C126 Table C33. LiEtmtberrysonal Vital Status ~ Individual Data ~ F1 Generation C131 Table C34. SNaetxurIanldiDveliidvuearyl,DIamtpala~ntFaotiGoennSeirtaets,ioanndFePmuapleViaRbaitlsi/ty and F1 Generation Litters C136 Table C35. PPousptpBaordtyumWe~igInhdtivLiidttuearlADvaetraag--eFs1fGreonmerBiarttihotno LDiattyer2s1 c-141 Table C36. Pup Body Individual DWaetiagh-tsF1frGoemneBriratthioton Day 21 Pups Postpartum C146 Table C37. Pup Vital Individual Status and Data ~ F1 GSeenxerfartoimonBiPrtuhptso Day 21 Postpartum -- C176 Table C38. ~ ICnldiniivciadluaOlbsDeartvaa~tiFo1nsGefnreormaBtiirothn tPouDpasy 21 Postpartum -- c-181 Table C39. Surface Righting - Individual Data -- F1 Generation Litters C-182 Table C40. Pinna Folding ~ Individual Data - F1 Generation Liters C-193 * 000512 SUBJECT Table C41. Eye Opening - Individual Data -- F1 Generation Liters Table C42. Acoustic Startle ~ Individual Data - F1 Generation Litters Table C43. Air Righting ~ Individual Data ~ F1 Generation Litters Table C44. Pupil Constriction Individual Data ~ F1 Generation Litters Table C45. Necropsy Pups Observations ~ Individual Data -- F1 Generation APPENDIX D - REPORT TABLES ~ F1 GENERATION MALE RATS Table D1. Clinical Rats. Observations ~ Summary - F1 Generation Male Table D2. Table D3. Body Weights -- Summary -- F1 Generation Male Rats Body Male Weight Rats Changes -- Summary -- F1 Generation Table D4. Absolute Feed F1 Generation Consumption Male Rats Values (g/day) ~ Summary -- Table D5. Relative Feed F1 Generation Consumption Male Rats Values (g/kg/day) ~ Summary Table D6. Table D7. Sexual Maturation ~ Summary - F1 Generation Male Rats PF1asGseinveerAavtoiiondaMnacleePReartfsormance ~ Summary -- Table D8. Watermaze Male Rats Performance -- Summary -- F1 Generation Table D9. Mating and Fertlty ~ Summary - F1 Generation Male Rats Table D10. Necropsy Observations Male Rats -- Summary -- F1 Generation Table D114. Terminal Body F1 Generation Weights and Male Rats Organ Weights -- Summary -- PAGE -203 C.213 C223 C-233 C243 D-1 D3 D4 D5 D6 D.7 Dg Dg D-10 D-11 D-13 xi 000513 SUBJECT PAGE Table D12. RSautmimosar(y%)-oFf1OrGgeannerWaetiigohntMtaoleTeRramtisnal Body Weight -- D-14 Table D13. MClailneicaRlatOsbservations ~ Individual Data -- F1 Generation D-15 Table D14. Body Weights ~ Individual Data ~ F1 Generation Male Rats D-20 Table D15. MFaeleed RCaotnssumption Values ~ Individual Data ~ F1 Generation 0-26 Table D16. RSaetxsual Maturation - Individual Data ~ F1 Generation Male D-29 Table D17. FP1asGseinveerAavtoiiondaMnacleePReartfsormance -- Individual Data - D-30 Table D18. WMaalteerRmaatsze Performance - Individual Data -- F1 Generation 0-33 Table D189. RaMattsing and Fertility ~ Individual Data -- F1 Generation Male D3 Table D20. MNaelceroRpastysObservations -- Individual Data -- F1 Generation D-39 Table D21. Terminal of Organ WBeoidgyhtWetiogThetrsmiannadl Organ Body Weights Weight -- and Ratios (%) Individual Data -- F1 Generation Male Rats D43 APPENDIX E - REPORT TABLES~ F1 GENERATION FEMALE RATS Table E1. RaCltinsical Observations ~ Summary -- F1 Generation Female E1 Table E2. Body Weights Female Rats ~ Precohabitation -- Summary ~ F1 Generation E4 Table E3. BF1odGyenWeeriagthitonChFaenmgaelse --RaPtrsecohabitation - Summary ES Table E4. F1MaGteenmealraBtoiodny FWeeimgahltesR~atGsestation -- Summary -- E6 xi 000514 SUBJECT Table ES. Materal Body Weight Changes - Gestation -- Summary F1 Generation Female Rats PAGE E-8 Table E6. Table 7. Table ES. MF1atGeernnearlaBtoiodny WFeeimgahltesR--atLsactation -- Summary -- Matemal Body Weight Changes - Lactation -- Summary -- F1 Generation Female Rats Absolute Feed Consumption Values(g/day) -- Precohabitation -- Summary ~ F1 Generation Female Rats E-9 E-10 E-11 Table ES. Relative Feed Consumption Values (g/kg/day) -- Precohabitation -- `Summary -- F1 Generation Female Rats E-12 Table E10. Maternal Absolute Feed Consumption Values (g/day) -- Gestation -- Summary ~ F1 Generation Female Rats E-13 Table E11. Maternal Relative Feed Consumption Values (g/kg/day) -- Gestation -- Summary - F1 Generation Female Rats E-14 Table E12. MLaactteartniaoln A--bsSoulmumtearFyee-dFC1oGnesnuemrpattiioonn VFaelmuaelse(Rg/adtasy) ~ E15 Table E13. Table 14. LMaactteartniaoln R-elFa1tiGveeneFreaetdioCnonFseummaplteioRnatVsalues (g/kg/day) -- E16 Sexual Maturation -- Summary - F1 Generation Female Rats E-17 Table E15. Passive Avoidance Performance -- Summary -- F1 Generation Female Rats E-18 Table E16. WFeamtaelremaRzaetsPerformance -- Summary - F1 Generation E-19 Table E17. Mating and Fertility -- Summary ~ F1 Generation Female Rats Table E18. FNeemcraolpesRyaOtbsservations ~ Summary -- F1 Generation E-20 E21 Table E19. FNaetmuarlael DRealtisvery Observations -- Summar-y F1 Generation E-22 xii 000515 SUBJECT Table E20. Litter Observations (Naturally Delivered Pups) -- PAGE Summary - F2 Generation Litters E23 Table E21. SCluimnimcaalrOybs-eFr2vaGteinoenrsaftrioomn BPiurpthsto Day 21 Postpartum E26 Table E22. Necropsy Observations ~ Summary F2 Generation Pups E27 Table 23. FCleimnaiclael ORbastesrvations ~ Individual Data ~ F1 Generation E28 Table 24. GBeondeyraWteiiognhtFsem-aPlreecRoahtasbitation ~ Individual Data -- F1 E34 Table E25. Maternal Data ~ F1 BGoednyerWaetiigohntsFe~mParleesRuamtesd Gestation -- Individual E37 Table E26. FM1atGeernnearlaBtoiodny FWeeimgahltesR--atLsactation -- Individual Data -- E43 Table E27. DFaetead~CoFn1sGuemnpetriaotnioVnalFueemsal--ePrReactoshabitation ~ Individual E45 Table 28. Maternal Individual Feed Data C-oFn1suGmepnteiroantiVoanluFeesma~lPerReastusmed Gestation -- E49 Table E29. DMaattear~naFl1FGeeenderCaotnisounmFpetmioanleVaRlautess ~ Lactation -- Individual E52 Table E30. FSeexmuaalleMRaattusration ~ Individual Data - F1 Generation E55 Table E31. FP1asGseinveerAavtoiiondaFnecmeaPleerRfaotrsmance -- Individual Data -- E56 Table 32. FWeamtaelremaRzaetsPerformance - Individual Data - 1 Generation E59 Table E33. FDeamyaslien RCoahtasbitation - Individual Data - F1 Generation E62 Table E34. FNeemcraolpesRyaOtbsservations ~ Individual Data -- F1 Generation E63 xiv 000516 SUBJECT Table E35. FNSa2etxuGre-anleInrDdaeitlviiivodenurayLl,itDItaemtrpsala--ntFa1tiGoennSeirtaetsi,oannFdePmuapleViRaabitlsi/ty and PAGE E-67 Table E36. PPousptpBaordtyumWe--igInhdtivLiidtutearl ADvaetraa--geFs2fGreonmeBriarttihotno LDiattyer2s1 E-70 Table E37. IPnudpiviBdoudaly DWaetiagh~tsF2frGoemneBriartthioton DPauyps21 Postpartum -- E73 Table E38. Pup Vital Status and Sex from Birth to Individual Data -- F2 GenerationPups Day 21 Postpartum -- Table 39. Clinical Observations from Birth to Day 21 Postpartum -- E-91 Individual Data -- F2 GenerationPups E-94 Table E40. Necropsy Observations Pups Individual Data ~ F2 Generation E-95 APPENDX F- PROTOCOL AND AMENDMENTS F-1t0 F-63 APPENDIX G - DEVIATIONS FROM THE PROTOCOL ANDTHE TSETSATNIDNAGRFDACOIPLEIRTYATING PROCEDURES OF THE G1 APPENDIX H - RTEEPMOPRETRSATURE AND RELATIVE HUMIDITY HA to Ho APPENDIX |- STATEMENT OF THE STUDY DIRECTOR 1 APPENDIX J - SQUTAALTIETMYEANSTSURANCE UNIT FINAL REPORT J-1t0J-8 xv 000517 TITLE: 418-008:PAGE I-1 ASCTNOUDMDBPYIENOREIFDNPAOFTROAASLL/IP(NOGRSAATVTNASAGTEA)LFERRETPILRIOTDYU,CDTEIVOENLOTOPXMIECNITTAYL APRRGOTUOSCROELSNEUAMRBCEHR:LAB41O8R-A0T0O8RIES, INC. SPONSOR'S STUDY NUMBER: 6295.9 I. SUMMARY AND CONCLUSION A. Methods Text Figure 1 provides a schematic of the study design. A. Fo Generation Rats/F1 Generation Litters: pDOeanrwelseehyxu)npdrearrtesddowsseeravegeenatsygs-rfioiugvpne.edmTathloeefiarvanetdsdofwseeamrageleaedgCmriio:nuiCpssDte(rGBrRouVpsAF|/tPhlruosugh(VS)p.ra3g8urea.ts the vehicle, 0 (vehicle), 00..15, %0.T4,we1e.6na8n0d,3.o2ramlgl/yk(gv/idaagy.avaTghe)e.maTlheeedrdtahotessatwegesertsearwdteiocrsleee,dPoFnOcSe. or gpbaeeifflooyrreebecsgoaichniainbfiiintcgae.t4i2oTnhdeaanyfdsemcbaoelnfteoirrneauticsnogwhaetbrhiertoadutoigoshneDdaGnodnc9ceo(nrdtaaitisnluyaisbnsgegtihnrnoiunggh4t2hedadyasy 7SecDtiLo*ni2n0g)(.raDtsGth2a4t (driadtsdealsisviergnaedlitteor)n.atTuhrael delivery that diidgnnoetd dosage volume was to Cagagreandeliver a Iter), 5mUkg. dASltolusdFayogaegnepdneerfirooardt.ciloinBniocrdaaltysswwiegeinrgsehotofsbesafefnredcvtefsedeoffdotrchvoeinatsbeusimtlpyatraittioclleeatswtitcweidcaeildyaidluyrdiunrgintghethe ffeemcaorldeerdatwsewekelrye druercionrgdetdhewdeoesklaygetopceorhiaobditaantdioant,sdaacnirilvfyaicldeuu.reisnBfgoordthmyeawlgeeeisgtrhaatttssiowfnoerre dspauecrrriiionfdgi,cteo.hne FgDeeLseStdat1c,ioo4nn,spu7,emrp1it0oidaoanndnvda1lo4une(rsDaLtwssear1se,s4ri,egcn7o,erd1d0teodannwadeteurkally dteolicvoehrayb)itaantdioant, daily natural delivery) 14 (rats assigned to # aDreetapirloevdiddeesdcriin ptthieonaspporfoaplrl ipartoecseedcutrieosnsusofedthiins (PROTOCOL AND AMENDMENTS). trheepocrtonadnudctinofAPthPisENstDuIdXy F b. DDlGiissuusseeddaass aann aabbbbrreevviiaattiioonn ffoorrddaayy ooffl(apctraetsiuonmeodr)dgaeystpaotsitopnartum, 000518 418-008:PAGE I-2 paTeshrsemiigftintresetddttetoon Cnfaaetemusaralarleelayrandt-esslepivceetrridolointsitenarsgg. eonTghrDeoGuspe1w0ri.atthsTawheecrorenefmiarimneidngdafteemaolfemartaitsngwewreere aQtbsbeirrtvha.tiMoantserdnuarlinbgehpaarvtiuorirtioofn,thdeurdaatmiosnwoafsgeesvtaaltiuoant,edelivdtaaelirulsayitwzeehdeafnnodrtchpleiunppivcuiaaplbsilwietrye examined during the 21-day postpartum period. iPEnoastchthepalpriuttepursmwwpaeesrrieeovdra.elcuPoaurtdpeesddfionornecvaiecabhdialliiitltyyearftowrlee2ra1estdctaowyuisnctepeoedsatocnhcdeadyaidlyu.rinPghytshieca2l1-sdiagyns aSanunrddfaaoicbresreirrgihgvtheitdnignngruerrfeslfilenexgx.wbpeihnanvaiournfwoeldriengr,eceoyredeodpeonninDgL,sapca1or(utbsuitmrit.hc),SPt4au,rpt7l,be o1rd4eysapwnoednicg3eh1ts walal spuepvsaliunattheed loitntcereroenaceDhrLeed2m1to.hneiOtcornirteDedrLido4un,rfiaonrgttathbhelees2poef1c-ridfaaincydtpoeosmts.tunpPiatursptiwulamcsopneusrstireeiddcttuioonni locifutltlreralfintrdtoeormsmGtuornofiutosuprswmaIa,slIeuasanendddItlfolofsuoerrlfececotmnattliwenoupemudaplsee,vaawlunhadetritoewn,opofsesimballee. pOupnsDfLro2m1b.a8cthable. PpFreoorsigtoeadnteearnaadtnidnoenscermmoaiplnseiaelrdav;tessgiwrceolsreses l(sewasicitrhoinfaiscnwedderwaieftterhreotucationmfelpduli.ed)tiTwoheneotfestthees,coehpaibdiitdaytmiiodnes, weighed and retained re excised, individually l6Feosigo1en0nseawrnaedtrienoenrcefrtaoeipmnsaeilde.ed;raTpthsreeagsrnastaisngnwceeydrsettaoetCxuasaemwsiaanrseedaconfn-ofsriertcmhteeido.nnuimOnbvgawreirese asancdrigfriocseds on of corpora embryos. Elmutberayionsewaechreovdairsycaarnddedaifmptlearnteaxtaimoinnastiitoesn,. and viaebrleaanndddinsotnivbiuatbiioen aFnodgenneecrraotpisoiendf.emOavlaerireastsaansdsiggrnoesds distribution of implantation sites was tloesniaotnusrawledreelirveetrayinweedr.e recorded sacrificed on DL The number and 21 TsAailbrsesdcahllelitdoeuwrelsdeodtfosdanacarmtiusfriacalelllyaofdtweeelrdicvtoeormpnaalteiuttreiarol)nl,yobdfleotlhoievdecsroaahmalpbilitttear)tiaonndpoenrioDdL(2m1al(efermaatslethat praetr) dwoesraegceolglreoctuepdafnrdomshtihpepiendfetroiotrhveeSnpaocnasvoar fforromphfeaisrvem(amacapolpkerionaxenitmdiactfieavlneyafl4yesmmiaLsl.,epearts pnPeuucpprssopfsnriooetmds.feilveTechotefedtshtfeoormlcaarocgnhetsictnoutneetdernsetvsainl(umGairtlokiuocnpusrodnI). DwIlLearn4ewceorlleecstaecrdiffircoemd aallndculled the Sponsor for analysis d Ii frozen and shipped to 000519 418-008:PAGE I-3 The was flirvoezrefnfoamndeaschhiprpaetdwtaostehxecSispeodn,swoerifgohreadn,alaynsids.a the ters ofthe five dams in Groups | through IV sTahmepllievesresoctfiofnhe(lpauteprsal lobe) from Ssapmopnlseorcoflorleacntailoynsiwse.reDeaxmcisseidn,thpeoo3l.e2dmpge/rkgt/edra,ysfdreoolszeeacntgeeadngfdrorosuphphiap(prGemrdaocutaopktVih)meedtidi not have surviving pups on DL 21. A2. F1Generation Rats/F2 Generation Litters: FgO1ennlegyreanttheieroa0nt.i1boenacnpaduup0ss.e.4 omfgt/hkege/xdcaeyssdiovseagtoexigcritoyupsseewnerientchoent1i.n6uaenddin3t.o2 tmhge/skegc/odnady a(TnGhrdeorfueepmwsael|retehrr1ao5tus0gwhmeaIrl)ee,ga2in5vdernaftaespmpaprelorepsrFei1axtgpeeendreordsaoatsigaoegnsoerfagttrsohuiepn.theF1thgreeneedraotsiaognergeraoisups (gavage) beginning on DL 22 and continuing through thteedstayarbteifcoroeraslalcyrifice. weBeaercgiehnednvoianslgaugaaette2gd4rofdouarpytswheeorfaegaegteeo,sftoevdnaegiinmnaaallpeapsarstaietvnaecnyadvbooeingdienafnecmealpaerraadtifgrmo.m eFaecmhalieterratisn pwoesrtepaevratluumadteady f7o0r,tohneeagmealoef rpartepauntdiaolnseepfaermaatlieonrabtengfiirnnognmioenngaDcohLnl2iD8tt.Ler3Mw4.aelreeOnrate evaluated in a dosage group wweatreer-afislsliegdnMe-dmtaozceo.habOintataipopnr.oximately DL 90, rats within each 4pFe!1riFgo1edngeaernnadetrinaoetnciromonaplsfeieemrdaa,ltseawsreaptrrseewvseiaorcuresilfpiyecrdemedistacftrteiedrbetcodomnfapotrluetrthaileloFynoodfgetnheercaothiaobnitmaatlieonrats. dtheastcrdiebleivdefroerdtlhietteFros wgeerneersaatciroinfifceemdaolne DL 21 rats. and necropsieeldi,vearspfirtteervsi. ouAslllydams FlOe1nsigoBenLnse.2r1aN,teiacollrnoFpp2uspgyse.pnreorcateidounrepsupwserweertheessacarmifeicaesd tahnodseexdaemscirniebdedfofrogrrtohses 000520 xi B. Results 418-008:PAGE I-5 B.1. FGo eneraMtaleiRoatns: tPNhFoeOtFSeo.stgeAalrnlteircclalietniiaconanldmonabolsteersvriaagttnssioodnfisecd(odomtuhpreiornugtnhtdahintsonxsiotcriutdmyy.a)aswearreescuolntsoifdterreeadtmuennrtelwaittehd to Groups reduced abdomdinyiwsetiegrhetd g0a.i4nmsgf/okrgt/hdeaeyntainrde higher study. dosages of th test article had cA3ob.h2saombligutltaektigaodnnadpyerrdeilooastdi,avgeaebfsgeoerldouutcpeosnfsefouerdmtpchtoeinoesnnutimvrapeltuiperosencwovehaarlbeuietrsaetwdieuorcnedperiinotd.he A1.6ftearntdhe reduced in the 0.4 and 1.6 mg/kg/day dosage groups. e significantly aDnodsafegretislitoyfptahreatmeesttearrstieclvealausatheidg.h as 3.2 mg/kg/day did not affect anymating hasdiganliifgihctanbtlryoiwnncroerabsreodwnnulmibveer,raonf mobasleervraattsioinn tahtteri3b.u2temdg/tokge/fdfeacytsdoofsaPgEegOr.oup pbTrohodesyt1aw.te6eigawhnetdrs.e3.s2Tighmneigfkaicgbasinodtllauyytredewdoeusicageghdetsignrotofhutephs3e.hs2aemdmgi/snikaggln/ivdfeiscaincdtlleys rweitdhucfeludidtearnmdintahle ay osage group. B.2. Fo Generation Female Rats/F1 Generation Litters: No Fo PFOS. generation female rats died during this study as a result of treatment with gOebssteartviaotniaonnsd olfacltoactailoinzpederailoodpseciniathweer0.e4,in1c.r6eaansedd3d.u2rmigng/ktgh/edparyecdoohsaabigteatgiroonu,ps. TSGihrgeoniuf1pi.sc6anaatdnlmydirn3ei.sd2tumecgreieddkgb1io.dd6ayymwgde/oikgsgha/tgdaegysaiaonfnsdPfoFhriOgtShheecreondntotisraegpersecoofhatbhietatetsitonartpiecrlieodh.ad s2ingdnifbiocdanytlwyeirgehdtucgeadinbsodduyriwnegigghetstaotnioDn.L The 3.2 1. Body mign/ukegd/dtaoyreddouscaegebogdrywoeuipghhatds weight gains tended to be wreeidguhctedlosins tohcecu0r.4remdgi/nkgth/eda1y.6domsg/akgge/gdraoyudpoosnagDeLsgr1outpo dosage group was precluded from further evaluatio b 4, when (the 3.2 msiagknigf/idcaanyt before DL 2) n ecause ail pups died 000522 418-008:PAGE 1-6 33A..b22sommlggu/ltkkegga/ldndadayyrddelooasstaaivggeeefgegrerdoouucppossnscduournmitpnitgniuotehndevtpaorlehucaeosvheawberiretedaturiceoednupceerdioidn. thTehe1.61.a6nadnd Mfeaetdercnoanlsfuemepdticoonnsvuamlputeisoenarvlayluinesthteengedsetdattioobneperreidodu.cedAdbisanobtlshuoetleu0t.a4enmad/nrdkealraletldiavateiyve 3fdo.or2stamhgge/eeknggtr/iordueapylaadcnotdastaiwgoenerpegerrsoiiugondpi,fwiaacnasdntplarytermceoldusudtceiedndtferirnovmatlhesevwa1il.tu6haimtnigot/hnkigsb/epdeariyodd.osaTghee group no surviving pups after DL 1. cause there were pcDyaocrslaiamngegetisenrotsfhewthe1er5eterusanttsaafrpfteeirccltdeeodassbayhgietghhgerao3s.u2p3.mt2gha/mtkgg/w/kedgra/eydeadvyaoldsuiaadgtenedo.otfafPMfFaeOtcStin,egstarnodusfertity All necropsy observations were considered unrelated to treatment, elTimhttbeerrreyaovweserraaetgeCnasoefsboairorcleooagrnip-coasrlelacytliiumotepnaoi,rntgiamnoptnlaonDrtGasttai1to0i,nsst,icvalilayblseigenimfbircaynotsdiofrfneornevniceasblien the ogTfrhoieumppd.luarananttaiotobinsoeonrfsvgiaettseistoapnteiarosndsowacamisawtsaeisdgnwisfiiigtcnhainfptirlceyainmrtpleyldaurncetedadutciieondnt,lhoerses3su.[l2tthmieng/akvge/rdaagye dnousmabgeer 3r.e2dumcge/dkgl/itdtearysidzeo]s. agPeugproviuapbsi.lityRewfalsecstiignngiftihceasntelyefrfeecdtusc,etdheinvtiahbegil1it.ny6aaansniddgnificantly dglroacostuaaptgsie,ongtrihonedulipac,cetsaastwieaolrnseoinswdiegenrxiefwiatchasentalalyvseorresadigugencisefdifcoiarnnsttluhyrevri1ev.di6nugacnepddupi3ns.2tihnmeg/1.k6g/mdga/ykgd/odsaayge dosage group group on DL 1 beginning on DL 4 posteulling, and in the 3.2 the 1.6 mg/kg/day mg/kgiday dosage hgAraodvuoepsraaegddemu-icdneeidsptepenurdpeedwnettihpgeahtttetssetronanrotDifcLlree.4du(Tcphreeed- 0pa.un1pdmbpgoo/sdktycgu/wldleaiinyggh)dt,oswcaaogsmepegvridoeunpttiennedaecdhfo Pcounptrbooldgyrowuepigvhatlsueo.n TDhLes 01.4thmrgo/ukggh/4da(ypodstocsuallgiengg)r,ocuopmtpeanrdeedd ttoaorthehaedvcetoonrtterhdoeulced vdgaorrosiuaatpgieovnagsl.ruoe.uTphsHeow1we.er6veemrgs,t/aktngios/tndiecaaylolfdyotsshieagngdieiffigfcearnroteunapcnehdsadimnastyihgenrie0fp.i1rceaanstnelndyt0n.4ormmga/lkgbi/odlaogyical swiegingihfitcsanotnlyalrlewdeuicgehdinpgupdabyso.dy Twheeig3h.t2omng/DkLgl1d(any0 dpouspasgesugrrvoivurepedhdtauocdseaudbpsuepqubeondty scheduled weighings). pColtineinctailalanredduncetciroonpsinymoabtseerrnvaalticoanrseaoscscoucriraetdedinwtihteh 3r.e2dumcge/dkgp/udpayvidaobislatgyeangdroup 000523 418-008:PAGE I-7 nrAuedrmvsoievnregsde,(tcawlnoindoicfoaltnheoebshseaedrlviptaeutripsosnhstahdiatnpcwulepudrseeftrnhoortmeenwehlsiittceedhr)s.twhieAtphlpaucepnsttahhatadwenroet nboeten acanndniavaclainznaitbiaolnizweadsfaolrseoliemvbidinenatnointhtehirspdupo)s.agIet wgarsounpopt(pmapiorssessniitnbglmetaatitleoridnneaotlneermpiunpg Whether these occurred before or after these pups had died. PpNuueppcssr. owpiTsthhyenob3o.s2emirmlvgka/tikin gotnihsdeaisyntpdouompsasacghtehiagntrtwoheuerpe1a.6slstaoinlhdbao3dr.2eovrmifgdo/euknngcd/eddaoefyaddoisnadgiceatgerdoumpany mthaetseernpaulpcsanhnaidbamliiszsatiinognhaitndnleicmrbospsayndo/fosrtplolrotoiomnaonfdtfheoutnaidl.deaidncpruepass.edNine of wfRiiegtvhhetribsnogidbwylaewsediedglehaltyasyoecidncurirenrftlehedxe ia1nn.t6dheapnh0yd.s4i3c.aa2nldmdg1e/.vk6egml/god/pakmyged/nodtsatayhgadetoasgrraegehiggrholuypcso.rreSluartfeadce eiovaeilr orpigmhet nwterefordeplianyneaduninfotlhdein1g..6 emyge/koap/ednianygd,oascaoguestgircosutpa(rontuoprs3e..f2leTxmhoaernktdgi/amdebaioylfy d0 o(sVeahgieclger).ou0p.1,pu0p.s4 waenrde1e.6vamlgu/aktge/ddfaoyr tdhoiss apgaeragmreoteurp)s. All had response when testing was done on DL 21 (end of lactatio live the pups pupil in the constriction n) ggUerpnooeunrpawtpieuoapnnsipwnuegprsoefisntmhtaehlellit(1t.te6rhsemy(gDhkLagd2/1dg)a,aiyanedddoecs2ia0sgi%eonglrewosausspw.emiaTgdhhete dr1ue.rg6amrgd/inkgg/tdhaeyFd1osage the rats cwoentrreolnoptupssui)taabnlde ifnortfhuertohpeirnieoxnpeorfimtehnetaatttieonnd.ingIt laboratoriynvgetlearcitnaatriionant,hathne had been determined that tCinhofenotdrimonasutaeigodnedaoonfsdi1n.wg6omoufgl/d1k.sg6u/mbdgjae/ycktgw/tadhseaaypumlpeovssetltowlihakdiedcliyhtwipoornuoallddudcnisoettdrercsesos.umlpt oinunadnytoaxdidctiyt.onal c1o:n6smualtlaktgilodnawyitdhosthaegeSpgornosuoprFl1eagdenteortahteiodnecpiuspiso.n not to Therefore, continue the Only the 0 (Vehicle), 0.1 and 0.4 mglkg/day dosage group pups were continued to a second generation. B3. F1 Generation Male Rats: wNeoredeaaftihrsibuintetdhetoFP1FgOeSn.eraAtlliocnlinmiacalleorbastesravtat0i.1onasnwde0r.e4 the test article. mg/kg/day considered dosages unrelated to wgTerhioeguhp0t.1opneardnadwye0e.1k4pmtoghs/atnkwget/ahdneaicynogndtoarsnoaldgggereongeurrpoaulrpaltsys.tgeaRniendfeelddecottoninewgeoitrhgithswlopesgsrathmasnltehses cboondtyrol pgraienc,ohwaebiigthattigoaninpseriinotdhwee0r.e1 and 0.4 mg/kg/day dosage attern of groups for the weight 98.4% and 97.6% of the control group value, 000524 418-008:PAGE I-8 deaspyec1 itvoetleyr.minBaotdioynwweeirghet9g8a.i5n%s ianntdhe960..16%anodf respectively. However, none of these body we 0.4 the cmogn/tkrogl/dgaryoudposvaalgvee.groups for statistically significant from control values. ight gain differences were Cw`Aoebnrsseoulsmuiptgetniiffoeinecadvntaclloyunersseudmtupectneiddoendontvoaplbouesetsrwefeodaruntcihenedg0id.n1atyhasend10.t01.h4romugg/hkg8./dRaeyladtoisvaegfeegerdoups groups during the first week of intubation but none wearneds0ta.t4ismtgic/akllgy/dsiagynidfoicsaangte DdDiaooylsoaoggficepasrlelpoyuftitimhapelortsteesaptnatraradttiiicfoleenraeisnncthehiseghiFn1atsghee0n.v4earlmaugte/iskongf/omrdaallyeeadirdatsn.otTahfefrecetwtehreaevenorage pelaovrnaagld-utiaegtrmem.d rbeytepnetrifonoromrarnecsepionnasepainshsiibvietioanvoinidtahnecFe1ogrewrnnaeitrneag,trimsoahnzormeta-plteeerrfrmaotrrse,mtaaenqn.ctieon, aDnodsafegretisloitfy ptahreatmeesttearrstieclvealausahtiegdhiansth0e.4F1mgg/eknge/rdaatyiodnidmanloet araftfee.ct anymating ouAlcnlcruenlrearcteredodpisntoytthohebesaetbressvotaltaurittoeincoslreinrbeetlchaaetuisvFee.vgaelNnueoersasttfaiotroinstthimecaawllleeyisraitgsniwfiecraentcdoinfsfiedreerneceds sttteeasnttidisset,dicsatelomlyibnesailrgnevidefusicciacenldtesif,nrtrihgeht0.e1piadniddy0m.i4s mogr/pkrgo/stdaatye.dogTsheatrgsmeionfgartloheubporsdigyhbtuwteoriwgelherftets not om control values. B4. F1 Generation Female Rats/F2 Generation Litters: gANleolstadadetvaietornhssaenicndltilnhiaeccatFla1toibgosneenpreevrraaittoiidoosnnswfetehrmaeatlcoeocncrsautirsdreewrdeerddeuruaintnrtgerlitbhuetepdrteocoPhFabOiSt;ation, ated to the test article, Body weights postweaning. tended to Matemal Reriod were unaffected bboedryewdeuicgehdtsinatnhde by dosages of the b0.o4dymgw/ekigg/hdtagyaidnossadugreinggrotuhpe test articie on day 1 gestation 0S.ig4nmifgi/ckagnt/dmaaytedronsaalgbeogdryouwpe.ight loss occurred on as DLs 1 high to 4 as in 0.4 the mg/kg/day. crToehdneuscu0t.mi4potnmigio/nnkagbo/sndoladuyatydesofs1eaetgode8coopfnostshutemweptatensitionnagr.taincldAebawsaotlseuntadesesnaoccyiaftoerdrewidtuhcaedsrieglnaitfiivceanfteed tfhoenstuersmtpatritoicnlevaalsuehisgdhuarisn0g.4thmegg/eksgt/adtaiyo.n period were nd relative unaffected feed by dosages The absolute and relative mates! of 00S25 418-008:PAGE 1-9 feed consumption dosage group. values were reduced during lactation in the 0.4 mgkglday dDaoysaogfevsagoifnatlhepatetsetnacrytiicnlethaes Fh1igghenaesr0a.t4iomngf/ekmga/ldeayradtsi.d biologically important differences in the values for nTohtearfefewcterteheanvoerage eplavoranagld-uitagetmre.md rbeytepnetriofnoromrarnecsepionnsaepainshsiibviteioanvoinidtahnecFe1loegraewrnnaeitrnaegt,rimsoanhzofeerpmtearelfreorrremattaesnn,ctaeis.o,n, aDnodsafgeretislitoyfptahreatmeesttearrstieclvealausahtiegdhiansth0e.4F1mgg/eknge/rdaatyiodnidfenmoatlaeffreactst. anymating All necropsy unrelated to tohbesetersvtataritoicnlse,in the F1 generation female rats were considered 2dP5orsefagegnmaeanlgceryoruaoptcssu,arsrresesidpgencietnid2ve2tloy(.c9o5h.Aa6lb%i)tp,arte2ig1onna(n8int4.td0ha%em)0sa(ndVdeelh2iicv4le()9,6.00.)1 oafntdhe0.243,mg2/5kagnidday pirnedveixowuassgecnoemraptairoanb,ltehearcerowsassthaettehnrdeeendcoysfaogreregdruoucpesd.erpeuAdpslivotiecarcbsiu.lrtryTehdineitngheteshteetios 0:4 mglkglday iinocsraegaesegdr,ouapn.d dtohseanguemgbreorupo.f pTuhpedneuatmhbserafotferdDaLms2 wailtsho As also occurred in the previous gene swteirlbeorinncpruepassewdais this gtporesontudapeuldvlaitlnogub)esea.nrdePduDupcLeb7d.o inBythDeL0.114,mgb/okdgy/dwaeyigdhotssawgeeregrrcoaoutmpipooann,rapDbLul.pe4bto(opdcryoen-wtaernoidlghts fbeedurcetdiuocnesd,prtehsoeungthonnodtDyLsiswgen7iifgaihcntadsntli1y4n.,thoeNno0D.cL4lsimni4gc/atklhgor/rodunageyhcdr2oo1,spsawygietohbgssreiogrunvipaftiticeoanndted to F2 0.4 generation pups mg/kgiday. were attributable to dosages of the tes article ac ns high in as the 000526 Cc. Conclusions 418-008:PAGE I-10 obhOsinegrhtveharebdlboea-sseaifsgfeeoscfttc-haleuesvseeeldda(trNaeO,dEutcLht)eioFonfosP(iFgneObneoSrdayitsiw0oe.n1imgmhagtt/egkragni/anldaaaynndd(0r.pe4adt`uemrcgne/adklgf/nodoa-ey and `consumption values). The Fo generation reproductive NOEL is greater than 3.2 mg/kg/day; no effects ihoningthmheaertFid1nogs,geafngeeretrsialitctiyaoounrseoefdsftsprproreuiisnmgpclyiscaln0it.na4gtmioogcn/cuklrgor/sesdda.ayndT(1h.re6edmNugcO/tEkigLon/sdfoairynvliaianbtiedlristiyzae,ndpugprowth viability, growth and survival). T(0h4e mFg1/gkegn/edraaytidoonsmagaetecmaaulseadndrepdautcetrinoanls consumption values). NOEL of PFOS in body weight is 0.1 mg/kg/day gain and reduced feed aT0.nh4demgFgr1/okwggte/hndeairyna;tthineoonFe2rfefgpeerctnosdeurocanttiimvoaentioNfnOfgsEpoLrrinifsegrgtiirsiet0ay.t1oecrmcgtu/hrrakengd/.adadTyohseagNeOoEfL forviability v(i0a.b4ilmitgyi,kgar/odwatyh daonsdsaugrevicvaalu)s.ed stilbirths and reductions in fter size, pup ; Eteve liidredf Christian, Ph.D. , Fellow, ATS Executive Director of Research Date Le h< se=Yr Alan M. Hoberman, Ph.D. . DABT Director of Research Date 2 74p1d0 u 90 R: nd G. York, PJ, DABT Date Associate Director of Resedrch and Study Director 600527 n SCRIPTIONOF TEST PROCEDUI 418-008:PAGE II-1 A. CondofuStcudty: AA. Sponsor: 3MiMnnTeosxoitcaol5o5g1y4S4e-r1vi0c0e0s, 3M Center, Building 220-2E-02, St. Paul, A2. Testing Facility: APregnnussyRlveasneiaarc1h9L0a4b4o-r1at2o9r7ies, Inc., 905 Sheehy Drive, Building A, Horsham, A3. StudyNumber: 418-008 A4. Sponsor's Study Number: 6295.9 AS. Purposeofthe Study: TPhFeOSputrrpeoastemeonftthoifsCsrtlud:yCDwaBsRtoVAteFs/tPflourstoximcalefefeacntds/fdeimsatluerbraantcsesberfeosruleting from ceovhaalbuiattaetiIoCnHtHharorumgohnimsateidngT,ripgaersttiatteiGouniadenldinlaectsattaigone.s AThtihsrsotuugdhyFwoafstdheesigned to irrametppslr,aonodtnuactttihioevne,depgvreoestclaeotspismonea,nntpdaordfteuttrhietecitoonfe,ffsflpearccittnsagtoionofnttahhenedetsrmtearatoteeurdsnacmylacllbeee,haatnvudbiaoflremtiranalfnesmpaolret., aspnedrpmermmaitturdaettieocnt)iotnhaotf mfuanyctniootnableefdfeetcetcst(eed..b,y ehfifsetcotlsogoincalliebixdaomoirnaetpiiodniesdyroamftaslm, ale praetriroedprmoaduyctbievedeolragaynesd. inBtehceaoufsfseprmianngi,foebssteartviaotnsioonfsewfefercetscoinntdiuncueedddtuhrriongugthhis production of F2 generation litters. AS. Study Design: Awermoediufsiecdataiosnaofbatshiesrfeoqrutihreemsetnutdsy of U.S. design Food and Drug Administration (FDA) 000528 AZ. Regulatory Compliance: 418-008:PAGE Il-2 (TGhLePs)truedgyuwlaatsiocnosondfutchteedU.iSn. cFomopoldiaanncdeDwriutgh AtdhmeiGniosotdratLiaobnor(aFtoDrAy)?Pratchteice CJoampamnuensietyMi(niEsEtCry).of Health There waenrdeWneolfsairgeni(fiMcaHnWt )deviaantdiotnhse fEruormotpheeaGnLEPconomic drUneoigctuulfmaitenindoitnnesgdsthdaaentrdaifvhfeaedvcefterbdoemtehtneheqpuriaonlvsiiptdeyecodtritoionnsttehdgeruirStiytonugfdttyhhDeeicrseotcnutddoyur.catnQoudfaltthihitesyTsAetsusduyraarnece Facility Management sting AB. Ownership of the Study: TtihsesuSepsoanrseotrheowpnrospetrhteyostfudtyh.e All raw data, Sponsor. analyses, reports and all preserved AS. Study Monitor: Marvin T. Case, D.V.M., Ph.D. A.10. Alternate Study Monitor: AndrewM. Seacat, Ph.D. A.11. StudyDirector: Raymond G. York, Ph.D., DABT (Associate Director of Research) A.12. Technical Performance: John F. Kristen lBaanmdeotlta,SBh.eSr.er(,DiBr.eS.cto(rRoefseLaarbcohraAtsosroyciOapteer/aFteitoanls)Evaluation) Joseph Sharon W. Lech, Adamski B.S. (Team (Laboratory TLeecahdneirci~anG)eneral Laboratory) A13. Report Preparation Raymond G. York, Michelle R. Rzaca, Ph.D., DABT B.S. (Study Coordinator) HEreiindiHMa.gaGnr,eeBn.A,. M.(SD.ata(SMtaudnyagCoeomredinntatSoprec)ialist) Karen G. Parker, A.A. (Report Administrator) 000529 A.14. ReportReview: 418-008:PAGE I1-3 AMliladnreMd. SH.oCbherrimstaina,n,PPhh..DD..,,DFAelBlTow,(DAirTeScto(rExoefcRuetsievearDcihr)ector of Research) A.15. DateProtocolSigned: 21 May 1988 A16. Dates of Technical Performance: Eo Generation Male Rat DRaotsaArgreivPaelrDiaotde(42 days before cohabitation, ptehrriooudghanad1u4n-tidlaysaccroihfaibciet)ation Scheduled Sacrifice 12 MAY 08 26 MAY 98 - 30 JUL 98 31JUL98 Eo Generation FemaleRats RDaotsaArgreivPalerDiaotde- Female Rats Assigned to 12 MAY 98 cCoaheasbairteaatnio-nSeacntdiocnoinntgin(u4i2ngdtahyrsoubegfhoDreG* 9) Dosage Period -- Female Rats Assigned to 26 MAY 98 - 17 JuL 98 cNoahtaubriatlatDieolnivtehrryo[u4g2h days before DG 24 (rats that did not deliver a litter) a litter) or DL*20 Dosage Period Estrous (rats that delivered 26 MAY 98 - 30 AUG 98 Cycle Evaluation 09 JUN 98 - 06 JUL 98 2 b. DGis DLis used used as as an abbreviation abbreviation for for day doafylaocfta(tpiorne.sumed) gestation 000530 (Fo Generation Female Rats Continued) Cohabitation Male 1 Period Male 2 DNaGtur1a0lCDaeelsiavreeraynP-eSreicotdi(onDiLn1g) DG 25 Sacrifice a litter) (rats that did not deliver DsLel2e1ctSeadcrfiofricceon(tdianmusedasntdudpyu)ps not E1 Generation Rats 418-008:PAGE ll-q 06 13 JUL JUL 98 98 P P---M12M 30 JJUULL9988 AM AM 2187JJUULL9888--1300AJUULG 9988 01AUG 88-07 AUG 98 17 AUG 98 - 30 AUG 98 Dosage Dosage Period Period (Male Rats) (Female Rats) Passive Avoidance Testing 19 19 AUG AUG 98 98 - 16 27 NOV DEC 98 98 WCoahtaebritmaatzieonTePsetriinogd (Initiated when the rats are 20 07 AUG OCT 98 - 10 98-19 SEP OCT 98 98 aMpaplreox1imately 90 days of age) Male 2 02 NOV 98 PM -- 09 NOV 98 AM NMaatlueraRlatDseliSvacerriyfPiecerdiod (DL 1) 09 NOV 98 PM - 16 NOV 17 NOV 98 98 AM 24 NOV 98 - 08 DEC 98 DL 21 Sacrifice 14 DEC 98 - 28 DEC 98 A17. Records Maintained: vITnechh.eicoAlrenigycionpamrlperoseenpreovrntet,dsrtaairswesdureaesttaaairaneneddreritneasitneheredvaeirncshtaihmvepelaserocsfoAfrtghuesbRueksetaesrtchaLratibcolreaatnodries, aSdteocteihdeyeeTaetrhsetaiifrtnefgirnFmaalaciidlliiisnpogstithieond.rafAtllfiunanlurseepdortt,esatfatrelriwclhheiiscvuhesstpoiefmnetshitehoenTseSspwtoeinrngsedoiFrascwciilailrtydefodr Monitor. ty. Unused bulk test article was returned to the Study B. TestArticle Information: B.1. Description: PFOS - an off-white powder B2. LotNumber: 217 (Expiration Date: May 2000) 000531 B.3. Date Received andStorage Conditions: 418-008:PAGE II-5 TPrheeptaersetdarstuisclpeewnsaisonrescweeivreedsotnor2e0d aMtaryoo19m9t8e,mapnedrasttuorreedovaetrnriogohtm.temperature. BA. cial Handling Instructions: Standard respirator saanfdetsyapferteycaguotgigolness)(uwseerofeptraokteenctwihveenclhoathnidnig,nggltohveesb,uldkustte-stmiasrtticle and prepared suspensions. B.S. Analysis of Activity: Information article is on regarding the purity, file with the Sponsor. identity, strength and composition of the test C. Vehicle Information: C.1. Description: 0.5% (RO. Tween 80 in reverse deionized water). osmosis membrane processed deionized water C.2. LotNumbers: MO3HOS, K03737, M29477 and L06662 C.3. Dates Received and Storage Conditions: 1T5heAuTgwuesetn199870 (wLaotsKr0e3c7e3i7v)e,d f2r2omMaJ.yT.1B9a9k8er(,LotPhMiOli3pHsObSu)rg,, 1N7eSwepJetresmebye,ron1998 (Lot (Lot LM02G9G46727)),,a3ndDewcaesmbsteorre1d9a9t8r(oLootm MO3HOS) and temperature. 1 September 1998 R.O. deionized water is arvoaoimlatbelmepferraotmuraec.onTtihneuoveuhsicsloeurwcaesatprtehpeaTreesdtiwnegekFalcyilaintyd and is maintained stored at room at temperature. 000532 ca. i indling ictions: 418-008:PAGE II.6 Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles handling the vehicle. or safety glasses and a face-shield) were taken when C.5. Analysis of Purity: Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. D. est Article Preparation: Suspensions of PFOS were prepared daily one day prior to the day of dosing at concentrations of 0, 0.02, 0.08, 0.32 and 0.64 mg/mL. The test article was considered 100% pure for the purpose of dosage calculations. D.1 oh Sample em Infor tion: i HAE ws vance Cement Reserve ET Rink Ee ose ged SEER Stability data for prepared formulations bracketing the range of concentrations are on file with the Sponsor. The Sponsor confirmed a 48-hour stability on the test article in 0.5% Tween 80 solutions. a. SSeTeAANPDPAERNDDOIXPEGRA(TDIEVNIGATPIROONCSEFDRUORMESTHOEFPTRHOETTOECSOTLINAGNFDACTIHLEITY), item 1 000533 D2. Analytical Results: 418-008:PAGE II.7 Results of time of the the concentration and writing of this report. homogeneity analyses were not available at the. E. TestSystem: El. Species: Rat E2. Strain: Cr:CDBR VAF/Plus (Sprague-Dawley) E.3. Supplier(Source): Charles River Laboratories, Inc., Raleigh, North Carolina Ed. Sex: Male and Female ES. Rationale for Test System: The Crl.CDBR System because: VAF/Plus (Sprague-Dawley) 1) this strain of rat has been rat was selected demonstrated to as be the Test sensitive to irnedpursotdruyctfiorvereapnrdodduecvteilveopamnedntdaelvetloxoipnmsenatnadlhtaosxicbietey nevwaildueatliyounss;ed2)thhrisotuogrhicoault pdhaatramaancodleoxgpiecarlileyncaecteixviestinatthtehespTeecsiteinsgaFnadcilstirtayi'n."; and 3) the test article is ES. TestSystem Data: Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight Weight (g) (g) on at the Day after Arrival Study Assignment Male Rats 195 12 MAR 98 62 days 282-338 334-396 Female Rats 205 12 MAR 98 62 days 172-239 209-242 000534 E7. Method of Randomization: 418-008:PAGE 11-8 E.7.a. Fo Generation Rats Upon arrival, Fo generation rats were of computer-generated random units. aAsfsteirgnaecdcltiomaitnidoinv,idmuaallehoaunsdinfgemoanlethreatbsasis wreecroerdseeldedcutreidnfgoracsctluidmyatoinont.heRabatssiwseorfephaysssiicganleadptpoefairveandcoesaagned gbrooduypsweights g(eGnreoruaptsed| t(hwreoiugghht-Vo)r,d3e5rerda)tsrapnedrosmeixzapteirondoprsoacgeedugrreo.up, using a computer- Tashseigfinrsetdtteon Cfaeemsaalreearnat-ssepcetridoonsinaggeongrDoGup1w0i.thTahecornefmiarimneidngmartaitsngwedrateepweermrietted to naturally deliver litters. Aphtaarbmlaecoofkirnaentdiocmsaumniptlsewcaoslluecsteidontoeiatshserigant fsicvheerdautlsepdersagcrriofuipcetoafter completion odfeltihveercaohlaibttietra)toirononpeDrLio2d1(m(faelmearlaetsrastisrianlglloiwtetedrstwoitnahtduraamllsy adlelliovweerdatolitntaert).urally E.7.b. FA/F2 Generation Pups On DL 4, a litters were table of reduced random to eight units was used to select the pups to pups each. Whenever possible, the be culled and same number of male and female pups per litter were continued on study. At weaning of used to select the 25 F'1 generation pups male and 25 female on DL 21, a table of random units was. pups in each of Groups I, ll and Ii, resulting in evaluation. a total of At least 150 one mFa1legepnuepraatniodnornaetsf(e7m5alpeerpsuepx)pecrhloistteernwfeorrecosnetliecntueedd. Tcohnetrienuweedreonnothseursvtiuvdiyngbepcuapussien oGfroseuvpeVreatfotxeircDeLff2e;ctpsuipnsthineGpruopusp(IdVeawtehraenndot retarded growth) during lactation. This the study veterinarian and the Sponsor. decision was made in consultation with ES. Syos fIdetntife icatm ion: E.8.a. Fo Generation aRnatdsTwaegreCo.p,erImnac.n,enNot.lyMiSdePnTtif2i0e1d0u1s)i.ngMMaolenealnd sfeelmfa-plieerrcaitnsgweearretaagsssi(gGneeyd Band tneummpboerrasrywhneumnbaesrssiganterdecteoipttheanstdudgiyvbeenfourneiqaudemipneirstmraanteionnt oifdetnhteiffiicrasttidonosage of the test article. 000535 E.8.b. E1/F2 Generation Pups and Rat 418-008:PAGE II-9 rePvauacplhsua1wteerdgeeinnneotrteatriimndosinvoirfdauttahsleelylfietictdeterenbtdoifftioherdbcedofunortriengalnadctaaftieorn;cuallllipnagroanmeDtLer4s. were Awteaning, Monel self-piercing ear tag. inued observation was identified wits F. Husbandry: FA. Research Facility Registration: SDA et seq. Registration No. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 F.2. Study Rooms: 12Th0he0al%sltwfuardeyysahrnoadoimitsnhdawetepheraneddembnaetilenytnasipunapespdslieuednddtwehirrtochuoagnhdmi9iti9noinmsuomf opofstietnivcehaainrfgleosw relativeto per hour of hRtueommoipdmeirttayetmwupraeesraatntadurrgheumwiadsittyarwgeerteedmoatni6t4orFedtoco7n9stFan.(t9l17y8%tChHrEtooPu2gA6h-ofCui)etr;st.hreesltaRtueidveye.m AND RELATIVE HeUtMedIDatIT3Y0%REtPoO7R0T%.S).See APPENDIX H (TEMPERATURE F3. Housing: ACarceaagnedsiUzseesoafnLdabhooruastionrgycAonndiimtailosns.were in compliance with the Guide for the F.3.2. Fo Generation Rats/F1 Generation Litters CFoaohgagebesinteeartxaicotenip,otnedaruacrthisnpwgaeitrrheeoficnordhaitavsbiidwtuaaatslilyohnohuaosnuedsdepdionsintthpseataritnulmespsersitoedesl.wiDruer-ibnogttomed iilanntdaeirvcitdohumaalnmloyDhGou20s,edFoingneensetriantgiobnoxfeesm.alEearcahtsdaasmsiagmnnaedldedteorlaintv'aestrcueardgael.idteelrBiewvgeorirynenwihenorguesneod n nesting box during the postpartum period. F.3.b. 1 Generation Rats/F2 Generation Litters acAofnhtdaebriinwtdeaiatviniodinun,agl,lhyotuhhseoeuFds1eidgneapnfaetirerarstc(iooohnnaebriamttaastiewoenrr.aet Tpinehdreivfsiedmuaallleyrhato)udsuerdinbgecfoohraebitation, 2s F1 dgeesncerriabteidonfofretmhaeleForagtsenweerraetiionndirvaitds.ualBlyeghionunsienadgmnieno type of caging nleastteirntghabnoxDesG, was used 0, Each dam 000536 418-008:PAGE II-10 and delivered period. liter were housed in a common nesting box during thepostpartum F4. Lighting: ifinghatu:t1o2m-ahtoiucraslldya.rcko,nwtirtahlleeadcfhludoarrekscpeenrtioidghbtegciycnlneinwgaast m1a9i0n0tahionuerdsaEtS1T2.-hours F.5. Sanitization: aeCsargneeeccpehasansnaglrieyndetroaspkwpeereropxeitcmhhaeatenralgtyesdedvraeyprpayrnoodtxhicemlreaatwnee.leyk.thrBeeedtdiimnegs weaaschchwaenegkeCdaagessen F6. Feed: RNuattrsitwioenreIngteirvneantiaondall,itInucm.,aSct.ceLsosuitso, CMeirstsiofuireid)RiondienndtivDidiueatlf#e5e0d0e2r (PMI F.7. Feed Analysis: Afjenovamellysesxeepxseccewteeedrdienngruotturhtiiteinoemnlaaylxpireemrqfuuomirrmceeomndecnbetnysttrwhaeetrifeoenedsdetfsoeurcpptcleeiretri.fiedNofeceodnotramdienvainattsioant of the resuitsof the feed analyses are available in d the by these raw data, analyses Copies either the Study the feed that was kDniroewcntotronionrtetrhfeerSepwointshotrhewaressualwtas roef of any agent this study. present in F.8. Water: JmLroeacmtaeblrriwnaagntaeecrc(tReh.saOst.shwyaasdttebere)mewannadps/roaorvcaeiisnldsaiebvldiedbutayolptwhaaestsreaatgseatdhrloibuigthumafrreovmerasneaoustmoomsaitisc the processed water as a bacteriostat. r bottles. Chlorine was added to F.9. Water Analysis: QcToohnnettanpmlriyoncfaeotrsispoeonsd(swiLbaalntecearbsaitcsetarenrLiaaallbyozcreoadntottrawimieicsne,ataIinncon.nu,a(lLAlanynaclfayostrtipecoars,siPbelnenscyhlevmainciaal) and Chalfont, available Pennsylvania). in the raw data. Copiesof the resuits of the l Laboratories, Ing. water analyses are 000537 418-008:PAGE [1-11 Neither the Study the water that was Director known tnooirnttehrefeSrpeownistohrthweasresauwlatsreofotfhaisnystaugdeynt present in F.10. Bedding: GBreodu-p0,'cM0abusmewea,sOuhisoe)d. as nesting material (The Andersons Industrial Products F.11. Bedding Analysis: rcNeoesinuttlhatesmriontfahtenhtiSsspslotinukdeslyoy.rtonAonbraeltpyhrseeeSssetnuftodryipnDotisrhseeicbbtloeerdcdwoiannstgaatmwhiaantrweooufldaniyntpeortfeenrteiawlith the annually and documented in the raw data. ation are conducted G. Methods: G1. Dosage Administration: I| OGosoawse|| (mDgorsgaigceay|| Con(cmenatirmat)ion | oDos(aghesVo)li ume|| CNauNmurbmeebresroofftFos|| GNeurmmbaeirnoftrFrsf eTloovo eTT owe [ --er 2 Se-- x --ersex | C[oo [e eT [oene [TT Le[oe Tom[0 The test aticie was considered 100% pure for the purpose of dosage calculations 000538 G.2. signed Rat Numbers: 418-008:PAGE I-12 C [ T DosageT GT ro[ up Woo eo ovensTTewveFaoorww nsseo1 m |orovwawon es o|] r] aerse]o|] Cy aavmen[wwawmos[0 h10o2sstaetrheare seregvoterermrseroroenmnere G.3. Rationale for DosageSelection: cDoonsdaugcetsedwewritehstehleecttesetd abrtyictleh.e Sponsor on the basis of previous studies G.4. Route ofAdministration: Oral (gavage) G.5. Rationale for Route ofAdministration: The oral (gavage) route was selected for use because: 1) in comparison with the dietary route, the exact dosage can be accurately of the possible routes of human`exposure. administered; and 2) itis one G.6. Frequencyof Administration: The Fo generation male rats were given appropriate dosages of the testarticle monacxeimdauimlyobfeg1i4ndnaiynsg)4a2nddacyosntbienfuoirneg cohabitation through the (which continued for day before sacrifice. a The Fo generation female rats were given the appropriate dosages of the test article beginning 42 days before 14 days) and continuing through cohabitation (which DG 9 (rats as continued for a maximum of (DraGts2t4ha(tradtisdadsesliigvnereda to natural delivery that signed to Caesarean-sectioning), did not deliver a litter), or DL 20 litter). The dosage volume (5 mlkg) was adjusted daily aopnprthoexibmaastieslyoftthheesmaomset triemceenetlaychrdeacyo'r.ded body weight and given at a. See APPENDIX G, item 2. 000539 418-008:PAGE I1-13 pcDaaarntnmuirsbiatilinoinzt,ahteiinpoonrroodcfeertshsteooppfurpdesec.lliuvdeCreoinpnsgoespqsuuiepbnlstelwdyei,rsersuopntomiteodndoosfemdatunetmilaclombpelheatviioonr aofndor one daiy dosage daily dosage. during the delivery period. No ams were dam missed not adminiteres more than ane aFr1tigcleeneorraaltliyo(ngmaavlaegea)ndbefgeimnanliengraotns through the day before sacrifice. F2 wdearye1 gpiovsetnwaepapnrionpgri(aDteL generation pups we dosages 22)* and of the test continuing gteessttaatritiocnle(,inbuuttemraoyexhpaovseurbee)eonr possibly exposed via maternal milk to the treestnaorttidcilreecdtulryinggiven during the lactation period, fhe G.7. Length of Study: Approximately 7 months G.8. Method of Study Performance: G.8.a FoGenerationRats oAplnelrciFeooddsugreoinfnegtrhatethiseotnuadcryca.ltismRwaatetirsoenwoepbersreeioradvlseaodnodfborfsoevrricavlbeiindlitcfyaolratogbelseneaersrtvaalttwiaipcepedaairlayndcueriantgloaialst taefstteradrtoicslae,geabaonrtdioonns,thperedmaaytsuarceridfeilcievde.ries, prior to and approonxsimoafteeflfyecotnseofhtohuer `BaocdclyimwaetiigohntpserfioordF,owegeenkelryadtuiroinngmatlhee rdaotssawgeereperreicoodradnedd aatt consumption values for male rats were recorded weekly du least once sacrifice. during Feed the period ring the dosage. tBhoedayccwleiimgahttisonfopreFrioodg,enweereaktliyontofecomhaalbeitraattisonw.erdeairlyecdourrdiendg on DLs 1, 4. 7, 10 and 14 (rats assigned to natural delive at least once the gestation during period, gFeesetdatcioonnspuemrpitodioanndvaolnueDsLwser1e,4r,e7c,or1d0edanwdee1k4ly(rattoscaoshasbiirgtyna)etdaiontndo, at sacriflos. daily during natural the delivery). expected tFhaetetdhecopnuspusmpwtoiuolnd vbaelguienstwoecroennsoutmreecmoartdeerdnaalftfeereDdL 14, when it was bAgevfatolaurbaeltetiohofenrsotafanredtsotormfotuuhsneitccsyocwhlaaibnsigtubastyeieodnxtapomeirsnieoaldet.citon15offveamgailnealractystpoelrogdyofsoarg2e8gdraoysu,p for S-- S ---------- a. See APPENDIX G, item 3 000540 418-008:PAGE I-14 cWoihtahbiintaetaicohn,doonseagmealgeroruapt,pceornfseemcaulteivreato. rdTehrewcaoshaubsiteadtitoonaspesriigond maximum of 14 days. During cohabitation, all female ra rats to consisted of a cunotpiulloabtsoerryvaptliugoninofsistpue(rDmGat0o),zofaolilnowainsgmwehairchoftthheey vwaegrientasalswcseoirngteneeendvtastloauniandtd/eiodvriddaauially cdhooohusasabigintega.tgiroFonuepmw)aelraenedraasrtsesimtgahniaetndeddiadlitnneorctnoahmtaaebtimetaaltwiietohnriantfsotrhtaheafmtiarhsxtaidsmemvuaemtneoddfa(ywsitohfin the same days. seven additional The first ten assigned to Cfaeemsaalreearnat-ssepcetridoonisnaggeongrDoGup 1w0i.thTahecofnefmiarlmeedramtastaisnsgigdnaetde were to rCtaaetesas,waeriermaepnlp-aesnretmcaitttiitooenndisnitgtoenswaeatrunerdalevlxiyaabdmelileniaveendrdflointrotentrsvh.ieanbTulhemebsfeeetrursaaetnss.dwdeTirsheteriebrvueatmliauoinnoifngcofrepmoarrae fcilrisnticpaulpobwsaesrvoabtsieornvsedd)u,rilnigteprarstiuzreit(iaolln,pudpursatdieolnivoefregde)staantdiopnu(pDvGia0bitlotaytthaeetd bdfiaotryh,the lrPieutmeprossvtefhodartvaienaibdtihlieitrmymaweperprseeeadreexidnamwsiatitnleelrbd.orfnoProurvpitsthaalwtistdthaiteludusnbgaestfobtirhraetthi.sniatTinhakleewxleaurmnigsnawteiroen. of the sSthiolrltbloyrna;ftpeurpbsirtwhi.th Elaucnghslitthtaert wfalosatseudbwseerqueecnotnlsyiedxearemdinleivdebdoarlnyafneodrcpotounpshiavdvieearbedidileyd weMaxatasemrrinenacelodrbddeeuhdraivoningorDthLoefs2t11h,e-4dd,aay7m,ps1o4swtaapsnadretv2u1a.mlupaVetraieroidda.dtaiioOlnybsswfehrreovmnedetxhmpeaeptceutrpensdalwmeabreehavior behavior period were recorded, if and when present, on all other days of the termal postpartum TFehretsietyppaarraammeetteerrssiwncelruedeadsstheessfeerdtilfiotry ialnlddexam(spearscseingtnaegde to of natural delivery. matings that PrrUeePssSuu)llt.teednduiimnnbtpehreregbonifartnihcmipoelfsal)ni.tvaegteliisotttneartssii)t,oensn,uinmgdebenexerr(aoplfeocrfocfnesdnpitrtaiignogen opofefrptrhlieetgtdenra(nlcvieeasntdhadtead sduurrviinvgetdhe4 paonsdtp7adratyusm),pearnidod,lavcitaabtiilointyiinnddeixce(spe(rpceerncteangteagoef opfuppasumpbsoarbnnodrtnlhiattttheart survived 21 days) G-8.b FIF2 Generation Pups ~ Preweaning Observations wfDierasryted1aryeofcoolnracdwteahdtiiacofhntea(rlplaolpslutpppusapristnuiman)iawtlaeirsttwedreewrfeeirnieendddiaevlsiidtvuehareleldydawayeniodfgghbreirdoth(apunpd wboadsyawlesiogthhtes omed by the dam). Vdiiteadl bsetfaoturse aitnitbiiarlthexwaamsindaettieornmoifnetdhefolirttpeur pfosrtvhiaabtileiityt.herPaupppsetahraetdeisttihlelrboarpnpoerartheadt 000541 418-008:PAGE II-15 esxtaimbionnedorftohravtitdailesdtabteufsoraet initial birth, eaxsapmrienvaitoiuosnlyodfetshcerilbietde.r for iabilty were P(Eioanscctlhpuadlirinttegurmvwapareisraiteoivdoa.nlsuPfaurtopemsd efinoxrpeevaiccatbheildliitntyeuarrtswilenergaestbcetohwuincteeedaocnhcdeadyidyu.rinPghytshieca2l1-sdiagnys aPnuopmsaloibesse)rviendthaet tphuepssewteirmeesrweceorrederdemoonvceeddfarlaoyvmifotorhre2a1nneddsatgyirnsogspsboosex.txpteaWmrhatelupmnhynDsoietcaadl pnercercolpusdieeddbaynaduteoxlaysmiisnoerdcfaonrnitbhaelcizaautsieonofbydetahteh.damP,upanbyodpyupwefioguhntds recorded on DLs 1 (birth), 4, 7, 14 and 21. dead were was w{Raeebfirlleietxymotaonniditgophrhteydsinidc5uarlsiendcgeovtnehdleso2p(1mf-erdonamtyDppaLorsa1)tmpeatrpeitrnusnmaipneurntifhooedl.Fd1inSgguer(nffearrcoaemtiroignhtpiunpgsreofnlleyx crepfelnexin(gfr(ofmroDmLD1L4)12w)e,raecomuosntiitcorsteadrtulnetirleasllpopnuspes ((i1r0o0m%D)Lin13t)heanlDidtLear2i)rr,ereaigychheteindgthe criterion for the number the specific of pups per test. litter Pupil constriction was with this reflex present evaluated once was recorded. on DL 21: G.8.c. F1 Generation Rats - Postweaning Observations og"fePntoehsretawtsetiauondnyi.nragtRsdaawtyes"rweoebrosebesraevlrastvoieoodnbsfsoerwrevvireaebdilrfieotcryogartednleeedraasbltegtaiwpnipcneeiandrgaaiolyn dDuLri2n2g.alAlllpeFr1iods during article, tahbeoratcicolnism,atpiroenmapteuriroeddaelnidvefroiresclpirniicoarltoobasenrdvaatpipornosxiomnfacetefeflaeytctolsenaoesfthtoohunercfeoasftter dosage and on the day sacrificed wpBeooesdktylwyewaeenixigchnetgpstpfedoruriroFid1nsggaceonnhdearbaaitttisaoatcnirioffneicmaea.nldeFaretaetssdacwrcieofrniecseu.rmepctoirodnedvawleueekslywedruerinrgectohreded caBosohsdaibygintweaedtiigtohont,nsadtfauoirrlayl1dduegrleiinnvgeerrpayrt)ieaosnnudmfeeatmdasglaecersitrfaaittciseo.wn,eFroeenerdDeLccsoornd1s,eud4m,po7ncaendwe1e4kl(ryattso fecorded weekly 10and 14. to cohabitation. on DGS 0, 7. 10, 14, 17 tion values were and 20 and DLs 1. 4, 7, efBeaetcgeihnntdnioionnsgaingaeta 2gp4arsodsuaipyvsweeoarfveoaigtede,satnoecdneefpomarralaledeairgrnmait.nga,nEsdahcoorhnte.ratfteerwmmaarleeternattifornoamnedaclohnlgi-tteerrimn sboetphardaatyesdofbyteastoinnge.-wTeheek pinatsesrivavle; atvhoeicdraitnecreioanpfpoarrlaetaursniscnogntwessiatssetdetdhoenofstaawmofeydoaf-yosr, compartment chamber with hinged Plexiglas lids. One compartment was 000542 418-008:PAGE II-16 (oo1uuttmffAitt)eeddwwawiistthhdeaaligbvrreiirdgehfdtl.oloiTrghhttoeawtnhwdiocPhcloeamxipbgralireatfsm(e1flsoeorc.onTdh)epuoltsheerocfommilpdaretlemcetrnitc wcausment nts were separated by a sliding door. dtDhoueoriranpwgpaaesraacothpuestnriueanld,titahintederntathteewraleisdghttphlweaac"seddatriukn"mteocdotmhopen.a"rbtrTimhgehent"ractowmapasratlmleonwte,dtthoe.selxipdlionrge Pdilemlamicevedediraeitdnettlooyatchhleoolsgderidin,dgtflchoeor.ighTthwearsattwuamsedthoeffn, raenmdotvheetd.brfirTeohfmeptushllesiedaiponpfgacdruoarotrruenswtaassnadgt.hen 50 0 soebeated seconds on age until the rat each of two rfoerm3a0inseedcoinntdhseb"ebfroigrhet"thceosmtpaartrtomfetnhet nceoxnttitnriuaol.usTlryiaflosr consecutive trials (the criterion for learning) or un] m1a5xtriimalus mwe6r0e-sceocmopnledtiendt.ervTahlewalsaterneccyortdoeedntfeorr etahechdatrrikal,compartment or the tDfhroeasmlabageterenogcfryot(uiipnaslsewLcoeortnehdescc)ormittopearerinoetnedrinotnthheteh"fdeiarfrsotkl"sleocswosiminpgoandre(topmveeenrnadtlelnfrltoemmaentaisnugrpeesr:fotrhmeance); tceonmdpeanrctymeinnta onnovterlialen1voifrotnhmeenfitr)st;tteshtesleastseinocny ((ianctsievictoynldesv)eltaonednhteeexr"pbltrohiregah`tt"odrayi s(cesohsmospriato-rntte(mrlemonnrtge-tfterenotrmimontr)he;teetn"htbeiroingnh)ut;m"abcneodrmptohaferttilamaltesennttcoyotn(hientrcisarelict2eorniindonthien tfihreststeesctonsedstseisotn (cloomnpga-tretrmmenrettefnrtioomn)t.he "bright" compartment on trial 1 in s) to enter the second the "dark" test session lgOeunaarlpnuioanstgtepdaanridntuammewmdaotaeryry-.7f0i,lEeoadnceMh-mrmaaalt zewearasftotraenosvdteerodtniecnofoarewdmaiatnleaerttirioagnth,ftrs1ow6mimemaicnhgliattbeir lw,ere osaftpep2er1loxmCiomd=iaft1ieelCdy).Mni-nOmeanzieen.cahesT,heanmdatzhee wwaatserfiwleadswimtohniwtaotreerdtofoar td-eegmpaptuehgreaoftsutraei(nrsasnge o(nbaesoefotfhethtewMo-gmoaazlse osftctehhmetfeMas-rtmtthaiezsaet,,ftrhionemorrtatdheewrattswoopbaleramcrsee)dmoiavnntedodtrhfeerquosimtraterdhteitnawgastpeworis,tmitoOonn rtheemofivresdrifarl,omthteherawtawtears. wicorrect goal during the rTeqhueirienidtitaol aenrtmerchbootshenaomnstroifalt1hewmaaszdeesbiegfnoarteebdetihne.g remaining trials. Rats that faile wcehoriecethweinthrienm6o0vesdecfornodms in any given the water. A trial were guided d to 15-second intertrial ttohemacokrereactcgororaelctangdoal interval separated Sreieaasclshsiotronaltw.earsmEia1nc5a.hterLatathteweatnescsytr(esqemsuesiairsoenud.rteodTrhieneacsmheacaxonicdrmisut)emrtioonnucmohfbefirveofcotnisaelscuitniavneyertreosrtiess. otfheermraorxsi(miuncmor6r0ec-tsetcuomnsdiinnttherevamlawzae)s druercionrgdeedacfhorteriaalc,hootsrieal,thaes cwoarrsectthegonaulmobrer 000543 418-008:PAGE I1-17 iDEnoatescrahvagrlea;tgtwrhaeosucpotsrerswetecetrdegtocwaiolcmeap.nadrTehtdeheftocerrsittthesreeisofsnoilwoleonwrsienwgethrdeeespseaenpmaeraftoredbobtyh ateostnes-eswseieonks. nerurmobrser(ionfcotrrriealcst ttoucmrsitienritohneomnatzhee)fifrosrt edaacyohfttreiasltoinngt;htehdfeeirnastvtedmraeayaogsfeutrneeussmt.bientgrhotohfe oltfahteeernnrcuoyrmsb(iefnorrseoefcaoctnihadlstsi)atltooocnrriettaehcrehiosntehocenocntodrhredeasctyecgooofnatdlesodtnianytgr;oiafal nt2edsottfihntegh;eltafhtieesntacdvea(ryoagfetensutminbge; r to reach the correct goal on trial 1 of day 2 of testing. y in seconds) MFaelmealreatsrawtserweereevaelvuaaltueadtefdorftohrethaegeagoef porfevpautgiianlalseppaatreantciyo,n,bebgeigninninnigngononDL 28 DL34 atOasnbslieDgsL,nsewdi8t5thottcohoe9h8ae,bxicttlhauetsiFio'on1n,ogoefnnseeirbamltaiinlogenmrraaatttipsnegwrsi.tfheimTnaheleaeccrohahtd,abobisatasagteiedognornpoeurrpaionwddeocrmoenunit of a the maximum of 14 vaginal contents days. andlor Faecmoapluelartaotrsywiptlhugspoebrsemravteodzoian observed situ were sisted in a smear considered of twSioatbmhieenadttohDesaGfigre0st7garndoduaapysstshoiafgtncheoahdadtboimtaiatntedidiov.nidwuFaeelrmehaolausseisnrigag.tnsewdFeearmletaelarlenlaortawetesmdatthloaetnradaitds nfortommatthee. deliver and maintain litters through a 21-day postpartum period. turally Tgehsetsateiorants(DwGer0e evaluated to the day for the cflirisnticpaul pobwsaesrvoabtsieornvseddu)r,inligteprarstiuzreit(iaolnl,pduuprsation of sddteialetiduvsebreaeftdo)rbieratnhi.nditipTaulhpeexvalimaubinignlsaittywieaotrneboirfrthet.hmeoPvlueitpdesrastnfhdoartivemiiamtbehierlirsteyadpwpeiernaewraeetxdears.mtiilnPleubodrnfoorrvtithaalt clounnsgsidtehraetdsalinvkebwoerrneancdontsoidhearveed dsiteildlbsohronr;tplyupasftewritbhirtlhu.ngEsatchhatlfitltoeartweapdsswewirteh wsuabsseeqvuaelnutatleydedxaaimlyinwehdendaitlhyefpourppsupwevriaebileixtya.miMnaetderdnuarlinbgehtahvei2o1r-odfatyhe dams wp1oh4seatnnpdaprr2te1.suemntVpe,arrioioandt.ialolnOsobtsfhreeorrmvdeeadxypsmeaoctfteetrhdneamlpaotbseetrhpnaaavrlitoburemhwpaaevsriioordre.cwoerrdeedreocnorDdeLds,1i,f 4a,n7d, TFehretsietyppaarraammeetteerrssiwncelruedeadsstheessfeerdtlftoyr ailnlddexam(spearscseingtnaegde toof mnaattuirnaglsdetlhiavtery. rPreuespsusul)ltt,eeddnuiinmn btpehreergbonifarntihcmipoelfsal)ni,tvaegteliisotttneartssii)t,oensn,uinmgdebenexerr(aoplfeocrfocfnesdnpitrtaiignogen pooeffrptrhlieetgtdenraa(nmlicviaeeasntdhadtead dsuurrviinvgetdh4e paonsdtp7adratyusm),pearnidod,lavcitaabtiilointyiinnddeixce(spe(rpceerncteangteagoef opfuppuspbsorbnnodrtnfhiattttherat survived 21 days) 000544 G9. Gross Necropsy": 418-008:PAGE II-18 G.9a Fo Generation Male Sample Collection and Female Rats Assigned to Pharmacokinstic Asraittrssedcahlleiltdotuewrlesedodftosdanacarmtiusfriacalelllyaofdwteeelrdicvtoeormnpaaltefuitrtitaeolrnl)oybfldeotlohidevescraoahmaplbliitettesar)t(iaaonpndpproeornxiioDmdaL(2m1al(efermaatslethat irnatt)o wseerreumcoslelpeactreadtofrrotmubfeisvearnadtscpenetrrdifousgaegd.e Tghreourpesfurlotmintghseeirnufemrit(oearlpvyper4noaxmicLmaavtpaee,lry 2shmiLp)mewnatstoimthmeedSipaotnelsyorfrfoozreannaolnysdirsy. iTceheanldivemraiwnatsaienxecdisferdo,zwenei(g7h0ed,C)anundtila sample section the Sponsor for (lateral lobe) analysis. was frozen and retained at ~70C until shipment to pTIeVhrethlailtitevrew,reofrfreoezsaeecnlheacnptudepdrfefotrraoipmnhetadhrematalci-to7ek0risnCoe,fttiunhcteislfaismvhepildpeamemcnosltlietncotetiahocnehwSoapfosnGesrxoocruipsfesodr,I tphorooluegdh analysis. surviving The pups dams in the on DL 21 3.2 mg/kg/day dosage group (Group V) did not have G.9.b. Fo and F1 Generation Male cvMioashlcaeebrirataatwtsaiwosnerppeeerrfsiaoocrdrm,iefadi.ncdedGabrgyorscosasrslbenoseinocndrsioopwxseiyrdeeofarstephtheayitxnhieoadrtaiicnoicnn,eauafttbredarlocmboiumnfpaflelertaeidnodn poeflvtihce gi1nr0doi%svsifdloueralsmliayolniwsneifaorgrehpeaodvsasiailnbadlbelrefeutitnauitrnheeedevrfaaolrwuapdtoaistosani..blTeRhehepirsfteoosllelonogtwiiacntegivvoaelrugpaahtnoisotnow:gerrtaeephesxcoifsed, weepirdeidfyixmeiddeisn,Bopurions'tsatseolauntdiosnefmoirn4a8l vtoes9i6clheosu(rwsitahnadndthweinthroeuttaifnleuidd).in stes, The testes neutral obrufgfaenrsedwe1r0e%rfeotramianleidninfonrepuotsraslibblueffheirsetdop1at0h%olofogrimcaalline.valuation. The remaining G9. Fo Generation Female Rats Assigned to Caesarean-Sectioning rUFtoeemsriaslonefearcparptosaprasesynstoilfgyntnehodenttpohroeCragacenisacan,rtearabandt-ossmewicentraielonsaitnnadignpweedelvrwieictshvaic1src0ief%riacaewdmamosonnpDerGfo1r0m,ead.nd a confirm the absence of implantation sites. All ovaries and gross ium sulfide lesions were to a AFt1agbelneeroaftriaonndroatmfurnoimtsewaachs suesxedfrtoomsewlheiccthoanlel tciosnstureosl egxraomuipnFeodaantd nhiesctrooppasthyowloegriecarleteavianleuda,tiinonosrodfergrtoospsrolveisdioensc.ontrol tissues for possible 000545 418-008:PAGE II-19 Rfraeettpsariweneseerdnetinaetxniaevumetirnpaehlodbtufofogfrretarhpeedhsn1uo0mf%bgerfroosramsnalldeisndiifosontrrsipbaorsesiabvlaeilfaubtluereinetvhaleuraatwi.odnata The omvabrryyoanwdaismpolvaanltoarticornessicteenst, and viable and ution of nonviable corpora lutea embryos. A in cach visgs shaped, pink, fim and enclosed in an arnigticsac cfflialoneudodryw,dieohrecpolperaaerqduftleoidbf.lluaidcA.k,nEosnmofvbtiraaybnoldse eewnemcrbleroysdoeidswciaansradanemdaomarfnpihootuisc,sascmaflill,edpawliethpiclnekart,o ter examination. G94. GEeonGeernaetriaotniFoenmFaelmeaRlaetsR.ats Assigned to Natural Delivery and F1 AwtertehesaccormipfliecetdioonnofDLth2e1,21a-nddaya pgorsotspsanretcurmoppesryioodf, pelvic viscera was performed. The number and ail the dthaomrascitch,atadbedloimvienraedl liters and was liter recorded. Female were sacrificed on rats DG assigned to natural ddeilsitvreirbyuttihoantodfidimnpoltandtealtiivoenr ites 4 25 and examined for gross lesions. To confirm the lp1ars0et%gpnauapmnmcwoyanssitfautomuusn,sduuldtfeeirdaied"f..rmoimDsarsamitnssgtwohiratthparnpeopsesuaumrreveiddvicnnagonnnppiubrpaelsginwaenrtewsearcerifsitcaeidneadftweirtthhe roefttahienetdhoirnacniecu,traabldboumfifnearledan1d0%peflovricmavliisncfeorra pwoasssipbelreffourtmuzereded..evAAallugaortvoiasorsnni.eescrwoerpesy uwRtaaetsrsimntaehdaceto.ndtieeTndhtwesewrraeetrseewxreaermceiornedexedda.mfiornOetvhdaerficoerasugsrweoesroseflrdeeestiaaotnihsn.eodnPitnrheegdnaaynctyhestoabtsuesrvaantdion formalin, neutral buffered 10% G.9.e. F1/F2 Generation Pups PwOeunrpesDLtwhe2er1,esasFma1cergieafnsiectrehadotaisnoednuepsxueapdsmfionnroetpducpofsonrtcigunrluolesedds oolennsisDotnLus.d4y Naencdraolpl sFy2pgreonceerdautrieosn gvPirutoaplssssttlahetasutisodniastedabinbrdtehf,foorarestehdxeeascmcairunisabteeidoopfnrdeoevfaitothuh.selylP.ituePprusfpowrsiptfhuopugnrvdiadbieliatdy wweerree eevxaalmuianteedd ffoorr DDLLss161to0421wewreereprperseesrevrevdediniBnonueiunt'rsalsobluuftfieonr.edG1r0o%ssfolreomssailsoinnls.esfirRooenmpsrpfeuospuensndtfaootuninvde on photographs of pup gross lesions are available in the raw data. sdPiiunopgxlsiednecoratossspseh-lysexecictatetidioonfnoroafcntodhneteixhnaeumaeiddnaeetvdatlfhuoeartlgierovoenslosonfletDshLieon4fsr.wonetrNeecsraocprisfyiciendcblyudcearabon examination of the cross-sectioned head for apparent al and parietal hydrocephaly. suture The and 000546 418-008:PAGE 1-20 tsahtneodsmelallct. hhrSecaeomndptolesenatssgwe(emgrirleokuccpouslr.lde)cItwneeddirvfeirdocumoallallpecuptpuedpssafrmfoprmloemcsufliwlveeedroepfuctphosembfliarrnogemedsGtrbloyiutitpetrsesrIi,niInto Polypropylene samples were sthuibpepseadn(dfrforzoeznenonatd2ry0icCe.) After to the Scpomopnlseotriofnoroafnsalaymspilse collection, 000547 G.10. Statistical Analyses: 418-008:PAGE I-21 The following schematic represents the statistical analyses of the data: Type of Test I. Parametric A. Bartlett's Test II. Nonparametric* A. Krus(ka5l7-5Wa%ltliiess)Test Significant atps0.05 Nonparametric Not Significant | Analysis of Variance Significant at p<0.05 Dunn's Test Not Significant Significant at p<0.05 | Dunnett's Test Not Significant B. Fisher's Exact Test (>75% ties) II. Test for ProportionData Variance Test for Homogeneity of the Binomial Distribution a. b. UStsaetidstoincallylytosiagnnailfiyczaentdpartoabawbiitlhithioesmoagreenreeiptoyrtoefdvaarsieaintchee.r P<0.05 or p<0.01. . d. Proportion data are not included in Test for homogeneity of variance. this category. 000548 418-008:PAGE II-22 BPirnoopmoritailonDidsattraiwbeurtieTMoan,nalyzed using the Variance Test for Homogeneity of the cCoonnstuimnputoiuosndadtaata()e.wge.,reboadnyalwyeziegdhtuss,ibngodByarwteliegthtt Variances"? and the AnalysisofVariance", wh TcehsatnogfeHsoamnodgefneeedity of Test was (p<0.05), nDoutnsniegntitficTaenstt(">?0w.0a5s))u.seIdftthoeiAdennatliyfsyiteshneofsatVpaaptrriositpairncicaaletsewiagfnsii.fsiicBgananicrfeotao'nftsthe. r0inedscitavsiwdeausaslwsghiregonruiepfsit.chaentIKf(rtpuh<se0kA.an0la5-l)Wy,aslitlhsseoTfKerVsuastrkiwaala-nsWcaesltlwatiaissstTinecoasltltyaTMpspiwrgaonpsriuasteed[i.e(<7B5a%rieotst)e sDiugnnni'fiscaMnectehoofdtohfe MiunldtiivpildeuaClogmrpoaurpis.sonIfst"h?erweawseruesegdretaotiedrenttihifafinycat7nht5e%(spt<ta0ti.ies0,t5i)o.a! eFvisahleura'tseEnxeaccrtopTseystda"twaafsorutsheed.pupFsiswhehri'cshEwxeacrte Tsteilslbtornwoarsfaolusnodudseeaddtq drDieasftclaerxeoitbpehtdyasaiitncaeadl(ead.tge.vC,eanleuosapmrmbeeeanrnt-aoslfeccdtoairtopanoairnangdl,uptnoeaast,tuwrneaulamndbieelnirgveobrfeyh,pavpiroerwaeladnaitnag involving criterion). were evaluated by the Kruskal-Wallis Test", asupdsespcerrilbietder.abtoavles,fo a 000549 Wl. RESULTS - Fo GENERATION MALE RATS 418-008:PAGE Ill-1 A. TClaibnliecaBl10O)bservations (Summary - Table B1; Individual Data - No Fo generation male rats died during this study. oiAlnnlceicldoiernnitccwaelos orwbaetssre. revTanothtieosdneossowabegsreee-rdvcaeotpniesoninddseenrinetcdalnuudndrleedolrlaottcheaedliotzboesdtehraevlatotesitonarotciccluerbreedcaiun soenltyhe acbormaesailonopoancitthye, ndeecnkt,alhyppreorbaclteimvsit(ym,iessxicnegs,sbsraolkiveantioorn,micsharloimpgeoncdeiadacrionyncoirsttohrhese)ls,i,mbs. swollen snout, chromorhinorrhea and dyspnea, B. Body Weights Tables B2 and and B3; IBnodidvyidWuealigDhattCah-aTnagbelse (Figure B11) 1; Summaries - PrGser0do.uu0cp1es)d aibndomtdihnyeiw0se.ti4egrmhegtd/g0ka.gi4/ndmsa.gy/`kdTgoh/esdavagayeluagenrsdowuhepirgoehnesriddganoyissfiac5ga6entstlooyf6r3tehdoeufctseesdt (aprt<i0c.l0e5hoard 21.0263m)g;/ikngtlhdeay1.d6omsga/gkeg/gdraoyupdoonsaDgSesgr1o5u1p0 on DSs 29 to 22, 29 to 36, 36; 36 and to 42 tudy (DSs in the. and 56 to 56 63. arRneefddlue3cce2tdinm(ggplt<kh0ge./s0d1e)aeyffofdreocttshsaegoefenttghirreoeutppesrs.tecaorThtihacbeliet,0a.b4t,oido1ny.6pweearinigdohdt3.(g2DaSimnsgs/1kwteor4e2s)iginnitfihcean1t.l6y egnrtoiurpessthuaddy,sicganlicfuilcaatnteldyfrreodmucDeSds(10<t0o.6035 oorr fp<r0o.m0D1)Sb1otdoytweerimiggnh/attdigaoayni,dnsosfaorgethe hgBraoodduyspiwgoenniifgiDhcStasnstwl5ye6,rree6d3suicagneniddfi(capat<nst0tl.yu0d1ry)edtbeuorcdmeiydnaw(teipio<gn0h..t0s5T)ohneinD3t.hS2e m1o.6/kmgg//dkagy/ddaoysadgoesaggreoup study termination. 35, 42, 56, 63 and at Body weights dosage of the and test body weight article. changes were unaffected by the 0.1 mg/kg/day C- Absolute(g/day) and (Summaries - Tables BR4elaatnidveBS(:glIknadlidvaiyd)uaFleeDdatCao-nTsaubmlpetBi1o2n) Values aRenldat3i.v2emfge/ekdgc/odnasyudmopstaigoen gvralouuepss woenrDeSssign8iftioca1n5.tlyArbesdoulcuetde relative (glkglday) feed consumption values were significan (p<0.05) in the (g/day) and 1.6 rinestuhlet o3f.2thmegs/ekgef/fdeacytsdoofstahgeetgesrtouarptioclne,DaSbsso1l5utteo a22n,d2r9eltaotit3vle6yfareneedddu3cc6oendtso(u4<m20p,t.i0Ao1sn)a values were reduced and/or significantly reduced in these two dosage groups for 000550 418-008:PAGE lI-2 t((hpDe<S0se.n0t55i6roe1r0ppr6<e30)c.,o0ha1ab)bsioitnlautttiheoenf0ep.ee4draicnooddn(s1D.uS6mspmtg1i/otkong/4v2da)al.yueAdsfotwseearrgteehesgicgonhiafibciatnattliyornepdeurcieodd relative dosage fgereodupcodnusriunmgptthiiosnpverailoude.s were significantly reducerdouipns.theAb3.s2olmugthekgalndday Absolute and dosageofthe relative feed test article. consumption were unaffected by the 0.1 mg/kg/day D. Mating and Table B13) Fertility (Summary - Table BS; Individual Data - a(Dnnoudsmafbgeeretrisiotofyfpptraheregantmeaesnttceairrestsieclpveaelrausnatuhemidgb.heraVsaofl3u.re2astmsgfio/nrkctgoh/hedaafbeyirttdialitidtiyonnaontadnafpfrecetgnaannycmyaitnidnigces arensdpetchteivneluym)b.etrheofnuramtbsewritohfcdoanyfsirtmoeidnsmeamtiinnagted,attehse dnuurminbgerthoedffrriarastttsswttehhaaettkmmoaaftteedd. cohabitation were comparable among the five dosage groups. E. Necropsy Table B14) Observations (Summary - Table BY: Individual Data - deAfofsseiacgtgnsiefoigfcraPnoFtulOpyS.ihnacdrTaehaelsieggdhrto(sbpsr<o0l.we0ns1i)oornnsbuormfobwtehnrelolivfievrme,raawlneerroeabtscseorinnvsatithdieeorn3e.da2ttrmre/ikbugtieddatoy test article because the incidences were dosage-dependent. lated to the gtOhrneoesuep1.mr6aatlmseg(kr8ag2t4/s5d,maay8t2ed5do2,sbau8gt2e5d7igd)ronhuoaptdirsammtpar(l8el,2g1nf4al)taceacniaddfatenhmdraleloeer3rp.aut2.rpmlge/ktegs/tdesa'y. dosage Al four of dfeomsaalgeeragtr. ouTphemaglreosrsatl(e8s2i4o3ns) ohbasderavsemdaliln tphroesttaetset;esthainsdraptroiOsmntpaertee3g.wn2eatrmeegd/nkaogt/day ocatothnsesraidnmedarledeiddrtanetosstthavaradtrcylgerf-orrsoesmlalHteiessdtiooarnsisctaohlfeCtyohnaetrrreoeclproiomndcumicodtneinvecfieonardtgiantnghsse.iTnecsotnitnrgolFagcriloiutyp. maNloe Abilnelccioatduhesenerc:ense1wc)ertorhpeesnoyobotsbedsroevsraavtgaieto-indoonecspcewunerdrreeendtc.ionnsoTinhldeyesroeendeobursnaertrevilnaattaeidgortooupt;heorte2s)t tahret.icle liver, a liver with a rough surface, constricted papillary procensssionfcltuhdeedlivaerlwarigfe a FIanc2il0i7ty9fcroonmtrAoulggursotu,p1r9a9ts2 ftrooAmpr9i1, s1t9u9d9i,esthceorneduwcetreed2a1t rtahtes Tweistthisnmgall flaccid and/or purple testes. 0V0551 418-008:PAGE III-3 pinpoint cysts in raised tan areas, the parenchyma, opaque lens dilation of the of the renal eye, large pelvis and kaidlnaerygse,spkliedenneys with F. WTeerimgihntaltoBToedrymiWneailghBtosd,yOWregiagnhtWe(iSguhmtmsarainedsRa- tTiaobsle(s%)88ofaOnrdgaBSn; Individual Data -Table B15) PsTseh0me.i0n11a.,l6 rvaeenssdpiec3cl.te2isvmewgliyt/)hkgtf/leudrimadiyanandldosbtaohgdeeyprgwoersiotguahpttses.hwaeTdrheseisgiangbinsfioifcliaucnattneltylwreeirgdehutcsedof(t5h0e.05 and Ep0i.d0i1d)ymiindtehsea3n.d2 mtegsltkegslwdeaiyghdtossaagned group, compared to y the condturcoeldgr(o2up0..05 weights of the seminal vesicles without or fluid were unaffected by dosages of the test article as high as 3.2 ma/kgiday. sfToihgtenhiferiacctaoinnottslryooflitnghcreroewuaepsievgdahlt(ue5os0f..t0hT1eh)leefistneathonebds3re.ir2gvhamttgtieo/sntsdeswaetyoretdeoarssmasiongcaeliagbtrooduypw,iwaesigchotsmpwaerreed sriagtniiofsiocafntthley reduced weights o(fp<th0e.0e1p)idtierdmyimniadlesb,odsyemwieniaglhtvsesiincltehsisadnodspaerdgoestgtarhtoeutpwh.eerTehe generally comparable among the five dosage groups. 000552 IV. RESULTS - Fo GENERATION FEMALE RATS 418-008:PAGE IV-1 A. CTlaibnlicealC2O2b)servations (Summary - Table C1; Individual Data - Al. Mortality No 1.6 dmeaahtkhgsldwaeyredoasttargiebugtarbolueptoratP,FaOnS.eveTnhteatotnrliybudteaabtlhe occurred in a to an intubation accident, esFbtesumedarylvdeaatriyaot(nD8sS9i)3n8c3l6iundatefthdeerc1ha.rd6moimmnogid/saktcrgraitydioaorynrdhooefsa3a5agndedaiglaryocdulopesawgaess.morAidbvuenrdsesaechrniifciacled on ofOtobhs3ee6rr,vtaianstsdiuoepnstsoasoipfsp,cehharuonmcohdeadcrpyoasrtruhreeaaannddlaptboosriesd wberreaeatrchioonnrgfailorenmxeuDddSaatt36en,eocnroDposyg; 3a5ll {rheidsurcatedwafseemdicsoinntsuubrmaeptdteidnoonornmoanolrDaatbSonue2tc9rDotSpos3y365... TTAhlhilseotcralhtienrliocFsatlobogobesdneyerrwvaeatitiigoohnntsfaeinnddihcaatded survived to scheduled sacrifice. male rate A2. Clinical Observation: TgOehbsseteairtnvicaoitndieaonnncsdeolofafcltloaoctcaialolinizzepededraialolodopspeecicniiatahwaeetr0a.e4n,yin1ac.rr6eeaaanswedads3d.u2srimginngig/ktgh/edparyecdoohsabaigteatgiroonu,ps. g(eps<t0a.t0i5onorpepr<i0o.d0.1)Thinetihnec0i.d4e,nc1e.6oafnldoc3a.li2zmedg/aklgo/pedcaiyadoonsatfghiceeanbgtarlcoykuipnwscardseusrsienqg the lSaicgtnaitfiiocanntpleyriiondc.reased (p0.01) in the 3.2 mg/kg/day dosage group during the eAplerlreioothdnesortwcdeiorisneiaccgaoeln-osdbiesdpeeerrvneaddteinuotnnr;sealdanutdre/idonrgto2t)htehtehperteeoscbtosheaarrbtviictlaetiboenc,augseset:ati1)onthaendinlcaicdteantcieons fTbarhboeoksreeendoobbrrseemaritvshaatliiognnsedininccliusdoresd).chrColemaordaocrarlyoerxutdhaetaea,atnipdtoondseiosnc,tcauhlrurpnercdohbeilnedmopsn.ol(symtiousnrseeinargac,t.s 1.6 mg/kg/day ing rat occurred that was mdourriinbguntdhesapcrreicfoicheadb,itaastipornevpieoruisoldy in the described, B. TBaobdlyesWeCi2ghthtrsoaungdh BC7o;dyInWdieviigdhutalCDhaatnag-eTsa(bFliegsurCe232;tShurmomuagrhiGe2s5)- B.1. Precohabitation sGirgoniufpiscanatdlmyirneisdtuecreedd(1p.<60.m0g1/)kgb/oddayywaenidghhtiggahienrsdfoosratgheesenotfitreheprteesctonaarbtiictlaethiaand 000553 418-008:PAGE IV-2 3p{.ep2r<0im.og0df1(k)OgiSlndstah1yetdo1o.4s62a)m.ggelDkgugrr/oidunapgyotdhniosDspaSegsreio8gdt,roovu1a5pl,uoe2ns2DwtSeosr2e92s9igtnoif3i6c;anatnlyd riendtuhcaed (cposm0p.a01r)edinwtihteh t3.h2e cmogn/tkrgo/ldgaryoudposvaalguee.grBouopdyonweDiSghsts15wa,en2rd2e,2s29i6g,tnoi3f3i66c,aannatsdly4r2e.dauqced compared with the control group values. Body weights unaffected by and body dosages owfetihgehttegsatianrstidclueriansg the high parsec0o.h4abmigt/aktgi/odnayp.eriod were B.2. Gestation tTahnhedepbr1oe.6cdoyahnawbdeiit3g.ah2ttimoggna/ipknegsr/iddoudar,yintdghoegsea1s.gt6aetsmigoo/nf.kPg/FRdeOafSlyedcctooinsntagigneeufefgedrcottuso trheadtucocecbuorrdeydwdeuirgihntgs tlheassn athned ctohnetr3o.l2gmrgo/ukpg/odnadyadyo0saogfegegsrtoautpiowne(iDghGe0d).siWgneiifgichatpntgtlaeyinnldeewsdasst(.o52w0e.i0g1h) asingdnitfihceanntelsysernetdiuaclleyd c(opm<p0a.0r5a)bline tthoeth1e.6comngt/rkogllgdraoyupdovsaalgueesgartoualpl throughout the remainder of gestation, beginning with DGs 10 to on DGs 0 intervals to 7 g1a0i7nsanwder1e0 stiogn1i2f,icaafnttelry wrheidcuhcethdisingtrhoeup3.a2lsmog/hkagd/wdeaiygdhotsgaagiensgcroomu12pp.aornWaebDilGgehsfto0the prceoerndtiurocodeld(gDr(Go<us0p..00t1Ro)e2fil0ne)ctathinendg3.bt2ohdmeysge/wkeegfi/fgedhcattyss,dwboeosrdaeygsewieggnirigofhiutcpagnfatoliryntsrheweeernetisrieggneifsitcaatnitolny 50.01) in 0 through the 20, 1.6 and 3.2 respectively. mg/kg/day dosage groups on DGsdu3cetdhr(opu<g0h.0151 or and dBoosdaygweesigohfttsheantedstbaordtyiclweeiagshthiggahianss d0u.r4imngg/gkegs/tdaatyi.on were unaffected by B.3. Lactation boTenheOreL3d.u21,cmeagds/ikcngo/tmhdpeaay0r.de4odsmwgai/gtkehggt/hrdeoauycpodnhotrasodalgsgeirggonirufopiucvpaanlotulney.DreLBdsoudcyedwe(i2g0h.t0g1a)ibnsodtyewnediegdhtto w3.e2igmhgtlklogslsda(py<d0o.0s1a)geocgcruorurepdwiansthpere1c.l6umdged/kfgr/odmayfurdtohsear1getevoag4l,ruoawtuhipoen(ntbhseeicgnaiufsiceanatll rDpeuLdpsusc1ed0id1e0dfit1bte4erfisonirzetehDeaLn1d.2)6.amsgsA/okcsigiga/ntdieafdiycwadinottshiarngecerdeugacrseoedu(pmpiw<l0ak.sd05ep)omsaisnibbloydyaswseoicgihattegdaiwnitohn nd and production. 0B.o1dmygw/ekigg/hdtasyadnodsabgoedyofwetihgehttesgtaiarntsicdleu.ring lactation were unaffected by the 000554 418-008:PAGE IV-3 . A(bSsuomlmuatrei(egs/d-aTy)abalensd CR8elatthirvoeug(ha/Ck1a/3d;ayI)ndFieveidduaClonDastuamp- tTiaoblneVsaCl2ue6s through C28) C1. Precohabitation ATinbhsetohsleeu1te.ef6f(eagcnt/dsdaw3ye.)2raemngds/igrknegil/faditciavayentd((og0/sk2ag0g/.ed0a5gy)roorfueppessd0.dc0uo1rn)isnuignmtpthhteeipo1rn.e6cvoamhlgau/bekistwateiroen rpeedriuocded group on DSs dosage group a2t2m0o.s2t9taabnudla2t9edtoin3t6er(vaalbss.oluRtefeloencltyi)n,g and in these the g/day dosage 3.2 mg/kg/day effects of the test aparebtrsiiocloledu,t(tehDaeSn3d.12rtemoloa4t/2i)kv.ge/fdeaeyd dcoosnasguempgtrioounp vhaalduessigfnoirfitchaentelnytirreeduprceecdoh(apb<i0t.a0t1i)on Feed consumption dosages of the test values article during the as high as precohabitation 0.4 mg/kg/day. period were unaffected by C2. Gestation Tanhde r1e.l6atainvde f3e.e2dmcgo/nksgu/mdpatyidoonsvaagleuegsroeaurplsycionntthiengueesdttaotihoanvpeerrieodd.uceAdbsaoblsuotleutaend arinenldtahtieinve1t.hf6eeem3dg./2ckomgng/s/dukamgyp/dtdoiaosynadgvoeaslagurgeoesugwpreoornuepDsoiGgnsniDf0Gictsaont70l1y(0arb7es,dou7lcutetoed1o(0np,l<y10)0.a0tn5odo1r72pt<o0.100,1) v((aaablbssuooellsuutoteecoaounnrldyr)e.rdeliSanitigtvnheie)fia1c.na6ndtalDnydGisn3c.1r28ematgsoe/dk2g0(/p(dsrae0yl.a0dt5iovsoeraogpnl:ey0)g.,r0oo1ub)spfesreoveadntciDooGnnsssui1mn5ptettrioo1n8 with reduced consumption body tends weights and to increase. the later Despite stages of gestation, when related maternal feed the increases in relative feed creodnuscuemdpt(ipo=n0.v0a1l)ueasb,sotlhuet3e.2femegdickogn/sduamypdtoisoangvealgureoufoprhtahde eantsiirgenigfeicsatnattliyon period (DGs 0 to 20) tFeesetdarctiocnlseuamspthiigohn avsal0u.e4smdgu/rkign/gdagye.station were unaffected by dosages of the C3. Lactation Athbeso0l.u4tmega/nkdg/rdealaytidvoesmaagteergnraolupfeaenddcwonesruemspitginoifnicvaanltulyesretdeuncdeedd (tpo<b0e.0r1e)diunctehdein 16 mg/kgday most intervals dosage group for within this period. tThheeen3t.i2remlga/cktgatiidoanypdeorisoadg(eDgLrsou1 ptowa14s), and at precluded from evaluation because there were no surviving pups after DL 1 000555 418-008:PAGE IV4 dFoeseadgceoonfsutmhepttiesotnavrtailcluee.s during lactation were unaffected by the 0.1 mglkg/day D. Estrous Cycling, Individual Data - Mating and Table C29) Fertility (Summary - Table C14; tcDhyoacstliawngeger(senoeufvmatblheuerattoeefsdt.esartAtrliulcslfeesatmsaaglheeisgrhapteasrsm2a38.t-2edadmyges/x)kcgien/pdttahoyend1ei5dinrnaottts apfefrecdtoessatgreougsroup dinodsiacgese(gnrouumpb.erTohfeprneugmnbaenrcioefsdpaeyrs ninumcbohearbiotfatriaotsn,thtahtemfaerthieeidt0y.aa4nndmdgr/paktrsge/gfdynaayncy ddcuoorhsaiabngigteatthgieroonfui,prsstr.easnpPderctesigevncealoynn)cd,ywaoencedckutrhorefecndouhimnabb3ie3tr,ato3i2fo,rna2tw8s,ewr2ie9thcaocnmodpnaf3ir0ramfbelde mianttihnegfdivaetes Groups | through V, respectively. emale ras in E. Necropsy Table C30) Observations (Summary - Table C15; Individual Data - kAtlihldenn0ee.yc4.ramognplsokybgso/ebdrsaveyartvdiaootsniaotgnhseatwgeirsroceupocmohmnasodindmeoirdneetdrhiaustnsreepdleiacltiaeetdisotnaonotdfrteshattermepenltv,isOonf ethreatriignht s1p:l6emegn./kgN/odaoythdeorsgargoessglreosuiponhsaodccaudrhreesdioinn tohfeafbedmoamlienarlatasdiinpatoihsni.es stOtiunsdseyu.eraatnidn fthhee F. TCaabelseaCr1e6a;n-SIendcitviiodnuianlgDaatnad -TLaitbtlereOsbsCe3r1vatthiroonusgh(SC3u3m)mary Cr7aee(ss7p0ae.cr0te%ia)vn.e d6soe(sc6ta0ig.oe0n%ig)nr,gouposb(.s3e0r.v0a%t)ioannsdo9n (D9G0.01%0)wperreegnbaansteddaomns10in(e1a0c0h%o)f, the five eTlimhtbeerrryeaovswe.erraeNgeonsodfbaoirmoclohogaripdcoarllilatyelriumtienpaow,rhtiiamcnphtlaotnrhteasrtteiatowinsest,ricevalinlaoyblsleiigveneimefbircaynotsdiofrfneornevncieasblian the mbryos. . NaantdurCa1l8;DeIlnidvievriyduaalndDatLiatte-rTOabbsleersvaCt3i4ontshr(oSuugmhmCa3r7i)es - Tables C17 sAnaalcttruhirofaiulcgedhdel2di5uverpriyrn,gesbtuehmeceapdursepemraeotgninenagnratpter(ra1it.os6dimn(goe/nakcDgh/Sdda3o6ys),daogasesagpgrreougprowuepr)ewaasssimgonreidbutnod tdoos25agweergreopurpeghnaadntonalnyd24derlaitvseraesdsilgitneersd. to natural deliveervyi.ouOslfytdheesscerirbatesd,. this 20 000556 418-008:PAGE IV-5 dnTouhsmeabdgeuerragotrfiooiunmpp,olfaangnteasottbiasotenirosvniattweiasosnpesarisgdsnoiacfmiicaawtnaetdslywsriietghdniupfcreedim(ppl<a0n.t0a1t)ioinn ltohses3[.2i.heamgvlekrga/gdeay resulting article. in a significantly reduced (p<0.01), icantly litter size] arnedduacnedef(fepc<t0.o0f1)t.he test P3.u2pmvgi/abkigl/itdyawyadsossiaggneifigcraonutplsy,readsudceesdcr(ipb<e0d.0i5n otrhep<f0o.ll0o1w)inign fthough the gestation index (the percentage of preg the 1.6 and information. Lhiaads icnocmrpeaarsaebdlneuamcbreorsss oalflpfuivpesdtohsaatgdeiegdroorupwse,rtehepr1e.ns6aunmmtge/rdkatgcs/adwniantyihbdalolvisezaeogdfefosgpnrrionugp) gpDrrLoeus1p.umahneaddd scpioagsnntniiifmbicpaallnaitnzlteyaditnoicnornDelaLosssesd2e1(v0pi4<d0e.nat0n5daso5rrpte<od07u..c0e1T)hpenuu3m.b2emrgs/kogfipduapysddoesaadgeor sliigtneirfhicaadntnloy ilnivcerbeoarsnedpu(pps<,0.t0h1e),nuthmebelirteorfadvaermasgweitadhndstniplulmbvoibranebirplutoypfsasttwibalibrsothrn[opnueps [wlsieivrgenebiofsriincganpnitufpincsuanmwtbeleyrreisnsci(rg0en<ia0fs.iec0da5n(tolpry<0pr.<e001d.)u0,c1ea)dnod(fpdt<h0ee.a0dl1i)ta)te,nr aasvewrealgleasanpderniupmarbteurmodfeaths nPuumpsbeornsD(Lps<0.10a1n)dof2dtaom4s(ahlladlivalelboprunpspudpesaddioerd pbrydeDspLurm2ee)sduamcnaedndnsiicgbananilnfiiizbceaadnltiozned (DpL<s0.10110)4]in. thReef1l.e6ctainndg 3t.h2esmeg/ekffge/cdtsa,ytdhoesvaiagbeilgitryouipnsd,extwheaslacstiagtniiofincainntdleyxrweadusced dawloessroaegstiehgengirafoivcueaprntablgeyegrsienfdnouircnsegudrov(nipv<Di0nL.g04p1)upopissntct(uhple<li01n..g06,5maognr/dkp<gi0n/.dt0a1y)dionstahgee1.g6romugp/,kags/daalyso group on DL 1 he 3.2 mg/kg/day dosage Agsirgdonouifspiacagadnemt-ildyneirpsetedenurdceeedndtth(pepa<tt0te.se0tr1na)rotpifculrpee. dbuoTcdheyed w1pe.ui6pghmbtgos/dkoygn/wdaelailygwhdetoigswaagseegvridoeunpt hiandeach w3e.2igmhgtloknglDdLay1d(onsoapguepgsrsouurpvihvaedd atosisgunbifsiecqauntelnytrsecdhuecdeudl(epd<h0wi.en0ig1gh)daipynsgu.sp).bTohdey The percentage of male pups was comparable across all five dosage groups. H. CNleicnricoaplsOybOsbesrevravtaitoinosnfsro(mSuBmirmtahriteosDa- yTa2b1lePsosCt1p9aratnudmCa2n1;d Individual Data - Tables C38 and Cas, gAardnovduepp.rosteeAndctvliiaenlricsraeeldaculncitdniiconanelciornbomspaesrtyveamotnbiasolenrcsvaaritneicoolncusdceuartsrhserodeceiian(tpte<hd0e.w03i1.t2h mlrige/dkugc/eddaypudposviaagbielity were been not nursing (two of removed, and one these litters had pups from which)thetptelrascweintthaphuapds that not had pups that were not nested). Apparent maternal 000557 418-008:PAGE IV-6 acanndniabaclainznaitbiaolniwzeadsfaolrseoliemvbidinenatnoitn htehirspduop)s,agwehigcrhoumpay(mihsasviengoctcaiulrirneodnaeftperup these pups died. Necropsy significant observations in pups increases (p<0.05 or that were stillborn or found p<0.01) in the numbersof dead included pups with no milk in itnhethseto0.m1acmhg/ikngt/hdeay1.d6oasnadge3.g2rmogu/pkhga/ddatyhidsoosbasegrevagtrioounp, pauspsc.omNpaorpeudpswit(hp<t0w.o01) control group pups. increased maternal cTanhneib3a.l2izmagt/ikogn/adtanyedcorsopasgye group aiso of stillborn had and evidence of found dead pups. Nine of these pups had missing hindlimbs and/or portion of the tail, cAlolnostihdeerrecdlinuincralelaantdednteoctrhoepstyesotbasretricvlaetbieocnasuisne:the1F)1thgeenienrcaitdieonncepsupwserweernoet odbosseargvea-tdieopnesnidnecnltu;deodr a2)dtohmeeodbsheeravda,tipoonrtoicocnuorfretdheintaoilnlmyisosnieng,littuemr.bilCilcianlichaelrnia, missing digits and anasarca (edema bent tail. One 1.6 of the entire body). mg/kg/day Necropsy dosage group stillborn pup on DL 21 revealed two had 0.4 mg/kg/day dosage lateral ventricles of the group brain, laintdteormnaetepsuwpitfhrommodaedriaftfeereonrts0e.v4ermeg/dkigla/tdiaony of the dosage group litte with slight dilation of the renal pelvis. I. RDeaftlae-xTaanbldesPhyCs3ic9atlhrDoeuvgehloCapdm)ent (Summary - Table C20: Individual Reversible delays in with body weight"? reflex and physical occurred in the 0.4 adnedve1l.o6pmmge/nktgt/hdaatyardeoshaigghelygrcoourpresl,ataesd described below. 11. Surface Righting Surface righting described below. was delayed in the 1.6 and 3.2 The percentageof pups that cmogu/lkdgs/udrafyacdeorsiagghtewgarsoups, as DsiLgsnif3i,c4an,t5l,y6r,e7d,u8cedan(d3100.,0a5nodripnst0h.e013).2inmgth/ekg1/.d6amygd/okgs/adgaeygdroosuapgoengDroLu1p, oanfter awthilceahstth5e0re%woefrtehenopuspursviivniangdopuspasgeingtrhoiuspdohsaadgtehegraobuilpi.ty The average day that to surface right was significantly increased (p<0.05) in pups ultimately attained the ability the 1.6 mg/kg/day to surface right. dosage group. All surviving The test ability article. to surface right was unaffected by the 0.4 mg/kg/day dosage of the 000558 I2. PinnaUnfolding 418-008:PAGE IV-7 (Pnionn3a.2unmfgo/lkdgi/ngdawyadsodsealgaeyegdroiunpthpeup0.s1,we0.r4e aenvdal1u.a6temdg/fkorg/thdiasypdaroasmaegteerg)r.ouTphse defofseactgewagsroturpasn,siaenntd (ceovnisdiendteroend oonflnyootnoexidcaoyl)ogiincatlheim0p.1oratnadnc0e.4[tmhge/pkegr/cdeanytages of pups P<0.01) per litter with pinna unfolding in the 0.1 and 0.4 mg/kg/day were significantly reduced (p<0.05 dosage groups on DL 3 only]. The or percentage of pups with an unfolded DLs 3, 4 and 5) in the 1.6 mg/kg/day pinna was significantly dosage group, and the reduced average (p<0.01) day that on at least 50% increased of the pups (p<0.01) in in a dosage group this dosage group. had All saurnviuvnifnogldpeudpspiunlntaimwaateslysihgandificantly unfolded pinnae. 13. Eve Opening Eye opening was delayed in 3.2 mglkg/day dosage group the 0.1, 0.4 pups were eavnadlu1a.t6edmgf/orkgt/hidsaypadraomseatgeer).groTuhpes (no defofseactgewagsroturpasn,siaenntd(ceovnisdiendteroend oonflnyoontoexidcaoyl)ogiincatlheim0p.1oratnadnc0e.4[tmheg/pkegr/cdeanytages of pups and 0.4 per litter with open mg/kg/day dosage eye lids groups were on DL significantly reduced (p<0.05) in the 0.1 14 only). The percentage of pups with datolseaagset ognroeuoppoenn DeLyse w14a,s1s5igannifdic1a6n.tlyThreedduacyedth(apt<0at.0l1e)asitn 5th0e%1o.f6 tmhge/kpgu/pdsahyad 1at.6lemags/tkogn/edaoypednoseaygeewgarosupssi.gnifAilclapntulpysinhcardeaospeedn(epy<e0l.i0d1s) in by the DL 0.4 21, and when pupil constriction was tested 14. Acoustic Startle PT<h0e.0t1i)meinofthdeev1e.6lompgm/ekngt/doafythdeosacaoguestgircouspta;rtnloe reflex was delayed (p<0.05 or 3.2 mg/kg/day dosage group Pups survived 1.6 mg/kg/day to be tested for this dosage group with reflex. The percentage of this reflex was significantly pups in the reduced (p<0.05 or ps0.01) on increase in DLs 13, 14, the average 16, 17, 18, day that at 19 and 20, least 50% resulting in of the pups a tendency for an had the acoustic startle reflex. 0T.h4emdge/vkegl/odpamyednotsoafgtehseoafcothuesttiecststaarrtitcllee.reflex was unaffected by the 0.1 and 15. Air Righting The ability groups; no to air right was 3.2 mg/kg/day delayed dosage (p<0.01) in group pups the 0.4 and survived to 1.6 mg/kg/day dosage be tested for this 000559 418-008:PAGE IV-8 0re.f4lemx.ghkTghledaefyfedcotswaagse tgrraonuspi,enotccaunrdrinnogtoonfltyoxoincoDlLogi1c6a.l Timhpeopretracnceentiangteheof pups (inpst0h.e011).6omngD/kLgs/d1a4ytdhroosuagghe2g1r,oruepsutlhtaitngaiirnraigshitgendifwicaasntsiignncirfeiacasnetl(y9<r0e.d0u1c)edin the. day that at least 50% of the pups could air right. 1.6. Pupil Constriction All the live pups in the 0 pupil constriction (Vehicle), response 0.1, 0.4 present and 1.6 mg/kgiday on DL 21 dosage groups had 000560 418-008:PAGE V-1 V. RESULTS - F1 GENERATION MALE AND FEMALE RATS A. F1Generation Male Rats A. MDoarttaal-iTtaybalnedD1C3l)inical Observations (Summary - Table D1; Individual A.1.a. Mortality gNrooudpeartahtss winertheefFo1ungdendeeraadtioonn dmaaylse8r6atasnwder8e7aptotsrtiwbuetaendintgo.PFOTSh.eseTdweoatchosntrol were attributed found dead on dtaoyin6t6ubpaotsitonweearnroirnsg.; One this d0e.a1tmhgw/aksg/cdoanysiddoesraegdeugnrroeluapterdat was. to the ftoersttharetsieclerabtsectahautsediietdwaarsae prsoivngildee,dnboenldoows.age-dependent effect. Observations Control and 11 mgirnouutpesmaalfeterratthe138170t5h was daily found dead on day dosage. No other 87 postweaning, adverse clinical one hour ocbosnesruvmapttiioonns ovaclcuuersrewderinetchiosmrpaat,raabnldeittsobootdheyrwreaitgshitn gtahiisnsdoasnadgfeegerdoup. Necropsy of the association with rat the revealed that all lung time death occurred, lisobceosmpwaetriebldearwkitrheda,nwihnitcuhbaitnion accident Choandtrboelegnraodumpinmiaslteerreatd 18351d1a6ilwyadsosfaoguensd.deAdadditoinondaalyad86veprossetwcleiannicianlg, after it o7b9speorsvtawteiaonnisngin).thisBoradtywweerieghltimgitaeidnstoacnhdrofemeodrhcionnosrurmhpetai(odnayvsal6u7estoin7t2hiasnrdat were comparable revealed dark red tcolotthtoedsemaotfeortihaelr(rpartes siun mtehids dosage group. to be blood) in Necropsy of the rat the thoracic cavity and a red accident. perinasal substance, observations compatible with an intubation Male rat 13138 (0.1 postweaning, after it mg/kg/day had been addomsiangisetgerroeudp6)5wdaasilfyoduonsdagdeesa.d on No day 66 other adverse ccloinniscuamlpotbisoenrvvaatliuoensswoecrceurcroedmpianrtahbisleratt,o atnhdosietsobfoodtyhewreriagthstignatihniss daonsdafgeeedgroup. All tissues appeared normal at necropsy. AA.b. Clinical Observations Ablelcaaduvseer:se1)cltihneicailncoibdseenrcveastiwoenrsewneortedcoosnasgied-edreepdenundreenlta;teadndto/otrhe2)tetshtearticle tohbeseirnvcaitdieonnceococfulrorceadliiznedonallyoopenceiaorotnwtoheratbsa. cAk significant increase (p<0.01) in the 0.1 mg/kg/day dosage in 000561 418-008:PAGE V-2 edoebpspeenndaeatnisto.nnooNctoccuoorntrsheiedrd.esrtAaedtddiistttriiceoanalatllymesanidtgvnreierflisacetaencdtliibnniecccarlaeuassseestihneavnalaudevewrasse neoftnidcoas|age. muinsraellaitgendedtointchiesotress)t, acrhtricolmeoirnhcilnuodrerdhedae,ntaablrpasrioobnleomnisgtn(hsmeioshbsesianedgr,,venbderaoarknetdnhecaonendyae/i.odrered tsfhnoeoruebtla,icmakb,orrleidbmapbcesrki,aonrcdah/lroosrumhboesdatadac,nrcyaeo,rsrcahaebraeo,dnslautchbeesrtabatanicockne, aoutnrtiphneenbisa,seloocfaltihzeedtaiall, oapescwioalloenn soft and or liquid feces, dilated pupils. swollen and purple ears, rales, gasep-isntga,ineexdcaebsdsosmailniaaltifourn, A2. Body Weights Tables D2 and and D3; IBnoddivyidWueailgDhattCah-aTnagbelse (Figure D14) 2; Summaries - wJTehoiewghp0t.o1pneardnadwye0e.1k4pmtoghs/atknwget/ahdneaiycnogndtoarsnoaldgggereongeurrpoaulrpaltsys,gtaewiintndheeddthoteoneewxeoirgthwloesgsrathmasnltehses cboondtyrol gwahiensn, tahsesceomgrpoaurpesd hwaitdhstighneicfiocnatnrtolregdruocutpiovnaslu(ep.<0R.e0f5leoccrtepipn<tg0i.ot0hn1is)ofpiandtabtyeosrdny7wo4efitogh36t, 9pwre7ei.cg6oh%htaobgfaiittnah,teiwcoeoningtphreotrligogadrion(usdpaiynva1tlhupeeo,0s.tr1weesapanendcitn0ig.ve4tlomy.gp/rkBegcoo/dhdyaabywietdaotsioang)ewgerroeup9s8.fo4r%thaend 04 the cmogn/tkrgol/dgaryoudposvaalguee,grreosuppesctfiovreldya.y 1 to termination ight gains were 98.5% in the 0.1 and and 96.6% of A3. A(bSsuomlmuatrei(easld-aTya)balneds DR4elaatnidveDS(;q/Ikgn/ddiavyi)duaFleeDdatCao-nTsaubmlpetDi1o5n Values taAhbnesdo0pl.<u10ta.e0n(1dg,l0dr.ae4ys)pmegfc/etkeigdve/cldyoa)nysadunomdpstraeigloeantgivvreaolu(upge/sskgdwiuedrraiyne)gsfiteghneeidffiitrcesatnnwtdleydrteodubceerde(dpu<c0e.d05in o(bdsaeyrsva1ttioon8spcosotmwmeoannilnyg)o,ccausr caonmdpwaerreedatsostohceiactoendtrwoiltghrtohueepetvkeanlodufeensi,nctyuTbfhaotreisoen 0sr.ie1gdnuimfcoiefcdkangbtlodidanycyrwdeeoaissgeahgt(epi<ng0r.t0oh5eu)speiontnwthdoeadyrosesla2at2gievteogf2reo8eudpposcsootnnsudmapyti1opnovsatlwueeaniinntgh,e A fouotnrhretehrliassttpeaodtirstttoiioctnahleolyftseisgtnairftiiccalnet bdeicffaeurseenctehseovcacluurerewdasin nwtoheteadnfoiesneagdgwce-aodnsespcuoemnnpsdtieidnoetnr.evdaNluoes the study. Ad. Sexual Maturation (Summary - Table DS; Individual Data-Table D16; ddDiaofysfoeafrgeenpsrceeposuftotichacelurtsereseptdaraarattmiicoolnengaisnththehiegthhFr1aesege0d.no4esrmaaggt/iekonggr/modauaplyse.diradtsn.otNaoffseicgtntihfiecaavnterage 000562 418-008:PAGE V-3 AS. DPaatsasi-vTeaAblveoiDd1a7)nce Performance (Summary - Table D7: individual tmTeahrlemererreawttese,nrteaiosnnoe,vlbaoilonulgao-tgtieecdramlblyyrepitemeprnotfirootrnamnoatrndcrieefsfipenroaennspceaessisniihnvibtihtaeiovnailnuetshefoFr1legaernneirnagt,isohnorttsrtiaatlisltaitceanlclyiessigonrifniucamnbtedrisffoefrernatcsesthoactcfuarirleedd tino tlheaernn.uembveoriodfatnrciaelspatroadcirigtme.rionN,o A.8. TWaabtleermD1a8z)e Performance (Summary - Table D8; Individual Data - rpNeeotrefbnitooirlomonag,inclcaoelnlgoy-ftitemhrpemorFrt1eatngetentndieoorsnaatogireorn-edmseapploeennsdreeatnistnhridebigiftafireodnri.enngcNeloseasortcnactiuingr,resdhoirnt.wtaetremr.maze. differences of rats that occurred in the failed to learn number of tials to criterion, trial lasttiecnaclileyssiogfnnifuimcbanetrs A7. Mating and Table D19) Fertility (Summary - Table D9; Individual Data - 2fDenordstiaftgeyretaisnitdoyfpptraheregatmneeastntecaryrstiiecnldveiaclaeussath(iengduhmianbsethr0e.o4Ff1mparge/egknnegardnaactyiieodsnidpmeanrlotenarfatfse.ct Vaanlyuemsatfionrgthe f{haetendumabnedrroaftsraitnsctohhaatbimtaattieodn,anredsptehcetinvuelmyb)e,rthoef rnautmsbweitrhocfodunamfyibstemrteodoifmnrasatetimsintnghaatte, J9doa6ts.ea0sg%ediugmrrpionrugepgstn.haetfOeifdrsttthwheeemecaokohloefrctroafthseambaaisltseaitgrinaotendwteorceohcaobmiptaatriaobnl,e8a2.m6o%n.g8t3h3e %thraened A. TNaebclreopD2s0y) Observations (Summary - Table D10; Individual Data - Adulenlrpeenlneadcterenodtp;stoyatnohdbe/soetrresv2ta)tairttohinecsloebinbseetcrhvaeauFts1ieo:gnecn1)oermtahmteoinionlncymidaoelcncecersatwsewreerenoctondsoisdaegree-d urs in this strain of rat, t(Ge1rs3ot1se3ss4,alen1sd3i1oen4ps5i,doif1dm3y1am4li8ed)e,rseepiarncotdhhurcoefteiwv(hepi<co0hr.g0sa1in)rse0d.i1nacmlliguted/rek).dg/fdTlaacycidd,ossamaglelgarnoduoprmapulreplreats Jcdoonssaigdee-rdeedpeunnrdeelnatt;edantdo t2h)ettheesstearftiincdlienbgescaaruesek:now1)ntthoehbeiesnecgieodnbeesntecirceavslalwtyeiormneesdniwoaetreed 2nd common in this significant incidence. rat strain. No other gross lesion occurred ata statistically 000563 418-008:PAGE V4 Aidndciltuidoendalrendecarbodposmyinoablseardviaptoisoensticsosnuseiwdietrhedfirumnraerleaatsed(otoneth0e.4temsgt/akrtgi/cdleay fdoolsloawgienggrionfuoprmmaatiloen.rat;On13e1602.1),mag/nkdgu/rdianyardyotsraacgtelegsrioounsp,maasledersactr(i1b3e1d43in)thhaed a rdielnaatledpeulrveitse.r;Oanneot0h.e4rmrga/tkign/tdhaisyddoossaaggeeggrroouupp (m1a3l1e34r)ath(a1d31s7li3g)hthdaidlaltaiorngeof the skliidgnhteydsilwaittihononfumtehreopueslvifsl,uiidn-fwilheidchcytshtesrien wtehreepasirxecnalcchulyim, aantdhetlheiftckkeindendeyurhiandary bladder walls observations associated with the in the control group rpartesstehnatcedioefdnausmethreoursesuclaltcuolfi.an Necropsy intubation error were described previously. A.9. WTeerimgihntaltoBToedrmyinWaeligBhotdsyanWdeiOgrhgtan(SWuemimgahrtisesan-dTaRbalteisosD(1%1) of Organ and D12; Individual Data - Table D21 wNeoigshtta)tisvtailcualelsy fsoirgntihfeicwaentigdhitfsfeorfentcheesroigchctuorrreldeftintetshties,absseomliuntaelovresreilcalteisve(w(ittohbaonddy rwietdhuocuetdfliunidt)h,eri0g.h1t aenpiddi0d.4ymmigs/okrg/pdroasytadteo.sagTeergmrionuapls;botdhye tweerimgihntasltbeonddyedwetiogbhets rinestpheect0i.v1elayn,dof0.t4hemgco/nktgr/odlagyroduopsavgaleueg.roups were 98.3% and 96.4%, B. F1Generation Female Rats B.1. Mortality and Data - Tables EC2l:inical Observations (Summary - Table E1: Individual B.1.a. Mortality No deaths in control group the rat F1 generation was sacrificed female on day r7atpsoswtewreeanaittnrgi,bubteedcatuo sPeFOitSw.asOanemale rat fthoautndhaddeabdeeonnidnacoyrr2e8ctploysitdweentainfiinegd;asnoa cfaeumsaeleofratd.eaOthnewacsontirdoenltigfrioedupforratthwisasrat, lOancteatriaotnin(DtLhe100.)4amsgt/hkeg/redsauyitdoofsaangeingtruobautpiownaasccfioduenndt.deOabdseornvadtaiyon1s0 foofr these rats are provided below. No gross lesions postweaning were revealed by necropsy of control group rat 13205 on day 7 pTohsetwceoantnrionlgg,raofutperfiet mhaalde breaten(1a3d2m2i1n)istthaetrewdas27fodauinldy dead on dosages day did 28 not have any other adverse clinical observations, and its body weight gains and feed 000564 418-008:PAGE V-5 tciosnssuuemspatpipoenavraelduensorwmearleactonmepcarropasbyl.e to other rats in this dosage group. All oFfemDaLle10,raatft1e3r2i5t 7ha(d0.b4emegn/kagd/mdinaiysdtoerseadgea gtortoaulpo)fw1a0s6 fdoaiulnydddoesaadgeosn. tNheomootrhneirng adverse t07. Its fcleiendicaclonosbsuemrpvtaitoinonvsaloucecsurwreerdeinuntrhiesmarratk,aballet.houTghhisit lost weight on DLs rat delivered 15 1 tlhievedbeoranthpuopfst,heofdwahmi.chNseecvreonpswyerofetchuelldead monreDvLea4leadndgreoisgshtlewseiroensscacormipfaitciedblaefter with light oarnainngteubfaltuiiodninactchiedtehnotra(cwihcitceavfiotya,mainndthdeartkrarcehdeal,unagpsp).roximately 2 mL of a B.1.b. Clinical Observations 0A.4remdg/pekrgi/vdaagiynadlossaugbestgarnocuepso,ccruersrpeecdtiivneolyn,e and two on days dams in 21 or 22 the 0.1 and of gestation, and red the perivaginal substance 0.4 mg/kg/day dosage was also observed on DL 1 group. These observations in five (p<0.01) probably were dams in associated awidtvherpsaertluyriatfifoenc,trtahtehehreatlhtahnoafntheeffdecatmosf otrhethteeisrtoafrftsipcrlie,ng.andNodidotnhoetraapdpveearrseto clinical observation occurred at a statistically significant incidence. gAlelstoatthieornaadnvderlsaectcaltiinoincapleorbisoedrsvwateiroenscotnhsaitdoecrceudrruendredluartiendgttohtehperteecsothaabrtiitcalteion, because: occurred 1) the in only incidences one or two rwaetsr;eannodt/odros3)agteh-edoebpseenrdveantti;on2)istcheomombsoenrviantitohne (lmaibsoarlaitgonreyde,nvmiirsosnimnegnta.nd/Tohrebsreokoebnsetrevetaht)i,oncshrionmcolduadecdrydoernrthaelap,rsowbollelmesn and purple ears, abrasion or localized alopecia scab on the back, on the back, urine-stained ahbeaddo,milniamlbsfuar,ndm/iosrsiunndgehrisinddep,aw digits, feces, perivaginal missing tail swelling, swollen sheath, tip of tail fbloarcekpaowr,mcihsrsoimnogrhainndoratbhreaas,iosnofotnotrhleiqcuhiedst. B.2. SMautmemranralieBsod- yTaWbeliegshEt2s tahnrdouBgohdyE7W;eIingdhivtidCuhaal nDgaetsa(-FTiagburlees4;E24 through E26) B.2.a. Precohabitation Body weights postweaning. tNeondsetdatitsotibceallryesdiugcneifdicianntthoer0d.o4smagg/ek-gd/edpaeynddeonstagdeifgfreroeunpceosnidnay 1 fpreemcaolheabriattsatdiuorninbgodthyewperiegchothagbaiitnastioornbpoedryiowdeights occurred in the F1 generation 000565 B.2.b. Gestation 418-008:PAGE V-6 uMnaatfefrencatledbobdyydwoesiagghetss aofndthbeotdeystwaeritgichlte gaasinhsigdhuraisng0.t4hmegg/eksgt/adtaiyo.n were comparable among the three dosage groups an period were All values d did not significantly differ B.2.c. Lactation w0S.ih4gincmihfgi/bckaogndt/yd(aw<ey0id.go0h5st)aggameiantgserrownueaprl,ebagosrdecyaotwmeepriagtrhhaetnd,lotosorstwoheecicgcuohrntrterldoolsosgnrleoDsuLspstvh1altuoe,4 in the after group values. an the control uMnaatfefrencatledbobdyytwheei0g.h1tsmga/nkdg/bdoadyy weight dosage goafitnhsedtuersitnagrttihcele.lactation period were 8:3. VMaaltueernsa(lSAubmsmoalruitees(-g/Tdaayb)leasnEd8Rtelhartoiuvgeh(Eal1k3a;ldaIynd)ivFideueadl CDoantasu-mption Tables E27 through E29) B.3.a. Precohabitation rrTeehddeuucc0te.i4donmrge(l/pak<t0gi.v/e0d5a()yg/idkngoa/sbdasagyo)elufotefeetd(hgec/dotaneyss)tumfareptetidciloecnownoasnsudmaapsytssiooc1inatatone8dd waitthenadseingcnyififcorant tohbissegrrvoatuiponosn that day w1 eproestawsesaonciinagt.ed with the Absolute tendency for and relative smallpeorstbwoedayniwnegi,ght feed consumption in vnoatluseisgnfiofrictahnetlryedmiaffienrdaemroonfgthteheprtehcroeheabdiotsaatigoen period groups. were comparable and did B.3.b. Gestation wAubneasrfoefleucctotemepdaanbrdyabrdleoleastaaigmveeosfnegoefdtthcheeontthserusetmepartdtioiocslnaegvaeaslguhreiosguhdpuasrsian0ng.d4tdmhige/gkegs/tdaatyi.on Aplelrviaolduweesre d not significantly differ B.3.c. Lactation (Tph<e0.a0b5s)olounteDLmsat1ertnoa4l fineetdheco0.n4summgp/tkigo/ndavyalduoeswaagse control group value. The relative feed consumption sgirgonuipf,icaanstlcyormepdaurceedd value was a to the shiogwniefviecarntnowteisgighntifliocsasnttlhyat(>o0c.c0u5r)r.edTihn etshies ogbrsoeurpvaotnioDnLsswe1 rteo 4asaslnosdcoiraaetttderudicbweuidtt,ehd tthoe the test article. 000566 418-008:PAGE V-7 Absolute and unaffected by relative the 0.1 feed consumption mg/kg/day dosage values of the during the test article. lactation period were. B.4. Sexual Maturation Table E30) (Summary - Table E14; Individual Data - dDaoysaogfevsaogifnatlhepatetsetnacrytiicnlethaes Fh1igghenaesr0a.t4iomngf/ekmga/ldeayradtsi.d not No affect the average significant differences occurred among the three dosage groups. B.S. Passive Avoidance Data -Table E31! Performance (Summary-Table E15: Individual There were no term retention, biologically important long-term retention or driefsfperoennsceesinihnibtihteiovnalinuetshefoFr1legaernneirnagt,isohnort- fsteamtiaslteicraaltlsy,siagsniefviaclanutatdeidffbeyrepnecersfoorcmcaunrcreedininatpheastsriiavlelaatveonicdiaesncoer pnaurmabdeigrms.ofNo rats that failed to learn. Trahtes innutmhbee0r.4ofmtgr/iaklgs/tdoacyridtoersiaongewagrsousipg,niafsiccanotmlyparreedducteodt(hpesc0o.n0t5r)olfogrrtohuepfveamluael.e Tihnicsreoabsseerinvatthieonnuwmabsernootfctorinaslsidteorcerditaernioenf,fercatthoefrtthheantesatdaerctircelaesbee,cwaouusledabne expected as an indication of toxicity. B.6. Watermaze Table E32) Performance (Summary - Table E16; Individual Data - pNeorfbioorlmoaginccaelloyfitmhpeorFt1angtendeorsaatgieo-ndfeepmeanldeernattsdirfefgearrednicnegs loecacrunrinrge,d sihnowrat-tteerrmmaze drietfefnetrieonnc,eslooncgc-utrerremdrientetnhteionnuomrberersopfontrsiealsinthoibcirtiitoenr.ionN,otrsatlatliasttiecnaclileyssiogrninfuimcabnetrs of rats that failed to learn, B.7. Mating and Table E33) Fertility (Summary - Table E17; Individual Data - aDnodsafgeretsitoyfptahreatmeesttearrsticelveaalusahtiegdhinasth0e.4F1mgg/eknge/rdaatyiodnidfenmoatlaeffreatcst. anVyalmuaetsinfgor the and npurmebgenranocfydianydsiciensc(ohnaubmibtaetrioonf,prtheegnnaunmcbieesr of rats that mated, per number of rats the that fertity mated amantdinrgatsdaitnecsohdaubriitnagtitohne, respectively), and the first and second week number of rats of cohabitation with confirmed were comparable among the three dosage groups. 000567 ~. 418-008:PAGE V-8 B.8. TNaebclreopEs34y)Observations (Summary - Table E18; Individual Data - uAlnlrenleactreodpstoytohbesetresvtataritoincsleinbetchaeusFe1:ge1n)ertahteioobnsferevmaatlieornastsocwceurrerecdonisnirdaetsreidn the Tcohnetsroelogbrsoeurpv;atainodns2)intchleuodbesderrovuatgihoanncdloomrmopintlteydoccocrutrexs oinf tthhies kstirdanienyosfwriatth, tcahlecsueliriantsthaelsKoidhnaedy and/or urinary a tan granular bladder material of in two the rceonnatlroclorgtreoxupanrdatst.hicOkneeneodf walls of dosage the urinary group dam bladder. that died aNsectrhoeprseysuoltbsoefravantiinotnusbaftoriotnheer0r.o4r mg/kg/day were described previously. B.9. aNantdurEa2l0;DeIlnidvievriyduaanldDaLittate-rTOabbsleersvaEt3i5otnhsr(oSuugmhmEa3r8ies - Tables E19 2Pr5efgenmaanlceyraotcscuarsrseidgniend22to (c9o5h.a6b%i)t,at2i1on(8in4.t0h%e)0a(nVdeh2ic4ie()9,60..01%)anofd t0h.e4 2m3g,/2k5g/adnady idnodseaxg(ethgeropueprsc,enretsapgeectoifveplry.egnAallntprreatgsnawnitthdlaivmesofdfeslpirvienrge)dwlaitstercs.ompTahreabgelsetation across the three dosage groups. vAisabiolcictyurirnetdhein0t.h4empgr/ekvgi/oduasygdenoesraagtieognr,outph.ereTwhaesnaumtbeenrdeonfcdyafmorsrweidtuhcsetdillpbuoprn pups DL 2 was also winecrreeaisnecdr,eaaslbeeditinnothtissidgnoisfaicgaentlgyr,ouapn.d the number of pup deaths after Aresdaulcseodoicnctuhrere0d.1inmtgh/ekgp/rdeaviyoduossgaegneeragtrioounp,opnupDLbo4d(yprwee-igahntds ptoesntdeueldlitnog)beand 0O.L4 7m,gb/uktg/bdyayDLdo1s4awgeergercooumpptaernadbeldettoobceonrterodlucveadl,uetsh.ouPguhpnbotodsyigwniefiigcahnttslyi,notnhe DLs 4 through 21, present on DLs 7 awnidth14s,ignaisficcoanmtparerdeudcttiootnhse(cposn0t.r0o5l garnodupp0v.a0lu1e,s.respectively) F2 generation pups body weights were comparable among all groups. aAddmvienrissetlryataifofenctofatnhye ottehsetrarptairclaemeattedroseavagleusataesdhaitghnaatsur0a.l4dmegli/vkegr/ydoarydduirdinngotthe l2i1t-erdasyizelsa,ctnatuimonbeprersioofd d(adumrsatwiiotnhoafllgepsutpatsiodny,inagvedurraignegs lfaocrtaitmiponl,anvtiaatbiilointsy and and live lactation indices, surviving pups per litter and pup sex ratios). 000568 8:10. CNleicnircoapl sOybOsbesrevravtaitoinosnfsr(oSmuBmimrathrito Day 21 418-008:PAGE Postpartum and V.g Individual Data - Tables E39 and es E40) - Tables 21 and 22; daNreotpiecclnleidneainscatlh;iogarhnndae/scorr0o.2p4)smytghoe/bkosgbe/srdevaravytaibtoeincosnawuoescerc:eura1rt)etrdtihbienutianbcliedteoncdeossawgeerse onfotdthoestaegste- rJcluaircneihicnaoglf.otbhhsaeedcrovdnaettcriorolenasasiendclmuodteodroancteivciotnytraonldgwraouspcpoluodpnlttyohtaathnewetaoosructnhow,toonpneaseptsi.pnagpToirhn c tail and one 0.4 mg/kngd/d0a.y4 dmogs/akgge/dgaryoudpospaugpe groups that had that an uhmabidliacamlishsainngafi of 0sN.po1emmciatllikkvigen/dsdatoyosmadagoceshaggorecaucgpusrr.oruepdOnipnuep2.coh4natdarnoadl gr4ariposuueppdsptautnhpatahawderheydfroaucnedpdhealayd ainndthoetnheree laterai lobes of the liver at necropsy on DL 21. rea on the median and Iek 000569 EFERENCES 418-008:PAGE V-10 T- d(Se1tt9ue9dc4yt).iDoenIsnoitfgentmoaxatisciioMtnyoadtliofCirocenapftrieoordnuecnotfc:ieonUo.nSfo.HramFromoondiasantdioDnr;ugGuAiddmeilniinsetroantion Register, September 22, 1994, Viol. 59, edicinal No. 183, products. Federal 2 RUeSg.ulFatoioodnsa;nFdinDarluRgulAed.min2i1strCaFtiRonP.artGo56o. d Laboratory Practice 3. NJPruaapmcabtnieceres2eS1t,MainMndiasartrcrdyhof2fo6r,HSea1af9l9et7th.y aSntdudWieelsfoanreDr(1u8g8s7,).MHGoWoOdLradbionarnacteory 4 OJEuEarCsoDpoendaentchiEesciaoocnncoiemrpietccaonCmcomemebnmyduanttihiteoynEu(or1n9o8pc9eo)ma.pnCEocuonnciolmidcecCisoimomnuonnit2y8Joufloyn {Leagbiosrlaattoiroyn.pra3c2ii(cNe.o.OfLf3ic1i5a;l 2J8ouOrcntaolbeorf):thel1-iE1au7nr.coepewaitnh Gpormiincmiupnlteisgosf.good MNCauhrttiiasogtneianalin,cTieMtc.yhSn.Tiescatansl.dIVnEofnyovtreimkra,toinPom.neEn.Stear(lv1i9Pc8re2o),t.eUcItnioVnivAogeRnecpyr,odWuacsthiivnegatnodn, D.C. Springfield, VA 22161 .S. Department of Commerce, 6. oCnfharlSitcsriteeixanoncn,eesM,(.SPN.roovc(1ee9em8d4bi)e.nrgsR7,eofp1r9No8ad3lu)t,crteJi.xvoCenlietno.SxyiPcmiptyosainudmt,erNateowloYgoyrekvAaclauadteiomnys of sychiat. 45(9) 7.10 7. LCL2oabnnotgrr,aotlPo.rDLi.aets(a,19iI8nn8ct.)h.eWCEihlmambrirnlygeotsRoani,nvdeMrFAeCtrail:DCeDv7e8lRopRmaetn.taClhaTrolxeicsitiyv(eTreratology) Argus Research Laboratories, Inc.) 01887-0630. (Data base providedby 8. IUnssetitoufteLaobfoLraabtoorraytAonriymAanlis.malNaRteisonoaulrcAecsad(1e9m96y).PreGsusi,dWeafsohritnhgetoCna,reD.aCn.d 9 iSmaplleawnstkait.ioEn.ss(t1e96l4l)e.n 247:367. aFmarUbteemruesthdoedreRaztutem. mAarkcrho.skPaotphiols.chEexnp.NaPcahwremisav,on 10. hlSoonwmeadoegScetonaret,ietGUynWio.vfetrahsneitdbyiCPnorocemhsirsaa,lnAd,imsWet.rsiG,b.utpip(o.1n9.6274)0S.tatVisatriicaalncMeettheostdsf,orsthEdition, -241. 000570 418-008:PAGE V-11 11. vSaorkiaaln,ceRsR.. BainodmeRtroyh,l,W.F.HJ.. pp. 370-371. F(1r9e69e)m.anBaarntldetCto'.s,teSsatnofFrhaonmcoigsecno,eity of 12. PSSptna.etid2se5t8ci-oc2arl7,5M.Ge.tWh.odasn,d6CthocEhdritaino,n,W.lGo.wa(S1t9a6t7e).UnAinvaelryssitiys PorfeVsasr,iaAnmcees., 13. sDeuvnenreatlt,trCeWat.me(n1t9s55w)i.th A a multiple control. cJ.omApmaerri.soSntatp.roAcsseodcu.re5f0o:r1c0o9m6p.a1r1i2n1g 14. WS.oHk.al,FrReRe.maanndanRodhlCfo,.,F.SJ.an(1F8r6a9n)c.isKcrou,skppa.l-3W8al8l-i3s88T,est Biometry, 15. DTeucnhnn,omO.eJt.ri(c19s646)(.3):M2u4l1t-i2p5le2.comparisons using rank sums. 16. Siegel, S. (1956). McGraw-Hill, New YNoornkp,arppa.me9t6r-1i0c4.Statistics for the Behavioral Sciences, 17. cLoorcrherlya,tiEonAo,f pHuopbebromdayn,weAiMgh.t landmarks. Teratology 29(2) and with Cohtrhiesrticaon,mmMo.Sn.ly(1u9s8e4)d. Positive developmental :44A 000571 APPENDIX A REPORT FIGURES 000572 | " | 7 5x3 : - TU g&a3 g Fo GEBNOEDRAYTFWIEOuNIeGMAHLTESraTS _ A 2 I Za 5 lx xox = FE - ==W O. VENCLE)MGKGIAY 0|| evmmonne on IT oToonaxconr = = 32MGKGAY a ae in aol i i 4 | - 0 Fo GENBEORADTYIOWNEFIeGwHaTLSe mars Figur2e i everson * i armawv <1 | concn et E:5o ii aoe"?7 x x { OR ;{ { svsexuancnr "| aE AT ES i [ps0.05 Eel lap EgAREET sg TEowoErstRuoySEIT ! yy BEB LRRE ERE 1) gag i a232 DAY OF GESTATION+ treOAe YOF LACTATION 3 8 z 1 F1 GEBNOERDAYTIWOENIMGAHLTESRATS Figur3e =| a=w= = || o=wes - frve on !o g &2 1 4 LE 3#ov En % wd dL ra, e g52 3 & mR. z7z F1 GENBEORADTYIOWNEFIEGMHATLSE RATS Figur4e | ] ml LrrrRsE5EuEaEasttt55 55 55070 gBge u ~24 5 ws | | ws | : "omc uaraon h 5 coma on : cone : ------ : <g Pb AaYnPOkSToWEuANnINaG s seOAhYObFiaGhEiSbTATIONkb ahgOAY| OFbLAChTATION 2 33 S-- 2 T APPENDIX B REPORT TABLES- Fo GENERATION MALE RATS 000577 | - I. FE SR PL a, eon nn so : i & & & 2 ] + EEE TE va ye pe i : 2 2 8 2 | ] mDeollOELIDDONMIDen n omem e s DRsIew i :$ 5 ? 5 i 7 i5 . i 8 i 2 " 3  So, pms. sur . . : ' g mm z re . ! : en. ai A SE Sys: . $ : 5 : g: 3 EI L MltL ot rn ss ion stn < 3 2 2a 23g CS Se REELS 18 rns ems io tesa g g z - ne 3: @ o APPENDIX C REPORT TABLES - Fo GENERATION FEMALE RATS 000587 ao cE i re i i rt LS 5 i 22 88 <2& Z i 2 2 BE ) cen JHE . : : i g -- te wil ern sn a are ERE ET mt seis a2 < g : g # 2 EEERI np er ) : g 2B3 & 8 g EE SHE Sn ee 2 8 g P+ EEERRETTHRRApNaEEm ESo 2 TUTTE i2 LER RRR 3 - 3 2g 5z2g 3i o EE EI nc sno 3 St hin dE reyae GIA er s 3 8 ag3g 2 3 EtTenet cre. 11 ton cies oh sin g -g2 32 Fg& 2 i 8 2 . Ls i = rv sin 2$i g: TM Tle ihe Tie? Tite he 7 5 1 BEI th SF ILShain nay 6 of aay mame fn el Str mins tee asian . 3 23 22 2 | ErE t 2 SER E, S tiRtnEos REson . g Sgg en 3 3 2@ 2 g2 & :2 g< 8 $g g . g 2 g 2 a $ 82 & g2 3 3& g2 82 823  38 22&82 2e BiH ME ER Ea omy . Ee EEEa Se Ree :i s&:g R E PEEER SElE EA RA anim ge ess ee vf  4 EERE Haar ace Z wJ oo. sos weeET we we oe a oe 2 voei om oma, me sors, sso wn ore wwe oo wr vous we oe .$ 23 & o 8 & s&:g T | EEHRe E Ei rhiE matm B-leeeee ee 8i# eo pos seting the cricaron fo th developmental ade Seted on each da ponparton 3 g.CeIHamirriaemsyySpioiI n tChiaat+eLrearAsi t SL a ou v 1s vknt pnopsnhad caphneeedelvelopmental sears presen. '8 &a 8 = : :: : Cg 28 =2Og 3 2 5 : : g z 0 wineserie or oopeciornd opus ln ichtolls or Chmiiaication prciude sviusion or g ERE 8 &ao a APPENDIX D REPORT TABLES - F1 GENERATION MALE RATS 600619 ae. eo mec we we ws iis war SOL Ree Sk Wh STISED 5 5 A o oS 4 SAO Ee IT JYLCs + a F meme teeny SBo G a LL i SI Te iSL oceon Soc weae.en : 3 2 ? = FRnmAa R Si Raa Cg 5g 88g2 8& 2 | of ERS LYSTes rc cnc vo oh 1 in se ion 3 EEE et evr at 8 g8 z | $ Em EERIAERE ERwE e rh src conmtnsoton te 1 io meme g & - 2 8g8 2& wenn mwas wi war ee meres S2 28 8] 8 i CPL EEL . i 3 3 Z 2 A gg - 3 2 : : g 3 % : Pa Refer to the individual clinical coasrvations able (able DI for di onerationbs a a mer - 2 gg8 i& mEiMmEimmonLm :? g igaiticantly different (rom the vehicle control group value (pso.ei) 3  i g 8 2& s 1 Ee VIE 20 TR Ih IIL ee wen aftectedt or orauns dasond eign affected -s 23 &8 o B. ELciied LLIAS (F Tits hat had sbooemst SrSane (velght affected) or organs dumped velght affected) o g 22 &g. z APPENDIX E REPORT TABLES -- F1 GENERATION FEMALE RATS 000634 PRCOUBITATIGN ton 1 oF $TVOY 0 THE ear oF CoumairiTion| gt ES 2% 33 B i i g tr tlLt CT ILsiAs a GI && . i3 7g a en ve wn 2 wm im g B E IERIEn m E i atAie a t a i ii azi g i QT Sun an tov cess onoa ot Sacatien : 2oF FRI Z2 8 STIL! ETE va cr ae ein cso 0 thn the 3 5 Tn g : PR g a LT IETTe wupen tor the #2 geacion sects 96 vias Sine shove i a ERR ETE mm on g< 2 3& fpr the texas essere faites 2 ' 3 -2 ]2 2g TT 2 Bienen gin fhe reg a pod peng Reboe 2 g28 82 " 2 2m8 EE TET a tr 22ios hs ty to 82 7J& : g<g zz 22 ILC ------ i g Z & EEERs nm $2 i ee eee inz  :{ g2g am2 83 Bo PI, sis mmm mip: est sesso <gsg3 &3Ti  HME DA SIS MS 3 g ig :&3? i $A MED ai a insrn stnpn he sn sy vii LE Me a : i &gg m3 $ ig a ?3 pr EEL DA BE AL PRS BLL, <ggg &8T: it a 222g && g 2 Rn32 ia i $2 gg ii kc iy i im sc ce fe i on oe tt ie 8: m& 4baoh tv ei Hee seis Ai Re SS seri io iiia cio 2 SE LI RI ei: he see eect nition 8 &ag 52 a ws or a inr 3 2g g &2 n 8G 8 FE oR BR nS ER AR g <gg &22 &4 he? 3 TL Miorvaes avers 3 fG CE E e IR urn ih ronsSeda E pn Seaton tyT1e0Aof hAceaon, tan pups ere sceiices 2m ] I&] Q2 LoBar' SET oBrpEosE TyEraRTIhNL ewoeTS se iRarAe Stru enLiea0l batons si pop ee cts - 33? R Z& 83 8 - 3a Sg m3 &g : g APPENDIX F PROTOCOL AND AMENDMENTS 000662 418-008:PAGE F-1 aeseancy Argus50R5esSehaerachhyLDarbiovrea,tBouriiledsi,nIgncA. K Aocnaronies Hors1ha)m,4P3enn7sylF.v2an1i5a815980474 -------------------------------------------------------------------------------- STUDY TITLE: PURPOSE: TESTING FACILITY: STUDY DIRECTOR: SPONSOR: PROTOCOL 418-008 SPONSOR'S STUDY NUMBER: 6295.9 CPeormibniatnaeld/POosrtanla(taGlavRaegper)odFuecrttiilotny,TDoexivceiltoypSmteundtyalofaPndFOS in Rats. dTihsetuprubrapnocsees orfestuhlitsinsgtufdryomisPtoFtOeSsttfroerattomxeinctefoffecCtrst/:COBR VAFIPlus male and female rats before cohabitation through mating, gestation and lactation. This study evaluates ICH Harmonised Tripartite Guideline stages A through Fofthe reproductive process and should detect effects on the estrous cycle, tubal transport, implantation. gestation, parturition, lactation and maternal behavior in female rats, on the developmentofthe offspring of the treated male and female rats, and permit detection of functional effects (e.g... effects on libido or epididymal sperm maturation) that may not be detected by histological `examinations of male rat reproductive organs. Because `manifestations of effects induced during this period may be delayed in the offspring, observations will be continued through production of F2 generation liters. Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Telephone: (215) 443-8710 Telefax: (215) 443-8587 Raymond G. York, Ph.., DABT Associate Director of Research 3M Toxicology Services 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 000663 ` 418-008:PAGE F-2 Protocol 418-008 Page? STUDYMONITOR: TMealrevpihnoTn.e:Case(6,12D).V7.3M3.,-5P1h8.D0. Telefax: (612) 7334773 SATLUTDEYRMNOANTIETOR: ATenlderpehwonMe.:Sea(6c1a2t), 5P7h.5D-.3161 Telefax: (612) 733-1773 REGULATORYCITATIONS: SInttuedrynaDteiosniaglnCaosnfMeordeifnicceatoinonHaofr:moUn.iSs.atFiooond; GauniddeDlriungeAodnmidneitsetcrtaitoinoonft(o1x9i9c4i)t.y to reproductionformedicinal products. FederalRegister, Septemb2e2r, 1994, Vol. 58, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. TJoarpSaanfeestey MSitnuidsitersy oonf HDeraulgtsh, aMnHd WWelOfradriena(n19c9e7)N.umGboeord21L,abMoarractohr2y6P,ra19c9t7i.ce Standard EacucreoppteaannceEcboyntohmeicEuCroompmeuanniEtcyo(n1o9m89i)c. CoCmoumnucniiltdyecoifsainonOoEnC2D8 dJuelcyis1i9o8n9/roencotmh-e tmheendEautrioopneaonn Ccoommmpulniiatniceesw:ithLepgriislnactiipolne.sof32go(oNod. lLa3bo1r5a;to2r8y pOrcatcotbiecer.): Of1f-i1c7i.al Journal of REGULATORY COMPLIANCE: This study will be conducted in compliance with the Good Laboratory Practice (GLP) regulations cited above. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. Tanhde wQiulalliintsypAesctsucrriatinccalepUhnaitse(sQoAfUt)hewilstauuddyitinthaeccporortdoacnolc,etwhiethrtahwedSattaanadnadrdthOeperreapotritn,g Procedures of Argus Research Laboratories, Inc. TachceurfaintaellyrerpeofrltecwtislltihneclruadwedaatsataotbetmaeinntedsidgunreidngbtyhtehpeeSrtfuodrymaDnicreecotforththeastttuhdeyraenpodrtthat all applicable GLP regulations were followed in the conduct of the study. Should significant deviations from GLP regulations occur, each will be described in detail, together with how the deviation might affect the quality or integrity of the study. 000664 418-008:PAGE F-3 Protocol 4P1a8g0e83 STUDYSCHEDULE: `See ATTACHMENT 1 to the protocol. TA ESTRTIANC D VEL HICE LE: Identification: Test Article Name: Physical Description: ~~ PFOS. Light-colored powder Lot/Batch Number: Specific Gravity: 217. 06. Purity: Expiration Date: 98.9% May, 2000. Information on the identity, composition, strength and purityofthe test article is on file with the Sponsor. Vehicle: 0De.i5o%niTzweedeWnat8e0r)i.n RSeuvpeplriseerdaOnsdmloostiisdenMteifmibcartaionneofPrTowceeesnse8d0DteoiobneidzeodcuWmaetnetre(dR.inO.the raw data. NtoeibteheprretsheenStpionntshoervneohrictlheetShattudwyouDlidreicnttoerrfiesraewwairthe tohfearneysuplottsenotfitahliscosntutdaym.inTahnetrsefliokreel,y no analyses other than those mentioned in this protocol will be conducted. Safety Precautions: Gfolromvuelsa,timoanskpr,epaaprpartoiporniaatnededyeospargoteecatdmiionniastnrdataiounn.ifoTrhme/lMaabtecroiaatlaSraefetotybeDawtoa mShdeuerting (MSDS) is attached to the protocol (ATTACHMENT 2). Storage: Bulk Test Article: VPerheipcalreeCdoVmephiocnleen:ts: Prepared Formulations: Room temperature. RRoooomm tteemmppeerraattuurree.. Frozen (-20C). AJullitaenstGaurltbiicnlseksi,hiMpamnenatgsertootfhFeoTremsutliantgioFnasc,ilattythsehopurledviboeusaldydcrietsedseadddtroetshse attention and of telephone number. 000665 418-008:PAGE F4 Protocol 418.008 Page 4 Shipments should include information concerning storage conditions and shipping csahritpomnesnts.hould be labeled appropriately. The recipient should be notified in advance of FORMULATION: EreqoufPreepnaractioyn: Formulations (suspensions) will be prepared daily at the Testing Facility. 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. TestingFacility ReserveSamples under the previously cited conditions The Sponsor will reserve a sample (1 g) of each lotofthe bulk test article used during the course of this study. The Testing Facility will reserve a sample (5 mL) of each lot of the vehicle components used during the course of this study. Samples will be stored ANALYSES: `thSeamcpoluersseadodfittihoenasltutdoy.those described below may be takenif deemed necessary during Bulk Test Article Sampling: No analyses of the bulk test article will be conducted during the course of this study. Information on the stability of the bulk test article is on file with the Sponsor. Analyses of PreparedFormulations: Stability: `Stability data for prepared formulations bracketing the range of concentrations and conditions of this study are on file with the Sponsor and will not be determined during the conduct of this study. Suspensions will be prepared daily at the Testing Facility. 000666 418-008:PAGE F-5 Protocol 418-008 Pages HomogeneityAnalvses: cHooumrosgeeonfeithtiysosfttudhye. teAstsayrrtiincglee iwnilplrbeepaurseeddstuoswpietnhsdiroanwsswaimlpbleesve(r5ifmieLd deuarcihn)gftrhoem the tEoapc,hmisdadmlpelaen(d5 bmoLt)iowimlolfbethdeivhiidgehdesitntcootnwceonatlriaqtuiootns,oonntehoe ff2irstmdLayanodf oprneepaoraf3tiomnL.. aOtntehealTieqsutoitn(g2 FmaLc)ilwtiylabseasbhaicpkpuedp fsoarmpalneal.ysiBsa;ctkhueposthaemrplaleisquwoitll(b3emsLt)orweildl ubnedreertatihneed previously cited conditions and discarded at the Testing Facilty upon the requestof the. Sponsor. `Concentration Analyses: oCfontchiesntsrtuadtyi.onoAfstyrhienpgreewpialrebde tuessetdatrotiwclietshudsrpaewnssiaomnpslewisll(5bemvLereiaficehd)dfurroimngetahcehcourse (c5onmceLnteraacthi)onwidllubreingditvhiedefdirsitnatondtwsoixatlhiqwueotesk, oofndeoosaf2gemLadmainndistornaetioofn.3 Each sample mL. One aliquot T(2esmtLi)ngwiFlalcbiletyshaisppaebdafcokruapnaslaymsipsl;e.theBaotchkeurpalsiaqmupotle(s3 wmiLll)bweilsltboererdetuanidneedr tathethe previously cited conditions and discarded at the Testing Facility upon the requestofthe Sponsor. `ShippingInstructions: `Samples to be analyzed will be shipped (frozen on dry ice) to: Kris J. Hansen, Ph.D. 3M Environmental Technology and Safety Services 9Bu3i5ldBiungsh2-A3v6e-n0u9e St. Paul, Minnesota 55133-3331 TTeelleefpahxo:ne: ((661122)) 777788--66011786 Both the recipient and the Study Monitor wil be notified in advance of sample shipment. DISPOSITION: Prepared article wil formulations be returned twoillthbeeSdtiusdcyarMdoenditaotrtahtetTheestpirnegviFoaucsilltyyc.iteAdllardedmraeisns.ing bulk test 600667 418-008:PAGE F-6 Protocol 418-008 Pages TEST SYSTEM: `SpecianedRsea/soSn ftorrSelaecitionn: TbehceauCsre::CD1)BtBhRisVsAtrFa/iPnloufsrat(hSapsrabgeueen-Ddaewmloenys)trraattweadstosbeleecsetnesditaisvethteo TreesptroSdyucsttievme aTenpdroddeuvcetliovpemaenndtadletvoexlionpsmaenndtahlatsoxbiecietny ewviadleulaytiuosnesd; 2t)hrhoiustgohroiuctalidnadtuastarnydfoerxperience exist at the Testing Facility; and 3) the test article is pharmacologically active in the species and strain. Number: IPnoiptiuallaptoipounlasteiloenctaecdclfiomrastteudd:y: 117955 vmiarlgienrmaatsle(3a5npder20d5osviarggiengfreomuapl)earnadts.175 female rats (35 per dosage group). Tpernesmuamteeddgfeestmaatlieonr;attshewilrlebmaeiansisniggfneemdatloeCraaetssawrilelabn-espeecrtmiiotntiendgtoondedlaiyver10litoefrs. 250 ay F1 21 generation postpartum pfuorpcso(n2t5inpueerdspeoxstpneartadlosobasgeervgartoiounp.) will be selected at weaning on BWode y iganhdAgte: Male rats will be ordered to they will be expected to be weigh from 300 at least 60 days g to 325 of age. g each Female at receipt, at which rats will be ordered time to wleeaisgth6f0rdoamy2s0o0fgagteo.22A5ctguaelabcohdaytwreeciegihptts, waitllwhbiecrhetciomredetdhetyhewildlabyeafetxeprercetceedipttoabned awtil be documented in the raw data. The weight ranges will be included in the final report. sex: mBaotlhe FaondanfdemFa1legernatesrawitlilobnemgailveenantdhefteemsatlaertriacltes.will be evaluated. Only Fo generation Source: Charles River Laboratories, Inc., Raleigh, North Carolina. The rats will be shipped in fitered cartons by air freight andor truck from Charles River Laboratories, Inc., to the Testing Faciity. 000568 418-008:PAGE F-7 Protocol 41P6a.g0e0?8 Identification: EGoeneration: RCoa.t,s Ianrc.e, pNeor.maMnSePntTly20i1d0e1n)t.ifieMdaulseinagndMofnemeallesrealtfs-paireercaisngsieganretdatgesm(pGoeryarByannudmabnedrsTaatg receipt before aadnmdingiisvterantuionniqouftehepefrirmsatndeonstaigdeenotfiftichaetitoenstnaurmtibceler.s when assigned to the study F1/F2 Generations: Pinutpesrwmisllonfotthebelititnerd.iviAdtuawlelayniidnegn,tifeiaecd hdurratinsgelleaccttaetdiofno;r aclol nptairnaumeedtoebrssewrivlaltbieonevwialllubaeted identified with a Monel selt-piercing ear tag. ANIMAL HUSBANDRY: All cage sizes and housing conditions are in compliance with the Guide for the Care and UseofLaboratory Animals. Housing Fo Generation Rats/F1 Generation Litters: eFxocgeepntedruartiinogn rtahtescwoihlalbbietaitnidoinviadnudallpyoshtopuasretduminpsetraiiondlse.ssDsutreienlgwicorhea-bbiottattoiomne,d ecaacghespair of rats will presumed be housed in gestation, Fo tgheenemraalteiornatf'semcaalgee.ratBsegaisnsniignngednotloantaetrutrhalandedliavyer2y0wiolfl be individually housed in common nesting box nesting boxes. Each during the postpartum dam and period. delivered litter will be housed in a F1 Generation Rats/F2 Generation Litters: Ahfotuesrewdeainnipnagi,rst(hoenFe1mgaelneerraattipoenr rfaetmsawlilelrbate)idnudriivnigduaclolhyabhiotuatsieodn,beafnodreincdoihvaibdiuatlaltyion, hFoougseenderaafttieorncorhatasb.itaBteigoinn.niTnhgensoalmateertytpheanofdacyag2i0ngofwilplrbeesuumseedd gaesstdaetsicorn,ibed for the dFe1ligveenreerdatliitotenr fwielmlablee hroatussweidllibneaicndoimvimdouanllnyehsotiunsgedboixn dnuersitnigngtbheoxpeoss.tpEaarcthumdpaermioadn.d 000669 418-008:PAGE F-8 Protocol 416-008 Pages Nesting Material: Bedding material (bed-o'cobs) will be supplied to female rats assigned to natural delivery. ABneadldyisnegswiflolrbpeoscshibalnegceodntaasmoifntaetnioans anreececsosnadruyctteodkeaennputahlelyanainmdaldsocdurmyeanntdedcleinanthe raw data RT ooe m Aim r, peraat ndu Humr idie ty: Tfrheeshanaiirmtahlatrohoasm ibseienndeppaesnsdeednttlhyrosuugphpl9i9ed.9w7it%hHatElPeAasftiteernsc(hAiarnogCelsepaenrhroouorm)of. 100% cRoonsotmanttelym.perRaotourmehwuimlidbietymawiinltaalisnoedbeatmo6n4itFor(e1d8cCo)nsttoan7t9lyFa(n2d6mCa)inatnadinmeodniatto3r0ed% to 70%. Light Amaninatuationmeadt.icaElalcyhcodnatrrkolpleerdi1od2-whiollurbleiggihtn:a1t2-1h8o0ur0 dark fluorescent hours EST. ight cycle will be Diet: Rats will be given Certified Rodent Diet #5002 (PMI Nutrition Intemational) available ad libitum from individual feeders. Water `aWnataeurtowimlaltbiec awvaatielraibnlgeaadccleisbsitsuymsftreomm. iAnldlivwiadtuealr bwioltltlbees fartotmacaheldoctaol tshoeurccaegeasndofpfarsosmed tphrroocuegshseadrweavteerrseaossamobsacitsermieosmtbatr;apnreocbeesfosreed uwsaet.erChisloerxipneectwieldl btoecaodndteaidntnoothmeore tbahcatner1i.a2l pcopnmtacmhilnoaritnieonatatnhde ttwiimceeoafnannuaallylsyisf.orWpaostseirblies acnhaelmyizceadl mcoonnttahmliynaftoiropno.ssible Contaminants: tNoeibteheprrtehseenStpionntshoercnerotriftiheed dSiteut,dythDeirdercitnokrinigs awwataerreoorftahneynpeostteinntgiamlacteornitaalmiatnalnetveslslitkhealyt would interfere with foutinely performed bthyetrheesufletesdosfutphpilsiestruodyr.thTohseeremfeonrtei,onneodaninalthyissesproottohceorltwhialnl btehose conducted. 600870 418-008:PAGE F-9 Protocol 41P8a-g0e089 FGoeneration: gUepnoenraatrreidvarl,anradtosmwiulnlibtse.aAsfstiegrnaecdcitiomiantdiiovni,dumaallheouasnidngfeomnalteherabtassiwisllobfecosmepluetceterd- for Study on the acclimation. basis of The rats pwhiyllsibcealaasspipgenaerdantocedoasnadgbeogdryowuepisgbhatsserdecoonrdceodmpduutreirng-generated (weight-ordered) randomization procedures. Wcoihtahbiintaetaicohn,doonseagmealgeroruapt,pceornfseemcaulteivreato.rdTehrewiclolhbabeiutasteidontopearsisoidgnwilrlatcsontosist ofa mvaagxiniamlucmonotfen1t4sdaaynsd.orFeamcaolpeurlaattosrwyitphlusgpoebrsmeartvoezdoainosbistuerwivleldbein a smearof considered the to be at dmaayte0d owfitphrienstuhmeefdirsgtes7tdataiyosn oafncdohaasbsiitganteiodntwoililndbieviadsusalighnoeudsianlgt.ernFaetmeamlaelerartastsnotthat have mated (same dosage group) and will remain in cohabitation for a maximum of seven additional days. Tahsseifgrnsetdtteon Cfaeemsaalreearnat-ssepcetridoonsinaggeongrdoauyp 1w0itohfaprceosnufmieneddgedsattaetioofn.matTihnegrweilmlabineing female rats will be permitted to naturally deliver litters. Astaambpllee ocfolrlaecntdioonmautnistcshweidlullbeedussacerdiftioceasasfitegrncfoimveplreattsiopnerofgtrhoeupcothoabaiptahtairomnacpoekriiondetic p(moaslteparrattusmsi(rfiengmalliteterrastwsiathlldoawemdstaolnlaotwuerdaltloy naturally deliver deliver litters). a litte) or on day 21 E1IE2 Generation Pups: Day 1 which of all lpaucptastiionna(lpiottsetrpaarretuimn)diivsiddueaflilynewdeiasghtehde day of birth and is (pup body weights aso will the first day be recorded on after all pups in a liter are delivered and groomed by the dam). Oannddlaityte4rspwoilsltpbaertruemd,ucaedtatboleeiogfhrtapnudposmeuanciht.s wWilhlebneevuseerdpotosssieblleec,ttphuepssatmoebenucmulbleedr.of male and female pups per liter will be continued on study. AwitllwbeeanuisnegdotfotsheeleFc1t g2e5nemraalteioanndpu2p5sfoenmadlaeyp2u1ppsopsetrpagrrtouump,, aretsaubllteinogfirnaantdootaml uonfit2s50 Fp1upgeannedraotnieonfermatasle(1p2u5pppeerrsleixt)erc, hwohseenn for continued possible, will evaluation. be selected. At least one male 000671 418-008:PAGE F-10 Protocol 4Pa1g8e-0180 ADMINISTRATION: RoR utee andasfoo rChon ice: Tdiheetaorryalro(ugtae,vatghee)erxoaucttedwoassagseelceactnebdefoarccuusreatbeelcyauasdmei:ni1s)teinrecdo;mapnardi2s)onitiwsitohntehoefthe possible routes of human exposure. MeatndFhreqo uend cy: `Daopsparogxeismawtiellybethaedsjuasmteedftiomretheeacmhodsaty.recently recorded body weight and given at EGoeneraMtalieRoatns: M(amlaexiramtusmwil1l4bdeaygsi)veanntdhceotnetsitnuaritnigcltehornocueghdatihleydbaeygibnenfionrge 2s8acrdiafyicse.beMfoarleecroahtasbwiitlal tbieon sacrificed after completion of the cohabitation period. Eo GenerationFemaleRats: cFoehmaabliteatriatosn w(ilmlabxeimgiuvmenofth1e4tdeasytsa)rtaicnlde oconncteinduaiinlgy btehgrionungihndga2y89odafysprbeesfuormeed gaessstiagtnieodn t(orantastausrasligdneelidvteoryCtaheastardeoann-ostedcetliiovneirnag)l,ttdera)yo2r4daoyf p2r0epsousmtepdargteusmta(triaotns t(hraatts deliver a liter). 1 Generation Fe1xpgoesneedrattoiotnheptuepstsawritlilcnloetdubreindgirmecattlyergiavlengetshteattieosnt (airntiucltee,robuetxpmoasyurbee)poofsvsiiablmyatemal milk during the lactation period. Dosages will be selected by the Sponsor on the basis of previous studies conducted with the test article. 000672 DC osaogenLevcelse, ntratanidVooln umess: 418-008:PAGE F-11 Protocol 4Pa1g8e00181 Ts[on[om [5lecsmssonmemvenl TColTes[[woeeT To om w T+Tousoooomumenen] |Jeoeomeomuoman] Thetestricevilbmconsire 100% uefor he pos of otagecaciaons TM ESTSE ,ANAA LYSS ESAU ND REME-FoN GENT ERATS ION: Viability - Male and F : All Periods: Atleast twice daily. Clinical Observations and/or General Appearance - Male and Female Rats: Acclimation Period: Atleast once. Dosage Period: Tawppircoexdiamialty.elyProionrethooduorspaogsetdaodmsiangies.tration and once Maternal Behavior: aDbanyosrm1,a4l,b7e,ha1v4iaorndwil2l1bpeosrtepcaorrtduemd. daAilny.y observed CalpipnriocparlioabtseerbvyatthieonSstumdayyDbireecrteocroardnedd/omroSrteudfyreMqouneinttolry.than cited above, if deemed Body Weights - Male Rats: Acciimation Period: Atleast once. Dosage Period: Weekly. Sacrifice Terminal weight. 000673 ` 418-008:PAGE F-12 Protocol 4P1a8g.e00128 BWodyeig-Fhemat leRs ats: Acclimation Period: At least once. Dosage Period: Weekly to cohabitation. Daily during presumed gestation and on Days 1, 4, 7 and 14 postpartum (rats assigned to natural delivery). Sacrifice: Terminal weight. EeCedonsumVapluets-iMaloeRants (recorded and tabulated) Dosage Period: Weekly. FCeedonsumValpuest-FeimaloeRants (recorded and tabulated): Dosage Period: Weekly to cohabitation. Daily during presumed gestation. Days 1, 4, 7 and 14 postpartum (rats assigned to natural delivery). Feed consumption not tabulated after day 14 postpartum, when it is expected that pups. will begin to consume maternal feed. Eeed Consumption Values - Male and Female Rats Feed neces consum sary to ption values replenish the mfaeeyd.beDrureicnogrdceodhamboitraetiforne,quwehnetlny than cited aboveifit two rats occupy the is same cage with one feed jar, replenishment of the feed jars will be documented. Individual values will not be recorded or tabulated. Estrous Cycling and Mating: A table of random units will be used to select 15 female rats per group for evaluation of ecoshtarboiutsactyicolninpgerbiyodexamination of vaginal cytology for 14 days before the start of the During cohabitation, all female rats will be evaluated daily until spermatozoa are observed in a smear of the vaginal contents and/or a copulatory plug is observed in situ. DuraoftGesitaotionn: The duration of gestation is calculated from day 0 of presumed gestation to the day the first pup is observed. 000674 418-008:PAGE F-13 Protocol 4P1a8g-e00183 Fertility Parameters: Fentiity Index (percentage of matings that result in pregnancies). Gestation Index (percentage of pregnancies that result in bith of live liters). Number of offspring per liter (ive and dead pups). Number of implantation sites. General condition of dam and liter during the postpartum period. Viability Indices (percentage of pups bom that survive 4 and 7 days). Lactation Index (percentage of pups bon that survive 21 days) Caesarean-Sectioning Observations: Rats will be Caesarean-sectioned on day 10 of presumed gestation. Placentae that appear abnormal (size,coloror shape) will be noted in the raw data. The rats will be examined for number and distribution of Corpora Lutea. Implantation Stes. Viable and Nonviable Embryos. (A viable embryo is oval or crescent shaped, pink, fin and enclosed in an p`aimnnkiottoitcasnaocrfdileleedpwirtehd ctolebalrafclku,id.sofAt annodnveinabclleoseemdbirnyaonisamanmioortpihcosuasc,fsimlaeldlw,itphale clear, cloudy, or opaque fluid.) Natural Delivery: Female rats will be evaluated for: Clinical Observations During Parturition. Duration of Gestation (day 0 of presumed gestation to the time the first pup is. observed). Length of Parturition (time of first pup divided by N-1 pups idneleiavcerhy loiftelra).st pup minus the timeofdeliveryofthe Litter Size (defined as all pups delivered). 600675 418-008:PAGE F-14 Protocol P1a8g.e0018a Pup Viability at Birth. MS ETHA ODOC F RI-FFoGIENEC RATIE ON: eRxaatmsiwnialltiboensacrificed by carbon dioxide asphyxiation. Embryos will be discarded after NECR- FoOGENPERSATIYON Sex from which all issues examined at necropsy will be retained, in order to provide Gross lesions will be retained in neutral buffered 10% formalin for possible future `evaluation (a table of random units will be used to select one control group rat of each control tissues for any possible histopathological evaluationsofgross lesions). Unless specifically cited below, all other tissues will be discarded. Male and Female Rats Assigned to Pharmacokinetic Sample Collection: the Sponsor for analysis. At scheduled sacrifice after completionof the cohabitation period (male rats siring litters with dams allowed to naturally deliver a litter) and on day 21 postpartum (female rats allowed to naturally delivera litter), five rats per group will be assigned to a pharmacokinetic sample collection. In addition to the appropriate evaluations described below, blood samples (approximately 4 mL per rat) will be collected from the inferior vena cava into serum separator tubes and centrifuged. The resulting serum (approximately 2 mL) will be immediately frozen on dry ice and maintained frozen (-70C) until shipment to the Sponsor for analysis. The liver will be excised, weighed, and a sample section (lateral lobe) will be frozen and retained at -70C until shipment to Aft be er sh completion of ipped (frozen sa on mple col dry ice) lection, to Kris serum and J. Hansen, liver section Ph... at the (lateral lobe) samples will previously cited address for analysis. Both the recipient and the Study Monitor will be notified in advance of sample shipment. Sch ifice of Male Rats: After completion of the onercgraonpsswyilofbteheextchiosreacdi cohabitation ca,ndabwdeoimgihneadl peri aanndd od, male rats will be sacrificed and a gross rpeetlaviicnevdisfcorerpaoswislilblbee hpiesrtfoolrogmiecd.evaTlhuaetfioolnl:owing testes, epididymides, prostate and seminal vesicles (weighed with and without fluid). nTehuetrtaelsbtuefsfweirlledbe10fi%xefdorinm Bouin's alin for solution possible for 48 to 96 hours histopathological and then re evaluation. tained in The remaining organs will be retained in neutral buffered 10% formalin. 600676 418-008:PAGE F-15 Protocol 4P1a8g-e00185 chedul rifice - Feral ignedto C: an-Sectioning: On day 10 of presumed gestation, female rats wil be sacrificed, Caesarean-sectioned, and a gross necropsyofthe thoracic, abdominal and pelvic viscera will be performed. cUotenrfiiorfm tahpepaarbenstelnycenoonfpirmepglnaanntattriaotnsswiitllesb"e. sUttaeirnieodfwnitohnp1r0e%gnaanmtmorantisuamndsuaillfiodveatroies will be retained in neutral buffered 10% formalin for possible future evaluation. h fice - Assi I Delivery: Rats that do not deliver a litter will be sacrificed on day 25of presumed gestation and examined for gross lesions. Uteri will be stained with 10% ammonium sulfide to confirm the absenceof implantation sites. `After completion of the 21-day postpartum period, female rats will be sacrificed, and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. The number and distribution of implantation sites will be recorded. DSamuswirthvNoivinPupgs: Dams with no surviving pups willbe sacrificed after the last pup is found dead, missing or presumed cannibalized. A gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. Postpartum data for these dams wil be excluded from summary tables. Rats Found Dead or Moribund: Rats that die or are sacrificed because of moribund condition or abortion will be meaxdaem.ineTdheforratthsewicllaubseeeoxfadmeiantehdfoorrmgorroissbunldescioonndsi.tiToenstoens,theepdidaiydythmeidoebss,erpvraosttiaotneisand seminal vesicles of male rats will be recorded (seminal vesicles weighed excised with and and individual without fluid). organ weights will The testes will be be fixed in BTohueinr'esmasionluitnigonorfogran4s8 wtioll9b6ehroeutrasinaendditnhneenutrreatlaibnuefdfeirnedne1ut0r%alfbourfmfaleirne.d 1Pr0e%gfnoarnmaclyin. dsetlaitvuseraenddpuutpesriwnilelcboenteexnatmsionfefdemtoalteheraetxstweinlt pboessriebcloer.dedO.varAibeosrtweildlbfeetruesteasinaendd/ionr neutral buffered 10% ammonium 1su0l%fidfeortmoalcino.nfiUrtmertiheofaabpspeanrceentolfy inmopnlpanrteagtniaonntsirtaetss.will be stained with 000677 418-008:PAGE F-16 Protocol 4P1a8g-e00168 TM ESTSE .ANA ALYSS ESAU ND REME -F1N GENET RATS ION: Viability: Preweaning Period: dLiatyte.rs Twhilelbpeuposbsienrevaecdhfotr rdeawidlpbuepscoautnlteeadstotnwciece Postweaning Period: Twice daily. CO linib cal servanda lor t GeneiralAoppen aransce: Preweaning Period: Once daily. Postweaning Period: Once weekly. Maternal Behavior: `Daabynsom1,al4,b7e,ha1v4ioarndwil2l1bpeosrtepcaorrtduemd.daAilnyy. observed CalpipnriocpalriaabtseebryvatthieonSstumdayyDibreecrteocroarnddeldormotrhee fSrteuqduyenMtolnyittohra.n cited above,if deemed Body Weights: Preweaning Period: Postweaning Period: Days 1 (birth), 4,7, 14 and 21 postpartum. Weekly Presumed Gestation Period: Lactation Period: Sacrifice: Days 0. 7. 10, 14, 17 and 20 (female rats only) Days 1,4, 7, 10 and 14 (female rats only). Terminal weight Eeed Consumption Values (recorded and tabulated) Preweaning Period Not recorded. Postweaning Period: Presumed Gestation Period: Lactation Period: Weekly except during cohabitation Days 0,7, 10. 14, 17 and 20 (female rats only). Days 1, 4,7, 10 and 14 (female rats only) 000678 ` 418-008:PAGE F-17 Protocol 4P1a8g-e00187 frFeeeepelddejnacirso,hnvstauhlmeupeftseiewdoi.lnl vbDaeulrudieonsgcmucmaoehyanbbtieetdartewicohnoe,rndwefhdeeemndortjawerosfrraeartqesuefoniclteclduy.ipfyTthtihesesneseacimnetseesrcavaralygsetwoiwliltnhootnbee tabulated. Preweaning Developmental Observations: cTohnetinnuumebseurnotifl tphuepdsamyetehteincgrittehreiocnriitseraitotnaiinserdecboyradleldpuopnseiancthhedaliyteor.f testing. Testing Surface Righting Reflex (abilty to right in 5 seconds): From day 1 postpartum. Pinna Unfolding: From day 2 postpartum. Eye Opening: From day 12 postpartum. Acoustic Startle Response: From day 13 postpartum. Air Righting Reflex: From day 14 postpartum. Pupil constriction is evaluated once, on day 21 postpartum. Postweaning Developmental Observations: `Sexual Maturation: opFonesmdtapalayret3ur9mat.psoswMitlpalalbrteurematv.salwuialtbedefeovratlhueataegdefoorf tvhaegiangael poaftpernecpyu,libaelgsienpnairnagtioonn,dbaeyg2in8ning Passive AvoidanceTesting: rwBehetgeeirnntenioipnnosgasniabtdl2el4,onw+gil-1ltebdreamyervpeaotlesunttapitaoenrd.tuimn,aopnaessmiavleearvaotiadnadncoenteesftemfoarlelearartnifnrgo,msehaorcth-tfietrtemr, oPTtlhheeexripgaclsoasmsipvaelridats.vmoeinOdtnaensccfeiotmateppdpaawrrittamhteuanstgcirsoindfsfiiltsoetodsr wotioftwahhtaiwcbohr-iacghobtmrpiieagfhrt(t1maesnendct)Pclpheuaxlimsgbeleoarfsmwiiftllohdoreh.liencTgthereidc idgsouororrp.eenntOe(nd1 eamanAcd)htcthaeenstlbitgerhitald,iesltithvueermreeadtd.ioTnp.hleaTcthewdeoircnattoomitsphaaelrlt"obmrweiengdhttst"oaceroxempplsaoerrpteamrteahntete,adptbphyaeraasltsiuldisidniugnngtdiooftr rtgneutmmeoervdseotdfhfefar"ondmdarttkhh"eecaborpmippeaafrrpatutmiuessneta.onfdcTuphrleraecnseltididsiinndgteoldiaovoehrroelidsdittnhogetnchaeigmgemreifddoirfal3oto0er.lsyeTcchloeonsdersadt,biteshfteohrleeingthhteis start of the next trial. Trials are repeated until the rat remains in the "bright N 000679 418-008:PAGE F-18 Protocol 4P1a8g-e00188 mc15oamxtpriiamarlustmmhea6nv0te-sfboeerce6on0ndcsoeimncptolenrevdtaesldo.isnrTtehwceoorcldaoetndesnefccoyrutteoiaevcnehttertrriiatallhs.ethdearckricteormiponarfotrmelenatronrintgh)eor until tEhaecchrirtaertisonteissttehdetswaicmee. fTorhebottehstdsaeysssiofontsestairneg.separated by a one-week interval, and Dosage groups are compared for the following dependent measures: tTohceonmupmabreergorfoutrpisalsfotrootvheeraclrlitleeriaomniningtpheerffiorsrtmsaenscsei.on--this measure will be used `Tccohomempplaaarrtteemngecrynot(uionpnssetfcoiraolnacd1tsii)vnitttyoheelenfvtiereslrtsttaehnsetd"sdeeaxsrpskli"oorcna--ottmohrpiyasrtmteenmdaeesnntucrifeeroswmilinlthabeen"oubvrseielgdhtt"o environment. Tccoohmmepplaaarrtteemngecrynot(uiponsnsetfrcoiroalnsh2dosri)nt-tttoheeernfmtirersretttteehnsetti"osnde.asrski"onc-o--mthpiasrmtemaesnturferowmillthbee"ubrsiegdhtt"o Tbeheusneudmbtoercoomfptrairales gtorotuhpescrfiotrerlioonng-intetrhme rseetecnotniodn.test session~this measure will clToohnmegp-latarettremmnecrnyett(eionntnsioetnrci.oaln1dsi)n ttoheensteecrotnhde s"edsarski"onc--otmhpisarvtamleunetisfarnoomtthheer"ibnrdiigchatt"ion of WatermazeTesting: aBebeaigclithyn,lniiltneeagrrwanitilnagbpeparneodvxaimlmeuamatoterelydy.i7n0adwaaytserp-ofsitlepdarMt-umm,azoneefomraolveerrtatcoaonrddionnateiofne,maslweimramtifnrgom ifEsoarfcitlhelemrdpawteiritashttuwerasettee(drratinnogaaewdoaeftpe2tr1htiogCfhta+p11p6r-Cog)xa.iumgaetesltyainnilneessinsctheeesl,moadnidfitehde Mw-atmearzeis.moTnhietomraezde MOs-tnmemaezafeca,hrthitneeossrttdtfreriarolt,motthhbeeertarwteowmialolrvmbesed)pfalrnaodcmertdehqienutwioartetedhret.ostsOawrntiimtnhgteopfooisrnsitteitoroifnalt(, hbteahsetewroaotfgitosharelesMq-uoimfratezhdee.to cfeanhiltoetsroenbmoatokhneatraimalcso1rorifesctdtheegsoimagalnzacetheobdiecfteohreweiitnbhceioinrnr6ge0crtsegemocoaovlnedddusrfiirnnogamnttyhheegriwevaemtnaeirtn.riianlTgahrtereiiagnlisut.iiadlReaadtrsfmotthhaet secreorrporarerlcaettsgseoetaraliacalhsntdtroiaatl.reermEtiahnecanhterretathmieosvtreesedtqufsirersoesmdiottnho.erweTaathceehr.amacAxrii1tem5r-uisomneocnfoufnmidbveienrtceorontfrsitearcliauiltnstiievnrevaanlywitlelst 600650 418-008:PAGE F-19 Protocol P41a8g-e00189 mseasxsiiomnumis 6105.-sLeactoenndciynt(emrevaalsuisrreedcionrdseedcofnodrse)actohctrhioalo,saestihsethceornruecmtbgeoralofoerrtrhoers (incorrect tums in the maze) during each trial. Each ratis the correct gtoeastleadntdwitceh.e cTrihteertieosntarseestshieonssaamree fsoerpbaortahtetedstbyseassoinoen-s.week interval, and Dosage groups are compared for the following dependent measures: `uTsheedntuomcboemrpoafrteiaglrsotuopscrfiotreroivoenroalnltlheearfnirisntgdpaeyroffortmesatnicneg.--this measure will be `fiTrhste daavyeroafgteesntuinmgb--ethrisofmeerarsorusre(inwiclolrraelcstotbuemsusiendthtoe cmaozmep)arfeorgeraocuhpstrfioalroonvertahlel leaming performance. tTehsetilnag--ttehnicsy (mienasseucroendwsil)l tboeruesaechd ttoheccoomrpraecrteggoraoluopns tfroirals2hoofrtt-hteerfmirrsettdenatyioonf. bTeheusneudmbteorcoofmptrairales gtorocuriptserfioornloonngt-hteersmecroetnedntdioany.oftesting--this measure will m`Tehaesauvreerawiglel anlusmobbeeruosfeedrrtoorscofomrpeaarcehgtrroiaulposnftorhelosnegc-otnerdmdraeyteonftitoensting~this The latency testing--this i(sinasneoctohnedrsi)ndtiocarteoarcohftlhoengc-otrerercmtrgeotaelntoionntrial 1 of day 2 of Reproductive Capacity: Abte aapspsriogxniemdatteolcyoh9a0bidtaaytisono,f aognee, mtahleeFr1atgepneerrfateimoanlerartast,wbitahsiendeaocnhcodmopsuatgeer-ggreonueprwaitlled croahnadboitmatuinointspoerriroadnwdilolmcounnististtabolefas,mwaitxhitmhuemexocfl1u4sidoanyso.f siFbelimnaglmeatriantgssw.ithThe osbpseerrmvaetdozionasiotbusweilrlvbeed cionnasisdmeeraerd otfo tbheeavtagdianyal0coofnptrenetssuamnedd/ogresatactoipounlaatnodryaspsliuggned mctoaothieanbddi.itvaitdFiueaomlnahwloieulsrbiaentgsa.wsisFliegbmneaedlaelallrotawetersdntathtoaetnmadatoulrenaolrtlaytmsdaeftlreiovwmeirttahhienndstamhameiefnitrdsatois7nadlgiaetytegsrsrootfuhprotuhgaht ahave 21-day postpartum period. MatingPerformance: As cited above for Fo generation rats. 600681 418-008:PAGE F-20 Protocol P41a8g.e02008 DuraoftGesitaotionn: As cited above for Fo generation rats. Eertility Parameters: As cited above for Fo generation rats. EG2eneraLitttier Doatna: aVisabciiltiteyd,acblionvicealfoorbFs1ergveanteiroantsiaonndlibteordsy. `weights for F2 generation pups will be recorded D ACH =F: \TION As previously cited for Fo generation rats. NECROPSY - F1 GENERATION RATS: RATS/F2 GEN TION PUPS: eGvraolsusatlieosnio(nastwaibllleboefrreatnaidnoemd uinnintesutwirlallbbeufufseerdedto1s0e%lecftoromnaleincofnotrroplosgsriobulpe fruattuorfeeach `sex from which all control tissues for tissues examined at necropsy any possible histopathological will be retained, in order to provide evaluations of gross lesions). Unless specifically cited below, all other tissues will be discarded. Scheduled Sacrifice - F1 Generation Male Rats: Rats will be sacrificed after completion of the 14-day cohabitation period. A gross necropsy of the thoracic, abdominal and pelvic visceral will be epididymides of male rats will be excised and individual organ performed. weights will Testes and be recorded. formalin, The epididymides will be retained in fixed in Bouin's solution for 48 to 96 neutral buffered 10% formalin. The testes hours and then retained in neutral buffered will be 10% Scheduled Sacrifice - F1 Generation Female Rats: Female intuemrbweilrl rats will be sacrificed after completion of the 21-day postpartum abnedsadcirsitrfiibcuetdioonnodfaiymp2l5anotfatpiroenssuimteesdwiglelsbtaetiroencoarndedd.uterRiatwislltbhaet period. sdtoainnoetd The dweilthivear n1e0c%roapmsymoofnithuemtshuolrfaicdiec,toabcdonofmiirnmalthaenadbpseelnvicce voifsciemrpalawnitllatbieonpesriftoersm.ed.A gross Female rats `without a confirmed estimated day 25 of mating date that do not presumed gestation. deliver a litter will be sacrificed on an 000682 418-008:PAGE F-21 Protocol P41a8g-e02018 Fo ribund: m`Reaaxdtaesm.tihnaTetdhdefioerroattrhsaewricelal usbasecereoixffiacdmeeidantbehedcoafrourmsogerroiofsbsumnoldersiciboounnsdn.idticTooennsdtoientsi,tohneepoidrdaaiybdoytrmhtieidooenbss,weilrplvroabstetiaotneisand Brsoeeucmioinrn'daseldsvoe(lssuietcimloiennsafloorfv4mes8ailcteloer9sa6twsehwoiiugllrhsbedeanwedixtcthihsaeennddraewntidatihinoneuddtivifinludinudea).ultorTarlhgeabnutfewfseetiregeshdtwis1ll0wi%bllefboferi.xmeadliinn dsTethlaeitvurseerameandidpnuiuntpegsriownirlegl acbnoenstewexinlaltmsbieonferfedettmaoainlteehderiaentxsnteewunittlrapbloesbsruiebflcfeoe.rrdeedOdv.1a0rA%ibeofsrortwimeladllibfne.efruePsteraseignanendadlniocnry n1e0u%traalmbmuofnfeiruemd 1su0l%fidfeortmoalcion.nfUitertihoefaabpspeanrecnetolfyinmopnlapnrteagtniaonntsirattess.will be stained with E1 Generation Dar ivi oDapmrseswiutmhendocasnunrivbiavliinzgedp.upsAwgilrlosbse nsaeccrriofpicseydoaffttheetthheorlaacsitc,puapbdiosmfionuanld adneaddp,elmviiscsing sviusmcmeraarwyiltlabbleesp,erformed. Postpartum data for these dams will be excluded from EAIF2 Generation P n stoartum: ssPitunapktuswsitlalhtabtbeidriitedh.ebnteTiffhoiereedleuaxnsagsmstiwinilallltbiobmoe:norpfeumptoshveweildtihtaelrnudfnogrismpmtuhepartvsifealdboialtintwyiwlawlitblereb.eidPeeunvtpailsfuiewaditteahdslfluoinrvgevsibtotarlhna,t esavonaldlututiaootnihoafnovsre,pdoitiswesidilbslbheeofrnutotyutreaedfteeivnralbtiuhrateht.nioencP.ruopSpshsoywuidtlahdtag.proosstsmloerstieomnsawuitlollbyseisprperseecrlvuedde itnheBosuein's EAIF2 Generation Pups Found Dead or Moribund on Days 2 to 21 Postpartum; flPoeusupinsodnfsoonaunndddayfdsoera2tdhteoor4caspauocsrseitofpifactrehdteudmumweoirtliolbbmueonrpdirecbosunendrdivtceiodonndiniotrBiodouenianwti'hlsl. sboPeluueptxisaonwmiiftnohregdprofosossrsibgllreoesssif.outnusre enevevauatllruuaaalttiiboounnf;sfiegtrrweoidslls1bl0ee%sniofoontrsemdaolfiinpn.uthpesShnfoeouculrndodpposonystddamatoyar.st5emtoau2t1olpyossitspparretculmudwiellthbeesepreserved in 600683 ' 418-008:PAGE F-22 Protocol 4P1a8g.e02028 foF1r garnodssF2legseinonesr;atpiuopnspwuiptsh cgurlolsesd loensidonas4ywilpobsetpparretsuemrvweildl ibneBsoaucirni'fsicseodluatnidonexamined hfNryeodcnrtroaoclp-epspayhraiwleitylal.linscultuudree aansdinegxleamcirnoastsi-osenocftitohnoefcrtohsesh-eseacdtiaotntehdeblreavienl foofrthaep.parent All F1 gross lgeesnieornas;tigornospsuplsesciuolnlsedwiolnbdeapyr2e1seprovsetdpianrnteuumtrwailll Necropsy will include a single cross-section of the head bbueffsearcerdifi1c0e%d afnordmaelxian.mined at the level of the. for frontal-parietal hydrocephaly. suture and examinationof the cross-sectioned brain for apparent ScheduledSacrifice-F2GeneratPiuposn: ONencrdoapys2y1wiplolsitnpcalrutduema, spiunpgslewiclrlobsse-ssaeccrtiifoinceodfatnhed heexaadmiantetdhefolrevgerloosfstlhees.ions. hfryodnrtoacle-ppahraileyt.al suture and examination of the cross-sectioned brain for apparent 600684 418-008:PAGE F-23 Protocol 4P1a8g.e0r0s8 PROPOSEDSTATISTICALMETHODS: I. Parametric TupeofTest' 1 . Nonparametrc* Aapvperropargieastea.ndAdpdeirtcieonnatlagpersocweildlubreescaalncdu/loarteadn.aLliytsteersvmalauyesbewiplelrbfeorumseedd,iwfhaeprpreopriate. A. Bartlett's Test" A. Kruskal-Wallis Test (s75% ties) Significant at p<0.05 Not Significant Significant at p<0.05 Not Significant Lo nlVariance JTest Significantat ps0.05 | Dunnett Test Not Significant ~~ B. Fisher's Exact Test (>75% ties) Il. TPesrtfooportiDoanta Variance Test for Homogeneity of the Binomial Distribution 2. b. Statistically significant probabilities are reported as either Used only to analyze data with homogeneityof variance. Ps0.05 or ps0.01, d.. PTreosptorfotriohnomdoagteanaerietnyootfivnacrliuadnecde,in this category. 0600685 418-008:PAGE F-24 Protocol 4P1a8g-e02048 DATAACQUISITION,VERIFICATIONAND STORAGE: oDDriairtgeiacntawoillrlraebncedo/rhodarsnadwpi-lplrabonpder/isoatrtocreoemmdpaiunntaetgrh-eermaeerccnohtridvpeeeds.rsoofRntneheceloTrwediststhiiwnnigl21Fbaecdiarlteyyvs.iaefAwtleeldrogrbeiyngeitnrhaaeltidSoatntu.adywAillll yrbeeeqaubreosatuf.tnedrPmaraneidsleiinrngvdoeefdxettdhi.sesdAureascfotwipflyilnabolferaselltpoorrrate,wdafdatatettrahwewhiTlilecsbhteitnsigumpeFpatlchiileeidtSytpoaotnthsneoorScphwoianlrsgboeerfcouorpnootnnaected to determine the disposition of these materials REC TOO BEMR AINTD AINS ED: TPersottoAcrotlicalen,dVAemhiecnldemeanndt/so.r Reagent Receipt, Preparation and Use. ARnainmdaolmiAzcaqutiisointioSnc.hedules. Mating History. GTerneeartamlenCto(mimf eprnetssc.ribed by Staff Veterinarian). BCllionoidcalSaOmbspelrevaCtoliloencstiaonnd,/PorroGceensesrianlg AapnpdeaShriapnmceen.t. FBeoeddy WCeoingshutmsp.tion Values. NCaateusraarleaDnel-iSveecrtyioObnsienrgvaOtbiseornvsa.tions. RLietftlerexObasnedrvPahtyisoincsa.l Development Gross Necropsy Observations. and Behavioral Observations - F1 Generation Pups. POhrogtaongWreaiphgsht(sif (rief qrueiqrueidr).ed). FSeteudd,y MWaaitnetreannadncBeed(drionogm Aannadlyesnevsi.ronmental records). Packing and/or Shipment Lists. KEY PERSONNE! DEixreeccuttoirvoefDRiersecetaorrchof: RAelsaenarMc.h:HoMbielrdmraedn,S.PhC.hDri.s,tiDaAn,BTPh.D., ATS DAisrseocctioartoefDLiarbeocrtaortoorfyROepseeraatricohnsa:ndJSothundyF. DBiraemcettotr:, BR.aS.ymond G. York, Ph.D., DABT MMaannaaggeerrooffASntiumdaylCoOopredriantaitoinosn:anVdalMeermiebeA.r,ShIanrstpietru,tiMo.naSl. Committee: Dena C. Lebo, V.M.D. Animal Care and Use CMoansnualtganeotr,f VReetgeurliantaorryyPCaotmhpolliogayn:ce:W.KRaathyleBernowAn.,MDo.rVa.nM,.,M.PSh..D., ACVP 00686 418-008:PAGE F-25 Protocol 418-008 Page 25 EINALREPORT: bAecfoimnpalriezhedenfsolilvoewidnrgafctofnisnualltraetpioornt following: will with bteheprSeppoansroerd.onThceomrpelpeotritowniolfl tihncelusdteudtyheand will E`xSpuemrmiamreyntaanldDCeosnicglnusainodn.Method. DAEapvtapale,unadPtiricooetnsoc:ooflTFeiagsnutdreRAses,ssuSoltcusim.amtaedryAmaennddImnedinvtisduaalndTaDbelveisatSiuomnsm,arSitzuidnygDitrheectAobro'sve `GLP QAU CSotmatpelmieanntc.e Statement, Reportsof Supporting Data (if appropriate) and INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: tIThnhasettiitpunrtvoiooclnevadeluAsrtneuisdmydaelasncCiramirabelesdanwiidnlltUhbsiesecpCoroonmtdmoucicottlteehdea.ivneaAblmeaeprnnonrceeervdiuteorweaesvdodibedsycotrrhiembieTndeismitinzitenh.gisFapcriolttoyc'osl discomfort, distress or pain to the animals. dnTuehpceleisScsapitotinyvsefoorsr'tcusodsnyidgmunacatytiunbrgeetbohebistlaosiwtnudeddoycfaurnmodemnttthhsee ftSahpceotnftsahocatrt.thhiaNstoiisnafnlootrtemmaaatntiiouvnnecn(oeincncevesirtsrnaoir)nigy the procedures were available for meeting the stated purposes of the study. 600687 418-008:PAGE F-26 Protocol 4P1a8g-e02058 REFERENCES: 1. ICTnherfstiossr.tmiaatEni,novnMi.SrSeo.rnvmaiencnedt,aVlUo.yPSt.reoktD,eecpPt.aiEro.ntm(Ae1g8ne8tn2c)oy.f,CIoWnamVsmihevironcgRete,opnrS,opdrDui.ncCgtf.iiveNeladta,inoVdnAaMlu2Tt2eae1gne6nn1iiccai!ty 2. nSCachrliitesrntecixeaosnn,,eMN.(oSP.rvoe(c1em9eb8d4ei)7n.r,gsR1e9o8fp3rN)oa,dluJt.crteCilxivonen.etPoSsxyiyccmihptiyoatsa.inud4m5t(,e9r)Na.t7eo-wl1o0Y.goyrekvaAlcuaat.dieomnsy ooff 3. ILCnoacnn.t,gr.WoilPlLDm.iant(ga1t9oi8nn8,)t.hMeAEChma0br1ri8ye8os7-aR0in6vd3e0Fre.Ctra(l:DDaCetDvaeblBaoRspemRaeptn.rtoavClihdaTerodlxibecsyitARyir(vgTeuersrLaRateobslooergaayrr)cihes, Laboratories, Inc.) 4. LaIbnsotritauttoeroyf LAanbiomraast.oryNaAtniiomnaall RAecsaoduermcyesPr(e1s9s9,6)W.asGhuiindgetofno,r tDh.eC.Care and Use of 5. SImaplleawnstkait,iEo.nss(t1e96l4l)e.n 247.367. aFmarUbteemreusthdoedreRaztutem. mAarkcrh.oskPoatphiosl.chEexnp.NaPchhawmeaiksovlon . tPSrhneeesdbsei,cnoAomrmi,easlG,Wdip.spt.raib2nu4dt0io-Cn2o.4c1h.rSatna,stWc.aGl.M(e1t9h67o)d.s,V6atrhiaEdnicteiotne,stlfoowrahSotmaotgeeUnneiivteyrsoifty 7. BSiookmale.trRy.R.W.aH.ndFRreoehmaFn.J.an(d16C9o).., SaBanrtFlreatntcitsecsot,opfp.ho3m7o0g.e3n7e1ity of variances. 8 SMneetdheocdosr.,6tGhWE.ditainodn,CloocwharaSnt,atWe.GU.niv(e1r9s6i7t)y. PrAensasl,ysAimseosf,Vappr.ia2n5c8e..275St,atistical 9 tDruenantemtetn,tCsWwi.th(1a9c5o5n)t.rolA.muJt. iApmleer.coSmtpata.rAisssooncp.r5o0c:e1d0u9re6-f1o1r2c9omparing several 10. WS.oHka.l.FrReRe.maanndanRodniCto,.,F.SJ.an(1F96r9a)n.cisKcrou,skpap.l-W3a8l8l-i3s8T9est. Biometry, 1. 6D(u3n)n:,2401..J2.52(.1964). Multiple comparisons using rank sums. Technometrics 12. SMiceGgreal,w-.Hil(1,95N6)e.w YNoornkp,aprpa.me9t6r-i1c04S.tatistics for the Behavioral Sciences, GO0G88 PROTOCOLAPPROVAL: FOR THE TESTING FACILITY le. -- George E. Dearlove, Ph.D., DABT Associate Director of Research LA Study Director Raymom G. York, Ph.D. Assduiate Director of Res DABT h bara J. Patters BA. hairperson, Institutional Animal Care and * Use Committee 418-008:PAGE F-27 Protocol 418.008 Page 27 2 sep iy Date 2M gf Date 2Mab Date FOR THE SPONSOR Dn [Ga SMtaurvdiynMTo.niCtaosre, D.V.M., Ph.D. 22 Mey Jae Date 0600689 418-008:PAGE F-28 ATTACHMENT 1 STUDY SCHEDULE 000650 ATTACHMENT 1 418-008PAGEF-29 ProtocPoalg4e181-00028 SCHEDULE 12 MAY 98 26 MAY 98 26 MAY 98- 15 JUL 98 26 MAY 98-19 AUG 98 09 JUN 98 - 22 JUN 98 2292JJUUNNSSBBPPMM--0269JJUULNISBBAAMM 2036.JJUUNL 988 204uL 98 A(FnoimgaelneRreacteiiopntr-atAsc).ciimation Begins RStaatrsto(f28DodasyasgebePfeorrieocdoh-aFboitGaetnieornaatnidon Male p`ceornitoidnuuinntgiltsharcoruifgihceaa1ft4e-rdsauycccoehsasbfiutlatmiaotning has been determined). DCaoessaagreeaPne-rSieocdti-oFneimnagl(e28RadtasyAssbseifgonreed to pcorheasbuitmaetdiognesatnatdiocno)n.tinuing through day 09 of NDaotsuaragle DPeelriivoedry- [F2e8mdaalyesRbaetfsorAesscoihganbeidtattoion tthhartodugohndoatyde2l4ivoefraprleitstuerm) eodr dgaesyt2at0ion (rats Postpartum (rats that deiiver a litten)} Dosage Period Estrous Cycle Evaluation. CoMhaalbeit1at(i0o7ndPaeyrsi)od (Maximum of 14 days). Male 2 (07 days) LFiarsstt PPoossssiibbllee DDaayy 00ooff PPrreessuummeedd GGeessttaattiioonn.. CFoompGleneetriaotnioofntMhaelCeohRaabtistaStaicorinfPiceerdioadft(eErarliest possible date). 2 The study initiation date is the day the Study Director signs the protocol, 600691 ATTACHMENT 1 418-008:PAGE F-30 ProtocPola4g1e8.2000182 03JUL 88 16.JUL 98 144UL98 31JUL 98 18.JUL 98 310uL98 03 AUG 98 20AUG 98 04 AUG 98 19 OCT 98- 02 NOV 98 16 NOV 98 10 NOV 98 - 27 NOV 98 30 NOV 98 - 17 DEC 98 06 APR 99 First Possible Day 10 of Presumed Gestation Caesarean-sectioning. Last Possible Day 10 of Presumed Gestation Caesarean-sectioning, First Possible Delivery (Day 21 of presumed gLeassttatPioosns)i.ble Delivery (Day 25 of presumed gestation). First Possible Day 25of Presumed Gestation Female Sacrifice. Last Possible Day 25 of Presumed Gestation Female Sacrifice. First Possible Day 21 Weaning (Dams and F1 generation pups not selected for continued observation sacrificed). Last Possible Day 21 Weaning. F1 Generation Postweaning Observations Begin (Details of tests cited in protocol), F1 Generation Cohabitation Period (Initiated when rats are approximately 90 days of age approximate initial date). F1 Generation Male Rats Sacrificed after Completion of Cohabitation Period Approximate Earliest Possible Date. Delivery Period - F1 Generation Dams/F2 Generation Litters (Approximate dates). Sacrifice of F1 Generation Dams and F2 `Generation Litters on Day 21 Postpartum (Approximate dates) Draft Final Report. 000692 418-008:PAGE F-31 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 000693 418-008:PAGE F-32 MDAATTEARISAHLEETSAFETY 31m Center sSts.144P-au1l0,00Minnesota 1-800-364-3877 or (612) 757-6501 (24 hours) AiCLonLpfyorrriaigaghthtit,sonr1fe9os9r8e,rvtehMdei.nnpeusCropootpsayeinMogifniapnnrgdo/poaernrdldyoMwaunntluioflaaidczitinungrginoMfg tCphorimospdauncy.ts 11)5 ptarhlieloorwienadfgorrpearesomtveiinodtnediissthaoGtbo:tpaiiendedinfrfoumll,withangno changes unless 2) Gne1i8tthreirbuttehde wciotphy tnhoer ithnetenotriiogninaolf eiasrnriensgolda oprrofoitthertwhiesreeon. TDRIAVDIESIONNA:ME: 3M CHEMICALS 109F8CN--U09M25B0E7FR-L0/U1UO0.R3PA-.7DC'.:Bra0n0d-51F1l3u5o-r0o9c0h5e4s-i1cal98S.u0rf2a0c7t-a0n1t04-5 00-51135-09085.5 ISS29U86E-D00:2010J1a2-n01u80a8r48y-1529,001-95018135-L093L62.7 8.0211-9916.1 00.51195. 039175 DSOUCPUEMRESNETD:ES:10N-o3v7em3b6e.r. 05, 1967 1. INGREDIENT PPOOTTAASSSSIIUUMM POTASSIUM PPPEEERRRFFFLLLUUUOOORRROOOAAALLLKKKYYYLLLSUSSLUUFLLFOFOONNNAAATTTEEE............!|. PPOOTTAASSSSIIUUMM PPEERRFFLLUUOORROOAALLKKYYLL SSUULLFOFNOANTEA.T.E....|| 2. PHYSICAL DATA CAS. HO PERCENT 32278594572-031.9.4-939.539.63382."87gs 36807227.02.68.51.5 3 Cs 3 vVBaAOpPIoOLrRIoPNeRGNEsSPOISITUNYRT:E::..1.1..1.11.11..1.1.1..1.1..1. yN/aA Wa SSEPOVELACUPIBOFIRILACITTIYGORNAR1VAITWTEaY:T:E.1R.:.11111111I111111T11I] jSiagne ca. "0.6 Waterss PERCENT VOLATILE:.............. 0(%Bulk) VIHSeCCTOISNIGTYp:oinot: iIlIIT I(D0.1% No Aqueous) APPLEiAgnRtANcCoEloArNeDd,ODOfRr:ee flowing powder. AOBreviations: NID - Not Deterained N/A - Not Applicable GA - Approximately (00694 JMaSnDSu:aryFC-299,5 F1L9U98ORAD Brand Fluorochemical Surfactant 3. FIRE AND EXPLOSION HAZARD DATA 418-008:PAGE F-33 PAGE 2 FFLLAASMHMABPLOIENTL:I.M.I.T.S..-..L.E.L.:.....1.1..1.0. NN/oAne AFULTAOMIMGANBILTEIOLNIMITTESMPE- RUAETLU:R.E.:.........]. NN//AA EXWTaItNeGrU,ISHCIarNbGonMEDdIiAo:xide, Dry chemical, Foam SPHpEeoCasIriAtLifvuFelIlREpprreFosItsGueHrcTetIiNvoGerPRcprOleoCstEshDuiUrnRegE',S:deimnacnldudibnrgeathheilmnegt,appsaerlaft-ucso,ntabiunnekde,r coat apnrdotepcatnitsv,e cboavnedrsinagroufnodr aerxmpso,sedwaiasrteasandofletghse, hefaadc.e wask, and UNUSSeeUALHazFaIrRdEouAsNDDeEcXoPmLpOoSsIOiNtioHnAZAsReDcSt:ion for products of combustion. 4. REACTIVITY DATA STABILITY: Stable INNCoOtMPAapTpIlBiIcLaIbTlYe.- MATERIALS/CONDITIONS TO AVOID: HAZARDOUS POLYMERIZATION: Hazardous polymerization Will not cour. HACZaArRbDoOnUSMoDnEoCxOiMdPeOSIaTndIONCarPbROoDnUCDTiSo:xide, Oxides of Sulfur, Fluoride, Toxic Vapors, Gases or Particulates. Hydrogen 5. ENVIROWMENTAL INFORMATION - SPOCILboLsmeproRvuESenPdOpNorSreEc:aautteironstofraovomidotdhuesrtisnegc.tioCnAsU.TIONVIacuAumv,acuuusme Wcelteansewreepcionugld abpepraonveidgnimettiaoln scoounrtcaei.ner.CleaSnealupthreesciodnuteaiwnietrh. water. Place in an REDCoOMnMoEtNDErDeleDaIsSePOStAoL:Waterways processes that could result oinr asqeuweart.ic Dcooncneotntruasteioinns pgrroedautcetrs otrhan mi1an/td1e0ursitoarfli.atlheoCrolmobcwuoesmsttmieorEncCiSa0plroodrfuaccLtiOslSOitwyiclolninceinnttchrleautdpieroensH.eF.nceIDniocsfipnoassraacltoembuinstiabnle alternative: Dispose of waste product in a facility permitted to Avbreviations: WID - Not Determined WIA - Not Applicasle GA - Approxim(a0teGOlSy - JHaDnSu:aryFG2-93,5 1FL9U%GRAD Brand Fluorochemical Surfactant 418-008:PAGE F-34 Pace 3 5. ENVIRONMENTAL INFORMATION (continued) accept chemical waste. ENB9Vl6Iu-RHeOrgM.iElNlATqASuLuantfiDicAsThAF(:iLsehpoLnCiSsO, maFcartohcehaidruMsi)n=n6o3w(Pmigs/Le,phaRlaessnbopnroTmreolusss)s=o3isnsagli, 3ga;iranBeorbsz)1+1 NmEgL/L; 4B-nr. 030, Diphnia Magna = 50 ee/1; enerss REVVGoOUlCLaAtLTieOlsResY HOIrNogFaOn&RiMcAETxICeOaoNpm:tpouSnodlsv:entNs/:A. N/A. EbSieanfcoerHeazrdeaigrsudplooaustsai)lo.ns vU.aSr.y,EPcAonsMualztardaopupsliHcaasbtlee Mreogsuhlearti=onsNovoer a(uhterhorurist.ies TThSiCs, p"rEoIdNuEcCtS, cCoOmSpLl,iesALCwSi,thMItThEe acnhdemKiocraela.registration requirements of EPFICRREA HHAAZZAARRDD: CLNAoSS:PRESSURE: No REACTIVITY: Mo AGUTE: Yes CHRONIC: Yes "6. SUGGESTED FIRST AID evmeIiancmuoetndetisa.actte:lGyetFliumsmhedieaytees wmeidtihcallargaetteanmtoiuonnt:s of water for at least 15 SkcITonanntecadominiatntaaectl:yedlculsohthisnkgi.n wItithirrliatragteioanmoupnetrssisotfs.watceerl.l Contaminated clothing berore reese. aRepmuoyveeician. Wash IHSIAitLgAnsTsiIgsOnyNsa:spytmopntsomcsontoicncuuer,, craelslovea ppheyrssiocniatno. fresh air. If IFDrSiHnakLLotwwEoD:glasses of water. Call a physician. 7. PRECAUTIONARY INFORMATION - evAevoriodTeeCyTeIOcNo:ntact. Wear vented goggles. Abbreviations: NID - Not Determined Nia - Not Appiicabie Gh | Approximately 000696 418-008:PAGE F-35 JMaSnDSu:aryFG2-99,5 1F9L9U8GRAD Brand Fluorochemical Surfactant PAGE 4 7. PRECAUTIONARY INFORMATION (continued) SKAIvNoiPdROTskEiCnTIOcNo:ntact. material. A pair of Wgleoavresapspardoeprfiraotem tghleovefsollwohweinnghamnadtleirnigalt(hsi)s are pcroeevrcesoromimnneagnl,depdcr:oovteercbatulitloysnl. irtuebPmbresort.easctiUnvseeecesogsanarermyeonrttsomarp(eroetvhoeferntthtehsaknifnogllclooovnwetisan)cgt:shouhtesad bpeolymeatdheylOefnse/iptohleyrvionfyltihdeenfeollcohwlionrgidemat(eSrairaalnse:x). RECUvsOeenMtMiEwlNiaDttEheDdaVpEapNrreToaIp.LrAiTaItPOerNo:vliodcealsuefxfhiacuisetntvevnetniltaitliaotni.on tUosemainintaaiwnell- iesminsostionasdeqbuealtoew, ruesceomampenpdreodprieaxtpeosurreespilirmaittosr.y prIofteecxthiaouns.t ventilation REASrvPeosIipRdiArTaObtRroYerastPhRibOanTsgEeCdToIfOoNnd:usati.rborSneelecctoncoennetroaftitohne fofollcoownitnagminNaInOtSsH aanpdproinved ahaclcfo-rmdaasnkcesuWpiptlhieOdSHAairrergeuslpaitriaotnosr:, full-face supplied air respirator. fhualllf--fmaacsek dduusstt aanndd mmiisstt rreessppiirraattoorr., PR0aE0rVeEaNnsTotIOtNheaotOr,FougdAhrCliCynIkDWEiNotTrhALsmsooIkaNepGESwanThdIeOnNw:autseirn.g tHhaisshprhoadnudcst.aftWearshhaenxdploisnegd and before eating. RECKOeeMpMENcDoEnDtaiSTnOeRrAGdEr:y. Keep container closed when not in use. FINRoEnfAlNaDsaEaXbPlLeO.SION AVOIDANCE: OTNcHoEoRntsamPmoRikEinCnaAgtU:iToInSOmNAoofRkYintghIeNWFhtOioRlbMeAaTcIcuOosNi:nagnd/tohris smporkoeducatnd calneadretsoulttheinformation tOhfistheMSDhSa.zardous decomposition products mentioned in section of HMIS HAZARD RATINGS: WPEEARLSTOHN:AL2PROFTLEACMTMIAOBNI:LITXY:(Se0e RpErAeCcTaIuVtIiToYn:s, 0section 7.) EXPOSURE LIMITS INGREDIENT VALUE unIT PPOOTTAASSSSIIUUMM POTASSIUM PPEERRFFLLUUOORROOAALLKKYYLL PERFLUROALKYL SSUULLFFOONNAATTEE....!. SULFONATE | 00..11 0.1 MMGG//MM33 Wa/ma POTASSIUM PERFLUCROALKYL SULFONATE... 0.1 mWa/ma Abbreviations: N/D - Not Determined N/A - Not Applicable TYPE AUTH SKIN TTWuAa aatvv TTwwaa n mv v CA - Approximately 000697 MSDS: FC-85 January 29, FLUORAD 1998 Brand Fluorochemical Surfactant 8. HEALTH HAZARD DATA (continued) 418-008:PAGE F-37 PAGE REPNRoOtDUtCeTrIaVtEo/gDeEniVcELOiPnHEtNheTALratTOXaItNSo:ral doses belo maternally toxic Levels. OTTHhEiRsHEpArLoTduHctHAZisARnDotINkFnOoRwMnATItOoN:contain California Proposition 65. any substances regulated under A Product Toxicity Summary Sheet is available. SECTION CHANGE DATES HEADING SECTION CHANGED SINCE November 05, 1997 ISSUE Apbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately TbIehMePLcIoiErnDrf,eocrtmIaNatCsiLUoDnoIfNiGnt,hetBhUdiTsateNMOaTtisesrLuiIeaMdlI.TESDa3fMeTOt,yMAAKDNEaYStaNIOSMhPeLWeIAtERDRA(NWMTASIRDESRS)A,NTiYsEXOPbFReElSiSeEvDedORto wMPhEEeRRtFChOHeRArMNATNAtCBhEeILIO3RMTYpUrSOoRAdGuEFcItOTFNEisSTSRAfDiFEtO.RfAorUsPeaArRpTaIirCstUiLrcAeuRslpaoPrnUsRipPbuOlrSepEosfOeRoraCnOddUeRtSesErumiitOnaFibnlge for ucuasnneirq'uaseflfyemcetwtihttohhdeinofutsehueseaundsoerra'paspplplikicncaoatwtilioeondng.eofanGadiv3ecMnonptrtrohodelu,cvta,ritiestoiymseeosoffsefnwathciitacolhrsatrhteahtat tphaertiucsuerlarevpaulruaptoese tahned 3sMuiptraobdluectfotro udseetre'rsminmeetwhohdethoferusiet iosr faiptplifcoartiaon. QJonuueeprtrrooovritsdh,eesormeiimnsoftsoeiromnapstoisoosrnibaiillntieterylaetctithorantosnieclinecftotrrhosisniacsinfatorrasmneasrtfvieiorcne,mat3yoMihmatavsekecsursetnsooumletresd. i[inneffpoorrreamsaaettniitooanntiooinnbstathiaenseMdtSoDfSriotmasvaaciolmdapabltleaetbaesndeeisrsemcaytolrynoatfcrcoubrmeacaSyMs.. curIrnenatddaistiotnh,e 000699 418-008:PAGE F-36 MSDS: FC-95 January 29, FLUORAD 1998 Brand Fluorochemical Surfactant PAGE 5 EXPOSURE LIMITS (continued) INGREDIENT VALUE UNIT TYPE AUTH SKIN POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 TM am v t=heSKIpNoteNnOtTiAaTlIONc:ontrLiisbtuetdionsubtsotatnhceesoviernadlilcateexdposwuirteh b'yY' thuendecrutaSnKeIoNusrefreoruteto biynclduidriencgt cmuocnotuasctmweimtbhranteheansdubsetyae,ncee.itheVrehibcyleasirbcoarnnealotre,r sWkoirne apbasrotripctuiloanr.ly, S OU3RM:CE OFaMEXRPeOcSUoRmEmenLdIeMdITEDxApToAs:ure Guidelines 8. HEALTH HAZARD DATA EYEMilCdONTEAyCeT:Irritation: signs/symptoms can include redness, swelling, pain, and tearing. SKIN CONTACT: Msiilgdns/SskyinsptIormrsitcaatnioninc(laufdteerrepdrnoelsosn,gedsweolrlirnegp,eatanedd ictocnhtiancgt.): Meaxytenbdeedabstoinreb.ed through the skin and persist in the body for an INMHaAyLATbIeONh:armful if inhaled. TMiamye be absorbed by inhalation and persist in the body for an extended Single overexposure, above recommended guidelines, may cause: sIorrreinteastsionof (tuhpepernosreespaindrattohrryo)a:t, sicgonusg/hsinygsptaonmds scnaeneziinngc.lude IF ISnWgAeLsLtOiWoEnD:is not a likely route of exposure to this product. Illness may result this material. from a single swallowing of a moderate quantity of May be harmful if swallowed. MUTAGENICITY: Mutagenicity assays indicate the product is not mutagenic. Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately (00698 418-008:PAGE F-38 ATTACHMENT 3 TEST ARTICLE PREPARATION PROCEDURE 600700 ` 418-008:PAGE F-39 ATTACHMENT 3 Version: 418.P0r0o8to(c2o1lM4A18Y-0g0e8 TEST ARTICLE PREPARATION PROCEDURE Page 1013 Test Article: PFOS. Vehicle: 0.5% Tween 80 in R.O. Deionized Water. A. Purpose: TofhdeopsuargpeosseuosfptehnissiopnrsoocfedPuFreOSis taondprtohveidceonatrmoeltahrtoidclfeofrotrhoeraplreparation `administration to rats on Argus Study 418-008. B. General Information: 1. sAlplecsiufsyptehnesipornotcoocnotlaniunemrbserwi,lltebset alratbieclleedidaenndtifcioclaotriocno,dAerdg.usEbaacthchlabel will number, concentration, and storage conditions. dosage level, preparation date, expiration date 2. Suspensions wil be prepared: X_ Day -- Weeky _For__daysofuse 3. Suspensions will be prepared at a final dosage volume of5mLikg. 4. safety: XX__ GDulsotv-eMsi,sltabRecsopaitr,atgoorggles or safety glasses and faceshield -- Half-Face FulFFace Respirator Respirator/Positive Pressure Hood Tyvek Suit/Apron 5. --DosageYessolutions adjuXs_ted Nfoor F(rCaelecublaasteioannsdba%sePduroitny.100%) __ FreeBase __ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol. 000701 418-008:PAGE F40 ATTACHMENT 3 Version:416P.r0o0t8o(c2o1lM4A1Y8-0580)8 TEST ARTICLE PREPARATION PROCEDURE Page20f3 NOTE tTheestloarwtidcolseawgiell.beStpirrebpaarrseadreastoabseeraiadlddeidluttoiotnhfercoomnttahienehrisg;h mdioxsiangge to should occur during sampling and/or dosage administration. C. Preparation of the Control Group: 1. AAdRdTItCheLEreCqAuiLrCedULaAmoTuInOtNoSf fvoerhiecxlaecttoquaanntaiptpireso.p)riate vessel, (See TEST D. Test Article Solution Preparation: 1. iTnotporaenpaaprperotphreia0t.e6l4y-msgi/zmedL,.,la(bgerloeudpcVo)ntsaoilnuetri.on, add 160 mg of test article 2. Q.5.t0.250 mL with the vehicle and mix by inversion, 3. `Aadpdpraoxsiimratbealrya3n0dmhienauttetshe(oprreupntairlatthieontetsot8a0rticCleihnaaswdaitsesrolbveadt)h.for 4 tRheemsoovluetitohneesqouliuitbiroantfersotmotrhoeowmatteermpbeartaht,uraen.d slowly spin over night while 5. sTooluptrieonpatroeagnraopupproIpV,ri(attheely0.l3a2be-lmegdimcoLntsaoilunteiro,n)t,heanddq.51.00tom2L00ofmgLrowuipthV vehicle. Add a sti bar and mix until uniform. 6 sToolpurtieopnatroeagnroauppprollplri(attheel0y.l0a8b-emlegd/mcLonstoaliunteiro,n)t,haenddq.55.0 tmoL20o0f gmrLouwpitIhV vehicle. Add a stirbar and mix until uniform 6006702 418-008:PAGE F-41 ATTACHMENT 3 Versio: 418.0Pr0o8t,o2cP1oalMg4Ae1Y83.900180318 TEST ARTICLE PREPARATION PROCEDURE r TsooluptrieopnatroeagnroauppproIlp, r(itahtee0ly.0l2ab-emlge/dmcLonstoaliutnieorn,),thaedndq.550. mtoL2o0f0gmrLo.upwiItlh vehicle. Add a stir bar and mix until uniform. Witten by: lr , A noproved ty ames) OF he. vate: 2b 96 U uc ake) Clarification: X No _Y Yes (See attached clarification form.) InitialDate: MC |-Il-gq 600703 418-008:PAGE F-42 angus : TEST APRRTOICCELDEU/RSETRCSLTAARNICFEICPARTEIPOANRATION Protecol:_4/-cog Verston:_4/5-005 (31 omar DCalcaerifoifcation shijue EC ip PrSeptaerpascion ectr Clartficacion --222020atat ooff VVeheichleiisodedeilleedd rsie prporain.g, Cootre] Cocup 8 zlshe -_ --- E -_-- la 5 Divecs: = -_ EE e a, - -- T-_T-- SEE bee mm -_-- -_-- Ee - - TE T mme hm -_-- Ee -_-- -_-- TT sh bm hm Revieve[ d byS : SMCE . CE er lossy D0a8t.e1:557_T_a#s-e1R5E--901-07 000704 418-008:PAGE F-43 -- A ---- iu scna -- cs rtes ------ArgHu-- os5rTE 0Rs5ehaSsaehmS ae,recPhehaynrRLinDasrbiyovlreava,atnBo8iruwialeeks1i,5nnsI6gn4cA4. ---- PROTOCOL 418-008 `COMBINED ORAL(GAVAGE) FERTILITY,DEVELOPMENTAL AND PERINATALUPOSTNATAL REPRODUCTION TOXICITY STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: 6295.9 ---- Amendment 1 - June 2, 1998 1. Storage (page 3 of the protocol) t[EefmfpeecrtiavteurDeatoev:ernMiagyht2,6r,at1h9e9r8]thaPnrefproazreend. formulations will be stored at room ReaforsChoannge pTrheipsarcahtaionng.e was made to clarify the protocol and facilitate dose formulation 2. FreqoufPerepnarcatiyon (page 4 of the protocol) [Effective Date: May 26, 1998] The vehicle (0.5% Tween 80 in R.O. Deionized Water) will be prepared weekly by adding 15 mLs of Tween 80 to 2985 mL of R.O. Deionized Water. Reason for Change Tprheipsacrahtainogne was made to clarify the protocol and facilitate dose formulation 3. Stabilty (page 4 of the protocol: [Effective Date: May 27, 1998] The Sponsor has confirmed a 48-hourstability on the test article in 0.5% Tween 80 solutions. This allows for preparations to be made one day prior to the day of dose administration. Due to thelengthy 000705 418-008:PAGE F44 _ Prot"oAcmoeln4d18m-e0n01t8 Page2 preparation dosing. procedure, dose solutions may be made one day prior to the day of Rea forsCho angne: This change preparation. was made to clarify the protocol and faciltate dose formulation lo olin ans Hy CN DiArleacntMo.roHfobReersmeaanr,chPh.D., DABT Date ARsasyomciiantde GD.irYeoctrokr, oPfh.DR.esp[aDrAcBhT and Study Director 0zq00-95 Date 2lia Co; forked DMeemnbaeCr., LIenbstoi,tuVt.iMon.aDl.Animal Care Date and Use Committee 9 7Gor 8Jn tre SMtaurdvyinMTo.niCtaosre, DV.M., PhD. Date 000706 418-008:PAGE F-45 Argiis prey neseancy Argus Research Laboratories, Inc. --e--e----------p--S ----Es----------------H--orTs--ahrasm--,sPee--rnnis--yoFl.v3a--n1i9a--44153--05484 PROTOCOL 418-008 COMBIANNEDDPOERRAILNA(TGAAUVAPGOES)TNFAERTTAILLIRTEY,PRDOEDVUECLTOIPOMNENTAL TOXICITY STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: 6295.9 Amendment 2 - June 11, 1998 ---- e -- ee 1. Body Weights - Female Rats (page 12of the protocol): a(ElfsfoecbteivceolDlaetcet:edMoanyF2o6,ge1n9e9r8a]tioBnodfyemwaeliegshtosnapnodstfpeaerdtcumondsauymp1t0.ion values wil ReafosrChoangne: This information was added F1 generation females. to the protocol to match data collection on 2. Scheduled Sacrifice of Male Rats (page 14 of the protocal): [Effective Date: will be weighed May 26, 1998] individually At scheduled sacrifice of Fo male rats, all organs Reason for Change: This change clarifies the protocol, 3. S(pcahgeedu1l5edofStahceripfriocteoc-oFle):male Rats Assigned to Caesarean-Sectioning [Effective Date: May 26, 1998] Uteri of non-pregnant rats will not be retained. 600707 418-008:PAGE F46 Pro`toAcmolen41d8m.e0n02:8 Page2 Rea forsCho angn e: This change was made at the request of the Sponsor. 4. Scheduled Sacrifice - 1 Generation Male Rats (page 20 of the protocol) [pErfofsetcattievse,Dwaitlel:bMeawyei2g6h,e1d9a9t8]scSheedmuilneadl vsaecsriicfliecse (owfitalhlaFn1dgweintehroauttiofnluimdas)leanradts, ReaforsChoangne: gTehniesriantfioornmamtailoens.was added to the protocol to match data collection on Fo Leber ran UL Oth vse ADliraenctMo.r oHfobReersmeaanr,chPh.D., DABT Date ARsasyockioantde GD.irYeoctrokr, ohhoRls.eaDrAcBhT Date and Study Director Lomne LC dbnln tsier MDeemnbaerC,. LIenbstoi,tuVt.ioMn.aDl.Animal Ca/re Date and Use Committee Was Tie Manin T. Case, D.V.M. Ph2O. fatoDastee Study Monitor 000708 . 418-008:PAGE F-47 . eBra Argus30R5esSehaeracrhyLOabeor,atuoriels, yin4c. ----------------LAS--ORA--TIR--ES--------------------HorT--shaa--m,s--Pens--nsy--lvaa--nrieas--a19s--50s84y4] PROTOCOL 418.008 COMBAINNEDDPOERRAILN(AGTAAVUAPGOES)TNFEARTTAILLIRTYE,PRDOEDVUECLTOIPOMNENTAL TOXICITY STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: 6295.9 Amendment 3 - June 25, 1998 -_-- 1 AdminiMsetthord aantdFireqouennc,y (Page 10 of the protocol): c[aElffceucltaitvieonDatthee:dJousnaege19p,er1i9o9d8]wilDlubeeteoxatepnrdoegdrafomrmainngadedrirtoironinald2oswaegeeks and the ecrorhoarbiitnadtoiosnagpeerciaoldcuwillaltoiocncsurcatuwsoewdetehkesmlaalteer rtahtasntoorriegicneailvley aspcpherdouxliemda.telTyh8e9% to 90% 99% of tohfethteartgaertgeedteddosdaogsea.geThainsdofcecmuarlreedraftosr to receive sixof the saepvpernoxdiamyasteilnyt8he5% to second and third weeks and two daysof the fourth week of the dosage period. Reason for Change: This change cohabitation was made to ensure that all rats receive the correct dosage before 60U709 418-008:PAGE F-48 Prot`oAcmolen41d8m-e0n03t8 Page2 2. Schedule (Attachment 1ofthe protocol): 12 MAY 98 SCHEDULE Animal Receipt (Fo generation r-atAsc)c.limation Begins 26 MAY 98 26 MAY 98-29 JUL 88 26 MAY 98 - 02 SEP 98 09 JUN 98-06 JUL 88 0163JJUULL99BBPPMM--1230JJUULLI9BBAAMM 07JUL 98 20JUL 98 03AUG 98 17 JUL 98 30JUL 98 Start Rats (o4f2DdoasyasgebePfeorrieocdo-haFboitGaetnieonraatnidon Male `pceornitoidnuuinntigltsharcoruifgihceaa1ft4e-rdasuycccoehsasbfiutlatmiaotning has been determined). CDaoessaagreeaPne-rSieocdti-oFneimnagl(e42RadtasyAssbseifgonreed to pcorheasbuimtaetdiognesatnatdiocno)n.tinuing through day 09 of NDaotsuaragleDPeelriivoedry- [F4e2mdaalyesRbaetfsorAessciohganbeidtattoion through that do day 24 of not deliver presumed a litter) or gestation day 20 (rats postpartum (rats that delivear litter) Dosage Period Estrous Cycle Evaluation. Cohabitation Period Male 1 (07 days) (Maximum of 14 days). Ma2l(e07 days) FLiarsstt PPoossssiibbllee DDaayy 00 ooff PPrreessuummeedd GGeessttaattiioonn,. CFoomGpelneetriaotnioofntMhaelCeohRaabtistaStaicornifPiceerdioadft(eErarliest possible date). CFiaresstaProesasni-bsleectDiaoyni1n0g.of Presumed Gestation CLaaesstaProesasni-bsleectDiaoyni1n0g.of Presumed Gestation 660710 28JUL 98 14 AUG 98 01AUG 98 14AUG 98 17 AUG 98 03 SEP 98 18 AUG 98 02 NOV 98 - 16 NOV 98 30 NOV 98 24 NOV 98 - 11 DEC 88 14 DEC 98.- 31 DEC 98 06 APR 99 418-008:PAGE F-49 ProAtomceoln4d1m8e-0n0t38 Page 3 First Possible gestation). Delivery (Day 21 of presumed gLeassttatPioosns)i.ble Delivery (Day 25 of presumed FFiermstalPeosSsaicbrlieficDea.y 25 of Presumed Gestation FLaesmtalPeosSsaicbrliefiDcea.y 25 of Presumed Gestation FFi1rsgtePnoesrsaitbiloen Dpauyps21noWtesaenliecntged(Dfoarmcsonatnidnued oLbassetrPvoastsiiobnlesacDraifyic2e1d)W. eaning. BFe1gGienn(eDreattaiilosnofPotsetswtseacniitendg iOn bpsreortovcaotli)o. ns wF1heGnenreartsatairoenaCpophraobxiitmaattieolny P8e0ridoadys(Inoiftiaagteed approximate initial date). CFo1mpGleneetriaotnioofnCMoahlaebiRtaattisoSnacPreirfiiocded- after Approximate Earliest Possible Date. GDeelnievreartyioPenrLiiotdte-rsF1(AGpepnreorxaitmiaotne Ddaatmess)/.F2 SGaecnreirfaicteioonf LFi1ttGeresneornatDiaoyn2D1aPmosstapnadrtFu2m (Approximate dates). Draft Final 600711 418-008:PAGE F-50 Pro`toAcmoeln4d1m8e.n0t083 Page Rea forsCho angne: wTheiesksscahenddutlheewcaoshacbihtaantgioendpdeureiotdootchceurerxitnegnstiwoonowfetehkes dlaotseirntghapnersicohdedbuyltedw.o Qin -- 1S . ef 25s $.~ ADliraenctMo.r HofobReersmeaanr,chPh.D., DABT Date 25-Guw 55 ARsasyomcioantde GDi.rector-Yddrfk, RFhe.Ds.,eaDrAcBhT Date and Study Director Lena Chelr 25 esr Dena C. Lebo, V.M.D. Member, Institutional Animal Date Care and Use Commitee Mee TC 134 57 Marvin T. Case, Study Monitor DVM. PhD. Date C0072 PRIMEDICA _-- 418-008:PAGE F-51 Arg5u0s5ReS search b Laborsiois nc TelTeepehtoanxe:: G31139))43434.7851857 PROTOCOL 418-008 PERINACTOAMLBPIONSETDNAOTRAALLR(EGPARVAOGDEU)CTFIEROTNILTIOTXYI,CDIETYVESLTOUPDMYEONFTAPLFOASNDIN RATS SPONSOR'S STUDY NUMBER: 6295.9 Amendment 4 - July 14, 1998 _-- 1. TpersottoActoil)c:le Preparation Procedure (Version 29 JUN 98ofAttachment 3 to the p[Erfefpeacrtievde eDaatceh:daJyunwiell29b,e 1i9nc9r8e]asTedh.e vTohleumneeowfTtehsettAerstticalretiPcreepsaorluattiioonns Procedure is attached to this amendment Reafosr Choannge dItasybneeccaesussaeryofttohienicnrceraesaestihneg vboolduymweeiogfhtthseotfesttheartriactlse, solutions prepared each A= A Huy H/GAesdsiocgiiate. DDieraerctotvoerlof ResPhe.D.arDcAhBT Date faz A RAsasyocmioatdeGD.irYeoctro,r oPRfD.ReseDaArcBhT ry-guc-s, Date. Study Director Meson Tl 3g 95 BChaaribraprearsJ.onP,tInbbftsiotunt,ioBn.Aal Animal CaDraete and Use Committee: SMatruvdiynMTo.niCtaosre, D.V.M., Ph.D. Date Nn3 COU PRS - 418-008:PAGE F-52 ATTACHMENT 3 Version: 418-P0r0ot8o.c2o9l J4U1N8.0908)8 TEST ARTICLE PREPARATION PROCEDURE Page 1of3 Test Article: PFOS. Vehicle: 0.5% Tween 80 in R.O. Deionized Water. A. Purpose:ofThdeospuargpeosseuosfptehnissiopnrsocoefdPuFreOiSs atondprtohveidceonatrmoeltahrtoidclfeofrotrhoeraplreparation administration to rats on Argus Study 418-008. B. General Information: 1. sAlplecsiufsyptehnesipornotcoocnotlaniunmerbserwi,lltebset alratbieclleedidaenntdifcioclaotironc,odAerdg.usEbaacthchlabel will number, concentration, and storage conditions. dosage level, preparation date, expiration date 2. Suspensions _X_ Daily will be prepared: Weekly _For__daysofuse 3. Suspensions will be prepared ata final dosage volume of mLikg. 4. safety X_ X_ DGulsotv:eMsi,stlaRbecsopaitr,atgoorggles or safely glasses and faceshield _ -- Half-Face Full-Face Respirator Respirator/Positive Pressure Hood Tyvek SuitApron 5. --DosageYessolutions adjuXs_ted Nfoor F(Craelecublaasteioannsdba%sePduriotny.100%) -- FreeBase __ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited in protocol. GOoU'ERS 418-008:PAGE F-53 ATTACHMENT 3 Version: 418Pro-to0co0lJ84U1.N6.9208098) TEST ARTICLE PREPARATION PROCEDURE Page 2013 NOTE: Ttheestloarwtidcolseawgiell. beStpirrebpaarrseadreastoa bseeraiadlddeidluttoiotnhfercoomnttahienehrisg;h dosage mixing to `should occur during sampling and/or dosage administration. C. Preparationofthe Control Group: 1. AARdTdItChLeEreCquAiLrCedULamAoTuInOtNoSf fvoerhiecxlaecttoquaanntaiptpireos.p)riate vessel. (See TEST D. Test Atticle Solution Preparation: 1. Tinotoparnepaaprperotphreia0t.e6l4y-smigz/emdL,,la(bgerloeudpcVo)ntsaoilnuetri.on, add 224 mgoftest article 2. Q.S.t0.350 mL with the vehicle. 3. A`adpdpraoxsitmiratbealrya3n0dmhienauttetshe(oprreupntairlattihoentetsot8a0rticCleinhaaswdaitsesrolbvaetdh).for 4. eRqeumiloivbreattehsetsoorluotoimontfermopemrtahteurwea.ter bath, and spin while the solution 5. TsooluptrieopnatroeagnraopupproIpV,ri(attheel0y.l3a2be-lmegd/mcoLntsaoilunteiro,n)t,heandda.51.50tom3L00ofmgLrowuipthV vehicle. Add a stirbar and mix until uniform. 6. sTooluptrieonpatroeagnroauppprolplri(attheel0y.l0a8b-emlegd/mcoLnstoaliunteiro,n)t,heanddq.65.5tmoL26o0f gmrLouwpitIhV vehicle. Add a stir bar and mix until uniform. GOV KB 418-008:PAGE F-54 ATTACHMENT 3 Version: 418-P0r0o8to2co9lJ4U1N8.5080)8 TEST ARTICLE PREPARATION PROCEDURE Page 3of3 7. Witten by: TsooluptrieopnatroeagnroauppproI.pr(itahteel0y.0la2b-emlge/dmcLonstoaliutnieorn,),thaedndq.550. vehicle. Add a stir bar and mix until uniform. mtoL2o0f0gmroLupwiIith . < Approved by: Date: /3-Fua Clarification: 2" No __ Yes (See attached clarification form.) Inital/Date 2 A 200 99 GOUTIR OPRIMEDICA _-- 418-008:PAGE F-55 TTHaHorie EiP1AAI 55E03 Teche PROTOCOL 418-008 PERINACTAOLM/BPIONSETDNAOTRAALLR(EGPARVAOGDEU)CTFEIROTNILTIOTXYI,CDIETVYESLTOUPDMYEONFTAPLFOASNDIN RATS. `SPONSOR'S STUDY NUMBER: 6295.9 -_ Amendment 5 - July 17, 1998 1 Concentration Analyses (page 5 of the protocol): Samples last week (5 mL of the each) from F1 dosage each concentration will administration to verify be taken during the first concentrations of the and Prepared test article formulations. TRrisechafaons rgChwo aansgnem:ace because dosage aaminsaton was extended onl the F1 generation male and female rats. 2. Sex (page 6of the protocol): The F1 generation male and female pups will be given the test article or vehicle. Reason for Change: Tinhfeosrmeatcihoannagbeosuwtetrhee mefafdecetsaotfthteherteeqsutesartt of icl ethoenStphoensseocroinndgoerndeerrattoioprno,vide 3. Method and Frequency (page 10 of the protocol) ToFr2haelgleyFn1(egrgaaevtnaiegorena)tpiouonpnsdmawaiyllle1naoptnodsbtefweedmiaarnelicentlgPyutgphisrvoewunilglthhbteehteaedsdmaiynibsetfeorreedsatchreiftiecse.t article meaxtpeorsneadl tmoiltkhedutreisntgartthieclleadctuartiinogn mpaertieordn.al gestationt (airntiucltee,robuetxpmoasyurbee) poorsvsiiably I? oo 418-008:PAGE F-56 Pro`tAocmoeln4c1m8e-n0t058 Page2 Rea fos rCho angne: oTfheexspeocshuarengteosFwoegreenemraatdieon.at the request of the Sponsor and follow the method 4. DC oso agenLec vele s, ntrataindo Volnums es (page 11 of theprotocol): r Nummcer | E52 | cpoRematmseern | Sossze --| Conrcaammrsa)ten | (ming)| Argus Baen Numer Lo J Pte TeTn FeeToToTon en] Jeworcar] LoTeToTT on ToTensronmenn] Reason for Change: tThheisFc1hgaenngeerawtaiosnmmaadlee banedcafuesmealdeosraatgs.e administration was extended to include 5 Tests. Analyses and Measurements - F1 Generation (page 16 of the protocol): Clinical Observations andlor General Appearance: Preweaning Period: Once daily. Dosage Period: Maternal Behavior. aTwnidcoendcaeilay.ppPrroixoirmtaotedloysaognee hadomuirnipsotsrtadtoisoange. oDbasyesrv1e, d4,ab7no.r1m4alanbdeh2a1vipoorstwiplalrbtuemr.ecAonrdyed daily coords 418-008:PAGE F-57 ProtAomceonld4m1e8.n0t08 Page3 BWodey igh-t Malse: Preweaning Period: Dosage Period: Days 1 (birth), 4, 7, 14 and 21 postpartum Weekly. Sacrifice: Terminal weight. BWodyeigh-Fetmalse: Preweaning Period: Dosage Period: Sacrifice: Days 1 (birth), 4, 7, 14 and 21 postpartum. gWeesetkaltiyotnoacnodhaobintaDtaiyosn.1,D4a,il7y adnurdin1g4 presumed postpartum (rats assigned to natural delivery). `Terminal weigh. ReafosrChoangne: TinhcelsuedecthhaenFg1esgewneerreatmiaondemableecaaunsdefdeomaslaegeraatdsministration was extended to 6. EA/E2 Generation protocol): Pups Not Selected for Continued Observation (page 22ofthe `OcSunlalmdepdalyepsu4pwpislolsftbrpoeamrcotGlurlmoe,ucptteshdeIf,srItoomamnaadlcl Vhpuc(poorsnthfeirngothmsesf(tomuidlroktsocaufgriedv)egworifloltuhbpeealvcaaorilglleaesbcltteel)id.tefrrsomin ctionhltelosepcetoitlohynrp.eresoapdymolpselanegesetwugiblrleousbpeas.nsdhiIfpnrdpoiezvdiedn(ufaarlto-zp2eu0npoCs.namdArpfylteeiscre)cwioltmlopbKlreeitscioJo.mnbHoiafnnsesademnbp,ylePlhi.tDer. at r3BeuMciilpEdinienvngitr2ao-nn3dmE-et0nh9te,a,lStSTtu.edcyPhanMuoolnl,ioMtgioynranwneisdlolStbaeafen5ot5ty1if3Si3eer-dv3ii3nc3ea1sdf,vo9ar3na5cneaBluoyfssihssa.AmvpBelonetuhes,htihpement. 000734 418-008:PAGE F-58 Pro"toAcmoeln4d1m8e.n0t0s8 Pages ReaforsCho angne: tCeosltleacrttiicolneoifs mrielakchsianmgptlheespwueprsevrieaqluaecsttateidonby the Sponsor to determineifthe ~rGekorbperE.eDcearlA oveP/h.D. DAB7T34D-a9te Associate Director oF Research B RaymbndG. York\oa h.0, DABT Date Associate Directorof Research Study Director Ce Chitin fe mii CBhaaribraprearsJ.onP,aItnsetirtsutoionnalB.AA. nimal CaDraete and Use Committee pss TC 200545 SMatruvdiynMTo.niCtaosre, D.V.M., PhD Date 72 (00% OPRIMEDICA . 418-008:PAGE F-59 ArtorTsmateepibaGaibRrA rr,earntc PROTOCOL 418-008 PERINA`TCAOLM/BPIONSETDNOATRAALL R(EGAPVRAOGDEU)CFTEIROTNILTITOYX,ICDIETVYESLTOUPDMYENOTFAPLFAOSNDIN RATS SPONSOR'S STUDY NUMBER: 6295.9 -- tSAt `Amendment 6 - 13 August 1998 1 Necropsy (page 20oftheprotocol): On postpartum day 21, litters will be sacrificed. the 1.6 mg/kg/day dosage group (Group IV) dams and ReaforsChoangne: GlarcotautpioInV(wmoarstatlietrymiannadterdedatucweedanbiondgy bweecigahutsseaonfdthdeelsaeyveedrdeevpeulpoptomxeincitt)y,during 2 Male and Female Rats Assigned ofthe protocol): to Pharmacokinetic Sample Collection (page 14 Reasan for Change On postpartum Day 21, the livers remaining groups will be excised, of the Pups pooled per from five litters in each of the until shipment to theSponsor. litter, frozen and retained at-70C This change was made at the request of the Sponsor for possible analysis. 3. Schedule rifice -Fe le Ra No Surviving Pups (page 15 of theprotocol): il live ind Dams with Ovaries will be retained in neutral buffered 10% formalin. 2\ 000236 418-008:PAGE F-60 AmendmPeangtes2 Rea forsCho angn e: `This change was made to clarify the protoc [$6 eorgf E. Associate Dearlove, Ph.D., DABT DirectorofResearch Date LPs 98 nd G. York, 7.0. DABT Date ; AStsugsyociaDitreecDtiorrector of Redearch g 8 CL fdbic asd Tene Fd ov P SCahaairrpe)rson,atIntsetristounti,oRiaBlJA.Animal CaDraete MStaurdvyiMTno.niCtaosre, D.V.M., Ph.D. Date and Use Committee 722 0002 P".RIMEDICA -_-- 418-008:PAGE F-61 Acgy50sssRSesheaerychpLaoboeratB oruiiess , Inc TelTehpehtoanxe:: (O21155)) 4433.843981)8 TT PROTOCOL 418-008. PERINACTOAMLBPIONSETDNAOTRAALL R(EGPAVRAOGDEU)CFTEIROTNILTITOYX,ICDIETVYESLTOUPDMYEONTFAPLFAOSNDIN RATS SPONSOR'S STUDY NUMBER: 6295.9 _-- Amendment 7 - 24 November 1998 1. Natural Delivery (page 13ofthe protocol) oTfhteheLefnirgstt hpoufp Pdairvtiudreidtibony (Nt-i1mepoufpsdeilniveearcyhofliattsert) pwuilpnmoitnbues ctahlecutliamteeod.fdelivery Reafos rCho angne: A litter watch was not required by the Sponsor. A 2 bee Zone 7] H. 2usorsg BAsasrocgieat&e DDiareacrtoovroe,f PRensea.rDchABT Date Ra Associate GD.irYeocrtko,r PhS. of DABT Resdarch Date Study Director Dm Clin ahr CDheaniarpCe.rsLoenb,o,InVst.iMt.uDt.ional Animal CaDraete and Use Committee Pip TE. 25pp SMtaurdviynMTo.niCtaosre, D.V.M., Ph.D. Date 006738 PRIMEDICA _-- 418-008:PAGE F-62 ArJ emasEuhLaeaEbArs3a,50 ese PROTOCOL 418-008 PERINACTOAML/BPIONSETDNAOTRAALL R(EGPAVRAOGDEU)CFTEIROTNILTITOYX,ICDIETVYESLTOUPDMYEONFTAPLFAOSNDIN RATS `SPONSOR'S STUDY NUMBER: 6295.9 -_ `Amendment 8 - 18 May 1999 4. Concentration Analyses (page 5 of theprotocol): TTceohsnetdiucnocgntcFeaedcnitalirttaytt.hieondissacrmeptlieons owfertheesSepnotntsootrheanSdponnosorre.porStawmilpllbeeasneanltysteo sthweill be Reason for hange: This change was made at the request of the Sponsor to clarify the protocol. 2. MofaltheeapnrdotoFceoml)a:le Rats Assigned to Pharmacokinetic Sample Collection (page 14 The liver and serum `samples were sent to theSponsor. analyzed at the discretion of theSponsor. Samples will be Reason forChange: This change was made at the requestof the Sponsor to clarify the protocol. 3. `EA/mFe2ndGmeennetra5tioofnthPeupprostoNcootl)S:elected for Continued Observation (page 22 and STthoemsatcohmacocnhtecnotnstewniltlsobfe tahneasleyzpeudpsatwtehreedissecnrtettiootnohfe `tShpeonSspoornsfoorra.nalysis. JM 600739 418-008PAGE F-63 Reason for Change: ProtAomceolnd41m6e.n0t0s8 Page? This change was made at the request of the Sponsor to clarify the protocol. Lm orgh E. Dearlove, Ph.D. DABT Date Associate Director of Research 02 J Ra<yymoornde G. York, Kn.0) DABT Associate Director of Research Study Director sein Bate qJi come] Windond Lemnyts `ena C. Lebo, V.M.D. Date Chairperson, Institutional Animal Care and Use Committee Mts: Tse img zr. Marvin T. Case, D.V. M., Ph.D. Study Monitor Date 215 000740 APPENDIX G DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 3% GO0EL 418-008:PAGE G-1 DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY accurately evaluate this parameter. 1 On 8 August 1998, the 5 mL. concentration sample was not taken during the first weekof dosage administration for the F1 generation rats. This deviation did not adversely affect the outcome or interpretation of the study because sufficient data were collected to 2. On 3 to 8 June 1998 [study days (DSs) 9 to 14), 10 to 15 June 1998 F(DoSgsen1e6rattoi2o1n),ma1l7eJaunnde volumes based on body f1e9m9a8le(DrSats23w)eraenddo1s8eJduwnieth19th9e8 d(oDsSag24e), weights recorded on DS 1 (26 May 1698), all rather than on weekly body weights. As a result of this error, male rats were administered approximately 89% to 90%ofthe targeted dosages taanrdgefteemdadloesraagtessw.erTehiasdmdienviisattieorneddiadppnrotoxaidmvaetresleyly95af%fetcot or interpretation of the study because the precohabitation 99% of the the outcome dosage period was extended for an additional two weeks to ensure that all rats were administered the correct dosages before cohabitation. 3. On 21 August 1998 (postweaning day 1). F1 generation male rats 13158 and 13159 in the 0.4 mg/kg/day dosage group were neither weighed nor dosed. The dosage volume for the rest of the week was abdavseerdseolny tahfefepctostthweeoaunticnogmdeayor2inbtoedryprewteaitgihotn. This of the dsetvuidaytiboencdaiudsneota desuvafayfliu2caipetnoetsttnhwiusemapbnaeirrnagmoefwtepeiorgs,httawnewadenrtiehnegpdrdoeassyaug1meebdvoodtlyoubwmeeeisvgehwrteysrsweiembrialeasrecotdlolodenoctstehadeg.teo volumes based on the day 1 postweaning weight. All deviations Focumented in the raw data. 4 LH = ww 9 / 7 Ri nd G. York/Ph.p., DABT Associate Director,of Research and Study Director Date 3 000 APPENDIX H TEMPERATURE AND RELATIVE HUMIDITY REPORTS 318 GOO48 418-008:PAGE H-1 ARGUS TemperatureLoacnadtiRoenl:atRioveomHum1i7dity Report Protocol Number: 418-008 Range of Dates: 12-May-1998 12:00 to 12-Jun-1998 09:46 STpaercgieetsR:aRnagte: TToottaall NNuummbbeerr ooff HDoauyrss:: Total Number of Data Points: T8e4mFpleora7t9urFe 4312 5 2 | Rela3t0i%vetoHu7m0i%dity 74312.5 742 Mean (2 SD): MMeadxiiamnu:m: Minimum; NNuummbbeerr ooff PPooiinnttss HinigRhan(%g)e: (%): Number of Points Low (%): 696 (11) | 462 (39 6m9s7 641825 664 37s 20 (10)00)| M02 (10900) 0 0) o oo) Report Generated: 04-Sep-1998 at 13:18 COMMENTS: --- -_-- -_ -_ REVIEWED BY: \ 44. L.2 0 DATE: 3/7/51 `Cumulative by Location (v04.01.97) 19 GOO ARGUS 418-008:PAGE H-2 TemperatureLoacnadtiRoenl:atRioveomHu0m3idity Report Protocol Number: 418-008 Range of Dates: 12-Jun-1998 09:46 to 21-Sep-1998 13:40 STpaorcgieetsR:aRnagte: TTToootttaaalll NNNuuummmbbbeeerrr ooofffDDHaoatuyarss:P:oints: Temperature 64Fto 79F 20221107274 Relative Humidity 30% to 70% 22217703274 Maan (50): MMeadxiiamnu:m: Minimum: 00 20| sts ap 1e5t3 20 7763213 Number of Points in Range (%): Number of Points High (%): Numob fPe oinrts Low (%): 2405 0 18 (99.3) (0.0) .n 2404 19 0 (99.2) (0.8) (0.0) Report Generated: 31-Mar-1999 a 08:02 COMMENTS: ---- -_-- -_-- -_-- REVIEWED BY:_e _ clov n/tes DATE: 3-30 aq Cumulative by Location (v04.01.57) 000 ATS ARGUS 418-008:PAGE H-3 Temperature and Relative Humidity Report Location: Room 05 Protocol Number: 418-008 Range of Dates: 27-Jul-1998 12:50 to 28-Dec-1998 08:29 `TSapregceitesR:aRnagte: TToottaall NNumubemroboffDeHaoyursr:s: `Total Number of Data Points: Temperature B40 79F | Relative Humidity 30% to 70% 155 6971.726 369115.526 677 Mean (SD): MMaedxiiamnu:m: Minimum: NNuummbbeerr of of PPooiinnttss HinigRhan(%g)e: (%): Number of Points Low (%): 603 (18) | 563 en 827 69.7 73 58.1 846 43 661 16 (996) | 0.4) 3667 10 (90.7) 03) o 0) o 0) Report Generated: 31-Mar-1999 at 09:22 COMMENTS: -_-- _--_-- -_-- _-- REVIEWED BY: __s=..e . r Yco2 DATE: 2 3-2 3-5 Cumulative by Location (v04.01.97) #3! GOO 418-008:PAGE H-4 ARGUS Temperature and Relative Humidity Report Location: Room 35-37 Protocol Number: 418-008 Range of Dates: 21-Sep-1998 13:40 to 28-Dec-1998 08:29 STpaercgieetsR:ange: TTToootttaaall NNNuuummbbmearroobofffeDDHaaortyuasrs:P:oints: TSemRpeTraItuEre || Relaotivte HuTmoidiy niEeoHs nriyes Mean i 0): 1 won| 25 ao MMMaeidxniiiammnu:om:: oa0s7 E5eH7s NNNumbubermrooobfffPPpeoooiiinnnrtttsss LHiniogRwha(nGgo4er (%): mooefoa(n0n00)||| 24B2T g8Ga3on Report Generated: 31-Mar 1099 at 08:52 comments: -- -_-- -_ revieweo sv: C-af 7]Ann YIN J U oare: 331 Cumulative by Location (v04.01.97) 32 voor 418-008:PAGE H-5 _--_---- ARGUS TemperLaotcautrieonD:evRiaotoimon0s3Report -_-- Protocol Number: 418-008 0 _C7 RangeofDates: TT 12-Jun-1998P 09:460to221. -OSeepp-T1e9e9s8t13:a 40 STpeemcpieersa:tuRarte Target Range: 84F to 79F 7NuD-at1e998 T0i30m0e 17ur1988 0400 Tesmapg.l eas 03VDual-t1e998 TOi0m0e Teem2p4.L 717JJu0-n11999888 0058:0000 17-Jun-1988 0700 s3220LL e27L 22J3uunn11999988 00750000 euu1998 21:00 eeaso7lLL asl 0089u18se88 00Ju1998 22230000 0000 e2Es3l2LL e26L 000801J3u91k198999888 00210000 0300 6e2a5s0L 220 _ 000J0JJuuuiFi119998%9888 000854000000 es6E222r001L0L H=Valus out of raTnegmep-.Hi=gThem_peLra=tuVraeluoeFout of range - Low Report Generated: 31-Mar-1998 at 08:09 These deitons id not avery affect th outcome treatin fh sty. "The following devistion(s) impacted on the outcome of the study asdescribed: Study Ze : ; -- Date: _3/-Mre 99 Deviations by Location (v04.01.97) GOO Pm 418-008:PAGE H-6 = ARGUS Relative Humidity DeviationsReport Location: Room 03 - 0 Protocol Number: 418-008 RRaanndgoeooffDDaatteess::1122-JJuunn--11999880039::4456ttoo2211.--SSeepp--119939881133::4400 Humidity Target Range: Species: Rat 30% to 70% FsChDuaoartievneoets TiaBJimm%cee ot RH. mmaenn Date Time RH. SFairee 3% Jan TraRaodgvmieier moom ve mmonn lan PBIiaaaniseisite pmu1xmm% mmmmeeaonwnn s Hemi ne e nnga. HiValueouofange High =Valueoutofrange Low R.H. = Relative Humidity (%) TTooee locwoivntgodnistiioenssmpvaceaclanthectococmerofveacytatdeosfcsr y Study -- z = : ; ee Deviations by Location (v04.01. .87) Date: _3/ Maz 99 23 COOL 418-008:PAGE H-7 - 00 0 AR- GUS 0 Temperature Deviations Report Location: Room 05 Protocol Number: 418-008 h Pangoe ofDates:27um 1938 12:80 e to 28 Dec 1993s 08:29. RangeofDates: 27-Jul-1998 12:50 to 28-Dec-1998 08:29 Temperature Target Range: Species: Rat 64F to 79F sfforDebaeevtcoeaiatec TiTomnee pEmE ETmIeLTomempien.: Date Time Temp. DBSBiliioooouwwwmse Gmoomkmm n=EEaienn pDDDroioewvrsiimse Eaoa2mmm8 awoneaesny BDDPoooiromeimesm e5wwmm8mo awanioisnnb -= H=oVutoafanougoeA n Lovet ot oframet Repos Cone 1. 168 1 530 Thhsoldlionngsdeviarntt)simvpacttotaanckn uctomaemofovresaoyrats douo af sy, oo L -- Date: 31-Mrp 59 DeviationsbyLocation (v04.01.97) 000 At 418-008:PAGE H-8 mrARGe US ere. Relative Humidity Deviations Report Location: Room 05 Protocol Number: 418-008 re So tO re orm RangeofDates: 27-Jul-1998 12:50to 28-Dec-199808:29 Humidity Target Range: Species: Rat 30% to 70% Eia mE3pDamEtmEEiemlElTABBiimEeoEAaRRRRHe.n mei Em Date Time RH. | _-- | I morte rmOEs es ttn ne toe These deviations did notadverselyaffect the outcome or imynterpretationof the:a! Ee He FE -- Deviations by Location 104.0197) ova 418-008:PAGE H-9 ARGUS RelativLeoHcuamtiidoni:tyRDoeovimat3i5o-n3s7 Report Protocol Number: 418-008 RangeofDates: 21-Sep-19 98 13:" 40 to 28-Deec-c19m98 0e8:29 SHpuemciideist:y RTaatrget Range: 30% to 70% 23-SDeapt-e1998 T10i:m00e 23-Sep-1998 11:00 R2H3.1 2730 2233--SSeepp--11999988 1132::0000 2288201 Date Time RH. H = Value outofrRaHn.ge= -ReHliagthive HLum=idViatlyue(%o)ut of range - Low Report Generated: 31-Mar-1999 at 10:12 Thess devitons didnot adversely aft he outcome or interpretationothe study. The following deviation(s) impacted onthe outcome of the study as described: Study Director. i , s = Deviations by Location (v04.01.67) Date: _5/-uap 59 33% 000716 APPENDIX| STATEMENT OF THE STUDYDIRECTOR 000s er en -- 418-008:PAGE I-1 PROTOCOL 418-008: COMBINED ORAL (GAVAGE) FERTILITY, DEVELOPMENTAL ANDPERINATAL/POSTNATAL REPRODUCTION TOXICITY STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: 6295.9 STATEMENT OF THE STUDY.DIRECTOR perforTmhainscefinoafl trheeposrttudayc. cuNraotedleyviraetfiloecntss ftrhoem rtahwe dataobtained during U.S. Food andDrug the Administration (FDA) Good Laboratory Practice Regulations; Final Rule', the Japanese Standard Ministry of Health for Safety Studies oanndDrWueglsf"araend(MtHheW)EuGroopoedaLnabEocroantoomriycPrCaocmtmicuenity (EEC) Council decision on 28 July 1989 on the acceptance by the European pErcioncniopmliescCoofmgmouondiltayboroaftaonryOprEaCctDicde*ecoicsciuornr/erdetchoamtmaefnfdeacttiedonthoenq`ucaolmiptylioarnce with integrity of the study. Carlen , Raymond G. York, Bh.D., DABT Date Associate Director of Research and Study Director 1 Argus Research Laboratories, Inc. a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58, b. Japanese Ministry of Health and Welfare (1997). Good.Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. C. European Economic`Community (1989). Council decision on 28July 1989 on the acceptance by the European Economic Communityof 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. 3 COOEES APPENDIX J `QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT 40 GOUA49 r~PRIMEDICA 418-008:PAGE J-1 SalTeapsphodm A e Argus Research Laboratories, Ic. `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 418-008: Combined Oral (Gavage)Fertility, Developmental and Perinatal/Postnatal ReproductionToxicity Study of PFOS in RSpaotnssor's Study Number: 6295.9 and The draft protocol for this study was audited for Drug Administration (FDA) Good Laboratory Pra adherence to U. S. Food Ministry of Health andWelfare (MHW); ctice Regulations, Japanese GoodLaboratory Practice Standardfor Safety Studies decision on 28 on Drugs, July 1989 and European on the accep Economic Community (1989) council Community of an OECDdecision/recotmamnecnedabtyitohneE`uonrocopmepalniEacncoenowimtihpcrinci of goodlaboratory practice on 13 MAY 98. ples and Critical ra phases of this study were inspected 24 times; study information w data phases/data). were audited six times (see tables 1 and 2 for dates and auditedThfoer adrcacfutrfaicnya,l rfeoproardthaenrdentchee rtoawprdoattoacoflorrethqiusirsetmuednytws,eraend`cfoomrpaadrheedraenndce RtoegUu.lSa.tiFoonosd, JaanpdaDnreusge AMdimniinsitsrytroaftiHoenal(tFhDaAn)dGWoeoldfaLraebo(rMaLtWo)ry: PGroacotdiLcaeboratory Practice Standardfor Safety Studies on Drugs, and Community (1989) council decision on 28 July 19 European Economic European Economic Community of an 89 on the acceptance by the OECDdecision/recommendationon acnomdp2l9iaAnPceRw9i9t,h aprnidncfioprlreesviosfigonoso.drleaqbuoersatteodrybyprtahcetiSce between 05 DEC 98 7 JUN 99, 9 JUN 99, and for finalization on 10 JUN ponsor 99. 18 MAY 99, Coors 418-008:PAGE J-2 AdminisTthriastisotnud(yFDwAa)sGcooondduLcatbeodraatcocroyrdPirnagcttioceU.RSe.guFloaotidonasn,d JDarpuagneseMinistry of Health and Welfare Studies on Drugs, and (MHW); Good Laboratory Practice Standard forSafety European Economic Community (1989) council decision `oOnE2C8DJduelyci1s9i8o9n/ornectohmemaecncdeapttiaonncoenbcyotmhpeliEaunrcoepewiatnh Economic principles Communityof of good an laboratory practice. Fang rghitdin fotos san /. ) Nancy J."Gongliewski Date Quality Assurance Manager Heather L. Rabuttino, M.S. Quality Assurance Supervisor and Principal Auditor Date IL ooo=2 TABLE 1 CRITICAL PHASES INSPECTED 418-008:PAGE J-3 TAesdtAmrtiiclne istrat-Giavoagne Dates of inspection: 28 MAY 98, 02 JUN 98, 03 SEP 98 Dates results reported to the Study Director and Management 15 JUN 98, 15 JUN 98, 18 SEP 98 Estrous Cycle Evaluation Date of inspection: 11 JUN 98 Date results reported to the Study Director and Management: 11 JUN 98 Cohabitation Dates of inspection: 08 JUL 88, 04 NOV 98 Dates results reported to the Study Director and Management: 14 JUL 98, 12 NOV 98 Scheduled Sacrifice - Day 10 Date of inspection: 17 JUL 98 Date results reported to the Study Director and Management: 03 AUG 98 Test Article Preparation Dates of inspection: 29 JUL 98, 16 SEP 98 Da1t4eAsUreGsu9l8t,s 2r4epSorEtPed9t8o the Study Director and Management: Natural Delivery Dates of inspection: 31 JUL 88, 25 NOV 98 Dates results 14 AUG 98, reported to 02 DEC 98 the Study Director and Management: Blood Collection 203 0002 Dates of inspection: 31 JUL 98, 18 AUG 98 418-008:PAGE J4 Dates results 14 AUG 98, reported 21 AUG to 98 the Study Director and Management: Physical Eve and Reflex Opening, Audi Development le, -ASirurfRaigchteingR,ighPtuinpgi,l Pinna Unfolding, Constriction, Sexual Maturation Da1t1eSsEoPf i9n8spection: 31 JUL 98, 13 AUG 98, 18 AUG 98, 28 AUG 98, Da1t4eAsUreGsu9l8t,s 0r3epSorEtPed8t8o, t0h8e SStEuPdy98D,ir1e1ctSoEr Pan9d8,Ma1n2aSgEePme9n8t: MaleNecropsy Date of inspection: 31 JUL 98 Date results reported to the Study Director and Management: 14 AUG 98 Necrops-y Dam and Litter Sacrifice Dates of inspection: 18 AUG 98, 15 DEC 98 D2at1eAsUrGesu9l8t,s 1re8pDorEtCed9t8o the Study Director and Management: Day 21 Weaning Date of inspection: 18 AUG 98 Date results reported to the Study Director and Management: 08 SEP 98 Behavioral Testing - Passive Avoidance, Watermaze, Dates of inspection: 25 AUG 98, 08 OCT 98 Da1t2eSsErPesu9l8t,s 1re2p0orCtTed9t8o the Study Director and Management: Ww 00033 TABLE 2 418-008:PAGE J-5 RAW DATA AUDIT(S) on 2T9hSeEfPoll9o8w,in0g1 sOtCudTy 9i8n,fo0r4maOtiCoTn a9n8dTrOaw05daOtaCTwe9r8e: audited Protocol. Protocol amendments. List of personnel and `computer operator codes. Error codes and codes for clinical sigh observations. IAnn-ifmfealtrraencseaicptt,iornanrdeocomridz.ation, physical examination and acclimation. Feed consumption. ONercgraonpswye.ights. Tissue packing lists. DEdeivtiarteiqounessts. Data review page/pages. KBleoyofdorcotlelsetcitnigonfadcialtitayacnodmppuatcekrinbgaclikstusp. record abbreviations. on The results 06 OCT 98. of this audit were reported to the Study Director andManagement CovER 418-008:PAGE J-6 on 0T6hOeCfoTll9o8w,in1g2sOtuCdTy information and 98, 02 NOV 98 TrOaw0d6aNtaOwVer9e:audited AInn-ilmfealtrraencseaipctt,iornanrdecoomridz.ation, physical examination and acclimation. EFseterduscocyncsluempetvailouna.tion. Cohabitation. MCaateesranraelang-rsoescstioobnsienrgv.ations LNiatttuerraolbsdeerlviavetriyonosb.servations. TPaubpleboodfyrawenidgohmtsunaitnsd. status. Necropsy. GTiesnseurealpaccokmimnegnltisst.s, SRtefuldeyxmaanidntpehnysainccael rdeecvoerldos.pment. TFeeemdp,erwaattuerreaannddberedldaitnivge ahnuamliydsietsy.reports. DEdaittarreeqvueisetwsp.age. PBluopodstcoomllaecchticoonndtaetnatsa.nd packing lists. on 0o8meNOreVsu9l8t.s of this audit were reported to the Study Director and Management GOO Xen 418-008:PAGE J-7 on The following 08 NOV 98 to study information 09 NOV 98: and raw data were audited Vehicle receipt, preparation and use. Test Test article article receipt, packing preparation lists. and use. on The results 12 NOV 98. of this audit were reported to the Study Director and Management on 2"6ThJeAfNol9lo9witnog2s8tJudAyNi9n9f:ormation and raw data were audited In-ife transaction record. PFaesesdivceonasvuomipdtainocne.. Watermaze. NGeecnreoaplsoyg.y chart. Organ weights. STiesxsuuael pMaactukriantgiolni.sts. Edit request. on The results 28 JAN 99. of this audit were reported to the Study Director and Management 000226 418-008:PAGE J-8 The following study information and raw data were audited on 28 JAN 99 to 01 FEB 99: In-life transaction record. Feed consumption. Cohabitation. Natural delivery observations. Litter observations. Pup body weights and status. Passive avoidance. `Watermaze. Table of random units. Necropsy. Tissue packing lists. General comments. Sexual maturation. `Study maintenance records. Temperature and relative humidity Feed, water and bedding analyses. Edit requests. Dosage volumes. Data review pages. reports. The results of this audit were reported to the Study Director and Management on 01 FEB 99. The following study information and raw data were audited on 25 JAN 99: Vehicle receipt, preparation and use. Test article receipt, preparation and use. Test article packing lists. The results of this audit were reported to the Study Director and Management on 28 JAN 99. Jub "OOTY