Document VVy5rYzqN99p693g357D4RbN

. AR 286-149] pp Duron 13267 PROTOCOL Ammonium Perf`lPuoostr-oWocetaannionagteR:atAsgeFoElflfoewcitnognOtrhaelPGlaavsamgaeConcentration in Project Number: DuPont-13267 Work Request Number: 14762" Service Code Number: 1389 ] gsm L332 z 33 > oo Haskell Animal Wel Commits No. BTTISSP Nt Worker Sdeicode1368wetsfore pingpnof0 dy. --e---- Tage Tore8 e-- r ---- ---- -- A`mComnocennitruamtioPneifniPuoosrotoWcetaannionsgtR:aAtsgeFoEllofwionfgntOherealPclGaastvmage `TOAFCBONL TENETS DuPont13267 Page TABLE OF CONTENTS... pm------ INTRODUCTION essere: 3 SPOANNDSTESOTIRNG FACILITY css3 REGULATORY COMPLIANCE crores3 MATERIALS AND METHODS .crccnsssnonssssessssssssssossnssies3 J -------- ---- B ToS ----, 2. ENVONMERtAl CORGHHONS .rvrorrorsrsrsnssnsssndh 4. Animal HealthandEnvironmental MODHOFingPrOGIAI ovcvvvvvrnenndh TY --. 2. DosePreparation,AnalyAsdiRBsES, vernon 3. SacanrdBiIOOfASiAMPcHDeE vrs 5. PFOAARBIYSIS corns SAFETY AND HOUSEKEEPING crores RECORDS AND SAMPLE STORAGE ..rcrrorsrrsssssisninssssnsss PROPOSED STUDY DATES crams | BETERERCER cms A------ -_ En A`mmConoinumoPeeiclnnuPctorsotaocWteiaannooisnnigs:RaAtgseFEoflolfntoOhereswPcliGaasnvtmaagge Duron 13267 INTRODUCTION Ainmdumsotrniailusmurpfearctfalnuto,roEolcitmainnoaattieo(nPoFfOPAF)OAis isnyrnathteiticsepxe-rfdleupoernidneanttedaoncdthanoorimcaocniedruesgeuldataesd.an PFOA elimination is much testosterone in both female faster in female rats than in and castrated male rats," male rats," down-regulated by and up-regulated by estradiol in the male rSattusdy isHtoowdeevteerr,mitnheeisfe sitmumdaitesurweermealdeonaendonfesmeaxluealaltysmaeltiumrienartates.PFTOheAofbrjoecmtbilveooofdtahteacruartreent dwiiflflebreendtotsheadn wtihatthoPbFseOrAveadnidn tmhaetpulraesrmatas.conMcaelnetraantidofnesmoaflPepFoOstA-waeta2n4inhgourrastspoasttd-idfofserienngtwaiglelsbe cPoFmOpaAreedli.miTnhaitsiotnimine mpaoitnutrewarastschcoasnebnebeeacsaiulysedi2s4tihnoguurisshaefdtvridaotshiengp,lathsemasecxondciefnfterraetnicoeninof PmaFtuOrAe.ratsT,haisndstiusdnyotisdneostigdneesdigfnoerdustoe fiunldlyetcehramraicntienrgizdeeftihneitpihvaeremlaicmoiknianteitoincrsatoefcPoFnsOtAantisn. SPONSOR AND TESTING FACILITY "WTihlismisntgutdoyni,s Dceol-aswpaornes.oreTdhbeysp3oMn,soSrt'.sPaauplp,roMvNal,waansdeEf.fLe.cdtuivPeotnhteddeatNeetmhoeusrpsonasnodr Cauotmhpoarinzye,d the work on the contract. TEhLedsutuPdoynwtildlebNeemcoonudrusctaenddaCtoHmapskaenlyl,LNaebworaartko,ryDefloarwHaerael,tUhSaAnd. Environmental Sciences, REGULATORY COMPLIANCE `LTahbiosrsattuodryywPirlalcbtiecceoSntdaunctdeadrdisn.compliance with U.S. EPA TSCA (40 CFR part 792) Good : MATERIALS AND METHODS A. Test Substance "HP-F2O49A2w1ilulpboenorbetcaeiipnte.d fArvaoimla3bMle(iStn.foPraumla,tiMoNn )onatnhdeapsusriitgyn,edcoHmapsokseiltlioLna,bocroanttoarmyinNaunmtsb,er sdyoncounmyemnst,edhaiznartdhse,staunddyhraezcaorrddosuasnmda/toerrrieaplorctl.assification(s) will be provided by the vendor and CASRegistry Number: 335-67-1 Molecular Weight: 4141 Molecular Formula: ~~CuFisOsH ------------------------------eEeReeeeeeeeeeesiee. `AComnwceonstriaotimonPienriPvoosrtooWceasnnoisntge:RaAsgeFoBlelocwtiongnOtrhalPGlasviaage DuPont13267 B. Test System MInacl.eRaanldeifgeh,maNloerCthrlC:arCoDlin(aS.D)TIhGeSBSRprraagtsuewi-lDlabweleoybtraatinweadsfcrhoomsCehnafrolresthRisivsetrudLyabboercaatuosreieosf,the eTxotwenisnicviedeenxcpeeorfisenpcoentwaintehotuhsisdsitsreaaisneas.ndFiutrstshueirtmaboirlei,tytwhietShprreasgpuecet-Dtaowlloengyervaittyh,assebnseietnivuistye,dand previously for toxicokinetic testing ofPFOA and other fluorinated test materials. Avatritahteiotnimsehoofudlodsniontg,exrcaetsedsho2ul0d%obfetahpeprmoxeiamnatweeliygh4,t 5b,y6d,o7s,eagnrdo8upwaenedksseox.fagIenfaonrdmatthieowneoinght {ahsesicgangeedltaobeelascwhirlalt.inTchleudientdhieviHdousaklelidlenatniifmiaclatniounmnbuemrbaenrdwainllibndeipvlidaucaeld iodnentthiefiacialtoiofncnauchmbreart. C. Animal Husbandry 1. Avimal Housing UapppornoparriraitvealcaagteHsasakceclolrdLianbgortaotSotrayn,draartds wOiplelrbaetirnegmPorvoecdedfurroems suahliepspsinsgpeccairftioendsianntdhehosutsueddy in roetchoerrdwsi.seAbnyimtahlessiwtiellvebteermianairnitaani,nheadvuendaterleqaustaroannetirneecoforrdeadt lweeaisgthtthrgeaeind,ayasndunnloesasbnaoprpmraolivteides detected. After the quarantine period, ats will be selected for study. TahpprrooupgrhiaotuetctahgeedsoascicnogrpdeirnigodtowtihtehSteOsPt.substance, th rats will be housed individually in 2. Environmental Conditions Arenliamtiavle rhouommi(dsi)tywoifll3b0e-m7a0i%nt(atianrgeedteatdatote4m0p-e6r0a%t)u.reAonfim1a8-l2r6ooCm(stawrigleltbedetaort2i2fi-c2i4allCy)iallnudmainated (fluorescent light) on an approximate 12 hour light/dark cycle. Unless judgedby the study dreilraetcitvoerhourmtihdeiltaybaornadtotreympveertaertiunraerriaanngteoshianvethseihgnoiufsiicnangtrlyooamffsecwtieldltbheerreecsourltdseodfbtuhtewisltludnyo,ttbhee included in the final report. 3. Feed and Water All animals will be provided tap water ad libitum and fed PMI Nutrition International, LLC C1e2r1t1i5fiheoduRrosdbeenftorLeadboDsiientgw5i0t0h2teasdt lsiubbisttuamn.ce.AnFiomoadlswwiilllbebereftausrtneeddoavperpnriogxhitmaftoerlaypptrwooxhiomuartsely post-dose. 4. Animal Health and Environmental Monitoring Program AFsollspoewciinfgipedroicnetdhuereHsasakreelpleLrafboorrmaetdorpyerainoidmicaalllhyeatlotheansnudreentvhiartocnomnetnatmailnamnotnilteoverlisngarperboeglraomw, the those that would be expected to impact the scientific integityofthestudy: -_ ER `AComncmenotuntioPneirnfPsocsrtoWocetanpoisntg:RAagseFEolfloewionng OtrhaPGlaivsmaage Dupont13267 + aWnadteorthsearmcpolnetsamairneaanntsa.lyzed for total bacterial counts, and the presenceofcoliforms, lead, + Feed samplesareanalyzedfortotal bacterial, spore, andfungalcounts. + SSaanmiptlateisonfbryomthferecsahgleywwaasshheresd.cagesand cage racksareanalyzed to ensure adequate rCeerqtuiifrieemdeanntismaanldfneoetd tisouesxecde,egdusaartaendtemedaxbiymtuhemmcaonnucfeancttruatrieorntsooffmkeeetyscpoencitfaimeidnannuttrsi,tiionncalluding PSpreecsiefniceedohfetahveysemectoanltsa,maifnlaantotxsibn,eclholwortihneamteadxhiymdruomcacrobnocnesn,traantdioonrsgaantoepdhobsypthhaetemsa.nufTahceturer Would not be expected to impact the integrityofthe study. "lTahbeoraantiomraylanheiamlatlhvaentdereinnavriiraonn.meDnattaalamroenmiationrtianignperdosgerpaamraitselaydmfirnoimstsetrueddybryectohredastatnenddminagy be included in the final report atthe discretionofthe study director. D. Main Study 1. Animals "The experiments will be composedoffive animal groups: T Group ET TawT Male Fem-- ale 1' m 54 3xs 5 2 1100 1100 10 10 Nvv 57 a5 1100 1100 ------ ee ---- ------ r----ee er =Anoivimnaglwsiwillblebdeoc1udmaeyntfeodimntthheetsarugdetyraegceordtahnedtliomrefoifldorseinpgr..Thesugle t 2. Dose Preparation, Analysis, and Rates "isThtehetersotustuebmsotasntcceowimlmlobnelyadumsienidsftoerretodxbicyistyinsgtluedioersalofgaPvaFgOeA..TPhiFsOrAouwteilwabse cmhioxseednwbietchawuasteeirt etoquaicvhaileevnettthoe1t0armgegtPdFosOeA/cokngcbenotdryatiwoenisg.htFoarndaltlheexdpeorsiemveontlsu,mea wsiinlgllbeedaopsperloexviemlawtielllyb4emuLse/dkg body weight. ``SAtcutduyarlecdoorsdesl,eavneldpmraeysbeenmtoedidfiinethde fatintahereSptourdt.y DDiorsecetmoar'ysbdeispcrreteiopn,awoirlnletbhdeeddoacyumofeunsteeodripnrtihoer fo the dayofuse and stored under appropriate conditions. - EE `ACmomncoenntiruatmioPneirnfPloosrtoWoecatnmoisntg;RaAtgseFEofflecolntOotrhaewPGileavsnamgage Dupont13267 'HPLC-MS methods will be used to confirm PFOA concentration in the dose solutions. 3. Sacrifice and Blood Sampling Rats will be sacrificed by CO; asphyxiation and exsanguinated by cardiac puncture. 4. Experimental Procedure TRawtesntwyi-llfobuerohbotuarisneadftferroPmFtOheAvaednmdionirsatrnadtiaodnm,inainsitmearlesdwPilFlObAe sataacridfoisceedlaevnedlwdheosclreibbeldooadbove. staubmepsleasndwimlalibnetacionleledcotnedwbeytciacre.diPaclapsunmcatuwriel.l beWhsoelpaerbatleododbysacmepntlreisfuwgialtliboen palnadcferdoiznetnoaEtDTA <-10C until analysis. SKtiodrneedyasta<n-d60livCerfsorwiplolsbsiebcloelfluetcutreedabioscahcerimfiiccaelaanndalwyislils.be flash frozen in liquid nitcogen and Rfaotrstthheaptraerseefnocueonfdddieseaadsoer.arDeespaecrnidfeincteduipnonextthreemfiinsdwiinlgsl,befugtihveerndaiaggrnoosssteixcapmrioncaetdiuornetsomcahyebcek Veemtpelroiyareidaant.tThehedirsecsruelttisonwoifllthnoetsbteudiyncdliurdecetdori,n tahpeatfihnoallogriespto,rot untlheeslsacboonrastiodreyreadnismiganlificant to the evaluationofthe study. 5. PFOA Analysis tHhPeLmCe-thMoSdmseeltehcotdedwviillllbbeeiunsceldutdoeddeitnertmhienfeinpallarsempaorctoncentration ofPFOA. A description of E. Statistical Analysis GasrsoeuspseddabtaywtihlelSbteudreenptr'essetn-tteesdt.aOttMheerastna+tiSsDt.icaWlheevraleuaatpipornopsrmiaatye,sbteatpiesrtfiocarlmseidg,niiffincecaenscseawriyl.l be SAFETY AND HOUSEKEEPING MAlSl DchSemaincdaldsisupsoesdeddoufriancgcothridsisntgudtoy twhiellSbteinhea-nHdaslkeedllacWcaosrtdienDgitsopotshealprGoucieddeulriensessapencdiftiheedairneathe Safety, HeanadEnl virot nmenhtal (SHE) manual. RECORDS AND SAMPLE STORAGE ``NSepweacrikm,enDselGafwaaprplei,coabrlaet),IrroanwMdoautnat,aainndRtehceofridnaslMraenpoargtewmielnltb,eWrieltamiinnegdtaotn,HaDseklealwlarLea.boratory, J--T------- = =--m--MmM `ACmomnocneinutmioPneirfPlocsrtooWcetaannicnagt:RAagssFEoflfleocwtionngtOherPslGaasvmaage PROPOSED STUDY DATES DuPont13267 Dateof Approval: Proposed Experimental Start: Date Study Director signed Protocol June 30,2003 Proposed Experimental Termination: ~ August 29, 2003 Proposed Study Completion: `December 18, 2003 REFERENCES 1. tOohxmiocrokiinKe.t,iKcsudboe,twH.e,eKnaptearyfalumoar,ocKa.r,baonxdyiKcaawcaisdhsiwmiat,h Yd.if(f2e0r0e3n).carCboomnpacrhaiisnonloenfgtthh.e Toxicology 184, 135-140. 2. oVfapnedrefnluHoeruovoeclt,anJo.iP.c,aDcaivdiisn, mJa.Wl.e,rSatosm:mienrhisb,itRo.r,yanefdfePctetoefrstoens,toRstEe.ron(e1.99J.2)BiRoecnhaelme.xcTroexticiooln. 7,31:36. 3. `KCuodmop,arN,i,soSnuozfuktih,eE.e,liKmaitnaatkiuorne,beMt,,weOebnmpoerrif,luKo.r,inNaotsehditfoa,ttRy.a,ciadnsdwKiatwhadsihfifemrae,nt.ca(r2b0o0n1c)hain length in rats. Chem. Biol. Interact. 134, 203-216. 4. tYhieinuerni,naMr,y eHxacnrheitjiaornvoif,pHe.rfJlauaokroonoacthaon,oJi,c aacniddPienurtah,e mP.al(e19r8at9.)PShtairmmualcaotli.onTboyxioceoslt.ra6d5i,ol27o4f27. 5. UDnupPuobnltisHhaesdkerelplorLta,bDourPaotnoirty-7(427030.3). Perfluorooctanoic Acid: Toxicokinetics in the Rat. 6. U.S. EPAGoodLaboratory Practice Standard, TSCA (40CFR 792). (1989) 7. EDxupPoosnurteH,asUkneplulbLlaibsohreadtorreypo.r(t1.997). Hazard Characterizationfor Human Health C8 8. dViasntrdiebuntiHoenu,vmeelt,aJb.oPl,isKums,liaknids,elBi.mLi,naVtainonoRaffpeelrgfhleumo,roMo.cJta,naoincdaPceitderisnonm,alReEa.n(d1f9e91m)a.leTriastss.ue J. Biochem. Toxicol. 6, 83-92. - ER ------------------ AmmoniumPefluorooctanost: Age Bit onthe Plasma Concentration i PostWoening RatsFollowing OnlGevege SIGNATURES DuPont13267 f o`Wrriltatseuneabnyd Appro Study v ed . [Phstiot E ingHone , PhD. Research Toxicologist feDuses Approved by: =A TPriT ncipalRGesaerayrcWh.TJoecposloong,iPshtDa.nd Managsc 27-Tim 2002 Dae CC: IRCWHaRmiilcla, ,HAHSAK.SKHLHI PMGLMSKHDeonignniedadneyi,ry,B,AHHSAAKS,SK.,FI1HL LSAA GMaaninonng,,HHAASSKK,.HHlI LLSGMaFBzouurrcmdhdfaHirAe,lSdBK,A.BSAHKSL.K.HIH1 e-- e -------------- = -------------- i ------------------