Document YDww4kNZp4qko0L08ObN4NzNy

PPFFOOAA::LLaactcattaitoinonaannddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccStdudyyiin RnatsRats____DDuuPpoonnt.1313330099 AR22b- 155 | ~ pos le lbeadoal al, ir " TRADPSECRET Gow Budenho LE Gl Gren Study Title PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats Laboratory Project ID: DuPont-13309 T6335.) AUTHOR: Eve Mylchreest, Ph.D. STUDY COMPLETED ON: December 19, 2003 PERFORMING LABORATORY: EHIa.skdelulPLoanbtodreatNoerymofuorrsHeaanldthCaonmdpEannvyironmental~S~ciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 SPONSOR: 3M Company 3M Center St. Paul, MN 55144-1000 and EW.iLlmdiungPtoonnt,dDeeNleawmaoruers1a9n8d98Company WORK REQUEST NUMBER: 14787 SERVICE CODE NUMBER: 1569 TTT TT T hmia PPFFOOAA::LLacatcuttaitoinoansnddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSutuddyy iinnRRaass DDuuPPoonntt.1133330099. GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT `This study was conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards, which are consistent with the OECD Principles of Good Laboratory Practice (as revised in 1997) published in [ENV/MC/CHEM(98)17 and MAFF Japan Good Laboratory Practice Standards (59 NohSan No. 3850). Appli/ cSpaonnsotr: ELL du Pont de Nemours and Company `Wilmington, Delaware 19898 USA. and 3M Company 3M Center St. Paul, MN55144-1000 \ Study Director: fehreest, PhD. flor Research Toxicologist (7- Dee- 03 Dai Applicant / Sponsor: "ApplicantSponsor Representative Date -Y PPFFOOAA::LaLaccttaartiioonnaallsannddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSStuuddyyiian RRaattss__ QUALITY ASSURANCE STATEMENT DDuubPomnt1313330099. Haskell Sample Number(s): 24921 Dates of Inspections: Protocol: July 11,2003 Conduct: Records, Reports: July 11,28,31, 2003; August October 2,3,6-8,13-15,30-31 15, 2003; September 5,30 2003 2003; November 3-7, 2003 Dates Findings Reported to: Study Director: July 11,2931, 2003; August 18, 2003; September 10,30, 2003; October 10,16,17, 2003; November 67,10, 2003 . Management: JOucltyob2e9,r311,0,21060,137;,A2u0g0u3s;tN1o8v,e2m0b0e3;r S6e,p7,t1e0m,b1e2r,21000.335,2003; Reportedby: Tle Phe Ln 1B Kimberly B. Brebner Staff Quality Assurance Auditor 19-OEc 2062 Date 7 -- -- i FPEFOOA:: LLaaccttaaitoinoan)aannddPlPalceanctaelTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSttuuddyyiinnRRasts__DDuupbonmt113333059 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluationofdata obtained from this study. Anaya Evaustionsby: (AptTanerP.lVipshaankts,aBrS.da Senior StaffChernist i= dTiTe -z003 Biochemical Evauations by: SurShaAwn AFGanpnonn, BeS n Senior Staff Toxcologit $= Ye = 2003 Dac Arprove by: fos Friky ReseSaercohnM,anLoavgeelresasn,dPDhiDr.ector 1 -DEC ~2oe3 Date Issued by Study Director: of Resce/Hraesrch TosiPcho.lDo.gist T 19-No ee- 2w 03 PFOA: Lactatonal and Placental Transport Pharmacokinetic Study inRats DuPont 13309 TABLE OF CONTENTS Page GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT... QUALITY ASSURANCE STATEMENT... snd nd CERTIFICATION... ------ rh LIST OF TABLES . ---------- --. LIST OF FIGURES. ---------- mend STUDYINFORMATION wns mmm STUDY PERSONNEL wc mmmm-- SUMMARY. mmmmm-------- INTRODUCTION... wmm-------- OBJECTIVE... smm---------- . SPONSOR ANDTEST FACILITY ma---------- - STUDY DESIGN.............. ---- red) A. Experimental Design. ......vromrorrr . rrr 13 B.. Selectionof Dose Levels... erp ---------- 14 MATEANRD MIETHAODSL ....S..... n---- ell ANALYTICAL........ ------ A VOR vsmmnmmmen-- nsmmnebtl nmm-------- B. Test SUBSIANCE wc. n------ nit INLIFE.cc. A TeSU SPECIES rrr ---- -------- onl] comment] B. AnimalHusbandry..... -------- C.. QUAaNAdPRNRES!IPETNIOM resem A 19 D. Assignment to Groups ~ Randomization....... -- E. Inlife OBSerVations........ov. F. Terminal Sample Collection and SACHfICe mmismmem----r mtd BIOAC.HTEiMssIuCeAaLndTPOIXaIsmCaOALNOGIGYYSISS.A.M..P.L..E..A.NoAoLoY.SES.......c. wm----m--s ----m ----]r STATISTICAL ANALYSES... nnn 23 RESULTS AND CONCLUSIONS.......... ------ wml ANA.ATeLst SYubTsDtAaInTcAeCc StAabilLity ANALt YSES... io I n .25| B. C. ATNeIstYUSCuHbS]aCnOcNeCIFUoSrImOuNlSatr ion ARISES .s ......veurems remssmrnosresiore eros 25 27 PFOA: Lactational and Placental Transport Pharmacokinetic Sudy in Rats DuPont 13309 REPRODUCTIVE TOXICITY EVALUATIONS we. wemmsnnnddl IN-LIFE TOXICOLOGY cr m--------l A. MaternalObservations. ---------- ---- B.. REPIDUOR IrcsUssIsssVsssEsnssssmsrommmereneereeeeeeeeeeone 3 BIOCHEMICAL TOXICOLOGY ---------------- nl A. Maternal, Fetal, and PUp PLAST... 30 Be. Millipunsmmmmmmnmmisssssmmmmmesseemmm3e C. AmnioticFluid........... ---------------- orn 30 D. Ee Placenta EIDIYO AE FOIUS m------ snm---- crosses 30 CONCLUBLIIS mimi -------- sommendSl RECORDS AND SAMPLE STORAGE... WO. REFERENCES... s-------------------- TABLES mms r-- mim FIGURES wenn ss-------------------- APPENDICES.......... ----------------------------) LIST OF TABLES N,No Page TABLE | SUMMARYOFDOSINGTEST FORMULATIONANALYSES... 0 `TAPBOLS2ETPCAORNTCUEMN.TRATION OF PFOAr--IN--MATERNAL PLASMA DURINrGerGeE----STATION AND -- `TABLE 3 CONCEOFPNFOATINFRETAALANTDPI UP ON PLASMA.. on 3T TABLE 4 CONCENTOFRPFAOATINION MILK.. sm TABLE CONCENTRATION OF PFOA IN AMNIOTIC FLUID A 3 `TABLE 6 CONCENOTFPRFOAAINTION PLACENTA... A `TABLE 7 CONCENTRATION OF PFOA IN EMBRYOFETUS... --a PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont 13309 LIST OF FIGURES Page FIGURE | MEAN BODY WEIGHTS OF FEMALE RATS DURING GESTATION... " FIGURE 2 MEAN BODY WEIGHTS OF FEMALE RATS DURING LACTATION nd FIGUDRUERI3NCGOGNECSETNATTRIAOTNIOANNDOFPPOFOSATINPMAATRETRNUAML.PL.A.SMA nn FIGUR4E CONCENTRATION OF PFOA IN PUP PLASMA CR a FIGURE $ CONCENTRATION OF PFOA IN FETAL PLASMA ON GESTATION DAY 2148 FIGURE 6 CONCENTRATION OF PFOA IN MILK. mmm----_y FIGUR7E CONCENTRATION OF PFOA IN AMNIOTIC FLUID... 50 FIGURE CONCENTRATION OF PFOA IN PLACENTA ...... WN FIGURE 9CONCENTRATION OF PFOA IN EMBRYOFETUS. wns LIST OF APPENDICES . Page APPENDIX A ANALYTICAL DATA cc wg APPEDNUDRIIXNGBGIENSDTIAVTIIDOUNALACNLDINLIACCATLATOIBOSNE.RV..A..T.I..ONS AND MORTALITY e DATAmem~ --r APPENDIX C INDIVIDUAL BODY WEIGHTS DURING GESTATION AND LACTATION -- APPENDIX D INDIVIDUAL BODY WEIGHT CHANGES DURING GESTATION AND LACTATION.......102 APPENDIX E INDIVIDUAL REPRODUCTIVE DATA wennn-- 2 APPENDIX F NUMBER OF PUPS DURING LACTATION... 18 APPENDIX G INDIVIDUAL LITTER CLINICAL OBSERVATIONS ES APPENDIX H MEAN PUP WEIGHTS PER LITTER. omem-- 123 APPENDIX PLASMA AND TISSUE ANALYSIS... rn 126 PFOA: Lactation nd Placenal Transport Pharmacokinetic Study in Rats DuPont13309 STUDY INFORMATION 9th Collective Nomenelaure: Octanoic acid, pentadecafluoro-, arumonium salt Synonyms/Codes: ++ AFCm-m1o4n3iFuLmUpOerRfAluDorBoroactnadnoFaltueorochemical Surfactant (3M . cCosmpany, Specialty Materials) +PPFerOflAuorooctanoate, ammonium salt + HLo2t032321(3M Specialty Materials) (Lot No.) HaskellNumber: 24921 CAS Registry Number: 382526-1 Purity S9u5w.a2ig%ht-c9h7ai.n9:9%77.6% Branched: 192.%6%isionptreompayll monomethyl (non-alpha) 00.21%% geermt-bdiumyelthyl 0.1% alpha monomethy! Known Impurities: Physical Characteristics: Stability: CC,L((CC5IFF;,CCOO;;" NNHHLL)),, 00.0031%% ha CCra((CCiiFFuyCCOO;; NNHHLL)),, 004537%% CMo3n(oChiyFdurCoO;APNFHOL,).0.00.91%6% MUonndoeufnisnaetdu(rpaotsesidblAyP)FsOu,bst0i.t7u2te%d perfluoroeyclo species, 02% cyclopentyl, and 0.19% cyclobex! White solid Tthheestteusdty;sumbosteavnicdeenacpepeoafreindsttoabbielisttyawbales uonbdseerrvteh.e conditions of `Spenser: EWiLlmdiungPtoonnt,dDeeNleawmaoruers19a3n9d8Company UanSd A 33MM CCeonmtpearny St.Paul, MN 55144-1000 `Study Initised/Completed: July 8, 2003 (see report cover page) In-Life Initised/Completed: July 11,2003/ August 22, 2003 -- 5 -- PFOA: Lactational and Placental Transport Pharmacokinetic Sudy in Rats STUDY PERSONNEL Study Director: Eve Mylchreest, Ph.D. Management: Scott E. Loveless, Ph.D. Supervisor: Deborah L. Tyler Primary Technician: Joseph F. Aschiero Analytical Chemist: Janet C. Maslanka, B.S. Management: S. Mark Kennedy, Ph.D. Biochemical Toxicologist: Shawn A. Gannon, B.S. Management: Gary W. Jepson, Ph.D. Toxicology Report Preparation: Mary K. LaRoe LisaG. Burchfield, A.A. Management: Nancy S. Selzer, M.S. Laboratory Veterinarian: Thomas W. Mayer, D.V.M., A.C.LLAM. Management: JaniceL. Connell, M.S., B.A. DuPont13309 . \,A 5 -- -- PPFFOOAA:;LLacatcattaiioonnaall aannddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSwdtyiuinndRRaatytss __DDuuPPoonntt1133330099 SUMMARY `coTnwceennttyrtatiimoen-smaotfe0d, f3,em1a0l,eoCrr3l0:CmgD/k(gS/Dd)aIyGoSfBteRstrsatusbswtearnecedoosneddabyys o4r-a1l0,ga4v-a15g,e oornc4e-2d1aiolfy at gweesitgahttisonwe(Gr)e,roercofrrdoemddoanyt4hGe dtaoydaafyte2r1raptossatrprairvteudma(nPdP)d.ailClyiunnitcialltohbeseenrvdaotfiotnhse astnuddyb.ody On days 10, 15, and visible implantation 21G, sites, 5 rats per group the intrauterine per time location opfoienatchweermebreyuot/hfaentiuzseda.ndFoirmpelaacnhtafteimoanlteypwei:th wapapsrorxeicomradteedl.y 2Mahtoeumrsal(+vi3s0cemrianuwteerse)epxoasmti-dnoesdegarnodsswleyraendprbolcoeosdsesdamtoploebstawinerpelacoslmlaecstaemdples. cEomlblrecytoesd woenrdeaycosll1e5catenddo2n1Gd.ayF1e0tGu,seasnwdearmenieouttihcanfilzueidd,apnldacfeenttaalsb,laonoddesmabmrpyloess/fweetruesecsolwleercteed on day 2IG. `dTehleivlearsta5ndanniumraslesthpeeirr glritoeurps.weOrne ddaeyssig0n,a3t,e7d,fo1r4,thaendla2ct1aPtPi,onoaflfesvparliunagtwioenreanidndwievirdeuaallllyohwaenddtloed awnedigehxeadmi(sneexdesfosrepaabrantoer).malObnehdaavyiso3r,a7,nd1a4,ppaenadra2n1ceP.P, Ldiavmespwupesreinanceascthheltiitzeerdwaenrde cmiolukntaenddand b(4l:o3o0d msianmuptleess) wpeors-edcooslel.ecteRda.ndMoamtleymasellebcltoeoddpsuapmspwleesrewweeriegchoeldl,ecetuetdhaapnpirzoexdiamnadtgellyoo2dhsoaumrsples were collected on days 3,7, 14, and 21PP. . Psulpaesrmnaa,tamnitlsk,waemrneioatniaclyfzleudidbeyxthriagcht,paenrfdotrimsasuneceholmiqougiedncahtreom(paltaocgerntaap,hyem~brmyaos,sasnpdecftertousm)etry (HPLC-MS). Clinical Observations and Mortality There were no test substance-related clinical observations in dams or liters. * There was no mortality in mateal rats at any level tested. Body Weight and Body Weight Gain Mean body weight gain at 30 mg/kg/day similar to control during lactation. was lower than control during gestation and was Mean daily body and lactation. weights at 30 mg/kg/day were lower than control throughout gestation Mean body weights and lactation were similar to body weight gain the control group. at 3 and 10 mg/kg/day during gestation and --_-- PFOA: Lacon snd Place Transport Pharmacokinet Sayin Rats DuPont 13209 Reproductive Data + All animals were pregnant at scheduled sacrifice. + All dams delivereda livelier Tghreoupnsumtbhaetrwoefreimspalcarnitfaitcieodnonsietist,herresdoarypt1i0o,n,15a,nodr i21vG.fetuses were comparable across Thvearriaebiwlietryein2istmealrlsilzietearnsdinthtehesm3a0llmgn/ukmgb/edrayoflalcittaetrisonevgarlouuapt.edD(u5epetor tghreouipn)titnhseic relationship of this finding to tet substance administration is equivocal: + Pup survival an pup weights during lactation were comparable across groups Biochemical Toxicology + T1h1e.2c,o2n6c.e,ntarnadti6o6n.o6fpPgF/mOLAaitn3m,a1t0e,maanldp3l0asmmakappdeoasreedle0velbse, ratesspteecatdiyveslayt)eo(vmreatnhes of range of time points sampled. + Tanhde 3m3e.a1nucgo/nmcelnattra3t,i1o0n,saonfdP3F0OmAglinkgfedtaolsepllaesvemlas,corlelsepcetcetdivoenlyday 21G were 5.9, 14.5, + 3P,u1p0,plaansdma30comncken,trarteisopnecdteicvreelay)setdo fdraoymTdPaPy(30P7P, 2(.m8e,aannsdo4.f9 2g.9, m5.9a, tand3,1120.,0 pangd/mL at 30 mg/kg/day, respectively, and were similar on days 7. 14, and 21PP at alWage levels, + "aT3h,e c1o0n,caenntdra3t0iomng/okfgPFdoOsAe lienvemlisl,krwesapsecattisvetleya)dyfrstoamted(amyePanPs otofda1., y22.18P,Paantda6ll.2dousge/L. levels + d"Tahye 2co1nGce(nmteraatnisoonfof1.P5,F3O.8A, ainndam8i.1oriigcmflLuidatw3a,s1a0p,parnodxi3m0atmegl/ykgf,ourrestpiemcetsivheilgyh)erthoann on day 156 (means of 06, 07, and 1.7 jg /mL at 3, 10, and 30 mg/kg, respectively), T(hmeeacnosncoefn3tr.a6t,i9o.n4,oafnPdF24O.A4 uingpmlaLcenatta3,wa1s0,aapnpdro3x0immagt/eklgy,trweospteicmteivsehliy)ghtehranonondadyay21G 15G (means of 2.2, 5.1, and 13.2 ug mL at 3, 10, and 30 mak, respeciively). Th14e,3d3a,y a1n0dG1e2m.b5rgyo/mhaLd tathe3,hi1g0h,esatndco3n0cemngt/rkagt,iornesopfePctFivOeAly)pearngdrtahm odfayti2ss1uGe f(emteuasnhsaodf srelsipgehcttliyvelloyw)e.r lTehveeldsa(yme1a5nGseomfb1r.y3,o2h6adaandlo8w.e8rgt/ismsuLe.coantc3e,nt1r0a,tainodn 3t0hamn ikss,ue from rdeasype1ct0iGvoelryd).ay 21G (means of0.2, 0.5, and 1.2 ig fm. at 3, 10,and 30 mg, a PFOA: Lacttional and Placental Transport Pharmacokinetic Sudy in Rats Dupont13309 wIneicgohntclaunsdiowne,iugnhdtegraitnh)e caton3d0itmigo/nksgo/fdatyh.is Tsthuedrye, wtheerree 2wassmamlaltleirtntearls tionxtihceit3y0(rmegd/ukcge/ddbaoydy lactation `maternal group, a finding that was plasma and milk were at possibly test substance-related. steady state during the sampling Concentrations of PFOA interval. Steady state in concentration in milk was approximately ten times less than the steady state concentration in matemal plasma. The concentration half the steady state concentration in of PFOA matemal in fetal plasma. plasma on The milk day 21G was approximately concentrations appeared to be gfernoermadlalyy 3cPomPptaordaabyleTPtoP,thaencdonwceernetrsaitmiiolnasr ionnpduapyspl7a,sm14a,.anPdup21pPlPasamtaalclondcoesnetrlaevteilos.ns dPeFcOreAased was and detected in placenta 15G), and fetus (day (days 21G). 15 and 21G), amniotic fluid (days 15 and 21G), embryo (days 10 NN. --_-- /3 PFOA: Lactaions and Placemal Transport Pharmacokinetic Studiyn Rats DuPont13309 INTRODUCTION "pThhaermsuabcjoekcitnetetsitcssutbusdtiaenscei,n aadmumltonraitsu,mwhPoFlOeAb,oidsyaesluirmfiancattainotnuosfedthientiensdtussutrbisatlanpcreocweasssesf.ouInnd to tbreanmsupcorht mcoonrceenrtarpaitdiionnsfoefmatlheestceostmpsuabrsetdantcoemwahleesn faodlmlionwiisntgeroeradltodopsriengg.nanAtvafielmaablleesdiastlaiomnitethde and there is no from lactating existing data females.) on the concentration of the est substance transported via the milk OBJECTIVE `mTihlekoabnjedctbilvoeodofplthaissmsat,updlyacweanstat,oadmentieortmiicneflutihde,ceomnbcreyntor,atfeitouns,ofputph,e atensdt sfeutbasltaanndcepiunpmpaltaesmmaal following lactation. repeated oral dosing of the dam using 3 graded dose levels during gestation and SPONSOR AND TEST FACILITY `aTnhdisCsotmupdaynwya,s Wciol-msipnogntsoonr,edDeblya3waMreC.omTphaenysp,onSts.orP'asula,ppMrionvnaelsowtaas, eafnfdecEt.iLv.e dtuhePdoantte dteheNsepmoonusrosr aHuetahlotrhiazenddtEhneviwroornkmeonnttahleSccoinetnraccets., ET.1h.edsutuPdoyntwadse cNoenmdouucrtesdaantdDCuoPmopnatnHya,sNkeelwlaNrgbkoDrealtaowrayrfeo.r STUDY DESIGN A. Experimental Design -- Test Formulation Dose Concentration Time-Mated Group 1 _(mg/kg/day)* 0 mg/mL)" 0 Females 20 1 3 m 10 06 20 20 0 vFormulatio3ns0of test whoanee 1 detoni6z0ed water (NANOpure Gion2i0cd vier) b Twoearechaidemvinsisthteesreedcoonncceentdraaitliyonbsyoofraacltigvaevaignegraetddieonst,inthgevfoorlmuumleatoiofnsLwIekree. "adTjhuestceondrfoolrgsraomuppleapnuirimtay (r%e)c.eived NANOpure deionizedwater only at 5 mi. ------= PFOA: Lactation and Placental Transport Pharmacokinetic Study in Rs DuPont 13309 Dose Group Subsets And Dosing Schedules Subset NAunmibmearlosf ~~ Dosin(dgaPyse)riod" A 5 4-106 B 5 4156 c 5 421G D 5 4G21PP 3b GAengiemsatlastwoenr;ePrPa=npdoosmilpyaratsusmigned cach subse. Termin(adlaySsa)crifice 10G 156 216 21pp B. Selection of Dose Levels Dose levels selected for this study were based upon the resultsof a two-generation reproduction s3t0umdyg/ikngr/adtasy."foGrraopupprsooxifm3a0temlayl7e0adnadys30beffeomraelceohraatbsitwaetiroen.dosDeodsibnygoorafltgheavPaygefeamta0l,es1,3, 10, or ctohenrteinwuaesdntohrtoeustghsouubtstmaantcien-gre,lgaetsetdatmiaotne,rananld tloaxcitcaittiyonobosrearpvperdoxaitmdaotseelsyu1p12tod3a0ysmgto/tkalg./dWahy,ilae slight increase in offspring mortality, attributed to failureto weanlings at 30 mg/kg/day. thrive, was observed in Fy generation. MATERIALS AND METHODS NN* ANALYTICAL A. Vehicle The test formulation vehicle was NANOpure deionized water. B. Test Substance 1 Identification The test substance was obtained from 3M, St. Paul, Minnesota, and was assigned Haskell cLoanbtoarmaitnoarnytNs,umsybneornyPmFsO,Ahauzpaordns,reacenidpt.hazAavradioluasblmeatienrfioarlmacltaisosnifoincatthieopnusr)itwya,scopmrpoovsiidteidobny, the vendor and documented in the study records and report. 2 Puity `The vendor reported purity was 95.2% - 97.99% -- ra -- -- PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPore-13309 3. Test Substance Stability `The stability of the test substance over the courseofthe study was confirmed by purity analyses conducted near the beginning and end of the study. 4. Test Substance Administration, Preparation, and Sampling a Administration A`TnhiemtaelstssausbssitgannecdetwoagsesatdamtiinonisstuebrseedtsbyA,orBa,l agnadvaCg,ewoenrceeddoasielydaftora 7d,os1e2,vaonldum1e8 dofay5smLkg. respectively. Animals assigned to the lactation subset (D) were dosed for 40 days. Dose volumes were adjusted daily based on body weights. b. Preparation Test formulations of the test substance in the vehicle were prepared daily and stored at room temperature until used. in the study records. The method of mixing the test substance with the vehicle is documented c. Sampling : Test formulations at concentrationsof 0.6, 2.0, and 6.0 mg/mL of PFOA were prepared and collected for uniformity of mixing/concentration verification and stability mays Dbgur room temperature) on July 11,2003. On July 31, 2003 and August 21, 2003, test formulations at all levels were collected for concentration verification analysis. In addition, 0 mg/mL (control) samples were submitted for analysis with each set of samples. Samples submitted for analysis were analyzed the day they were receivedand/orwhen reanalysis was indicated. 5. Analytical Methods a Recovery Sample Analysis Concurrent with test formulations analyses, recovery of PFOA from spiked vehicle (NANOpure deionized high level water) was tested at the (6.0 mg/mL) to confirm low the level (0.6 analytical mg/mL), method. at A the mid level (2.0 mg/mL) and stock solution of PFOA was at the prepared in NANOpure deionized water. For all concentration levels, an appropriate aliquot of 1th0i1s0s0olmutLioonftNoAobNtOapinur0.e3 dmegio(nliozwe)d,w|atmerg. (mAildl),reacnodve3rymgsa(mhpilgehs) woefrtehethteesnt msiubxsetdanfcoredwiasspeardsdioend of the test substance in the vehicle. The samples were then processed and analyzed in the same `manner as the dosing samples at similar concentrations. b. Dosing Test Formulation Treatment Each dosing sample (0.5 mL) was diluted to 100 mL with NANOpure deionized water and mixed. The dosing samples were further diluted with NANOpure deionized water to an expected i -- PFOA: Lactational and Placenta Transport Pharmacokinetic Sudy in Ras DuPont13309 concentration of approximately 0.000003 mg/mL (50 pL injections) or 0.00003 mg/mL (5 uL injections) prior to analysis. sample initial dilution) was Before all final dilutions, added to each test sample the intemal standard to give an equivalent and the 0 mg/m. final concentration of the intemal standard and the matrix in all samples. Samples submitted indicated. for analysis were analyzed on the day of receipt and/or when re-analysis was Chromatographic Conditions LC parameters Tnstrument Column: HZeowrlbeattxPaRcXk-arCdS,Mo2d.1elm1m10x0 1H5P0LmCm, Flow Rate: 0S.u4mmL/min Injection Volume: Sor SOUL Column Temperature: 30C Column Switch: 4.0 minutes Mobile Phase: 0.15% Acetic Acid/Acetonitrile Gradient Tim0e0(min) % Ac5etonitrile 09 5 10 80 \N. 60 80 61 5 70 5 MS parameters IIonnsitzrautmieonntm:ode: Capillary voltage: Cone Voluage: Source Temperature: Desolvation Temperature: Scan function EMlieccrtoromsapsrsayQu(aEtStDr,onMeigcartoivTeainondem MS 27kV 15v 0c 350C 4p1er3fmlluzo(rpoaorcetnatn)oitco 3ac6i9dm(zPFO(dAa)u:ghter) [1.2:6i-"C] PFOA: 415 nz (parent) to 369 mz (daughter) Rinejteecntitoinosn).timeof PFOA and [1,2-di-"C] PFOA: approximately 4.9 to 5.2 min (5 or 50 uL. d. Calibration and Quantitation A stock solution of the PFOA (separate sample used as analytical reference) was made in NdeAiNoOnipzuerdweatdeeirotnoizmeadkweatcearl.ibrAaptpiroonpsrtiaantdearadlisqtuhoatts borfatchkeetsetdoctkhewetarregedtilcuotnecdenwtirtahtiNoAnNoOfptuhere diluted sample solutions. Before these aliquots were brought to volume, an appropriate amount of intemal standard ({1,2-di-""C) PFOA) was added. i PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont13309 (AnLaCl-yMsSi/sMoSf)P.FONeAgawtaisveb-yiohnigehl-ecpterrofsoprrmaaynwcaesliuqsueidd ctohrgoemneartaotgeranpeghayt/itvaenldyecmhamragsesdsipoencstorfomPeFtOrAy. `The ions were selected with the first MS quadrupole, collisionally dissociated using argon, and a ifnrjaegcmteinontsioofn twhaessamomnpilteorseodl.uti[o1n,s2-adnid-"c*aCl]ibPraFtOioAn sistaunsdeadrdass saonluitnitoenmsalwesrtaendmaardd.e aTnridplpiecaakteareas `were calculated electronically. `The calibration curve was generated by regression analysis using the peak area ratio from the PFOA and the intemal standard. Data for test formulations were compared to the calibration curves to evaluate the concentrations of PFOA. `(TCe.sVt.su=bssttaanncdearudndiefvoiramtitiyoni/nmtehaenvxeh1i0c0l)e owfatsheevmaelauastuerdedbycocnaclecnutlraattinigontsheofcotehfefidcuipelnitcaotfevsaarmipalteiso.n (uniformity of mixing/concentration verification)for each dosing level. A coefficient of variation less than or equal to 10% is the standard criterion at Haskell Laboratory for acceptable distribution of the test substance throughout the solution. `The mean result 0 determine the of the concentration concentration of the verification samples (n = 2) for each dosing level test substance for the respective dosing levels. was used Stability was evaluated by using the mean result of the uniformity of mixing/concentration verification samples a the baseline for comparing the corresponding stability resuls ~ IN-LIFE A. Test Species "The rat was selected for this study as it is a preferred species for reproductive toxicity studies and s`twraasinthweasspesceileesctuesdebdaisneda oprnecvoinosuisst2e-ngtelnyeraactcieopntarbelperohdeuacltthiosntasttuusdayn.d" oTnheextCernls:iCveDe(xSpeDr)iIeGnSceBR with this strain at Haskell Laboratory. JEuilgyht1y1n,u2l0l0i3par(o2u0s,fetmiamlee-mraattse)dfrfoemmaClheasrwleesreRirveeceriLvaebdoroantoJruileys8,,I2nc0.0,3Ra(l6e0igfhe,maNloertrhatsC)araonldina, with an assigned birth date of May 5, 2003. The rats for this study were approximately 63 days old and were at supplied by the day 1G vendor upon arrival." Body and are documented weights on the day rats in the study records were mated (day 0G) were B. Animal Husbandry I. Identification sEuapcphlireart pwraisoratsosisghinpepdiangu.niUqpueonnuremcbeeiprt,anedacihdeanntiimfiaeldwwaisthasasniAgnVeIdDa MHiacskreolclhiapniimmapllannutmbbeyrt.he Both the Haskell animal number and unique vendor animal number were recorded on cach cage [EE -- PFOA: Lactational and Placental Transport Pharmacokinetic Sudy in Rats DuPont.13309 cuanridq.ueAAmVaIsDterMilcisrtoocfhiupniiqmupelavnetndnourmnbuemrbsearrse,mHaaisnktealilneadniwmiatlh ntuhmebsetrusd,yarnecdorcdosr.responding 2. Environmental Conditions Aanndimmaalinrtoaoimnsedwaetraenmaacicnetpatianbelde artelaantiavcecheupmtiabdlietyteomfpe3r0a%t-ur7e0o%f(1t8ar-g2e6teCd (atta4r0ge%t-e6d0a%t)2.2-A2n4imCa)l rooms were artificially illuminated (fluorescent light) on a 12-hour light/dark cycle (approximately 0600-1300 hours). 3. Housing All rats were until sacrifice housed or day individually in 19G. Females stainless selected fsotreetlh,ewliarceta-tmieosnhsucbasgeets,(sDu)swpeernedehdouasbeodveincageboard polycarbonate pans containing bedding material on day 20G through day 21PP. 4. Feedand Water All rats were ad libitum. fed pelleted PMI Nutrition International, LLC Certified Rodent LabDiet 5002 All rats were provided with tap water from United Water Delaware ad libitum, A 5. Health Monitoring Program AfosllsopweciinfgipedroicnetdhuereHsasakreelpleLrafboorrmaetdorpyerainoidimcaalllhyeatlothenasnudreentvhaitrocnomnetnatmailnamnotniltevoerlisngarperboeglroawm, the. those that would be expected to impact the scientific integrity of the study; Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead, and other contaminants. + Feed samples are analyzed for total bacterial, spore and fungal counts. + Samples from freshly by the cagewashers. washed cages and cage racks are analyzed to ensure adequate sanitation rCeerqtuiifrieemdeanntismaanldfneoetd tiso uesxecde,egdusatraatnetdemedaxbiymtuhemmcaonnucfeancttruarteiorntsoomfeeketyscpoecnitfaimeidnannuttrsi,tiionncalluding sppreecsiefniceed ohfeathveyse:mectoanlts,amaifnlaatnotxsinb,eclholwortihneamteadxhiymdruomcacrobnocnesn,traantdioonrgstaantoepdhobsyphthaetemsa.nuTfahceturer would not be expected to impact the integrity of the study. "lTahbeoraantiomraylahneiamlatlh vaentderiennavriiarno.nmeEnvtaalluamtoinointoofritnhgesperdoagtraadmiidsnaodtmiinndiisctaetreedanbyyctohnediattitoennsditnhgat affected the validity of the study. is -- -- PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont13309 C. Quarantine and Pretest Period Rats were quarantined accordingto procedures outlined in Haskell Laboratory SOP LAOO3-P, and then designee. releasedfor the study upon approval of the Laboratory Animal Veterinarian or a D. Assignment to Groups ~ Randomization eUxppoenriamrernitvaall,gdraomuspswebryerraanndkoemd soanmpthleinbgasfirsoomftdhaeyr0aGnkbeoddlystw.eiTghhetsdiasntrdibaustsiiognnerdestuoltceodntirnomleaannd body weights for all groups that were not statistically different (p > 0.0). were then randomly assigned to each subset (A,B,C or D). Animals in cach group. E. In-life Observations 1. Clinical Observations Clinical observations were recorded on the day after arrival and daily until the end of the study. 2. Body Weights . Body weights were recorded on the dayafter rival and daly unl the end of the ud. 3. Lactation Procedures - Subset D N `The day when delivery was period (days 0, 3,7, 14, and complete was designated day 0 postpartum. 21PP), offspring were individually handled At cach examination and examined for paubpnsorimnaclabcehhlaivtieorrwaenrde acpopuenatreadncbey;saenxyadsesaodonoraasbpnoosrsmiabllepaufptesrwdeerlieverreycowradesdc.omLpilveetaendanddealidtter weights were recorded. Dead pups were discarded. F. Terminal Sample Collection and Sacrifice I. Gestation Subsets A, B, and C On by days 10, 15, and 21G, animals carbon dioxide inhalation. For assigned to Subset A, B, each female with visible and C, respectively, were cuthanized implantation sites, the intrauterine gTorcoastsieonxoafmienaacthioenmborfymoa/tfeetmuaslavnidsciemrpalwanatsatdioonnetaynped (blliovoedorsaemarpllyesrefsroropmticoanc)hwdearem rweecroerded. A ocobltlaeicntpedlfarsmoamstahmeplveesn.a cOanvadaaypp2r1oGx,imfaetteulsyes2wheoruerseu(t=ha3n0izmeidnuwtietsh)apnosotv-edrodsoesaenidnjpercotcieonss(eid.p.t)oof sodium carotid pentobarbital artery. and fetal blood samples were taken from a transverse cut made to the ES -- -- PFOA: Lactationsl and Placental Transport Pharmacokinetic Sty in Rats DuPont.13309 Gestation Subset Sample Collection and Sacrifice Schedule Terminal -- Subset Sacrifice Samples Collected A" 10G Matemal Blood," Whole Embryos" B 15G Maternal Blood," Amniotic Fluid, Placentas," Whole Embryos" 3 cMaternal b2lo1oGdsamplMeast(evronlaulmeB0l.o5omd.L"peArmanniimoatli)cinFulbuieds."coPnlaaincienngthae,p"arWinh,owlereecFoeltluescetse,d"2FheotuarlsBlood' b A(l2l3e0mbmirnyuotse,s)feptousse-sd,oasned; phleacrenttiasssuweesrweeproeocloeldlebcytelderas.oon 35 posible after maternal blood collection. a FAemtnailobtliocodFlsuiadmpslaempmlienmiimnuimmvuomlvuomleumaepparpopxrimoaxtiemlayte1l0y0 1L0.0c4olLlewcatsedpoinolteodbebsyciotnetra.iningheparinwas pooled by ler 2. Lactation Subset D a. Anesthesia Procedure Dams were removed from their litters approximately 1-2 hours before blood and milk samples were collected. Dams were placed in charged induction chambers (FLUOVAC anesthesia machine) and were anesthetized by isofluorane inhalation. When the dams appeared to be unconscious from the anesthesia, they were removed from the induction chambers and were checked for response to toe pinch. wAfetreermsoanmiptloersedwewrheilceolrleecctoevde,ridnagmfsrowmeraenersetthuersniead. tDoahmolsdiwnegrceargeetsuonretdhetiorthheoimreliPtehras w3fhteerr they recovery from anesthesia b. Blood Sample Collection `Sample Collection -- Subset D `Number of Pups for Pup Day EQ Maternal Samples' Milk Blood" Terminal Sacrifice > per litter Samples Blood PP Milk, Blood" 1 per litter Blood 14PP. Milk, Blood" 1 per litter Blood 5 21SPaPmp'les were collMeictlekd.'2 hBoluorso(d= 30 mimes)pos2tdposeer:loitthteerr samples were collBelctoeodd5"5 000 5 b Lpoistseirbsliezeafpteerrmmitaintg.e blood collection MMaatteerrnnaall bmliolokdsasmspmlpelse(vmoilnuimmeu0m.5vmoLl.umperaappnriomxaimcaotlelleycte1d00iuLb).es contining heparin. MPautperbnlaolodTesrammipnlaelmSianciimfucme volume approximately 100AL collcte in tubes containing heparin. -- W -- PFOA: Lacttional and Placental Transport Pharmacokinetic Sudy in Rats DuPont.13309 Maternal Blood samples were collected from the orbital sinus (days 3, 7, and 14 PP) or vena cava (day 21PP) approximately 2 hours (+ 30 minutes) post-dose. Fetal Randomly selected pups from these dams were weighed and euthanized on the same postpartum day with an overdose injection (i.p. of sodium pentobarbital. Pup blood samples were taken from a transverse cut made to the carotid artery on days 3 and 7PP or from the vena cava on days 14 and 21PP. Matemal and pup blood samples were processed to obiain plasma samples. Milk Sample Collection Milk samples were collected after blood sample collection. Oxytocin was administered by intraperitoneal injection to promote milk ejection from the mammary gland. Approximately 2 to S minutes after injection, the dam's nipple area was washed with warm saline or water. Milk awnads wceolrleecrteepdeaftreodmamsonreecestshaarny.oneMimlakmwmaasrcyolglleacntde;dmbaymamppalryyingglparnedsssuwreeretocthhaenegnetdirferebqauseenotlfythe gland, pushing milk toward the nipple and into a test tube. Pups were kept in polycarbonate pans with bedding during the milk collection prosgdure. On dhoaty w3aPtPerlpiattder,s hweearteedkegpetl pwaacrkmsd,uorrinhgeatthleamapb.sence of the dam by the use of either acleulating 4 Euthanasia After milk collection but before recovery from anesthesia, dams were euthanized by exsanguination. The remaining pups were euthanized with an overdose injection (i.p.) of sodium pentobarbital. Dam and pup carcasses were discarded. BIOCHEMICAL TOXICOLOGY SAMPLE ANALYSES A. Tissue and Plasma Analysis 1. Amniotic Fluid, Milk, and Plasma Plasma, milk, and amniotic fluid samples (pooled by liter), were processed by protein Lpraekceiwpiotoadti,onCO()P.PTA) u0s.i5nuggI/somlLutesoAlrutriaoynporfotpeeirnflprueocriopniotnaatinooniccoalciudmn(sAl(dJroincehsCChhermoicmaaltso,graphy, Milwaukee, WI) in acetonitrile (ACN) was used as an intemal standard. Samples were thawed and a 20 pL aliquot of each sample was applied to the PPT array. Samples were precipitated by adding appropriate dilution rate volumes of ACN/internal standard solution. Dilution rates sroalnugtiinogn)fwreorme1:u4ti(l6iz0eduLtoofcapitnutreematlhestsaanmdpalred csoolnucteinotnr)attioon1s:8w0i(th1i5n80thueLraonfgientoefrntahle ssttaannddaarrdd a ~ PFOA: Lactation and Placenta Transport Pharmacokinetic Study in Rats DuPon-13309 curve concentrations. The array was slowly eluted under vacuum into a 96-well receiver plate, centrifuged at ~3000 rpm for 10 minutes, and the extracts were analyzed by high performance liquid chromatography ~ mass spectrometry (HPLC-MS). 2. Placenta and Embryo Placenta samples (pooled by litter) were coarsely chopped using scissors; embryo samples (pooled by litter) did not require this step. Aliquots (approximate0l.5y g) of the embryo samples or of the placenta coarse homogenate were transferred to another tube and homogenized further using a Branson ultrasonic probe sonifier in the presence of ACN in a ratio of approximately 4:1 (v:w) ACN:tissue. The homogenized sample was then centrifuged at ~3000 rpm for 10 minutes. The supernatant was removed and taken to dryness by nitrogen convection, dissolved in intemal standard solution (0.5 g/mL perfluorononanoic acid in ACN), and analyzed by HPLC-MS. 3. Fes Fetus samples were pooled by litter and homogenized using a Waring blender in the presence of liquid nitrogen. Aliquots (approximately 0.5 g) of the homogenized samples were then transferred to another tube and homogenized further using a Branson ultrasonic probe sonifier in the presence of ACN in a ratio of approximately 4:1 (v:w) ACN:tissue. The homogenized sample was then centrifuged at ~3000 rpm for 10 minutes. The supematant was goed nd taken to dryness by nitrogen convection, dissolved in internal standard solution (0.3 pg/mL perfluorononanoic acid in ACN), and analyzed by HPLC-MS. 4. High performance liquid chromatography~ mass spectrometry (HPLC-MS) a Chromatographic Method CMoeltuhmond: CMoobliulmenpehmaspeesr:ature: P`Wlaatsemras cXonicenetMrSartiCor1n8a,an2al.y1si3s0mm25,pm AA:mbi5e0nmtM Ammonium Acetate B: ACN Gradient [mewn ws] [oo Te os70 [ho0 o| [oe10 Ce 0 1] SFtloopwtriamee:: Injection volume: 012150 mmiUnmin SAL = -- PFOA: Lactrional and Placenta Transport Pharmacokinetic Study in Rats DuPon 13309 b. Mass Spectrometer Method System: DMeotdeec:tor SourceL:M 1 resolution: HToMn enerregsoylution CEonltrlainscoen: ExLiMt2 resolution: HonMe2nerreasoyl2ution CMaapliilpllaireyr((KVV)): CExotnreac(tVo)r (V): RSoFurlcenest(eVm)p:erature: Desolvation temperature: MSMMoedtheo:d: cThimte: cn: Limitof Quantiation Wheaatteerr,sa2n7d9a0uLtioqsuaimdplCehrromatograph, equipped with quaternary pump, column MQuualtitprloeMRiecarcotiMoan sMoSnpietcotrrionmge(teMrRM) N1e0g0ative Electrospray 11000 52 51 210 62500 a0is 0130C 350C MRM, 2 wansiions ". 40-1131.0n0in 369.00 N DCuoellslon energy 01255656 4D6u9e.l0:0 419.00 025 5ec C0.o0l5lipsgo/nmLe.nergy: 15ev STATISTICAL ANALYSES For all quantitative data presented in the report, descriptive statistics were performed that included calculation of mean and standard deviation. --_-- PFOA: Lacttional and Placental Transport Pharmacokinetic Study in Rats DuPont13309 RESULTS AND CONCLUSIONS AY w -- PFOA: Lactariona and Placental Transport Pharmacokinetic Sty in Rats DuPont 13309 ANALYTICAL DATA A. Test Substance Stability Analyses `Samples of the test substance were analyzed near the beginning and end of the study. These analyses indicated that PFOA was stable over the course of the study. The average of the active ingredient was 91.9 = 1.0% and 96.6 2.1% for samples analyzed July 14, 97.99% 2003 pure. and September The difference 4, 2003. between theThsepoPnsFoOrArewpaorstreedppourrtietdy bayndthtehespeoxnpseorrimteonbteal9d5a.t2a are most likely due to analytical variability. B. Test Substance Formulation Analyses (Table 1, Appendix A) I. Chromatography 4P.F9.O10A5e.l2utmeidnuftreosm(t5hepLHPorL5C0cuoLl.uimnnjeacstiaonrse)sfoolrvetdhepeneagkawtiivteh iaonr.eteRnetpiroensetnitmaetoifveapLpCr-oxMiSm/atMeSly chromatograms are shown in Appendix A, Figures 2 (a - 1). Test substance was not detected in tshtean0damrgd/imntLrocdountcreodlafdoertmeucltaatbiloen.pe[a1k,2i-ndit-h"e'cCo]ntpreorlfl(u0ormogoc/tmaLnosicamapclied)uisnedthaessatnuindteTrhnials was determined by comparisonof the NANOpure deionizedwater (diluent) used as a wash the LC/MS analysis and the NANOpure deionized water (diluent) with intemal standard along with the 0 mg/mL. sample (control) with intemal standard. Refer to Appendix A, Figures 2 (a -c). 2. Recovery Samples DvaertiaaiblielditaynoaflytthiecaalnarleystuilctaslofmertehcoovderwyassadmepmloensstarraetseudmbmyartihezecdoeifnfiAcpiepnetnsdoifxvAa,riTataibolneoIf. tThhee recovery results at each targeted dosing concentration (approximately 0.6, 2.0, 6.0 mg/mL) over 9th2e.c0o%urtsoe1o0f6t.h2e%sotufdyn.omiTnhael m(emaesaunrepderccoennctenrtercaotvieornys=of97P.F0 O+A8.f0or%,thCe.0V..6 =mg8%/)m.L Tlheveelmewaesruered concentrations of PFOA for the 2.0 mg/mL level were 83.0% to 100.2% of nominal (mean p6.e0rcmegnt/rmeLcolveevrely =we9r2e.690=.83.%8%to, C1.0V8..5=%9o%f).nomTihnealme(amseuarnedpecrocnecnetntrreactoiovnesr=oyf9P9F.2O+A9f.o1r%t,he C.V.= 9%). Based on this data, the analytical method performed satisfactorily over the entire concentration range for the study. 3. UniformityofMixing/Concentration and Stability Samples Aonfamliyxtiincga/lcroenscuelnttsrfartoimondovesriinfgicsaotliuotnioannsdcSo-lhloecutrerdooonmJutleymp1e1ra,t2u0r0e3satanbdilaintyalayrzeesdhfoowrnuniinformity Appendix A, Table IT and Summary Table 1 -- - 5 -- PFOA: Lactuiona and Placenta Transport Pharmacokinetic Sudy in Rts DuPon 13309 `sTthabeilfiotlylaonwailnygsetasblfeorstuhmismasraimzpelsintghedareysoulftsthfeorPhFoOmAogeprneepiatrya/ticoonn.centration verification and `SSaammplpelTeyDapy NmomyimnLal MemasgumreLd" % NMoeminnal C_V_(.%)_5SNuomibnal 11-uly2003 Uniformity Concentration 0 ND -- -- 060 0613,0558 20 221,207 977 1070 7 1020 5 1020 60 668,630 1082 3 1040 ab MSaemapnlerseshuelsdlo5 hotuhersaantalryosiosmoftetmhpeerdautpulriec.ate samples. cDoenntortoelscnauosneddeatedcetteedc.tabRleeppoerstkedforrehseusteastesbuabssteadoncne sinhotwheincognttrhoalt.thSehiontwenrnailn sFitgaunrdeasr2d a(ddaict)i.on othe The results for PFOA samples prepared and collected on July 11, 2003 indicated that the test sthuabnst1a0n)ceanwdasstaatblteheintatrhgeetveedhiccolnecwenhternathieolnds 5(=h8ou.r2s%aotfrnooommintaelm)p,eraadteuqrueatfeolryamllixleevedls(.CVTelsetss substance was not detected in the 0 mg/mL samples. 4. Concentration Verification Samples Analytical results from dosing solutions collected on July 31, 2003 and August 2182003 and Taanballeyze1.d for concentration verification are shown in Appendix A, Table II and Sumiary `The following sampling days table summarizes the results of the PFOA preparation. for concentration verification analyses for both PrepDaaration Nmommin.al Vimegw/emdL" %AvNeormaigneal o__v% 3hy200 060 20 0497,0493 172,169 825 1 555 1 21-Aug2003 60 0s 0458986.,0560173 150007 32 20 60 190,199 545.504 075 3 85 5 3b RDeupploirctaetderseasmpilseasr petrhelemveelanwoefredaunpalliyczaetde.e-Cs.aV.mpclailncguloaftetdheoorvgeirniafylusnuibfmoirtmtietdysoafmmpilxetsurbee.cause the original sReapmoprlteeadnarelsyuslitsswaarsenfortomacrceespatambplleindgueof(0tshtaornidgairndalcusruvbempirteepadrastaimopn.les and dition (0ahigher level for aprneaplayrsaitsio(nS.ul. nection) because th orginal sample analysis wasno acceptable dueto standard curve "The results for samples prepared and collected on July 31,2003 indicated that the test substance was at PFOA the targeted levels (+ 17.5% of nominal) and samples. Test substance was not detected in adequately mixed (CV the 0 mg/mL samples. less than 10) for all % PFOA: Lactationsl and Placental Transport PharmacokineticStudy in Rats DuPont13309 `sTuhbestraesnucletswafosrastatmhpeletasrpgreteepdarleedvealsnd( co1l2le.c5t%edofonnoAmuignuaslt) 2a1n,d2a0d0e3quiantdeilcyatmeidxtehdat(tChVe tleessts than 10) for all PFOA samples. Test substance was not detected in the 0 mg/mL samples. C. Analytical Conclusions Rseusbuslttasncferwomasthmeiaxneadlypsriospoerfltyhe(CPVFOleAssdtohsainng10s)o,luattitohnestdaurrgientgedthleevsetlusd(y i2nd0i%catoef tnhoamtitnhael)tesatnd stable under the conditions of the study. Test substance was not found in the 0 mg/mL samples. \,hN - 7 PPFFOOAA:: LLaaccttraitioonnaallaannddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiicc SStuuddyyiinnRRaattss_ DDuuPPoonntt-1133330099 REPRODUCTIVE TOXICITY EVALUATIONS IN-LIFE TOXICOLOGY A. Matemal Observations (Figures 1 and 2, Appendices B, C, and D) ``TThheerree wwearsennoomtoersttalsiutbystaatncaen-yrellevaetledtecsltiendi;caalllobasneirmvaaltsioonns.study survived to scheduled sacrifice. Means and standard deviationsfor body weight and body weight gain data are in Appendices C and D. Mean 34.5g body weight gain at 30 mg/kg/day was for the control group) and days 10-15G lower than (28.7 vs control 33.8g in during days the control 4-10G group). (24.9 These vs rgerdouucpti(o1n3s8.r1egsuvlsted15i2n .o7verfaolrlthweeicgohnttrgoalignrdouurpi)nfgogredsatyasti4o-n2t1hGa.t wMaesa1n0b%odloywweeritghhatngtahiencaotntrol 3H0owmegv/ekrg,/dmaeyanwadsaisliymibloadrytoweciognhtrtoslrdeumraiinngedlacatpaptriooxni(m1a1te.l2yv4s%7.l8owfeorr the control group). than control throughout ~~ * `gestation and lactation for this group. Body weights and during gestation and lactation were similar to the control body weight group. gain at 3 and 10 AN mg/kg/day Mean Body Weight Gain (grams) for Selected Intervals During Gestation Dose (my/kg/day) [GestationbaysT07 [90 7 | 3| us 21 364 29 828 1057 1049 793 1015 n8 31s 353 287 716 921 583 849 421 1527 1444 159.7 1381 1576 2.66 20.16 3.71 Data presented: M`eSatannsdard Deviation B. Reproductive Data 1. Gestation Subsets (Appendix E) All animals were pregnant at scheduled sacrifice. `The that number of implantation sites, were sacrificed on cither day resorptions, and live 10G, 15G, or 21G. fetuses were comparable across groups -- Tm PFOA: Lactstonsl and Placental Transport Pharmacokinetic Sty in Rats DuPon-13309 2. Lactation Subset (Appendices F, G, and H) All animals delivered a live litter. There were no test substance-related clinical observations in litters during lactation. Psuubpstsaunrcvei-vraellaatneddppuuppwmeoirgthatlsitdyudruirnignglaclatcattaitoinown.erTehecroempwaerraebl2esamcarlolsslitgtreoruspisn;tthhee3re0 wmags/kngo/dteasyt lactation group. Due to the intrinsic variability in liter size and the small numberoflitters evaluated (5 equivocal. per group) the relationship of this finding to test substance administration is Number of Pups Born for Individual Litters 0 Dose (mg/kg/day) 3 10 30 13 15 2 13 14 4 1 3 1 1 2 6 16 16 14 n 2 13 14 3 Mean 148 124 132 az Standard Deviation 27 483 110 449 BE -- PPFFOOAA:: LLaaccttraitoinonaallaannddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSwtuddyyiinnRRaattss ___ DDuuPPoont113330099 BIOCHEMICAL TOXICOLOGY A. Matemal, Fetal, and Pup Plasma (Tables 2-3, Figures 3-5, Appendix I) Ttihmeecpoonicnetnstrsaatmipolnedo.f P`TFhOe Ameiannmactonecmeanltrpaltaisonmsaaatpspteeaardeyds1t0atbeewaetrsete1a1d.2y,s2ta6t.e8,ovaenrdt6h6e.6rapngge/moLf at 3, 10, and 30 mg/kg dose levels, respectively. collected on day 21G were 5.9, 14.5, and 33.1 The mean concentrations of PFOA pg/mL at 3, 10, and 30 mg/kg dose in fetal levels, plasma r1e2s.p0ecptigve/lym.aLtP3u,p1p0,laasnmda3c0onmcge/nktgr,atrieosnpedcetcirveealsy)edtofdraoym day 3PP (meansof2.9, 5.9, and 7PP (0.7, 2.8, and 4.9 pg/mL at 3, 10, and 30 mg/kg/day, respectively), and were similar on days 7, 14, and 21PP at all dose levels. B. Milk (Table 4, Figure 6, Appendix 1) l`eTvheelsc.onTcehnetrmaetiaonncoofncPeFntOrAatiinonmsilatkswtaesadaytssttaeteadwyesrteate1.f1,ro2.m8,daaynd3P6.P2tpogd/amyL2a1tP3P, a1t0,allanddose 3th0emsgt/eakdgydsotsateelceovneclse,ntrreastpieoctnisvoelbys.erTvheedsiencmoantceenmtarlatpiloanssmaa.reTahppermoixlikmactoenlcyentternattiimoenss alepspseathraend 10 be generally comparable to the concentrations in pup plasma. hy C. Amniotic Fluid (Table, Figure 7, Appendix I) `The concentration (means of 1.5, 3.8, of PFOA in and 8.1 ug amniotic /mL at 3, f1l0,uiadnwdas30amppgr/okxgi,mraetsepleyctfiovuerlyt)imtehsanhiognhderayon15daGy(2m1eGans 0F0.6,07,and 17 ug /mL at 3, 10, and 30 mg/kg, respectively). D. Placenta (Table 6, Figure 8, Appendix I) `0T7h3e.6c,o9n.c4en,tarnadti2o4.n4 ofuPgFfOmALiant p3l,a1c0e,ntaandwa3s0ampgp/rkogx,imraetspeelcytitvweolyt)itmheasnhoinghdearyon15dGay(m2e1aGns(moefa2n.s2, 5.1,and 132 pg /mL at 3, 10, and 30 mg/kg, respectively). E. Embryo and Fetus (Table 7, Figure 9, Appendix I) `1T2h.e5dpagy/1m0lGatem3,br1y0,oahnadd3t0hemghi/gkhge,strecsopneccetnivterlayt)i,onanodftPhFeOdAayp2e1rGgrfaemtusofhtaidsssuleig(h1t.l4y, l3o.3w,eranldevels t(i1s:s3u,e2.c6o,ncaenndtr8a.t3igon/tmhaLnattis3s,ue10f,raonmdd3a0ym1g0/Gkgo,rrdeasype2ct1iGve()0..2T,h0e.5d,aaynd151G.2epmgb/rmyLo hatad3,a1l0,owaenrd 30 mg/kg, respectively). Eg PPFFOA::LLaacttioonnalsanndPPllaacceennttaalTTrraannsspportPPhhaarrmmaaccookkiinneetticSStuudyiinn RRaatsts_ DDuPoonntt113333009. CONCLUSIONS gUanidne)ratth3e0cmogn/diktgi/odnasy.ofTthhies msetuadny,ntuhmebreerwaosf pmuatpesrbnoarlntowxaiscitlyow(erredtuhcaendcbonotdryolweaitg3h0tmagn/dkgw/ediagyh,t amaftiendmianlg pthlaatswmaasancdonmsiildkerweedrpeosatsisbtleyadtyeststsautbesdtuarncien-gretlhaetseda.mplCionngceinnttreravtail.onsStoefaPdFy OstAatein cmoantceemnatrlatpiloansmian. mi`lThkewcaosncaepnptrroaxtiiomnatoeflyPFteOnAtiimnefsetlaelsspltahasnmathoensdtaeayd2y1stGatweacsonacpepnrtorxaitmiaotneilny `hgaelnfertahlelystceoamdpyasrtaabtelecotnocetnhtercaotnicoenntinramtaitoenrsnainl ppulapspmal.asmTah.ePmuiplkpcloanscmeantcroantcieonntsraatpipoenasrdeedctroeabseed fdertoemctdeadysin3ptloac7ePnPt,a a(dnadyswe1r5easnidmi2l1aGr)o,nadmanyisot7i,c14f,luaidnd(d2a1ysPP15ataanlldd2o1sGe),leevemlbsr.yPoF(dOaAyswa10sand 15G) and fetus (day 21G). RECORDS AND SAMPLE STORAGE SNpeewcairmke,nDse(liafwaaprpel,icoarblaet),IrronawModautnat,aainndRtehceofridnsalMraenpaorgtemwielnltb,eWrieltmaiinnegdtoatn,HaDseklealwlarLea.boratory, \,hN --_ , _---- _-- 3 _-- - -- s PFOA: Lacational and Placenta Transport Pharmacokinetic Study in Rats DuPont 13309 REFERENCES 1. DViasntdriebnutHieounv,eMle,taJ.bPo,liKsums,liakinsd,EBl.iLm,inVaatnioRnaoffelPgehreflmu,oMroJo.ctaannodiPceAtcerisdoni,n RM.alEe. (a1n9d91F)e.maTliessue Rats. J. Biochem. Toxicol. 6920, 83-92. 2. DuPont Haskell Laboratory (2003). Unpublished report, DuPont-7473. Perfluorooctanoic Acid: Toxicokinetics in the Rat. 3. rBeuptreondhucotfifv,e JtLo.xiKceonlnoegdyyo,fGa.Lm.m,oFnriaumem,pSe.rRf.l,uOor'oCoocntnaonroa,tJe.(aAnPdFYOo)rki,n Rt.heG.rat(.200F4r)o.m:The `rAerpgruosduLcatbioornatsotruideyso(f20a0m2)m.onOirualm(pgearvfalgueo)ro2o-cgteanneoraatteio(nA(PoFnOe)liitnerrapt.er generation) 4. DNuuPmobnetrH(aMskRe)ll56L7a2b.orUantopruybl(i2s0h0e2)d.ReDpuoPrto.ntDHuaPsoknetllHaLsakbeolrlatLoarbyoMreatdoircyalAnRaelsyetaicraclhRPerpojoerctt Number DuPont-13707 and DuPont. 13708. 5. HfoarzaAnradlyEsviasluaantdioEnvaDliuviastiioonn,oSftSaunbdcahrrdoEnviacluaantdioCnhrPornoiccedEuxrpe,osTuorxeicSittuydiPeotsenPtaiyanlt:er,GuOi.dEa.ncete al, United States Environmental Protection Agency, Office of Pesticide Programs, Washington, D.C., 20406. EPA-540/9-85-020. (June 1985). WW 6. Risk Assessment of Notified New Substances. EN), Chapter I, Sections 2.24 and 2.25. 1994 Technical Guidance Document (XI/283/94- 7 PPFFOOAA::LLacatcattaiioonnaaannddPlPascceenntaallTTrraannssppoorrtt PPhhaarrmmaccookkiinneetticcSSttuuddyyninoRtatss DDuupPoont-1133330059 TABLES NN - % PPFFOOAA::LaLcatcaitaitoonnaall aannddPPllaacceennttaalTTrraansportPPhhaarrmacokinetiiccSwtuddyyiinRRaats TABLES EXPLANATORY NOTES Abbreviations G= LOQ = <LOQ = NA = PP = SD. = Gesation Limitof Quantitation, 0.05 pg/mL. Less than limitof quantitation Notapplicable Postpartum Sundard Deviation DDuuPPoonntc.113333009 A wo PPEFOOAA::LaLcatcattaitioonnaallaannddPPllaacceennttaallTTrraannssppoorrtt PPhhaarrmmaaccookkiinneettiiccStuudyyinnRats __ DuPont1133099 TABLE | SUMMARY OF DOSING TEST FORMULATION ANALYSES Sample Type Dosing Concentrations and Stability of PFOA (mg/mL) Uniformity/Concentration Nominal 06 -- 20 60. July 11,2003 " 0613 1022) 221 (103) 668 any " 0558 207 630 Average Measured Cone." 030) (1033) (1050) 0.586 214 649 SAtvaenrdaagredPDeervcieanttioNno"minal 21090)4 10071.00) 0280.227) Coefficient of Variation" 7% so 4% hStoabuirlitRyoom Temperature 0612 204 624 (1020) 1020) 1040) JCuolnyce3n1t,r2a0t0i3on Verification " n AAvveerraaggee MPeeracseunrteNdomCionnca.l*" CSoteafnfdiaciredntDeovfiaVtairoina"tion 5024987) (1#620) 488 O\N 13) 0493 2) 1.69 43) 507 3) 0495 171 498 25) 20003 20052) 230.10)3 1% 1% 3% August 21,2003 ul 05% 93) 190 50) 5.5 08) n 10062123) 19939) 554004) AAvveerraaggee PMeeracseunrteNdoCmoinnca.l 0.604 (1007) 195 15 525 15 L_ o`CoStfeaffniidccairedntDoeovffiVaVataririoinaattiioonn*' 2 220%01 % 330%%06 5250%%29 ba TvNahuremibaatevireosrnaogifnedpmugepelanistcuharteeesdesscaomapnlcreesnt.ihreatrieosnp,ecativveerapgeeperrcoecfntenoofmninnotamlinvaallu(eisn. parentheses), standard deviton, and coefficient of vTahreiaatvioenraogfethmeeamseuarnedofcodnucpelnitcraatteioen-,ialvuetriaogneopfetrhceenrtioifnnaolmisnaamplle(si.n parentheses), standard deviation, and cocficient of Tvahreiaatvieornaogftehmeeamseuarendofcornec-ednlturtaitoinoonfatvheeroarggeipnearlcesnatmopflneosmtionhailg(h1epevnethfoersae)n, sitan(dSaLr.d ndeevcitaitoino)n., and coefficient of _-- 3 FPEFOOAA::LaLcacttsaitoionnaallaannddPPllaacceennttaallTTrraannssppoorrtt PPhhaarrmmaaccookkiinneettiiccSSttuuddyyiinnRRaattss DuDuPPoonntt-1133330099 TABLE 2 CONCENTRATION (ug/mL) OF PFOA IN MATERNAL PLASMA DURING GESTATION AND POSTPARTUM Day 51066 2IPG uTPPP 2PP AverSaDg.e -_-- "AveraOgemghgSD. <<900QQ NNAA <a00QQ NNAa 0<0QQ NNAA <0Q NA <NLOAQ Averag3me asSD. _ "omg Average SD. 1855932 1120966 moost 222m7 09609 0299%0 904 Lol 26420 H2aslo 24200 266m8 2308 22080 88 sis 12716 248214 Averagoe.mgSD 04 7955 894 su 6543360 1174.7866 65649615 I16s3 63 Lis 6965604 ~~ 3% PPFFOOAA::LLaacctttaitioonnasl!aannddPPllaacceennttaall TTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSStuudydin yRatis nRats___DDuuPPoonntte1133330099. TABLE3 (CONCENTRATION (g/mL) OF PFOA IN FETAL AND PUP PLASMA. Sample -- Day Omgke Average SD. Averam ge p SD. gAveroagemgSD. AverSagoewgisSD. Feal Pup 216 PP <0Q OQ Na NA S88 060 28 070 148 LSI 594 las 30 196 46 16 PPupp 1TPPPP O<0QQ NNAA 0067s 001200 22727 003s% 4a921 i12n8 rP--p ----2PP----<l--0Q ----NA ----1--28 ----om----3--25 ----0s:------ 736----21-- 7 \No 3 PFFOORA::LaLaccttaaitioonnasl!aannddPPllaaccenetnatlaTTrraannssppoorrtt PPhhaarrmmaaccookkiinneettiicc SSttuuddyy iinnRRaatsts_ TABLE 4 CONCENTRATION (ug/mL) OF PFOA IN MILK DuDPuPoonntt1133330099 Day -- "AveraOgemgigSD. AveragTe mgSkD.g Aveoragme gSD. Ave3ra0gemksSD. PPPP 0<0QQ NNAA 2uPpPp <0201Q NNAA 0794 00226 L[sRE0T06 227040 oosmt 33057 oLsst a9 576 120 126 64s 798 138 16 Average SD. NNAAY 010097 20680 611066 ma eAvereage t not cas lculated beeceauese3 of 4e values <LOQ.eeeerste A -_-- 3 ZPEFOOA::LLaaccttational andPPllaacceenntalTTrraansporrttPharmacokineticSSutdudyyininRRaatts DuuPPoonntt-13309 TABLE 5 CONCENTRATION (g/mL) OF PFOA IN AMNIOTIC FLUID T Day e Average p SD. e AveragSeegaSD. _ 21566 <00QQ NNAa -_ 016500 0036 Aver1a0gemehSD. 037706 0o8i1s Aver3a0gemgsSB. 10 813 ool 086 PFOA: Lacttional nd Placental Transport Pharmacokinetic Study in Rats DuPont.13309 TABLE 6 (CONCENTRATION (ug/g) OF PFOA IN PLACENTA Omg. Day Average SD. 156 2G <0Q <0Q Na Na Aver" age mSg D sAverao ge mSDg . 2m 1m 35 ost 510 170 937 17 Somgke Average SD. B23 23 an NN wo PFOA: Lactationsl and Placental Transport Pharmacokinetic Study inRats DuPont13309 TABL7E CONCENTRATION (g/g) OF PFOA IN EMBRYO/FETUS Tissue - Day EEmmbbyyoo 115066 Fews 21G Omg, Average SD. _ <<LLOOQQ NNAA <l0Q NA AveraSge mSg_D.h_eAveraTgeomgSDk.s 012440 00390 127 02% 03353 o08i1s 261 03 AverSageomgSDk. s 1122449 o35z0 sm 236 ~ oT PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont-13309 FIGURES NN a PFOA: Lactational and Placenta Transport Pharmacokinetic Study in Rats FIGURES EXPLANATORY NOTES Abbreviations G= LOQ = <LOQ = PP = Gestation Limitof Quantitati0o.n05, pg/mL. Less than limit of quantitation Posiparum DuPont13309 a -- ES 2 : 2 | g 2g Z gz \ IN \ _ oO) \ HFriil - 3 tet = leg # | 8 :7 :g 1i y\\ 1 = \k -| n& nn8on 3 =z 4S  ZF : 2 {4 la 2 Fa 5:44 = 3 Ld HHH g2 py lh LL 32 } a. 0: : = 88: 7 : nin tien g % PFOA: Lactations and Placenta Transport Pharmacokinetic Study in Rats Dupont. 13309 FIGURE 3 CONCENTRATION OF PFOA IN MATERNAL PLASMA DURING GESTATION AND POSTPARTUM [eee [ lIeC || | 50 | mn Ew, = 3mgk|g | %y + 10 mg/kg| & 40 3 30mig, . I 2wo01| AN " {|| o--_ | 0G 1G 21G 3p TPP 14PP 21PP | Day Note: Concentration in the 0 mg/kg group = <LOQ (0.05 pg/mL). -- -- wo PRON: Lion nd Pes spor PokiSteyisns Datos FIGURE 4 CONCENTRATION OF PFOA IN PUP PLASMA [3 || 124]i EL [EE [E7 R i 2 ol | ~~ 3 Tw 3PP TPP 14PP Post-Partum Day 1 | [mar] ---- 10 mg/kg | [2o0 mxy N || 21PP Note: Concentration in the 0 mg/kg group = <LOQ (0.05 g/mL). -_-- PFOA: Lactation! and Placenta] Transport Pharmacokinetic Study in Rats Dupont 13309 FIGURE 5 CONCENTRATION OF PFOA IN FETAL PLASMA ON GESTATION DAY 21 |0 | oss 0 Fas 8 51 Cl10 [oe 0 ma res 3 10 Dose level (mg/kg) || | || | ~ ER EE -- wo -- PFOA: Lacon and Pcs Tsphasan ityin ts. FIGURE 6 CONCENTRATION OF PFOA IN MILK Don 10 'sl | i | | 7o7f <5 44 3d [rams| = 10 mg/kg =30 mg/kgfi 21 | FiE --_-- | -- AN | 3pp PP 14PP 21PP Postpartum Day | ee -------------------------------------- Note: Concentration in the 0 mg/kg group = <LOQ (0.05 pg/mL). _-------- PPFROONA::LaLcacttaaitoinoanl!aannddPPllacaecnteaslTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSutdyyininRRatass _DDuuPPoonnt1313330099 FIGURE 7 CONCENTRATION OF PFOA IN AMNIOTIC FLUID ree boy 9 | 83] [326 2 EE 8a 5mais | 010 myke 30 mg/kg 34 | 24 | | 1 <Lo Q <LoQ NG 0 t-- rem |-- | 156 216 | Gestation Day | L tee -------- er --i ---------- Note: Concentration in the 0 mg/kg group = <LOQ (0.05 pg/mL) 5 PRO Lucan andPc Transpo Pharmacokinet Suyin as Dupo 13308 FIGURE 8 CONCENTRATION (ug/g) OF PFOA IN PLACENTA | 30.00 | 2500 20.00 > | | 2 15001 1& | | 10.00 || 5.00 | <L0Q <L0Q 000 4 -- Day 15 Day 21 | Gestation Day Note: Concentration in the 0 mg/kg group = <LOQ (0.05 pg/mL). | [ | (@3mgig | 010 mee | CE ~| | 3 | | | -------- PPOFAOA:: LLaaccttiaoinoanlaannddPPllaacceenntalTTrraansport PhaarrmmacokkiinneteitcSSutuddyy iinnRRatuss___ DDuubPoomnet1133330099 FIGURE CONCENTRATION (ug/g) OF PFOA IN EMBRYO/FETUS -- [tS |e 144| od 3 3 10 1 [&2) : 47 | o24alL0]Q, Day 10 <L0c Q el<L0Q Day 15 Day21 Gestation Day Note: Concentration in the 0 mg/kg group = <LOQ (0.05 g/mL) | JE [m3 mgig | B10 mee | m3my0 O"N i | --= PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont: 13309 APPENDICES N, No 53 PFOA: Lacttional and Placental Transport Pharmacokinetic Sudy in Rats DuPont-13309 APPENDIX A Ne ANALYTICAL DATA -- ES PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont-13309 Table I. Recovery ofPFOA AddedtoDosing Vehicle Sample `mg/ml PFOA Percent Type Nominal Measured Nominal Recovery Recovery Recovery" 0.584 0620 1062 0585 0542 26 0.587 0540 920 Mean: 97.0 +80, CV.8% Recovery RecoveryTM Recovery 195 1953 1002 195 1618 830 1.96 L854 946 Mean: 92.6 + 8.8, CV.9% Recovery 584 6336 1085 Recovery 5385 5283 903 Recovery 5.87 5799 088 Mean: 99.29.1, CV.9% \ (A) PprreopcaersesdedonwiJtuhlyun1i1,for2m0i0t3yofmixing/concentration verification samples from dosing. "Ss (B) () Processed Processed with with concentration concentration verification verification samples samples from rom dosing dosing prepared prepared on on July 31, 2003. August 21, 2003. 5 PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont.13309 "Table I. UniformityofMixing/Concentration Verification and Stability of PFOA in Dosing Solutions Sample Type Nominal `mg/mL PFOAMeasured Percent Nominal TI uly2003 Uniformity/Concentration CovroL 000 ND -- al 060 060 0613 0558 1022 930 Mean: 0.586.004 07.7%) al 20 2c2v1.r% 10s. n 20 Mean: 22.0174 20.10 (110073..05%) " 60 6c6v8.se 3 " 60 Mean: 66.3490027 105.0 (108.2%) cv. 5 Hour' 060 612 1020 20 200 1020 60 624 1040 (A) dDeetneoctteasblneopneeakdetoercttehde. esRtepsourbtsetdanecesainstaheecboantsreold.onShsohowwniinngFhait gthu2e irnatee.r.nsa.l sandard ation tothe come]y causead (B) vTahreiaatvieornaogfedumpelaiscuarteedscaomnpcleensi.ration, average percent of nominal (in parentheses), standsrd deviation, and coefficient of () Stabile samples held Shours oom temperature 5% PFOA: Lactaional nd Placenal Transpor Pharmacokinetic Sudy in Rats DuPont13309 "Table IPIreCpoanrcaetinotnraDtaioyn Verification ofPFOA imngD/omsLi.nPgFsOolAutions Percent 3Sa1m3p0l1e52T0y0p3e Nominal Messed Nominal ConcentraCtoinontrVeorilfication 00 No - " 060 0497 028 n 060 Mean: 0.044905320.003 2232 cv n" 2200 1162 584650 Mean: 1712002 55) cv. " " 6600 praiss) 854153 Mean: 4980.03 3.0 cv. 21-Aug2003 ConcentatCoonntVreorlification n n 00 No 060 0596 060 0613 Mean: 0c0v0.420%01 AN - * 93 1022 (1007) "" 2200 119990 999550 Mean: 1c9v5.2s0%06 07.9 nn 6600 ssoas 980480 Mean: 3c2v5.+s0n29 79) A)Tvharisvteoanogfedmoelascureedscaomnpcleen.iaion, verge pce of nomial (1 penises), sandr devision,andcocficient of () ) MdDeeetaencnttaseblunelopneefaokereaftnoraeltdyh.seesfsRteopsdoburspttleaidcnaceteesirleha-esracombnpatsioenld.fonhSsheohwornogiiwnnFaiilhgasuntrpehgsee 2.t3e-Om0a.rgssansndlysiisiofnsp0letsh cwoanstoncvasaedd ) Mdueeanoersolr finorsaananldysrescfuorrreepsrepaaramtiopnoflthiining samples and dilution to nlsofsamples was ot valued du err 1 sandnd curve preparion higher evel (SL nection). Original -- FE -- PPOFAOA::LLaaccttaatiioonna!laannddPPlaacceennaallTTrraannssppoorrtt PPhhaarrmmaiccookkiinneetticc Stdudyyiin Rants Rss__DuDupboonntt-1133330099 Figure 1 Representative Analytical Calibration Curves 21T2ao7]] YREqeEusaqtuiiaorsnetdfeo=rrtoshsesi3aes5a7dn70ti8ne+ x || Za ps Foe _ || Concenteation, ppm Figure la: ClailbirbartaitoionnscoulruvteiosnshoowfiPnFgOlAinedialfuitteldiwniet)htNoAreNpOlipcuatreepdaekioanriezaedawtaote(rsqaunadremsa)tfroirx matched over a concentration range of0.00051 t0.0.0013 ppm. : i: pe +0 14 REqSuq0au0tai1ro7ne7df5o=r01t0h.e9+9f28e93.3d750t4ine=:X Feo p< | Concentration, ppm Figure 1b: cCaalliibbrraattiioonnscoulrutvieonsshoowfiPnFgOliAnedairluftte(diwniet)htoNAreNpOlipcuatreepdeeaikonairezaedrawtaitoe(rsqaunadremsa)trfiorx matched overa concentration rangeof 0.0050 0 0.0485 ppm. emer ----------5% ----esse-- PPOFOAA::LLaaccatuiioonnasaannddPPlalceanctaelTTrraannssppoorrttPPhhaarrmmaccookkiinneetticc StdudyyninoRtatss Figur2e Representative LC/MS/MS Chromatography Chromatograms DDuupPoonntt1133330099 TSomoooh(in 2x1) 493 " 41 765 WAM 012 cha4n13n5e3l65s0. 7.45] 349 ! 8816v002 592 5267.47hi #4 781 9.06 02 88566 1 5sea Smoomim 2x1) 100 45 07 s02 fs$s77 582 54 00a 727752 802 % -MAM of 2 channelm5is-n 415370 852 s9a5n4125490510002 o S60 eho Tho sho abo 106m0in Figure 2a; iRneptrheessetnutdayt.ivNeeLgaCt/iMveSicohnropmeraftlougorraomocotfanNoAieNOacpiudr(ePFdeOiAo)nirzeetdenwtaitoenrti(mseamispalpNpirgeknitm)teulseyd 49105.2 minutes. 1S0m0oo(tinP2FxS1Oo)A " 532)2 427 48sJ 39 500 ses Smoat(tin 2x1) 100g 136PFOA_ " 1139477.1442 VAM of 2 chan4n1e3l5s 3650. 700884002 735 503820530 933 962 MRMof 2 channels m5i.n 1948125054307004 o Sho eho 760 abo sho 100m6in Figure 2b: iRneptrheessetnutdaytiwviethLCth/MiSntcehmralomsattanodgarrda.m oNfeNgaAtNiOvepuiroen/dPeFiOoAnirzeetdenwtaitoenri(mseampislaepdpirlouxeintm)atuesleyd 49105.2 minutes reser--------r5ereressen PPFFOOAA::LaLaccttaaitioonnsa!l aannddPlPlaacceennttaallTTrraannssppoorrtPPhhaarrmmaaccookkiinneettic SSttuuddyyiinnRRaattss _ Figure 2 (continued) Representative LC/MS/MS Chromatography Chromatograms DDuubPoon1t.313330099 Smootn(n 2xP1F)OA 100 s51.206s Wo] s1e:4r8 wes || 1s0a3e4 as Te Smooin(1n32cxP1F)OA_ jo%s: 112575.662.75163 WRNof 2 cha41n3n5e3l66s95.326e+002 651 5% 15 752 1281S gag 70000 990 MAMof 2 channelEsmS.i-n 4155370 1.775+004 0 500 500 7.00 500 500 10.0m0in : Figure 2c: RienptreemsaelnsttaatnidvaerdL.C/NeMgSatcihvreoimoatno/gPrFaOmAorfe0t.en0t0iomngt/immLe.icsoanptprorloxsiomlauttieolnyo4f.9PtoR5. OviiNnnthse. 071103A20 100 SP5mF2oO0oAth(_Nn 2x1) # 11362149.443 WARM of2chan4n1e3ls53659163664004 o 071103420 Smooth(tn 2x1) 136PFOA, 100- 1358.92.817 % 19555 MAMof 2 channeElmSisn 4155370 1.974e+004 o 500 5.00 7.00 500 500 10.0m0in Figure 2d: wRietphretsheentinatteirvnealLsCt/aMndSarcdh(r0o.m0a0t3o2gprpamm).ofNPeFgOatAivaenailoytni/cPalFOrAefreerteenncteisonandard (0.00306 ppm) time is approximatel4y.9 to 5.2 minutes. - PPEFOOAN::LLasccwttaitoionnaalaannddPPllaacceennttaallTTrraannssppoorrttPPhhaarvmmasccookkiinneetiiecSStuuddyyiin nRaRtsas___ Figure 2 (continued) Representative LC/MS/MS Chromatography Chromatograms DDuuPPoomn-t1133330099 071103A31 Smooth(Mn 2x1) 100 P5F2O5A * 11352506254 MRMof2cha4n1n3e53l65s0156864004 o re 071103431 Smooth(Mn.2xt) 100 j13a5hPFOA_ * 1138164.1732 MAMof 2 channelmSsi.n 1847155054307040 o 500 6.00 700 8.30 9.00 10.0m0in Figure 2: `RceopnrceesnetnrtaatitoinveoLfC/0.M00S3 pcphrmoomfatPoFgOrAa.mofofr0n.e6gamtigv/emiLondo/sPiFnOgAsowliutthiornetdeinltuitoend tS wgminal time* 52 minutes. The measured concentrationofthe representative solution is 0.613 mg/mL. 071103340 100 SP5mr2ooo7ath_(Mn 2x1) * 141547.97 5 MRMof 2 chan4n1e3l5s3659181004004 o 071103440 Smooth(Mn.2x1) 100 13S6PnFOA_ * 11491603552 MRMof2channelm5si.n 1947125654307040 o 500 6.00 7.00 5.50 9.60 10:0m0in Figure 2f: R0epnroemsiennataltciovnecLenCt/raMtSiocnohrfom0.a0t0o3grpapmmofoflPowFOreAcofvoerrnyedgoatsiivneg isoonlu/tPioFnO(A0.w5i8t4hmrget/emnLti)onditliumteed saoplpurtoixoinmaiste0l.y6250.2mgm/inmuLt.es. The measured concentrationofthe representative recovery PFOA: Lactaional and Placental Transpor Pharmacokinetic Sudy in Rats Duront13309 APPENDIX B NN INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY DURING GESTATION AND LACTATION DATA = PFOA: Lacutions! and Placental Transport Pharmacokinetic Stdy in Rats DuPont13309 INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY DATA DURING GESTATION AND LACTATION EXPLANATORY NOTES Note Day 21 = Gestation Day 21 Day 22 = Lactation Day 0. Day 43 = Lactation Day 21. AN g i z i gd FoRPomioiii g BLL :g iSaHbo : i Bie ggos ~ B 4d 88 gEadoTDI Ill EI dro i i jRERsE saa cE [I] 3 CF - g 3 i ii 248g238z2% ticcioioscesancs piiiEEiifrrregct 2 fb EEE EEEE E RE EE Biocnccancszsas Co 6s 2 : z i 3 ; PPoomEs PoE i Bois gal 0 Tad io 08 Pioiis n ii HARE i Blzzpgpiic 3 [IEEEEEE NE HEREE EEE 32 g= H3 iss ses Gs ~ 3 of 83% 383 3 3 3 PogBiimaanl C Pops PoEDaEn PRE 3d EEE i z HA EET a Ek 5 ggo:0 i{HeedEpEsf d8 aBEgBE :1 RiiIiEmiLgIilEnEg f 8388 830 08 of of 3i c: osHho. CE BZ Nk 8 1 Eoin & Eni gdf 000 1d 4 444d" & LLL ELD IDIIII TT N J Of Es3s1iJ CLD d E004 : & g 8 FEL riiriiriiae if ogre i : pREEEES RES a a er? g y ; : g i Ho 3i Pog . 0 oo : PoE. J 8 omin : i Bebo : FI ioe : iz PEE Zz i 3g 8 : fiirriEiig : - isddzeszice i i gi ;Hisaseres &9 ~| 2 i dss sess sla fog cE gE oC El Pron go0s El00aElo gE oC o0 oar |H gB h o EEE i PEE Fas i E3 iPR8ES 89 as ESeas Doi f12 iigidBeEaEndE HdEeuEsHdE 27 i oF of oF 38 of i ETH z ; diese oes \ 2 1 3 i z i gana Pools Pom a Al EERIE TEN gif 0% 0. &88. 88 8 = g08 REBAF o+coeoe ooeensEBiao.oens oo.o= 38 dbllirddria lr fRE ERE GER EEE EEE Z| 8 i 8 5 x Eiau mg Es EEEgE BEE i gE GEIEETEEEEEEEEECE z3 8EEEEPEEEEECEEEEE EEEEE EGE] EEE = CPRRER IERIE RIRGIiG 3 BOREL LLL. 4 oak ms 8 g HTT RE RAR RRR 77 2 v : 2 Hoo iEg ge a ii c PoomE} ddo PE od sme 1 1 Ho EE] EH 2g i 3 3 i 3 ER RRERE NE B5oe)l c ix g,Eiiizdizfa8isfxidfaizEiaEaEisE 3 Li EiEEREEG i `IEEE EE EEE 3 ig 8 LEAR RAR AE 22 : ~= 2 i Z| al - < 2 oe = si dn Poe En BEL DE Eff CC E P0ogEaEiElo Jf1 E [ oin , i2 qo. oa 2 2 3 g Palpder Rad sf iia Ha :1 27 f8 of goEf aonf E5E3E30aoEf 73 '. 2 i 5 i 5 Bb ii :i al a ] 8 ]a z Hood Edel i gal (dC 14 14 44 4 8 gL lll : goood g gif ood 14d 0000000 ? 3 FoF os es es + +2 Z 8 EE 33 sflllllieriiii io jigeeae reais qo: gE 0 &, z FE 18EEEEEEEEEEE; EERE RRRRRE EivrzsssgEusuuacg fisagggaiaaagagt 2 = (i335 RRERAEIIRLG a ER LLL: recess / 2 : 2 i go ogi oF Poise goglih Eoogal lll Eom ios 38 dono ig 2 g 3E : Pees PPEgs ial8888 gb iEiEfigg 0 2 ipzoaagEEEEE 21 fi BEiEiEfyiiBeEeEEe:: : gs Hie ss zara 7s . >. 8 i goFE B oAi %cai gel 00 Popa Posie Poodles o ~ 3 oz aloo i : jes dodo Tigi os fas feels i i EEE I3 F : og P5 Ii EpRpzREz GE2EE2 EEEx 1: CimiuaaEdn :3 gi E38f:ile or o woe 7% PFOA: Lacutional and Placental Transport Pharmacokinetic Study in Rats DuPont.13309 APPENDIX C NN. INDIVIDUAL BODY WEIGHTS DURING GESTATION AND LACTATION 76 PFOA: Lacttional and Placental Transport Pharmacokinetic Sudy inRats DuPont 13309 INDIVIDUAL BODY WEIGHTS DURING GESTATION AND LACTATION EXPLANATORY NOTES Note Day 21 = Gestation Day 21. Day 22 = Lactation Day 0. Day 43 = Lactation Day 21. Abbreviation = Noda A SE 7 PFOA: Lactation! and Placental Transport Pharmacokinetic Sudy in Rats DuPont.13309 RIDEVISUAL BODY VEIGHTS (grass) DURING GESTATION AD LACTATION Grove 1: 0 vrxaony nsemns ows ms owe onesoowx mas owe ws maw 3s3 aBFitaAo nT aGramltes pdvmnihy aavmeeas EaBeiSs adEsessoer] aewea i53 BBSaasas dMmeaaalees sdaeemdnc sadmeesas dddenaasd mddaiess gedeeesss 355 BBleeSelOHemmaSs eaBmentn maGmYdd dmRmEia dmhaannn aIaed Fi33 fMHeestr dHaens EBeen Bdeeells BpBEiyd dhhimina aAadn @aH BSBalUdSOGdMReISE Gaemenan aadmnedon mmamnse BbBeeOdd) Ibaniens 2ia SMPeEaaLss Bdnnille addeeesns abammaaoo ddbweesga eBwseose ebheuisss 5w fmeesd EEeDl dWeLn dmoRn Bmand dBann gBua iFG2RsOUP MEAN 2W5R 4i.ao7 O2WU 63ie.1 S2E W6i7ao.s8 2WiOmoN.dE2 2iaY77ome.8 284.3 ioBR NgIiaonii 0 PFOA;Locutions snd Placental Trsnspor PharmcokineticStudyinRats ________ DuPort:13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUZ 1: 0 Wa/KG/DAY Tannax DAL DAr12 DAY 13 owsDAoYn14Test DAY1S Dav1s Dar 17 25 15 23220 20s 20306a01 s2mo3rs8s B6delldes 3awi0as a38m:7a3 d3e5z s0 2Fi0d a250007001 armsl 2817 ammlrs 284 aalo 293s aaee 2095 :: : p:s 35 S2= 0 ml. s 0- ab- e is>is -: Z: 336 3 306=.5 22.8 05- 3 2955 aw- a 30.6 am- a 3086 aan-ss Eze-ad sais2 ae 35 3 36:.2 - w:s - ad:- a 23a : m02= 2 Me`= s de:es = 8& 320906..47 229997.55 3006860 3a3s7a 3nlas waliee adadse i& 32152313 32923s3 m30e2ss 3nl7s admeea d3amz a3n0s0 1s z1ms zea sea jon aes ans Raosve wews i29d8.e6 d3a0d1s2 130270.32 3160 mDiags aaa xXSma asbe eve sk iases i3 s i3s28 ias2 iasse iaore isas 7 -- PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont 13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP I: 0 MG/KG/DAY an=o= DAVIS DAY 1S DAY 20 DAYSDAYon21TEST DAY 22 DAY 2) DAY 24 125 a3s0i.n1 asesdn assles sasoora :: :: -z F1d :- :- :~ -: .: .: :: n20 :: :: -: :: .. z. :: 23 -: -z z: :: .: .: :: 331 38 si> s se: s wi: n an- a -: 39.0 387 ws sa - :: - :: - 5a siz s 33: 0 a:s 39: 6 ph: :. z: &52 e 36:.3 3907 30:.6 mele 1:2 39703 ao2 s a0 28:a6 ams 297-.0 desis 293:.1 332 7&s 8 336670027 3slo a3ee1lls7 slo a3se1ls amin aailsalse amos 2a3m5e0 ama 3m0r00s0 ames 3e1ds ais > sGroo.ve mew 136.2876 2m1e4.3 239153.58 a1w9..02 ea2s5a8 214m43 wRoiss Nse 6i1o5 ei)s i6o7s io2a 21355 el5 7.6s6 EJ PFOA: Lactationsl and Placental Transport Pharmacokinetic Study in Rats DuPont13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP I: 0 MG/KG/DAY NanEnR DAY 25 DAY 26 DAY 27 oarsDAYon28TEST DAY 29 DAY 30 DAY 31 15 :. z. B:s :: .: .H :: ois 20 -: : :z -: :-- -:: :-- z:: n2y -- :- :- .: :- :- :z 332 :: :: z: .: :: :: :: 33 :: :z -z :: .. :: :: 5a :- :: z- .: .- :- :z a55 & 307-.4 31 307-.4 3000 al- s lz 30-s. sais 323-.4 pels 30-.6 els 335=.0 sie 7i] adllis 33sm aselles 3s3l0s 1 ae esliss dsaalee a 290701 2ssl1 30ls 303s 32005 dma asa sroove xeaw Sie! 31153379 sss 1387005 750 1395.478 el 138:.2 eas 1a2.008 ssa lauage Ne1gNed eal 5.30 s 5 5 5 5 5 5 a "PFOA: Lactationsl and Placental Transport Pharmacokinetic Study inRats DuPont:13309 TDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP 1: 0 Ma/KG/DAY anMnBEaR DAY 32 DAY 33 DAY 34 DAYDSAYon3STest DAY 36 DAY 37 DAY 38 i2s :: :: :: .: .: i Bs 2n0 :: :: :: z. c. :z :: 32 2 :: : :: -- -: :: : --- :: 336 :: :: :: :- .- :- :- 3a8 :: :: :z Bs. :. :: :: 555 a -: 387 -Z mle -wes -mile -: wa -jes -3200 &a 75 33606121 FC 3s6e7l2 ICT 3363972 HOES 3d6e20 HE BE 33586100 1 SE 3d6s3e7d YEO ddeesiss +564 13 32sle awla 3aslz amis sale aes sie G5R0OUP MEAN 3d63s0 14873.62 1354.65.05 1370.2898 3164.2071 3485 xNg.0 154s 102 se 6353 se5z a15 sss ee5r 7050 alsr 0 PPFFOOAA:LLaacctuatoinosnll andPPllaacceenntalTTrraannssport Pharmacokinetic StudyninaRtatss DDuuPpoonnet-1133330099 Gov 2 soman 255 0i5 335 3%38 2a5 a"a 3i SDIVIDUNL BODY WEIGHTS (orans) DURING GESTATION AND LACTATION 0 worko/oaY oa 3s oar ao oneEs aow zest oar a2 oar as :: -:f :: : B:: ::: :E: ::: ::: : ::: : : i i ::: ::: :: : : ::: ::: : ::: sBseeon oi mwne wRanand mNMiEEaE aaae) ssesds mmens oawhedd BBend haid G$i55REOUP MEAN 3hT36ae.3e0 f33We5R.:w5 3n3m4dE.$2 3B3h1h.E0 3E2N3RE.:0 N" -------- w e-- eee---- ------e --r PFOA: Lactationsl and Placental Transport Pharmacokinetic Study in Rats DuPont13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP II: 3 MG/KG/DAY WaEnRn DAY DAYS DAY 6 DAYSDoAwY7Test DAYS DAYS DAY 10 23 2206860.38 2ze1sl1z 22776..08 226.3.04 2e8s9.s4 221024.79 2a9i6.e5 220 2a0l30e1 220e7a.l6s 223563l60 226631s0 224679.036 a2e1s9 szeaade 23s0 aass2lio1 a2slelss 227660l000 226s6e8s 229606090 2279448 237165..81 a aassellsa 2a3e1ss a2s5s0i0ss 23505s2s 32065087 a268e01 3270s05 Ians 2233920 2e6e0l7s aimais m27e5.s0 2am7e6.4 zseerea 2aa9n85 s5t0 226790s3 2we6l17 2m6177 2a7s470 3a7w6.a3 saesbida z3ee4 5H 22063050 226690l7s aramliess a20n.e1s a2873e.l10 228e9s..88 229878.000 8a 226a2s0lss 2as7s4i7s 228612.s4 22m6e6lss 22910803 321091008 320887..27 E7"S 228025 2a139l7s 221e7s.l7s as2ls02 2as9e513 239961l3s 230963.40 776 22545077 2ass1is 22aeislls0 s2ye6sl0 2a6m0yi 2a65n03 2251597 . Rsoo.ve wemw 215375.s3 21602s5 21690.6 2174s.9 2l7e8s.z5 21896.14 XNege.e0 sz 302005 32304 220% 3200s 33200 2a02 a30e w= PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP 11: 3 MG/KG/DAY WanzoEnn DAY Il DAY 12 DAY 13 oatsDAYon14Test DAY 1S DAY 16 DAY 17 s2 3208002s 3as0s7i7s 2aew9les 232ms 237a. wanise 33576 4 220 22087073 a3s0o0l0zs amslsiss 3o6u3s ndeasies o3e0s 3d2e1 e 3205 20-.1 298-.4 299-.6 33-.4 3262.0 =: -: a0 32s6ia3 32s9e0l7s 3200s1 2352ela 2369s93 32: a : 0 aa 30- . 30-77 30- 2 me- a 20a. :: :- 55506 S2929:602s 2m99:s1s a30ws: a smso a 3aa:a zz: z:: 5ss :- :- :- .- .- :: :- aas 2299527 s2o3l9s6 332s3 m3lms 0s8u.i4s 30 0.8 4am7i8 27 23876037 a20l0e1 m30i0llos d3e0lslso B307L.6 e319s01 i33s43 . 7 2203 2206 212.1 2m0ls 2934 30s 31s GRovP WEAN 297.2 300.4 307.8 31ss 319.9 3214 <SNQLO sSoE i5.5e 2503505 1a9.31 wa e s2s 04s 132.57 148.8h9 is is is is is io io 5 PFOA: Lactational and Placental Transport PharmacokineticStudy in Rats DuPon-13309 THDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROVE 11: 3 MG/XG/DAY MaonnpoEnn oAY1s DAY 1S DAY 20 DatsDAYon21TEST DAY 22 DAY 2) DAY 24 2 3327.061 slo7.4 aaasne asemlas -- :. -z 22 2s a3s3lse a3e0s0 3deasis sa0se7 :: : : : : ps z: B :z: h3a0y 33:.3 si- a se-ss 36-50 : : : : : : :: : :.. ias 50 :: : ::: :: :zE ::: ::- ::: 55 ::: :: : :: B :z : ::H :z : :: z ase 3.- 0 35-.3 aa aSs 295:.4 20:6s 35:.5 n& 33669l0ss seellss aas0ssz asmeas aass3 I301n a30l8.a6 2E%S 335001 deelsiss 3I91n4 3w98a7 zmesslas 2m9i7ss 3s0s6a.:3 GSroo.up weaN m34ss. e m36a3.e5 m32a.n6 a39i6s5 e32n8s3 12939433 x 166s0 sN z islses 1s0ee 17.003 7t3y2 28558 5958 7s5 % PPFFOOAA::LLeaccttaotinoanallaannddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSttuuddyyninaRtatss DuDPuPoonntt--1133330099 GROUP II: aNnamneoR INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION 3 MO/XG/DAY DN ZS DNC 26 DAY 27 DAYDSAYon23TEST DAY 29 DAY 30 DAY 31 22 25 :: -: :: :: :: ". : - : : : : . : 300 a :: :: : zz ph: z : z: - .. :z z aas 50 :: :: : zz z :: : :z : ... :: : sste 5 :: : :: - :: : :: : -: : .: : ::: 5a8 & m-e 300s m:o 300 3:s 3087 23357.00 38:6d 305.8 MlS s 291s e:s jos 721 330091s2 20700 3s06l13 s 3013 M3e1a3 3213 diellss 3017 3m3a1l0s 305s sdaedse 283.0 s3iad 3064 GsRoOUP MEAN Sle 2L0e9d.e7 64s 3L1r0 slo 131690.16 72 7200626 90s 2356..433 1121 232503081 lool N2e1.8s sler N 5 s 5 s 5 s s 5 PPEEOOAA:: LLaocctuuttiioonnaallasnnddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSttuuddyyiinnRaRtasts___ DDuupPontt113309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION Grove 11: 3 wosKa/oRY fr ER oAY 32 DAY 33 DAY 34 oavDsAoYn3r5est DAY 36 OA 37 Dar 3s i22s 3 ::: :: :: ps :: : : :: : : : : : : iai : ::: : : : ::: ppss: p:s i55s : : : ::: ::: :B:s 555 : :: : ::: ::: : - 752 B0soi5ss sameuilloa 3s3e23s 3dweses asassia seeeesss d3osve 77 sBeel mWeEl m3l3s 3d0a3e Maea eaeEe mana GsSiooe wen H Bpos3:a1 3 amweessa 5 swns.iee s 2meedanrl : ew10dss s aa1nr%e a2d3se0 5 5 rec ----5%e ------s --Sts ---- POPFAOA:: LaLaccttaatiioonnaallaannddPPllaacceennttaallTTrraannssppoorrtt PPhhaarrmmaaccookkiinneettiiccSStuuddyyiinnRRaattss ___ DuDuPPoonntt-1133330099 INDIVIDUAL BODY WEIGHTS (grans) DURING GESTATION AND LACTATION GROUP II: 3 MG/KG/DAY pNrgER DAY 33 DAY 40 DAYSDAYox41TEST DAY 42 OAV 43 2i -z .: -: :. -z 222s ::- --: z-: :-. :-- 300 : :: :z -- :: aa :: :: :z E: :: aso :: p:h :: .: :: 5s1e - :: :z .: : s5s -- :: :- .G :: 55 = 2398023 330s 2322032 33 2s9e9e0s1 3eslo s297i04 i asin a30s0a2 das 77 322071 2sess aBl1s7 3m03l7 d08s0s8 . Rsoo.ve MEAN se 2362.78.10 Bir 2372.7.380 12021 3293..062 12lss 32266..362 10m 3202.519 939 Ne x 5 5 s 5 5 -_-- 5 PFOA: Lactuions and Placenta! Transport Pharmacokinetic Study in Rats DuPon.13309 DIVES 300 WEIGHTS (grass) DURING GESTATION ND LACTATION Grove zzz: 10 wakarone sro nn wre mas owe onsmaenlses ows ows aw i$s: i sHfoeEan fas adMmensn den 2md6en0n ah aDBmieane bas aRfeeddd Ban aBhmaNan ded sBaer es iiit Fo BrmoRe Gmmn Beloofnl dmdosmn ddaeeEn Bb+ieds sR ea s a+ad Em ne 553 5 RhaEesd Ses ModmeeeesSn dw amImSns amis mBmaEsn amo dRIUmnDi Dwe mmBilna dgn a mheensl i@= a BmSe0el0 Hes GdFamelsna Gs gmaeamdnl et BBdeaeodd dma BRseiaadds ama Giegss ama Ie anene &@5 = SmEeeOe BRdeEn o mes aml m0an dKmeRdn md ne hes EEdnSs REDO aromiers m3 . G5oiRkOUP MEAN 2W5s2ia.oo5ei2i6sn2o.o4 2R6S6FW.S6 2mi7N0owm.0 277.3 NiEo 2B 8B4i.E4WNaWNsiyo - Ts PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont. 13309 GROUP III: NaunuoEnR 5 pr1t 1i6e 211 258 335s aie 0a a&s 7" G5.o0.ve wew S5 E INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION 10 MG/KG/DAY DAY 11 DAF 12 DAY 13 DAYDSAYon14Test DAY 1S 23.1 299.5 300.0 304s aise 320620.30 3290780.12 3303ss mjose.s 3n1as z 242 - me-- s we- a > l-a - 360> > 22002092 22770s2 - 22m0.0s0 - 229720.53 - 23903.26 . 33007:.s0 ai- a 309.0 aaT a aie me- s ms 3355: a0 3a0s60i1 203 3a0m6ilss 2708 3Wi3s 3091 3w26ss 3090 a32salo 3187 2206110s6 amis 2278al7s sls 220:0021 als 22093820 slr 330005.03 39l3 sols 31803 3asls als as 2I9.s2 30 aimeds 42s 3ims.o aos e16s.8 a3 sodao 500 is is is is is AND LACTATION DAY 16 DAY 17 - - 3-s 33-43 hs- >: 308-.0 315=.3 331-a6s 3362s9-01 ps- -: 36-.0 376-. 3ma0l1l0s 33340s8 aslollos 3d3e:e3s3 34 desis 2W1L.I1 \Visgn e0en 6.i9o0 oF PFOA: Lactation! and Pacers Tanspon Pramscokinetc Study n Ras Duron 13309 IBEVIOUNL BODY VEIGHTS (ans) OURGNG GESTATION AND LACTATLGY ano 131: 10 worKG/DAY WJhe ow owas own oatsaoan rest ow: men nwa i Wo sr wea wie C : : 3i wi?s we:a ai'a a:a C: :: :: :ifGltsove mem W3t0t ae5 w3wesm w BmeuEzm Wawseno ou wue3a om30s:owgowne! = PFOA: Lactation! and Placental Transport Phasmacokinii Stuy in Rats DuPont13309 INDIVIOUAL SODY WEIGHTS (grams) DURKNG GESTATION ARD LACTATION croup 132: 10 wo/KG/oAY smeos savas owas maar sasmaornaerast ow oarao oar a G$ii :: ::: :: ::: i ::: ::B iii ::: ::: ::: : ::: : iii ::: : : ::: :: ::: :2: i3a ::I ::: : : ::: : :b: aai a meie: ad mdBimn ia amiHno saem:eo mwei:se awie: sl Baee:ds Baro od dan Bn Bnd Rd ml 5n% e Wnon aamndn dEni 2 3S mEdr uess GeirsEove wan 2aR2aa mam0lEe mamawEs mamehEs nRowsRe BaBilRa wNmpSne :::::3 5 PFOA: LacuandwPlaicenotalnTrasnsport PharmscokStiundyeitnRiatcs Drone 13209 INDIVIDUAL SOY WEIGHTS (geass) DURING GESTATION AND LACTATION Grove 112: 10 wararone sSoenne oes mos owas onsaaonnmes mw ow owas iii$ i ::: : ::2 ::: : ::: : :: :::: :2 333 3 :::: :: ::: ::: ::: ::: : ::: 53 : :: : : E: : :2 Hi@ 5 sBe:ilo EUs wdee: re Bed adsmwhees eaa:en awd edailea nei:ss We aaee: ds 5a BdEeS sdWeel oowasn daemne mBsh mmeS ad . G5oHReEOUP MEAN wE33iR3.:nC3 3o4o5w.h8e : 3ate4t6.J:e9 3a40.h6 3 3M81h.6 3 R351N.0 3 NaE4 gse: 8 -_ 5 PFOA: LactaatnidPolanceantlalTransportPharmacokinetic Study in Rats DuPont.13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP III: 10 MG/KG/DAY ameoRn DAY 35 DAY 40 DAYDSAYon41Tes|t DAY 42 DAY 43 Hs :- -- -: .. -: 113 -: -: H- -. : i1 -- ph: :: :- :z i2s1 :- :: z: zg z: 323 :: :: :: .. :z 33s :s :: :: s. :: aas E- :: ps: .- :: &5 os 38- 7 2s 30- s ls 35>79 ded Ess i : tsoens in5a 3alds 3807 3306sla2 3eeln 33s0s8i3z alr a20e91ss 3s 23935060 3m GsrDo.ve MEAN sz 32309.03 slr 323081 mlz a28241 Lim 3B4.s3 lelse 232665.08 iiae N\.N N 5 5 5 s 5 _-- E PFOA: Lactational and Placental Transport Pharmacokinetic Study inRats DuPont-13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP TV: 30 MG/KG/DAY amBoEnR DA` 4 DACS DAYS barsDoAnYTTest DAYS DAYS DAY 10 31 H 2a6i3l.s4 2s6ls 2a6i5s.l6e 262ls 2a0m0.l0 2681s o2a2si0s 2eals 2ze7sima 271s 239.9.80 219.3 2z9ee8.0 275.8 110 26 22699023 23401 aasmlios 2u3ls m26eslas sls 2zamls 2s0ls a7m3ls aslo e23a0 255.0 221s3038 259.8 225 220881133 25308 22s3ilsa 25513 228197030 2600 ezeasis 2650s aaese. s aesld 222116.9081 32s7ea0vs.ss5 2i as a26i8l0s2 mls 2aa0y7 alo 22assllz2 als 2ae0s ame 3z5e33 zee 336925.36 80 220e6s.e4 295% 552 Ha s2a7l0e1 sls 226600lss 230 2265611.36 2550 226762.4 ze 227653..48 srs 231856.s4 sel 2Z9ee0.8 297.4 7a70 2250601ds 25006 22sse6llss 25001 eesis2226 57s 28905 assis 22a7s4l.s2 26s. 228776.00 seen s28e0e.d4 211.8 77 2203101s3 2s6i0l2a 22666.34 2ze7e.l8s 224s7 2s7e6rse asraesss G5.roup ea sie 2105.6010 220 s11e.9d5 268 6 1e.08 3s 2106.00 255 a12m.4a3 28 21173s4 RNN10e8os 2.51 2.36 N 20 20 20 20 30 20 30 ----e-- e-- e--------%---------------- PFOA: Lacttional and Placental Transport Pharmacokinetic Study in Rats DuPont. 13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROVE IV: 30 MG/KG/DAY nMBoEnR DAY 11 DNf12 DAY 13 DAYSDAYox1T4EST DAY 1S DAY 16 DAY 17 31 5 97!.8 3000.8 305:.0 37!.4 392.2 Me`ad ne s 25.0 21 2910 2395 310 : - 0ry 26 :3s 26~- 7 :: 2a -2 ase -293.7 :: E: - 22 3 22m07ls0 als 22007032 22203 a3s0si3s 2sel0 2os9l7o8 does 2a9tsels6 dese ms: s 3a ne= s 3s 28 as a07llss 30612 aemlly 30sla ademliss 309i a33e0s0 317s a36s0 s 31s 3s:- -: 33.7 552 5 il- s : 30-02 - dw- s - 31-63 - 3-a - 3-8 - da-se - 7a n j2oe1l02 aris 229e.sl0z amis m23a0s3 2911 23o2sl0s 2082 229076.38 zea 2306072 does 33067..24 3056 77 20301ls 22ss3all0z 22sse2llol 330mslz 3J0esns 337s damavse Rsoo.v Mewv se 218039.51 28 2T9i1d.e4 zs 21927..65 326 310267.55 328 31028..638 31 3la6do \VRege0.s2 ads 44s is is is is is io io -_-- 57 PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont-13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP IV: 30 MG/KG/DAY NAanszear DAY 18 DAY 1s DAY 20 DatsDAYon21Test DAY 22 DAY 2 DAY 26 31 i 302.9 - 3Sa : 395:.0 E a:2 : :. -: :- n] 26 :: 2 ZZ - -- :- -: : -- :: : 22 3.>1 36-.6 as: a a-e :: :- -: 23 IH 360-.8 - 36::s. 39i:-.8 ai:- s :-: -:: ::. a2s Ed 336s7.05 3eems aawsls a39k30 z- - - . - - :- : -: a6 7 37-.4 adele 33- 3 msl 39290 3290 a-a ms oas-is 1a 20253 ass 31- a 2900 77 7 330201s3 sels 335890ss 3632 33000 63 sasso se a30m7.e4 sas d0 ees 2306 22939100 288.7 R5o.ve wma 31836.60 3as2..d22 I20T.8S1 336L.s00 2031.319 263.s4 NmeSgets 5se siaos islosz i1o8 loioos lol5a 2.955 ad5e % PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont 13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP TV: 30 MG/KG/DAY pemes Ar 25 DAY 2S DAY 27 DatsDAYow28Test DAY 29 DAY 30 DAY 31 13 :. -: -z -: .: -: -z 105 i -: : :: : zz z :: - z: : :.. :: 22269 :: :: :z :: -: :. B:s - - - - - - - 32 a :- :- :: :- >- .- : -: a52 5 ::- -- -- -- :- -- :- a5 7 am- a 300ls aw- a 293la aw- a 3osir Bi- s 30513 30- 0 3091 w-e 2eeld de- e 2s8s 17 id 3a0l:s3 ela 230957103s 303ls o3aill0 300s 3s0i4n7 30305 3a0m6as 20913 2msse7 jes. 330a3.8 3022 --_---- G5R0O.UP MEAN siz 30B06.61 ae 30s3e.z8 ald 31120..965 sls 31123..00 sso 3La4d0 sles 3n1d0.s4 sa N1GdOr ae H s 5 5 s s s s % PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPon-13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROUP Tv: 30 MG/KG/DAY aNnBER DAY 32 DAY 33 DAY 34 DAYSDAoYv33TEST DAY 36 DAY 37 DAY 38 31 .- :- z- :- - :" -: 0H -- :: :: :: B:s .- :: 215 :: -: :: :: 5: :. :: Ei2 57 :- ::- -: : z-: :-: --: : :- 82 i -- :: -- -- :: : :: :: 5525 6 -: - -: = -- -- :: = =: - -- 7a0 n 336600 3s 3333sa mie 3313s9 307 336ss a a36m6sd ale d35e8e8 Jess d3o1i2s 3050 77 a 363 0Bi3s mBUle male mussed mdiess s33ad . 5R.o0.ve Mews 2322.2471 322173.s5 312s3..5a 132362.03 2310..898 23.00.88 xS2w8e3d0 ~se 98s5 sless eas sz5 s5e sa5z 1a.e56 00 PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont. 13309 INDIVIDUAL BODY WEIGHTS (grams) DURING GESTATION AND LACTATION GROVE 1: 30 MG/KG/DAY aNnunMBER DAY 3S DAV 40 DAYSDAoYx4T1EST. DAY 42 DAY 43 31 : .. : -: : :. . -: : -: : 6110 :: :: ZD :: :- h ph : : : i29 3 h:s : -:: z5- . :- : -: 2i i ::: -: :-- --. --: 5E2d :: :- B2s .. :: 7Ia 38:.4 2061s 355:.3 2090 355:.2 302is an2 s 29els as- o doit 77 3S27s2 omllss j3o0es saors2 assasias 7 alt 3s 3esla 30a 303s G5.r0o.up weaw se 3223168 les 32232.82 ll 32.23055 10000 3b4a9s rez 219.202 esi w s 5 s s 5 . N\.. Tor PPFFOOA:LLaaccttations!andPPllaaccentalTransport Pharmacokinetic SStutdydinyRaisnRats_____DDuuPpoonnet113309 APPENDIX D N. INDIVIDUAL BODY WEIGHT CHANGES DURING GESTATION AND LACTATION -- -- 0 -- -- PFOA: Lactrional and Placental Transport Pharmacokinetic Study in Rats DuPont 13309 INDIVIDUAL BODY WEIGHT CHANGES DURING GESTATION AND LACTATION EXPLANATORY NOTES Note Day 21 = GestationDay21. Day 22 = Lactation Day 0. Day 43 = Lactation Day 21. Abbreviation - = Nodua A - PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont13309 INDIVIDUAL BODY WEIGHT CHANGES (grams) DURING GESTATION AND LACTATION GROUP X: 0 MG/KG/DAY aNmEaRn 410 0-15 asa oars2o2n-28Test 25-29 2-36 36-3 1275 2w6o3 3-0 an- e :- 285 0s ss - -: -- :: 12750 30000 s3i1s -3s alr : -:- :: : -- :: : 23231 328s0 2a800s 3Z 3s - : :: - :: - -- z- 336 36 03071 s-ea 05- a 31 me ss --: :-- --- -- 053 55 s1al2 s-o r-ea -: 30 - - - -: - :- -: - aa & 2a20 a3 a2msa wie eesz amsisa sls els 17.20 sls 0 187 isel as 2005 7a 3a638 2mellss waemlllz 1si0s 1w0l0 2lss mal1s Gsroo.ve wemx 3B4s5 373d8e m9032 se0s4s 1e3a 1s2a0s x"Ngo dae xSE 120s iiisss aiios 20250 27sa 3 5 da5n --- Te PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont.13309 TDIVIDUAL BODY WEIGHT CHANGES (grams) DURING GESTATION AND LACTATION GROVE 1: 0 MG/KG/DAY NaoMmBEaR ao2ars on 2TESaT 57 176:.1 -- b1o2 11- 6 :: 2uo0 :- :: 33 :: :Z 332 10-.4 z: 538 a ue2- s z:: 3553 16- s = 5a6 1m6i8s a2las 7as 1was 1a6l09 0 Bz az ? sRoo.ve wer 152.96 53.20.08 N\ wsie iaoss al5m 105 -- PFOA: Lactatonal and Placental Transport Pharmacokinetic Study in Rats DuPont 13309 INDIVIDUAL BODY WEIGHT CHANGES (grans) DURING GESTATION AND LACTATION GROUP II: 3 MG/KG/DAY an ER 410 10-15 as21 DAYS2o2-n25TEST 2529 29-36 36-43 22i 223s0 3a0l7s saislss -: ui sls owls : -: : .: : .5 22 30 s50i1s 247 30-03 sw. so-ls - h:z <-- :-: ::: 0a 3 2 32 302 23as > s-so : -: -: : -:: -: E a5s0 si 332 3323s -: pr Sit] - :: : :: : :: : Zz : 55 5 250411 :: 07 - -- -:- --- :- :: = 5& 2 32425 28s 22d0l0s n3l8s3 msrls 2501 er losis malao 30s a3n3 a3nh 3s sls %7% allos awslao wseil amslisn a3l3s 21030s nilsse G5.r0o.ve wey 1302507 SE 236 352s1 2806331 1598.238 131.63s6 an7s7 \N8270 238 642 26.05 66 5.05 3.81 20 is io s s s s eect106------------------e PPFFOOA::LLaactatiioonnaslaannddPPllaacceennttaallTTrraannssport PharmacokineticSSttudyiinRRats DDuuPont.13309 INDIVIDUAL BODY WEIGHT CHANGES (grams) DURING GESTATION AND LACTATION GROUP II: 3 MG/KG/DAY aSnngEs ao2ars on T2E2STa 2i 11562.2 -: 2220 11762217 :z 320 :- -- a0 13:.5 :- pra :: :z 5a0 ph: -: 55 :: :- 5& & w-a e-s ss ssn 27 Ea iwlls saesds ws ols -- GsRDo.uP MEAN 124046.64 507..809 No nsN e i75o80 26.7s8 -- eee--10-- 7 re E r-------- PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont13309 INDIVIDUAL BODY WEIGHT CHANGES (grams) DURING GESTATION AND LACTATION GROUP III: 10 MO/KG/DAY amMaBEnR 4-20 10a13 as21 DAYS2o2w2sTest 25-29 29-36 36-43 3s5 4i6.l0a a=s -: 37s 32 : :: 3z : .. : -: . 116 2al3s6 2-8s e:nt z: -: :: :: 11 w57s.0 a-o Bhh z: :. :: -z 22 2$2536 22.0 eseas E: :: z: .. 33 39 2327s3 0:a am: a :E 2801 - : Bs .:. ::: z.z ias 3328s ailsss -- :: :: :- :: &a 6 2836 u31s eseasb 96- 9 0 ws els ies :53 2-08 23 :5.0 als I7 7 220065 32 0341 wl 7e8sio soo 11ss 16s lwood &5 210s5-19a0l5s 2s 24.8 G5r.ove usar l3d6.s4 ss 235 3S5B3 msiedss oasssa si ase 2128 2105.08 130 1s8da Sa1R9sS a0 74s 30 is io 5 5 5 5 - a. PFOA: Lactational snd Placental Transport Pharmacokinetic Study in Rats DuPont-13309 INDIVIDUAL BODY WEIGHT CHANGES (grams) DURING GESTATION AND LACTATION GROUP III: 10 MG/KG/DAY anBEaR aDaAYS on T2E2S4T3 35 -: -: 11 57-.1 -: 1168 -: -: 121 i:s p:s 25 m:i :: 335 195-.8 :: ias & mi:- s --: aos umsi a07's 78 3 13 11022 7 ms 2 . R5.o0v.e Mew 12509.67 aal0k.s7 se 637 15.59 ANN w io 5 --ee--------------------0----------sr-------- PPFFOOAA::LLasccuttaitolnoanla asnnddPPllaaccenetsaTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiicc SStutdyuindRatys inRats___DDuPoonntt-1133330099 INDIVIDUAL BODY WEIGHT CHANGES (grans) DURING GESTATION AND LACFATION GROUP TV: 30 Ha/KG/DAY jeaamviaed 10 tas asa DAYSaoaunTest amas 2938 seas 13 : w36o22 3206 sis s:so : - - ::. Bs 2611 312005s7 s::ia : ::: :B: ::D ::Bs @52 3 EB3C2 31 3k um2so : : : : : ::: iaas 2 2i4o 0 a00s0 m2-a : B:: BB:s 2E3S 3 S3005is S::es as>2 s B F:: --: :B:s n7a 7 3nssoomamales md1sls ames2s a3 ma ais ma bee es n 3las mGaillsa mh na Sas . 7 6 Bo awe as wi nh hl GiSEo.vern 2Mis5n 20 am2is0s memolons aGauooss is io g eSmeeoss as1amas StSoaasn : : _-- Tio PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont-13309 INDIVIDUAL BODY WEIGHT CHANGES (grams) DURING GESTATION AND LACTATION GROUP 1: 30 MG/KG/DAY aMnBaER aDaAYS ox T2ES3T 13 10.8 :. 1i :: z: 126 B:s :: 223 3 116675-..73 :-- 2i .- :- P52y 1m20n.2 :- F%S :- :- 3a w79e2 3s8i0a0 771 ssazs 2ais3 7 mz ao 5o0ve we 13381.1 a1s1.a2 N.. sN e Tolieos 13.750 Ti PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont-13309 APPE3NDIX E N" INDIVIDUAL REPRODUCTIVE DATA mn PFOA: Lactionsl and Placental Transport Pharmacokinetic Study in Rats Individual Reproductive Data Explanatory Notes Notes Nidations = Implantations Five dams per group were sacrificed on day 10, 15, or 21G. Abbreviations SE. = Standard Error N== Numberin Group NoDua Dupont13309 A Tm -- & : g i i | aanangzNazsnaen ) . 5 ii 5g | gamgnsznazsanen ]i E1gH ~oUNSRRANILRGAR "ws iN . : & i | anaaaansRsITIRINT . boa : |. RaanIIIANT g Bosses i :z H $i: i | RR3uIERAISE Je ft |EEEREEERLEEETEE - " NN. : igc8| PEEREEEEEEFEERS bi df z ; 83 Grin 2: $A : 28 NeQRSRAIRRNATSS 3 | sgsIsanInnanats 2 i } ~ 5 i ! | [EeIsaaInganets [l 3 Ce 777 PFOA: Lactationsl and Placental Transport Pharmacokinetic Study in Rats DuPont-13309 ~~ APPENDIX F NUMBER OF PUPS DURING LACTATION is PPFFOOAA::LLaaccttaaitioonnaallaannddPPllaacceennttaallTTrraannssppoorrttPPhhaarrmmaaccookkiinneettiiccSSttuuddyyiinnRRoattss ____ Number of Pups During Lactation Explanatory Notes DuDPuPoonntt--1133330099 Note One pup per litter was sacrificed on days 3, 7, and 14 for blood collection. "Two pups per liter (litter size permitting) were sacrificed on day 21 for blood collection. Abbreviations SD. = Standard Error SE. = Standard Error N= Number of Litters in Group Nh - i] PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats ANIMAL a 7&5 13 s5eDan se ANIL 8 && 77 % H5e0an NSE ANIGL 8 66 7& 7 sseoab.n NSE ANDGL# 7a0 77n H5oa0n wsu e IMBER OF PUPS DURING LACTATION Rove: 1 DOSE: 0 MG/KG/DAY BORN AsoLryE DAY DAY7 DAY 14 Dav 21 13 fo1 1 1 11 be1] bt1] 1 bd Fi] 10 1 126 11s2 1216 nis 1140 25 w2sm 12 12sm 12a 128m 12a 12.m La 1i8m La 1G.l6n La 5 : s 5 5 s GROUP: XT DOSE: 3 Mo/KG/DAY BORN ABoLrnE DAY3 DAY? DAY 14 Davai 1s 15s f1l fe]3 12 11 116 13 i3 11 11 110 no n H] n FS 10 waes wa2s p ae21alas a5.e8s d5.i6e 526 523 2: 3 5ss 5ise 511 GROUP: III DOSE: 10 MG/KG/DAY BORN AsoLryE DAYS DAY DAY 18 Dav 2i 112 1o2 be1]2 11 110 105 21 wu 12 1 11 1 n1 1120 1 1 15 n B120 oles Bloeo oles 1r30o oles mLooo oes 1L0oo 04s 110.000 04s s 5 s s s s Rov: Tv pose: 30 MG/KG/DAY BORN AsoLryE DAY3 DAY? DAY 1s Dav 21 13 133 53 12 nu1 10BY 15 2 1H 5 15 be 105 1 5a n 53 10 a9.l2es 20 a5.l2es 200 a5.e2s 20 a8.l2es zor 7a.e2s zon 73380 14s 5 5 5 5 5 H DuPont13309 NNo a No pups. [) PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats Dupont: 13309 APPENDIXG AN INDIVIDUAL LITTER CLINICAL OBSERVATIONS 21 PROA: Lacuna snd Placental Tsnspor Pharmacokinetic Sy in Rats DGEVIOOM LIFTER CLINICAL OBSERVATIONS amour: x poss: arsoroae mVoa senPsYor isihone owsmuanions 6 mo amouncTIES opmacre 66 0 smomazcrzss opened 6 wo smommcriss opmacre> 75 to asmonuaLznIes oanserED 80 to asmonsaLieuas paneer oSsm seEmEen [lEisifrones -- oassarions 61 wo smoumsznies ossscren @ mwa tori smu moss sor Leones sows: 1 sus moss oor 7 wo amomaLinies opmscr> 74 NO ABNORMALITIES DETECTED 76 NO ABNORMALITIES DETECTED waowms scBiEon J wistthaer onsenuncions---- 64 vo assonunLInies oenecro 65 vo AmonLInies ommecro 68 vo AsonunLIniEs omnecro 7 to AsonnLTniEs ommecrED 15 to AsomnLinigs ommecrED oawtek emPIer onacosimi:oaxev nose: 30 wexaronx onsmuntions 61 wo amomatnies opmacre> 70 to AsmonaLIeies oEcscrED 71 vo asmonaLierss oprecreD 7 so AsonuALIeigs oprecro 78 vo AssomuaLimiss opnecreo EE 123 DuPone13309 AN . -- PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont-13309 APPENDIX H A MEAN PUP WEIGHTS PER LITTER 3 -- PFOA: Lactatonal and Placental Transport Pharmacokinetic Studyin Rats Mean Pup Weights Per Litter `Explanatory Notes DuPont-13309 Note One pup per litter was sacrificed on days 3, 7, and 14 for blood collection. Two pups per liter (itter size permitting) were sacrificed on day 21 for blood collection. Abbreviations SD. = Standard Error SE. = Standard Error N= Number of Liters in Group A wo PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats MmGAL sae &4 0 Mseaon. Nse AILS 7&& 72%% He5a.n s z aneL a 5& s7 M5ea:n. Sie: AmAL 0a) n7 ES H5oba.n sNie MEAN PUP WEIGHTS (grams) PER LITTER GRouR: 1 DOSE: 0 MG/KG/DAY DO DAY DW Dare mara 6s.e7 9$.34 1m37 2m6a4 aass sls 3 a837 uulss 2aas 3a es 10s 157 265 ae 6osss o5.s2o 1o2s 25l.i4sa a3s40 0s 20 05 3 o5 s o3 m 5152 GROUP: XX DAO DAY3 526937 a59l7r 8817 s5s7 5o2e o1 s os is o5 so DOSE: 3 Me/KG/DAY DAYT DA DAY 21 1aas6ss0 a330a30 os1asios9s malae asess ssassi a1s2 2a61e 3183.070 os s 2: 00 ss er GROUP: XII Dwo DaY3 76.s0 10s.a6 eess 1s0s1 2% mi 0e2s3 1o0l2is o5 c os ls DOSE: Dw 1w.z0 aBslse p30 1siss o5 m 10 MG/KG/DAY Daw is maw a m21e6 saaiss 2s9l0a7 aadzs ks sl 239.s8 asnaz 51s 2 5 Rov: Iv DAO DAY3 156 a9a8 7s5s 10233 se 57 70.308 0tos o5 17 o5 s DOSE: 30 MG/KG/DAY DAY7 Dale Dara 1153 2s0a2 a"sa ssss a30s sdass awh 2a a2 126233 ms2les 4osl.es L5 oo s38s a3 s: a Mo pups. 5 DuPont.13309 A a PFOA: Lacatonal and Placental Transport Pharmacokinetic Study in Rats DuPont13309 APPENDIX I NN PLASMA AND TISSUE ANALYSIS Tw PFOA: Lactational and Placental Transport Pharmacokinetic Sudy inRats PLASMA AND TISSUE ANALYSIS EXPLANATORY NOTES Abbreviations G= NS. = LOQ = <LOQ = PP = S*D. == Gestation NoSmple Limitof Quantitation, 0.05 pg/mL. Less than limitof quantitation Posiparum Standard Deviation Insufficient for sample reanalysis DuPont13309 LN 2 -- PFOA: Lacutional and Placental Transport Pharmacokinetic Study in Rats DuPont13309 Concentration (g/mL) of PFOA in Matemal Plasma During Gestation and Postpartum Day (Dmoesee Concentration ___Average SD. 06 0 <<LooQQ oq NA <L0Q <<LLooQQ 3 1707067 853 106 854 738 0 2480935 62 2s 22358686 2102.803 30 7571.5039 7049 894 6865 81737481 \h 156 0 3 10 0 <Lo0qQ 0Q NA <LoQ <<L00QQ 1087611 1592 129% 339.04 a9924 2240 1419 3022 296.1058 3626 s8.i4e2 ss 30 79.10 78412393 28 -- - PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont13309 Concentration (g/mL) PFOA in Maternal Plasma During Gestation and Postpartum (cont'd) Dose Day (mghg) Concentration Average SD. a2 0 <LoQ <L0Q NA <L0Q <L0Q <LOQ <L0Q 3 Las uo 22 1361 271 1193 1750 10 2628 320 668 3052 3853 3251 a8 30 7850 7636 1476 702 6560 6332 10035 : A #0 3 10 <L0Q w0Q NA <L0Q <L0Q <L0Q <LoQ 865 not zn 1062 1345 082 129 - 20 2m 198 2530 23 30 5325 5430 1786 . 57 5150 7090 276 3 TT PFOA: Lacttional and Placental Transport Pharmacokinetic Study in Ruts Dupont13309 Concentration (g/mL) PFOA in Maternal Plasma During Gestation and Postpartum (cont'd) TTDay o(mmghe) Concentrat--ion Average SD. wo oq oq a0Q Na o<LqOQ 3 <9034q 009 290 1201397 1540923 0 2203485 38 207 22%5782 2 64n9n7 wo usm 683180 EN 57.98 No we 0 5 0 30 . oq a0Q Na o<LqOQ o<0qQ 09628 06 092 983358 115028 nw 2m 22621 2261368 4027035 sae 1163 5S066498 730 - Tw PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats DuPont:13309 Concentration (g/mL) PFOA in Maternal Plasma During Gestation and Postpartum (cont'd) Dose Day (mykg) Concentration Average SD. ae 0 3 10 0 <LoQ a0Q NA <LoQ <L0Q <L0Q <LoQ 1035 904 Lol 799 968 205 812 2034 884 sis 2345 2876 35.68 3197 6327 6413 Las 6286 6430 N 66.10 ~ [ PFOA: Lacttional and Placental Transport Pharmacokinetic Study in Rats Concentration (g/mL) PFOA in Fetal Plasma on Gestation Day 21 Dose Day (mghg) -- Concentration Average SD. ao 3 10 30 <Q <Q Na <<100QQ <0Q <Q 575026 588 069 553 550318 21 Laas ust 1490 1155591 Bs un Bu as 3272.079 23259960 DuPont13309 : ~N wo PFOA: Lactaional and Placental Transport Pharmacokinetic Stdy in Ras Concentration (g/mL) PFOA in Pup Plasma Day (Dmogsheg) Concentration Average SD. wo 3 10 <1o0qQ <0Q NA Log <<L0oQQ 324 28 070 159 23135 s3e56 594 tas sol 850068 3 146 19% 166 11s Ba 1038877 wo 3 <0. Q <L0Q NA 885 <Loq <0L7o8Q 06s 020 0a 005874 10 338 2m oss 232082 203 30 3a1n3 a1 . oq 561651 370 DuPont 13309 : \,. a Sample not used in calculation. Extraordinary high value likely the resultofsample contamination, - ss "PFOA: Lactatonal and Placental Transport Pharmacokinetic Study in Rats Concentration (g/mL) PFOA in Pup Plasma (cont'd) Day _D(mogsheg) Concentration Average SD. we 0 3 10 30 <Loq <0Q NA <<L00QQ <L0Q <L0Q 0o8r7 017 oo 074 0068s7 225407 22 ox 210662 244 as ast 352130 501 638 20 0 3 ooqq oq NA <<LLooQQ Log 131 128 om 013060 10 232575 325 0s 267 33480 276 0 671086 [EY 1050 570 DuPont 13309 : ~ -= PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats Concentration (g/mL) of PFOA in Milk Dose (mg/kg) 0 Average SD. 3 <L0Q <L0Q <L0Q <L0Q <L0Q <LoQ Na Postpartum Day 7 14 <L0Q <L0Q <LOQ <L0Q <L0Q <L0Q <LoQ <L0Q <L0Q 021 <LoQ Na NA NA 3 081 092 Lal 093 128 Average 107 SD. 026 015 L10 090 120 090 112 084 109 131 122 094 115 LE 006 10 216 197 221 149 234 Average 203 SD. 033 232 324 404 534 332 365 201 221 1.98 283 274 345 091 118 30 532 655 595 633 491 455 497 7.47 7.73 520 52 614 304 434 7.87 Average 497 SD. 120 576 645 126 138 a Average not calculated because 3of4 values <LOQ. DuPor-13309 21 <L0Q <L0Q <L0Q <LoQ <LoQ <L0Q NA L14 NS. Lot 117 118 113 008 a 280 372 280 252 349 307 ost 736 NS. 983 636 636 7.48 1.63 135 PFOA: Lactational and Placental Transport Pharmacokinetic Sudy inRats DuPont 13309 Concentration (ug/mL) of PFOA in Amniotic Fluid Dose (mg/kg) Day 156 Concentration __Average SD. 0 oq aoQ NA <<t0oQQ <10Q <0Q 3 0022s 060 06 013623 <Loq 10 07 060 on ois 049 038 079 30 019294 170 oot 325 112793 Day21G Concentration __Average SD. oq a0Q Na <<00QQ <<Lo0QQ 210280 [ET 112299 1.66 2S9070 376 os 450 208 398 786773 5 ose 754 703327 [3 PFOA: Lactational and Placental Transport Pharmacokinetic Study in Rats Concentration (g/mL) of PFOA in Placenta Dose Tissue Day mg/kg) Concentration Average S.D. 156 0 <L0Q <L0Q NA <L0Q <LoQ <L0Q <L0Q 3 1.48 22 am 133 sal 161 128 10 745 si 170 452 <L0Q 343 501 30 201 B20 B17 1295 1486 B14 26 0 3 10 30 <Q <L0Q NA <L0Q <L0Q <LoQ 338 355 057 288 365 321 445 690 ox 176 849 nn 1091 9.40 2985 23 an 228 27.16 1943 2.02 a One control sample was usedformethod developmen. 137 DuPont. 13309 : NNo PFOA: Lacutions! and Placental Transport Pharmacokinetic Study in Rats Concentration (g/mL) of PFOA in Embryo Day Dmogsse) ConcTeinstsruaetion Average _S.. 106 0 oq @0Q NA <LoQ <<1L00QQ <L0Q 3 L1i58 10 030 114095 176 0 249459 333 os 33599 30 H23s1 na 350 233 11805105 995 156 0 3 10 30 . 40Q d0Q NA <<LLooQQ <LoQ <0L1o9Q 024 ow 001537 001174 055 053 018 006217 048 017369 24 on 0L9a7 131 106 138 DuPont.13309 ANNo PFOA: Lactational and Placental Transport Pharmacokinetic StudiynRats Concentration (ug/mL) of PFOA in Fetus, Day Dose (mg/kg) _ Tissue Concentration Average SD. 26 0 40Q <0Q NA <L0Q <L0Q <L0Q 3 <0Q 095 21 02 165 Lis 126 135 10 - 260 037 269 279 206 30 289 223 sm 236 1228 752 307 ee DuPont. 13309 NN a Sample lost due to misinjecton. ----- El