Document MBEB6gb69XG9ny0Qoyrk1Dmj
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oDwupree1s530:2
Study Tide
Ammonium Perfluorooctanoate:
Age Effect on the PFOA Plasma Concentration in Post-Weaning Rats Following Oral Gavage
"TEST GUIDELINES: None applicable
AUTHOR: Paul M. Hinderliter, Ph.D. STUDY COMPLETED ON: December 2, 2004
PERFORMING LABORATORY: E.L du Pont de Nemours and Company `HaskellTM Laboratory for Health and EnvironmentaSlciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 LABORATORYPROJECT ID: DuPont-15302
`WORK REQUEST NUMBER: 15339
SERVICE CODE NUMBER: 1389
SPONSOR: EL du Pont de Nemours and Company
Wilmington, Delaware 19898 USA.
and
3M Company
3StMPCaeuln,teMr NBui5l5d1i4n4g-1000
mm
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APlmamsomnaiCuonmcePnetrrfautoiroonoicntaPnoosste-:WeaAngiengEfRaitcs oFonltlhoewing Oral Gavage
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GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT TLhaibsorsattuodryywParsacctiocneduScttaneddairndsc,omexpcleipatncoerwtihtehiUt.eSm.dEoPcAumTenStCeAd (b4e0loCw.FRThpaertit7e9m2)liGstoeodddid not impact the validityofthe study. + Nexopecroinednuccetd,autdriatinweadspecrosnodnuncetl.edAolnl twhoirskstwuadys,dhoocwuemveenrt,eadlliancttihveitsiteusdwyerreecopredsr.formed by
StadyDirector:
ReseMa.rcHihnTdoexrilcioelro,gPishtD.
onde 200k Date
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APlmummosniCuomncPnersfauioornoaisnaPnoosseW:eaAnginegEfReattoFnoltlhoewing Oral Gavage
QUALITY ASSURANCE DOCUMENTATION
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`Work Request Number: Study Code Number:
15339 1389
"The conductofthis study has been subjected to periodic Quality Assurance inspections. The dates of inspection are indicated below.
eee E---- ere
Phase Audited Audit Dates
BEETer
ees
Date Reported to Date Reported to
Study Director
Management
Protocol:
July 22, 23,2004
July 23, 2004
July 23.2004
Report/Records: ~~ October 28, 29, 2004 eeOctober 29. 2004 -------- November 24, 2004
Reported by:
C
JosepCh. Hamill
`Quality Assurance Auditor
L- Dec 2204
Date.
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PAlsmmaoCnoncPornfrlasioornooicnnPoosstts: cAagneiEnglRtes nFcolttlhoewing Oral Gavage
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CERTIFICATION
We, the undersigned, declare that his report provides an accurate evaluationofdata obtained
from this study.
soAuitta (uit ToCr MPaatitne,a55t. es
vic Coit
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Approvedby:= -- wH i Resc) he Mange n 3b 0 atoe v=2008
eedby Study Dito:
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TABLE OF CONTENTS
Page
GOOD LABORATORY PRACTICE COMPLIANCESTATEMENT csi
QUALITY ASSURANCE DOCUMENTATION...
mans
d
A
STUDY INFORMATION ocr]
BC nimmmmmmmm------_5
INTRODUCTION ceramic)
MATERIALS AND METHODS cco
A)
Ar TSE SUBSIANGE vrs
wl
B. TRE BI gmrmmmmmmmmmnmmm--------y
. Ail TREY wemrmmmmmmmmmmmpmmmm--11
2 Cogent Case Bark CMINR nurmmmmmmmmentmmmommmmrnmmmind1
FC
FE
------
5. Animal Health and Environmental MONOFing PROSIAM cvs |
D. PretestPeriod vv
I
--
E. ASSIEAMENL LO GIOUPS..vrrrs sss 12.
Tr ATS rm
----
ro 13
2. DosePreparation, ABaIYSiS, andRAE ..vevvvsessssssesssesninsese |
3, Saree BIER mmol
4. EXPETMENal PICNIC...
|
G. SUPPIEMENAl STAY rvs
14
En
----
1. Verificationof Dose Solution.
:
nnn14
2. ExtracotfiPFoOnA from PIasma forLC/MS ADBISSIS..eurvevrrvsnrre 14
3. SAARI mommmmmmmi------"ht
4. Chromatographic Methods. ...........rowrvmerrmr
ent
I SUSHCAl ANGIYSES rvs 16
RESULTS AND DISCUSSION .ccorrsnssssssnssmmsnsssssssasssmsssnnson1n6
J FE
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37 WIE REC cnssmmmmsmmmm----------
16
2. FemaleRats...
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esmnig--_0
3. SupplementalRatS vv...
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C. Urine CONCENTAtion Of PFOA ..vvvvresvsssenessnsonesnasensnnnn 1
CONCLUSIONS rmsd]
RECORDS AND SAMPLE STORAGE ..cvcersesssmsonsmsmsnessssmsnssmsmonorsono18n
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Table 1:
Plasma and urine 30 MEKEPFOA
concentrations in male rats oral gavage with 10 or cress
22.
Table 2:
Plasma and urine concentrations in female 30 MEAGPFOA rrrrnennsrnss
rats oral gavage with 10 or sess
23
TITFEiguRre,1:cccPularsommasceounscemnstsramtpioons in rats folln owing oral gaa vage With Pg FOiAn.c.tcv.crs veruvon. 25
Figure 2: Urine concentration in ratsfollowing oral gavage With PFOA ....c..........26
APPEADNPDEINGCiExSA:coDOrSvIrNrGeSsOenMsOmsAsDmsRsmIssYsSssIssSs.ss.nsvssrsvssrssssosmsssosssmssmsssns:ond2]9
`Appendix B: Plasma Concentration Da-MAtlESa vn]
`Appendix C: Plasma Concenration D-aFEtmMaaIES.......vvorvvovsrsesnsns 44
`Appendix `Appendix
D: E:
Supplemental Pasi Urine Concentration
DCONaG-EMNtTaAtalONeS s.......vvovvsivronnowndd)T
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STUDY INFORMATION
9th Collective Nomenclature:
Synonyms/Codes:
Octanoic acid, pentadecafluoro-, ammonium salt
++ FACm-m1o4n3iFuLmUpOerRfAluDorBoroactnadnoFaltueorochemical Surfactant
(3M Company, Specialty Materials)
+ C8
++ APePrFflOuorooctanoate, ammonium salt
+ PFOA
+ HLo-t24393221(3M Specialty Materials) (Lot No.)
HaskellNumber: 24921 `CASRegistryNumber: 3825-261
Purity: 95.2% - 97.99%
Straight chain: 77.6%
Branched: 12.6% internal monomethyl (non-alpha)
9% isopropyl
0.2 % tert-butyl
00..11%% galepmh-adimmoentohmyelthyl
Known Impurities: C+(CsF,COy' NH"), 0.01%
C5(C4FsCOy NHL), 0.03%
CCo1((CCasFFinsCCOO;y NNHHLL)),, 005473%% Co(CFinCOy NHL), 0.16%
Monohydro APFO, 0.09% Monounsaturated APFO, 0.72%
Undefined (possibly) substituted perfluorocyelo species, 0.2% cyclopentyl, and 0.1% cyclohexyl
Physical Characteristics: White solid
Stability: The test substance appeared to be stable under the
conditionsofthe study; no evidence ofinstability was observed.
Study Iniiated/Completed: July 8, 2004 (ce report cover page) Experimental Start/Completion: August 9, 20/O0ctob4er 13 2004
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SUMMARY Perfluorooctanoate (PFOA) plasma concentrations were measured in immature (3, 4 or 5 weeks ofage) rats following single oral gavage doses at 10 and 30 mg/kg. Plasma concentrations varied significantly between experiments, especially when compared to a prior study with 4-week old animals. In the current study, male rat PFOA plasma concentrations are relatively constant in `male rats across age groups, varying only with dose level. Female rat plasma concentrations decreased with increasing age, but was only statistically significantly at the 30 mg/kg dose level. PFOA concentrations in urine, while not normalized for total elimination, indicated more rapid urinary excretion in female rats at all ages. The plasma concentrations from the supplemental `group, however, are significantly different from the main study levels for both male and female: rats. The plasma concentration variability in 4-week old rats suggests an unknown physiological variability in animals younger than 5 wecks. As a result, the relationship between age and PFOA plasma concentration is undefined in 3-4 week-old ats following oral dosing with 10 or 30 mg/kg PFOA.
A Phsma PFOA plasma concentrations in male rats were not significantly different by sample time (at 2 and 24 hours)or by animal age (3, 4 or 5 weeks) at either 10 or 30 mg/kg, exceptat 2 hours and 5 weeksofage. The PFOA plasma concentrations in female rats were significantly lower at 24 hours post dosing (compared to 2 hours) at allagesand doses tested. Female plasma tcoinmceesnatnradtdioosnseswe(pre<0a.l0s1o)l.owTehrefPoF5rOwAeepkl-aoslmda rcaosnctehnatnra3tiaonnsd4fowleleokw-ionlgdar3at0s matg/bkotghdsoasmeplwee:re approximately 1.5 to 4 times higher than those observed following a 10 mg/kg dose. Ipnoast-pdroiosrinsgtufdoyl,lmoawilnegaandsifngelmeal1e0rmagt/PkFgOoAralplgaasvmaagecodnocseen.trPatFiOonAs pwlearsemmaecaosnucreendtraatti2o4nshoaurres consistentwith the current study for both sexes at 5 weeksofage but not at 4 weeksofage (3swteuedikeso,ldinadniicmaatilnsgwtehratebnootthparreeviionutselmyalmleyascuonrseids)t.enTt.heThsteanodnalrydkdneoviwantivoanrsiaatrieonsmbaeltlweinenbotthhe two studies is whether the rats were fasted before dosing. The supplemental rats were added to ienxcarmeianseedtthheecPffFeOcAoffpalsatsimnagcoonnctehnetrPaFtiOoAnspalsaesxmpaeccotnecdebnuttratthieonnsomiinn4a-lwevaelkesoladreradsi.ffeFraesnttifnrgom cither the current study or the prior study. Given the consistency in the 5-week old rat PFOA pyoluansgmearcotnhcaenntrwateieoknss,WhitiacphpeiamrpsactthasttthheerpehairsmaancoukniknnetoiwcnsopfhysPiFoOloAg.ical variability in rats
B. Urine TFheemraelewerratesssihgnoiwfeicdanhtigdhiefrfePrFenOceAsuirniPneFcOoAnceunrtirnaetcioonncse,natnradttihoenssewiintchreaagsee,ddwoiset,h aagned.seMxa.le rats showed the opposite, with urine concentrations decreasing with age. Both sexes showed higher
ER
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m(2a.d5-e6.t5o tqiumaenst)ifuyriunreincaornyceonuttrpautti(ovnoslautme3)0,magn/dk,gbeccoamupsaercerdeattoin1i0nemgl/evkeglsdwoseerse.nNotomaetatseumrpetdw,as adjustments for urine concentration could not be performed.
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INTRODUCTION Ammonium perfluorooctanoate (APFO)is a synthetic perfluorinated octanoic acid used as an industrial surfactant. Eliminationofthe disassociated anion, perfluorooctanoate (PFOA) in rats is sex-dependent and appears to be under hormonal regulation. PFOA elimination is much faster in female rats than in male rats, down-regulated by testosterone in both female and castrated male rats, and up-regulated by estradiol in the mal rats. A prior study conducted at DuPont HdiafsfkeerlelncLeabwoarsatloerasytfaotu4ndweseekx-sdoefpaegned,enatt tPiFmOeApeerliiomdijnuasttiopnriionr 4{-08swexeueakl omladturraattsi.o)n Tinhethseerxa. `The objectiveofthe this study was to determine if3-week old male and female rats eliminate: PFOA from blood ata ate differen than that observed in 4- or S-week old rats. Male and female post-weaning rats at different ages were dosed with APFO, and the PFOA plasma concentrations were compared a2t and 24 hours postdosing. At24hoursafter dosing, the sexdifference in PFOA elimination in sexually mature rats was easily distinguished via the plasma concentration wofasPdFeOsiAgn.edUtroineevawlausatceollilmeictteedd kfionre2ti4cheonudrpsoianntdsaenxacllyuzseidvefoorfPdFetOeArmicnonacteinotnroafteiloni.minTahtiisosntruadtye constants
MATERIALS AND METHODS
A. Test Substance APFO was obtained from 3M (St. Paul, MN) and assigned Haskell Laboratory Number H-24921 upon receipt. Available information on the purity, composition, contaminants, synonyms, hazards, and hazardous material classifications) were provided by the vendor and documented in the study records and/or report.
B. Test System MRaalleeigahn,dNfoermtahlCeaCrorlli:naC.DA(nSiDm)aIlGsSfBoRr grartosuwpserIeVobtaaririnveeddfarsomlitCehrasrwlietshRdiavmesr.LabTohreatSoprriaegs,ueI-nc., Dawley rat was chosen for this study becauseofthe extensive experience with this strain and its suitability with respect to longevity, sensitivity, and low incidenceof spontaneous discascs. Furthermore, the Sprague-Dawley rat has been used previously for toxicokinetic testing of PFOA
and other fluorinated test materials. **
At the timeofdosing, rats were approximately 3,4, and 5 weeksofage and the weight variation diindclnuodtedextcheeeHdask2e0ll%aonfimthael mneuamnbewreiagnhdt abnyidnodsievigdruaolupidaenndtisfeixc.atiIonnfnourmmabteironasosnigthneedctaogecalcahberlast. `The individual identification number was placed on the tailofeach rat by tattooortail mark.
Thos
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APlmamsomnaiCuomncPeenrrlaotroooncitnaPnoosatt-eW:eaArgiengEfRfeactt oFnotlhloewing Ora Gavage
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C. Animal Husbandry
1. Housing
Upon arrival at Haskell Laboratory, rats were removed fiom shipping cartons and housed in appropriate cages according to Standard Operating Procedures unless specified in the study records. Rats were maintained under quarantine for at least three days unless approved otherwise by the site veterinarian, had at least one recorded weight gain, and no abnormalities detected. After the quarantine period, ats were selected for study.
During pretest, dams were housed with their litters in polycarbonate pens containing bedding. `Throughout the dosing period with test substance, the rats were housed individually in appropriate cages according to the SOP. During the urine collection period, rats were housed in stainless steel wire mesh cages.
2. Cageand Cage Rack Change
Due to the lengthof the study, no cage and cage rack changes were required.
3. Environmental Conditions
Animal rooms were maintained at a temperature of 18-26C (targeted to 22-24C) and a relative humidityof30-70% (targeted to 40-60%). Animal rooms were artificially illuminated (fluorescent light) on an approximate 12-hour light/dark cycle. Unlessjudged by the study director or the laboratory veterinarian to have significantly affected the resultsofthe study, the relative humidity and temperature ranges in the housing rooms were recorded but were not included in the final report.
4. Feed and Water
Al rats were provided tap wateradlibitum and fed PMINutrition Intemational, LLC Certified
Rodent LabDiet 5002ad libitum. Rats were not fasted prior to dosing due to their age.
5. Animal Health and Environmental Monitoring Program
As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures were performed periodically to ensure that contaminant levels were below those that would be expested to impact the scientific integrityofthe study:
+ Water samples were analyzed for total bacterial counts, and the presence ofcoliforms, lead, `and other contaminants.
+ Samples from freshly washed cages and cage racks were analyzed to ensure adequate sanitation by the cagewashers.
reCeqrutiirfeimedenatnsiamanldfneoetd twoaesxucseeedd,sgtuaatreadnmtaeexdibmyutmhecomnacneunftarcattuironesrotfokmeeyetcosnpetcaimfiineadntnsu,triitnicolnuadling
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specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presenceofthese contaminants below the maximum concentration stated by the manufacturer were not expected to impact the integrity ofthe study.
"The animal health and environmental monitoring program was administered by the attending laboratory animal veterinarian. Evaluationofthese data did not indicate any conditions that affected the validity ofthe study.
D. Pretest Period
Upon arrival at Haskell Laboratory, all rats were housed in quarantine. The rats were:
+ quarantined for at least 3 days.
+ identified temporarily by cage identification.
+ weighed at least 3 times during quarantine and once prior to initiationofexposures.
+ observed with respect to weight gain and any gross signsofdisease or injury.
The rats on body
were released from quarantine weights and clinical signs.
by
the
laboratory
animal
veterinarian
or
designee
based
wRiatthsotuhtatnewcerroepsayc.cidReanttsaltlhyatKwilelreed ofrournedmodevaedd ofrrwomersetusdaycrdiufriicendgitnheexptrreetemsitspdeurriiondgwtehreeprdeitsecsatrded pienrcilouddewdeirnetghievfeinnaagrreopsosrtexunalmeisnsatcionosnitdoercehdecskigfnoirfitchaentprteostehneceeovafludaitsicaosneo.ftThheesrteusduyl.ts were not
E. Assignment to Groups Rclaitnsicwaelrseigsnesloefcdtiesdcfaosreusoeroinnjusrtyu.dyTbhaesyedweorneaddiesqturaibtuetebdodbyy cwoemipguhttegraiizneda,ndstfrarteiefideodm from any rstaantdisotmiiczalaltyisoingniinftiocsanttuddyifgfreoruenpcseassadmeosinggnagtreodupinbtohdeySwteuidgyhDtemseiagnns,.soRtahtast ftohrergerowueprseInoV were eaxlscoeerdan&do2m0i%zoedftbhyedmaemantoweenisguhrt,e eTvhene dfiisrsttriSbuattisoni.n Teahcehwgerioguhptwvearrieatdieosniogfnsaetleedcftoerdsraactrsifdiicdenaott the 2 hours and the remaining rats in each group were designated for sacrifice at 24 hours Eidaecnhtirfaitcawtaiosnansusmibgenred. aTuhneiqluaset63-ddiiggiittsHoafstkheellHaansikmealll nanuimmbaelrnaunmdbeanr iwnadsivtiadtutaoloecadgoe:n the tail of ceaagceh rlaatb.el.The Haskell animal number and cage identification number were both included on the fRoartsotthheart lwaebroerantootryaspsuirgpnoesdestooratesasctrigfriocuepd,biyncclaurdbionngdtihoexdidaemsasfprhoyxmigartioounpsanI-dIVd,iswcearrederdelweiatsheodut anatomic pathology evaluation, at the discretion ofthe study director.
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F. Main Study
1 Animals
"The experiments were composedofthe following animal groups:
Mal"eGrowFemale (Dmopseg) Weeksre Days MNaleumoFrbeRmaeasl'er_
m' own 1500 33 aa wwoo 1100 vvii vvmt 1500 34 3EE 10 1100 Ixx oxx 510 55 3 B0 w 1100
Adnoisimnaglwawserdeoc1umdeanytefdroinmtthheesatrugdeylraegcoradtsthaendmloer foifndaoseipnogr.. The aca 3301 "GSisveenngrasoupsisnagclreifgiacevdagaied2osaen:d 24 hours pos-dos for blood collection.
2. Dose Preparation, Analysis, and Rates
`The test substance was administered as a single oral gavage dose. This route was chosen because itis the route most commonly used for toxicity studies ofAPFO. APFO was mixed with NANOpure water to achieve the target dose concentrations. Dose levels of 10 and 30 mg APFO/kg body weight were used with a dose volumeofapproximately 4 mL/kg body weight.
Dose was prepared on the dayofuse or prior to the dayofuse and stored under appropriate conditions
HPLC-MS methods were used to confirm PFOA concentration in the dose solutions
3. Sacrifice and Blood Sampling
Rats were sacrificed by CO; asphyxiation and blood collected from the vena cava or by cardiac puncture.
4. Experimental Procedure
Rats were obtained from the vendor and administered PFOA at dose levels described above. Two hours after dosing, $ rats/sex/group were sacrificed and whole blood collected. Whole blood samples were placed into EDTA tubes and maintained on wet ice. Plasma was separated by centrifugation and frozen at <-10C uniil analysis.
"The remaining 5 rats/sex/group were placed in metabolism cages for urine collection. Urine was collected for 24 hours after dosing. At 24 hours, all remaining rats were sacrificed and whole: blood collected. Blood was handled and separatedasabove.
=
APlmamsomnaiCuonmcePnetrrfaltuioroonocitaPnoosntte-:WeaAngiengEfRfeacst oFonltlhoewing Oral Gavage No rats were found dead or were sacrificed in extremis.
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G. Supplemental Study Four additional male and four additional female rats were added to groups V and VI. Two ofthe wmiatlhetaenstdstuwbosotfantchee. fDeomsaeleprreaptasrwaetrieonfaasntdedadomvienrinsitgrhattifoonr awpapsrtohxeimsaatmeelyas12inhtohuersmabiefnosrteuddyo.siAnlgl rats were sacrificed at 24 hours post dosing and whole blood collected. Blood was handled and scparatedas above. HPLC-MS method was used to determine plasma concentrations ofPFOA.
H. Analytical Methods 1. Verificationof Dose Solution
Methods for verificationofdose solution are detailed in Appendix A. 2. Extraction ofPFOA from Plasma for LC/MS Analysis
Pprleacsipmiatastaimopnlceoslwuemrnes p(rJoocneesssCehdrboymaptrootgerianpphrye,ciLpaiktaetwioonod(,PPCTO)).usAing0.s5olpugt/emALrrsaolyutpriootneoinf apnerifnltueomraolnosntaannodiacrdacfiodr (quAalndtriitcahtiCohnemoifcPaFlsO,A.MilPwlaauskmeaes,amWpIl)esinwaecreteontihtarwieled,(AaCndN)a w20asiuLsaeldiqausot oafppcraocphrisaatmepdlieluwtiaosnarpapteliveodltuomethseoPfPATCNa/riranyt.erTnhael sptlaansdmaardsasmoplulteisonwetroethperePcPipTitaartready.byDaidldutiinogn srtataensd,arradnsgoilnugtiforn)o,mw1e:r1e0 (ut1i8l0izLed oinfoirndteermtaolcsatpatnudraerdthseolsuatmipolne) tcoon1c:e2n0t0ra(t3i9o8n0s wpiLtohfinitnhteemsatlandard cceunrtvreifpuagreadmeatte~rs3.00T0hrepamrrfaoyrwSamsinsultoewsl,yaenludteexdturancdtesrwvearceuaunmaliynztoedab9y6-LwCe/llMSr.eceiver plate,
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3. Instrumentation
System: MDoetdeect:or:
SourcLeM: 1 resolution: HToMn e1nreersgoylu1t:ion EConlrlainscoen:: ELxMit2 resolutions HToMn 2enreesroglyut2:ion: MCaapliilllaireyr((kVV)y: CExornaec(iVo)r:(V): RSEourlecnes t(eVm)p:erature:
MSDeMestohloadt:iontemperature: TMiomdee:: chi
an:
Wheaatteerr,sn27d95auLtioqsuaimdplCehrromtogrsph, equipped with quserary pump, coun QMuRaM Micro Mass Spectrometer N1e0g0ative Electrospray 11000 2s 51 210 020 1L5o o0c 350C M0-R1M0,0 m2iannsitons 4Du1e3l.l0:0 369.00 02550 C4o6l3ls0i0on5en4e1r9g.y0:0 15V DCaulellison energy: 01526575
4. Chromatographic Methods
CMoeltuhmond:: CMoobliulmenptheamspeesr:ature:
PWlasamaXcitoenrcreaenMirSartCo1n8s,an2a.ly1sxis30mm2.,5 im AA:mbi50enmtM Ammonium Acetate B: Acconiile
Gradient:
[mem Tp]
--ooTs
os
Te
eo
1 0--]
ee
00]
Che Tw 1
FStloopwiramtee:: Injection volume:
01205mmiLnmin Si
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L Statistical Analyses
Group data is represented as Mean + SD. Where appropriate, statistical significance was assessed by the Student's test
RESULTS AND DISCUSSION
A. Dose Solution Analysis (Appendix A) `The concentrationsofth test substance in the dose solutions were verified by LC/MS method as described in MatearndiMeathlodss, Section F. Standards for dose verification and the dose: solutions were prepared by different personnel. The determined mean PFOA concentrations of the dose solutions were all within 20%ofthe targeted concentration (2.5 and 7.5 mg/mL, Appendix A). PFOA was stable in the vehicle foar least 5 hours.
B. Plasma Concentration ofPFOA (Tables 1-2, Figure 1, Appendices B-D)
I. MaleRats `The PFOA plasma concentrations were not significantly different by sample time (at 2 and 254wehoeukrsso)foargbey(apn<i0m.a0l1abgaes(e3d,4onotrhSe wreeseukltss)aofueniptahierre1d0to-rte3st0amtg9/9kg%,ceoxncfeipdtenacte2ihnoteurrvsal)a.ndIn some groups the 24-hour post dosing concentrations were higher, but this was not unexpected dcounecetnottrhaetiloonnsg rpalnagsemdaferloimmi2n1at.i0o0ntoofP48F.8O5Apgin/mmLalfeorrtahtes.1)0 Imngd/ivkigdudaolsePFanOdAfprloams4m9a.51 to w1e5r3.e8a9pgpr/omxiLmaftoerltyh2e-330tmigme/skghidgohseer.thPaFnOtAhospelaosbmsaercvoendcefnotlrlaotwiionngsafo1l0lomwgi/nkggad3o0sem.g/kg dose
2. Female Rats P2 FhoOuArsp)liansamlal caognecseanntrdadtoisoenss wteesrteeds.igPniFfOicAanptllyaslmoawecronatce2n4trhaotuirosnspowsetredoaslisnogl(ocwoemrpaatr5edweteoks tfhoalnlaotwi3ngoa4r30wemegk/skgatdobostehwsearmepalpeptriomxeismaatnedlydo1s.e5st(op4<ti0.m0e1s).higPhFerOAthpanlatshmoasecoonbcseenrtvreadtions following a 10 mg/kg dose.
--_--
"o7
APlmamsomnaiCuomncPeenrrflautoornoocintaPnoosatteW:eanAignegEfRfetcst oFonltlhoewing Oral Gavage
Dupont-15302
3. Supplemental Rats
Individual PFOA plasma concentrations in the male rats were 64.95 and 30.00 pg/mL for the 6fa8s.t1e6daanndd2n6o.n5-4faustge/d Lanifmoarlts,hrfeasspteecdtiavnedlyn.onP-fFaOstAedplaanismmaalsc,onrceesnptercattiivoelnys.in the female rats were:
Iponsatpdroisoirnsgtufdoyll,o"wmianglae
and female rat PFOA
single 10 mg/kg oral
plasma
gavage
concentrations were
dose. PFOA plasma
measured at 24
concentrations
hours
are
consistent with the current study for both sexes at 5 weeksofage but not at 4weeksof age (3-
`swtuedeikeso,ldinadniicmaatilnsgwtehratebnootthparreeviionutselrnyamlleyascuonrseids)t.entT.heThsteaonndalrydkdneovwiantivoanrsiaatrieonsmbaeltlweinenbotthhe two
estxuadmiiesneistwhheeetfhfeecrttohfe fraasttsiwngeroenftuhsetePdFbOefAorpeldaossmiangc.oncTehnetrsautpipolnsemiennt4a-lwereatks owledrreatas.ddeFadsttoing
ieintchreerastheed ctuhrerPenFtOsAtupdlyaosrmtahecopnrcieonrtsrtautdiyo.nsGaisveexnptehcetecdonbsuitsttehnecnyoimnitnhael Sv-aulweesckaroelddirfafterPenFtOfArom
`psloausnmgaercotnhcaenntrwateieoknss,whitiacphpeiamrpsactthasttthherpehairsmaancoukniknneotiwcnsophfysPiFoOloAg.ical variability in rats
C. Urine Concentration ofPFOA
(Tables 1-2, Figure 2, Appendices E-F)
PhaFdOhAiguhreirnePFcoOnAcenutrrianteicoonnscdeinftfrearteidosnisgtnhifaincamnatlleyswiatnhd atghee,fdeomsaeleanPdFsOcAx (upri<ne0.c0o1n)c.enFteramtailoensrats
iagnec.reaBsoetdhwsietxhesaghe.adMhailgeherra(ts2.s5h0ow6e.d5 tthiemeosp)poPsFitOeAwiutrhinuercionencceonntcreanttiroantsioatns30demcgre/aksgincgowmiptahred
1010 mg/kg doses. creatinine levels.
No
attempt
was
made
to
quantify
urinary
output
(volume)
or
standardize
for
CONCLUSIONS oPfeargflou)oraotosctfaonlolaotwein(gPsFiOnAgl)epolraalsmgaavcaognecednotsreastiaotns10wearned m3e0amsgu/rkegd.inPliamsmmaatucroenc(e3n,t4raotrionwseveakrised sainginmiaflisc.anItnlytbheetcwuererenntexspteurdiym,emnatsl,eersatpePcFiaOlAy pwlhaesnmacocmopncaernetdrattoioanpsriaorre srteluadtyivweiltyhc4o-nswteaenkt oilnd dmeaclreearastesdawcirtohssiancgreegarsoiungpsa,gev,arbyuitnwgaosnloynlwyitshtadtiossteicalelvleyl.sigFneifmiaclaentrlaytaptltahsem3a0cmongc/ekngtrdaotsieonlsevel, PurFinOaAryceoxnccreenttiroantiionnfseimnalureinrea,twahtiallel nagoetsn.orTmhaelipzleadsmfoarctoontcalenetlriamtiinoantsiofnr,oimndtihceasteudppmloermeenrtaaplid rgartosu.p,Thheowpelvaesrm,aacroensciegnntirfaitcianotnlvyadriifafbielrietnyt irno4m-wteheekmoalidnrsattusdsyuglgeveesltssfaonrubontkhnomawlnepahynsdiofleomgailceal pvalraisabmialictoyncinenatnriamtailosn yisouunndgeefritnheadni5n w3e-4ekwse.ekA-osaldrreassulf,otlhleowrienlgatoiroanlshdiopsbiengtwweietnha1g0eoarnd PFOA 30 mgkg PFOA.
--
APlmamsomnaiCuonmcePneiralutoiroonocintaPnoosnte-:WeaAngiengEfRaftescoFntoltlhoewing Oral Gavage
Duponc15302
RECORDS AND SAMPLE STORAGE
A sampleofthe test substance will be collected for archive purposes and retained at Haskell
Laboratory, Newark, Delaware. Specimens(if applicable), raw data, and the final report will be
retained at Haskell Laboratory, Wilmington, Delaware.
Newark,
Delaware,
or
at
Iron
Mountain
Records
Management,
-uw.
APlmamsomnaiCuonmcePneirrlautoiroonocitnanPoonstic-:WeaAngiengEfRaetcs oFonltlhoewing Oral Gavage
DuPont15302
REFERENCES
1. Ohmori K., Kudo, H, Katayama, K., and Kawashima, Y. (2003). Comparisonof the: toxicokinetics between perfluorocarboxylic acids with different carbon chain length. Toxicology 184, 135-140.
2. Vanden Heuvel, 1.P., Davis, 1.W., Sommers, R,, and Peterson, RE. (1992) Renal excretion
ofperfluorooctanoic 7,31:36.
acid
in
male
rats:
inhibitory effectof
testosterone. J.
Biochem.
Toxicol.
3. Kudo, N., Suzuki, E., Katakura, M., Ohmori, K., Noshiro, R. and Kawashima, Y. (2001) T`eCnogmtpharinisroatnos.fCthheem.eliBmioiln.atIinotnerabcet.twe1e34n,p2e0r3fl-u2o1r6i.nated faty acids with different carbon chain
4. tYhleinuerni,naMr,y eHxacnrheitjiaornvoif,pHe.r,flJuaoarkoooncathaon,oJi.c, aacniddPienutrhae, mP.al(e19r8at9.)PShtairmmualcaotli.onTboyxioceoslt.ra6d5i,ol27o4f277.
5.
DPluaPsomnat CHoansckeenltlraLtaiboonraitnoProys(t2-00W3e)a.niAngmmRaotnsiFuomllPoewrifnlguoOrroaolctGaanovaatgee:.
Age Effect on the. Unpublished report,
DuPont-13267.
6.
DuPont Haskell Laboratory (2003). Unpublished report, DuPont-7473.
Perfluorooctanoic Acid:
Toxicokinetcs in the Rat.
7.
DuPont HaskellLaboratory.(1997). Exposure. Unpublished report.
Hazard
Characterization
for
Human
Health
C8
8. dViasntrdiebnutiHoenu,vmeelt,aJbPo,liKsums,liaknids,elBi.m,inVaatinonRoaffpeelrgfhleumo,roMo.cJt,a,naoincd aPceitderisnonm,alRe.Ea.n(d19f9e1m)a.leTriastss.ue J. Biochem. Toxicol. 6, 83-92.
-es "+6
APlmamsomnaiCuonmcePnetrrlautoiroonocitnaPnooastte-:WeaAngiengEfRfaectt oFonltlhoewing Oral Gavage
DuPont15302
TABLES
-
APmlmsosmnaiCuonmcPeenrtfrlautsiroonociinaPnoosaite-:WeaAngiengERfatfsoeFnocltlhtoewing Oral Gevge
TABLES
EXPLANATORY NOTES
ABBREVIATIONS:
sD
standard deviation
DuPone15302
-_--
PlAamsmmeariCounmcePnetrraltuioornooicnaPnoosatt-eW:eaAngiengEfRfaetcst oFonltlhoewing Oral Gavage
Dupont15302
Tablel: Plasma and urine PFOA concentrations in male rats dosed via oral gavage with 10 or 30 mg/kg PFOA
Group (wAetekes) (Dmogse) v! 35 110 x 5 10 vimt 33 3500 xt 5 3
Dut expressed as pg/ml. _ZHoursPostDosePlasma_24Tiours PostDoss Mean SD Mean SD
w39g2 aadms w32 68%
u2m9 753m 4060 36
674s0 11287180 6566 155
dTeolss 1183215s las 2336
34HourTsriPnoestDose Men SD 0a5s7s a2ss am 236 2587760 2888s6 1565 624
_----
APlmamsomnaiCuonmcePnetrrlautoiroonocintaPnoost-eW:eanAignegEfRfetcst oFonltlhoewing Oral Gavage
DuPonc15302
Table2: Plasma and urine PFOA concentrations in female rats dosed via oral gavage with 10 or 30 mg/kg PFOA
Age Dose Growp (weeks) (mghe)
v1 5a 1100 xs 1 wmooo 5 3300 xt 5 30
DataexpressePdlasasugm/aml. _THousPosthose 24 Hours Post-Dose Mem SD _ _ Mean SD
8 ser msmios Bn 2a
pmsess 370 136 08
8B08667 110501 S690 066
2S80L1 193%61 se les
34 ourTsrPionsetDose Mea sD 22246 1895s2 wm 246 M94oski2 22682967 mie sis
=
"4/4
APlmamsomaniCuonmcePnetrrlautoiroonocitnaPnoosait-e:WeaAngiengEfRfaetcst Foonltlhoewing Oral Gavage
DuPont 15302
FIGURES
-_--
Ere
APmamoiniCuomnsPeertfiuoronoisntPanteo, Asge-BReasWcFtoleotlhonewiangOnraliGanve
Duron15302
Figure: Plasma PFOA concentration in rats following oral gavage with PFOA
wr
o.
.
i
-
WN \
be
w
NLL NE 1B)
22 o= m
ep
=
RNeEsR
RN NB RE
= = N k NRYk Ss BZ NNKE RE2 NANdB
|
c
i2 I.
&
Ry
[N
J5
1
T
0 2 hours post dosing
24 hours post dosing
9
1
N N
oe]
WgN
4
BES ALNem NNN
:
1
I
t
Age (weeks)
aMaleplasmaat 10mg/kg(groups, V andIX).
bMale plasmaat 30 mg/kg (groups III, VII and XI).
Female plasma at 10 mg/kg (groups II,VIandX).
d Female plasatm3a0 mg/kg (groups IV,VIIl and XII).
-------------- Fi
eres
Amos Pecaensy.Aog Elong rdGe
oisn
Figure 2: Urine PFOA concentration in rats following oral gavage with PFOA
ns
50 a
b
2 ws
T oe
5
5
g<0
mm B BB B
EiI"
5 BBIOoB
52 "
BBBsRIINNNoGE
18 Bl
S LB BBY
PISST:oEaS cIf RINToN oN .
male
BXXR) female
Age (weeks)
a 10 mg/kg (groups I I, V, VI, IXand X).
30 mg/kg (groups HL, IV, VIL, VIII, XI and XII).
-
I - 74
Co
APlmamsomnaiCuonmcePnetrrfautciroonocitnaPnoosatt-eW:eaAngienEgffReacttoFnoltlhoewing Oral Gavage
DuPont15302
APPENDICES
-
APmlmosniCuonmcePnetfruaotrioonocitnaPnoastte-:WeaAngiengERfatfsoeFnoclhlteowing Oral Gavage
APPENDICES
EXPLANATORY NOTES
ABBREVIATIONS:
L0Q
limitof quantitation
sD
standard deviation
SE
standard error
%Cv
percent coefficient of variation
Dupont15302
---
AummmseConncPeeariuroondiannPoorss:W eAg EgRfalcFtolalhnoewing Or Gage
Duronissnz
AppendiAs
Dosing Solution Analysis
--_--
A206
PAlmsmmoaniCuonmcePnetrrfaltuioornooicniaPnooates: tAg-eWERefieaFononliltohnewigng Ora Gavage
Duponiss0z
Amoi Pecfors"eWceaasnoianeg:RaAtgsFllofwicnagnOtraelPGlaarnaageConcsnraftniaotn
ANALYSISFORH-24921INDOSING SOLUTIONS
MHeateyilciSaRueRpsesgeaoNhuNPmrubomejberec:rtN:umber:
215439 Duron15302.
SIMMARY
DCooosulritnregodoolfseoioinmfsoortpnoitcyeooefnAarmignxauisns/oe,c2n0.c)05,e4,iiIon7ns3vimoelnni, 0nomfgnad4.5s2acboinvlveyoel)sprnesppare5dvansh
hitedfor Concenaions of
iswithbe ofsls. 24921 iseptdosingsuionswere
esseby igh poms
Tihesac pension eo iletfoer tenemstsyo wpacsNoasgn ar 0LwCMa rSS)
a heesblafonrH-w04a92a1snp inc(OV es oi 10) 5
ebpleredepcarveodisndlcceoewhlne(e=n 1o6nl.dA9g% ouhfsopto5,r2ar0lo0,unieqcuipaoleldsytatte
fratel.
125005ves ondetect he mn oni smpes.
Tos
APmlmaosmnaiCuonmcePnetrrfatoiroonoicnaPnoosneW:eaAnignegEfRaetscFottlohoweing Oral Gavage
Dupont15302
Dreisnpure
SAMPLESUBMITTAL
cDooooimlngtoploeudawonns stocnoAnocnfemnaiaiotnn2s/0,oo0fn.2c51e, ar0iidoo7n3,mep0cmof.aonf. ecd2o4s9oa2bl1ivsleyucpraelvypsaisss(stsiodeute
ormlswith ostof mp. `Sunplessubmitedfo nly wer nly
heday hywererecivedandlorwhen'e-
anal wasidicaed.
hespin hicefor esdwasNespaswae.
METHODS 1. Antoinette RecoverySampleAus (CN{aocsuoonrfparnettwhiitahidy)iwneigsmslttoedhts.eAnsooswck,somvelo(v2o.y5mooffL3)429492n21d1watsothpmeeppgakh dviveniNci7s.eo5pmuryend NSvpiipen,ooFinosawallaylteo1.rn2.e5Anilmrlgarteoocwno)v,eereylsdsu3.p715sempwgeorer5eenfmihqsueotdtotofrshiiesspleruitvioaonsnoffdo dbheadcis0nt1m0a0riia.of iGnoihngiuclee.sTahsmsialmpelceowncanrssihonsp.ressedndsnyzed a hesme anger
. Ding SupmionTrt GBNauciihdpotisgnwHaa2m4ep5l12e)1(0i5nahipel)ipweeanssciobocencn.iTah0ioe1nd00oimLgs.wspaimtrphlNeeasswyoprre00t0hwe0ermelgaind.dmirwsiietdh
ninadyo.)waesforaed l0tehitssa,mhe e0 gav sacqnd ndfiae OconnceLn ploefGheilical Sid thma nlspi. `hSunmpea ssubni wayfsonrsis yss. weeanya heday osloweseeived ndox
Sis
APmlmaosmnaiCuomncPeenrtfrlaotrisooncitnanPooastte-:WeaAngiengERfatfsoeFnocltlhtoewingOral Gavage
DuPont-15302
Driver CuomstogmohicCondicns
pLCrolei pr ZWoatteaArRlX2,i925n1PmLemC. 50mm,
Fiow Rae:
smomma
ConljecimomnTVeomlpumeer:s: o35uC
CMoobliulmePShawsie:ck. O1m55nniets AcidAceiie
GnTide2) ia) %Acswonicile
051500
50
5a1s
55
TMYosietminuske: MEiclroncQruEaStarDocepMpiucriaovTeayonnd.emMS
CCoopelVeaylvuoeu: biy)r
SSDeoaeuncsmTocerpiTe:rempe: P315E200ON:413ie ret o369wsdug)
T"Ch PROS 413nf arn(0370(sgh)
RetainTineofPROA1d 1.24 "CPFOA:ppro3.3 nmieis 0m0Liotilonys)
4 CabsondnQuaintiotatinon ecsttoonciklseo.luAtiponpoofpAenialiytiicqalotStoafndtaherdso(AcPkFwOe,reHi-2i27d032wi6t5h,N1m0o0p%urpagwe) ewasm(aodaekisn Bcelfiotrreatthieosensaalniqduaortwheartbarcoukgehd1 hveoulrugne,conspcperiopariiontnfamoeundtoifdmsepml asnodlatron(s1.2-
C pefuorosocic)waasnaoedi. Anpyeiciorf o(2Lm4C9e2M1StwSars)y,byNNeihg-aptcifoomnmcaeneclriwqpuirdychi roemdsttogoopearphynfeaganmtidaveselmy chrgsd
oiwngEargoAn,andhe ofrnagment ocnisdmawinobre.ft12V.61S-0Cerppleerculosornolacyoiddcsiaeenddi5c.1 SliuniaolnSawnerdeaa.dTepsluidspeeaksjeecstisowneorfchelupeldesocloutniioncsaslndcara sadrds
-
APlmamsomnaiCuonmcePnetrrfaltvioornooicniPaonsota:tW:eaAngienEgRfatfseoFncotlthleowOirnag Gavage
DuPont15302
Dont15000AN:Pages "aTnhaelcyaclailbesatiaonndcarvdseowdatshgneneeraatledsbtaynedgarredseioen s0maFligiceu1s)i.ngDahtespfeoarkesrtesalrautiioofnoswmetrhee Comparedtothecalibrnioncurves 0 eval theconcentrationsofthe 24921. T((eeCsiVtf.sour=bmssitttaaynnodfamrudrdiiefvorixmaittiyiifnotnehaengve'h1Ji0vc0el)ecrwoifaaoshteiemvnnae)lfuscartsecedeabdcychacdoloncictlneagndoaliieoovngnelsh.oefActocehofeefdifucfpiilecnitcioaeftonetvfsruiptlieons avcceauabrifedeisisatohtnncieocfoathlne0t1s0b%tantchetshaonudagrhdouctrittehesolautiHoans.kellLabratcy for Ttohdeemteemainnreesuhletcoofntcheenctornacteinotnroaftitohenveestfsiucbtsitoannscepfioeshe(r1esp2ectfiovedsocshidnoglseivnegllsev.elwaswed ``eSitafbiicliatiyownasceuxpaelsteadtbhywbielnitvhee oer ecomepsilngofthehecworiespfonidiongfmsianbiinlgyCoencseantsr.ation
Te
PlAamsmmoanCiounmcePnetfrlauioornooicntPaonsotteW:eaAngiengERfatfsoeFnocltlhtoewing Or Gavage
Dupon15302
Dube 1A530 ue ANALYTICALRESULTS A. Chromatography ap24p9r21olvaild3f.ommitnhieHsPL2C0c.onluemcno8n3)refsoolthveedrpeeskgwiatnh .rReecptrisonataveo LCMSIMS Ccohnrtroomla.toTgirasmwsasrdeestheorwminniedF5guecsompvaris.onoTefstthudbislasnecnstwviaenrowitdheoacdiweithouhte 0kmeegl.
andar gaa th 0 g/m.controlconning ten andr,Refer 0Fires2a)
B. Recovery Samples coDentcaeinlterdastiotnofaH.2s49u2s1foofrheceov2e.r3ymsgam/plLesevae wuamss9i.z3%edofnomblae.TThheemmeesssseedd choenscctayiolnmoeftHho2d4p92e1rfomtedes7a.5tmfcyorlyeovveerwahse 1i02e.1c%oonfcoenrt al.Bigaoorsnthheiessdydin.,
C.UelformityofMitag ConcentrationandStabileSomples Annaflyoticaleofameixfuogmocdeesniinagsioon nvrsiccaoillocniedxo0n Ahuogr:o.o200e4swpedrnalrByaboyx ae ShowninAppTeaeInddSciaryxTabl,e 1 hbelfyolloswlineguslfeosuimsmasraimzpleintghdrayesofusfoa24r9i2f1oprmeipetrysifnconvceicfriartiaoinaond
[SRT. Rem am AVlaes CYS aNomn Utharaipnonntnis 50 eraorm ass o1o am
+ snBS . __ tsew le 3 wo b DeS oneteoerp erods.dRdoaei edmtioonp1.1bdooocomprreodflartSwer.viSthovnda.
Fades, {Teht suebssatasnfceowFa-s2a492h1esmrplgeds pcroenpacreerdianodncsll(e+ct1o6d.9o%ooAfngosmi:na9l,)2.0a0d8eiqnucaicsyemditxiedt(hCV JTeesstshuabst1a0n)csenwdassabbltednehedvehiiclheewhOmen/elldsahpousrs.torterpenesfor evs. E. Conclusion R`seusbusltatnsctervoamsthmiasleydpsiospoefttyh(eCH-Y2e49s21hdaonsin10g),sol1utionrsfgordhe setlsy in2di0c%atoetfsaomhineatle)sst.nd
bl underthcondosofthesy.Tetscbtocewsrofound he Om.empl.
En
APlmamsomnaiCuonmcePnetrrlavtoiroonocitnaPnooastte-:WeaAngiengEfRfeacst oFonltlhoewing Oral Gavage
DuPont-15302
Demis Paes
S"aTaabllee.Recoveryof o24g92i1 A2dd2ed1toDosingVehic=lea
BE a
Taomi Mess Nomioaal
TTRacor
ee145 i26 onio pe 5.
Ror etrcierdn hcts preepnintof pv.ey le,Tecig sve lesan
TulS1. arm ofMiigCommonSVearsnionasdSil o1001inDov= eSho
piam s Tel Vest dewsd
UsbCrni=nCommnion I
row
--
"-
i2ss
2m00
rLios
.
2s He:i1o"191a013
ass) uss
a
kA Mea 81e77v20a16 atListae)
SHA
23s5
i2n5
isas
Op ONr iib meecectrt srereiton ?foeen3.pSTteS,P10dof, cet
a ET ena open.
= --_--
A me CoT ncertoR in og aAngSRt Fallon Or Gorse
Dupanis30z
presi
re
nes
RepreAnsalvsiicaa Ctaliabrattioin Cvorvees
[ Vossen-- staat ox 10] Resour 09785
--
gi
i-
-
. TT Rw
ok
Connon,pm
FN o-- y--yha-- test_ SoroS-- ss
--_--
427
moni Pocnooe:cAagoBRlot nFolhlowingOr Gave
prs
Few
Representative LOMS/MSChromatographyChromatograms
Dumas
f Se e Ee
epeea
ae
he E= T e i i=ed o :ion mn
PAlmamsomnaiCuonmcePnetrrlastoiroonocitnaPnooastte-:WeaAngiengEfRfaetcst oFonltlhoewing Orsl Gavage Oo15502ANPugs Fi2gcuontroueed) `Representative LOMSMSChromatographyChromstograns
DuPont15302
Figo{RoeptreemotaanvdearL.ONMeSgaSivvoeraoinoPgFaOeAnrofec0t0rooile.csopueroilaomiinlo3f.y5Rm5i2s1.withe igo2r(R0e03p0rpstaivweithOeNSecrvumstiaondgarndo(f0F.0.321257p0e3-.26N3e(pAtPiFvOoAnaloyticPaltSFetaindnOaird)sA:
pony1.5 mic. --_
PAlamsmmoanCiounmcePnetrrfaltuicoronocintaPnoosatte-:WeaAngiengEfRfaeicst Foonltlhoewing Oral Gavage Dros1550 Pug 10 Figure coatioued) `RearsseptativeLOMS/MSChromatography Chronstograma
DuPont-15302
gar2e RCeporenmTnihenemeoefe0sL.CSs0o3epdcmmcroooinlmtieoopg4i9no2nofofermerggeip.rodneoiPnFgOssAllwsaiiilchnnislza$at6id8oFnoiAopeoL3u5t.]
-- Figaro Reb-- porreomonacioecLenErMiSionroofa0iogpanmooffHi2-- gh921coFveornydgoiangsonaPnFO(A7w4i5tohee.t)idcad
{ilmusng47p51nmig3..m5silniy,Tos siredcocofeFe
NT)
Tos
APlmamsomnaiCuonmcePnetrrfautoiroonocitnaPnooast-eW:eaAngiengEfRfaetts oFonltlhoewing Oral Gavage
DuPont-15302
Spee b Aapats,e 204 s col
Table
`SSonuar sofDopsineg niAossnes
Drieoi udsuie ofB42 (git)
[oY]
is
ow
ro
"
a2w0n
aisn
n
a1n0n
a"no
AvAvrergaegePMeerwceanrtNedoComnae
[I3
aaisn)
`SCtoanntdiacradtDoefrVitaotnitnt
wwon
oir
SshtoatyonTepes
2%
in
awn
sm
Temiad os aolch debeedondphee n.SwvFisvi1d)vi Br 3 rNeaumkmbeersiopersecdreeespieneeeotfsfikdd 3pres beddi,sdsof
- a.
PAlmamsmoariCounmcePnetrrfaltuioornoonctPanoosatteW:eaAngiengEfRfaetcst oFonltlhoewing Ora Gavage
DuPon-15302
Appendix B PFOA Plasma Concentration Da-tMalaes
Be
PAemiosriCmonPceaeorrnooncaPnocsW: eAgge ERla eoFoncllhtoewing Or Gove Awe TiDmoesPeost Aumsl Dose
GrTow (v3k) (ho2ws) _Nom1ber (m1p0h) 11 33 22 32 1100 {1 33 22 54 100 imw 33 22 150 El50 Im 33 22 n15 5500 mv 3s 32 316 5100 vv `` 22 55 1100 vv `` 22 s5t 1100 wwiiss 32 55s 3E0l] wwooooss 22 55 3300 wx is5 22 a5 3Io0 xx 55 >> w ww w xx 55 >> ww s wow xXti 55 22 75 El30
xxtt 55 2> w 5 x30 x 5 > wx
CPoFmOcAePnlvaasimoan G(g0m) "39330 S361595 1D5i6e2s 1105436 1S2a8 B35B85 i39s7s0 T$6.s41 1231175s3 1m24x03 25177050 32099 S08e1 57x3160 5
Dwron1si02 Maewm aSoDr mes 2% wera iA 0 wm 6H w% BH
Ex 33
APlmamsomnaiCuonmcePnetrrfaltuoiroonocitnaPnoosatt-e:WeaAnignegEfRfaetcst Foonltlhoewing Oral Gavage
Time Post Age Dose Animal
Dose
Growp T
(weeks) 3
(hours) 24
Number 9
(mg/kg) 10
1 1
3 3
2% 2%
13 1
10 10
1 1
3 3
2%
18
2 2%
10 10
nm m
3 3
2 2
7 30
19
30
ji m
3 3
2 2
2 2
30 30
m v
3
2 2%
2s
30 10
v v
4 4
26 26
10 10
v v
4 4
26 27
10 10
vir vit
4
2% 2
6 30 61 30
vit vit
4
2%
4 2%
8 7
30 30
vi X
+ 5
2 7"
30
2 108 10
x x
5 5
2 2%
109 10 10 10
x x
5 5
2 2
ns 116
10 10
Xi xi
5 5
2 2%
106 30
in
30
x1 Xt
5 5
2% 2%
in 13
30 30
xt
5
2%
4 30
PFOA Plasma Concentration
g/l) 3523 21.00 36.13 an 3649 6530 76.80 8005 98.90 973 1008 268 3 48.85 35.80 2.17 106.94 108.70 96.60 79.64 37.05 39.60 052 39.05 4680 9830 98.70 103.53 153.89 114.88
DuPon-15302 Mean SD un 189 716 1823 ast SB 0081 13.8 we 36 386 2536
-
The
APlmamsomnaiCuonmcePnetrrfaltoiroonociinaPnooast-eW:eaAngiengEfRfaetcst oFonltlhoewing Oral Gavage
DuPont15302
Appendix C PFOA Plasma Concentration Data - Females
ehe e
APmemmoaniCuomncPeenfrlauiooronoicnaPnosotes: -AgWe EeRcaattnFolnilhonewigg Oral Gavage Age "TiDmeesPcost Animal Dose
GroIwwp (we3eks) (ho2ws) N2um6ber (ma10le uu 33 22 37 1100 uu 33 22 3 1100 wv 33 22 3280 3300 ww 33 22 325 3300 wwr 3 22 376 3T0o vvii 44 22 77s% 1100 vvii 44 22 Eni] 1100 vviirn 4i 22 726 3300 vviinn 3 22 n5 3300 vxin 5s 22 1822 3010 xx 55 22 11246 1100 Xxx 55 22 1I2E8 1100
xIir 55 22 111187 3300 xxiu 55 22 112210 3300 Xi 5 2 1s 2
PCoFnOceAntPrlaatsimoan egnt) 445970 333826 9896.9654 074.4113 7M8e 311902s7 12919418 T619E8 7790.3185 w822e8 3238686 33405018 9T8A.6T1 33458565 3130
Dupont15302 M1e8m SSTD sass 1031 mE BIS
TT mm Tal S600 2946
---------------------- ETH-------------------- "%3C
APmamsoaniCuomncPeensrhaaioornooisnaPnoosseW:eanAignegERtfsoFnotltlhoewing Ors Gavage
DuPont15302
Grow
Age TiDmoesPeost Animal (weeks) (hows) Number
(mDogsse)
PCoFnOceAntPrlaatsimoan gnl) Men
SD
n1 33 2244 "0 1100 1538.720 35s 38
u1 33 22 16 1100 11933786
V u 33 a 2 w50 w 10 se12x0s8 sis nel
wwv
33 22% 32 3300
5105 22
r wwvv 33 22 a ss 33M000 3nlm 6e1is3 ies 0
vvii 44 224 88 110 2242004
vvii 44 22% 954 1100 m4 2+ 8 30
57.3607 30 mol 9%
vvinn 44 2244 358 3300 2391361
vviinn --X
44 224 sw
22 3500 m0 M0
21470392 2& 1% 08
xx 55 22 115321 1100
005326
r xx 55 224 113m5w37 s 11000 o2100s20 sa 195
xxii 55 22% 113554 3500
i55n3
xx1r s5 22 113386 3300
a23n0
eee ------------------ os------------------ +37
APlmamsomnaiCuonmcePnetrrfautoiroonociinaPnooast-eW:esAngiengEfRfaetcst Foonltlhoewing Oral Gavage
DuPont15302
Appendix D. Supplemental PFOA Plasma Concentrations
ie
A`PmlmasomraiCuonmcePnetrrfautoiroonocitnaPnoosatt-eW:eaAnignegEffReacttsoFnoltlhoewing Oral Gavage
DuPont15302
J Avimal PFOA Plasmga/CmoLn)centration Mean sp
Fasted Male | Fasted Male 2
3428
6495
234
prs
Fasted Femal|e Fasted Femal2e
5820
6816
ui
713
`NNoonnF-Faasstteedd MMaallee 34
281
3000
028
3020
NNoonn-FFaasstteedd FFeemmaalel4e3
310 2014
254
764
_ote: wAelreanciomlallesctwede2r4h3owuercskpsosoftdgoesinagn.d receiveda single 10 mpi dose. Plasma samples
-
Ts
APmlmaoinniCuomnPceernfvarooonctinaPnocssiteW:eaAngienEgffReacttsoFnoltlhoewing Oral Gavage
Dupont 15302
Appendix E Urine PFOA Concentration Data - Males
-
oe
440
APmimoniuomnPseerrlioroonocitnaPmoosstW:eAngge ERatcFoonlttlhoewingOrsGage
Duponw1sa0z
Animal
PCFonOceAntUrraitnioen
Grow Age(weeks) Number Dose(mgs) _ (ugml) Mem
sp
T0
33
15
1Io0
1962261 057 486
00
33 i1s 1100
134917
C In1 33 a11w7 3w500es e721.e.6990 S76 EE
IiI
33
221
5500
0N9s7
v m 3 25 50 e eanmds am as
vv
ss
6&
1100
356452
A vv 34 5n 0 1100 e i750e815: ma ima
vviit
3`
6a 3300
2105959
FES vvii 44 77 3300 E 3s5a4r7 EEE
xx 55 11009 1100
S2o3d5
a xx x55s 1111s 56 110 ta5281i50s Be em
xxii ss innz 3300
21536715
xxii 55 113s 300
1107.4166
NS =no rinesamplewas prodicedbytesnml
eereeee--=o----e-- i --r er-- ee +7
APlmamsomanCiounmcePnetrrfaltuiooronocintaPnoosatte-:WeAsgnienEg fRaftsoeFnocltlthoewing Oral Gavage
DuPon-15302
Appendix F Urine PFOA Concentration Data - Females
Ti
z
--
ADvmCoonntePnrtfiroocniainnPcoaWtiec,aAignegBRcas aFonltlhoewing Or Gove
Durencisi02
Animal
CPonFcOeAntrUartiinoen
Grosp Age (vesks) Number Dose(mg) (agin) Mean
SD
Tu
33 0ry 1i0o
11973596 20789
nu
33 iis I1n0
2101034
s n 3w 0 w 10 eSaswe
wwvv 33 re
00
59356875
~nv 33 i5s 55 rem nw
1s22e.s17 maB 58
ii swt 1100
271i769
N hwev i4 o0te 110 m3 1s7ss wim me
vviinm ii 5 300
3susse
vvmm 44 05 00 sew
1132161s6 am wm we
xx
55 1112 1100
$97865
s xx 55 w 11537 100 s2a8ws mie sis
xxuu 55 11334s 20
1sssa
xxuu
55 113s6
00
117754.2454
=
+43
+Ga= 4s