Document 935dZ88emRoL6qd5yM9nqO257
DuPon-15302
Study Title Ammonium Perfluorooctanoate: `Age Effect on the PFOA Plasma Concentration in Post-Weaning Rats Following Oral Gavage
TEST GUIDELINES: Noneapplicable AuTHOR: Paul M. Hinderliter, Ph.D.
STUDY COMPLETOEND: December 2, 2004 PERFORMING LABORATORY: EL du Pont de Nemours and Company
Haskell*TM Laboratory for Health and Environmental Sciences Elkton Road, P.0. Box 50 Newark, Delaware 19714-0050 LABORATORY PROJECTID: 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 3M Center Building St. Paul, MN 55144-1000
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APlmamsomnaiCaonncPeentfrlaitoiornooinctPnooss-tWe:aAngienEglRetcstoFnotlhloewing Ora Gavage
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GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
"Thisstudywas conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards, except for the item documented below. The item listed did not
impact the validityof the study.
+ No conduct audit was conducted on this study. however, all activities were performed by
experienced, trained personnel. All work was documented in the study records.
StudyDirector:
M. Hinderlter, PRD. Research Toxicologist
o3-eDtee200%
TT
PminrsiCoPneersesoinPnose-W:eaAtginegiResotFnotllhoewing Or Gevage
QUALITY ASSURANCE DOCUMENTATION
Dureneisicz
St`WuodrykCRoedqueesNtumNbuemrb:er: 113583939 dTahtescoonfdiuncstpeocftihoinsssrteuidnydhicaatbeedebnelsouwb,jected to periodic Quality Assurance inspetions. The
Phase Audied_ Audi Dates Protocol: July 2,23, 2004
ReportRecords: October 28,29, 2004
SDtautdeyRDeiproerctteodr to Niy23,2006
October 20,2004
MDaatneaRgeepmoretnetdto July23.2004
November 24, 2004
Reponsdby: T
C Tose C. Rl amil eor paee dey Quality Awsance dion
APlmaimnsoCnoPsnioeosanenPotse:WeaAtignegBRiacsnFtohlleowing Onl Gate
Duboarisi02
CERTIFICATION
We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study.
Awaits Evaluation by: ___ PotoaCiMMaldeaCchrematis5it5nr
oflDee zoodt
ET-- T =m6 w2 mmmp| 3b e-aivw -zoor Nees bane
esed by Study DireC cto: r FRINeGseacdhe TioritrologPi55.
02=De=c_zo0t.
-_ TM m-- m mm
PAlmamsomaniCuonmcePnetrrfaltuiooronociniaPnooastte-:WeaAngiengElReacttsoFnoltlhoewing Oral Gavage
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TABLE OF CONTENTS
Page
GOOD LABORATORY PRACTICE COMPLIANCE STATcoEc MENT
QUALITY ASSURANCE DOCUMENTATION..osvsssssssnessssmssmsmensssnenssnnnd
SA
----
|
STUDY [FORMATION coxmsaunsmmmmmmmmammmmnswmmssmsmmsdl
SONNY.onnsmmmmmmesnrmmmmmmmmmrmmrem--------r
A
"
'
TRIROVUCTION, ourgmmmisssisismmssmmmmmmmmmmmsell
MSTERIALS ANDMETHODSconmmmmmmmmminmmstmmsmmmmmmmenl
A. Test SUSIANCE rrr
m------------------
B. Test System A C.. Animal Husbandry verre
10 ----"
1. Housing...
tH
1
2. Cage and Cage Rack Change......
small
3. EOVIrONMNtal COMIONS rrr
1
FL --
--
5. Animal Health and Environmental Monitoring PrORam oor]
D. Pretest Period...
------
12
EL ASSIGNE0D GROUPS...
errr
Be MABSURY ovvrmsmsmmmnennen
-- ------
A
----------l
2. Dose Preparation, Analysis, and RALES 3. Sacrifice and Blood Sampling..............
ore
13
mmm
4. EXPETMCNIal PIOCCAU0. G. SUPPICTIEa SUA...
13 snd
H. Analytical Methods...............
i ------
1. Verification OfDOSE SOON... cere
14
2. Extraction of PFOA from PIasma Ft --
for
LC/MS ARGIYSIS........vovenrmroecrorercr wosmmm------------------
4
1. S4.HQSCIHGIEO)MBADOBGITYASPESIcoMvEtrhOUS....rrrrerrrrerewrrommmm-- ----_
I
J B. Plasma ConcentraoftPiFoOnA ...v.v.vovevrvrver
nnn 16 sane
2. Female Rats...
sns----------
16
C. U3.rinSe COoRncIeAntRrSaoSftPniFoOAnm..m. mmmmA -- -- SN"
a ----
RECORDS AND SAMPLE STORAGE vocvvvvrsrnrnrsrnrmrsmsmmsranensrsmmmsennens1r8n
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FL
------------
ET
--
Table 1: Plasma and urine concentrations in male rats ralgavage with 10 or
FL
---
Table 2: Plasma and urine concentrations in female rats oralgavage with 10 or
30 mg/kg PFOA
tran
2
ES
--------
Figure I: Plasma concentration in rats folloraolgwavaigenwigth PFOA _.................25
Figur2e: Urine concentration n ratsfollowing oral gavage with PFOA..............26
ALPE SOICES wrTAMA]
`Appendix A: Dosing Solution ARIS .................
29
Appendix B: Plasma CONCENtTation D-aMAItES a rover
Al
Appendix C: P1asma CONCENtTation Da-FtEmaales.........vovesrorarmnenevsneneos 44
Appendix D: Supplemental Plasma Concentrations
a7
AAppppeennddiiFExx:: UUrriinnee CCoonncceennttrraattiioonn DDa a--FMt etmaalaales.es...
4s9t
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STUDY INFORMATION
9th Collective Nomenclature: Octanoie acid, pentadecafiuoro-, ammonium salt
Synonyms/Codes: ++ . + + + +
AFCm-m1o4n3iFuLmUpOerRfAluDorBoroacntadnoFaltueorochemical Surfactant (3M Company, Specialty Materials) cs APFO Perfluorooctanoate, ammonium salt PFOA 2021 Lot 332 (3M Specialty Materials) (Lot No)
Haskell Number: 24921
CAS Registry Number: 3825-26-1
Purity: 95.2% 97.99% Straight chain: 77.6% Branched: 12.6% internal monomethyl (non-alpha) 9% isopropyl 02% tert-butyl 0.1% gem-dimethyl 0.1% alpha monomethyl
Known Impurities: Cs (CsF;CO; NH), 0.01%
CCo5((CCs4FF1s1CCOOys NNHH"L)),,00..0433%% C1(CaFiCO: NHL), 057% Co(CiFirCO: NHL), 0.16% Monohydro APFO, 0.09% Monounsaturated APFO, 0.72% Undefined (possibly) substituted perfluorocyclo 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 of instability was observed.
Study Initiated/Completed: July 8, 2004/(see report cover page) Experimental StartCompletion: August 9, 2004 /October 13 2004
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SUMMARY Perfluorooctanoate (PFOA) plasma concentrations were measured in immature (3, 4 o 5 weeks ofage) rats following single oral gavage doses at 10and 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, ae significantly different from the main study levels or both male and female ats. The plasma concentration variability in 4-week old rats suggests an unknown physiological variabiliy in animals younger than 5 weeks. As a result, the relationship between age and PFOA plasma concentration is undefined in 3-4 week-old rats following oral dosing with 10 or 30 mglkg PFOA.
A Plasma
PFOA plasma concentrations in malerats were not significantly different by sample time (at 2 and 24 hours) or by animal age (3.4 or 5 weeks)a either 10 or 30 mg/kg. except at 2 hours and S weeksofage. The PFOA plasma concentrations in female rats were significantly lower at 2co4ncHeonutrrsatpioosntsdowseirnega(lcsoomlpoawreerdftoo5r2 hweoeuks-)0latdarlaltsagtehsanan3dadnods4eswteeeskte-d0.ldFreamtaslaet bpoltahsmsaample times and doses (p< 0.01). The PFOA plasma concentrations following a 30 mg/kg dose were approximately 1.5 to 4 times higher than those observed following a 10 mg/kg dose.
Ina prior study, male and female rat PFOA plasma concentrations were measured at 24 hours
post-dosing following a single 10 mg/kg oral gavage dose. PFOAplasma concentrations are
consistent with the current study for bothsexesat 5 weeksofage but not at 4 weeksofage (3-
week old animals were not previously measured). The standard deviatariesomnalsl in both
studies, indicating that both are studies is whether the rats were
internally consistent. fasted before dosing.
The The
only known variation between the supplemental rats were added to
two
examine the effectof fasting on the PFOA plasma concentrations in 4-week old ras. Fasting
increased the PFOA plasma concentratioans expected but the nominal vales ae different from
tplhaesrmathceonccuernrternattisotnusd,y oitratphpeeaprrisotrhsattutdhye.reGiisvaennutnhekncoonwsnisptheynsciyoliongtihcealS-vawreieabkiloiltdyriant rPatFsOA
younger than $ weeks which impacts the pharmacokineticsofPFOA.
B. Urine There were significant differences in PFOA urine concentrations with age, dose. and sex. Female rats showed higher PFOA urine concentrations, and these increased with age. Male rats showed the opposite, with urine concentrations decreasing with age. Both sexes showed higher
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(2.56.5 times) urine concentration at 30 mg/kg compared fo 10 mk doses. No atiempt was. made to quantify urinary output (volume), and, because creatinine levels were not measured, adjustments for rine concentration could not be performed.
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INTRODUCTION Ammonium perfluorooctanoate (APFO) is a synthetic perfluorinated octanoic acid used as an iisndsuesxtr-idaelpesunrdfeancttanatn.d aEplpiemairnsattioobneoufntdheerdihsoarsmsoonciaalterdegaunliaotni,onp.erPflFuOorAooecltiamnionaatteio(nPiFsOmAu)chinfraasttser riantsf,eamnadleupra-trsegtuhlaanteind mbayleestrraasd,iodlowinn-trheegmualalteerdbaytste.stAostperrioonresitnudbyotchofnedmuaclteedanadt DcausPtornatted male Haskell Laboratory found sex-dependent PFOA elimination in 4-8 week old rats") The sex "dTihfefeorbejneccetiwvaeosflteahset atthi4s wsteuedkysowafsagteo,daettetrimmienpeierfio3d-wjuesetkproilodrmtaolseexaunadlfmeamtaulreatriatosneilnimtihenartate. PFOA from blood atarate different than that observed in 4- or 5-weck old rats. Male and female pweorsetwceoamnpianrgedratast a2tadnidff2er4enhtouargsespowsetr-edodsoinsge.d wAitth24APhoFuOr,s aafntderthdeosPinFgO,Athpelsaesxmadicfofnecreenntcreaitnions oPfFPOFAOcAl.i)minUartiinone iwnassexcoulallelcytmeadtfuorre2r4athsouwrassaenadsialnyadliyszteidngfuoirshPeFdOvAiactohnecepnltarsatmiaonc.oncTehnitsrasttiuodny was designed to evaluate limited Kinetic endpoints exclusiveofdeterminationof limination rate constants.
MATERIALS AND METHODS
A. Test Substance
AupPoFnOrewceaispt.obtAavianieldabflreomin3foMrm(aStt.ioPnauoln, tMhNe)puarintdy,acsosimgpnoesditHiaosnk,elclonLtaabmoirnaatnotrsy,NsuynmobneyrmsH,-24921
hazards, the study
and hazardous material records and/or report.
classifications)
were
provided
by
the
vendor
and
documented
in
B. Test System RMaalleeigahn,dNofremtahlCeaCrorllin:aC.D(ASnDim)aIlGsSfBoRr grartosupwsereIVobtaaririnveeddfarsolmitCtherasrwliesthRidvaemrs.LaTbohreatSoprriaegs,ucI-nc. sDuaiwtalbeilyitryatwwitahsrcehsopsecetntfoorlotnhgisevsittuyd,ysebnesciatiuvsietoy,ftahned elxotwenisnicviedeenxcpeeorfiesnpcoentwaitnhcotuhsisdsitsreaaisnesa.nd ts Faunrdtohtehremrorfel,uotrhienaStperdatgesutem-aDtaewrliealysrat has been used previously for toxicokinetic estingofPFOA dAtithed nottiemxecoeefddos2i0n%g,ofrtahsewemreeaanpwperiogxhitmabtyeldyos3e, 4g,roaunpd a5nwdeseekx.soIfnafogremaantdiotnheonwetihgehctagvaerilaabteilosn. "iTnhceluidneddivtihdeuaHlasikdeelntlifainciamtaiolnnnuummbbeerrawnadsapnliancdeidviodnuatlheidteanltoifficeaatcihonranubmybetrataososiogrnteaidltmoarcka.ch at.
Tho
APlmamsomnaiCuonmcePnetrrfaltuiooronocintaPnooastie-:WeAagnengEfRfatcstoFnoltlhoewing Ora Gavage
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C. Animal Husbandry
I Housing
Upon arrival at Haskell Laboratory, rats were removed from shipping cartons and housed in arepcporrodpsr.iaRtaetcsagweesreacmcaoirndtianignetod SutnadnedrarqudaOrpaenrtaitneifnogrPartocleeadsutrethsreuenldeasyssspuencliefsisedapipnrtohveedstoutdhyerwise by the site veterinarian, had at least one recorded weight gain, and no abnormalities detceted. After the quarantine period, rats were selected for study.
During pretest, dams were housed with their liters in polycarbonate pens containing bedding.
Throughout appropriate
the dosing period with cages according to the
test substance, the rats SOP. During the urine
were housed individually in collection period, rats were
housed
in
stainless steel wire mesh cages.
2. Cageand Cage Rack Change
Due to the lengthofthe study, no cage and cage rack changes were required.
3. Environmental Conditions
hAunmiimdailtryooofms30w-e7r0e%m(atairngteatiendetdoa4t0a-6t0e%mp)e.raAtnuirmeaolfr1o8o-m26swCe(rtearagrettifeidcitaoll2y2i-l2l4umCi)naatnedd arelative:
(diuroercteosrceorntthleiglhatb)oorantaonryavpeptreorxiinamraitaen t102-hhaovuersliiggnhitfiidcaarnktlcyycalfef.ecUtendletshsejruesdugletdsobfyththeessttuuddyy, the
lative humidity and temperature included in the final report
ranges
in
the
housing
rooms
were
recorded
but
were
not
4. Feedand Water
ARlolderantts LwaebrDeiperto"v5id0e0d2atdapliwbaitteurm.adRlaitbsitwuemraendnoftefdaPstMeIdprNiuotriotidoonsiInntgerdnuaetitoonatlh,eLiLaCge.Certified
5. Animal Health and Environmental Monitoring Program
AfoslslpoewciinfgiepdroicnetdhuereHsaswkeerlel pLearbforoartmoerdypaenriiomdailcahlelaylttho aennsduernevtihraotncmoennttaamlinmaonnittloervienlgs pwreorgerbaem,lotwhe those that would be expected to impact the scientific integrityof the study:
+
Water samples were analyzed and other contaminants.
for
total
bacterial
counts,
and
the
presence
ofcoliforms,
lead.
+ sSaanmiptalteisonfrboymthferecsahgleywwaasshheresd.cages and cage racks were analyzed to ensure adequate
rCeerqtuiifrieemdeanntismaanldfneoetd twoaesxucseeedd,sgtuaatreadnmteaexdibmyutmhecomnacneunftarcattuironesrotfokmeeyetcosnpteacimfiineadntnsu,triitnicolnuadling
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pspreecsiefniceed ohfetahveysemectoanlts,amaifnlaatnotxsinb,eclholwortihneatmeadxhiymdruomcacrobnocnesn,traantdioonrgstaantoepdhobsyphtahetemsa.nuTfahceturer were not expected to impact the integrityof the study.
"The animal health and environmental monitoring program was administered by the attending
Iaboratory animal veterinarian. Evaluationof affected the validity ofthe study.
these
data
did
not
indicate
any
conditions
that
D. Pretest Period
Upon arrival at Haskell Laboratory, all ras were housed in quarantine. The rats were:
+ quarantined for at least 3 days.
+ identified temporarily by cage identification.
+ weighed at last 3 times during quarantine and once prior o initiationofexposures.
+ observed with respect to weight gain and any gross siogfdinscasse or injury.
The rats on body
were released from quarantine weights and clinical signs.
by
the
laboratory
animal
veterinarian
or
designee
based
Rwaittshotuhtatnewcerroepsayc.cidReanttsaltlhaytkiwlelreed ofrournedmodevaeddofrrwoemrsetusdaycrdiufriciendg itnheepxrterteesmt pdeurriiondgwtehreeprdeitsecsatrded ipenrcilouddewdeirnetghievfeinnaalgrreopsosrtexunalmeisnsatcioonnsitdoercehdecskigfnoirfitchaentprtoestehneceeovaflduiasteiaosneo.ftTehesrteusduyl.ts were not
E. Assignment to Groups
Rais were selected for use clinical signsofdisease or
on study based on adequate body injury. They were distributed by
weight gain and freedom computerized, stratified
from
any
srtaantdisotmiiczalaltyisoingniinftiocasnttuddyifgfreoruepnsceassadmeosniggnagtreodupinbtohdeySwteuidgyhDtemseiagnns,.soRtahtast ftohrergerowueprse InoV were
aelxsco ereanddo2m0i%zoefd btyhedmaematnoweenisguhtr.e eTvhene dfiisrsttri5burtaitoni.n Teahcehwgerioguhpt wvaerrieatdieosniogfnsaetleedcftoerdsaractrsifdiicde naott
the 2 hours and the remaining raisin eachgroup were designated for sacrifice at 24 hours.
Eidaecnhtifraitcawtaiosnansusmibgenerd. aTuhneiqlauset63-ddiiggiittsHoafstkhelelHaansikmealll nanuimmbaelrnaunmdbaenr iwnadsivtiadtutaoloecdagoen the tail of
cach cage
rat. The label.
Haskell
animal
number
and
cage
identification
number
were
both
included
on
the
fRoarisotthheart lwaebroerantootryaspsuirgpnoesdestooar steasctrigfriocuepd,biyncclaurdbionngdtihoexdidaemsasfprhyoxmigartioounpsanLdIVd,iswcearrederdelweiatsheodut anatomic pathology evaluation, at the discretionofthe study director.
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F. Main Study
1 Animals
"The experiments were composedofthe following animal groups:
MaleGrowFemsle _D(ngohg sWNedusAsmebDaeysroMarleRaFnemale
mt ow1 500 35 a aw w 01 vv ivvi 1 43 F3 T)ow 10 ixx xxn 3010 55 5300001m0 100
Andiomsailnsgwwaes droce1umdeanytefdroinmhtheesgtuedytraecgoradsthaGndlnor offldosreipnogrt.. The actal age at *"SG/isveex/ngsroaupsisnagclreifgiacvedagae 2doasned 24 hours potedose for blood collection
2. Dose Preparation, Analysis, and Rates
iTthse tthesetrsouubtsetmaoncsetwcaosmmadomnilnyisutseerdedfaosr taoxsiicnigtlyesotruadliegsavoafgAePdFoOs.e. ATPhiFsOrowuatse wmaisxecdhowsietnh bNeAcNauOs-e pbuordeywweiaghtttoawceehrieeruvseedthweittahrgaetdodsoesevoclonucmeenotrfaatpipornso.xiDmoatseellyev4elmsLo/fkg10baonddy 3we0imghgt.APFOTkg
Dcoonsdeitwioanss.prepared on the day of use or rior to the day of use and stored under appropriate
HPLC-MS methods were used to confirm PFOA concentration in the dose solutions.
3. Sacrifice and Blood Sampling
Rats were puncture.
sacrificed
by
CO;
asphyxiation
and
blood
collected
from
the
vena
cava
or
by
cardiac
4. Experimental Procedure
R`aTtwso wheoruresoabfttaeirndeodsifnrgo,mSthreatvsesnedxo/rgraonudpaedrmienissatcerrifeidcePdFaOnAd wathodloeseblleovoedlscodlelsecctreidb.edWahboovlee.
blood samples were placed into EDTA by centrifugation and frozen at <-10C
tubes and maintained until analysis.
on
wet
ice.
Plasma
was
separated
"cTohlelercetmedaifnoirn2g4hroautrss/sacfxt/ergrdoouspinwge.reAptl2a4cehdouirnsm,eatlalbroelmiasimnicnaggersatfsowreurreinseaccroillfeiccteidona.ndUrwihnoelewas blood collected. Blood was handled and separated as above.
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G. Supplemental Study
Four additional male and four additional female rats were added to groups V and VI. Two of the
male with
and twoofthe test substance.
female rats were fasted overnight for Dose preparation and administration
approximately 12 hours before dosing was the same as in the main study. Al
ats were sacrificed at 24 hours post dosing and whole blood collected. Blood was handled and
separated as above. HPLC-MS method was used to determine plasma concentrations of PFOA.
H. Analytical Methods I. VerificationofDose Solution
Methods for verificationofdose solutionaredetailed in Appendix A. 2. Extraction of PFOA from Plasma for LC/MS Analysis
Pprleacsimpiatastaimopnlceoslwuemrnes p(rJoocneesssCehdrboymaptrootgerianpphrye,ciLpaitkaetwioonod(,PPCTO)).usiAng0.I5soplgut/emALrrsaoyluptriootncoinf panerifnltueorrnoalnosntaannodiacrdacfiodr (quAalndtriitcahtiCohnemoifcPaFlsO,A.MilPwlaauskmeae,saWmIpl)eisnwaecerteontihtarwileed,(AanCdN)a 2w0asuL.usaeldiqausot oafpepraocphrisaatmepdlieluwtiaosnarpaptelivcodltuomtehseoPfPATCNar/riayn.teTrhael psltaansdmaardsasmoplulteisonwetroethperePcPipTitaartready.byDaidlduitinogn rsattaensd,arradnsgoilnugtiforn)o,mw1e:r1e0 (ut1i8l0izLed ionfoirndteerrtnoalcsatpatnudraerdthseosluatmipolne) tcoon1c:e2n0t0ra(t3i9o8n0s witohfinitnhteersntalandard cceunrtvreifpuagreadmeatter~s3.00T0herpamrrfaoyrwamsisnluotwels,yealnudteedxtruancdtesrwvearceuaunmaliynzteodab9y6-LwCe/lMlSr.eceiver plate,
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3. Instrumentation
System: DMeotedcet:or: SourcIe:M 1 resolution:
HToMn enerregsyoluI:tion EConltlriasnicone:: ELxMit,2 resolution: ToHnMen2rcersgoylu2t:ion: CMaupliilpllaircyr((kVV)):: CEoxnieac(oV)r: (V): RSoFurlcenest(eVm)p:erature: MS MDeetshoolda:tion temperatures MToimdee: EH
cn
hWeaatteerr,sa2n7d95atLoisquumipdlCehrromatograph, equipped with quaternary pump, column QMuRatMtro Micro Mass Spectrometer N1e0g0ative Electrospray 11000 s2 5mn 2u0 62500 i1s0 0noc 350C M0-R1M0,02mitnransitions Du4e1l3.l0:0 369.00 025 sec 4Co6l3li.s0io0ne4n1e9rg.y0:0~~ 15V DuCoellli:sion energy: 0s2e5vsec
4. Chromatographic Methods
MCeotlhuomdn: CMoobliulmenphtaemspeesr:ature:
PWlaatsemras cXtoenrcreantMraStiCo1n8a,naly2si.s 1x302.m5 mum, AA:mb5i0enmtM Ammonium Acetate B: Accioniile
Gradient:
[mem Twn] eo--Ts
C7 o 0 s
Ceo e10e0 0
SFtloopwtritmee:: Injection volume:
01205mmiLnmin Sil.
Ts
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I Statistical Analyses
Group data is represented as Mean by the Student's test
SD. Where appropriate, statistical significance was assessed
RESULTS AND DISCUSSION
A. Dose Solution Analysis (Appendix A) `dTehsecrciobnecdenitnrMaattieornisoalfsthaendteMsettshuobdsst,anSceectiinotnheF.doSsteansdoalurtdisonfosrwdeorseevveerriiffiiecdatbiyonLaCn/dMtShemdeotsheod as sthoeludtoisonesswoelurteiopnrsepwaerreedabllywdiitfhfierne2nt0p%eorsfotnhneelt.arTgheteeddectoenrcmeinntreadtmioena(n2.P5FaOndA 7c.o5ncmegnt/rmat.ions of Appendix A). PFOA was stable in the vehicle for at least 5 hours.
B. Plasma Concentration of PFOA
(Tables 1-2, Figure 1, Appendices B-D)
Lo Male Rais
`2T4hehoPuFrsO)Aorpblyasamnaicmoanlcaegnter(a3t,i4onos w5erweeenkosts)igatniefiitchaenrt1l0y doirff3e0remngt/kbgys.aemxpcelpettaitme2 h(aotur2saanndd
s5owmeeekgsrooufpasgtehe(p2<40-.h0o1urbapsoesdt doonstihnegrceosnucltesnotfrautnipoanisrweedrtetehsitghaetr,99bu%t cthoinsfiwdaesncneotinutneervxapl)e.cteIdn
due o the long concentrations
plasma ranged
eliminationofPFOA in male rats.) from 21.00 10 48.85 g/mL for the 10
Individual PFOA plasma mg/kg dose and from 49.51
to
w1e5r3e.8a9pgpr/omxiLmaftoerltyh2e-330tmigme/skghidgohseer.thPaFnOtAhospelaosbmsaercvoendcefnotlrlaotwiionngsafo1l0lomwgi/nkggad3os0c.mg/kg dose
2 Female Ras
2PFhoOuArsp)liansamlal acgoencseanntdradtoisoenss wteesrteed.sigPniFfOicAanptllyaslmoawceornacten2t4rahtoiuornsspowsetredoaslisnogl(ocwoemrpaatr5edwetoeks tfhoalnloawti3ngoar430wmegk/skgatdboostehwsearmepalpeptriomxeismaatnedlydo1s.e5st(op4<ti0.m0e1s).higPhFeOr Athapnlatshmoasecoonbcseenrtvreadtions following a 10 mg/kg dose.
Ee
PAlsminmaoConncePnetrrfautoironocitnaPnoosstteW:eaAngiengERfaitcstoFnoltlhoewing Ors Govage
Dupo 15502
3. Supplemental Rats
Individual fasted and
PnoFnO-fAusptledasamnaimcaolnsc,ernetsrpaetctiiovneliyn.
thPeFmOalAepaltassmwaerceon6c4e.n9t5raatnidon3s0n.00thpegf/emmLa.lfeoratthsewere
68.16 and 26.54 ug/L forthe fasted andnon-fasted animals, respectively.
Ipnoast-pdrioosrinsgtufdoyl.lo"wmiangleaasnindglfeem1a0lemgra/tkPgForOaAl gpalvaasgmeadcoosnec.enPtrFaOtiAonpslwaesrmea cmoenacseunrterdataito2ns4ahrocurs
`cowneseikstoelndtawniitmhatlshewecurrerennot sptruedvyiofuosrlbyomtehasseuxreesd)a.t 5Twheesktsaonfdaradgedebvuitatniootnasta4rewesmeaklsloifnabgoeth(3-
ssttuuddiieess,iisnwdhiecatthienrg tthhatrabtostwhearree ifnatsetrendablleyfocroensdiostseinntg.. TThhee osnulpyplkenmoenwtnalvarraitastiwoenrebeatdwdeeednttohe two
eixncarmeianseedthteheefPfFecOtAofpflaasstmiangcoonnctehnetrPaFtiOoAnspalsaesxmpaecctonecdebnuttratthieonnsomiinn4a-lvevaclkesoladreradtisferFeansttifngrom
ither the current study plasma concentrations,
iotratphpeeaprrsiothsattudtyh.erGiisvaenn
utnhekncoonwsnisptheynsciyoliongtihcealS-vawreieakbiloiltdy
riant
aPtFsOA
younger than 5 weeks which impacts the pharmacokinetics of PFOA.
C. Urine Concentration of PFOA
(Tables 1-2, Figure 2, Appendices E-F)
hPaFdOhAiguhreirnePFcoOnAcenutrrianteicoonnscdeinftfrearteidonssigtnhiafincamnatlleyswainthd atghee,fdeomsaeleanPdFsOeAx (upri<ne0.c0o1n).cenFteramtailoensrats
aignec.reaBsoetdhwsietxhesageh.adMhailgeherrat(s2s5htoow6e.d5 tthiemeosp)poPsFitOeAwiutrhinuericnoencceonntcreanttiroantsioants3d0emcgre/aksgincgomwiptahred
1010 mg/kg doses. creatinine levels
No
attempt
was
made
o
quanify
urinary
output
(volume)
or
standardize
for
CONCLUSIONS oPfearfgleu)orroatosctfaonlolaotwein(gPsFiOnAgl)epolraalsmgaavcaognecednotsreastiaotns10wearned m3e0amsgu/rkegd.inPilamsmmaatucroenc(e3n,t4raotrionwseveakrised sainginmiaflisc.antIlnytbheetcuwrernenetxspteurdiym,emnatsl,eersaptePciFaOllAy wplhaesnmacocmopncaernetdra1tioanpsriaorre srteuladtyivweiltyhcdo-nswteaenkt oilnd dmeaclreearastesdawcirtohssiangcreegarsoiunpgsa,gev,arbyuitnwgaosnloynlwyitshtadtiossteiclalelvyelsigFneifmiaclanetlryataptltahsem3a0cmongc/ekngtrdaotsieonlsevel. uPrFinOaAryceoxnccreenttiroantiionnfseimnalureinrea,swahtiallel nagoets.norTmhaelipzleadsmfoarcootnaclenetlriamtiinoantsiofnr,oimndtihceasteudppmloermeenrtaaplid gatrso.up,Thheowpelvaesrm,aacroensciegnntirfaitciaontnlvyadriiafbeirleintty ifnr4om-wtehcekmaolidnrsattusdsyuglgeevesltssfaon ubontkhnmoawlnepahnysdiofleomgailceal pvalraisabmialictoyncinenatnriamtailosn yisouunngdeefritnheadni5n w3e-4ekwse.ckA-soalrdersautsltfotlhleowrienlagtoiornashdiopsbientgwweietnh a1g0eoarnd PFOA 30 mykg PFOA.
7
PAlmamsomaniCuonmcePnetrrfaltoiroonocitnanPoosstie-:WeaAngiengEfRfatcstoFnoltlhoewing Oral Gavage
DuPon15302
RECORDS AND SAMPLE STORAGE
A sample of the test substance will be collected for archive purposes and retained at Haskell Laboratory, Newark, Delaware. Specimens(ifapplicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at ron Mountain Records Management, Wilmington, Delaware.
EE
PAlmamsomaniCuonmcePnetrrfaltuiooronociniaPnooastte-:WeaAngiengElReactts Foonltlhoewing Ora Gavage
Dupon15302
REFERENCES
1. Ohmori K., Kudo, H., Katayama, K., and Kawashima, Y. (2003). Comparison of the toxicokineties between perfluorocarboxylic acids with different carbon chain length. Toxicology 184, 135-140.
2. Vanden Heuvel, J.P, Davis, J.W., Sommers, R., and Peterson, RE. (1992) Renal excretion o7f.p3e1r3fl6u.orooctanoic acid in male rats: inhibitory effectof testosterone. J. Biochem. Toxicol
3. `KCuodmop,arN.i,soSnuzoufktih,eEe.l,iKmaitnaaktiuorna,beMt.,weOehnmoperrif,luK.o,rNinoastheidrfoa,ttyR.a,caidnsdwKiatwhadsihffiemrae,ntYc.ar(b2o0n01c)hain length in ras. Chem. Biol. Interact. 134, 203-216.
4. linen, M., Hanhijarvi, H., Jaakonaho, J. and Peura, P. (1989) Stimulation by oestradiol of
the urinary 27.
excretionofperfluoroostanoic
acid
in
the
male rat.
Pharmacol.
Toxicol.
65.
274-
5. DuPont Haskell Laboratory (2003). Ammonium Perfluorooctanoate: Age Effect on the
Plasma Concentration DuPont-13267.
in
Post-Weaning
Rats
Following
Oral
Gavage.
Unpublished
report,
6. DuPont Haskell Laboratory (2003). Perfluorooctanoie Acid: Toxicokinetics in the Rat. Unpublished report, DuPont-7473.
7. DuPont Haskell Laboratory. (1997). Hazard Characterization for Human Health C8 Exposure. Unpublished report.
8. dViasntrdiebnutiHoenu,vmeel,taJb.Po.lisKmus,laiknids,elBi.mIi,naVtainonoRfafpeerlfglhueomr,oMo.eJt.a,naonicd aPceitderisnonm,alRe.Ea.n(d19f9e1m)a.leTriatsss.ue J. Biochem. Toxicol. 6, 83-92.
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PAlmamsomaniCuonmcePnetrrfaltoiroonocitnanPoosstie-:WesAngiengEfRfaetcst oFonltlhoewing Oral Gavage
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TABLES
=
APlmamsomnaiCuonmcePnetrrfaltviooronocintaPnoosstte-:WesAngienEgffReacttsoFnoltlhoewing Oral Gavage
TABLES
EXPLANATORY NOTES
ABBREVIATIONS:
sD
standard deviation
Dupont15302
- a
APlmamsomnaiCuonmcePnetfrlautoiroonocinsnPoosstieW:eaAngiengEfRfaetcst oFonltlhoewing Orsl Gavage
DuPont 15302
Table: Plasma and urine PFOA concentrations in male rats dosed via oral gavage with 10 or 30 mg/kg PFOA
Age Dose Group _ (weeks) _ (my
v' hl5 1100 xs0 vmit 33 3300 x1 5 30
Data expressed ss gn. _2iloursPostDoPsleasma24 Hours PostDose Mean sp Mean sb
a99m2 aados 632 6%
u2n9
7m sx
we 360
W206As 11287180 e566 ISS
1T0a8le1
BB Bas
Is a6
Urine 24 Tiours Post-Dose Mean SD
957 ase as 2s as 236
SI% 2870
2886 Iss
Is6s 624
-
PAlmamsomaniCuomnPcefelurornoctinamPoossteW:earAigengERaftts Foonltlhoewing Ors Govage
Dupo1s30
Table2i
Plasma with 10
aonrd30urmign/ekPgFPOFAOcAoncentrations
in
female
rats
dosed
via
oral
gavage
Group (Awseeek) mDgohsee) v: 33 11 x51 wwiods 5o0w Xs ow
Dats expressed as gm. 3liaursPoscusPelasma24 Hour PostDose Mem SD Mean SD
woos wsims B23 a
mmesss s70m1 Ue om
Wsea 104sd se mes
MSOsL B9%e sa 19s
24 HourrsinPoescDuse Mem sD wma2r 1se5sa wm dies 1w0m2 297 i Suse
-
APlmamsomnaiCuonmcePnetrrfaltoiroonocitnsnPoosstWee:anAigneg ERfftectsonFotlhleowing Orsl Gavege
DuPont-15302
FIGURES
ETS
PAlmamsemaniCuonmcePnetfrlautoiroonosiinaPnoosstte-:WeaAngiengEfRfaetcst Foonltlhoewing Ora Gavage
Duont15302
Figure 1: Plasma PFOA concentration in rats following oral gavage with PFOA
wr,
"
ZF =
LS
=
3. EHoE.
:
2
=
| hh:
7
| [3C2342 hhoouurrss ppoosstt ddoossiinngg
I
7 lk7 L T
Age (weeks)
ab MMaallee ppllaassmmaa aatt 3100 mmgg//kkgg ((ggrroouuppss II,l,VVaInIdaInXd).XI).
d
Female Female
plasma plasma
at at
10 30
mg/kg meg
(groups (groups
I, V1 and X). IV, VIII and XII).
-_--
AimimmonCiosnPeraroocanPnositeW,aAigenERlosoFnoltlhoewing Or Govage
fr
Figare2: Urine PFOA concentration in rats followingoral gavage with PFOA
1s
|
a
b
150
gE
S$5 10 7s
8g g
5 = Loe
Pies IRI 3i
male BEX] female
EIN NG oBts
mr B aBs Y [otets!|
B ge OR OR
RY BY
BBlY OB B KBY
INGE
Be] NE BR
INN ENN NN
5
3
i
:
Age (weeks)
a3010 mmge/kkg ((ggrroouuppss II,,IL,IVV,,VVIIL,, IVXIaI,ndXIX)a.nd XID,
I
PAlmamsomnaiCuomncePnetrrfaltuioroonociniaPnocast-eW:eaAngiengEfRfeacstFoonlltohweing Oral Gavage
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APPENDICES
-
PlAamsmmoaniCounmcePnetrrfaltiuoonroocintaPnooastte-:WeAagniengEfRfatcstoFnoltlhoewing Oral Gavage
APPENDICES
EXPLANATORY NOTES
ABBREVIATIONS:
L0Q
limitof quantitation
sD
standard deviation
SE
standard error
%Cv
percent coefficientofvariation
DuPon-15302
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APlmamsomnaiCuonmcePnetrrfaltuiooronociniaPnooastte-:WeaAngiengEfRfaetcst oFonltlhoewing Oral Gavage
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Appendix A Dosing Solution Analysis
EE
APlmamsomnaiCuonmcePnetfrlautoiroonocintaPnoostte-:WeaAngiengEfRlaecttFoonltlhoewing Oral Gavage
Dupont 15302
AmmoniumPefluoroo`ciasWosieeR:aAaigsFenoElfleiocwtionnngOhrgealPGaavsaageConccatratiofon Post
ANALYSIS FORH:24921INDOSING SOLUTIONS
MeHdaiskceallSRaemspelaecNhuPrmobjeerct:Number Ansys] Report Number:
2153139 DuPont 15302en
SUMMARY. c`oDlooscitngesdofaoioanisforcmoinyceonftmratiionxsofi2n, g7/.v5mcegr/foimc.ntioofcnan2rd4s9t2b1irweraenpiaryeospa(n5rsedd. hsoorurobomfmeoraimnatpl eewoidrtnhAatuhegaucstrotf,se2a0m0p)4l.esI.addin0mg/mL. (conv)samplewas iCuoinccehnrroafioo24mnt9a2sn1adienmsmtepsarsoatsepdegocstirnormgestlrayui(opLCn-sMwhSeIrMySm)e.asuredby high performance Thesuspensionvehiclefo hestywasNanopore water. "Thhee rsetssuubsstoarnHc-e2w4a9s21astameptlaersgeptreedpcaorendacedrconllse(ctxed1o6n9%Auogafsno9m, 2i0)0,4ianddeiqcutaetdeltyha mfioxraeldl e(vCVsJ.essthan 10)andsablef thevehiclewhenheld hours 3 oomtemperatre
H25425wasnt tectedinthe 0mn. (onl)samples.
--0-
APlmamsomnaiCuomncPeenfrlauioornooinsaPnoosa-eW:eaAngiengEfRfetcst oFonltlhoewing Ora Gavage
Dupo 15302
Deron per SAMPLE SUBMITTAL cDooolosleimcnc tgesdofoie rnwnsior rocmsotymAeuoa fpnmitio3in,) ns0go0cfo.2n51,caebs7oon.nvm,enOgtL.ioo.nf 1(52c4so9a2nb1)iswepsetlpryewpe(hbsoeodsrted orn wilh at of pes. `Supisewsausbmditcedtfo.relswer ralyed eda eywer ceivedanoswhen
Thessensionvicerhe dy wasNeopur wee:
METHODS 1. AMvs ens + ResorSampleAnlsia (CoNceaneiwhedeviwseseiodnstseaoywsvel (s0.5mopl2)4a9n2d1 srohmshkevis1.e3mind) c ipr,oxo Fiomraahlella sc1.yo2incemisnealGomonwr e).edea.lnsd,A7s05oscmpkgrso(lpuegishoi)neofotfuhoeAot9fm2h1eswrasisi]tiwcoapnsud0eoidncN0is0op0arie.of Niomonpvuerheiclwe.aTroAslalmpcloevsweeyesGpeinepsrwoecesehdeanndianensyora peoessoeofaorenst 5ei Goingsane smsconcions. b. DosingSuspemionTrsmens ENxicoohpuidroesetinggvs1ia2em4rp9l02e1(0in5exmhpLee)cawtsaedpsceiontne.cdTehr0end1o0os0fimsnlgcawsmitpihlaeNaslnwoypeoe0re0t0w0ea0rtmelrgai/lde.dipwsicetrhd0 Sodsey)wBseaeedlfchdusoss,ihemgeesorkavlnhflOmcgoncesrunp,eof(ri eel. Santosets nl pis. Sahmepnlresbeiwd fsonsns l.swereany he ay he lionswerrcvadfox
Si
PAlmamsomaniCuonmcePnetrrfaltuiooronocintaPnooastie-:WeaAngiengEfRfaetcst oFonltlhoewing Oral Gavage
DuPont-15302
Doe1530 A Pages . CumsiogaphicConditions
CLoatlrseus:aintes ZootraAORlCSi7.925c.H1PmeLmC.x 150m,
Flow Rae:
smmimi
CIonjlecutimoTn Veomlpa:ers: 325pC
CMoobliolerPShwaistec:h: 50015m%iAtcercActicin
GrTeiente50(nia) % Asckonicle
0i5o
i0
55801s
5s
TMrSouenmeeters
MiQcuroo MincoTasndemsMS
Coonpileoynvlcaeg:e E 2ic (ES)soegatn veon
SoCounesVoTtaege:ngen: s120v
SDeeanvatnioTn:ope:P31F5O02A0: C41P3FnOiA:pr4e15t)ls03(6p0arei)e0G.n37g0hris)(Bagh)
RetinTir:of PFOAan 1.245'C PRON:appe3x.3miinzesm20asefelctiyon)
4 CaibsrdQauniitaotionn
Aacsettocoknseol.utiAopnproofpAeviasleyatlciaqluoSttoanftdaerds(cAePFwOe,reH-i2t2e70d3.w2i6t5h,N0a0n%appaguwe)rawearstnaadkeein Bceafloibrreattihoensseaanlidqaurodtwstehratbbarocukghetdtohveoalurmge,scaonpcrricoatpironeosfmohcendsiofedmesaplltsndlauctds(.12 C perooroctnoacid)was added. ASIpnenctarFomlEeotfGyrHyA.s(2Le4iC5-2sMs1SwIaMssSb).ylbNicegdpgewrafooimtnalnicecteovleipeqSruia.diychnisruocmeeadtto polegeaorpshrynesfgnamtddoaeeslmiyscdhard {nitrnngaarlgoan,nada.Tfrriapghmceantteiejnecstmioonniotforeds.p1l.261-sColutipones nrdfcbsbeosotioonicdtdairnedsdoi1c Solutionsweremaceanpeak areaswerecalcdcectroniclly.
Tas
APlmamsomnaiCuonmcePnetrrfaltoiroonociinanPoosstie-:WeaAngineg ERfaftecst Foonltlhoewing Oral Gavage
DuPon-15302
Ondo 530A Pg "aTnhleycilcbaaltsitoanndcaordvsenwashegeterrenadlstbyanFdegrs(sfioen1n0aFilgs 1s)i.nDghaepfeoarkeastesoaluitiofnrsowerehe Compared othecalibrationcurvesto cvalatth concataionsof he 24921
"(TeCsVt.s=ubssttaanncdearudrdifvoirsmtitiyolnetahneve3h1ic0l0e owfatsheevmaeiasstuerdedbycocanlecauliaatiinogntshocfoehfefdiucpielnitcoaftvsearmipsloens v(asiifaioonrotfoefsmtihnxoricunalg01c0%overniisftcchaetseitoasnn)datrodrcrcracaohdtnosoiitnHgoalesvkneell.lLAacbooreaftfoircyifeonorft accepaobf helestesubdstasncerroebghoutthtesoatinon. "0Thdeemteen etshlocofncheenctratcionomf hce ectsuabstatnsceamifpolretoshe(r3ne=sp2ev)ctfoirvrcedaocfshdocslienvgealesv.etwaos wnid `VSeubriliittyownassavmplleasadbhyewbiasneglitnheefo eacnomspaartiongfttheh orfesoproindyionfgsmianbiiglConecsenst.raion
THe
PAlmamsomnaiCuomncePnetrrfaltuioroonocintaPnocast-eW:eaAngiengERfafscFtotlohlnoewing Oral Gavage
DuPont 15302
Da155f0 Aogee
ANALYTICALRESULTS
A. Chromatography ap2p4r5o2i1aleltyd3f.r5moimntuhteeHsPL2DCcLolinjection8s)rfeosrctvheedneegastkiwvietihona.rReetpnrteosnenttaetivoefLC-MS/MS cContrho. TrhiswoaasrmdsehtoeawrmnintneFdbigyorescgo2mp(arrai)so.noTfethseubdisltuaennctewwaes nwotidhaencdiwditnhotu 0tme/mal. standardagain he Omg.contro containing internalstandard. RetoFfiguere 2r0 ).
B.RecoverySamples Dcoencaetdrasiolnoaft2r4e9s21sfoofrrtehceov2.erymsga/mmpLl.esevaerlewsuamsm9a7i.z3%eodifnoTnaibnleal..TThhee mecaassureedd c{obneceanntalryattiicoanlmoeftHho2d4p9e2r1ffoorrtmheedsa7t.i5smfagc/tomriLlyeovevlrwthse n10i2e.1c%oonfcennormianiaoln.aBnagsedoornttheissdyt,
. Uniformity of Mising/Concentration andStabilitySamples
Anailfyotrimciatlyorfesiussinrgo/mCodnicnengratsiionovnesfccouliloecnteado-nhAuagurso,o20t0e4mspnedrsantaelsyazebdior ae
Shown iAppeTaable IdadiSuriamary<Tb,|.
"stTahbeilfiotlylaowninygasblFeosr uhmimssaarmipelsintghedryeosfutshfeuo Hs-2i4f52o1prrempiartatyiocn.oncveenfitcatrioan aonnd
Sel ema Wael SoVmeres T(V5): Nema
sm5iasCpoienmion 0 so
= wm
5iS asmeaemn whiess 33 mwma
+b VS D esru oi efoetop ed.Rcel pmootfehrems Gpheisrw.seabmse.cpuionof eitws ith
Frnoazhe-oesi io nd Con 1 ed idwin. Sr
{1"2eT4shsterstuhbuasnts1a0nf)coerwnadF-ts2b49tl2h1enasatmghpeleveesdhpiCrcoelnpecawernhetdersnatnhideolncdsol(l=Be1oc6tu.erd9os%noAof onuromstieam9lp,2e.r0a0d8eiqnudftioeclalytmeedivtxeheltdst(heCV Tetsubstancewa notdeedinth 0mpmsamples.
E. Conchusion sRubesstoalnscerwoatsmeiaknealypsriosopfertyh(eCH-V2l4e9s2s1thdaonsi1n0g)s,oluthioeotnsaforctieesvtsdy(i+n2di0c%atoeftaotmhee)esta.nd
abe derthecondofotemstosdy.Testsbstsoewasnokfound he 0gl.samples.
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PAlmamsomaniCuonmcePnetrrfaltoiroonocitnaPnoosstte-:WesAngiengERfaftecstFoonlltohewing Oral Gavage
DuPon-15302
Deis Acres
STuubpee.Becoery of n24g921Aid2de2dt1oDoin Vei= ce a Tommi Mossad Noman oTRecrony ge245 n16S ion ed A. 1 Bip arn npefdmoeitnparrepnionoe f svery ge.Thecg covey plese
Tite0.SUase ofMapCosaSVeermeinnsfSul of 2472iDigTSohnion.
Shiopmeor em
Nem
em
UCno to a
son
-
"n
a2ss
Ma:
m2 1103
hfod aissx)
a"
723s
1iv5a
slasx
Mei: 8o5i7i2016 ro
Son Tr i,Bp
2d53s5
i2i006 i niiits
T iderroeS mne ace a rhrtmi (pSroec,eeta odie,hic ol.
SHEET mop.
-_--
PlAamsmmoaniCuonmcePnetcrfaltuioornoocintPanoosatt-eW:eAangiengERfaftscFtotlohlnoewing Oral Gavage
DuPont-15302
Dn 1530 A ge
`ReprAesasFiivgeiurcen)CatlibarattioniCuvrvees:
ae mt es
`Equationforthe fted line:
Y= 035+39126.5478 *7X
0 ReSquared = 0997699
3
|
i
|
.
om | ob ok | om obs os Concentration, ppm
Figure 1: CafvoiibtrchaaNltieibprooantpciuuornrevslwschetoeiworisnaognfldim2neaa2xi70inc3e)h2(ePor6eFvp5lOeic3AactnoeanlpcyeetaniickraralStetioannadtriaardgs)sqduoaferesd) 9005100051 ppm.
eer
PAlmamsomaniCuonmcePnetrrfaltuiooronoicniaPnoosatt-eW:eaAngiengEfRfacstoFnoltlhoewing Orsl Gavage
Dap 1530AN get Figure
Representative LOMSMSChromatography Chromatograms
DuPont 15302
Fig 2a: Rselpy.rsNcetaaiieeLnOPSFOSAhrroeatsoignatmoefNiaosppraorisonwicilry(3.5mmpais.ea) ed ae.
fte ywith emsada.Negative 55 mins.
Pe FOArtnins ie apprvaiienlyoe
7
PAmlmaosniCuonmcePnetrrfaltoiroonosfnanPcossteW: eAigenEfRfaetts Foonltlhoewing Or Gavage
D15ue ae Fi2gcountiroused)
ReprLOe MS/sMSCehron matt ograa phyt Chri omatv ogre ams
Duron 15302
= Rtepietnaikne.LONMeSgchievSesPtRogO2 A cn pnfyn o 3of5Ec AcTiswitht Fie0 R(Ee30mpapiwniLlChNSmSevosrsmotgedmoOf3127p0e5n16N5ePeFOAAFsFaOAyriScinoinirr)e
pperi3y3 ines. - =
PAlmamsomaniCuonmcePnetrrfalutoiroonocitnaPnooast-eW:eaAngineg
Effect Rats
on the Following
Oral
Gavage
Don15302 AYPage 10
Fi2g(cuontrinueed) `RepresentativeLOMSMSChromatographyChromatograms
DuPont-15302
Figure2:cRoenpcrenernuaiiovnoefL0O03Spcnhroofrsi2o4g9r2a1moffo1r.c5mgagieiodoiPnFOsAowluittihoniceaidntio peo3i.5s mite. ThemeserecothneFecseertntnasltaiinio.n65mogf.
FL igure3:ReprsetciervaeLMSs chio= miogram ofigh coverydosing sluion (1.45m/tie)died i1mmsinpalrcoorni3ac.5iemieoonfult0te0ys3.rTphpaemmotefaisi3or4e7dn2c1ofnocneesgraatiiovneofonhePrFeOpArweistehnrtestEieconvsy scionis 71 men 395
AmPmlaosnmaiCuomncePnetrrfaltuioroonocintPanoosatt-eW:eaAngiengERfaftscFtotlohlnoewing Oral Gavage
DuPont-15302
sep Ae ap, 20e 4
nt
Tiel
`SSumsyopfDoseing nioAssnes
DagComdn ibyoo2n (ent
N25
us
rot
"
aannny
awsss
n
awnmy
woos
AvAeverraaggeeMPeerwcaereNdoCmiosnac
aasns
aaens)
SCaonndteierntDoefvVorna'in
rwy
sIoi
oRyn Tepe
20
in
an
asm
T? DN ded ucn edn Rcepth w penee eepclta ofmpeedwdd eies..S6uwB a Fevisl.i w ed Teeenn meeme scs.nie ofen spit,itddein, lio
a0
PAlmamsomaniCuonmcePnetcrfalutoiroonocitnaPnoosstte-:WeaAngiengEfRfatcstoFnoltlhoewing Oral Gavage
DuPont15302
Appendix B PFOA Plasma Concentration Data - Males
EE
PAlmamsomsniCuonmcePnertfrlsuioornoocintaPnooast-eW:esAngiengEfRfaectt oFonltlhoewing Oral Gavage
Duont-15302
Time Post
PFOA Plasma
Group (wAeegkes)
(Dhoousrse)
NAunmimbaelr
(Dmgo/skeg)
Conceniation g/mL
Mean
SD
1
3
2
I
10
20 as a0
1
3
2
2 10
4430
1
3
2
3
10
39.80
1
3
2
4
10
4119
I
3
2
6
10
3685
Tr
3
2
8 30
3895 12065 1278
m
3
2
10
30
123.62
ji]
3
2
2 30
104.66
ut
3
2
15
30
113.53
i]
3
v
T
2
16
30
2
51
10
122.48 3537
92 44 _
v
4
2
5 10
a3
v v
4 4
2 2
5 58
10 10
3585 4554
v
4
vir
2 2
61 52
io 30
39.70
~
179 740 1810
vii 4
2
55
30
8641
vii 4
2 36
30
131.53
vii 4
2
57 30
27.73
vit 4
2
& 30
12403
IX 5
Ix
5
2
9
10
2 100 10
2036 %32 68 37.00
IX
5
2 103
10
217s
x
5
2 105
10
2.09
X Xi
5 3
2 107 io 2 9% 30
2039 5963
666 155_
Xi x
5 5
2
97
30
2 9% 30
951 58.10
xi
5
2 101 30
71.36
x1 5
2 102 30
89.69
2s
PAlmamsomnaiCuonmcePnetrrfaltuiooronoicntaPnoosat-eW:eaAngiengERfaftscFtotlohlnoewing Oral Gavage
DuPont-15302
Group T 1 1 1 1 ur ut nt ui
om v v v v v vii vit vii vii vi xX xX x x x Xi
Xi Xi Xi xi
Time Post
PFOA Plasma,
Age Dose Animal Dose Concentration
(weeks) (hours) Number (mpg) nl) Mem
3
2%
9
10
3523 un
3
2 13
10
21.00
3 3
2 2%
1 18
10 10
36.13 23
3
22
10
3649
3
2 7 30
6530 76
3
2 19
30
7680
3
u 2 30
8005
3
u zn 30
98.90
3
2 2s 30
4973
[]
22
10
4004 CII
4 4
2 x 65
10 10
268 4732
4
23
10
4885
4 2 7 10
35.80
T 4
2% ot 30 u 6 30
1207 10081 106.94
4
u 8 30
108.70
4 4
u 7 30 2 7 30
96.60 79.64
5 2% 108
10
37.05 W060
5 5
2 109 EA 110
10 10
39.60 052
5
Bi
ns 10
3905
5
2 iis 10
4680
5 5
2% 106 30 u ni 30
98.30 386 98.70
5
u n 30
103.53
5 5
2 13 2 ua
30 3
153.89 11488
SD 78 183
1308 36 2336
_
TT
PlAamsmmoaniCounmcePnetrrfaltuioornoocintaPnooast-eW:eaAngineg ERfafetcst oFnoltlhoewing Oral Gavage
DuPont15302
Appendix C PFOA Plasma Concentration Data - Females
--
a ----
APlmamsomnaiCuonmcePnertfrlsoiroonocitnaPnooastte-:WeAngiengEfRfaetcst Foonltlhoewing Oral Gavage
DuPont-15302
"Time Post
PFOA Plasma
Group
(wAegekes)
(Dhoousrse)
Animal Number
Dose Concentration (mg/kg) g/ml) Men
SD
[
3
2 26
10
20.64 387 ST
on
3
2
27
10
37.87
[i
3
2 31
10
4590
n
3
2 33
10
3863
n
3
2 38
10
373
Ww
3
2
5
30
96.95 886 1050
wv
3
2 30
30
89.64
v
3
2 2 30
7441
v3
2 3s 30
90.13
v3
2 36
30
7.18
vi vi
4 4
2 2
7
10 10
27.69 288 11s 3105
vi vi
4 4
2 2
7 n
10 10
1927 991
vi 4
3
8 10
21.48
iif E] vit 4
2 76 30
2
8 30
103.54 8067 1410 67.98
vi vin
4 4
2
30
2 ES 30
70.38 79.15
vin
4
2
84 30
$228
x x
5 5
2 122 10 2 24 10
44.00 BB 74 2.66
x x
5 5
2 126 10 2 127 10
33.88 30.08
x
5
2 128 10
3451
XII 5 xi 5
: ny 30 2 ng 30
7817 5690 29.66 9861
pt 5 xi 5
2 120 30 2 21 30
3555 3486
xu 5
2 12s 30
3730
_
Ts
PAlmamsomaniCuonmcePnetrrfaltuiooronocintaPnooastie-:WeaAngiengEfRfeactt oFonltlhoewing Oral Gavage
DuPone15302
Age TimDoesPsost Animal Dose PCFonOcAenPilaatsimona
Grow (weeks) (hous) Number (mye)
ml) Men SD
[
3
2%
I
10
[5] 1355 38
n
3
x
"
10
1370
n
3
x
6
10
1376
i
3
x 49
10
1938
i
3
2 50
10
1208
v
3
2 37 30
3625 Sia el
wv
3
pl 39 30
51.05
wv
3
pl 2 30
229
wv
3
2 pe 30
7143
Iv 3
2 I 30
3613
vi fl
u 79
10
1663 98 700
vi 4
2 81
10
25.20
vi 4
28
10
24.04
vi
4
2 91
10
767
vi
4
2 94
10
239
vil E 2% 86 30
37.70 B/OI 990
vin
4
2
7
30
3931
vin 4
2
88
30
21.62
vit 4
i
9
30
17.32
vit 4 2 9 30
2009
X
5
2% 130
10
249
[ET
x
5
u 31
10
092
%
5
u 132
10
036
x
5
2 133
10
1.00
%
5
2 137
10
202
XI
5
2 129 30
061
342 195
xu
5
2 134 30
553
xu 5 2% 135 30
433
xu 5 2 136 30
433
xu 3 2 138 30
230
Ts
PAlmamsomnaiCuonmcePnetrrfalutoiroonocitnaPnooast-e:WeaAngiengEfRfacstFoonlltohewingOrsl Gavage
DuPont 15302
Appendix D `Supplemental PFOA Plasma Concentrations
-_--
APlmamsomaniCuonmcePnetfrlautoiroonocitnaPnoosatt-eW:eaAngiengEfRfaetcst oFonltlhoewing Oral Gavage
DuPont15302
PFOA Plasma Concentration
Animal
(ng/ml)
Fasted Male
8428
Fasted Male2
562
Fasted Female
s820
Fasted Femal2e
wis
NonFasted Male 3
2981
Non-Fasted Male 4
3020
Non-Fasted Female 3
3194
Non-Fasted Femal4e
2014
Note: Awlelreanciomlallesctweedr2e43hwoeuerskpsosotfadgoesinagn.d receiasvineglde 10
Mean 6495 616 3000 2654 gi dose.
sn 234 110 028 764 Plasma samples
---
PlAamsmmoaniCounmcePnterraftlioornoocitnaPnoosstte-:WeaAngiengEfRfatcstoFnoltlhoewing Oral Gavage
DuPont-15302
Appendix E Urine PFOA Concentration Data - Males
-Ts
APlmamsomnaiCuonmcePnetfrlautoiroonoicntPanoost:eW:eaAngiengEfRlaetcstFoonltlhoewing Oral Gavage
DuPont15302
Animal
CPonFcOeAntrUartiinoen
Group Age (weeks) Number Dose (mgh) (ug/ml) Mean
1
3
9
10
926
957
1
3
13
10
1621
1
3
1
10
341
1
3
18
10
197
1
3
x
10
697
ur
3
7 30
2.69 5176
ut
3
19 30
71.90
ut
3
2
30
80.97
m
3
2
30
NS
m
3
2
30
3048
v
]
@
10
on
5
v
4
10
362
v
4
65
10
545
v
4
66
10
585
v
4
0
10
701
vii
]
6
30
1632
570
vii
4
6
30
255
vii
4
8
30
1099
vii
4
7
30
5817
vii
4
7
30
3547
IX
5
108
10
278
0
x
5
109
10
404
IX
5
no
10
235
Ix
5
ns
10
285
IX
5
16
10
810
XI
5
106 30
Xi
5
ni 30
20
i565
1375
Xi
5
2 30
2561
x
5
us 30
1746
x1
5
ua 30
10.16
NS = 00 urine sumple was produced by the animal.
SD. 436 2586 . 245 884 236 on
THs
PAlmamsmoaniCounmcePnetrrfaltuiooronocintaPnooastie-:WesAngiengEfRfeacttoFnoltlhoewing Oral Gavage
DuPon-15302
Appendix F Urine PFOA Concentration Data - Females
--_--
PAlmamsomnaiCuomncePnetcrfaltuioroonocintaPnoosatte-:WeAangiengERfaftscFtotlohlnoewing Oral Gavage
DuPoni-15302
PFOA Urine
Animal
Concentration
Group Age (weeks) Number Dose (mgkg) (agin) Mean sD
1
3
[0
10
1759
217 895
1
5
"
10
1936
n
3
10
1004
it
3
9
10
un
It
3
50
10
3411 >
v
3
3
30
116.09 9489 2636
wv
3 39
30
8367
wv
3
a2
30
95.85
wv
3
5
30
56.68
wv
5
4
30
122.17
vi
7
10
521
526 1527
vi
4
81
10
7.16
vi
4
8
10
679
vi
4
91
10
4737
vi
4
%
10
1379
vir
T
86
30
12555 Toe 2897
vin
4
8
30
6035
vit
4
8
30
9084
iit
4
90
30
13116
vit
4
92
30
112.68
x
5
130
10
4104
77 266
x
5 131
10
2385
x
5
132
10
89.76
x
5
133
10
5289
x
5
137
io
4131
XI
5
129 30
99.53
m6 5136
xn
5
134 30
lL
xn
5 135 30
55.41
xu
5 136
30
175.44
xu
5 138
30
17425
Tas