Document E0MDed0JVyxd5pgnzNwEnv3L
COVANCE
"UNAUDITED DRAFT
August 20, 1998
Sponsor:
3M St. Paul, Minnesota
FINAL REPORT
Study Title:
4-Week Capsule Toxicity Study with
`Potassium Salt (PFOS; T-6295)
Perfluorooctane Sulfonic
in Cynomolgus Monkeys
Acid
Author:
Peter J. Thomford, PhD
Study Completion Date:
"To be determined
Performing Laboratory:
Covance Laboratories Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704-2595
Laboratory Project Identification:
Covance 6329-222
`Sponsor Project Identification: 3M Study No. T-6295.6
Page 1 of 212
02408
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CovamnMcTeT6e2320s0.5s226s2
COMPLIANCE STATEMENT
4-WecPoktCaaspssiuulmeSTaoltxi(cPiEtOySS;tuTd-y62w9i5t)h PinerCfylnuoormooolcgtuanseMSounlfkoenyisc Acid
`AGllooadspLeacbtosroafttohriysPsratcutdiyceweSrteanidnaradcsc,or4d0anCcFeRwi7t9h2.the Environmental Protection Agency
SPtetuedryJD.iTrheocmtofrord, PhD Covance Laboratories Inc. A-- ndrew-- M_ Seae cat, PhD Study Monitor 3M
Date -- Date A------
2
02409
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CovawnScrMeT6a362s299-s252e62
QUALITY ASSURANCE STATEMENT
`This report, with the exception of Appendices 6, 7, 8, and 9, has been reviewed by the
Quality Assurance Unit of Covance Laboratories Inc., in accordance with the
4E0nvCirFoRnm7e9n2.talThPerdoatetcatiionn AApgpeenncdyic(eEsP6A,)7,Go8,odanLdab9owrearteorgyePnrearcattiecdeSbtyanadnadradrse,
responsibilityofthe were conducted and
Sponsor and the findingsreported
Sponsor's designees. to the study director
The and
the
followinginspections study director
mmaannaaggeemmeenntt.acWcroirtdtienngsttoatsutsarnedpaorrdtsopoefriantsipnegcptrioocnesduarneds.findings are issued to Covance
Tnspection
--
Date Reporied to
Dates
From
0421/98
To
04/21/98
Protocol
Phase
Review
Study Director and Study Director Manag nt
04721798
05/20/98 05/20/98 Protocol Amendment Review
05120198
"
05/20/98
05/21/98
05/20/98
05/21/98
Protocol Amendment Review
Clinical Laboratory Inspection
05/20/98
05/21/98
06/01/98 07/08/98
06/01/98 Postlife 07/13/98 Data Review
06/01/98 07/13/98
08/14/98 08/14/98 Protocol Amendment Review
08/14/98
08/10/98 08/18/98 Report Review
08/18/98
Representative
Quality Assurance Unit
Date
3
002410
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CovanMcTeT6a2320m0-5s2262
STUDY IDENTIFICATION
4-WeePoktCaaspssiuulmeSTalotxi(cPiFtOySS;tuTd-y62w9i5t)h PinerCfylnuoormooolcgtuanseMSounlfkoenyisc Acid
Test Material Sponsor Study Monitor Altemate Study Monitor Study Location Study Director
Study Timetable `Study Initiation Date IInn--LLiiffee T(eErxmpienriamteinotnaDla)tSetart Date Experimental Termination Date
PeSraflltuo(rPoFoOcSt;anTe-S6u2l9f5o)nic Acid Potassium
3M BTuoixlidcionlgog2y20S-e2rEv-i0c2e,s 3M Center St. Paul, Minnesota 55144-1000
A3nMdrew M. Seacat, PhD 612.575.3161
3MaMrvin T. Case, DVM, PhD 612.733.5180
3C3o0v1anKciensLambaonraBtoourlieesvaIrndc. Madison, Wisconsin 53704-2595
PeterJ. Thomford, PhD
Covance P.O. Box
Laboratories 7545
Inc.
Madison, Wisconsin 608.241.7207
53707-7545
April 17, 1998 April 23, 1998 May 22,1998 To be determined
4
002411
KEY PERSONNEL
CovanMceT6632299.52262
Study Director Study Toxicologist Study Coordinator Supervisor, Large Animal Toxicology Supervisor, Dose Formulation Supervisor, Laboratory Animal Medicine
Clinical Pathologist
Supervisor, Clinical Pathology Anatomical Pathologist Supervisor, Anatomical Pathology
PeteJr.Thomford, PhD Dale Aldridge, BS
Patricia K. McKee Pesik, BS, LAT Meechelle Bordeaux, LAT Dixie Bushee, BS, LATG DDiopnlnoamJa.teC,lAemCoLnsA,MDVM, MS RDiopbleormtaLt.e,HaAllC,VDPVM(C,liPnihcaDl Pathology) Ronald Markevitch, BS,MT (ASCP) `Thomas E. Palmer, PhD Kimberly W. Durland, BS, HT
5
002412
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0 TCounR Tce66392S 952262
CONTENTS
Page
Reserve (Archive) Samples... rms 12
TAENGFCAON. corres,
"HOUSING ANG MBIRLENANCE ....ccrsssrss
JUSUFICALON covers essen rss
SS ------------
13
sss: 13
Group Designations and DOSE PLEPATBON wc.
DOSE
LEVIS wero. sss:
14 14
Methodof Administration................. sss: 16
BClOinYicaWlEOIbGsHeSrvc atio onss.....t .....s .............. runsanna
snanaes sens nennsnssssssasssasssnnnnen 16
Rectal Body TEMPERUIES ......sssss
17 17
Blood Hormone Determination...................... rss
17
Serum PFOS Level
CHClPAthOIORY
Deters mination....s ....... r sss
17
18
NAEddCiTtOiPoSnYalc Blood COr lleCtion .t ..............v.vrvoreenn rms
18
19
Palmitoyl CoAOxidaseDeterminations ........................... EE --
Cell Proliferation EVAIation........................ uremsnsssssssssssasssasssasusasasasssasssasssasasae 19
Liver PFOS Determination........................ rissa
19
HTiISSSULE OPIPEASENIOVAIOON.ZYocct rersssst sneessss mses
oeerererssmennirrsee 19
20
SHALSHCAlADBIYSES......cccvsmmssssso
20
DOSE ARAIYSES ....coocoeeer rrr
r-------- sss 33
Clinical ObSErvations .......................... rs
sss sessssssis 2.
Body WEIGhLS ...c.oceeveevererssssssssssnsenen rms 33
02413
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ComvaTnMceeT66m32290s-522s62
CONTENTS (Continued)
RESULTS (Continued)
Page
ReFcOtOaClOBMOSYUTMPEOMN Pr ETUIs ES ss rs
22 22
SBe1r0u0mHPORMOMSONLGevAelNGDEItYeSMUEASU.O.N..c..c.ocvo.vsosvomsssmesssm CHinical PAROIOBY wos
23 23
CLeVlElrPPrORGOCSTaDteitonetEmiVatAioInU. An ON.e ..e .ovr coes ssemsss
23 ee 23
24
ADRUOCE PAOIORY corse
24
CONCLUSIONS tse 24
PATHOLOGYREPORT ccc, 21
COM ONTM HEDAE TA..N . TS
30
COGDeEnSer,aAl CBodBesRanEdVABIDIAEATVINILDIOUONNNISTSS,v .....s ....coe nsmmssns
31 32
AC bbrevd fioartCiloinS nsiacnadl UPniAtstfoOrClIinOicaBl YHE.MA.OIcOg.Ysore oo
33 35
`ACbobdreesvfioartiAoNnOs MaIndCAUnlitPsUfONrOClIinOicRalYC..H..E.MvoIeSreTomYs.me.e.res
37 30
TABLE
1 2
SummaryofClinical SummofaClirnicyal
OObbsseerrvv-a-3aAt0Mt.iM/iioWnEuonEtKesnslYsP.os.t.d..o.s.e.............o.o.o.r.r.n4410
3 4
Summary ofClinical SummofaClirnicyal
OObbsesrevartvi--oan9t60s0 iMMiionnuuntteesss
PPOOSStdSoEse.........o......44.32
5 7
SS ummu aryoOm ffBCloidnm iycWaelia OghBtDr SatEa y rVUna(SCKtHgEi)A.UoI.E.nd s
oo.44 45
Summary of Rectal Body Temperature Data (-C)...ovoeooos
8 9
Summaryof Clinical SummofaClirnicyal
HHemeatmoalotgyoDDaltaao-t~gDDaaayyy2-7 9.o .. o
47 49 53
10 11
SummofaClirnicyal Summaryof Clinical
ChemDiatse t-DArYy-7 Chemistry Data - Day
overs 2...
57 63
1132SSumumamrymooffaCClliirnniiccyaall CChemhisEtrymDDaattiaa--SDDaa(yy 71r.....y. 14 SummaryofClinical Chemistry Data - Day29...
69 75
15 16
ISnucimdmeanrcyeooffCMlaiCnriocaslcoCphiecmOiBsStErIyVDAaUOtNaS.- .D.a.y........u
30.........oom or
81 87
89
17 IncidenceofMiCroScopic OBSEIVAUONS wove
90
7
002414
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CovanwcIeMrT6e.362m209s.526s2
CONTENTS (Continued)
APPRPOELNOCDOIIXDEL VIRONS o ...ns rssers smssme smeosos sssen, so P9a03g3e
PPrPRIOOOOIMOCOCOCO!!O AAMc EMNIEMEt ENt NNOO..s 2 Lo m ss ommm es s es
03 113
MPartOeOrCiOa]l ASafMetEy DEat!a SNHO.EL3....
remeron
115 117 120
API InPdiEvniNduaDdl2ICAXRiCc D v]FORBiA SEEDIAdVA] Oc NaS clcvon orossa smrmss, oson,121122786
Individual Rectal Body Temperature Data (-C).......ovmrevooenoomonso APPENDIX 3 coosvnransssssssessssmsmssssssssssssmssmsmsossosnn,
133 131
Individual Body Weight Data (Kg) ..vecrversmsmrersoensnso APPE4cNoDvIX senesmsssissssssssensssssssssssssssmsossonnnn
138 140
IIndinvidudalCCliliinniivccaalliCHHeEmMdaItSoTluYogyaDaltdD.at.a....v.....ve.oe.m.smorrmeowsomesoeensosoooen1s5. 1341
APIPnEdNSA.DomisaIslvvXsPnOirhnOsIdsOGsYsDusanstamRs.o.lm.s.s.ss.svsesosssmsessmsrnrsesssomesoosesnesosnen1sn8n, 6185
APSPuEmNmaDrIy6Xandc Indivo idual c HOMMOo NG ARo AIYSEn S Datas ... Figure 1'- MeanEStrDaatda-iol Males...
snore
198 199
FiFgiugurree 32 -- MMeeaannTEsrtriadiiooldDaotat-hFDyEaMrtAaloE-snM.Bi.I.En.S.e..............o.mo.e.ro.o.so.o.oonnosonoorn.
203 204 205
Fig4u-Mreaen Triiodothyronine Data -FEmales........o...ooooonoror. 206
APPEND7cIXcs
201
APSPeErNuDmIaXnd8Lc iverPFOs SLevel Ds eterminations oe... 210209 APCePllEPrNODU9FITXAOcNcEovVcAsUssAsOsNs.ss.scs.ecmmcarmrssrsesmssssmsssimmosssossmssensosssmosmsmnnn221121
8
002415
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CovanmIcMeTT6-3a6220s-9252s62
ABSTRACT
m`aTxheipmuurpmosteoloefrattheids sdtousdeyawnadsltoowperrodvoisdee ldeavtealsfotrodbeeteursmeidniinngaacnhreosntiicmattoexidcity study and (foPFaOssSe;ssT-t6h2e9e5f)f,ecotnofcrtihteictaelsetnmzatyemreiall,evPeelsr,flhuoorrmoooncetsa,neaSnudloftohneirc sAecliedctPeodtbaisoscihuemmiScalatl parameters.
GMraoluepan1,dtfhermeealaenicmyanlosm/oselxguisn Gmroonukpey2s, awnedreonaessainginmeadl/tosetxhrieneGgrroouupps3)(.twAonainmiamlaslsi/nsex in w(Gerioguhpts/d2ayan(dmg3/krge/cdeaiyv)e,d rgeelsapteicnticvaeplsyu,lteisucroantteadinwiintgh0l.a0c2toasned(21.:09P9FaOnSd/k1:g9,owfebyo,dy troetsaplecmtaitveerliya)l. dAosneimlaelvselifnorGrcoacuhpgIrroeucpeiwvaesd 2g0el.a0timng/ckagp/sdualye.s containing lactose only. The
wFeoroedowbasserpvreodvitdweidceondacielyor(at.wmi.ceanddailpy..m.W)faotremrowrtaasliptryovainddedmaodrilbiubnidtiutmy.. TAhtelaenasitmaonlcse.
cdaoinlsyu,mapntiimoanlswwaesraesesexsasmeidnqeudalfiotrataibvneolyr.malIintaidedsitainodn,siagnnismaolfstowxeircietyo,bsaenrdvfeodod
baephparvoixoirm.atBeoldyy30w,e6i0g,htanddat9a0wmeirneuctoelslepcotsetddtowsiecfeowreseikgnlsyobfepgiononrihnegal2thweoerkasbnboefromrael
tinhietrieaatfitoenr.ofRetcrteaatlmbenotd,yotnemDpaeyra-t1u,roens twheerefirrsetcdoarydeodfttwriecaetmwenete,klayndbetgwiincneiwnege2klwyeeks
ebsetfroiorle,itnhiytrioatiidonstoifmutlreaattimnegnth.ormBolnoeo,d tsraimipoldeosthfyorronhionrem(oTn3)e,aannadlytsheysro[xesitnra(dTi4o)l], ewsetrreone,
tchoellseactmepdlebsefwoerreeinsietnitattioonAonfitLryetaitcmseInntc.a,nfdorpraenadloysseeso.n DBaloyo2d9.samSpelreusmfworassehraurvmested, and
DPaFyOsS2,c3onc(eanptprraotxiiomnataneallyy2s4eshwoeurrse acfotlelrecttheedfbrestfoarnedisnietcioatnidondoosfet,rreeastpmeecnttivaenlyd),pr7e,do1s4,e on
and 29. samples
Serum was harvested, and samples were collected for hematology and
were sent to the Sponsor for analyses. Blood clinical chemistry tests before initiation of
treatment and on predose on Days
Day 2, 7,
29. and
In addition, blood for 14. Additional blood
clinical chemistry tests was was collected at the time of
collected
`eOxnsaDnaguyin30a,titohne,aannidmaslasmpwleerse waenersethsehtiipzpeedd, twoeitghheeSdp,oenxssoarngfuoirnpaotsesdi,blaenfdutnuerceroapnsaileyds.is.At
necropsy, macroscopic observations were recorded, and selected tissues were collected
'
cc2416
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CovanMcTTe e6632w290s.52262
paenldmiptroeyslerCveodA. oIxniaddadsietiaocnt,ivaitsyaamnpalleysoefs.livReerprweassenctoaltlievcteesdafmrpolmeseaocfhliavneir,matelstfeosr, and.
fpoarncprroelaisfewreateiocnoclellelctneudclferaormanetaicghenaneivmaalulaatinodn.senAt stoamPpaltehoolfogliyveArsswoacsiactoelsleIcntteedmfatrioomnal
ecxaacmhiannaitmiaolnsanwderseendtotnoetohne Stpheonasdroernaflosr,PeFyOe,Sfceomnocreanltrbaotnieonmaanrarloyws,esl.unMgi,clriovserc,okpiidcney,
pancreas, spleen, testes, and thymus from each animal.
eAflflecatnsimoanlcslisnuircvalivoebdsetrovtahteiosncsh,ebduoldeydwseaicgrhiftisc,e.fo`oTdhecroenwseurmpetnioont,esrtemcattaelrbiaoldyrelated temperatures, clinical pathology, or macroscopic or microscopic findings.
sTthiemruelawteirneg hnoorampopnaer,enotretshtyrmoaxtienr.iaTlh-ererleatweedreeffneoctaspopnareesnttrotenset,measttreiroill,-ttheylraotiedd effects on
estradiol or riodothyronine at 0.02 study, itis not possibletodetermine
mg/kg/day. Given the conclusively whether
low number of there was a test
animals
in
this
fmeatmearliealgirevleant2e.d0emffge/cktgo/ndaeystwreadrieolnuomretrriiciaodloltyhylroowneirnet.haEnstthroasdeioolfltehveelcsonftorrotlheanmiamlaelsanodn
f`Doamyal2e9gainvdenth2e.i0r rmegs/pkegc/tdiaveywperersetunduymevarliuceas.llyTrlioiwoedrotthhaynrotnhionseeloefvetlhsefcoornttrhoelmaanliemaalnsdon
vDaalyue2s.9;Throiwoedvoetrh,yrtohneiinperelsevteuldsyfvoarlauneismwaelrsegailvseonconosri0d.e0r2abmlgy/lkogw/edraythwaenrtehenucmoenrtiroclally
haingihmearlsongiDvaeny22.90cmogm/pkagr/edadywwitehrethneuirmeprriecstauldlyy lloewveelrscwohmiplaerterdiiwoidtohthtyhreoinripnreelsetvuedlys for
levels
+Inwceyenkos,motlhgerueswmeornekneoysaptpraeraetnedt wteistthm0a,er0i.0a2l,-roerla2t.e0dmegffePcFtOs So/nkcgl/idnaicyaloroablsleyrfvoartiaotnlse,ast pbaotdhyolwoegiyghftisn,difnogosd. cTohnesuomnpetimoanl,eraecntdalobneodfyemtaemlpeetrraetautreeds,wiotrhc2l.i0nimcagl oPrFOaSna/tkoem/idcaayl aanpapleyasreesdatnodhcaevle plroowleifrelreavteilons oefveaslturaatdiioonls ownillDabye p2r9.oviDdaetda bfyrotmhelSivpeornasnodrsaenrduPmaPthFoOlSogy `Associates International, respectively.
10
002417
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CowvanMTcTe.e60322s09-25s262
PURPOSE
`The purpose of this maximum tolerated
study dose
was to provide and lower dose
data for determining levels to be used in a
an estimated chronic toxicity
study
and
(toPFaOssSe;ssT-t6h2e9e5f)f,ecotnofcrtihteictaelsetnmzatyemreialle,vPelesr,flhuoorrmoooncetsa,neaSnudloftohneirc sAecliedctPeodtbaisoscihuemmiScalatl
`parameters.
REGULATORY COMPLIANCE AAlglenascpyecGtosoodf LthaibsosrtautdoyrywePrraecdtiocneeSitnanadcacrodrsd,a4n0ceCwFiRth7t9h2e.Environmental Protection
TEST MATERIAL AND VEHICLE `Test Material LTohteNtoes.t2m1a7t,eriisala,wPheirtfelutooroofofc-twahnieteSuplofwodneirc.AcItidwaPsotraescseiiuvemdSaatltC(ovPaOnSc;e To-n6295), September 4, 1997. The test material was stored at room temperature. tIhnaftordemfaitnieonthoentessytntmhaetseirsiamletishoodns,filsetawbiitlihtyt,hepuSrpiotyn,socro.mposition, or other characteristics
Vehicle
(`Tehxepivreahtiicolnedwataes Fleabcrtuosaery(S1p3e,c1t9r9u9m),. NIet wwaBsrurnescewiivcekd, aNt eCwovJaenrsceey)o,nLMoatrNcoh.
NNO192 30, 1998.
The
vehicle was stored at room temperature.
Information on synthesis the vehicle is on file with
methods, purity, composition, the manufacturer.
or
other
characteristics
that
define.
un
002418
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MrCoevanIcmMeT6.s362209.s5262
ARerseesrevreve(sAarmchpilvee)(1S0agmpealcehs)ofeach lotofthe test material, vehicle, and each test
tmeamtpeerriaatlu/raec.toTsehetsrietusraamtipolneswawsiltlabkeentbreafnosrfeeirnrietditaotitohne oSfpotnrseoatrmaefntteracnodmsptloerteidoantorfotohme
in-life phase.
RDeismpaoisniitniogntest material will be retained at Covance for use in possible future studics.
TEST SYSTEM
YTeosutnAgnaidmualltot adult cynomolgus monkeys were obiained from Covance Research
Products initiation
Inc. (Alice, Texas), of treatment.
on
March
10,
1998.
The
animals
weighed
2.010
2.5
kg
at
EIdaecnhtiafniicmaatliownas assigned a permanent number upon arrival and identified with collar tag before initiation of treatment. All data for an animal are recorded under this number,
SAcecvleinmmaatlieosnand seven females were received on March 10, 1998, transferred from the
RCoovoamnc2e01i5n-fhooru3se8 sdtaoycskcboelfoorneyionnitiMaatirocnhof16t,re1at9m9e8n,t.andInacgecnleirmaalt,eadniinmaAlnsiminalthis
shipment appeared healthy. abnormalities indicative of
During acclimation, the animals were examined for health problems. In addition, three tuberculosis tests,
a
physical examination, and a fecal flotation for parasites were done on cach animal,
a`Tbhneorpmhaylsiitciaels.exRaemsiunlattsioofn tdhoenteuabfetrecrultohseisantiemstaslswewreerenergeacteiivveedforreavlelaalneidmnaolsm;atjhoerresults nareegarteicvoerdfoerdpianratshietedaotav.a. TOhneeremsaulltes aonfdthoenefefcaelmaflleotawteiroen etelsitmsiinnadtiecdatferdotmhsetaundiymals were consideration based on clinical pathology findings or abnormal fecal observations during
12
002419
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CuovranMceTe-6m63229s9.52s62
1a0ccglriomuaptsio(nt.woThaenriemawlesr/esesxixinmGalreosupan1d, tshirxefeeamnailmeaslsin/stehxe irnanGdroomuipza2t,iaonndfoorneasasniigmnamlesnetx.
in Group colony.
3).
Animals
not selected
for
the
study
were
transferred
to
the
Covance
stock
AHnoiumsailngRaonodmM2a0i1n5tweansanucseed for this study. Environmental controls for the animal room
were set 12-hour
to maintain 18 to 29EC, light/12-hour dark cycle.
a relative humidity of 30 to 70%, Variations from these conditions
and are
a. documented
in
the
data and are considered to have had no effect on the outcome of the study.
The animals were housed individualilny stainless steel cages.
Certified
numbers
primate diet
are recorded
(in#3t7he26dCa,ta.HaTrlhaendTieetkliasdr)ouwtaisneplryoavniadleydzeodncbey
or twice daily. The lot
the manufacturer for
anuntdrictoinotnaalmicnoamnptonaneanltysseasnadreenovnirfoinlemewnittahlCcoovnatnacmei-nMaantdsi.soRne.suFlrtusitosfosrpeacdidfiiteidonnaultrient
supplements were provided, but did not require analysis.
m`Wiactreorowrgaasnpirsomvsiadneddaednlviibriotnumme.nta`Slacmopnlteasmoinfatnhtes.watTehraerreesaunltaslyazreedofnorfilsepewciitfhied Covance-Madison.
There were no interfered with
known contaminants this study.
in
the
diet
or
water
at
levels
that
would
have
Justification
nPoFnOhSumisaankpnriomwantehsep(astuicchpaesrtohxeicsyonmoemporlolgiufsermaotonrk(ePyP))rienstphoenrdats.imWilhaerlnyetxophousmeadntso PP,
(.e., Tow species.
to
no
hepatic
response)
and
therefore
are
an
appropriate
human
surrogate
13
002420
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CovaunMcTTe 66e322m99-s5226s2
PROCEDURES
PTrhoitsosctouldAymweansdcmoenndtucNtoesd.in1 atchcroorudghan3c.e TwihtehptrhoetoPcroolt,opcrooltodcaotledaAmpernidlm1e7n,ts19,9a8n,dand protocol deviations are in Appendix 1
SGerloecutpioDneosfigannaitmiaolnssfoarntdhDostsuedyLewvaeslsbased on clinical observations, clinical pathology, `acnodmpouttheerridzaetdabalsocakppirnogprpiraotcee.duArneidmeaslisgnweedretoaascshiigenveed btoodtyrewaetimgenhtt gbraoluapnsceuwsiitnhagrespect to treatment group.
Group
Dose Level (mg/kg/day)
Total Material Dose Level (mg/kg/day)
Number of Animals Males Females
T (control)
0
2 (low dose)
0.02
200
2
2
200"
3
3
3
(high The
dose) control
group
20 (Group
1)
received
200 an equivalent
1 amount
of
Tactose
1 in
b Tgehleatlionwc-adpossuelegsroasupthreectoetiavlemdattheeritaelstamdamtienriisatletrreidtutroatGerdowuipth3.lactose
(1:99, ww).
The 9,
high-dose ww).
group
received
the
test
material
riturated
with
lactose
GDeolsaetiPnrceappasrualteiso(nTorpac, Inc. Fairfield, New Jersey), Size No. 2, Lot No, 122630 r(eixtpuirraatteidownidtahtelaJcutnosee2a6n,d20l0a2ct)o,seweornelyu(sceodnftorroldogsreouapd)miwneirsetrdaitsipoen.nseTdheintteostcampastuelreisaltwice weekly and stored at room temperature. bVeahsiecdloe.n tLhaectveohsieclweasasussuepdplasietdhefovreGhircoluepin1.the dose preparations. Dose levels were
1
002421
_--
CovaSnMcTei.663e2295:m25262
For Group 1 dose preparations, the specified amount of lactose was weighed and
transferred into the gelatin capsule. The top and bottom halves of each capsule were
joined, and the
`number, group
capsule was placed into the appropriate container
number, animal number, and dose level.
labeled
withstudy
`Test Material. For Group 2 and 3 dose preparations, the test material was dissolved in
acetone and triturated with lactose (1:999 and 1:9, wiw, respectively) once before
initiation of treatment. Acetone (Spectrum, Gardena, California), Lot No. LH0253
(expiration date June 2000) was used to dilute the test material
with lactose was necessary to facilitate capsule preparation.
with
lactose.
Trituration
For Group 3 dose preparations, the specified amount of test material was weighed, placed
into a labeled mixing container,
container. The preparation was
and the appropriate volume of acetone
stirred manually until the test material
was added to the
was dissolved.
`The specified amount of lactose was weighed, placed into the `mixing container, and thoroughly mixed using a spatula. This preparation was exposed to air until theacetone
was evaporated and stirred periodically. Samples of the finished preparation for dose
analyses were
transferred to
moisture.
atackoenntadiinreerctalnydfsrtoomretdheatcornotoaimnetre.mpTerhaetutrrietuarnadtepdrtoetsetcmtaetderfiraolmwlaisght
and
For and
Group placed
2 preparations,therequired amount of the Group 3 preparation was measured into a labeledmixing container. The specified amount of lactose was
weighed, placed into the mixing container, and thoroughly mixed using a spatula for
approximately 5 minutes. Samples of the finished preparation for dose analyses were
taken directly from the container. The triturated test material was transferred to a
container and storedatroom temperature and protected from light and moisture.
`The specified amount of triturated test material was weighed and transferred into the
gelatin capsule.
was placed into
The top and bottom halves of each capsule
the appropriate container labeled with study
were joined, and the capsule
number, `group number,
animal number, and dose level.
i
02422
_--
MrCoevanImcMeT6-s362299.s5262
D`SoasmeplAensa(l1ysgecsach) were collected from the top, middle, and bottom of the test
mtaetmepreiraalt/ulraec.tosTeheprseapmarpalteisonwserfoersthhieplpoewd-uanndedrhaimgbh-ideonstecgornodiutpisoansndtosttohreeSdpaotnrsooormfor
analysis.
pHroemsotguedynestiatbyilsitaympanlaelsyscios.lleOctneed sfertoomftshaemmpildedsl(e1ogfetahcehp)rewpearreattiaoknesnwferroemutsheedlfoowr-tahned tPhiegShp-odnossotresftormaantaeliyasilsl.ctose preparations at the end ofthe inlfe phase and shipped to
Results of the Sponsor are in
homogeneity, Appendix 7.
stability,
and
dose
confirmation
analyses
provided
by
the
MGeeltathiondcaopfsAudlmeisnwiesrterautsieodnfor test material preparation administration to compare with odfateaxpforsoumrep.revious toxicology studies using the oral route, which is the most likely route
(T7hedadyossfevepcrke)pafroarti2o9nsdawyesre(saeedmPirnoitsotceorledDeovriaaltlyioinnsgfeolrateixncecpatpisounlse)s.onTcheeddaoisliyng tubes
ownerteheflmuosshtedrewcietnhtl5ymreLcoorfdreedvberosdeyowsemiogshitss,wawtietrh.thIendeixvciedputailondoosfesbowdeyreweciaglchutlcaotleldecbtaisoend
days when the previous same time each day.
body
weight
was
used.
Animals
were
dosed
at
approximately
the
"CTlhieniacnailmaOlbssewrevraetoibosnesrvedtwicedaily (a.m. and p.m) for mortality and moribundity. aEbancohramnailmfailndwiansgsobwseerrevreedcdoaridleyd,.anAdnifmoaoldscownesruemopbtsieornvewdasapapsrsoexsismeadtqeulayli3t0a,ti6v0e,lya;nd. r90ecmoinrudteaesds ptohsetydwoesreefoorbsseirgvnesod.fpAonoirmhaelasltwheroer oabbsneorrvmeadlobnecheavwieoer.klyE;ffaecbtnsorwmearlefindings or an indication of normal was recorded.
16
002423
-_--
CovamnIcMTeT6e-362s29-s9252s62
Body Weights
iInnidtiivaitdiuonalofbotdreyawtmeeingtht(MdaotnadwaeyraenrdecTohrudresddatyw)i,coenweteheklfyirsbtedgaiynnoifngtr2eawtemeenkts, baenfdortewice
weekly thereafter. calculations.
An
additional
body
weight
was
recorded
on
Day
-1
forthe
Day
1
dose
RReeccttaallbBooddyyteTmepmepreartuarteusrweesre taken at approximately 11:00 twice weekly beginning f2owreeexkcesptbeifoonrse).iniAtniiatmiaolnsofwetrreeantomtenatne[sMtohnetdiazyedanfodrTthhuerpsrdoacyed(usreee.Protocol Deviations
Blood Hormone Approximately 5
Determination mL of whole blood
were
collected
from
a
femoral
vein
of
each
animal
bbeeftowreeenin0it7i:a0ti0onanodf 0t9r:e0a0t.menAtni(mDaalys-7w)earendfapsrteeddosoveeornniDghatyb2e9f.orBelcooolldecwtaisoncso.llAelcltesdamples
cwleotr.e cSoalmlpeclteesdwweirtehocuetntarnitfiucgoeadguwliatnhti,nma| ihnotuarinaefdtratcrololoecmtitoenm,pearnadtusreer,uamnsdaamlplloewsewdetroe
Dhaaryve2s9teodn.lyS)earnudmswtaorseddiivniadefdreienzteortswetotaopmparionxtiamiante-l6y0etqou-al80alCiquunottisl (pbalcokoeddcoolnlrecyteidcoen
easntdrosnhei,pepsetrdio(lo,AtnhiyrLoyitdicsstiImnucl.a,tfionrgahnoarlymsoense.,Tthreiisodaomtphlyersonwienree(Ta3n)a,lyazneddtfhoyrroexstirnad(iTo4l),.
:
Results of these analyses were provided by Ani Lytics Inc., and are in Appendix 6.
SApeprruomxiPmaFtOeSlyL5ev(eDlaDye-t7eornmliyn)aotrio2nmL of whole blood were collected from a femoral
(vaepipnroofxiemaacthealnyi2m4alhobuerfsoraeftienrittihaetifoinrsotfatnrdesatemceonntd(dDoasye,-r7)esapnecdtipvreeldyo)s,e7,on14D,aaynsd22,9.3
aAnntiimcaoalgsulwaenrte, mfaasitnetdaoivneerdniatghrtoboemfotreempceorlalteuctrieo,nsa.ndAlalllsoawmepdletoscwleotr,e Scoalmlpelcetesdwweirtehout
cfreenetzreirfusgtedtowimtahiinnta1ihno-u6r0atfote-r8c0ollCecutnitioln,paacnkdesdeornudmrwyaiscehaarnvdesstheidpapnedd tsototrheedSipnoansor
for analyses. The samples were analyzed for PFOS. `Sponsor are in Appendix 8.
Results of analyses provided by the
17
002424
--_--
CovanMMcTTe -6a36e229w9-25s262s
BCllionoidcaslamPpaltehsolwoegrye collected from each animal on Days -7 and 29 for hematology and
`celaicnhicaanlicmhaelmipsrterdyosteestosn. DIanyasdd2i,t7i,ona,nbdlo14o.d fAonricmlailniscawlecrheemfaisstterdyotveesrtnsiwgahts;cwoaltleecrtwedasfrom
provaidldibietudm. evaluated:
Blood
was collected
from
the
femoral
vein.
The
following
were
Hematology
red blood cell hemoglobin
(erythrocyte)
count
hematocrit
`mean corpuscular volume
`mmeeaann ccoorrppuussccuullaarrhheemmoogglloobbiinn concentration
platelet count
`white blood cell (leukocyte) count
difsfeegrmenetnitaledblnoeoutdrcoeplhlilcocuonutnt Iymphocyte count monocyte count eosinophil count basophil count rbeltoiocdulcoeclyltemocropuhntology
glucose urea nitrogen creatinine total protein albumin globulin total bilirubin cholesterol triglycerides aalsapnaritnaetaemaimniontortarnasnfsefrearsaese alkaline phosphatase
Clinical Chemistry `sograbmitmoal gdleuhytdarmoygetnraansseferase creatine kinase calcium sinoodriguamnic phosphorus potassium chloride serum bile acids amylase lipase pancreatic-specific amylase
`AWdhdoilteiobnlaolodBl(oapopdroCxoilmleactteiloyn20 mL) was collected from the vena cava of each animal at
WthaestpiomteaosfsieuxmsaEnDgTuiAn.atiSoanm(pSleeesPwreorteoctorlanDsefveirarteidoinnstofocronetxaceipnteirosn(sa)p.prTohxeimaantteilcyoa5gumlLant
ach) and shipped to
stored in afreezer set the Sponsor possible
to maintain -60 future analysis.
to
-80
C
until
packed
ondry
ice
and
18
002425
--_--
CmovaTnMceeT6m2320s0-52s262
Necropsy
xOynlaDzianye,30w,eiagnhiemda,lsbltehadtfwoer rreeqfuaisrteedd toevstesr,niegxhstanwgeurineaatneeds,thaentdizneedcrwoiptshiekde.taAmniinmeaalnsdwere
necropsied in random order.
oTrhifeicneesc;rtohpesycriannicalludceadviatym;atchreosbrcaoipnicanedxasmpiinnaalticoorndo;ftthheeneaxsatlemcaalvistuyrafancdepoafratnhaesbaoldy; all
sinuses; viscera.
cervical
tissues
and
organs;
and
the
thoracic,
abdominal,
and
pelvic
cavities
and
Palmitoyl CoA A sample of the
Oxidase Determinations right lateral lobe ofliverwas
collected
from
each
animal
at
scheduled
sacrifice. `maintain
The sample -60 10 -80C
was until
flash-frozen in liquid nitrogen andstoredina analyzed for palmitoyl CoA oxidase activity.
freezer
set
to
RCeeplrlePsreonltiafteirvaetisoanmpElveasluoafttihoenliver, testes, and pancreas were collected and preserved in ezvianlcufaotrimoanliwne.reAfetmebrefdidxaetdioinn,psaarmapffliensafnodr psrhoilpipfeerdattoioPnatcehlollnougcyleAasrsoacnitaitgeesn I(nPtCerNnAat)ional IfnotrePrnCatNiAonaelvaalrueatiinoAn.ppReensduilxts9.of the evaluation provided by Pathology Associates
ALivsearmpPlFeOofSlDiveetrewramsincaotlileocnted from each animal at sacrifice, weighed, flash-frozen in ilciequainddnisthriopgpeen,d waintdhsptolraesdmainsaamfpreleezsertsoetthteoSmpaoinnstoarinfo-r6P0FtOo S-8a0naClyusnitsi.l pRaescukletdsoonf tdhreyse analyses provided by the Sponsorare in Appendix 8. Tissue Preservation pTrheesefrovleldowiinng10ti%ssnueeust(rawlh-ebnufperreesedntf)oormrarleipnr,eusnelnetsastiovtehesrawmipsleesspweceirfeiecdo,llfeocrtpeodssainbdle future microscopic examination:
19
02426
--_--
adrenal (2) aorta brain cecum cervix colon duodenum epididymis (2) esophagus eyes [preserved in Davidson=s
fixative (2)] femur with bone marrow (articular surface of the distal end) hgeaalrltbladder
ijleejuumnum
kidney (2) liver lung `lmyammpmhanroydgel(amnedsenteric)
CovanMmceTr6232s09.5s2262
muscle (thigh)
ovary (2)
pancreas
pituitary
prostate
rectum
salivary gland sciatic nerve
[mandibular
(2)]
seminal vesicle (2)
skin
spliunamlbacro)rd (cervical, thoracic, and
spleen
sternum stomach
with
bone
marrow
ttehsytimsu(s2)
thyroid trachea
(2)
with
parathyroid
urinary bladder
uterus
vagina
THhisetaodpraetnhalosl,ogeyye, femoral bone marrow, lung, liver, kidney, pancreas, spleen, testes, and etxhaymmiunsewdermieceromsbceodpdiceadlliyn rpaoramffeian,chseacntiimoaneld(,sseteaPirnoetdowciotlhDheevmiaattiooxnyslifnoraenxdcecpotsiionn,sa)n.d
Bone marrow smears from the sternum of prepared, stained with Wright's stain, and
each animalatthe scheduled sacrifice retained for possible examination.
were
SOtnaltyisdtaitcaalcAolnlaelcytsedeson orafterthe first day of treatment were analyzed statistically.
Statistical analyses of animals.
were
not
done
on
data
collected
for
Group
3
due
to
the
small
number
2
002427
_--
CovaMnScTMeT6e362w299.s252s62
wOenieg-htwsa,yraenctaallysbiosdoyftveamrpiearnacteur[eA,NpaOlVmiAto(yWliCneorA, 1o9x7i1d4a)s]ewacatsivuitsieeds,taonadnacloynzteininuiotuiasl body
clinical pathology values.
hLeetveernoeg'esneteistty(oLfevveanreia,nc1e96a0t)p w<as0.d0o5,netrtaontsefsotrfmoartivoanrsiawnecreehuomsoegdetnoeisttayb.iliIznetthheecvaasreiaonfce. sAigNniOfiVcAantw,aDsundnoenteto'nt-ttheesth(oDmuongneente,ou1s96o4r)twraasnsufsoerdmefdordcatoan.troIlf tvheersAuNs OtrVeaAtedwagrsoup. comparisons.
wOenieg-htwsa,ywaintahlyisniitsiaolfbcoodvyarwieaingchet[sAasNtChOeVcoAvar(iWaitnee.r,Al1t9h71obu)g]hwLaesveunes'esd tteosatnfaolryvzaerbiaondcye ahdojmuosgtemneenittryewmaosveddoneext(rsaeneeaobuosvhee)t,enroogetnreaintsyf.ormIfattihoenAs NweCrOeVusAedwabsecsaiugsnieficcoavnatr,ilaenacset Squares means ttest was used for control versus treated group comparisons.
For each sex, Group 2 was compared with Group 1 evaluated at the 5.0% two-tailed probability level.
(Control).
Group comparisons
were
RECORD RETENTION
tAhlilsastwuddyatwai,lldboecuamrencthaitivoiennd, trheecosrtdosr,agperoftaocciolilt,ieasnodfsCpoevcainmecnes-Mgaedniesraotnedfoarsaapreersiuoltd ooff f1inyaelard.isOponseityieoanorfafttheretmhaetesruibamlsi.ssiAloln roafwthdeatfainsatlorreepdorotn, mthaegnSeptoincsomredwiilaladnedtearnmiorniegitnhael copy of the final report will be retained by Covance-Madison.
(W2istahpipnro1pryieaatrea)ftferrosmutbhmeisSspioonnsoorf'tshdeefsiingalnereespo(rt3,MalElTo.f t&he,afAonriemLeynttiicsonIendc.maatnedrials
Pathology Associates Intemational) (Andrew Seacat, PhD, 3M).
will
be
sent
to
the
Sponsor
21
02428
_-- RESULTS
Dose Analyses Results of analyses provided by the Sponsorare in Appendix 7. Results of the dose analyses will be provided by the Sponsor.
CovanMMceTr62e329m9.5s226s2
Clinical Clinical
Observations observations are
summarized
in
Tables
1
through
S;
individual
data
are
in
Appendix 2.
All animals survived to the scheduled sacrifice. clinical observations.
There were no testmaterial
related
Body Weights Body weight data are summarized in Table 6; individual data are in Appendix 3 There were no apparent test material-related effects on body weights.
Food Consumption CFloiondiccalonOsbusmeprvtaitoinondast-aAa.reM.s/uWememKalry)i;zeiinnddiTvaibdulael1da(tSauamrmeairnycloufded in the individual clinical observations in Appendix 2. `There were no test material-related effects on food consumption.
Rectal Body Temperatures
Rectal body Appendix 2.
temperature
data
are
summarized
in
Tables
7;
individual
data
are
in
There were no test material-related effects on rectal body temperatures.
2
002429
_--
CovanMIcMTeTa6-3e6292s-92s52s62
`BSluomomdaHroyramnodninediAvniadluyalsehsormone analysesdataare in Appendix 6. Estradiol and
triiodothyronine analyses data are illustrated in Figures 1 through 4 (Appendix 6).
sTthiemruelawteirnge hnoorampopnaer,enotrttehsytrmoaxitne.riaTlh-reerleatweedreefnfeotaspopanreesnttrotneest, measttreiroila,lt-rheylraotiedd effects on
estradiolor study, itis
triiodothyronine at 0.02 not possible to determine
mg/kg/day. Given the conclusively whether
low number of there was a test
animals
in
this
fmaetmearliealg-irveelnat2e.d0emffge/cktgo/ndaeystwraedrieolnuomretrriiciaoldoltyhylroowneinret.haEnstthroasdeioolfltehveelcsonftorrotlheanmiamlaelsanodn
fDeamyal2e9gainvdenth2e.i0r rmegs/pkegc/tidvaeypwreersetunduymevraliuceasl.lyTrlioiwoedrotthhaynrotnhionseeloefvetlhsefcoornttrhoelmaanliemaalnsdon
`vDaaluye2s9.; Throiwoedvoetrh,ytrhoeniirnperelsevteuldsy fvoarlauneismwaelrsegialvseon c0onosri0d.e0r2abmlygl/odwearythwaenrtehenucmoenrtircolally
ahniigmhearlsongiDvaeny22.90cmogm/pkagr/edadywwitehrethneuimreprriecsatluldyylloewveelrscwohmiplaerterdiiwoidtohthtyhroinripnreelsetvuedlys for
levels.
Serum Results
PofFsOeSruLmevPeFlODSetleevremlindeatteiromnination
provided
by
the
Sponsor
are
in
Appendis
5.
Results oftheserum PFOS level determination will be provided by the Sponsor.
HCleimnaitcoalloPgaythaonldocglyinical chemistry data are summarized in Tables through 15;
individual data.
data
are
in
Appendix
4.
The
Pathology
Report
contains
adiscussion
of
the
Administration ofPFOS had no effects on clinical pathology test results.
RCeeslullPtrsoolficfeelrlatpiroonliEfveraaltuiaontieovnaluation provided by Pathology Associates Intemational are in Appendix 9.
2
002430
--_--
CovanuIcMerT6-e362w29-9s252s62
Results of the International
cell
proliferation
evaluation
will
be
provided
by
Pathology
Associates
Liver PFOS Determination
Results of liver PFOS level determination provided by the Sponsor are in Appendix 8
Results of the liver PFOS level determination will be provided by the Sponsor.
Anatomical Pathology aInncdid17e.nceIsndoifvimdaucarlodsactoapiarceainndAmpipcernodsicxop5i.c TobhseerPvaatthioolnosgyarReespuomrmtacroinztaeidnsinaTdaibslceusss1i6on of the data.
There were no test tissues examined.
material-related
macroscopic
or
microscopic
changes
in
the
organs
and
CONCLUSIONS 4Inwceyenkos,motlhgerueswmeornekneoysaptpraeraetendt wteistthm0a,t0e.r0i2a,l-orlrat2e.0dmegffPecFtOsSo/nkgc/lidnaiycaloroablsleyrfvoartiaotnlse,ast pbaotdhyolwoegiyghftisn,difnogosd. cTohnesuomnpetimoanl,eraecntdalobneodfyemtaemlpeetrraetautreedsw,iotrhc2l.i0nimcagl oPrFOanSa/tkogm/idcaaly aanpapleyasreesdatnodhcaevlel plroowleirfelreavteilosnoefveaslturaatdiioonlsownilDlabye p2r9.ovDidaetda bfyrotmhelSivpeornasnodrsaenrduPmaPthFoOlSogy Associates Intemational, respectively.
2%
02421
-_--
Covance 6329-222 aM3TMTe6w29s5s6
SIGNATURES
--_---- Patricia K. McKee Pesik, BS, LAT Study Coordinator Covance Laboratories Inc. Peter J. Thomford, PhD Study Director Covance Laboratories Inc.
F-- re ---- Date Date
=
0 2432
_--
CovaMnIcTMe 6aT322w99-s5226s2
REFERENCES
Dunnett, C. W., "New Tables for Multiple Comparisons with a Control," `Biometrics, 20:482-491 (1964).
SLteavteisntei,csH,.(,e"dRso.bLusOtlkTienstestfaolr,ECqhu.al2i5t,ypopf. V2a7r8i-a2n9c2e,s,S"tCaonnftorridbUuntiivoenrssitotyPrPorbeasbs:iliSttyanafnodrd, California (1960).
i`nWiEnxepre,rBi.meJ.n,t"aDleDseisgingna,ndSeAncaolnydsiEsd.o,fCShi.ng3l,ep-pF.ac1t4o9r-E2x6p0,erMicmGenrtasw,-"HiSltalt:istNicealwPYroirnkci,ples New York (1971)
SWeinceorn,dBE.d,J.,C"hA.na1l0y,spips.of75C2o-v81a2r,iaMnccGer,awSt-aHtiisltli:calNPeriwncYioprlke,s iNneEwxpYeorrikme(n1t9a7l1bD)e.sign,
2%
02433
-_--
CovanM MceTe6T32290.5s 2262
PATHOLOGY REPORT
SUMMARY m`aTxheipmuarposteoloefrattheids sdtousdeyawnadsltoowperrodvoisdee dleavtealsfotrodbeeteursmeidniinngaacnhreosntiicmattoxeidcity study and (0PFaOssSe;ssT-t6h2e9e5f)f,ecotnofcrtihtiecatlesetnmzatyemreialle,vePlesr,flhuoorrmoooncetsa,neaSnudloftohneirc sAecliedctPeodtbaisoscihuemmiScaaltl `parameters.
Atedsmtimnaitsetrriaatli-orenlaotfePdFmOacSrohsacdonpoicefofrecmtiscroonsccloipniiccalchpaantgheosloignyttheestorrgesaunlstsaannddticsasuusesed no. examined.
METHODS m`Tahtreereiaglrooruaplslyofvimaaclaepsaunldefaetmaaldeosceynloevmeololgfu0s (mcoonntkreoylsgrwoeurpe;ardemcieniivsetdetrheedvtehheictelset, glarcotuopseh),ad0.t0w2,ooarni2m.a0lms/es/ekxg, othfebloodwy-dweoisgehgtr/oduapy (hmagd/tkhgr/edeaayn)ifmoarls2/9sedxa,ysa.ndThtheechoingthr-odlose `group had one animallsex.
tBrleoaotdmewnats(cDoalyle-c7t)eadnfdorohneDmaayto2l9o.gyInanadddciltiinoinc,alblcohoedmifsotrrcylitneistcsalbcehfoermeisitnrityitaetsitosn wofas cmoalclreocstceodpoincDoabsyesrv2a,t7i,oansndwe1r4.e Trehceoradneidm,aalnsdwteirsesuseascrwiefriecepdraensdernveecdrionpsfiixeadtoivneDaas y 30; hseppeactiifciepdablmyitthoeylprCotooAcool.xidSaasmeplacetsivoiftyliavnedr wfeorredectoelrlmeicntaetdiaonndoffrPoFzeOnSfocrondteetnetrm(ibnyattiheon of fSopronmsaolri)n.foSraemvpallueastoifolnivoefrp,rotleisfteesr,atainodn cpealnlcnruecalsewareraentcioglelnec(tPedCNaAnd; pbryesPeartvheodloignyzinc bAossnoecimaatrersowIn,telrunnagt,iolniavle)r,. kMiidcneryo,scpoapnicrceeaxs,amsipnlaeteino,ntseswteesr,eadnodntehoynmuasdrfernaolms,alelyaen,ifmaelmso,ral
Results of clinical statistically.
pathology
tests
for
the
control
and
low-dose
groups
were
compared
2
02434
-_-- RESULTS AND DISCUSSION
Mortality All animals survived to the scheduled sacrifice.
CowvaTnIcMeeT6-3w6229s-0252s62
Clinical Pathology
Individual values and
values for hematology and clinical chemistry ests the resultsof statistical comparisons are in Tables
are in Appendix 8 through 15.
4.
Mean
aDbanoyrm7a.litCliiensi.cal pathology test results indicated no obvious group or individual health
cDhaeymsis2t,r7y,taesntdre1s4u.ltsAdamitnhiessteractoliloenctoifonthientteesrtvamlast.erial had no effects on clinical
Statistically significant differences low-dose groups were inconsistent
for clinical chemistry results between the over time and between the sexes. Several
control of the
and.
edxihffiebrietnecdeas swterroengsidmoislearretloadtiifofnesrheinpc(eis.ep.,retsheenhtiagthD-daoyse-7a,nainmdalnsodniedonfottheexdhiifbfietrneontcaebsle
differences for the same parameters).
Day 29. Administrationofthe test chemistry test results at Day 29,
material
had
no
effects
on
hematology
or
clinical
cSotautnits,tiacnaldlyhespiagtniicfipcaalnmtidtiofyflerCeoncAesofxoirdaesde wbleoroedicneclolnsciosutnetn,tnbeeuttrwoepehnilthceousretx,esmoannodcyte aelxkhailbiinteedphnoospdhoasteasreelawteiroensshiipm.ilaSrtattoisdtiifcfaelrlyensciegsniofbisceanrtveddifafterDeanyce-s7.for triglycerides and
Anatomical Pathology oAfnattheommiaccarlospcaotphiolcoagnydfmiindcirnogsscofporiceaocbhsearnviamtailoanrsearieniAnpTpaebnldeisx156. anIndci17d.ence summaries
2
002435
_--
CovanMcTTe 66a32200m.522s62
t`iWseseueks 5e.xaTmhienreed,wearnedntohemraecwreorseconpoicmaajnodrfdeiwffmeircenrcoesscobpeitcwefienndicnognstrionl tahnedotrrgeaantsedand
animals in incidental
the and
incidence unrelated
of to
any finding. All the test material,
microscopic
observations
were
considered
_-- Robert L. Hall, DVM, PhD.
--_-- Date
Diplomate, ACVP
Clinical Pathology
--e `Thomase E. Palmee r, PhD Pathologist
Date
29
002436
-_--
CovaMnMceT6T 632995s 2262
(COMMENTS ON THE DATA
Vdaatraioinusthmios dsetludsy.of Bcealccauulsateodrsi,fecroemnptutmeordse,lsanrdocuonmdpouftfeorr ptrroungcraatmesnwuemrbeerusseddiftfoeraenntallyy,ze
svlailguhetslyinfrsoommtehotasbeleisn o(ct.hge.r, mtaebalness,,fsrtoamndianrddidveivdiuaatliloyncsa,locruliantdeidvidadtuaa,l ovralfureosm) mstaatyisdtiifcfaler
analysis data. differences.
Neither the
integrity
nor
the
interpretation
of the
data
was
affected
by
these
Some tabular data were compiled using Excel' Version 7.0 software.
`The summary condition was
tables for observed
clinical without
observations regard to the
indicate specific
the number of animals for which nature, severity, reversibility,
a
numberofincidences/animal, othe length of time the condition persisted.
Only observations tables.
other
than
normal
are
indicated
on
the
summary
clinical
observations
`Theday of initiation of treatment is "Day 1."
30
02437
-_-- 2
CODES, ABBREVIATIONS, AND UNITS General Codes and Abbreviations Codes for Clinical Pathology
Abbreviations and Units for Clinical Hematology Abbreviations and Units for Clinical Chemistry
Codes for Anatomical Pathology
CowvanITcMeTe6-362s29-92s5262
Note: STohmeef,olbluotwniontgnleisctessosfacrioldyesal,l,aobfbrtehviisaitnifoonrs,maatnidonunmiatsyabree nueseeddebdyfCoorvtahnisce. report.
31
02438
_-- WK N Mean; MEAN CAM SD; S.D.; STAND DEV; STANDARD DEV; sd NA P UNSCHED DISPATCH
MIN [=
General Codes and Abbreviations
CovanIcMeTT6-e362s92.9s25262
Week. NAruimtbhmeertiocfmmeeaans.urements in a group. CSotvaanrdiaartde-deavdijautsitoend. mean.
Gfrroomup 2thmeemaenanisofsitghnieficcoannttrloyldgirffoeurpent
(Group 1) atp #0.05.
No value; Present
not
applicable;
not
present.
Unscheduled.
Obsmeordvualteioonfstthreandsaftearrceodllferctoimonthseyisnt-leimfe:to
ntehecrnoepcsryo.psOybsmeordvualteiofnosr arreefedruepnlciecadtuersionfg
Mitnhuetela.st in-life observations.
Observation.
Animal T
Death
Codes:
Terminal sacrifice.
32
002439
--_--
Ns QS/QNS NR FS sc SH H SL L st 1 NF u DT/DOT DB I] TE RE EE SE PC PD PI PL PA co HB PLASMO NO AGG FR UTD NO COAG
Codes for Clinical Pathology
CovanmMcTeT-6e3622w99-s2526s2
GENERAL CODES
No sample
`Quanity not sufficient
No repeat (sample Fibrin strands
volume
not
sufficient
for
repeat
analysis)
Sample clotted
Slightly hemolyzed
Hemolyzed
Slightly lipemic
Lipemic
Slightly icteric
Ioteric
Animal not fasted
AUnnismchaeldduileeddo/mnotreistbund bleed
Died during bleeding
`TTeecchhnniiccailanerjruodrg(miennsttrtuomernetpeoart tteescthnician error that results in
supnialclceed,pteanbtlrey odaftian,vea.lgi.d, duantaac)ceptable instrument output, sample
sRpeeclolridnginegrreorr,roirn(correrceocrtdeddatien)correct data, .g., wrong number,
Entry error (incorrect Sampling error
keyboard
entry)
Platelets clumped
Platelets decreased
Platelets increased
Platelets large
CPloaltoerleitnstearpfpeeraesr waidtehqutaetste
Heinz bodies observed
Plasmodium
No aggregation
Fractious
Unable to determine
No coagulation
3
02440
-_--
CovaM nIcMeT6T -36229.9s 25262
Codes for Clinical Pathology (Continued)
RESULTS NOT INCLUDED IN STATISTICAL ANALYSES
``SHaemmpolleyszfedrocmlianinciamlaclsheamtiusntsrcyhoerducolaegduilnatteirovnalssamples APcrtoitvhartoemdbpianrttiiamletshr(oPmTb)ogprleaastteirntthiamne5s0(sPeTcTo)ndgrseater than 110 seconds Bleed times (BLETIME) greater than 30 minutes
(CODES FOR BLOOD CELL MORPHOLOGY Tpohiekifloolclytoowsiinsg (sPcaOlIeK)w,aspoulsyecdhrtoommaesaisau(rPeOtLhYe)d,eghryepeocohfraonmiasosciyato(sHiYsP(OA)N,ISaOn)d,toxic neutrophils (TOXNEUT):
Scale
Degree
B Normal for the species
1 Slight
2 Moderate
3 Marked
--_-- 4 Not ape plicae ble
Presence Not present Rare Few Moderate MMaanyy
34
002441
--_--
CovianMceT66a322w9-5s226s2
Abbreviations and Units for Clinical Hematology
Test
Red blood cell count
Hemoglobin
Hematocrit
Mean corpuscular volume
MMeeaann ccoorrppuussccuullaarrhheemmoogglloobbiinn concentration
Platelet count
Mean platelet volume
Reticulocyte count
HAebisonlzubtoedryetciocuunltocyte count
PErryotthhrroocmybtiensetdiimmeentation rate
Activated Thrombin
partial time
thromboplastin
time
FiAbcrtiinvaotgeedncoagulation time
PFliabtreilne/tfiabgrgirneoggaetniodnegradation products
Collagen
Adenosine diphosphate
Alpha 2-antiplasmin
Bleeding time
Methemoglobin
Plasma hemoglobin
EMsyteilomiadt/eedrmyytehlrooiidd/reartyitohroid ratio
White blood cell count
DifNfuecrlenetaitaeldbrleododblcoeloldccoelulntcount
Corrected white blood cell count
`Segmented neutrophil count
Band neutrophil count
Lymphocyte count
Monocyte count
`Eosinophil count
AnBiassoocphyitlosciosunt
Polychromasia
Abbreviation (Units)
RBC
HGB
(E6/UL
(G/DL)
or
X10%0L)
HCT (%)
MCV (FL)
MMCCHHC(P(G%))
MPLPTV((EF3L/)UL or X10/L)
RETIC (%)
HREETIINCZ ((%E)3/UL or X10*/L)
ESR (MM/HR)
PT (SEC)
PIT (SEC)
TT (SEC)
FABCRT((MSGEC/)DL)
FDP (UG/ML)
PAGG/COL (%) PAGG/ADP (%) ANTIPLAS (%) BMLEETHTIGMBE((%S)EC) PLA HGB (MG/DL) ME RATIO EST M/E RATIO
WBC (E3/UL or X10/L)
NRBC (/100 WBC)
COR WBC (E3/UL or X10/4L) N-SEG (E3/ULorX10%/L) and % N-BAND(E3/ULor X10"L) and %
LYMPH (E3/UL or X10%/L) and %
MONO (E3/UL or X10%L) and %
EOSIN (E3/UL or X10%L) and %
ABNAISSOO((E13/2U3Lo)r X10%L) and % POLY (-12,3)
002442
35
_--
CovanuIcMerT6-e36229s-925262
Abbreviations and Units for Clinical Hematology (Continued)
Test Poikilocytosis Hypochromasia Howell-Jolly bodies Basophilic stippling `Toxic neutrophils AquAteyopuiscawlhiltyempbhloocoydtecesll count (right eye) `Aqueous white blood cell count (left eye)
PAbObIrKev(i1a,t2i3on) (Units) HYPO (123) HBAISBTOIDPY(1(21.,32)34) TOXNEUT (-,1,2,3,4) ATYPLYM (1.234) REYE (WBC/UL) LEYE (WBC/UL)
002443
36
---
wCoTvaneIcMeTs6-362s29-02s5262
Abbreviations and Units for Clinical Chemistry
Test
Glucose
Urea nitrogen
Urea
Creatinine
Total protein
Albumin
Globulin
Albumin/globulin ratio
Total bilirubin
Direct bilirubin
Indirect bilirubin
Cholesterol
Triglyceride
TUorteaal nliitpridosgen/creatinine ratio
Phospholipids
LHoiwgh--ddeennssiittyylliippoopprrootteeiinncchhoolleesstteerrooll
Uric acid
AAlsapanritnaetaemaimniontortarnasnfsefrearsaese
Alkaline phosphatase
S`oGrabimtmoal gdleuhtyadmryolgetnraansseferase
CLarcetaattienedkeihnyadsreogenase
Amylase
Lipase
Palmitoyl CoA oxidase
Calcium
Tonized calcium
Inorganic Sodium
phosphorus
Potassium
Chloride
Magnesium
Zinc
Strontium
Tron
Abbreviation (Units) UGLNU(M(GM/GD/LD)L) UREA (MG/DL) CTRPERAOT(G(/MDGL/)DL) ALB (G/DL) GA/LGORBA(TGI/ODL) TBILI (MG/DL) D BILI (MG/DL) IBILI (MG/DL) CHOL (MG/DL) TRIG (MG/DL) UN/CREAT (RATIO) TLIPIDS (MG/DL) PLIPIDS (MG/DL) HDL (MG/DL) LDL (MG/DL) UA (MG/DL) AST/SGOT (IU/L) ALT/SGPT (IU/L) ALK PHOS (IU/L) GGT (UAL) SDH (IU/L) LDH (IU/L) CK (UL) AMYLASE (IU/L) LPICPOAASOE ((IIUU//LG)) CA (MG/DL) ION CA (MG/DL) IPHOS (MG/DL) NA (MMOL/L) K (MMOL/L) CL (MMOL/L) MG (MEQ/L or MG/DL) ZN (MG/L or PPM) FSER ((UMGG//DLL)or PPM)
7
002444
---
CovanIcMeT6T3e6292s-522s62
Abbreviations and Units for Clinical Chemistry (Continued)
Test Excess iron UTontbaoluinrodnibrionndbiinngdcianpgaccaiptaycity Percent iron saturation RPleadsbmlaocohdolcienllesctheorlaisne:esterase Brain cholinesterase Caudate putamen Hippocampus, Frontal cortex Cerebellum Bicarbonate Serum hemoglobin Serum bile acids Fecal bile acids Average fecal weight OFescmaollabliilteyacids (calculation) Electrophoresis Albumin Alpha-1-globulin Alpha-2-globulin Beta globulin Gamma globulin LHiogwh--ddeennssiittyy lliippoopprrootteeiinn InsuVleirny-low-density lipoprotein Adrenocorticotropic hormone Cortisol Glucagon Triiodothyronine Thyroxine CPraencarteianteick-isnpaesceifiiscoeanmzyylmaesse BB MB MM
AEbXbrFeEvi(aUtGi/oDnL()Units) TIBC (UG/DL) FUEIB%CS(AUTG/(D%L)) CHEP (MUML) CHER (MUML) CHEB (MU/ML) CAUD PUT (UMOL/G) HIPPOCAM (UMOL/G) FCECROERBTEELXL((UUMMOOLL//GG)) BICARB (MMOL/L) SER HGB (MG/DL) SFBBAA ((UUGM/OMLLI)L or MG/DL) FCC WGT (G) FBA (MG/Day) SMO (MOSM/KG) EALB (G/DL) EA-L(G/DL) EA-2 (G/DL) EBETA (G/DL) E GAMMA (G/DL) E-HDL (%) E-LDL (%) E-VLDL (%) INSULIN (UUML) ACTH (PG/ML) GCOLRUTCIASGOOLN(U(GP/GM/LM)L) T3 (NG/DL) TP4A(MUYG/LDL()UL) CK-BB (UL) CK-MB (ULL) CK-MM (ULL)
002445
38
-_--
CovanMcTTe.6a6322s90-25s262
Codes for Anatomical Pathology
DistributionofFindings Focal Diffuse Multifocal
MICROSCOPIC CODES
G1 rades for Severity oMriAnimmoauln-tthe least amountof change that can be observed with the
2
light microscope Slight - less than average amount of change, but readily discernible
3
as abnormal Moderate - the average amount ofchange that is expected foar
4
lesion Moderately severe (marked) - a marked amount of change with
5
pSoesvseirbel-e 2logsrseoatf afumnocutnitonooffcthhaenagfefewcittehdpcreollbsabolreolrogsasnsof function of
the affected organ
cell
or
organs
and
frequently
involves
large
areas
of
the
002446
39
|
mary a Sine,scion
5 +hor arm ee a8 =S
ea
PSSe
RL I WS re
ne
5288 @
"
228 &aS
SEARRT R S
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ee
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ores. 3, Eman,on cto
mans 1
238 aa
"
re TrRa
rum --
zz
ig
.
seasof snrvee.os
g88 BB B i BoH k aa. 8 4
H hoo Hi now. "
223
8aa
Id
macy of Rectal ety erat Sua (1
REE ERS Se ere
men
2 8a a
a
ww
moe xt om NWT aie NOW
82a8
ier our
288
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whee wie
reves or
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2
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8
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28228 *
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Sher SR Sa EN UR om om mu She WROD
88
a@2 5
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288 a>a
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a Th SEOVE
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322 daag
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8388 3
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2 a58 a
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ST SRnye Flow SMa EOE OM WmouE oS. mom
8883 32
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3 a58 3
n
8 3 3
mary of Cates maces ote
332 3I5]
hw Be ME Bu whe Se 288 2
Bho SE Sw ZEOVE 8% wR 2. mom
88 82
7s
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Shoo TUT VEDI EE Sa me mm our
2338 2
8 2 2
|
|
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hee oh Sw BOVE OM mom om Er
28 a@a
Ed
Sle CTE NTETOMTE WL ELS. mE ume eam
238 a
Shaan wi SES Be wha Sh
23: ?B3
0
Slee SR Sa EROUR Mw wm om EO
2 aI8] 3
5
||
Hare TET MET NATE mmm, me ume ro
238 a]|]
2
Shoe Se WR Ba whe
83 3a
8
macy of Clntens cmiotey oa
ET Te, PRT Sr seo
Shwe Sh Sa BEOVE SA um omy om Sh mn
83
R@Q 8
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ss
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8
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||
em S gyn es TtR weeg Se
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PfPiEi
88Q morioe @|]
tee or
---
Covance 6329-222 M3TMTe-6929s5.66
APPENDIX 1
Protocol Deviations Protocol
Protocol Amendment No. 1 Protocol Amendment No. 2 Protocol Amendment No. 3 Material Safety Data Sheet
92
02499
-_-- Protocol Deviations
CovanIcMeTT6-e36229s-0252s62
Protocol. Group Designations and Dosage Levels. Dose levels were 0, 0.02, and 2.0 mg/kg/day. The control group (Group 1) was to receive an equivalent amount of lactose in gelatin capsulaess the total material administered to Group 3. The total material was 20.0 mg/kg/dayforcach group; however, the low- and high-dose groups were to receive test material triturated with lactose (1:999, ww, and 1:9, ww, respectively).
Actual Procedure. Because the capsule broke during dose administration, the following
animals (Group
may not have received the full 1 male) on Day 1; Animal No.
amount 105367
of dose (Group
preparation: 1 female) on
Animal Days 2,
No. I05350 7, and 14;
Animal No. 105344 (Group 2 male) on Day 7; and Animal No. 105369 (Group 3 female)
onDay2.
Protocol. Observation of Animals. Rectal Body Temperatures. "Beginning 2 weeks before initiation of treatment, rectal body temperatures will be taken at approximately 11:00 on two days/week (Monday and Thursday).
Actual Procedure. Rectal Day -14 and between 08:17
body temperatures and 08:40 on Day
were -10.
recorded
between
12:50
and
13:30
on
Protocol. Serum PFOS Level Determination. Frequency. "Before initiation of treatment (Day -7or-6), andprior to treatment on Days 2 (approximately 24 hours after the first dose), 7, 14,and 29." Actual Procedure. On Day 2, incorrect blood collection tubes were used for the cleovlellecdteitoenromfinbaltoioodnsfowracsliunsiceadlfcohrecmliisnticrayltceshte;mtihsetrreyfotreest,s.bloRoedmaciolnliencgtesdefrourmsefrruomm PclFinOiScal chemistry tests was used as the Day 2 sample for serum PFOS level determinations. In `addition, whole blood (approximately 2 mL) was collected from a femoral vein of each animal before treatmeonnt Day 3 (approximately 24 hours after the second dose).
002500
9
-_-- Protocol Deviations (Continued)
CovanIcMeTT6-3e62w29-9s25262
Protocol. Additional Blood Collection. "At scheduled and unscheduled necropsics,
blood (as much as exsanguination."
possible,
up
to
20
mL)
will
be
collected
from
animals
at
the
time
of
Actual Procedure. Additional blood was not collected from Animal Nos. (Group 2 female), 105365 (Group 2 female), and 105369 (Group 3 female)
105364
Protocol. Postmortem Procedures. Tissue Preservation. "The following tissues (when
present) from each animal willbe preserved in 10% otherwise specified, for possible future microscopic
neutral-buffered examination:"
formalin,
unless
A(cGtruoaulp P1rmoacleedsu),re1.05F3e4m4oraanld 1b0o5n3e4m8a(rGrroowupfr2ommaAlensi)m,a1l05N3o4s5. 1(0G5r3o49u3panmdal1e0)5,351005366
(weGrreounpot1efxeammalien)e,damnidcr1o0s5c3o6p4ic(aGllryo.upT2hifsemtiaslseu)e wiserleistiendsuwfiftihcietnhte;atphperroepfroiraet,etcheosmemteinsstuses
in as
the pathology data sheets having been examined.
for each
individual
animal.
Summary
tables
do
not
include
it
These deviations are not expected to have affected the results of the study.
02501
%
_--
Protocol
lof 18
Counce 6320222 mews
C02502
95
Covance 6329.22
-_--
Tem
2
9%
ee
002503
Covance 329222
_--
Tews
3
002504
97
Covance 6329222
_--
mews
4
002505
98
_ 5
WirCoveanIcMwe 6T2s699.25s262
002506
9%
Covance 632922
_--
MTews
6
002507
100
Covance 6329222
_--
rem
7
002508
101
-_
SiCT ovane 3cMeT6-w36229-0s25262
8
002509 102
-_--
Covawn3ciMeTr6-362e29-0w2526s2
9
02510 103
-_
SiCTovaneIcMeT6w -36292-s 025262
10
02511 104
-_
00 CovaNn3cTIeM6Te3-269w2-02s5262
u
02512 105
_ 12
CovaSnIcNMeiT6.3T62e92-9w252e62
106
002513
TCN ovanIcMS eT36292.925262 13
0251.2 107
-
14
mCoe vTeM 6Tm3.2092.92526
002515
108
-_--
CovanS3cMeiT6-r3622e00-5w2.26s2
15
00251 109
_--
CovaNn3cTMeT6e-36292w-925s262
16
110
002517
-_--
TCRovaTnIcMeeT6-362w290-52s.262
17
C0218 11
-_--
CovanIcMeTe6-362v92-02e5262
18
02519 12
-_--
Protocol Amendment No. 1 Page 10f2
Covance 6329-222 MTIeMTw-6s29s56
13
02520
Page 20f2
0t CovanMceTe 623200-52262
14
002521
_--
Protocol Amendment No. 2
lof2
Cove 632922 ures
us
002522
-_--
Covan3cMeYT6-362e92-025s262
20f2
116
02523
Protocol Amendment No. 3 lof3
CovanMceT3e299-52262
i
002524
_--
0 CovanIcMeST662e92.9m25262
2
002525 18
-_
0TCS ovane IcMeT6s -36292.s 025262
3
119
002526
-_ Material Safety Data Sheet Page 10f6
ee CovanIcMee T6-36220-925262
120
0025227
--- Page 20f6
0 CovEanMceeT6e3e2290.52262
121
002528
_-- Page 3 of 6
Covan3cMeiT6-362e299-52s.262
122
002529
-_ Page dof 6
00000 R CovanIcMeTR 6-36229-025262
123
CO:2530
_--
Page 5 of6
e[ wee-- rs
1
002531
-_--
Page 6 of 6
Covance 6329-220. wrewss
125
002532
_--
wirCoveanMcwTe 663s2209-s52262
APPENDIX 2
Individual Animal Fate Data Individual Clinical Observations Individual Rectal Body Temperature Data (C)
126
002533
2283 a4
RRLRESe
ol
a833
&
os
oH ots ore iosraison srone Kio
ner. 2
88 a&a
129
I ROTA EE eres v1 ERS Src acto
nr: 3
2283 aa3
130
3 8a88
Bl
gS 3a8
m2
| 228 aaS 153
g
RB
134
iit caioyneer 5
en
2&8g
8
1s
288
28
16
_--
MrCoavanMwcTe .6s36290.s52262
APPENDIX 3
Individual Body Weight Data (kg)
Low
02544
aa cesen enpoe sre cto
[SN
82S
aaa
138
via soe vei saa ha
8223 saaq
139
-_--
Covance 6329-222 M 3M TT -629s 5.6
APPENDIX 4
Individual Clinical Hematology Data Individual Clinical Chemistry Data
.
140
02547
wie ae me we an VOW Wome mmm wm Or 3 rae eels 0. naseti meray
832 3aa
ro: 3 onetevets 8 onagetits morgrtny
[I settee ferme eee
822
58
0
wine yr
Gon 3 oete 003 ms ek 50
una tics marin an
38
a
ws
Geet 00 onts rey 832 aaa
In
|
.
|
jo 1 Done Leva 0
`Dosage Unit: manasdny
Grow: 3 Dose Levels 0.02 DosageUnit: maskadey
et3 Dose evel: 2.0
Dosage Unit: mararday
a883
8a
us
han 1:74
Co
:
:
:
:
:
'
ies mn
882
a
16
tTRt nemensom nm s W ae hy - xe
Ee
Group: 2 Dose Levels 0.02 Dosage tic: maskadey
:
:
:
:
Group: 3 Dose Levels 2.0 `DosageUnit: markarday
:
a88
ai
wo
Ss [A --
et il 08
i at
Cod tn 20
sr
g28
ee
148
To
ee
oo
oo
}
rope 3 ova tev 6 [a --
Go3 tose eral 10 sae ics mney
823
gaaa
149
man ate on VO WOROT MN. TC mh Gro 2 so tov 0.03 age niesany
822 Jaa
150
rir mm
Group: 1 Dose Level:
DosageUnit: sakasday J
Grow: 2 DoseLevel: 0.02 Dosagevast: sarmasdey
:
:
:
!
:
!
Grow: 3 Dose Level: 2.0 Dosage Unie: marg/dny
!
8882
aa
51
reves or
}
Gro tone eve
--
.
Grom 3 tore tev 0.67 onsets monartny
003 Dots evel 20 buen wits soar
232
aa
152
rr
}
Groups 1 Dove Level:
Dosage tac: ma/rasdey
EE
ows tas 28 mae mts mr
28&8
82
15
!
et mens imi
Grow: 2 Dose Levels 0.02 Dosage Unit: mg/kg/day
:
:
388
82
156
i
Go ees a ms an
m3 net 052 ne is rr
rm es 0 vantis mi
388
8
15s
Grow: 1 Dose Level: 0
Dosage Unit: mg/kg/day
rs 01 owns carat
Grow: 3 Dose Level: 2.0 Dosage Unit: markasday
3888 a8
15
So
!
!
!
:
ST Se
Group: 1 Dose Levels
Dosageunit: saskasaey
Co
ns tris 20 ove is min
&822
2
15
et toi 6 oe es ren
G3 na 20 oe mis sare
88a
a
15
mo
wie ra
wt eee im
ns res 0 eaetis sat
a883
8
1
a SE
Grow: 1 DoseLevel: 0
`Dosage Ueit: markardey
Cot rom 00 en mis mer
Sa
oT
Grow: 3 Dose Levels 2.0.
`Dosage Unie: sakasday
!
:
!
:
8822
28
1
!
Gro 1 toeva0 sae niesmain
Grom 3 tose eal 0.03 ons nis morartny
mee veh 20 one tts manny
2233 aa3 161
re rs
Grou3: DoseLevel: 0
DosageShit: sarkasday
Sr
Grow: 3 DoseLevel: 2.0 Dosage Unit: mariarany
388a
2
:
;
|
i
Group: 1 Dose Level:
Dosage test: manasa
or
nt es 1 ove i arr
3 tos 20 ae ts morn
8
&3
@
or 3 oe es 2 ae issaat oo mans 20 nis mtn
83
4
ot
Group: 3 DoseLevel: 2.0 Dosage Unit: moska/day
:
;
!
!
'
822
N
6s
28 a&
166
[i
3 res 1 ne is ain om1 tis 20 ns ar 8 &34 0
Grom 3 Dose eve 0.02 Deen ies mbar
322 a
a
168
| |
3 tos1.2 sise rey
'
8 a34
i
wot tes8 an ismri
3 we 30 sn is sir 8 &3 mn
Grom: 1 Dose Level:
Dosage Lait: markgrday
Co
Grow: 2 bose Levels 0.02 Dosage Unit: markadey
!
3 tas 0 ove is i
a83
3
m
Grom 3 tosetowels 10 ons cits sahara 328 ae3s m
|
ee
RE
0m 3 Dose wks 20 nen nts maha 832 32a
ms
C3 Doanberks 002 menetits maar 822 2a
ms
[A
et meme in
Grom: 2 Dose Level: 0.02 Dosage Gait: makasday !
:
:
:
:
Group: 3 Dose Lave: 2.0 DosageUnit: merkorday
:
'
888
a8
ns
238 a@&
6
meme smi
Grow3: oseLevel: 2.0 Dosage Unit: mg/kg/dny
'
a228 28
m
Grow: 2 Dose Level: 0.02 Dosage Units morkarday
:
:
!
!
3288
8&
mm
28
19
238
&J
150
|
ot sre
|
Grow: 1 Dose Levels 0
Dosage Unit: me/kaiday To
Len .23522
pd
|
rer
n> tes2 sise meer
38a
8
wn
i
A, onan ties manasiny
G00: 3 Duttev 0.08 oan nies sry
38 Sa@s 183
}
Grom 1 omen tow 6 fp
Grom 3 tose tev 0.03 congenic: rnp
822 2@a
184
APPENDIX Individual Animal Pathology Data
Covance 6329-222 IMT-62056
oss
002592
238
a
186
|
mn apces sren cna sre eso
noe 3
882
%a4
157
228
a&
158
2 3
189.
E8aS IN]Y 190
|
ps
em
|
op ce i.
32.22i
om arms once sr rece susan sco
noe. +
832
3Ia]
192
= EEpt a
EE,
832
8Q
193
82
8BR
198
888
8
195
I
|
omgr psse otsn se sro
ns
82888 196
3
88
2
10
_--
mrCoeunIcmMeT6-3s6229-0s25262
APPENDIX 6
`Summary and Individual Hormone Analyses Data Figure 1 - Mean Estradiol Data - Males Figure 2 - Mean Estradiol Data - Females
Figure 3 - Mean Triiodothyronine Data - Males Figure 4 - Mean Triiodothyronine Data - Females
NOTE: This appendix of the report contains information supplied by Ani Lytic Inc. and has been reviewed by the Quality Assurance Unit of Ani Lytics Inc.
~ 198 --
CORS05
Grow: 2 Dose Level: 0.62 Dosage Duk: markarany
:
es it on i
8882
32
1
[RA
ne
SETHE TET Tne megs oman
oes io 0 i Gre onton 30 tse nes sarin
88 38g
0
mRet CeREnRE TR aTi ORR meee
oes ei 10 i
388
28
on
and Eedividunt lorsone Noises uta
es ir eC
ow
Ed
rd Owe eels 20 mua niesary
i
222
g
=
Figure | `GRAPH- Mean Estradiol Dita - Miles:
228a23 13
20
Figue2 `GRAPH- Mean EstrDaatad- iFemoalles
38
a8
=
EY
Figure `GRA-MPeaHn Triodothyronine Da-Mtalaes
28882
25
Figured `GRA-MPeaHn Triodothyronine Da-Fetmalaes
228a23
26
--_--
MCovranMecTew-6362s209-2s5262
APPENDIX 7
Dose Analysis
NOTE: This appendix of the report contains information supplied by the Sponsor and has been reviewed bythe Quality Assurance Unit of 3M.
02614
207
-_--
Covance 6329-222 M3TMeT-M62s9566
Dose Analysis (to be provided by the Sponsor)
002615 208
-_--
CMovranMTceTe6e3w209-5s226s2
APPENDIX 8
Serum and Liver PFOS Level Determinations NOTE: This appendix of the reportcontainsinformation supplied by the Sponsor
and has been reviewed by the Quality Assurance Unit of 3M.
002616
209
_--
CovanWIcMeiT6r-362e92-s925262
Serum and Liver PFOS Level Determinations
(to be provided by the Sponsor)
002617
210
_--
CovaMnIcrMeTT6-3e6229w-02s526s2
APPENDIX 9
Cell Proliferation Evaluation
NOTE: This appendix of the report contains information supplied by Pathology
Associates International of Pathology Associates
and has been International.
reviewed
by
the
Quality
Assurance
Unit
002518
21
-_--
CovanMcreT62e3290s5.26s2
Cell Proliferation Evaluation (to be provided by Pathology Associates International)
02619
212
(PZ?)
A
Pathology Company of
AScsCisoeornpccoieraaAtptepisloincIanttieornnsaItnitoenranlational
=F EpOmeesy.
CELL PROLIDFERRAAFTTION REPORT
+-WEEK CPAOPTSAUSLSEIUTMOXSIACLITTY(PSFTOUS;DYT-W62I9T5)HTIPN0ECR2FY9LNUO5OMR.OOLOGCUTSANMEONSSUALFtOSNIC ACID
COVANCE STUDY NUMBER 6329-222
PREPARED FOR: BUILTDOINXGIC2O20L-O23GE-mY02S, E3RMVICCEESNTER
ST. PAUL, MN 55144-1000
PREPARED BY:
PAT1H5 OWLOORGMFYRAENAD'SESSROIMCCIIKLA,LTEMCSDOUIR2N1TT7,E61RSNUAITTIEOTNAL
SEPTEMBER 30, 1998
TS Warmes Ml Cour, ute 1+ Frederic, Warrant STOTT GO SIT
002520
`CovDarnacfet CSetluldyPrNoluimfberearti6on32Re9p.o2r2t Page?
CELL PROLIFERATION REPORT SUHLFVFOENIKC CAACPISDUPLOETTAOSXSIICUIMTMYSOASNLTKEUEDEYROWSI;TTH62P9E5R)FILNUOCRYONOOCMTOALNGEUS
COVANCE STUDY NUMBER 6329-222 PURPOSE
mIoahsteeerapinaulrdpoolnsoewcreiortfidcatolhseeenslztevuyedmlyes twleoavsbeles1,u0shepodrromivoniancdeehsr,doaantnaidcfoottrohxedirectsieteylremscittnueiddnybgaioanncdh0eesmtiaicsmasaeltspesdatrhmaeamxeftieamerutsm-otoaleratsed (CpTPhraiRseOgSdreep,oprctah,veeseuckbe.mliCtpatropelsdiuflbeeyratPTioaoxtnihcofiiltnoydgiySngtAsusdsayoncdiwaiitntethserPpIrenerttfeamltaiuotrniooofncoatrlaCn(oePvAaSDnu)lcfetoonSittchuedAysctiNuddoyPmSaoproons6ro3rs5,9-30M3,5 Rkeagbuiolrsaat:tiosoTrtn6yus2d,9yP3rPa)awcteiineceCcy(onGnodLmuPoc)tlegRduesgiunMloacntokimoepnylssi"aa.nsceAsltwliatfshopretthchet.sinEofntTvhiilereotn4ams0ek.nsotaafslstohPcerioatUteSec.dtiwoCinotdhegpseinogefs."pForcrtro2io2onnn0 applicable amendrmte7n9t2s,.issued November 29, 1983 (effective December 29, 1983), ang wiiany
MATERIALS AND METHODS
Tissue Collection for Cell Proliferation
iraTnenwpdonroeaensneeinamtnaailtmsiapvleepresrsaesmxepxilneoicnfontdtohrseoellgigvrreorou,uppte3s1t,(es2t.h(0rmemaelgeaksngiiomdnalalysy))paewnredrspeeaxsnacicnrreidafosisceferdoagmrnoeeusapcf2hou(ar0n.iw0m2aalmegw/ekEgZ/fdooatcy))
pbSrploeolcciikff,eicraa2ttiifonoSnglrsim.cdaeell,liTnnwi,usacssupreopbcrleeospcsakersdedweafrnoedr tehHem&nbEesdhidepevpdaelduatttooioPpnAarIaaffnfodirnsiebmclmtouicnoknoihnibgsytaoncCdhoesvmtaiancicaneli.n"g"pdeeFrsrpaarcmottoeiccnogtl)i
lear antigen (PCNA), amarker of cell proliferation.
Immunohistochemistry for Cell Proliferation
S{faaocpnteairortnrsosoStfanpPdalurasar,fdfFiiinms-mheuemnrboeShdcidiseetndtoitcfihisecs,muiePsciatwltesrmbeuertcghuhot,dasPA5w)eurmteo Standa
aennsdurpelacaeddheosniopnosdiutriivenlgy used to stain issues for
pcrhoacregsesdinsglifdoers BONA (GALL
mrSeeecttahigoondersndfwoterOsrpteehrreeiadqnteucitiunerbgceatdtPefrdoorcwetidhtuehraaevimfdooinrno-cIbilmoomtnuiannlopkaeinrstotixobicodhdaeysmetio(cPaAlCBNCStAaKii(tn,iDnAlgoKt(O#S,POKlPo-t5##170001706),..PPAABTIriNeof,ly,AK1t74i2s3s30u))e
soeycttihoenschwreormeacgoeunnte3r,si3to"andioinfaemdtihwneiotabhnethniezgmieadnti-onaxenytli(ibDnoA.dBy;coSmipglmexa.CPheCmNicAalexGproe.ssilootn #in1c8eHllSs2w0a%e,eaossee)
Cell Proliferation Measurements
s{3Tte0haa0seit0npi2ehn0re0cprea0nttoaaccgiynetaeorsfciepnlrlo1sl0iofffeirtealhtdeisnpgoafnccellrilevsears(,paproelrilfeaeansritamta5iln0g.0iLnAedyenxde,iggaPtIci)evlewsacsoofntdtreohtlertsmeiisdnteesdw(mabasylesiscnocariniaind)ge,datiolsreatsst
g un and consistedof study tsse that was not incubated with the primary antipeds,
002621
CovDarnacfet SCetluldyPrNoluimfberearti6on32R.ep2o2r2t Page3
fhlFoioirsrttlociyemvlolroppfrhootllaiofigenriiacntgic,ohnapnergvoeacsleu.sastiiCnoenglsl,apnrsdolliidsfeeescrtawiteoirnoeninwfgi,arsstptohpteeennrqtuuisaaelndtpaiatftilteoedmwsamtaofghniigcfehileclraultmaiarognnpir(fo1ifc0fa0ot)iaotnitoo(rju<d5g;e
Iusssseedr oayndc4ya0l0uXatifnogr ttehseteHs a&nEd evaluated for cell proliferation.
psaindcereparse)paasreddescrfibredom
thaebosvaem.e
Htiissstuoemoblrapchkeifonuy ewaochr
122e0r0sX corey
Statistical Analysis
Due to the small sample size, statistical analysis was not performed. RESULTS
Cell Proliferation
tihnedilivviedru,alteasntiesmaalndcepllanpcrroleiafseorfaticoonndtartolaaarned ptersetsmeantteerdiailn-tSreecattieodnaInTim(aTlasblweas1).simCiellalrproliferation in Histopathology
tiShetechtiimhomenumsnafotrosxotymaliitnhneedasnsadlimdceeoss.tiinsIsnu(deHiv&bilEdo)ucaklfsoarunsihemidastlfoopfraitnphdroielnopggsairacarteeivopanrleuosafetniPtoCenNdAtio-nsfAtapacpiiennlededintshl1eidiesntweerrperesttaained tToSiufisgns/vguklegee/sdsaeaycn)td.iopanansndocfrhleiiagvshe-rfd,ropsoaemnc26r.e0yaosmi,igna/gnkdga/dtduealstyte)fstefrmreaoalmtem6emnyotongukrneogyuspadsruelwptermreaeselenevtiamnlogunaktcoeendytsrhoials,ntldloogsiwiendgaltoelcyse.ec(tBoia.o0nc3s ctoenrfmoiunnedecttiwhohenestwpheaecsirfoiscrtiantioynteodmfowtrhipethhPoClhoeNgmAiactsoctxhayialnniignnegsaonwbdesreeerovsoeicdnc.uirnrtTihhneegseipnautnrhipemosaslees.toisfsuteisswehviaclhuamtiaoyn awlaess otro `tTwehshetiiccrhuelwsauorluttlsidsssauhleotsewreftdrhoemtihnamttaerlnpeoremstoiagtnnikiofneioycfsantthorecPhtaChneNgAelisveswrteaarinnedinogpbasinnecrtrvheieasdsstiutndisyes.iutehser ftrhoemlifveora, lpsan`cmroenakse,yso,r DISCUSSION
i(mInn0od.nst0kh2aeemypgsr/,eksgea/nndtdasytt)hu.edyal,invdcerhliagpnhrdolpiadfonescreea(te2iao.sn0owmfagfs/eKmgma/eldaeasyum)roegndrkoewuiyptsshifanrfteohrm flcoiouvnertrr,woletees(kt0sesmogan/nkdsgt/pudadanyyc)1r,eadslootowufrdmaoamseleee bnidoltoogmiacasatsleepsdasrmtaamhXetieemfrfusenc,ottlooeftirhnacetleutddeestdcomealslteeprraionaldlifoleronawtceirrointd.icoasleenlezvyelmsetloevbeelsu,sheodrimnonaecsh,roannidc otodxeicritsyesatusd)y pTahnecrreeadsi,dansodteatpepremairnetdobbyeparocleilfeprraotliinfgeirantdiivceesrewshpiocnhsewetroetshiemitleastr imnataellriaanlimianlst.he liver, testes or SUMMARY
iinnctrheeasperdesienntthsetuldivye,r,cetelstpersoloirfepraantciorne,asasofdemtoenrkmeiynse.d by measuring the proliferating index, was not
002522
CovDarnacfet SCetluldyPrNoluimfberearti6on32R9e-p2o2r2t IL TABLE
002623
`TABLEC1O.CVEALNLCPE RSTOUDLYINFO.E6I3R2N9AM-O2T2N2IKEOYNS
en [EEE p
Animal Number
[11--00mmog/tkagi/doaayy C(oConntiroal [|mT|tioosssiss0||
2-002 mofa/day 2-002 mafoiday
Low dose) |__|losses| (Low dose) | M_|105347|
[2---02002mamkaaktalidaayy(ia(Lnowdodsosee))||_u_||l1o0s5s0e45s|
[i1=--00mmootkggiiadsayy (CCoannttrool)
Tr T+
"108565 | |" 1osser|
[[22--00022mmaafngaiioonayy
((LLoowwddoosses)T||+ + |
105364 T tosses |
92--02002mofkmgalfdaaidyay (Hi(ghLowdodsoes)e)||F 'F_[|"i1o0s5s376o9||
Index Liver.
index Testes
Index. Pancreas
ozs 0.087% |
ssopmoneT|amsomaoen ]|
0.056%| ie.sw 0.00% | du.s%
| esaon | |"sssow |
0.106% | 0.026 |
1t6r.e5%%--T|esesea0oen--|]
0.000%| 0.000 |
tia ha
samen--] saser--]
0.000% 0.003 |
Na ha
ener] saeer--]
0.146% | 0.000% |
wa nA
| |
eozow | socom]
02524
} `CovDarnactet SCtluldyPrNoluimfberearti6on32R9e-p2o2r2t
APPENDIX I
002625
`CoStudvyNuambenr 6c329.e222
Individual Anidm-awleeHkistCoappastuhloeloTgoyxiFciintdyinSgtsud-yMale Monkeys
|E NumE ber|
| S|] Paar --
Individual Animad-lweHeikstoCpaaptshuolleogTyoxFiicnidtiyngsSt-udFyemale Monkeys
rem os Number
oS
oss
Tor
J)
Toor
TNosignificant findings | Nosinificantfindings
|Normz----------,
|Nosignificant findings] [No sianificancfincings----|
00R5R6
Cormaet CSetyPotttenienrgr
III. SIGNATURE PAGE
4-Week Capsule Toxicity Study with Salt (PFOS; T-6295) in Cynomolgus
Perfluorooctane Sulfonic Monkeys (Covance Study
Acid Potassium Numb
222)
er 6329-
`Submitted by:
PAIProject Manager:
cer THE] Sandra R. Eldridge, Ph.D.
Loop
Date
PAI ProjectPathologist: Carolyn Moyer, D.V.M,, Diplomate, A.C.V.P.
nei Date
CORGR7
. `DCraftSoCteuldvyPrNoluaimfberenarti6o3c n29R-e2pe2o2r.t
IV. QUALITY ASSURANCE STATEMENT
02628
DO]
.
ala 4
PAatChooloomgfSycp iAensceasApoplcniciatay iotn eIsnteIrmnattoenerlnCoarptoiraotinonal
She fc]
Cell Proliferation Report A4c-iWdeePkotCaaspssiuulme STaolxtic(iPtFyOS;udyT,-6W2i9t5h) iPnerCfylnuoormooolcgtuanseMoSnulkfeoynisc
Covance Study Number 6329-222
QUALITY ASSURANCE STATEMENT
f2{Pe8arbooleoesitoMrnedecAeorgnradctmyoaof.nCthoTeephrnedosLjppaeaobttcohihroiaalostonogbsryeyareuPeniptaIosnrcstpiseca(tnGedLacPac)unrreaagtueluartotifooncsbtyirotonhmoufPgtAaIeiGssudlebtiyyshAesseucr2.anEcesrUfanoill(oAAwYiUn))g performed and repareg by the Cc
puteato oof Inspection Q086//2274//9988,09108,10/98 08/27/98i08/08,10/98
Phase Inspected DSLatrbuaedlyGiDenagltaPraonfdorRaeiloanteFdapDooycsumentation
DatetoF_iMnadinnaggsemReenpto/rted Study Analyst 0086//2276//9988,00/10/98 08/27/38;08/10/08
Ree QO Budasp QualtyRoAssseuAr.aBnicsehoApuditor
DegnanienTe1000 Dee
0R629
3M ENVIRC DNMENTAL LABORA' [ORY
Protocol - Analytical Study
Quantitative Analysis
(PFOS, T-6295) in Cyno
of Perfluorooctane Sulfonic
molgus Monkeys Following
Acid PotassiumSalt
Administration ofa
4-
`Week Capsule Toxicity Study
3MET & SS Study Number:AMDT-041598.1
In-Vivo Study Reference Number: Covance 6329-222
`Test Substance: Perfluorooctane Sulfonic Acid Potassium Salt
Sulfonate) C,F,;SO,K"* (POS, T-6295)
(PotassiumPerfluorooctane
Name and Addressof Sponsor:
3M Toxicology Services
Building 220-2E-02, 3M Center St. Paul, MN 55144-1000
Name and AddressofBiological-PhaseFacility:
Covance Laboratories Inc. P.O. Box 7545 Madison, WI 53707-7545
Name and Addressof Analytical TestingFacility: 3M Environmental Technology and Services 935 Bush Avenue St. Paul, MN 55106
Dateof Approval by Sponsor:
Proposed Analytical Initiation Date:
Proposed Analytical Completion Date:
4121198
7/15/98
7/15/99
Author: Glenn Langenburg
Study DireCi
#
fate
ford 4
Principal Analytical Investigator
@ LBiolsical Firs Sr r
efor
ee
0630
3/5/55
Date
,
10 Purpose TmehtiasbsoltiutdeysisindecsyingonmeodltgouqsuamnotnitkaetyisveolryadlelyteardmmiinneisttheereexdtePnFtOoSf.absorption ofPROS and varioms 42T.h00iCsRFsEtRuGdpUyaLrtwAil7Tl9O2bReaYncCodnoadMpupPclLtiIecadAbNilnCeaEccucrorerndtanEcneviwriotnhmtehnetaElPTAesGtoiongd&LaSbaofreattyory Practice regulations pSrOoPto'csola)n.d methods (see sections 8.0 and 9.0for specific SOP's andmethoSdersvirecleast(edEtTo&SthSi)s
3.0 TesTMATERIALS
3.1 T3e.s1t1, CoAnntarloylt,icaanldRReeffeerreenncceeSSuubbssttanacnec:esPearnfdluMoartoroicctaense Sulfonic Acid Potassium Salt
3.1.2
(GFpSOK" (PROS, Analytical Reference
T-6295)) Substance
Matrix:
Cynomolgus
monkey
serum
and
liver
tissue
3.1.3 Atinsasluyet(iCcoanltrCoolnttirsoslueS,u1b0s5t34a6n)c.e Matrix: Cynomolgus monkey serum and liver
32 Sourceof Materials
32.1 322
AAnnaallyyttiiccaallRReeffeerreenncceeSSuubbssttaannccee:Ma3trMixI:CPC/oPvCaPncDievRiessieonarch Products (The
323 sAonuarlcyetoifcatlhCeomnotnrkoleySsubwsiltlabnecedoMcautmriexn:teCdovinanthceerRaewsedaartac)h Products. The tissue
was from a control animal, 105346.
33 N33u.m1berTeosftTseasmtplaensd Control Samples
Number
Dose Level
&
PFOS
T 3SMeaxle | Onlly by (mg/kg/day)
oe EE 7
Vale | Onllyby
E Em]EEErE E] 3
Orally by
Collections!
= Blood predose (6 days fom dosing), and
_ =
BLilvoeordtipsrseudeosoen(D6ayda2y9s. From dosing), and
+_ =
BLlivoeordipsrseudeosoen(D6ayda2y9s from dosing), and
Bd coToml sie sWI TyCova, eri i+e odLeivseriSsFsuRe oBn DTay 29S.STor 34 ICdoevnatnifciecaattitohneotfimTeeosftcaonlldecCtoinotnr.ol Samples. Test samples will be identified by
02631
2
35 rPeusrpiotnysiabinlditIyodefnttihteySopfonMsaotre.rials. Characterizationofthe test materials isthe
36 SctoanbdiiltiitoynsofofRaedfmeirneinstcreaMtaitoenriisatlh.e TreessptoinnsgibfiolrittyhoefsttahbieliStpyonosfotrh,e test materialunderthe
3.7 aSmtboireangtetCeomnpdeirtaituornes.foArnaTleystticMaaltererfiearlesn.ceAnmaaltyrtiicceaslwriellfebreenscteorseudbsattaonrcebewlilolwbe stored at
approximately analyzed. Test
-20C, until and Control
esxatmrapclteisonw.ilAlbllerexetcreaicvtsedwialclcboerdsitnogretdo
nominally at 4C AMDT-S-10,
until
38 tDiimsepopseirtiioodnaosfpSepreGciLmPenrseg.ulBaitoiloong4ic0alCFtiRsspuaersta7n9d2.fl1u9i5dsfwoirlsltubdeiersetlaoinngeedrftorhatnh28redqauyisr.ed 39 aSsasfoectiyatPerdecsaauftetiyonisn.foRremfaetirotno. tEhxeeMractiesreicaaluStaifoentwyhDeantahaSnhdeleitnsgfkornitvheessfuobrstcaunttciensgutsheed for
sshaomuplldeamlawtraiycsesb.e wSoafrentywhgleanssweso,rkpriontgecitnitvheecElnotvhiirnognmaenndtaaplprLoapbroiraattoerhy.and protection
A4.n0imEaXlPsEwReIrMeENadTmAiLnis-tOerveedrgveilecwapsules orally, daily (7 days/week) for 28 days. Dose levels oF
thPiiFsgsOhu-Sedsoiasnmetphrleeescspaewpcsetuirvleeelcyvo)al.rlieeBcdtleo(d0oa,dtt0s.ha0e2m,ptlaienmsdeow2fe.0rsaemcgcroi/lfkligec)cetb(eedDtaawtyere2e9n)g.uglraSoreuirpnastea(rcnvodanltlsriov(les,reeltio3sw.s-3u.d1o)s.e,Liavnedr
2s2hi2pfpoerditnof3orMmaEtiToAnSrSegaforrditnhge tahnealiynt-ilciafle ppoorrttiioonnoofftthhiesssttuuddyy..
See
Covance
e were Study #CHW 6329-
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`metabolites.
luorochemical
L5i.v0erEXtiPsEsRueIaMnEdNTseArLa-wiAlnlalbyetitcaarlgetMeedtfhoordansalysis. Methods are available for extraction and sTaanhamelpylmseeitsahnooafldfyslsiuwsoi.rllocbheemvialciadlastefdrofmorsceyranoamnodllgiuvesrmtoisnskueeyvamraitoruiscaensipmrailormatotroircecson(csuerereantttalcyhwmietnhts).
SoDmadwawyes
0000000000000
cDoantcaenwtilrlatbieonroefpofrltueodriadsetpheermtiilslsiuge,raomrsin(mogt)heorfcPleFaOrSly dorefmienteadboulniittse,paesraupnpirtoopfritaitses.ue,Quoarntasitaation
3
02532
rEweyilgelrreabsgeseispoeanrnafdnoarslymtsaeindsd.uasridStndagteivcsiatlaiitcbisrouantssieoodfn, cacuorntvcheesn.dtirTsacthrieeostneisocnfouorrfvtdeihsfefweiPrlrelinntbcediopdaelteInrvmeisnteidgautsori,ngmlaiyneianrclude.
simple
statistical
tests
such
asStudent's
test
may
be
applied
to
se groups. Ifneceseary, determine statistical diffsence.
NinoclfuisniaolnrienpotrhteiwrilflinbalerperpeopratrfeodrfCorHtWhs6s3t2u9d.y2.22A.data package will be provided to Covance for
7.0 MAINTENANCE OF RAW DATA AND RECORDS
7-1 TachceorfdoilnlgowtionAgMrDaTw-dSa-ta8.and records will be retained in the study folderinthe archives
7.11 7.12
AStpupdryovceodmepsrpootoncdoelnacned amendments
7.13 Shipping records
7.14 Raw data
7.15 7.16
Approved Electronic
finalreport (original copiesofdata
signed
copy)
72
1S0upApMoDrtTi-nSg-r8ecwoirldls itnoclbueder,etbauitnendotsenpeacreastsealryilfyrobme
the study folder in the archives limited to, the following:
according
7.2.1 7.2.2
Training records Calibration records
7.2.3 7:24
SItnasntdraurmdenOtpmearianttinegnaPnrcoeceldougrses, Equipment Procedures, and Methods
7.2.5 Appropriate specimens
8.0 REFERENCES
8.1 82
AMDT-S-1, AMDT-S-2,
Chemical Solutions
Tracking and Standards
Making
83 84
AMDT-S-3, Personnel Training AMDT-S-4, General Lab Documentation Systems
85 86
AMAMDDTT--SS--65,, GGeLnPe-rraellaLtaebd RDeoccourmdesntation Systems
87 88
AMDT-S-7, AMDT-S-8,
GLP-related Archives
Records
89 8.10
AAMMDDTT--SS-.190,,GSLaPmpPlreogTrraacmkainndg RSeyssptoensmibilities
8.11 8.12
AAMMDDTT--SS--1152,, LAanbaloyrtaitcoarlyMPeratchtoidceVsaalniddaDtaitona Management
813 8.14
AAMMDDTT--S$--1167,, QAunaalliyttyicAaslsuErqaunicpemeUnntitSystems Documentation
002633 4
8.15 AMDT-S-18, Routine Calibration Checksof Balances
8.16 AMDT-S-20, Daily Calibration Checksof Automatic Pipettors 8.17 AMDT-S-21, Annual Calibration Checksof Automatic Pipettors
9.0 ATTACHMENTS
9.1
9:2
FUFsAAiCCnTTg--AMMn--a21l..y0s0iAsEnxUatlsryaicsntigisooHnfPoLFflCPuFoEOrlSoecchtoerromsiOpctrahaleysrMiAannsiLsoivnSeiprceEcFxtltrurooamrceottcsrhuyesmiincgalHPSuLrCfacEtlaencttsrforspormaLyi/vMeasrs
9:3 FSpAeCcTt-rMom-e3t.r0y Extractionof PFOS or Other Anionic Fluotochemical Surfactants from Serum
Using Analysis Using HPLC Electrospray Mass Spectrometry
9.4 SFpAeCcTt-roMm-e4t.r0y AnalysisofFluorochemicals in Serum Extracts using HPLCElectrospray/Mass
--
002634 5
3M MEDICAL DEPARTMENT TOXICOLOGY SERVICES EOcc:TJEohE Nn SBA EuRtenThoff From: Marv Case Subject: Covance Study 6329-222 Date: 27 July 1998 On 22 July 1998 when I was at Covance Madison, I reviewed the microscopic liver tissue sectionsofthe monkeys on the 4-week oral rangefinder study of PFOS (Covance study 6329-222). There was no evidenceofcompound-elated effects in the liver even in the High dose animals. Our toxicology consultants had asked the question whetherthe high dose monkeys had any evidence ofliver necrosis or any other microscopic evidence adverse liver effects. The answer is negative.
C02635
