Document jNdbo9exoLM0n8aekq7GZG6Z
~
. 8EHQ-- 0800- 375
AR226-0960 FA
ori ed ADRAFT 5/1495 7
3M MEDICAL DEPARTMENT, TOXICOLOGY SERVICES
!
Report for Study No. T-6316.9; DT21
ORIGINAL
In-Life Start Date: July 8, 1998 In-Life End Date: July 28, 1998
`Background:
<ontalis Xo cpr
In an attempt to determinethe sourceof low-level perfluorooctanesulfonate (PFOS) body
burden found in control rats involved in some 3M contract dietary studies, a
comprehensive plan with the following objectives and responsibilities was designed:
Ohobujseicntgairv1eea-oTfothienvceusrtriegnattedpioetteanrtyisatlusdoiuesrcoensopfecrofnlutoarmiinnataetdiotenstwictohmipnotuhnedsstuadty
Covance Madison. Dr. Andrew Seacat was appointed study director with Dr.
Marv Case as alternate. Jim Wolters, 3M Environmental, i responsible for
coordinating and conducting direct air monitoring and wipe samples.
J Objectiv2e - To determineif contaminationoffeed is leading to the low levels of
"ZFiv
PFOS seenin control rats in the two-year dietary study of N-Ethyl Perfluorooctanesulfonamido ethanol (N-Ethyl FOSE) conducted at Covance
iT Madison. Dr. Andrew Seacat was appointed study director with Dr. Marv Case as
0F altemae.
La ]
Objecti3v-e To investigate the backgroundserumand liver PFOS levels in naive
S
ratsofdifferent age groups from different sources. Information on the various
=
diets supplied by the different breeders is to be obtained. Deanna Nabbefeld was ~~
appointed study
Objectiv4e - To
director with Dr. Marv Case as alternate.
investigate the possibility that PFOS exposure
is
stemming
from
tainted feed, exposure in rat roomsor a combinationofboth. Dr. Marv Case was
appointed study director.
3gaMthEenrevdirionnombejnetcatlivAensal1y-t4ical Laboratoirsy responsible for chemical analysisofsamples ~~
or
=
3+ 2=x ~~ = Es
--1
5
`The study to examine objective 3 is complete and is the focusofthis report. Once data addressing each objective are available, reportsofthe various studies will be generated.
Study Objective: "This study was designed to determine what perfluorooctanesulfonate (PFOS) levels, if any, can be detected in untreated Sprague Dawley rats of three different age groups acquired from three different breeders. In addition, information on the rodent feed provided by the various animal suppliers was obtained.
&epa-ors
nn
000811797X
--
03984
DT21: T-6316.9;FCLevelsofNDaRARi FaTtsv 5C/O14De/E9.9
Research Client:
3M Specialty Chemicals Division 3M Center Building 236 Saint Paul MN 55133-3220
Sponsor:
3M Toxicology Services
Building St. Paul,
220-2E-02, 3M Center MN 55144-1000
Study Location:
33MM SCternatteerg,icBuAilltdeirnngat2i7v0e-TSoBx-i1c8o1logy Laboratory Saint Paul, MN 55133-3220
Study Director: Alternate Study Director:
Deanna Nabbefeld, MS Advanced Toxicologist 33MM CMeendtiecralBuDielpdti.ng/T2o2x0ic-o2lEo-g0i2cal Services Saint Paul, MN 55233-3220 `Telephone No.: 651-737-1374 Facsimile No.: 651-733-1773 Dr. Marv Case Corporate Scientist 33MM CMeendtiecralBuDielpdti.ngTo2x2i0c-o2lEo-g0i2cal Services Saint Paul, MN 55233-3220 `Telephone No.: 651-733-5180 Facsimile No.: 651-733-1773
Summary:
(`T6h-e8 lwieveerksosfomlda,l1e0a-n1d4 wfeemeaklseoulndtraenadteodldSprertaigruedebDraeweldeerysr(atOsoRfBt)h/r=ee9-di1f2femroenntthasgeolgdr)oups frPoFmOSthraeneddoitfhfeerreknntborwenedfelrusor(oBcrheeemdiecraAl ,(FBCr)eemdetearbBoliatneds.BrEeaecdhebrrCe)edweerrweaes xcaonmtiacntfeeoddrto provide information on the feed provided to their rats while at their facilities.
MLiavleersraftrloimveBrsreferdoemrbAotahnsdouBrrceesedineCcrreraastesdcsoingtnaiifniceadntdleyte(cpt=ab0l.e0l5e)veilnsPoFfOPSFcOoSnce(n2t15rpaptbi)o.n PwiFtOhSaigne,li(vselrospfer=om5f1eBmraeleedBerreAedmearleA;asnldopBere=ed13e3r BCrraetes,dheCorwemavleer),.diTdhneoctoinncecntrreataiwosinteohf Cagema(lseloOpeRB=0S.h6aBdrteheedheirghAesftemPaFleO;Sslleovpeels=(-a3v.e7rBagree=ed32eC7r+fe1m1al5ep).pbT)h,esilginviefriscoafnBtrlyeheidgehrer mtehtaanbtohleitleesveclosuflodubned dientBerceteeddeirn AthemaBlreeeOdReBrSA(oarveBrraegeede=r1C45ra+t l1i2veprsp.b)N. oNoPFotOhSerorFCother FC metabolites could be detected in the livers of Breeder B rats.
Page 20019
03985
DT21: T-6316.9; FC LevelsofNaiDvReARFatTs5C1O4/D9E9 saBurrpeepefldeideerdfebAeydrCaBlosmaurppepalfieneCddyfbe(yeCdiCtAoymCsp,uapSpntlyaiteBed C(b)Cy.iCtFyoimBs,hpmaSetnaaytleAiBs)(.CiBtyreA,edSteCarteraAt)s. aBrreefeodderfeBedrats the fifth listed ingredient in feed A and not present in feetdheB.primary ingredient in feia c PfFeFrmoOamSletihrsaertseseliedtaadstoead,esiitn5a0tphp1ee0maiarllsekasasnedtr/dhoeorgurgtehreaP.nFsfOPeoSrsrsegidrbeilanetlueyxtpeblriaoontaacticounmsulfaortethiisn mdiaflferreantcse, awvhiltehaiyn Stbhyorrmoeeuagrnhodduernniutnrascrehydoewmx,uclrtmeitapiyloencltithearns.maAllesor,asf.emIatlies rpaotsstumlaaytemdotrhteahteriepasudhpilsmyeiaenlx.cfreaemntaelienPsgErteGhdaSiyenhtowien PhFeOSposissiablislotbyetihnagt c3oMnsimdaeotrneetrdai.ianl iFCusaenddtt0hcuosalteatdhtofetehde bPaFgOsSanfdoutnhdaitnhiusntisreaastoedurrcaets.of
Methods:
owTrhedieerkrtseydoSlpfdrr,ao1gm0ue-ea1Dc4ahowwfelteekhyseroafltosdll(afoniwdvienomgdablrreee,teifdrieevrdes:fbBerrmeeaedleeedpree(rrAOaRBg)e.gr=o9up-;12agmeognrtohusposl=d) w6.e8re
lborceaetdieornowfasfeaesdkebdreteodeprr,oveiedde
the nameofthe feed prov,idBerdeteodetrheBir ingredient lst and information on feed
aantds,BnraemeedaeCdr packaging. All
Bach
WNainotikmfaiolnosdornoeermhawiaontueerdo,ifontathrheriirvratslhhaaintpp3thiMan,tg prcarotonsvtiawidenerederiswnebtiehgtehwseehdei,nppgarironrsgisvlcayolenatxtaai3nMersa,ndwacsutfhuarsniizsahteido,n.
LCaObso.raSteorray a-nFdluliovreirneweArnaelhyatrivcaelstCehdemainsdtsreyntTetoaKmri(sFHAaCnTs)e,no, r3FMCmEiannnvaeildrysoiansnm.den(etuAatlhanisedby
TSdtehasetcirrsiatpittceaiolofnsicoghfnaitnfhgieceaannoacflePyosFfiOsSremseultiethvseolwdsacwsaandsebtaeenarflmoyiuznneedddibunysAippnlgpoettnhidenigsxtau1gd,eeEnxtspeTr-iTmeesntta(lp s=ec0t.i0o5n).,)
concentration (ppb) and calculating the slopeoftheline. in months versus PROS
Results:
LaboratoryData
Raw oes pnd &
data, including body frualltidoantsa ctaabnleb,ecfaonubned
wieniAgphpse,ndliivxer1. weRiegshutlst,s loifvethreboFdCy found in Appendix 2 - a Sum
awneailgyshits,raitnicolsudainngd
PFOS graphs
Hansen.
mary Report propared by lors
snLtoiutvdeyyretpslabamenpel(nseesaenawbleayrczeekdg.dreoeuDmneedpdesnemdcotiisnotgn,mopenga.tnhie1n)g,ofusuetlrcaaoPnmdeFsOothSfusaonbaajlneyacsltiyiszveemds
first. 1, 2
Sera and 4
samples have of the guerall
ay not performed.
Average PFOS liver levels standard deviation for the various age / sex/ breedergroups
Page3or19
03986
included in this study are as follows:
DT 31; T-6316.9; FC LevofeNlaDisvReARFaTts5C/O14D/E%
BBrreeedeerdAAe66--r88 wweeeekk oolldd FMaelmea.le
43.04 4.07 ppb 66.+6 19.27
Breeder Breeder
A A
10-14 week 10-14week
old old
Male Female
103.0 21.33 791.2.906
BreeAdOeRrB Male Breeder A ORB Female
145.0% 12.42 67.8429.63
Breeder B all groups
< method detection limit (15 ppb)
Breeder C BreedeCr
6-8 6-8
week week
old ok
Male Female
~~
60.2% 73.0%
12.21 12.29
ppb
Breeder C BreedeCr
1 10-10 4wweeee- kkooll1 ddFMeam4 lael~e ~759.2.7242242..4705
Breeder C ORB Male BreedeCr ORB Female
3270411537 65.6% 27.24
iTBsrhe5e1edwreahtrielAeotfahcnahdtafBnorgreefeedomefarPlCFeBOrrSaetesldirveeersrpceAcortnaictveesnlitysr.a0t.Ti6ho.ne TwsihltoehpesaolgofeptehisoefdtelhipneielcitfnoeirnfmoFailgeurBersee1 daenrdA2rfaotrs
BreedeCr rats is 133 and 3.7 respectively.
rmaleand female
Page dof 19
03987
Figure 1
i=w12o00 2 a0 3
g =c=wooo
a0 :
DT 21: T-6316.9; FC Levels of NaDiRveARFaTts5C/O1D4E9 PFOS LivBerroCoodnocrenAtrations.
3 "hae (Hooke)
ee f-osre ozs
as
#7we=e68kweesks old: 12weeks= 0-14weeksold, 45.5 wee=kORsB.
Figure 2
PFOS LivBerreCeodnecreCntrations
_ snooo]J
===I =
Siam35oe00 30
Em x
8
1
1000.
= = La
*
=a
Ri J
"
A
"Age (Weeks)
SF-i0m85a05
=-a7x+78.039
=
+ aFaeno [-- tier ans
ws
"7wee=k6s: wecks ol; 12 weeks= 10-14weeksold 45.5 wee=OkRDs.
sTihgenifiinccarnteasbeetiwneePnFaOllSthcroeneceangteragtrioounpsfo(upnd=
in the
0.05).
livers of male
The increase in
Breeder A rats
PFOS levels in
was
male
Breeder C rats was not between the 6-8
significant and 10-14
between the 10-14 week old groups.
week old group and Comparing males to
the ORB group, females at each
but age
Pages ol 19
Lu3958
DT215 T6316. FC LevoeNlaiDvsReARFaTts5C/1O4/D9E9
group the from both
Breeder Breeder
A A
10-14 week and Breeder
Colddigffreoruepdssidginfiffeirceadntlsyi.gniCfiocmapntalryianngd
the ORB Breeder A
groups rats to
aBPgrFeeOlesSdeexrlivgCerrorcuaotpnssceadtnitedraacnthoitoangssei/tgnshiaefnxicdgairndotulmypa,ldeimfafBlerreeeiBdnreerPeFdAeOrrSaCtsloeOfveRtlBhs ebrsaetatswmeheeaandgsetihggenrioftuiwpc.oantbAlrlyelehodietgrhhsee,rr
Results of the T-Tests are shown in Table 1
STaTbAlTeIS1TICS T*-=TEstSaTti-stitcwaol-ltyasiilgendifdiisctarnitbution, paired, p =0.05,
[Breeder]
"Age/Sex Group Comparison
[Ma 68% l Tioe tank 10-14wha ORE
[6owkT & [65 Fo[10m-1a4wek |oRB [10-14wk|a ORS
[| enka[65wk viaioromae[OF&
ons |a 66 wk
ors
I
Sredercompareon
[Ma [5 wil70e -14wk [ORBT[6:6 wk A [868wk|a 10-1wkorB | 68wk
FJor0m-1aawek JORBE |s 10-1awore
|woovooomoooomomo|n
vs.
`BreederFeed/Diet History
pBarcekeagdeeAdrinraptaspaerre bfaegdsfeceodatAedsuwpipltiahedpabpyecrocmopaatinnygAmat(ceirtiyalA,prsotadtuecAe)d.byFe3eMd.A Tihse feed ingredients, as listedbycompany A, are as follows:
BgrHoAu,ndfiysehlmloewalc,oranl,falwfhaemaetalm,idcdalniengmso,lassosyebse,ancamlecailu,macnairmbaolnaftate,prsaelste,rved with peaynatnootchoebnaaltae,mifnoli(csoaucricd,eorfibvoifltaavmiin,ncBh-o1l2e)c,albciiofteirno,lD(Lsoumrectehioofnivniet,amcianlcDi-u3m), vitamin oAxiadcee,tastoe,diduim-asleplheanittoec,onpihceortyinlicacaectiadt,e p(ysroiudrocxeinoef hvyidtraomcihnlEo)r,idteh,iammeinna,dmioangen.esium
Pages or19
03989
DT215 T-6316.9; FC LevofeNalDivsReARFaTts5C/1O4/D9E9 sdiuioldmfaeattteeh,,ylmzpairnnicgmaoixdniiodnueo.sl obxisiudlef,itceo(pspoeurrcseulofaftvei,tcaombianltKcaacrtbiovnitayt)e,,ifcerornousdicoaxribdoen,actae.lcsiiunme
HlpBoaerceokeadategreeBddinwaitptsahpaaerre3bfMaegdsma.fteeeIrdtiaiBsl.usunTpkhpnleoiewfdenebdaytBcthoiinmsgptraiemndeiy,eBnho(wietvyerB,, wsthaettehBe)r. orFeneodt Btheissebaasgos
follows:
t,a sed by company B, are ac
bsgroreyowbueenradsncdoorilmi,e,cdsoyoemyabsgetlaunitoemdneimazelea,dlsg,arlcota,ulnLc-diluyomsaitcnsea,,rwbDhoLne-aamttee,tmhiddidclailncgisa,maplfhaolsfpahmaetael,,
mc1eaanclatudiimvoanptaeendstaoontdihimeanlatset,erroilbo(fsloauvirnc,eotfhivaimtianmmionnDosn),iavtiiteoa,nmiipnnyer,EidvsoiuxtpiapnmleienhmyeAdn-rstoc,cehntliaaotcreii,sd,e,
cvoitpapmeirnsBuulfzasteu,ppzliuenmmcebonixtsi,udlec,iaelcccailoucmmipuclmaerixbood(nasatoetu,er,cceombaoafnltgvaicntaaormbuiosnnaoKtx)ei,,def,olfiecrracoiuds,sbuiloftiitne,,
spBaorclekeawgdheeedtCrhienrraato"facr1oe0mtfmeeadr3cfMieeadmlayCteasrcuicpaeplpltiiaebudlsebey3dcipnloytmhlpeasmaeinnbaCaytges(d.cpiyapCe,rsbtaagt"e,C)I,tsFeuendkCnioswn at this
as listed by company C., are as follows:
The feed ingredients in ocd C.
s&foirsyohbuemnaedna#lo2,il,yseoslayllbto,ewadniccomarela,clig,urmaolpfuahnlodfsapwmhheaoatllee,, cogaortrosn,ungwldhueltaiemntemmseitadoldn,lei,gnrvgisot,unBdrewwheorl'es wdhreiaedt,yeast,
premixes.
amin and mineral
Conclusions/Future Directions: SA`2uhaepnldpivlBebrrseyoeBfrdBeeCreredeeidnrecrrAeBahsarevaitensdPaepFtpeOecatSarbfclroeencloeenvfetlrFsaCot.ifoPnLwiRviOetrShsabgfuero.m LmiavleersatfsrosmufpepmlaildebryatBsreeder Soiirngecneridefeairscsain.ntglFyf,ermioanmlP6eF-r8OaStwseseukpsploiledd tboy1B0r-1e4ewdeeCerksacotludalalnyddedcertcertaehsaeessiesnllgiegsvhletilygs,htpalhylattiehnaon1u,gdhslrnioghtotly ePpFrOeSe.greOantelyhbyipooathcecsuimlsuilviaesrttechsoanticnPemnFtaOrSlatirisaotnrse,wliwethahsieladegeiin.n tfhFeemraomlimelktrahatnessdeliotdrdatotaer,sa1nssoptpeorae3rJscaaptrhough rftahetsem.aplueprsaisn afreemamloesrthraetadhialvyeabboeretoacnldeanrurPsOedSmutlhtrioplueghluirteirnsa.ryAneoxctrheetriopnosthsaidnbilcnyssfseortuho.atto
tn`hAeolutughnohkutgnthohwattnhe,issoohnuemrechaeyslpoomtfahePysFibOseSiasnltehiaandtgirnPegdOi1e0SntthicseoclnoitnvaetiranicionnnegcdPeinntrtahteiroantscihdoenwti(fOiebdjeicnttihves 2s)t.udIty is OS. While not present i se
PageT0r19
03990
DT 21; T6316;FCLevoefNlaiDvsReARFaTts5C1O4D5E9 Breeder B rat chow, fish meal is the primary
the 5* ingredient listed for Breeder A chow.
ingredient Analys
listefd orBreedeCr
rat
chow
and
musaeydbbeyeoabtcahinaendimaanld barnaeleydzeeidrscuforrreFnCtlcoynutnednet.rway. isIon fatddhietiFonC, csoanmtpelnetsofoftthheerfaitsch hmoeawl
irsAenspouotltthksenrspoecowtsnisoani,be3hsMiosustruicpmpeeloiiffesp3PMaFpmOeaStrecirosiatathliepinasgaptseoorCcuoosametpdiainnngybuaAsg,esdtfhne emaendufbaacgst.ureArsofsFtaeteeddinA.theIt information on feed packaging will be gathered ina sparateosrtFudeye.ds an Further
Signatures: Prepared By:
`DAedavnannaceNdabTobxeifceolldo,gMisSt
Date
Reviewed By:
CDor.rpMoarratveCSacsieentist
Date
Dr. Andrew Seacat Senior ResearchToxicologist
Date
Pages or19
03991
DT 21: T6316;FCLevelsofNaiveRats CODE
Appendi1x
DRAFT5/14/99
PT
[Body [tiver
FFGS LiverConc. (ng/g o pb)
wi(9) wt (g)
aL ] [T ewe | 77I 7 830
individual _|Ave/Grp [Ste/Grp
T oa T 5 T CT
~~ as
leaawwiss || 116719.69 783005] ooooue loows | 1516 ceo ou
5
las A C loaves | 1E 066) 806] ooss 47 IF loowks | 1683 713 os
65230) 3
4h9e JF|F lloooommss || t14e17s]s] a7o4tg oooo #10 Floss | 1507 a7 oos
9659.44 512)
a[A1T2T [MWr [1[00--1144wwhios|] 32268735] 1025656 0o.a0d0 ata IM [10-14vhs| 3567] 1310] oa
71040009 96.8
JE
aat1s6IMm_J[1100--1144wweiss) 3a6s5.s6 1133625]5] ooomd
5a07.]1
4[31776 JF[F [1f01-01144wwhhsa|] 129206..91] at [F [10-14who 2034]
760.3425 513
00.000 00a
752339] 7]
oq
4A2109 F[_F [[1100--1144wwhiss| 2190549]6 771048] o00o0oq 6o04
4[p2e2t I|MW a2 [M
[[oorRsB fore
| 3[442096]2 | 051
1007080 1089
0o020ad9 0027
T133g0 1519
3
3
2lh2254|IM lfoornss || s46a0r2]] 13`.5992]4 oooooo 11640900)
2p275 las
[[lFoorrss IF lors
| [s3i4z2 | ses]
1162078d] 1450
o00oa oat
T6o.o1] 678% 450
a209|[colrores || sasasda]f "ORB = od roired broedors
11134406
oooomo
5553.0)
[MeOtuhtoderde-tencottiuonsemdiitn c(aMlDcLu)a:tiPoFnsO.S = 15ng/gor pps
Pagevor1s
03992
DT21: T-6316.9; FCLevels ofNaiDvReARFaTts5/C1O4D5E9
Appendi1x
PeeEa wt (9) | wt (9)
iPnFdiOvSidLuiavlerGo_n|eA.ve/(Gnrgp/g[Sotra/pGprbp)
[LBT I1[Fowie1| ToI ad] 51 45 T 004g T <L] T
B2
6-8wks | 201 9.34 0.046]
<MOL
88s3 os
6s-a8wwkss | | 220030.94]] 88.3580]] 00..004412 saws | 2042] 017] oo
<<MMODLL <moL|
6[B7s ||FF [[osoowwkss [[22104380 77.1692 0030% MMOB!L <b
Bs [F loswks [2128 71a] oc
<M]
BBO10 |[rF l|e6s-8wwkkss || 220000:0 78.0380]] 00..080!9
<<MMDD!L
BB1112|W [1010--1144wwikss|| 33000763]] 712035.69] 00.0643d9 N<MDDI
18143 [[MM [[1100--1144wwhhss|| 32000480]] 11102065] 00..00037] v<wooll]|
15| _[10-14whs| 306.0] 11.38] 0.00 B16 [F[101awks| 2377] 6.68 0028]
MD! <MDL|--<MDI
BB1187 J[FF |110o--1taawwkss|| 22340171 772.01]4 00.003000 MvDLo]
B82109||F _||1100--11aawwhkss|| 22530643 7s3s3a) 00003ag <wMoDI]
BB2212 [[MMm [[oORRBB || a47e027s]| 1174.0900] 00.000328] <MwDoLL]| <Mol|
B6223[vM J[oorns || aassazlf 11455552 oo0a0a]s <wvooL]|
Bos m Jors | a7eg| 11.19] [B26 [F [ore | a7] 1045
ocea 003]
vou] MDL] <WDI
Bee2s7 [[F Fo[rorss | 2[825672] 1082574] 0o00o0 M<DwoI|
BB23o0 [JFF Jorosr || 23086592] 11o.o00t] oooafd vvoou
- ORB = old retired breeders
[Method detection limit (MDL): PFOS = 15ng/gorppb
Page 100619
03993
AppendiDTx121; T-6316.9;FCLevels ofNaiDvReARFaTt5sC/1O4D/9E9
IBwe(t6)TIwot r(9)
i[nPdFiOviSdLuiavler Co_n|Acv.e/(Gnrgp/g[Sotrd/pGprb)p}
[L ecTz [lcoa8vwkkss|| 2200240f2T] T109:614T8]] T T 00.0045g0 T 6w3g0 T7
lca lca
je6-a8vwwkkss
| |
11687.45].1
1092268)
00..005580]
449727]]
cs [C6
[F
l68wks | 1836] [ews [2012]
034 973
00s 004
7. 7.0] 9 9
c7 c8
[F [F
|oawks [e8wks
|
1809 [1725
sao o40f
oo oo
89. 788]
co |r c10 JF
J[eea8wwkkss | [192050] 7 11002584
o0.o049]
5763.26]
[ert icr2
1100--1144wwkiss|]
4543 334.6]
1137..0977]]
00..003499)
545;]
z7
c13 [C14
1100--1144wwhkss|| 4405705.9] 1174.1675] o0.c039
1894.))
cts [C16
[10-14wks| 317.8) [F [10awks] 2144]
12.75) 933
0.040 oo
86.2] 529 75
3
[CC1187 [[F F[1101-144wwkkss| 22216611
889447
0.04 0.009
715] 726
(19 [020
IF |F
[10-14 wks| 10-14 wks|
241.0f 2465
11.21) 8.53
004; 0.05]
64.4 179)
lcc2a2t O[ORRSE || se6a1.r2s| 2128.2224] 00.00339 454110. 8270] 1153
ic23 C24
ors [Mors
| 4seo| | s57|
14.96) 17.91]
0.033 oad
379. 3900)
[cas ors | sae] 1824) 0.001 [C26 [F [ore [2815 10.56 0.069
2760 55.
q zz
(027 IF (C28 |[F
fore Jors
| 2s14f [2475
1881 1565
00s; 0.06
105.0 71.3]
20 |F cso |r
ors Jors
| |
328807s]]
1174.9585
o0.o52]
629300)
[*MOeRthBod=doeitdecrteitoinreldimbirte(eMdDeLr)s: PFOS = 15ng/gorppb
Page Hot19
02994
Appendix 2
ws
3M Environmental Laboratory: FluorineAnalytical ChemistryTeam
Contact: Kris Hansen Fluorine Analytical Chemistry Team Building 2-3E-09 8-6018
Studyof PFOS levels in Naive Rats
Summary report
Experimental Summary
livers
In order of ninety
to assess "endogenous" levels of PFOS in test animals, the rats from three different suppliers were quantitatively
analyzed for PROS. ofanimals received
Threedistinctages from each supplier:
of rats were represented in 6-8 weeks old, 10-14 weeks
the old,
group and
retired breeders (> Department at 3M,
14 weeks). The were sacrificed
test animals, upon receipt;
received by the Toxicology tissue samples were
delivered to the 3M Environmental Lab for extraction and analysis by FACT.
: Supplier | Location
`malRea:tfieomal
re ge[Te A iEct B il tatl C
f*e3eMd Ai.s/wCausrraenstulpyp,liietrisofnpoatpkenrocwonaitfin3gMma`tseurpiplailestomactoemrpiaalnytoAc,osmupppalniyerBoofrC, suppliersof feeds B and C respectively.
Analytical Summary
reagentL.ivTehresaemxtprlaecstswewreerehoamnaolgyezneidzeqduaanntidtaetxitvrealcyteudsiunsginhgigahn-iporne-spsaurrieng EliSqMuiSdMcShr)omaantdogervaaplhuya-teeldecvterrsousspraany etxatnrdacetmedmacusrsves.peAcntarloymteitcrayl (deHtPaLilCs-are falvuaoirlaobclheemiincatlhecfounltlarmeiponratn.tsThanedprmeesteanbcoleiotresabwsaesncaesocferottahienrekdnboywinnspection.
03995
DT 21: 6316.9; FC LevoefNlaDvsReARFaTts5C/1O4D9E9
Appendix 2
Results Summary
smiaglneifriaRctaasnttflreionvmedrosBgrfeerneoodmuesrBlrCeevceeoldnsteoAarfiPnaeFndOdSsBi.grneLieifdviecearrnsCtlcyotlemlsoetcratenediPmfFarlOosSm sothlhdoaewnsetadgnryooutphoefr
group in the study.
the
The limitof
ldievteresctoifotnes(t1a5npipmba)l.s
from
Breeder
B
did
not
contain
PFOS
above
trhoeugyholuyPnFgcoeOrsSrtellmeaavtleeledsrwiainttsth,h6et-hl8eivwaeegreesokofsfmtoalhdle,eacnroianmttasalifsnr.eodmTthBharetelesed,aeslrtivAPeFraOsnScd,olBlwrehecietlededetrfhrCeom
old,
ret`iTrheedlbivreeresdoefrthmaelfeermaatlse
contained rats from
tBherheigehAedsteancrodncBerneterdateiroCn.were
determined with age.
to
contain
very
consistent
levels
of
PFOS,
showing
no
correlation
No identified
oitnhtehreklniovewrnsafmlupolreoschaenmailcyazledcoinnttahmiisnsatnutdsy.or
metabolites
were
Graphical results and a full tableof results are attached.
ofPFOSCurarnednteltyh,ymle-tFhOoSdEsaalcroehboeliinngsdaemvpelleospoefdcfhorowthefraonmalcyascish osufplpolwielre.vels
ESxapmeprliemepnrteaplaration-aqueous extraction
samp
les,
HPLC
ES
M
S:
I
on-
pai
rin
g
(etnevtirraobAnunmtyaellnytt.aemismoexnTtihruaecmtedchaftyridoornmoigcaesnarmepsaulgleefnamttaetris(exTlweBciAtt)ihvealinynionta-apragpeiHtr-sicnognrtaernoaitgoleneindct
ftlruaonrsofcehrermeidcailnst.o
a
Once the non-polar
oarngiaonni-cTBsAolvpeanitr
is formed, the (ethyl acetate),
analyte is dried, and
reconstituted in methanol for MS analysis
HPLCIEnSMHPSLaC,ndaHnPLalCiqEuSotMSofMSex:tFraocrtdiestaiinjleecdtedquaanlidtaptaisvseedwotrhkrough a raneavleyrtsee-fpohrastehelisqtuaitdiocnharroympahtaosgeraipnhitchecocloulmun.mn rBealsaetidveontotthheealfifqiuniidtyomofbtihlee etpixhmaaemssep,lbee,tthiewneaaennsatlaaytnsedtaairnsddraseortlduatiPinoFendOPSfForsOoSlaumtciahoaynraeacltnuedtreitsahttiec10aa.n5malomyuitnneutteexostf.ratciRtmeeetd.enftrFiooomnr groundwater in this analysis were matched to within 1% on the HPLC
system.
Page 130019
03996
AppendixDT221; T6316; FC LevofeNaliDvsReARFaTts5C/1O4D9E9
aannailoynziiznFagotilaolnowwitiednceghnriaHqnPugeLeuCosfesdocrppgararinamtiaicroincl,yomfEporoSutMnhSedsd,eptreoicnvtciildouendsionfag mfrolalupeoicrduolcahanerdmiicoaascl,csu.irsEagtleeenctmreoasnpsrafyo,r
Qperated detected.
at
relatively
mild
temperatures.
Molecules are ionized, possibly fragmeenrtaelal)y, and
HidoewnetvifeiErc,SatMiionSns.MteSadAoasfdsdsiinmpalEnySaMmdSod,niititaoonraiclnhagrdtaichmteeenprsirsiitominacryopfrioicnme,ratrianyiEntiSyonMtofscosmepotunsd
dsisemtabelcoltmeerbda.sredceodndwairtyhiohniigchf-reangemregnytgsasu.niqAuse atorteshueltporifmhairgyh.ionenaerregScyMrceoSlaltiteshdieounjnsai,n
achmaruac(tFceoorirrsrteeixscapmopnprdliiemn,agrfyotroioPSnF.O,O)TS,hi(9sC9,iFo,an,miSsu0,f)r(acagnoarmlreyensstipesod,ndiiinontngo4o9tto9heFirsSfOso.en)lsecstuecdhaasst5he
dB(icafocfrherreesnoptfioanttdehiensgPeFtOosSeCcFof,nSrd0oa,mr)y,otf1hr8ea0rgmacenmotmusp (iCsF.dSeOt,e)c,tedandan2d30caanmub,(eC/1aF3s,0eSd0a,m)bu.r
scehacroancdtaerriysitoinc f4r9a9gmaemnutaptiroinmaprayttfeornnsb.utoudinfdfserentthacthemmiigcahlt choampvoesitthieo.nssaamne
HPLCCsoylstuemmn::Hewlett-PackarKdeySsetrioense1B1et0a0siLliqCu1i8d Cchorloummantograph
Flow rate:
320X01m0i0 mnim, 5pm partiscilze:
SSoollvveenntt BA::
M2e.0thmanMolammonium acetate
Solvent Gradient: Hold at4900%%Bt0 9fo0r%3Bmiinnut8e.5s minutes
HRoeltduamt41004%0%B fBorin| |mimniuntuete
Injection volume: Run time:
10pL. 13.5 minutes
MElieccrtormoaspsrsaQyuTaatntrdoemITMAaPsIsmSapsesctsrpoemcettreormeter mass spectrometer Mass Lynx 3.1 software
Cone voltage: Collision gas energy:
30-60v 40.V.
Mode: Source
block
temperatuerlee:ct1r1o5sprCay
negative
PDersiomlavraytIioonn: temperatur4e9:9 250C
Page 14of 19
003997
AppendiDxT221; T-6316.9; FC LevofeNaliDvsReARFatTs5C/1O4D9E9
Daughter Tons: Electrode:
80,99, 130, 180 Zspray
QualitAyllcaonnatlryoslesswuemrmeacronyducted with a moderate to control. Duplicate matrix spike analyses were condu
high
levelofquality
`ewaicthhignrtohuepoacfcaenpitmaabllse.raEnxgceeopft8a0s-1n2o0te%d, in the resultcstteadblfeo,rroencoevaenriiomsalwpfoeom
manoanliytsoiArs iconafsltitwbroruamcteuinortnvaeclshdebrcriakftc.skteQatuniadnnagtreidaawcthaiosgnrawonuaapsloybfzaessdaemedpvloenryli5n-e1a0rsraemgprlesessitoon
PFOS was based on the responseof 3-4 daughter ionseosf.thQeuaPnrtiimtaartyioinono.f
Results
See spreadsheet attached to this report.
FACT Km.emHbaenrssenparticipating:
L. Clemen
HM.. JEollhenfssoonn
G.
R.
WLyanngneenburg
LA. Smith
S. Heimdahl
Page 150r19
03998
Bete t
B=
OT Cle nT pcrrs
ame
Appendi2x
Bee...
HEE
Eetsmim
=
PerfEaLE ETaEaE ErRE SLeeEei Sr
CEELlEe (E EE EE EE EE
LEE CELE
TLTETEIEEE E EEE
g E iEe JRe EITEEE
$ SRE mm
Page 160019
|
rac im
Male Naive Rats-T6316.9, FACT070998.1
Er----
g
m 2Ii mvr TR ms 7 | | g==E
]
Hemr l 8 o 2500 =m
e es t BH |WBreeder 8|
Poml b r= moes
a TT Wo|orat
oa] Hl
Tea
cusses,
`Age Group
gg T EE rs T Tues 2S, `rAoveoraargePGFrOoSluepvels esta wer a <P73 OL 4p) determined frompopulation n = (per AgeGroup,perSource),except Breeder A6-8Woek MaleRats
cos
Female Naive Rats-T631 6.9, FACT070998.1
$ . -TrrEE =
2 wm
d Sealie LRs a
booat
g=a
E20
8 5
8
2 mw
Seger
: ot
atsSE
io
HRA
TSHiRnEe
Bis
[Bbresder | =
ez
RE
EL
[MBreederC|
2:3 E e E EPr E eEE e nr EEE r Bom y gs
`Week 6-8
Ear
`Week 10-14 Age Group
A
Old Retired
TT =r]
MAeppenedinez
DT21 T6F1C e6ws9a,vDeRWa TCosD:
Naive Rat Study-T6316.9, FACT070998.1
wm
HH E[=] SATRr E
So
3
=
| iijEio .Sr SPoTuRe=ES HSoEaEAa) S33L3lAF] E[FT eema] E8=5 2
Pl 100.
|
E
= "I!l Thii[| EE
]
sex
Bear
TE qT
oyBy te Awere odFed A CoAgmepaGrouplSooufrfactes
ote
Seve
2SBSBeeroeecnrerrCrrtotoy(CCtp Ryy08,rSoite sttatOBwwwooa1eremsopedFpFPednot, ApGroupSrc, net rs A 64 Wok
3
s
Fue10otis
.
DT2L: T-6316.9; FC Level in Naiv RatsTM Further InvestigationofRat Chow CODE,
DRAFT 514/99
,
3M MEDICAL DEPARTMENT, TOXICOLOGY SERVICES
E
fecal (FC) RLeopwort for StfiveudyRatsNo. =T-F6316.9; IDnTve2st1igation of Rat Ci
-_--
Background:
bInuradnenatftoeumnpdt tion cdoentierromlirnaetsthienvsooluvrecdeoinfsloomw-ele3vMelcpoenrtfrlaucotrdoioecttaarnyessutuldfioensa,tea (PFOS) body
comprehOebnjseicvetipvl1ean-wTiothitnhveesftoilglatoewipnogteonbtjieacltisvoeusrcaensdorfescpoonntsaibmiilniattieisonwawistdheisnitghneeds:tudy
ChoouvsainncgeaMraeadiosfotnh.e Dcru.rrAenntdrdieewtaSreyasctautdiweassoanpppeorifnltueodrisntautdeyd dtiersetcctoormpwoituhndDsr.at
cMoaorrvdiCnaatsiengasaanldtecmoantdcu.ctJiinmg Wdoirletcetrsa,ir3moMniEtnovriirnognmaenndtawli,peisraemspploenss.ible for
OPbFjOeSctseiev2nei-nTcoondtreotlerrmaitsneiniftchoenttwaom-iyneaatriodnioeftafreyesdtuidsyleoafdNin-gEtthoytlhe low levels of
PMeardfilsuoonr.ooDcrt.anAcnsudlrfeownaSmeiadcoatetwhaasnoalp(pNo-iEnttheydlsFtuOdSyEd)irceocntodrucwtietdh Datr.CoMvaarnvceCase as
alternate.
Orabtsjoefcdtifif3ve-reenTtoaginevegsrtoiugpatseftrhoembdaicfkfegrreonutnsdousrecresu.m IannfdorlmivaetriPonFOonStlheevevlasriinounsaive.
d`iaeptpsoisnutpepdlsitedubdyydtihreecdtiofrfewrietnht Dvre.ndMoarrsiv Ctaosbeeaosbatlatienrenda.te.Deanna Nabbefeld was
Otabijnetecdtfieve4ed,-eTxopoisnuvreestiingartaet trhoeopmosssoirbialictoymtbhiantaPtFioOnSofebxoptohs.ureDri.s sMtaermvmiCnagsefrwoams
appointed study director.
3gaMthEenrvedirionnombejnetcatlivAensal1y-t4.ical Laboratory is responsible for chemical analysisof samples
`cTohnetafmoicnuastoifotn.hisThreeposrttuidyttoo ceoxvaemrinteheodbajetcattioveda3tiesecxoammpilneitnegatnhde ipsoscsoivbeilrietdy oinfarsaetpcahraotwe.
report. studies
Oncefurtherdata wil be generated.
addressing
each
objective
are
available,
reportsofthe
various
Tcuhrere3ntMlyEinnvtihreonpmreoncteaslsoLfaabnoarlaytzoirnyg--raFtlucohroinwesAanmapllyteiscafolrCPheFmOiSstarnydTietsammet(aFboAlCiTte)s.i
AAtutgaucshtedofis1a99p8re(lAipmpineanrdyixsu1m).maSraymprlepeosrotfgreantercahtoewd bfyro3mMCoEvnavnircoenmLaebnotraaltoLraibeosr,aHtaorrlyainn
aLlacboohroalto(rEie(sFOanSdE-NOIHH)bweyrGeCaMnaSl.yzeTdrafocres2o-f(NE-ettFhyOlSpEer-fOlHuowroeorcetafnoeusnudlfinonaasmaimdop)l-eetohfyl
Covance airborne
chow. particle
This and
prompted the 3M Environmental wipe samples on site. Preliminary
Lab Field Group to conclusions drawn
collected by the
air,
Environmental `maybe due to
Lab based onthedata respirationofairborme
to date are that "Contaminationin EtFOSE-OH, from transfer of test
control animals material from onc.
afnoiomdaslotuorcaen"o.ther during sample handling and possibly from sporadic contaminationofthe
Page ofS
4003
DT21: T6316. FC Levels inNaveRats =Further InvestigationofRaDtRCAhFoTw5C/O1D4E9.5 Appendix 1
Lisa Dick/ Kris Hansen Fluorine Analytical Chemistry Team Building 2-3E-09 612-778-754/0 612-778-6018 Iadick@mmm.com / kjhansen@mmm.com
Preliminary summary report: Further Investigation of Rat Chow Summary: In June 1998, several rat chow samples from Covance Laboratories and 3 from NIH were supplied to the Environmental Lab for characterization of fluorochemicals that are being monitored inanimal studies conducted by 3M Toxicology. A single sideofthe bag chow sample from Covance was determined to be of significantly higher contamination than the samples from NTH. Because contaminants were found, air and airborne particle and wipe samples were collected on site by the 3M Environmental Lab Field Group. So that results could be based on more than a single sample, additional bags of chow from Covance and Vendor B were also analyzed and are reported in this summary. In the chow samples, 2-(N-ethylperfluorooctanesulfonamido)-cthyl alcohol (EtFOSE-OH) was quantitated by GCMS. EtFOSE-OH was not present above the detection limit of 1 ng/g chow in the triplicate analyzed samples from three Covance bags (labeled meals for June 9, June 15, and June 21, respectively) or from the single Vendor B bag. Triplicate chow samples have not been analyzed for PFOS. Air and airborne particle samples were collected at Covance by Jim Wolter and Kurt Oldenburg of the 3M Environmental Lab Field Group. Air was passed through charcoal and glass fiber filers. Detailsofthe sampling procedure can be found in a report by Jim Wolter and Kurt Oldenburg. The largest volume air samples from the backs of rooms were collected on glass fiber filers and analyzed while samples from the front of each room were collected on charcoal and analyzed. Wipe samples from all rat backs were analyzed but wipes from cages, doorhandes, and floors were not. In air samples taken from Room 3045, EXFOSE-OH was detectedby HPLC/ESMS at levels below the practical quantitation limit (approximately 0.17 ppb/L) but above the method detection limit (approximately 002 ppbL) in some samples. Perfluorooctanesulfonate (PFOS) was not detected at levels above the blanks in any room. Analysis of wipes from the backs of rats in Room 3045 contained a measurable amount of E(FOSE-OH, whereas wipes from rats in other rooms did not. Due to variations in sampling technique, wipe samples were not quantifiable. Wipes from animal backs were not analyzed for PFOS.
Page20f5
04004
DT21: T-6316.9; FC LeveAlpinspeNnardviexRa1ts Further InvesigationofRaDtRCAhFoTw 5C/O1D4E5.9
Chow source, Vendor B Covance ~ June 9 Covance -- June
15 Covance - June _-- s21r
E(FOSE-OH Levels in Rat Chow
Number of samples
Matrix Spike Pass
EFOSEoH
3 3
sample sample
Non-detect Non-detect
3
sample Non-detect
3
sample Non-detect
Covance
Sample
EtFOSE-OH Location
LevelsinCovance `Number of
Lab
Environment EtFOSE-OH
detection
Room 3045Air Samples
samples Zoutof
0.02 ppblL <detect < 0.17
(GF and charcoal)
Room 3004, 349 Air
16
PPUL Non-detect
SaRmpoloems (30G4F5aRnadtchBaarcckosal)
6
Detect (8 ppb/wipe average)
--m Room 3R00N4,034R9uRBataBcackkss 1122
NoNonn-ddeettecett
SEaxmppelriempernetpaalrastuiomnm:aMreyt:hylene chloride extraction
and thenAncaolvyetreesdwweirteh 2ex0trmaLcteetdhefrr.omNocnh-opwolbayr oardgdiatniiocnanoafleyttheesr.traSnsafmeprlfersowmetrhee cwehiogwhetdo
the organic Fifteen mL
olfayeetrh.erSwaamsplreesmowveerde
asnhdakbelnowfnord1owhnoutro
and 1 to
t2hemnLc.entrifuged
for
30
minutes,
extractiGolnaswsithfilmleerthfainbeorl
aonrd etchhearr.coaAlll adosfortbhaetecsharwceorael
split and inside the
then prepared by sterile tubes was
extracted. Final sample volumes were 1 mL.
GC: Characteristic retention times
chromatIongrgaapshicchrcoomlautmong.rapInhdyi,viadnualalcioqmuoptonoefntssamopflethies sianmjpeclteedadasnodrbvatopotrhiezesdtatoinotnoaray
phase of the column based
column. As the on physical and
temperature is raised, chemical characteristics.
components are eluted from the An inert gaseous mobile phase
carries and gas
tphreescsoumrpeoanreenatdsjutshtreodutgoh
otphteimciozleumcnh.ro~maCtaorgrrearphgiacs
flow rate, separation.
column
temperature.
MS: Electron Impact feloercmtartoinsoO.nneoIfnmoeatdthdheiortdiforfnaotrgompetrnhoteduficooinrsnmgatthiiaootnnsmoafoyfar bmmeaosllseacrsgupeleracrtorriaosnim,saalblyseerbriotemhsobanafrtrdheemacemtnoitloenwcsiutllhaearedniseortngoeattnhidec are useful for compound identification and quantitation.
Pagers
G04005
DT21: T-6316.9; FC Levels in NaiveRats~Further InvestigationofRaDtRCAhFoTw5C/O1D4E9.9 Appendix 1
HPLC: Characteristic retention times
chromatIongrHaPphLiCc,coalnumanl.iquBotasoefd eoxntrtahcet aifsfiniintjyecotfedthaenadnaplaystseefdortthhreousgthatiaonlairqyuipdhapsheasien
the column amount of
relative to time. For
the liquid mobile phase, the analytei example, in a standard solution PFOS
retained for a may elute at
characteristic 10.5 minutes,
RfiebteernstiinonthitsimaneaslybseitswweeerneamasttacnhdeadrdtoPwFiOthSinso1l%utoionntahendHPthLeCansaylsytteem.extracted from filer
ES/MS: Detecting and monitoring molecular ions
analyzinFgolalowwiidnegrHanPgLeCofsoerpagraantiiconc,omEpSo/uMndSs,prionvcilduedsinga frlaupoirdocahnemdicaaclcsu.ratEelemcteraonssprafyo,r
otenmepeorfattuhreess.ofteMsotlieocnuilzeastioanreteicohnniizeqdu,esfravaagimleanbtlee,d,isagnednedreatlelcyteodp.eraItneidtiaaltlyr,eltahteivmelayssmiltod
charge samples.
range Tons
m/z = 100 to characteristic
1210 is of known
monitored following fluorochemicals were
direct flow injection of observed. These resuls
the are
used
to Asnealleyctsiisonosfthoartgcanaincbeflmuoorniinteorsetdansdealrecdticvoelmypfoournqduanitnidtiactaitvees
results, that the
primary
jon
characteristic compound
of E{FOSE-OH complexed lo
is
m/z = 630 amu, acetate anion
corresponding from the
to the mass running
of the buffer:
tCh4eFc1o7nScOe;nNtr(aCt;iHosno)f(tChHi,sCHio;nORin)/thCeH;saCmOpOl'e.s. Single ion monitoring was used to determine.
Quality control summary:
sample.
Methanol blanks were analyzed Charcoal and glass fiber filer
periodically to ensure complete isolation of blanks collected on site were also analyzed
the and
found linear
to be blank. Quantitation of HPLC-ESMS data regression of 5 point standard curves from 10
for fluorochemicals is ppb to 1000 ppb or
based on the matrix spike
recovery comparisons.
thesameQsuaanmtpilteatmiaotnroifxGaCnd-oMnSthpeealkinseiasrrbeagsreedssoinontohefr5epcooivnetrsyotafnkdnarodwcnursvpeisk.e amounts in
Instrumental specifics:
GC/Mass Spectrometers
HewletCto-lPaucmkna:rd ATD 400 GaJs&ChWroDmBa-t6o2gr4a3ph0mand Mass Spectrometer
Temperature ramp: 50C to 250C @ 20C/min
Carrier gas:
250C hold 5 min Helium
Spike volume: 25uL
Oven temperature: 180C
Traplow temp: 30C
Desorb time:
10 min.
Trap fast:
Yes
Trap high temp.: 250C
Page dfs
C04606
:
DT21: T-63169; FCLevels inNaiveRas ~ FurtherInvestigationofRatChowCODE. DRAFT 5/14/99
.
Line temperature: ~~ 225C Appendix 1
Pressure: Valve temperature:
18.4 psi 225C
Trap hold: Desorb flow:
S min. 25 mUmin
Ions monitored: Source temperature:
540, 448 amu 250C
EQuMadvotltesm:perature:
125C 2598 V
Interface temperature: 250C
HPLC system
Hewleu-Packard Column:
Series
1100
Liquid Chromatograph sKiezyestone Betasil C18 column,
2
X
100
mm,
5
pm
particie
FSollovwenrtatAe::
23.000mpVMmianmmonium acetate
Solvent B: Solvent Gradient:
Methanol 40% t0 90% B in 8.5 minutes
ReHotludmat190400%%B Bfoirn3|mmiinnuuttees
Injection volume:
Hold at 40% B for 1 minute 10 uL
Run time:
13.5 minutes
EMlieccrtormoasspsraPylMatafsosrmSpIIecattrmoomseptheerric Mass Lynx 2.1 software
pressure
ionization
(API)
mass
spectrometer
Cone voltage: Mode:
-60v electrospray negative
Source temperature: 90C
TAonnasl:yzer pressure: 693.0,2x51206,m49B9ar
Electrode:
cross-flow
Conclusions: hEaWnFdlOiSnEgL,-oOaHwnd,lpefvoerslsosimbolftyrPafnFrsoOfmeSrsfpooofurnatddesiitcn cmcaootnnettrraiomlailannaftirimooamnlosofntmehayeanbfieomodadluestootoarnesstphierratidounroifngaisrabmoprlnee.
urce,
Page Sots
04007