Document 6RNg2V0rb4nqrydmYxnQVvXR

M -13^ 3MStrategic Toxicology Laboratory Study Title: Acute InhalaFtilounorLidiem(iPt OToSxFi)ciTty-7S0t9u8d.3y of Perfluoroctanesulfonvl TLIInnea--sbLLtoiiNffreeautIoCmnroyibtmeiParptri(looesjatneimcoDtnpaIlDdteee)a::nteti:ficati55oT//n11-:7/420/02900810S.31T6(P4erfluoroctanesulfonyl Fluoride (POSF) Research Client: 33SMMainSCt pePenactueilar,lBtMyuNMilda5it5ne1gr3ia23l3-s63D22iv0ision Testing Laboratory: 33SMMainSCt tePrnaattueelgr,iBMcuTNilod5xi5inc1go3l23o7-g30y2-S2L0Bab-lo8r1atory v Study Director: AT33SMMaonixdniCMrtceeoPwenladotuiMegclr,ia.sBMltSDuSeNiaepldcpe5aiac5ntri,1agtml3Pi23hse2t-.nD30t2-/.2T2Eo0-x0ic2ology Services . Study Toxicologist: TP33SMMooaismntMCtmDePeionaedctuietYclor,avrBMlaelDutNtRieeldpe5Tias5nuer1gtarm3nr2c3ee2hr-n,30tF2P-/2Te2hElo0.lDo-x0w.ic2ology Services 001525 T-7098.3 Acute POSF Inhalation Limit Test. Final Report Summary: ocaacpwsrpdweueoninonxepargeavinalyrlycmnrsnlyielcreeoyeiao)aeaecfd,(dzlntbinfatNeeceveaonstdaddATaTsrd=eordnfoCanmrhhon3nrPidstfstnob3eeiro)ioFeaoinn(esnrulmaOcPphnpsgdcehrnoPai<Oolef,SiagoudafnnfnFr0wlar,SnelsftfedeOo.na4rlciwieF0uo1ofmtd.rhtadS5i5isaoa0lnotsci)t7rtss0rn:atafhndsePothbo0onno<Tee+aoFteeorittlto1ph-cmxceaoOdeuflx%peltrmp0udgyiipaaSminmn.oni4cnl7oa.wcditssgii-eco0unatnieTfhseynasnolida,sohiim1lgptrtgegusmue4itsppvcohonlao4r.faaermdlanitfoLsethfpvtla.ThgPinaueoeCetyc(oabFhanoaruraNsi5ltfoatelOrnnoeilwe0a=glsoutgairS.fole(3aznoaoymtyPvOs)eetrip.f,eoFodernogcvePdrrOfcuacoresetOttctinoetnerSsahhnhesntprSdn)eeePsenuieouFedagiFmtlc41sncfatihOtiiuw4vtgPisotehtovleSehFednfduognreoeiOa,rourlgaouniyydelurnfnhSsfuyaa.rdeitiirneyltinnnTeisen(xngffghhcprhe1owlpsoaauu<.et4,u3olhlveors0Aeanbspiareemrd.tundleonirn0iarddeoyasi5Pdpecmeanlt1)c.Oodpre-t0eod(aaleeedAS0lrPslolrrxeuaile0sFti,OsnepctornnskewictdpodSitneeintipsf.wdFedegcicoumnrdtnOa)rerrhetootee.hrtaroeenc.hpyeo,getovestbshada3anheyresnaaesn.rta,nre0dreyncudtwoors8roivmePuzmlgbveeremOh(ri+euidrgaoatnroSm-iih}rtao1frnFh'to-tln.sower6nt0ahse Iab.pepursoexdiTSmahtsauitsgdeuystithduOeadbny4jc-ewehcoatfiousvrrdeLao: Cn1e530-towveeaeslktuaebinloihsfahPlaOdtaiSotaFn.osTntuhadecyur.etesuilnthsaolafttihoins troaxnigceitfyinadnedrtsotudy will IAT7I0#..98Id.3enTtiefisctaAtiPNortOniac:mSleFes: CStgruFcptSuOrejF 5M0W2 (g/mole) 1D.6e9nsity (g/ml) B. PlAch1a9hiuFbngara--hrifNltaryypcaMtutcaiecrtnRriiaidtolzySneSLapftoaateaebrscbdmotimrlraoiaoatnsytnfcdo:ytorhaRypiec/meyisnSdpwpo/MuwomarnDsaiisttncyeitoaborclniwloddPimteauOytscpehSotroeeFmfnddrepi.enbnrsCeyoteshdat3heuarMmceschtppiSc(cou1aplsri)lsiee.ticcnybhitla.oaelrT.tfaytThchAetheeedPrnihOszoeaamSsmtiFiivopnenslaesalbmw&pypraoC'lsHedhfu-hoeNcamutsMniadbcnRaedtleosantnobde a C. Htemanpdelrinatguraen.d Storage: Upon receipt, the test article was stored tightly sealed at room D. rDeitsuprnoesidtitoonthoef TreessetaMrcahtecrliiaeln:tAanftyerrecmomaipnlientgiounnoufsethdepsoturtdioyn. of the test article was III. Test System: 0 l5 2 6 2 TFi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test ggrf(al(leotatnaewsresgwirnaeeagttxseMpcdacootaabsnl3lueycc.r0euremnalLattcisrtx/haemiwatdnimioegnbrnbaeti)eesn(resetMsotxdcp(Wto3ohonmes)r/eau2cpdts4hsoi/t.anuco7mgnh9tdaeb)thmsexfterrboc(aIelmodngrme/d/caa1poel0somxGy6uhrpafnaiiosndugug)sLinetnxaedpwd)a(u.aitremhiTrqfrfhpouoleorauiwntgf4iltohoohrnwaoa:atu-evtsr)ehtsrnrxeoftao.(um1rTg/ohhdoneeafenstfsmydrieratoyisynsh)pgixnhaeie(rf1r1l3eo0-w0wli0taesr pl/mL) tGMca1aoi/irrdWnvgfeleveonnet=ws:rcistgoiryo/an:mtnce(eom:fnla(etLcLr)t/ao/gtmri):oinn(1)(pgpramm))/(l x 106pg) conversion factor: 1000 fil/mL 1 DTairroefsaleotewGd:TraorhPueeOpptSarForgveidt ecdoninc1C(ep0tnh0pte0mraf)toiollnoswoinfge3S(atpa0cylbhr/ilmnete:gisnet)FcolomwpoRuanted, the syringe flow' 2A(L.i0/rmf-l3oin.w0) Rate rate and the Control: Air only 2.0 - 3.0 FfdgvlceooeaoovrnnylueucderlzmoirieeoatnirertfocoeSTcne1dtoonrsa0abntfn0arnyttd0asahsataiftpeirnhoodpdiserndnmmttsiuPgan.dorOgItCgyhnfSe.aofettFreOncaasctarndntroelooiccnddmlsoucaeaSmaleyanlnpgstpziteer.mroeodcaAruutatoranipilo,omrsdfmnaawloannsoeuniewtfmandrroteytmwraoole(exasoFxfa.syspTdcpgiohiInheeRscnecet)rroed.(eems2aCtto)osp.hmeforTadaoaumhptowneepbmdrhredioeawraapnlyaiertrniirooraeuetfwcanepeddartcsoeeessosspramntiaenrccryqogeehundntawioitmvtaroamaanbsrltdeeeiadodnriaenntonbstciwaafyghilkannyFansazmTaoteeIwbdRdenbry tihhmuemm3aeMndeiAaLlytnAeileRmyuCtpahl.rasiBonwriozetdeorydeewbhuyaethniCgadhnOleats2dsiawaisn.epArahecytcdxtoheiratedetitraoemnrncmineaienwnddaititnimhoecanmrnooeadpfnistahiiemteesalsywltuupedsrryeiao,gpreaetlroplfraodonrotmiosmcienoadgll.saawpnpderroeved by pwzvsSteraeaaerlrnsoisuddspmfauaertrrorewedfmdoal(ireTsqrmxaHuctteoirsPddalFolcecatOfchictotcrShenoo)edmrpwdlariaimtaontsoncgiegetadtcrdtoaerudopsrpetephudgsyb.(yr/4ltoi)afsu,onhpwrde.ePidUtmFhmrOiaeneSltseehlcaiPogtndrFhaosOtlsy(mpS3sri)oasl.edy.UviAfmerinlicasnaasewltsyiwoessnripaseesiocndctferottohPlelsaFrectmOcontiSpeonydelsed(uovHrvferoePolrslLglnobaCiwgty-ehMihntigSogn/hMa-dSay) 001527 3 T-7098.3 Acute POSF Inhalation Limit Test. Final Report fpferovloarammclueadatlhtiidenoLinrlsuieo.mnclgtuiltstyi,toeilnnsivttfoegorrpr,orksuouippdb.ynsAleeeyqnp,ueobetrnluattibdohednisesotrfoifalloleneadgdcibchwartaliitisehnsvulwaieqleuuwraeitadiscononiiml.tlreTomcghteeeeddniraeaftmoternlayepincofirisnxosgepibdsslyeeincoftnuaiotud1nar0sey%w1e4re IV. Results and Discussion: TttfnccahooooerernntsgttccfbehaeeAteecnnctttttlmoThrritamahnehottcaeiissioetotapPnnntmthheOt.erieseanrSt.etcFgitTaohrlhanoecetuumffplolaaooir(trwse5tfphld)-oh.ethwceTlroriohemwnuecgaigemteshnnteietecnrasahrcstataruigtemoerraednobsdueebipndcras.osttFhneowdecah3eceoentlxonnitptrretatsohhrtswsieeou/hnIsrmdeyicieisnchnatheultmmahtGemeasarybcestheaoaLcracaahmcadweovbjsueuearnseranttqgwdcfueohatidlhrsaiebmt99hr11abii2usc%etmriunaoc9mlfl1utahdypeepm ControlTGhreotuhpr:ee control group male rats had no adverse clinical observations noted dinuirtiinatgiotnheo4f thhreasitru-odny.lyTehxepcoosnutrreo.lTghreouapvegraaigneedboadnyawveeriagghet owfa2s237g8(5.51%2)goaftbthoedy "Weight during the 14 day study period. Liver weights averaged 14.1 1.0 g and the cwaaecSvhnxhueeadtarlrrngnlatagigrivgcneeeaaeasssNlautaleisafientdvsfniclooedlargucnrotriHentpoaoodgstiuf.essybptbdOooasfl)nipowdtnhdaohyidetdtauhiwrhdnroategiChlmrniosoaOgaigunahrl2yndyntteddhrStbuoaeaeltttsahhrrimedovteaedionarhwcerdiaeiotrasesislifstr,ieaoanasi0.cptsea.taa0sIflnt,tlc3ohdowth5uohninblasrgodresteooge0insnly.notoo0cirg0toAhfeei1nitpcdpohaopo(letlerTehftscanaaa.obntdblnTyvaisltxleehorDyoloims)1srll.i.i.sesw.AwEroalleifsdltvhterpehanoaoetlscLellscuudaianbnmsnglgiypoossrna(sewabnicgleohanrtteghi,fon3icncMooaatnnlttrol POSF Limit Grout?: rPptbtcadlhheeosooeOvecetndsceoeSttuyo1rlivFesrmnm4weixu-notrippdreoTyDnefeaoaiuahagPparywstseetyhuFeileedosyrtrtOdtrzinheurteoefS..oraadodretd.TnToyriceooiaE,hhomfpnfPeooeanwteeOaecvutrlararlhieiwrSvosmosreheluFindh.x0tiorri.ahtap.Tiggu0dnLggorhreh3eroessiaate5ov.uuncsuaueaTrpahmreprivmihnmvnew0ofegcpaero.uraare0allerelnuieisg0engatatewe2hweshmroeecdeabt(afsaprtTosPoasePlladacnOefpFwvnybusurSOaoelleerwweavrFripSbatneeee2aegitrerioirrd)eaegenee,lxagdhodafsncpuerttigood1oremiiwoinl4mdscnlfife.eaaelde2gdaclst1culhrwrt9ey4opteesotgad0ulo2siaso4pc9(i.av0cg4.ua13noegn.tAt2.9depr0ni3a.a%fsu8tei1grnIctaox)ned9aogltdg1osnront1ae.rh(tffopNcp.deb6etspteahf0oesam=nfyeddvepod3,yocedpbPe)ttrnunmsowsOafeeptogoemSrludpaeviivFgncrioeaelltadhasuitermvtitsteteo-tauieehnedonrraxe1un,edstp4rsotuaitho-nhrydoesegesafudyre average percentage of PFOS in urine was 0.05 0.03% (Table 3). The standard curve 001528 4 FTi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test tuhseedarteoacuanlcduelrattheethceurlevvee(lAs UofCP)FfOorSthineseexetrxatcratecdteudriunreinseamstapnledsarisdsshaorewinniTnaFbilgeu4r.e 1, and c0t0fooo..00xun00incc47deo%nmkitnTiSrgn(aehtT.ethreitTaueoisbcmhnesleeesfostriau5fnauvd)mdmP.eyiFrpnaaOolggenfsSsedPaw(puOrTeereSiarrnscbFeieelmcne((oTti45all-)al)g7.e.re0cTt9tooeh8fdte.hi4aane)vthPienanrFleaeOwcgdrSheoPipcaOlsehmyvSaoeFolusnsinmnodtabasolesylferpPur1vemF3reOcpdaeoStnisnnittan-eadgcnsoereosaroepcufscmtyoPoFmwwmOpaaeSassans00wyu..01irane05sg18tahleso V. Conclusion: The apparent inhalation 4-hour LC50 in rats for POSF is greater than 1000 ppm. v 001529 TFi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test VI. References: 1. KLSpaebesctontrreaorts,ocrToy.p.CyR.hee3qmMueicsSatplNeCcohi.aa6lrt3ay7cMt0e.raMitzeaartiyiaol1sn5&o, 2fM0P0Oa1nS.uFfa(cFtMur-in3g27D0i)vbisyio'Hn -ANnMalRyti&calI9F-NMR 2. Modified EPA Method 320. Pace Analytical. (2001). 3. MHSpaaentcrsiiecfniec,s,.KQE.Jun.av,niCrtoiltenam.tiSvecnei,.CLah.naAdra.T,cetEeclrhliez..fa3sto5ion, n,7M6o6f.-O7E7.r,0gJ.aonhicnsFolnu,oHro.cOh.e(m20ic0a1ls).iCnoBmioploougnicda-l 4. MoeltehecethtrroofdlsupNorrauoymc/mhbeeamrs:sicEsaTpleScc-ot8rm-o9pm6o.eu0tr.nydE/smxfatrrsoasmcstipuoernicntoreoffmpooerttaarsyns.ailu3ymMsispEuenrsvfilniurgoonHromPoeLcntCatan-lesLualbfoonraattoeryo.r 5. IaGnnucdt.zT3kMeocwhL,naoTbl.oo(gr2ai0etos0.r1y) RPOeqSuFesAt Nniom.aEl0E1x-p0o6s4u8r.e3TMesEt TR&epSoSrt.FiPealcdeAAnnaalylytitcicaallSSeervrviciceess, 6. PSeefalcuaot,ocAt.anaensduTlfuornnyelr,FTlu.o(r2id0e01()PAOcSuFt)eTIn-7h0a9la8t.i4o.n Toxicokinetic Study of 001530 6 TFi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test VL Signature Page Authors: ATStonuxddiryceowDloiSrgeeicsattcoaSrtp,ecPiha.lDis.t Reviewed by: JTSotouhxdniycLoD.loBirgeuictsettonSrhpoefcfi,alPishtD, CIH, DABT Date Date 001531 7 Table 1: Control Group Biological Data Exposur animal number 5/1/01 Animal weight Pre exposure Day 0 (g) 5/2/01 Animal weight Post exposure Day 1 (g) 5/3/01 Animal weight Post exposure Day 2 (g) 5/7/01 Animal weight Post exposure Day 6 (g) 5/14/01 Animal weight (g) T-Test p-value Predose vs day 6 BW (1) 5/14/01 Necropsy Liver weights (9) Lhrer/BW ratio Control 1 R01462 374 372 369.1 376 391 0.04 14.1 0.036 Control 2 R01463 368 364.5 360.1 373 386 13.1 0.034 Control 3 R01464 392 376 386 401 422 15 0.036 avg 378 371 372 383 400 14.1 0.035 SD 12 6 13 15 20 1 0.001 (1) p-value for a 2-tailed Student's T-test, assuming equal variance. Predose vs. day 6 body weight. Statistically significant if P<0.05. notes (overnight fast) FTi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test. Table 2: POSF Limit Dose Group Liver Weights Exposure animal POSF number (ppm) 1000 R01465 5/1/01 5/2/01 Animal 5/3/01 5/7/01 5/14/01 T-Test Liver weights Liver/BW ratio Animal weight Post Animal Animal Animal p-value (9) weight Pre exposure Day weight Post weight Post weight (g) Prexpo 5/14/01 exposure i( g ) Day 0 (g) exposure Day 2 (g) exposure Day 6(g ) vs day Necropsy 6 BW (1) 374 372 370.7 379 390 0.01 12.8 0.033 1000 R01466 364 355 360.7 368 383 13.6 0.036 1000 R01467 416 392.6 409.8 429 440 16.1 0.037 avg 385 373 380 392 404 14.2 0.035 SD 28 19 26 33 31 2 0.002 P-value 0.72 0.85 0.63 0.70 0.84 0.93 0.880 0 _______ ((12)) pp--vvaalulueefcoormap2a-rtainilgedcoSntturdoel nvtss.'tTre-atetesdt, gasrosuump ibnogdyeqwuaeligvhatrsi.aSnctaet.isPtirceadlolysesivgsn. ibfiocdayntwiefipg<h0t.o0n5 day 6. Statistically significant if p<0.05 001533 9 TFi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test. Table 3: Limit Group PFOS In Urine Area under Concentratio the curve n of PFOS In (AUC) urine (ppb) Samples MeOH Solvent Blank R01645 d1PD R01645 d1PD avg SO R01646d1PD R01646 d1PD avg SD R01647 dIPD R01647 d1PD avg SD Overall AVG Overall SD Abundance xm in 8888 1154557 949883 1052220 144726 394606 460644 427625 46696 1384246 1238595 1311420.5 102991 (PPb) 94 1936 1573 1755 256 590 707 648 83 2343 2085 2214 182 ppm PFOS 1.94 1.57 2 0.26 0.59 0.71 1 0.08 2.34 2.08 2 0.18 Dilution factor Collected Amount Average Amount of PFOSin Overnight Urine Average Undiluted urine PFOS (ppm) Volume of Urine (ml) Total PFOS (ug) Total PFOS (mg) % PFOS in urine (3) 2 3.88 2 3.14 3.51 10 35.1 0.62 0.04 2 1.18 2 1.42 1.3 17 22.1 0.17 0.02 2 4.68 2 4.16 4.42 18 79.56 0.37 0.08 3.08 1.60 0.05 .-0.03 0.06 0.04 0.14 0.08 0.05 (1) Standard curve equation where Y= area under the curve (AUC) in units of relative abundance multiplied by time (m inutes), and X = ppb of PFOS in urine. X = (Y-61687)/564.53 (2) Mean + SD of PFOS in urine are shown for N=3, where each of the 3 values are the mear> of duplicate determinations. (3) Inhaled Dose of POSF: (2.0 ml/breath) x (120 breaths/min) x (1 mg POSF/ L of air) x (240 minVK Body weight day 0 (g)/(1000g/Kg)______ 001534 10 FTi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test. Table 4: Extracted Urine Combined Standard Curve for PFOS Sample MeOH Solvent Blank Milli Q water Control urine Matrix blank MS 4 ppb MS 8 ppb MS 20 ppb MS 40 ppb MS 80 ppb MS 202 ppb MS 404 ppb MS 808 ppb Control urine Matrix blank MS 4 ppb .-MS 8 ppb MS 20 ppb MS 4 0 ppb MS 80 ppb MS 202 ppb MS 404 ppb MS 808 ppb POSF AUC (Ml) 11952 36312 35330 32394 , 53129 56366 77325 119932 147773 220143 442924 45441 71331 54846 70255 63918 197266 304118 293452 592511 001535 FTi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test. Doaa (pp m ) A nim al* Predose Body weigh! D ay 0 (g) Inhaled Dose P O S F (1) (m g/kg) Table 5: POSF Limit Dose Group Serum PFOS Data Body weigh! day 1 post doss (g) Body weight day Body weight day B .W . day 5 13 post dose (at 2 post dose post dose necropsy) (g) (g) (g) Serum volume (2) (ml) , serum ppm (PFOS) (3) (ug /m l) Amount ot P FO S In serum (4) (m g) Available P O S F In Inhaled air (5) (m g) % o( inhaled P O S F In serum as PFO S at necropsy serum) 1000 ppm POSF 1000 ppm POSF 1000 ppm POSF Avg STD R01645 R01646 R01647 374 364 416 386 26 154.01 156.24 136.46 150.24 10.42 372 355 392.6 373 If 370.7 360.7 409.8 380 20 379 368 429 392 32.6 390 383 440 411.6 40 12.02 12.37 14.21 12.87 1.18 5.1 48 3.8 4.57 0.7 0.061 0.059 0.054 0.068 0.004 57.60 57.60 57.60 67.60 0.00 0.11 0.10 0.09 0.10 0.01 (1) Inhaled Dose of POSF: (2.0 ml/breath) x (120 breaths/min) x (1 mg POSF/ 1000ml air) x (240 min)/(( Body weight day 0 (g)/(1000g/Kg)] (2) Given: (32.3 mis serum)/(kg of body weight). Serum volume =(BW day 13 (g)/1000g/Kg) x 32.3 ml/Kg (3) Serum ppm: Analytical work done by Dave Ehresman PFOS in serum measured by HPLC/MS (see appendix 2). (4) PFOS in serum (mg): ((Serum volume ml )(Serum PFOS ug/ml)] /1000 ug/mg (5) POSF in inhaled air (mg): (2.0 ml/breath) x (120 breaths/min) x (1 mg/ 1000ml) x (240 min) = 57.6 mg POSF in inhaled air (6) % inhaled POSF in serum: [PFOS in serum (mg)/POSF in inhaled air (mg)]*100 001536 I2 TFi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test. 700000 600000 500000 soI<D 400000 300000 200000 100000 0 Figure 1: extracted urine combined standard curve for PFOS Senesi ------ Linear (Seriesl) 100 200 300 400 500 600 700 800 900 ppb PFOS in urine 13 00153V FTi-n7a0l9R8.e3poArctute POSF Inhalation Limit Test. Appendix 1 Histopatholoev Results Necropsy 5/3/01 Abrlolnrcahtsiolulensg.sThhaisveisgpeonsesriablliyzeadn magioldnatol cmhaondgeeraatesscoocniagteesdtiwonithofCa0lv2eoeluithwainthasoiac.caSsoiomnealluenxgtrsawvaesraetinoontowfelblloinosdufafrloatuendd. arterioles and NACoonnimstirgaonlli#Rfi0ca1n4t6c2h:anges in lung and bladder. NTooxsiicgonkiifniceatinctRch0a1n4g6e8s:in lung and bladder. TOsiognxneiicmfiocakacinrnotesctcihcoapnRigc0e1foi4sc7pu1rse:osefnatteilsebcltaadsidse(rc. ollapsed alveoli) is present in one lung lobe. The change was chronic in duration. No NTooxsiicgonkiifniceatinct cRh0a1n4g7es2:in lung and bladder ; 001S38 14 SAerpupmenPdFixO2S To: Andrew Seacat/US-Corporate/3M/US@3M-Corporate Tommie Tumer/US-Corporate/3M/US cc: John L. Butenhoff/US-Corporate/3M/US@3M-Corporate Paul Lieder/US-Corporate/3M/US@3M-Corporate Sue Tanaka/US-Corporate/3M/U S@3M-Corporate Kathy Thompson/US-Corporate/3M/US Deanna J. Luebker/US-Corporate/3M/US@3M-Corporate Subject: PFOS Levels from rats exposed to POSF via inhalation T-7098.3 Andrew and Tommie: The 1 to 100 estimate for the serum levels of PFOS was right on target The following results are based on the rat serum being diluted 1:100 prior to extraction. The extraction was carried out at pH 3.0 using a single shake out in Ethyl Acetate. The EA was transferred to a clean polypropylene tube and blown to dryness using nitrogen gas. The resultant residue was re-dissolved in 100 uL of 25% acetonitrile and 75% 10 mm ammonium acetate. Of this solution 20 uL were injected onto a Betasil C18 reversed phase HPLC column (50 x 2 mm, 5 micron particle size). The flow rate was maintained at 0.5 ml/min. A gradient flow of acetonitrile was used to elute the compounds of interest starting with 15% acetonitrile and ending with 65% acetonitrile over 5.5 minutes. Return to initial operational parameters and cdtumn equilibration requires and additional 2.5 minutes for a run time cycle of 8.0 minutes per run. The MS was operated in the Electro Spray negative ionization mode with a constant 3.0 kV potential applied to the source. The method used was an MS/MS method involving the transition of the following ions: Compound Q-2 Offset Voltage Base Peak MS/MS Ion Internal Standard (PFHS) 60 volts 299 amu 80 amu 60 volts PFOS 499 amu 80 amu The capillary heater was held at a constant 300 degrees C. All quantitative calculations were based on the MS/MS ion rations between the compound of interest (PFOS) and the internal standard (PFHS). The standard curve ranged from 5 ng/mL (ppb) to 250 ng/mL (ppb). A quadratic curve fit was used with the standard value weighted 1/X . This curve fit produced an R squared value of 0.9935. The first tube of two available for control animal number 1R01464 was diluted and run with the initial extractions. This tube (1:100 dilution) had a PFOS level of 2.3 ppm. Repeat of this tube dilution produced comparable results (2.1 ppm). The second tube labeled 1R01464 was diluted 1:100 and extracted. The second tube on this control animal had a result of less than the lowest standard (5.0 ppb on the diluted serum). 001539 FTi-n7a0l9R8.e3poArctute POSF Inhalation Limit TesL Only one serum tube was available for animal 1R01465, unfortunately this tube was labeled 1R0465 with the correct date and T # recorded. I diluted and extracted this tube and recorded results under the animal number 1R01465. ~7 The sample for 1R01468 was noted as hemolyzed (2+ hemolysis). The initial result of ~1R01472Tappeared to be significantly lower than 1R01468 and 1R 01471. This sample was also re-extracted and the analysis repeated. No significant difference was noted on reextraction. Results: Rat # Control PFOS Level 1R01462 1R01463 1R01464 yes yes yes <5.0 <5.0 <5.0 ppb (on Mdiluted serum) II 1R01465 1R01466 1R01467 1R01468 1R01471 1R01472 no no no no nono 5.1 ppm 4.8 ppm 3.8 ppm 8.6 ppm 9.3 ppm 5.0 ppm (5.1 ppm on duplicate) dje David J. Ehresman 3M Corporate Toxicology Bldg. 236-1B-22 Phone 651-733-5070 FAX 651-737-4754 djehresman@mmm.com UU154U 16 TFi-n7a0l9R83epoArctute POSF inhalation lim it Test UU1541 17