Document N2QQJRgLrNn2kZkRb92kdmrvw

3M Environmental Laboratory Final Report- Analytical Study Single-Dose Intravenous Pharmacokinetic Study of T-6052 in Rabbits In-Vivo Study Reference Number: HWI#6329-134 TSetsutdySuNbusmtbanecre:: FACM-D1T2-011(T1-669045.2)1 Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facilit3yM: Environmental Technology & Services 935 Bush Avenue St. Paul, MN 55106 Method Numbers AMDT-M-1-0, and Revisions: DThoehrrmmaalnEnxtDrXa2ct0i0o0n oOfrgFalunoirciHdaelbiydeMAenaanlsyozfera-LMiovdeirfied AMDT-M-2-0, Fluoride Measurement by Means of an Orion EA940 Expandable Ton Analyzer AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver AMDT-M-5-0, Analysis of Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry AMDT-M-8-0, Analysis of Fluoride Using the Skalar Segmented Flow Analyzer With Ion Selective Electrode AMDT-M-14-0, TDhoehrrmmaalnEnxtDrXa2ct0i0o0n oOfrgFalnuoirciHdaelbiydeMAenaanlsyzoefar-SMeorduimfied Initiation Date: See attached protocol Author: James D. Johnson Approved By: her Sjtaumdeys DDi.reJcfhohrson sre Completion Date 600802 1 T(Th-e60l5iv2e)r wsearmpelaensaaltyz4e8dhboyurcsomafbtuesrtsiionnglfeoirnttortaalveonroguasniadcmfilnuiosrtirnaet.ioTn-o6f05F2C-i1s 2a0 0.02% ug/kg) solution samples hoafdFCd-et1e2c0t.ablOenlorygtahneic2f0l0uomrign/ek:g4(81a0nudg/1k0g6) uagn/dwh1o0l0e0 lmivge/r,kg (50 respectively. doses. There iTshaesmeartrkievrialfoarmdoeurnmtasloafbsoorrgpatniiocnfsltuuodriiense, are a if the reflection doses are of the low higher than used in this study. cTohemploivuenrdanisdpseerfrluumorsoadmepclaensesfurlofmonHaWteI#(6a3m2m9o-n1i3u4mwsealrte).avTahielarbeleisfnoortaenaxlpyescits,edTthois be are aenxypebcitoetdratnosfboermsaitmiiloanrotfo tthhiastcfoomupnodufnodr paenrdfltuhoeropohcatramnaecsouklifnoneattiec.s and The disposition tissues at m4a8shsousrpsecwterroemeatnraylyfozredpebryflcuoomrboudsetcainoensuflorfotnoattale oarngiaonn.icTfhlueordiantea awnedrebytoelbeectrospray analyzed The high tdoosperoivsijduestda5t0a for the ug/kg. aTs-s6e0s5s2meinstao0af.0s2u%bsseolquuteinotn doefrFmCa-l1a2b0s,orwphtiicohn study. is 25% solids. 3T 0 ESTMAT ~~E ~RIALS 3.1 Test, Control, and 3.1.1 Analytical Reference Reference Substances Substance: and Matrices FC-95, lot 161 or 171. They are equivalent. 3.1.2 3.1.3 Analytical Analytical Reference Matrix: Bovine Control Substance: None liver and bovine serum 3.1.4 Analytical Control Matrix: Bovine liver and bovine serum g3r.2ocSeoruyrscteoreo,fbMoavtienreiaslesr:u3mMfrIoCmPS/iPgCmPaDCihviesmiiocnalfoCroFmCp-9a5n,ybovine liver from 3.3. Purity and Strength of Reference Substance: Responsibility of Sponsor. 3.4 Stability of Reference Substance: To be determined by Sponsor. b3i.o5lSogtiocraalgseaCmoplnedsittihoensstofroargTeeisst-2M0a+t1e0riaCl.s: Room temperature for FC-95. For 600803 2 s3Gt.Lu6dPDyiRsiespgoiusnliaptatirioaonlnloeflforwSipttehheciatmi2em8nesdp:aeyrBiaiobodslororegpqituciiaorlnetdsitsfsuoudreyss,tausdnoidealsflllutoiidnsssguewerisltlwhiablnel 2br8eetrdaeaitynasei,dnepTdeh.ris 40EXPERIMENTAL-Overview Sfoerraunmalaynsidstfiosrsufelsuofrrionme acnoimmpaolusnddoss.eSdianscedepsecrrfilbueodro(dHeWcIa#n6e3su2l9f-o1n3a4t)e,anwieorneiasvpaioltable abniaoltyrsainssffoorrmfeldu,ortihnee.anaTlhyesifslwuoarsinaecdcaotmaplaireshreedlawtietdhdicroemcbtulysttioopnerafnlduosruobdseecqauneen-t msualsfsonsapteectanrioomnetcroyncpernotvriadteisone.viAddednicteiotnhaalt atnhaelpyesrifslouforloivdeercasnaemspullefsonwaittehaenlieocntriosspray parbessoernptt.ioDnaitnaafsruobmstehqeuseentansatluydsyis(wHiWlIl#b6e32u9s-e1d3t5o)a.ssess the extent of dermal 5.0 EXPERIMENTAL - METHODS 5D.o1hrAmMaDnTn-MD-X12-000,0 TOhregranmiacl HEaxltiradcetAinoanloyfzeFlru-oLriivdeer by Means of a Modified E5.x2paAnMdDabTl-eM-Io2n-A0n,alFylzuoerride Measurement by Meansof an Orion EA940 5.3 AMDT-M-4-0, ExtractionofFluorochemicals from Rabbit Liver 5E.l4ecAtrMoDsTpr-aMy-M5a-s0s,SpAencatlyrsoimsetorfyRabbit Liver Extract for Fluorochemicals Using 5.5 AMDT-M-8-0, Analyzer With Ion SeAlneacltyisviesEolefcFtlruoodreide Using the Skalar Segmented Flow 5D.o6hArMmaDnTn-MD-X1240-000,OTrhgeanrimcalHaElxitdreacAtniaonlyozfeFrl-uSoerriudme by Meansof a Modified `liTvheerdaatta48(Shkoaulrasr)afitseartatnacihnetdrafovrencooumsbduossteioonfaFnCa-ly1s2i0s.waTshenotnotdaelteocrtgeadnifcorfltuhoerine in control, 0.1 ug/kg, and 1.0 ug/kg groups. The total organic fluorine measured for 600804 Ethleec1t0rousgp/rkayg maansds5s0puegc/tkrogmreatbrbyit(sseweeraett4ac8headn)dc1o0n6fiurgm/ewdhothleeplrievesre,ncreesopefctively. perfluorodecanesulfonate (m/z=599). aOntahleyrzedart(aliwvaers acnodllseecrtuemd)u,siSnkgaltahre sDeogrmhemntaendofrlgoawniacnahlaylzideer analyzer, with ion Orion ion selective dealteac,traoldteho(suegrhusmu)p,poarntdiveel,ecitnrotshperoapyimnaisonsofsptehcetrSotmeutdryyD(ilrievcert)or- see appendices. This is not required to reach the conclusion stated here and therefore is not discussed in detail. 6.1 Circumstances that May Have Affected the Quality of the Data: The problem with this analysis is that there is not these intravenous doses because the doses are nearly enough too low. fluorine in the livear fter `This pharmacokinetic study is not useful in terms of providing data for the eaxspseecstsemdenits onofta odbesremravledabisnorlpivteiroenxscteupdty.foTrhaetpraecreflautotrhoedehciagnheessutldfoonsiec(a5c0iudga/nkig)o.n If the dermal absorption study doses are high enough, there is a marker. 8.0 MAINTENANCEOFRAWDATAANDRECORDS 8.1 Raw Data and Data: appropriate specimens, and Raw data, electronic approved protocol, approved data will be maintained in the final report, AMDT archives. 9.1 Protocol and Amendments 9.1.1 Protocol and Final Report: HWI#6329-134, "Single-Dose Intravenous Pharmacokinetic Study of T-6052 in Rabbits" (Protocol type TP8084.PK for dosing of animals, tissue collection, etc.) 9.2 Si9g.n1e.d2 RAenpaloyrttiscaflrpormotIoncdoilvAidMuDaTl-S1c1ie1n6t9is4t.s1: None 9.3 Quality Assurance Unit Statement: See attached 606305 4 9.2 Signed Reports from Individual Scientists: None 9.3 Quality Assurance Unit Statement: See attached 9.4 Key Personnel Involved in the Study: See attached 9.5 Materials and Equipment: See methods 9.6 Solutions, Reagents, and Standards: See methods 9.7 Sample Preparation: Sce methods 9.8 Quality Control Practices: See methods 9.9 Test Methods: See Protocol AMDT-111694.1 9.10 Instrument Settings: See methods 9.11 Data: See attached. 9ex.t1r1a.c1tiSounmfmoalrloyweadndbyraawnadlaytsai;s uusginFg" iOnriwohnolieonlaivnearlyazserd.etermined by thermal 9.11.2 Summary and raw data; analysisofliver extracts using electrospray mass spectrometry. 9.11.3 Summary and raw data; extraction followed by analysis ug F" using in whole liver as determined by Skalar segmented flow analyzer thermal with ion selective electrode. 9ex.t1r1a.c4tiSounmfmoalrloyweadndbyraawnadlaytsai;s ppm F" in serum using Orion ion as determined analyzer. by thermal e9x.t1r1a.c5tiSounmfmoalrloyweadndbyraawnadlaytsai;s ppm using FS"kianlasrerseugmmaesntdeetdefrlmoiwneadnablyyztehrerwmiatlh ion selective electrode. 606306 3 9.1.1 Final Report: HWI#6329-134, "Single-Dose RIanbtbriavtesn"ou(sPrPohtaorcmoalctoykpieneTtPi8c0S8t4u.dPyKoffoTr-d6o0s5i2nginof animals, tissue collection, etc.) 606507 ? MPWAODSITISSOOFNCFIWiCOE Ns3B7OS0X7.I7755N4458 Sponsor: St. Paul,3mMinnesota + CORNING Corry FINAL REPORT Study Title: Single-SDtousdey IonftrTa-v6e0n5o2usinPhRaarbmbaictoskinetic Author: Steven M. Glaza Study Completion Date: February 1, 1995 PerformingLaboratory: H3a3z0l1etKoinnsWmiasnconBsoiunl,evaIrndc. Madison, Wisconsin 53704 LabPo rojr ectaIdt entio ficr atiy on: HUI 6329-134 Prone sos2arieen EXPRESS'MAIL OELIVERY Page 1 of 24 3301 KINSMAN BLVD. fax 00s MADISON. wi GOCBO8 53704 Page 2 of 24 QUALITY ASSURANCE STATEMENT Hyha This report has been reviewed by the Quality Assurance Unit of Hazleton W(iFOsAc)onsGiono,d LIancb.o,rationryacPcroradcatniccee wRietghulatthieonFso,od21anCdFRDru5g3.5A5dmi(nb)ist(r6)ati(o3n). STthuedyfoDlilroewcitngor iannsdpemcatniaognesmewnetr.e cWornidtutcetnedstaantdusfinrdeipnogrsts roefporitnesdpecttoiotnhse SatndandfairnddinogpseraatreingispsruoedcedtuoreHsa.zleton management monthly according to IFnrsopnection DTaotes ____ Phase Date StuRedpyoDrtierdecttoor MaDnaatgeemteont 11/05/94 11/11/94 0011//3100//9955 11/05/94 11/11/94 0011//3100//9955 Protocol Review Animal Observation RDeaptoar/tRepRoerrteviReewview 11/08/94 11/11/94 0011//1300//9955 12/10/94 12/10/94 0022//1100//9955 u [ / Lf Kn . ) ) ReprielsieantMa.tiDvaen,neQruality Assurance Unit 2 Jes Date 606809 Page 3 of 24 STUDY IDENTIFICATION Single-SDtousdey IonftrTa-v6e0n5o2usinPhRaarbmbaictoskinetic HUI 6329-134 Test Material Sponsor Sponsor's Representative Study Director Study Location StuEdxyperTiimmeenttaab]leStart Date Experimental Termination Date T-6052 TMoxicology Services S2t2.0-2Pa6u-l0,2 M3NM Ce55n1t4e4r John L. Butenhof, Pho T22o0x-i2cEo-l0o2gy3MSerCveincteesr S(t6.12)Pau7l3,3-1M9N62 55144 HSatzelveetnonM. WiGslcaoznasin, Inc. MPa.d0i.soBno,x W7I54553707-7545 (608) 241-7292 HBauzilleditnogn WNoi.sco3nsin, Inc. M3a8d0i2soPna,ckeWIrs A5v3e70n4ue NNoovveemmbbeerr 1131,, 11999944 06510 Page 4 of 24 AcuteToxicology Steven M. Glaza MSatnuadgyerDirector FStruadnycisCoo(rBdudi)natW.orMcDonald Patricia Padghan In-1ife Supervisor Rose M. Bridge Report Supervisor uality Assurance MSahneargreyrR. W. Petsel KEY PERSONNEL HWI 6329-134 LaboArnianaltMeodirciyne Supervisor Cindy J. Cary, DVM Diplomate, ACLAM Anatonical Pathology Jack Serfort/ DSuepbeorrvaihsoLr.s Pirkel Necropsy Anne Mosher Supervisor Pathology Data 6006811 Page 5 of 24 CONTENTS QSutauldiytyIdAesnstuirfainccaetioSntatement SKuemymaPreyrsonnel ORbejgeucltaitvoery Comp] fance TTeesstt SaynsdteComntrol Materials RPersocueldtusres DSiisgcnuastsuiroen Reference Table 21 IInnddiivviidduuaal] CBloidnyicWaelighStisgns(g) AppPerontdoicxolA TP8084.PK HUI 6329-134 Page 32 3 67 77 IH89 1 111 1132 114s 606812 Page 6 of 24 SUMMARY HHI 6329-134 aThdimsinissttuedryedwasbydoinnetratovenaosussessinjtheectiloenveltoofrabsbyisttse.mic exposure of T-6052 when t{Fhoeenmea/lvgeerhHoirucapl:)e.((NsZOtWne)rSiPDFlaeyrwa0,abtbeitrthsefoawrneirmeianljasescstirigeoncnee)diveoadrt aransdionmgleto inftirvaevegnroouusps injection 2, 20, 200, or 1,000 ng of of wTa-s6003.25/kmgl/okfgbofodry Gwreoiugphst 1(Gtrhoruposugh1 4tharnodugh1.053, mrle/skpgectfiovrelGyr)o.up `5The duse volune b4Ce8lfionhrioceuarlstesaotfbtsmeearrtveairtniitaorlnasveandwomeuirsneiscitonrnjadetcuitcoitonend.(DaatyBoad0p)yp.rwoexiAigmhbatltsoeoldwyers0ea.5m,pdleet2,er(m4a,ipnpe2rd4o,xjiumacanttdely ta4hneniLm)atlisnweasaotfc2o-le,xlpeec4rt-,iemde6n-ft,raol8n-,atner1m2a-iu,nraitcainuodlnar24(-4ah8ro-tuherorsuyrspoorsptom-saitrn-gjiienncjatelicotenia.orn)v,eIninaapdpodrfiotxtiioomena,telayt As2p0eppranorlxaitomefadtbelilonytood48wsaeshrouunorbstanadpi.onsectde-lilfnurjloeamcrteiafocrnha,ctaitnohinemsa,la.niamnadAllslsewnsetarmeptloeasntheeswtehSrepstonicszooenrdt.r wfiutghed. casobolbdlrieeucvntieapdte.endtobTgahrreobsiwsthaonlle,ecrTobiplvseeydr,evixbaaimlitenh,eatiaponondstbweaorstihornoktivdendneoanyesc,avfarh,aomweaveneadrc,hexsa&naiinsmgoauuliensawteweredev.e An pchoalslee.cted and sent frozen to the Sponsor after termination of the in-1fe Al five animals appeared normal throughout the study. 000813 Page 7 of 24) OBJECTIVE HNT 6329-134 Tthhee otebsjtecmtaitveerioafl,thTi-s60s5t2u,dywhweans taodmaisnsiesstseretdheaslevaelsinogflesysitnetmriacveneoxupsosuirnejecttoion to rabbits. REGULATORY COMPLIANCE TAhdimsinisstturdaytiwoans'sconGdouocdteLdaboirnataocrcyordParnaccetiwceithRegtuhleatU.iSo.nsFofoodr NaonndclDirnuigcal mLiaxbtourraetsoryforStucdoinecse,ntr2a1tiCoFnR, 58h,omwoigtehneitthye/seoxlcuebpitliiotny,thaatndansatlaybsiilsityofwatshenottest AcnoinmdaulcteWde.ifarAellAcptrocReedguulraetsiounsse.d inIntthhies osptiundiyonweroef tihnecSopmopnlsioarnceandwitshtudtyhe director, the study did not unnecessarily duplicate any previous work. TEST AND CONTROL MATERIALS Identification TThiequitde.st TmhaetercioanltrowlasmaitdeernitailfiweadsasSteTr-i6l0e52WaatnedrdefosrcriIbnejdectaisonc,leaUrS,P c(oAlbobortltess aLabcloeraart,oriceosl,orlLeostsNol.iqu8i6d-.748-0-02; Exp. March 1, 1996), and was described as Puar ndi Stat bily ity TdheeterSmpionnastoironasssu(miensclrudeisnpgonsuinbdierlittyestfocrontdeisttiomnast)e.rialA psaumrpiltey oafndthsetatbeisltity smattaebriilailt/yveahniacllyesesmixwtasurensot fotrakecnoncbeenftorraetiaodnm,inissotlruabtiiliotny,as hotmhiosgenweaistyn,ot and mraetqeureisatledwebryetchoensSipdonesroerd. toThbee apduerqiutayteandforsttahbeiliptuyrpoosfestheofUStPhisgrasdtuedyc.ontrol Stoanrd Ra etegntieon sTthoeretdesrtefmraitgereiraaltedw.as Asntyoreudnusaetdrotoemsttmeamtpeerriaatlurew.ill Thbee rcoentturronledmattoertihaelSwpaosnsor OapfetrearticnogmplPertoicoenduroef a(lSlOP)t.estiAnngy arcemcaoirndiinngg tvoehiHcalzeletmoany Wbiescuosnesdinfor(NoIt)herStandard testing and will not be discarded after issuance of the final report. 600814 Page 8 of 24 HHI 6329-134 Safety Precautions Tahned ptoelsitciaensd. control material handling procedures were according to HHI SOPs TEST SYSTEM Test Animal KAadlualmtazaolob,inMoicrhaibgbaintsonofOctthoebeHrra:5,(NZ1W99)4SPFandstmraaiinntawienreedraetceithveedHafzrloemtoHnRP, Inc., Wisconsin' facility at 3802 Packers Avenue, Madison, Wisconsin. Housing ADfutrienrgreaccecilpitm,atitohne aanndimatlhsrowuegrheoutaccthleimastteuddy,forthea apenriimoadlsowferaet Tienadsitvi7dudaalylsy. mhcaooiunnstteradoilnlienda scqtueramerpetener-rabsto.utrteonEonfvsitra1o9innmleetnsotsa2l3stCec,eolntarcoarlgeselsatfioivrnetthehemumpieadrniaittmayulreo-frooa5mn0d%wehr2u0em%i,dsiettayn-dtoa rc1eo2qn-ushiiordueerdre1ditgemhtopte/r1haa2vt-euhroehuardanddnaorhkuadm1viiedgrihstteyincegofnfcdeyicctltei.oonnstIhenexicsaststeeudsd,ywhotuehtreceyomwveea.rreiaAtdnioiocmnuasmlenftreodm tanhde fhoursbtahnedryCaraendanhdouUssiengoaftLHaHbIorcaotmoprlyiAendimwailtsh".standards outlined in the "Guide AnimalDiet TLhaeboarnaitomraylsRawbebreitprDioevtidHedF a#5c3c2e6s,s analyzed by the manufacturer for tPHoIwaFteeedrs,adIn1ci.bitTuhneanfdeeda miesasruoruetdinealmyount nutritional components and environmental of wceornetamnionaknntosw.n coSnatmapmliensanotfsthien twhaeterfeeadreorpewraitoedricaatllylevaenlaslyztehadtbwyouHlNdL.havTehere interfered with or affected the results of the study. SeleofcTetstiAnoimanls Tvheereansienlaelcstedweraet riadnednotmifibeadsedbyonanihneaa]lthnumabnderboadnyd wceoirgrhetspornedqiunigremeeanrtst.ag and 600815 Page 9 of 24 Study Destan HWI 6329-134 Female placed animals weighing from 2,813 to 3,031 into the following study groups: 9g at initiation of treatment were --Growp Treatment (1n-gD6os0e52Le/vkela) 1 (Control) 2 3 4 5 * T-6052 T-6052 T-6052 T-6052 0 2 20 200 1,000 * Sterile Water for Injection, USP. _Do(sne lV/okluam)e 0.5 0.5 0.5 0.5 1.02 ofNAunmibmearls 1 1 1 1 1 Nala jon for Si Sel, Historically, the New Zealand White albino rabbit has been the animal of choice because of the large amount of background information on this species. PROCEDURES DPosereparanadAdtminiistroatinon The test specific material was diluted with Sterile concentration for each dose level Water for in Groups Injection 2 Shrough to 4. achieve a The test material was administered undiluted at the 1,000 mg/kg dose level, using the bulk density of 0.98 g/mL to determine the dose volume. An individual dose of each respective test solution on its body weight on the day or of control was treatment. calculated for The respective each test animal based solution was administered by given as a slow intravenous injection into a push (approximately 30 to 60 marginal ear vein. The dose seconds in duration). The was prepared test solutions were stored at room temperature until administered. After administration, any remaining test solutions were discarded. ReasonforRouteofAdministration Intravenous injection is an acceptable route to assess systemic exposure. Observaotf Ainoimnalss Clinical 48 hours observations were conducted at after intravenous injection. approximately 0.5, 2, 4, 24, and Body weights were determined just before test material administration (Day 0). 600316 Page 10 of 24 Sample Collection HUI 6329-134 cAatbhleotoedriszaamtpiloen (oafpptrhoexiamuartiecluyla4rmLa)rtewrays animals at 2, 4, 6, 8, 12, and 24 hours ocrolflreocmtedthefrmoamrgeiintahlereaerarveviina otfheall post-injection. At the time of wsnatesocrreoodbptsaaytin(eradopopmfrrotoxenimmpatethreealtypuors4et8e-rhuinotouirrlsvecpneoansttr-ciiafvnuajgeecodtfioaenna)dc,hseaanppiapmrraaoltx.eidnaAitlnelt'loyssa2e0mrpulmmLesaonwfgerbelood mcaeilnltualianr afrtaectmipoenrsa.tureThoefbl-o20odCsa1m0plCesunwteirle sthhiepnpesdtotroedthien Sapofrneseozrer sop to Pathology tpAhotestt-peiornsmjtieencratitioioronn)v,eonfaantcihamevaale,sxpweaenrdriemeenxatsnaaelnsgtuhpiehntaaistzeeedd(.awpiptArhnoxaisbmabadrtieeuvlniyapte4en8dt-ohbgoarurorbssistanle,crobplseyd via pIelxvaaecmrei,dnatobniiloend,rywaasnidcen,boottthhceonnkdiudfcnrteoeyzdse,n fbrhyoonwpelveeaarcc,ihngatniiisnnsaulaeswferwreeeerzeecroclollseleteccttetedod.maanidnTthmaeeidnwihaaoitsely dtiesmcpaerrdaetdu.re of -20C +10C. After tissue/bile collection, the animals were Shipomf Teisnsutes EKAifEdt.neery&sPco.wmCep.rl,eetiBsloednngt.off2ro-tzh3ee-n0i9n(,o-n1i93fd5eryBpuhisaches)eAvtteohneutebh,eloSSotpd.onssPaoamurpll,e(sJ,MaNm,elsi5v5eD1r.0s5,J)o.hbnisloeTn,h,a a3nMd cHSNopIlolnedscooteresdinsointreatshcpicosenpstsitbuladenyy fnroorersptahorenesirtbehitelesinettyiorenfsouralntdtshediprsaepnsoaeslniyttseiidosn oinofftthhtiehse srsaeampmpoperltse.s. Statistical Analyses No statistical analyses were required by the protocol. cation of Raw Data. Record: Final Rey rTehteairnaewddaitna,therecaorrcdhsi,vesandofaHnHIoriigninaaclcorsdiagnnceed wciotphy HoNfI tShOeP.Final report will be 606817 Page 11 of 24 RESULTS BodyWeights Individual body weights at initiation are in Table 1. HHI 6329-134 Clinical Observations tIhnrdoiuvgihduoault cthleinisctauldy.signs are in Table 2. All five animals appeared normal Pathology Aelxlamianneidmalgsrosssulryvivwheedntosactreirfmiicneadt.ion of the experimental phase and were not DISCUSSION Trhaebbiltesvelwheonf 20, 200, and asydsmtienmiisctereexdposasurea 1,000 mg/kg. All sofinTgl-e605i2ntrwaasveneovuasluaitnejdectiinonfematalelevaellbsinoof 0, animals appeared normal throughout the study 2, following administration of this material. SIGNATURE SSttuedvyenDHi.recGtloarza UN Acute Toxicology p--bat--e 2AAS REFERENCE 1. NIH Publication No. 86-23 (revised 1985). 6003818 Page 12 of 24 Individual TaBboldey 1 Weights (g) HUI 6329-134 LeDvoseel Animal Grow (ma/ke) Sex Number Day0 1 0 Female Fs2548 3,031 . 2 2 Female F52549 2,921 3 2 Female F52559 2,813 4 200 Female Fs2566 2,912 5 1,000 Female F52567 2,853 600819 Grow 1 Dose Level (me/ka) 0 Page 13 of 24 Table 2 Individual Clinical Signs HHI 6329-134 Animal Hour Sex= Number _Observation~~ O05 2. & 78 4 Female F52548 Appeared normal So vy 2 2 Female F52549 Appeared normal Sod ry 3 20 Female F52559 Appeared normal Soy 4 200 Female F52566 Appeared normal Soy 5 1,000 Female F52567 Appeared normal SoS 7 Indicates condition exists. 606820 Page 14 of 24 APPENDIX A Protocol TP8084.PK HWI 6329-134 000821 @ MPWOASITbSOsFTFIwCOiEtsN]Ro0Sax I7183N4128 Page 15 of 24 + CORNING nur, Sponsor: St. Paul,mMinnesota PROTOCOL TPg0s4. PK Study Title: Single-Dose IOnftrTa-v6e0n5o2usinPhRaarbmbaictoskinetic study | Date: November 9, 1994 RBerforningLaboratory: MHa3ad3zi0ls1eotnKo,ninWsWimisascncoonnBssoiiunnl,ev5aI3rn7dc0.4 LaboprorjectaIdetntioficratiyon: HHI 6329-134 Nene an ni EB wo 000822 Page 16 of 24 STUDY IDENTIFICATION Single-Dose IonftrTa-v6e0n5o2usinPhRaarbmbaictoskinetic Study HHI No. Test Material Sponsor Sponsor's Representative Study Director Study Location PrEoxppoesreidmeSnttuadly STtiamrettaDbaltee DErxapfetriRmeenptoarlt DTaetremination Date 6329-134 T-6052 T3oMxicology Services S2t2.0-P2aEu-l0,2 M3NM C5e5n1t4e4r John L. Butenhoff, ph 2T2o0x-i2cEo-l0o2gy3MSeCrevnitceers S(t6.12)Pau7l3,3-1M9N62 55144 HSatzelveetnonM.WiGslcaoznasin, P.0. Box 7545 Inc. M(a6d08i)son2,41-WI729253707-7545 BHuaizlldeitnogn NWoi.sc3onsin, Inc. M3a8d0i2soPna,ckWeIrs 5A3v7e0n4ue WWeeeekk ooff NNoovveemmbbeerr 77,, 11999944 Week of December 12, 1994 PPa8ge082 4P.K. - 000823 Page 17 of 24 1. study PPa8ge0834p.K Single-Dose Intravenous Pharmacokinetic Study fn Rabbits 2. apTduanrniipnaoiisseetsesretdheasleavelsinogflesyistnetmriacveneoxupsosuirnjeecwthieonnthe test materia) is 3. RegulatoryCompliance to repeip: ee[xahncioesrpatstitoourndy,ythPwairtlalcatnibaceleycsRoinesdguocuftletdahteinitoeasnctcsom/radStaetnrcaieGnaludwiiadmteirh]xdittnoshene,sssfwoiltlsowiinngg Good cCoonndduccrteeadc:ions solubility, homogeneity, and stability witl not be [[X1] 2C1onCdFuRct58as(FaDAN)onregulated Study [[']] 4400 CCFFRR 719620 ((EEPPAA--TFSICFAR)A) [[[1115Nc9o(t8iN1fo)ih3cs0aatn(iFoMinon.aNlo)3.85(3001f3c(0J)a(pJaanpeasneeseWaHFOFR)M) - ) fYwaeotlrfgaptrhoreroscAecdttuhreReesgsutilunadtytihodinosse.sprnootItoncoutlnheneacorepesisniainroincloyomfpltihaenceSpwoirtehnrt1hedsAntiumdayl work. duplicate. sry neestocl 4. QutC(hhaWeelI)iJbtuuySaotlrAaoisnctsdoyular,rAdassnsutcrueadnyceconUdniutct,InaancdcotrhdeanFcienalwitrhepHoarztlewitloln. Wbeesaeuedciitcedby Operating Procedures (SOPs) and pol jens' S. TestMaterial A. TI-d6e0n5t2ification B. Ph(yTosibcealdoDceusmcernitpetdioinn the raw data) C. bgdPoeusttteeu grsmnpaioar tnnneasdrtoiii rSaotlnaa/sst bvsieu(lhmiy iiencstcllyeurdeisnpgonusnidbeirlitteystfocronpduirtiitoynsa)n,d s"tSabaim]plietyof tHhaaokkmeeonng))enwewriialttlyi,obneansdIefnstrteatqbouielmstiihttxeeytduSrapebno(yasnl)styohsreefsoSrpwoicnloslnocrbe.entrtaTathkieeosnne,bse`sfsmooptrluessi1s(riyf, termination for possible analysis. afterexper imectos 606324 Page 18 of 24 D. RSotoomragteemperature PPa8ge084 4P.K E. ReRseseerrvveeSsaammpplleess will not be required for this study. F. ftRnheyetenufintnuiasolendretpeosrtt,matuenrlieasls dwiirllectbeeddiostchearrwdiesdeabfytetrheisSspuaanncceerof G. SAasferteyqPurireecdaubtyioHnMIs SOPs and policies 6. ControlMaterial A. SItdeernitliefiacatteironfor injection B. PhClyesairc,alcDoelosrclreispstiToiqnuid - ; C. cTPohnesuipduerarrinetddySittaonadbtbiesltiyaatdbyeiqluiattyeofforthithseUpSuPrpgorsaedse omaftetrhiiasl stiusdy. D. SRteofrriaggeerated E. ReSeseerSveectSiaomnp,le5.s E. Reserve Samples : F. RaAenntdyewnritelimloainnnoitngbecodnitrsoclardmeadteraifatlermaiyssubeancueseodfftohreoFtihnearl treesptoirnsg. G. SAasferteyqPuirreecdaubtyioHNnIs SOPs and policies 7. ExperimentalDestan A. Animals 0 1) fSooeectses (2) Srtar:ai(nN/ZSHo)SuPrFc/eHRP, Inc. (3) AAdgueltat Initiation 600825 Page 19 of 24 (4) 2M50e3a.it5Iankigthiattion P1a0g8e0584P.K (5) N5umfbeemralaensdSex (6) IIdnedinvtiidfuiaclatniuonnbered ear tag (7) Husbandry (2) Hfonudsiivindgually, (heavy gauge} in screen-botton stainless steel cages (b) tAF{hoPeHmoIedmaaFsenueurdefsda,ctaumIroncue.nr)t.foorfTnhLueatbrofiortaaidtoonr1ay5l rRoaubtbiintelDyietanHaFnys#e5d32b6y environmental contaminants. componente any - . (c) hJfpMoaadarrttdeeInsrrieebslaisert,ceutneaadnnfdarloySnzpeeadcnifbaiyuetdWoWmmIaitcifrcoorbsityooslttoaeglmi.cdailsSsacomolpnvlteeedsntsoofianticehee, organophosphateelse,nenantds', chhleoarviynmaetteadlsh,ydrocarbons. @4) Cftohnatteawmoniunladsntsinterferecownittahmintahinstsstiundyt.he food or water (e)) EsEentvimtroonammaeinnnttainonatrtoetmsperfaotrurteheofani1m9alC relative hunidity of 50% rtooo2m3:wi.ll 4 be Tight/12-hour dark cycle.420%, and a 12-haur 0f) fAcecliinnationdays (8) SnaBodaeseqaeundialmtaaenlenhoueifnnacTbloetebrhtsvtioaiofnAudnseoilxbmytoardnaplyosoawrneiihmgeahlatsltahwciclolrdbiengputrochHaUsIedSOPvso.thaAyn is placed on esi (9) bJtHahicesuktgoarrnsoiinucanatlldlyoii,fnfffco"otrhhrmoeiSaitcpNieeecowcnbiaeZeocesnaatuTSsateenhildiesocofWtshipitneothceeniealsal.rbgienoamroaubnbtitorhas been 0006826 Page 20 of 24 8. DoAsdeainistration (1) TestGroups P8084. PK Page 6 Group Dnoas/ekLae)v*el NFuemmbaelresof 1 2 0 (Control) 1 2 3 20 1 1 45 1200000 11 a GTrhoeupdsose1-4voalnudmeawpiplrloxibmeat0e.l5yml1/.0kgmlf/okrg of body weight (depending on the test material) for Group bulk 5. density of the C. DosingProcedures 1) DosingRoute Intravenous injection into a marginal approximately 30 to 60 seconds. ear vein over : (2) sRysteenifacorexsDpoossoiurnegn.Rou.te Intravenous injection is an acceptable route to assess (3) DosingDuration Single dose (4) Dose Preparation The test material will injection to achieve a be diluted with sterile specific concentration water for dose level in Groups 1-4. The test material for will each be administered undiluted using the bulk density at to the 1,000 determine mg/kg dose level, the dose volume. bIonddyivwiediuaglht dotsaeksenwjilulstbebefcoarlecutleasttemdatbeasreidalon the animal's administration. The prepared test mixtures at room temperature until administration. will be stored 0. O(1)bsCleinricvao)afAOtbnsiiemroavlanstions The animals after treatnent. toxicity at will be observed for clinical signs of approximately 0.5, 2.0, 4.0, 24, and 48 hours 600827 Page 21 of 24 2 i PTaPg8e0784P.K (3) sampleCollections a) fx requencySEN hourspost- inject on (b) aNum ofb Aniemar ls (c) 4gfBM u,rlroo6imo,e cdue8lois,at tafrhmC1ep2raloh ,erlstelaeaeo nrr(cdyatvp2id `ipo4aorrnohftxorhiuoemrnasctatethlehyemt4aerrngiLi)znaatwliiolln`saobrfevecSrooenllaetct2e,d pdAoopscpturenorexiniotmreadtveelinnyat2ch0aevaalraowoffdeabatlcaoh)opdoawsin(litaln-catilbupaeelatcobtvttoihateeiunneseditmoproobtse the s2asan0emicpaBrldLoelpssosfywdiub(lrl4lio8nogbdheotwushrietslorspetobdusedtya-c,tionlrj1lo1eeo5ccm0tt',ieodn1)f.rpoonAsppepiomriosesiincrtheoy i ctcfeeerlnmaltpcuretliriafaorutnguserdes,toofraendd-20tinhCea+s1ef0praeCre.azteerThsseeettresunetnppohararmadaattiuesrrdsoe)alsiaeonarda2tmHpaennd after expefrriamcetnitoanls wtielrlminbaetisoenn.t frozen to. the Smoumer Samples will be shipped to: BWJladnEge.sE2D-.3E6JP.o6hC9nson S9t3.5 PBauuslh,ABvNenu5e5106 J(a61n2e)s 07.78-J5o2h3n4sonprwioirl tobethneotisfhiipemdenbty otfeltehpehonseenmaetres. E. Termination m UrgmAneronscorycosihrsbaedundennidudne.laclercoodApndSndsyaiiyicmtnraigeolixnsfad,imuicirnnwieiansltagalinomtnodbhreeDiaebnssaudutntbudhj3d1esy1ctoeardbsnatoocrriamyfaitc<jeiaedtrroionmntasedue Caanveas,theatndizeedxswanigtuhinsaotdeidu.m pentobarbciotnadli,tioblnedwitvsiabhe vena 0G828 Page 22 of 24 TPB084.PK. Page 8 (2) ScheduledSacrifice At approximately 48 hours post-injection, animals pseunrtvoibvairnbgittaol,tebrlmeidnatviiaonthweillvenbae caanveas,theatnidzeedxswaintghuisnoadtieudm An abbreviated done, however, gross necropsy examination tissues will be collected. will not be (2) SamleCollection dTuhreinwgholtehe Tsitvuedry,andsabcirilfeicferdomineaachmoarniibmualnddycionngdition, or surviving kidneys from to termination will be each animal will also collected. be collected. Both The ctoilslseucestiownillandbeptlheancepdlacoend driny ice immediately after a freezer set to maintain a temperature of -20C 110C. samples. fThreozetnisosnuesdry(liicveer,tobithlee!, Spkoidnnseoyrs)afwtielrl exbpeersiemnetntal termination. person listed iTnheSescatmipolnes7w.i0l.l(3)b.e(cs)h.ippeThde toSpotnhseor is responsible for the retention and disposition of the ! F. StWoatsitsattiisctailcAalnalaynsaelsyses are required. 8. Report A final report including those items Tisted below will be submitted. Description of the test and control materials Description of the test system Procedures Dates of experimental initiation Description of any toxic effects and termination Gross pathology findings/gross pathology report (ifapplicable) 000329 Page 23 of 24 PPa8ge0834P.K 9. LaofOcraccicaegitpilitnioaatnlnactoeedfatoRaafua,dwtiDhtoeirantgFcaio.ntpaRhileeescrsoettrpuhdoedrsryt,e.oadfun,rdiWhnwFeiginlnliatt]hsbReeperfapiovongaarrileltsasbrelpeaonrdattbeHfMoIreto raelltaionreidginianlthpeapaerrchdiatvae,s oifncHlWuIdinagccotrhdoisengittoemH1I1sStOePd. belioswcwoimpp)letreed, DPorsoetocporlepaarnadtipornotorceoclordasmendments InB-o1diyfeweriegchotrsds ObDsoeservaadtmifonnisstration PSaamtphloelogcyolRleeccotridosn records FSitnuadly rceoprorretspo(nodriegnicneal signed copy) nTohte bfeolalrocwhiinvgedsuwpiptohrtithneg srteucdoyrdsdatwai.ll be retained at HNI but will WAnaitmearlanraelcyesiips't/raecccolridnsation records IRAnneisfntrarilugmeerrnaottoomrcatleainmbdprearftariteoeunzreeranadtnedmmaphieunrmtaietdnuiratenycerreercceoocrroddrssds ) 600830 Page 24 of 24 TPPa8g0e841.0PK PROTOCOL APPROVAL Denn 2. Huttay SJpoohnnsoLr. 'sBuRteepnrheosfe?n,taPthiDve MH SSttuedvyenDiH.recGltaozra , HAaczulteetoTnoxWiicsocloongsyin, Inc. 5 QualsietyntAastsiuvreance Unit Hazleton Wisconsin, Inc. (6329-134. protdskl) I t5-5y bate Date aqu "/aloy Date J . 00331 9.12 Analytical protocol AMDT-111694.1 0006832 3M Environmental Laboratory Protocol - Analytical Study Single-Dose Intravenous Pharmacokinetic Study of T-6052 in Rabbits In-Vivo Study Reference Number: HWI#6329-134 Study Number: AMDT-111694.1 Test Substance: FC-120 (T-6052) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facility: 3M Environmental Technology and Services 935 Bush Avenue St. Paul, MN 55106 Proposed Initiation Date: July 25, 1995 Proposed Completion Date: August 25, 1995 Method Numbers and Revisions: AMDT-M-1-0, Thermal Extraction of Fluoride by means ofa Modified Dohrmann DX2000 Organic Halide Analyzer-Liver AMDT-M-2-0, Fluoride Measurement by Means of an Orion EA940 Expandable Ton Analyzer AMDT-M-4-0, AMDT-M-5-0, AMDT-M-8-0, Extraction of Fluorochemicals from Rabbit Liver AElneacltyrsoissproafyRMababsist SLpievcetrrEoxmtertarcyt for Fluorochemicals Using AnalysisofFluoride Using the Skalar Segmented Flow Analyzer with lon Selective Electrode AMDT-M-14-0, Thermal Extraction of Fluoride by Means ofa Modified Dohrmann DX2000 Organic Halide Analyzer-Serum Author: James D. Johnson Approved By: L es D. JgBfison tudy Dirdetor foo) ps Date Plo 1. BueBally 1of5ots~ John Butenhoff, PhD Date Sponsor Representative 0006833 PURPOSE oThfaisssutbusdeyquisenpterdfeorrmmaeldaibnsoorrpdteirotnosptruodvyid(eHWpIh#a6r3m2a9co-k1i3n5e)t.ic data for the assessment 2.0 TEST MATERIALS 2.1 Test, Control, and 2.11 Analytical Reference Reference Substances Substance: and Matrices FC-95, lot 161 or 171. They are equivalent. 22..11.32 AAnnaallyyttiiccaall CRoenfterroelncSeuMbasttrainxc:e:BNovoinnee liver and bovine serum 2.1.4 Analytical Control Matrix: Bovine liver and bovine serum 2li.v2erS),ouSricgemaofCMhaetmeirciaallsC:o3mMpaInCyP(/2P.C1.P2,Di2v.i1s.i4o-nse(r2u.m1).1), grocery store (2.12, 2.1.4- a2.n3d N1ucmonbterorloafniTmeaslt.OatnhderCboinotlrooglicSaalmptlisessu:esL(ikviedrneayn,dbsileer,ucmelflruloamr 4frtaecsttioann)imwialllsbe available for analysisif deemed appropriate by the Study Director. 2.4 the HIdeWntiafniicmaatlioindeonftiTfeisctataionndnCuomnbterrolwhSiacmhplceonss:isTthseosafamlpeltetesr are identified and five digit using number, plus the tissue identity, and day identity (serum). 25 Purity and Strength of Reference Substance: To be determined by Sponsor. 2.6 Stability of Reference Substance: To be determined by Sponsor. 2.7 -20:+ Storage 10C (C2.o1n.d2,it2i.o1.n4s).foTreTstesatndMaCtoenrtiraolls:saRmopolems temperature (2.1.1), will be received according to AMDT-5-10-0. G2.L8PDiRsepgousliattiioonnofforSptehecitmiemnesp:erBiioodlorgeiqcuailretdifssouresstaunddiefsluliodnsgweirllthbaen r2e8tadianyesd.peTrhis study is in retained. parallel with a 28 day dermal absorption study so all tissues will be 2.9 Safety Precautions: Refer to appropriate MSDS. Wear appropriate laboratory attire. Use caution when handling knivesfor cutting the samples. 000334 2 3.0 EXPERIMENTAL - Overview aTannhaaellyyttsiiiscssaulfeostrefsftrlsouocmraiannnebimecaoplmesprodfouorsnmededsd..asATdtehsetchrseicdbrieesdecnr(ieHntWgiIofn#oo6rf3tf2lh9u-eo1ri3Sd4te)ud,inyalDriievreaervcatvioilraa,bcaloemsbefruoisretsioofn c(folSuneoferiiMrnemetthihnoedtsph-re-elnsieevxnerct.eSoeEclftesicpotenrc)oisfipiscrtahmyeolmaeapcspusrloespsrp.ieactTterhoeamnemaatltryesyriiwsailltlobpbereienspgeensrttfuodderifmeiedndiitsiinvaeorddatehr tfour wpbiielorltfrlbauenosrpfoeordsreimcseatdne.enstuIlifnftothnheiecmlaaitvceeirrd.iaslaAlnitas(lsyAismmiimsloaonrfituloimpv)eer.rffTlohuroitrsootmoaacltteoarrnigeaaslnuilicfsonfnolitucoetaxicnpieedcwatinelildont,o be it provide information as to the extentofpersistence. 4.0 EXPERIMENTA-LMethods 4.1 Liver and Serum screening methods: (attached) D4.o1h.1rmAaMnDnTD-XM2-010-00,OrTghaenrimcaHlalExitdreacAtniaolnyzoefrF-lLuiovreirde by Means ofa Modified 4E.x1p.a2ndAaMbDlTe-oMn-2A-n0a,lyFzleuroride Measurement by Means of an Orion EA940 4.1.3 AMDT-M-4-0, ExtractionofFluorochemicals from Rabbit Liver 4Us.i1n4gAEMleDcTt-roMs-p5ra-y0M,aAsnsalSypseicstroofmReatbrbyit Liver Extract for Fluorochemicals 4A.n1a.l5yAzeMrDwTi-tMh-J8o-n0S,eleAcntailvyesiEsleocftrFoldueoride Using the Skalar Segmented Flow 4M.o1d.i6fiAeMdDDTo-hMr-m1a4n-n0,DXTh2e0r0ma0l OrEgxatnriacctiHoanliodfe Fluoride by Means Analyzer-Serum of a 06068353 5.0 DATA ANALYSIS 5f.l1uoDriadteapeRreptoirsstuienogr: flDuaitd,a owrilalsbFeCr-e1p2o0rt(eedleacstraocsopnrcayenmtarsatsiosnpe(cwteriogmhett/rwye)igphotr)unoift oifntcilsudseueavoerrfalgueids.anStdatsitsatnidcsarudsedde,viaatttihoensdifsrcormetdiiofnfeorfentthedoSsteudgyroDuirpesctIofr,nemceasysary, simple statistical tests statistical difference. such as the Student's t test may be applied to determine 6.0 MAINTENANCE OF RAW DATA AND RECORDS 6.1 Raw approved Data final raepnodrtR,eacnodrdesl:ectRroanwic data, data awiplplrobevemdaipnrottaoicnoeld, ianpptrhoepAriMatDe Tspaerccihmievness., EFER] 7.1 AMDT-S-10-0, Sample Tracking System 8D.o1hArMmaDnTn-MD-X12-000,0 Thermal Organic HEaxltriadcetAinonaloyfzeFrl-uoLriivdeer by Means of a Modified 8E.x2paAnMdDaTbl-eM-Jo2n-A0n,alFylzueorride Measurement by Means of an Orion EA940 8.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver 8.4 AMDT-M-5-0, Analysis of Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry A8.n5alAyzMeDrTw-iMt-h8I-on0,SeAlencatliyvseisEloefctFrloudoeride Using the Skalar Segmented Flow 8D.o6hrAmMaDnTn-MD-X1240-000,TOhregramniaclHEaxltirdaectAinoanloyfzeFrl-uoSreirduemby Means ofa Modified 000336, 3M Environmental Laboratory Method `Thermal Extraction of Fluoride by Means ofa Modified Dohrmann DX2000 Organic Halide Analyzer - Liver Method Identification Number: AMDT-M-1 Revision Number: 0 Adoption Date: /o- f-1 Revision Date: None _-- Author: Rich Youngblom Approved by: Zoup Lead ---- /of3/7s `Date Quality AsLsurVaknocPe E L2-2- 1.5 Date Software: MS Word 5.1a Affected Documents: AMDTE-xMp-an2daFbllueorliodneAMneaalsyuzrerement by Means ofan Orion EA940 AMDT-OrEgPa-n3ic RHoaultiideneAnMaaliynzteernance ofaModified Dohrmann DX2000 006337 L0 SCOPE ,APPLICABLECOMPOUNDS, AND MATRICES fLl.1uoSricdoepef:roTmhivsarmieoutshomadtriiscfeosr.thTehoepefrlautoiriodneoisfatyDpiocharllmyacnonllDecXt2e0d0in0TwIhSeAnBitsiosluutsieodn tfoorexatnraalcytsis `with an ion selective electrode. 11..23 MAaptprliicceasb:leBiCoolomgpiocaulntdisss:ueFs,lupoarrtoicchuelmairlcyallisveor.r other fluorinated compounds. '2.D1X2F0l0uo0r,idleu,orfloucohreimnei,caelxtsr.action, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, 3.0 PRECAUTIONS 3.1 Glassware and exhaust gases can be extremely hot, 3.2 3.3 PGrleassssuwrairzeedisgafsreagsi,lep,robpreorkecnomgplarsesssmeadygcaasuhsaenidnljiurnigesp.ractices required. 33..54 PSootlevnetnitalbbaisoehdazsaarmdpsledsuematoythfelabshi,olmoagiycanlemeadtrtioceasl.loUwtsheeamptproodprriyadteopwenrsobneaflorperosttaercttiinvgerun. equipment. 4.0 SUPPLIES AND MATERIALS 4.1 Compressed Oxygen, Hydrocarbon free, regulated to 30 PSL 4.2 Compressed Helium, High Purity Grade, regulated to 45 PSL. 4.3 4.4 Quartz Quartz glass glass sample boat with combustion tube, TeflonTM Reliance Gtluabsisng,G-D9o4h0r5m-a0n1n2 8or90e-qu0i9v7aloern`te,quivalent. 44..65 SOarimopnle94c0ol9l9e9ctTiootnavliaIlosn,iHcDSPtrEe.ngth Adjustment Buffer (TISAB IT ) or equivalent, 4.7 Milli-QTM water 44..98 PAocltyisvtatyerdenCehapirpceotatle,s.E. Merck 2005 or equivalent. 44..1110 MHiasmcielltloanneSoyursinlgaeboorrateqouriyvgallaensts.ware 5.1 5.2 RIoBsMemcoomupnattiDbolher3m8a6nnorD4X8260c0o0mpOurtgearn.ic Halide Analyzer, modified for fluoride extraction. 5.3 DX2000 software, version 1.00, modified for fluoride extraction. 5.4 Excel Spreadsheet, version 5.0 or greater 6.0 INTERFERENCES EE -- 6.1 Sample size is limited to approximately 150 mg, depending on sample moisture content. This may vary from matrix to matrix. . 2 C0G838 2.0 SAMPLE HANDLING s7a.1mpSlaem.plUesseafroerncoetpstoorbeprhoabnedlteodtrwaintshfebratriesshuaensd,s. Fluoride may leach from the skin to the `c7ml:ee2tahSnaansmocplal,lpeoesrol mfaenltdihvcaeunrtoatlri-enwgacbutotearrfdsr.oolmuTtfhireoonzceaunftttleiirnvgeerabcaohanrdldipvalenraidcsescdcuatiln,paeltasrheoduladndbelacbleelaendewdewiigthh bwoaatte.r,Use a 8.1 PreparationofCalibration Standards R8e.f1e1rTthoeprsottaoncdoalrdfsorrtehqautiprreodjefcotr.each project will need to be appropriate for that individual project. 88..11.32 FTyoprircaablbliyt 5li0v-e5r0s0tupdipems,FuCs-eb9e5eifnlmievterhaasnotlhesmtaatnrdiaxr.dsCaurte auspeide.ceoffrozenbeefliver (100 150 mg) and weigh it in a labeled and tared weigh boat. 82 Calibration - Overview cTuhrevenoirsmraelquciarleidbrtahteipornoicsetdhuerfeluliosrtieddebceulrovwe i(sAuMsDedT.-M-2). However, ifan optional spiked liver 88a..n22d..21coTAympbicucasaltliilbynr,agttithohrneecemururesvpielnifcgoarttehtsehoefseaDmaXec2hm0e0stt0haonisddasgreadnneadrnadmtaeftdirvibexycotsnypcpieeknaitsnrgatthsieaomnsspaolmefpslsewtsiatnthdoakbrednsotwewsintlelsd.tbea:ndards. spiked. R8.e2c.o3veSrteadndFar(dugc)uornvethweilyl-abxeisp.loGtteenderaastMeaasrsegSrpeiskseidonFc(uurgv)eoanndthcealxc-ualxaitseatnhed eSqtuaantdiaorndfMoarsthse line 8a.n2d.4thMea1s2svaSlpuiek.edF (ug) = (Amount spiked in mL) x ( Conc. ofstandard in ppm)x (0.6004) 8.2.5 Standard*MFaCs-s95Reicso6v0e.r0e4d%FF(tuhge)re=fo(rTeI0S.A6B00v4oilsutmhee finacmtLor)usxe(dOrtoiocnonrveeardtinFgC-in95pptmo )F 8.3 Calibration - Procedure 8b83e3..u11s.e1dSitRfautrhtneU2pporrevmiooruse sCalmepalneCsyccolnetsaiwnheednhisgtahrtcionngceinntsrtartuimoennstoefafclhuodraiyd.e.More clean cycles may 88.33..22.1BlParnepkasre sample using the same methods and typeofmatrix as the test sample. (8130.02.215F0ormrga)bbfiotrsbtuldainekss,.usebeefliver as the matrix. Prepare at least 3 samplesofbeefliver a8n.3a.l2y.z3e sPaumtpslaempalcceoirndiDnoghtrommaentnhobodatA.MDCoTm-bMu-s2t efaocrhthseafmopnlesealsecdteisvcereilbeecdtriondseeacntailoynsi9s.,0 and 3 000839 8.3.2.4 Forrabbit before proceeding studies, with the the meter reading calibration, Bum foar blank samples sample should be 0.03 ppm or lower judgementof hours). the operator the reading is stable with until respect tthoishilsitmoirticiaslrreeaacdhiendg,so(rpurnetviiloiunst4h8e 83.2.5 For non-cabbit before proceeding with studies, the blank the calibration. readings should reach a predetermined ion concentration c8.o3m.b2u.s6tiIotnm.ay be necessary to mix approximately 50 mgofcharcoal with the sample to aid 8.3.3 Standard Curve 8.3.3.1 labeled Weigh out at weigh boats. lFeoarstra1b5bimtatsrtuidxiessa,mpwleeisgh(51s0t0a-n1d5a0rdmsgwbieetf3hlirevpelricsaatmepsleesa.ch)Reincotarrdewdeiagnd 88s33t.u3d3.y.32daStPtaa.lratcSetwoiwrteehittghhheeemldaobtwereeisfxtlsisavtamenrpdlsaerasdmopcnloendcrieynntDircoaehtoirromnia.cneUnpsasciakmnsapglteoHakbmeoiaerlp.ttohnesmyrfirnogzee,neujnetcitlausfeidx.edhts in oQruainnttihteyboeftehfleivsetra.ndard on or in the matrix. For rabbit studies, use 4 uL ofstandard and eject iton `8m3a.y3b4e uAsteldeaatsttherdeipslcirceattieosnsohfotuhled banealuyssetd.forthe lowest standard concentration; more replicates or88le.3pd3.l.ri33ec..ap65tleisRcC,auotrnmeubanaulnalsdnt1ot5thtahhsteteratsnhsdaeaamnrmpdpelslwe.earfIesofprdloietncsheacattrreieswbptielaldinlcdiaabntreeds.ueisscetIsindiodgtninoci9afc.tia3celacianunntllddayatatednaitafhtlfheeyarzrteeentgtahrceefcsnrosoeirmowdnitrnhcegepulotiotchaAetMre DtrewTpo-lMac-e2s ,the l8e.a3s.t3.07.95W,hceonnsaullltswtiatndhasrudpserhvaivseorb.een run, calculate the r2. r2 must be at least e. 0.95. Ifit is not at 8.3.3.8 A new changed. New standard standard curve curve should be run when the combustion tube or sample `may also be run at the discretion oftheanalyst. matrix is 8.4 Storage Conditions for Standards 8T8y.4p4..i21caSlNtleoyr,wasgFteCanr-de9aq5rudissrteaamnredeansrttdsofsroesrdhosauttlardnodboaermdptsreeamrppeaerdreeadtpueartnedleeiannsttplooannstctiehce2sicmnrodeniwtvihtd.oupalbosrttalnedsards used, 9.0 PROCEDURES 9.1 Typical Operating Conditions: 9.1.1 9.1.2 OCoxmybguesntainodn Hteubleiutemmfpleroawt=ur5e0=cc9/5m0inCu.te. 99..11..43 BVaapkoeritziamtei=on3/0D0rysiencgontdism.e = 240 seconds. 99..22.1SItfarttheUpprPorgorcaemduisrneo:t started, start the EOX program on the PC, 9.2.2 Open the SYSTEM SETUP window. 99..22..34 PCultostehethfeumSaYceSTmoEdMulSeEaTnUdPthweicnedllowi.n the READYmode. 4 006840 5:25 found Wjhn etnhe tCheEoLvLenChHaEs CreKacmheendut.he READY temperature, run the CLEAN BOATprogram 92.6 See AMDT-EP-3 for detailsofthe Dohrmann software. m99i.33.x1SaapOmpppreolnxeitmEhaxetteSrlaAycMt5iP0oLnmEgPorHfocAcheTadrCucrHoea:alndwiptlhactehethseamspalmeplteo aiindtcheomBbOusAtTi. It may be necessary to 9s.ho3u2ldCallossoebSe AmiMxPeLdEin HwhAiTleCHes.tablishing the baseline and when gencroant.inIgF yi the s1tduonne,cucrhvaer,coal $la:b3e.l3eddsdampalppercooplrliescteivonolviuaml.e oTyfpTiIcaSlAlBy 0s.o6lumtLiontoo 2.0 mL of 1:1 TISAB:Milli-QTM water mixture, 151m:1LTaIrSeAuBse:dM.ilFlo-rQrTMawba.tesrtumdiixest,urueseto1a.0 or i99n.3c3.h.e45s.RPluGanacsetehtseheErevOliXeala-soSedOtdLhuIartDitnShgepptryiorpooglfryastmihsefmoCuuOsntdMBbinuUbtSbhleTeRItOUhrNNouTmgUehnBtuhE.eiTsIiSnAtDh,e TISAB at least 0.25 t95o33..th75e TIWfIhuSnedAniBltuhtteoeEmdaOTkIXeSpA1r:Bo1gwTraIasSmAuiBss:efMdiintlioslchieo-dl,iQeTMrc,etmthoevseatmhpeleco,llaedcdtiaonn evqiaulalfrvoomlutmheeOcfomMbiulsltii-onQTMtuwbae,ter r2:e3m.o8vReinasneytThIeSeAndBorfetmhaeincionmgbuosnttihoenttuubbe.e with Mill-QMTM water and wipe with a KIMWIPE to b5voi2tt.ht9oamOcwpoitettnohntahisepppsaeatumdlpaalpeoprlhioacttahctehorrasonirdmivrlaeacrmudouevvmeiecaden.oyutAr.edmIratoimpnaioynfgMbiaeslnhleifcer-soQsmTMatrwhyaettboeorsaetmr.sapAypsabhretciaclnesbeofrepmsoved 9b.o3a.t1t0o aCildosine tthheeChaltecahn, Cycle. added so ie 9.3.11 93.12 SRuumnplteheisCrLeEadAyNfoBr OanAalTyspirsobgyraimo.n selective electrode (AMDT-M-2), 9.4 Sample Calculations 9.4.1 Use the standard curve to calculate the sample value, 9.4.2 Sample Mass Recovered F (ug) = (TISAB vol in mL) x(Orion readin(gSilonppep) m -intercept) 10.0 VALIDATION a21n00a..l11y.s1iQsDuaaiilslisytayrSttCeaodrntbtrUyopalnCahlyezcikngSQamCplseasm:plOesn.ceTthheeQstCansdaamrdplceusrvareeistoesbteabtlhieshseadm,eeaacsh day of rdGeoopnnlcieecnaiintretaritisipnloointcasntpeeickuenesldseassrsaymt.hplefsirustsetdwtoorgepelniecraattese atrhee wsittahnidnar2d0c%urvoef.thEeascthancdoanrcdencturravtei,otnthhomenuJs5otwfebosotd mFFaltu2roirciPedrseeaNcniosdtiedobinoffoaeknrsedn2tA.c3r,cefuaernradecn3yc.:e PcrSoeemcepiosmuieontndhsao.ndddaecvceulroapcmyevnatriaensalwyhsiesnaanndalsyazmipnlgesaanmapliyessiosfindifferent 10.3 Other Validation Parameters: NA 5 GOG841 11.0 DATA ANALYSIS 11.1 Calculations 1LLI For the standard curve, use regression analysis in Excel, version 5.0 or greater. 11.1.2 To calculate the fluoride contraction in the sample, see method AMDT-M-2. 11.2 Analyzing the Data a1r1e2.wi1th2inm2u0s%t obfeeaatclheaostth0e.r9a5nodrtghreeaotuetrl.ie"rOuitslgireerast"ermtahyanb2e0e0x%colfudtehdeifatvewroaogfetohfethtohsreeetroeppolsi.cates less than 50%ofthe averageof those two. Any such outliers should be pointed `noted in the Final Report along with the reason it was considered an outlier. outin the dataand None 13.0 REFERENCES ~~~ = 1334.91, RreovsiesimoonuBn,tDDeochermmbaenrn1D9X932)000 Organic Halide Analyzer Operator's Manual (Manual 915- A1n3a.2lyzAeMrDT-M-2 Fluoride Measurement by Meansof an Orion EA940 Expandable Ion 13.3 AMDT-EP-3 Analyzer Routine Maintenance ofa Modified Dohrmann [DX2000 Organic Halide 14.0_REVISIONS Revision Number ReasonforChange Revision Dae 6 600842 3M Environmental Laboratory Method Fluoride Measurement by MeaAnnsaloyfzaenr Orion EA940 Expandable Ion Method Identification Number: AMDT-M-2 Revision Number: 0 --- Author: Rich Youngblom Approved By: Grod] Leader Adoption Date: /0-4-%5" Revision Date: None VEY 2S Date Quali7ty,ASsG uranL cB e AL Lo-#F1 Date Software: MS Word 5.1a ADfofhercmtaednnDoDcXu2m0e0n0tsO:rgaAnMiDcTH-alMi-d1e ATnahleyrzmearl Extractionof Fluoride by Means ofa Modified 1 6006343 10SCOPE,APPLICABLE COMPOUNDS.ANDMATRICES Hon Ton SCOPE: Analyzer. This method is for th calibration and operationof an Orion EA940 Expandable 1.2 APPLICABLE COMPOUNDS: Fluoride. P1H3OfA6P.P0L.ICABLE MATRICES: Liquid samples in an appropriate buffer solution, Preferred 2.0 KEYWORDS 2.1 Fluoride, fluorine, ion selective electrode 3.1 No hazards identified with this method. 444..-231 OOOrrriiiooonnn M994o40d09e90l979901T00ot00a0pl1poemnliefccltuSrotordrieednefgistltlhainnAgddasjroudlsutotrmioeennqut(iABvguaClfelfn)etr.oIr e(qTuIivSaAlBeInDt.) or equivalent, 44..54 MMaiglnleit-iQcTMswtiartbearrso.r equivalent. 44..76 SLaambptliesscuoelsl.ection vias, 44..98 PPolalsytsitcyr1e0n0empiLpetvtoelsu.metric flasks. 4.10 Miscellaneous laboratory glassware. 555.321 MOOarrgiinooenntMMioocddSeetlilr9EPl6Aa0t9e94.000 ESxolpiadndStaabtleeCIoomnbiAnnaatliyoznerFoluroerqiudievaelleenctt.rode or equivalent. $5$654 MOIirBcirMoonscRoofmStp2a3Etx2icbeilnlete53r.8f0a6(cooenrcla4yb3ln6eece(dooemnldpyuitfneeuresid(neogdnlOiyrfinuoesinend3geEdOrsifiouofstniw3anrEge)sO,orfitownar3eE).software). 6.0 INTERFERENCES &1 QM waisterrecwiolmlmgeennedreadlltyhabtrithneg psaHmpbleeattoorpnHeaorf6.06..0. A 1:1 mixture ofTISAB and sample/Mill- a6t:32rToShoaemmprtlaetemeptetehrmeaptseuarrmaeptluaresesthmaeraesytsateinfrdfraeecrdtdasftlwuseohrroieudlewdhmbeeeansctuohrneseimmseetnteten.rt wwIiattshsctrahleeicbrroaamttemedte.hnedsetdantdhaatrdtshewesraempsltierrbeed, 2 000842 u6n:4deArirelbeuctbrboldees. trapped under electrode can give emoncous readings. Make sure no air istrapped 7.1 No special handling necessary. 888..111..21PLMraeebpaeaslruarttheeioSfnOloamsfkLsCoa4l3fi0Tb.rI0a5St,Ai0Bo.n1I,lS0.ti5na,tnod15.a0r,1d0as0ndm5L pplapstmicF-v,olaulmoentgrwiicthfltashkes.date and your initials. 4lab2elePdipfleatstkes.0.05,0.1, 05, 1.0, and 1.5mLof 100 ppm fluoride standard into the appropriately 8.1.4 8.1.5 SAhdadkeaptphreoxfliamsaktsetloym3i0xmthLe osfolMuitilolnis.-QTM water to each flask. t8h1e 6flEaslkismionvaetreathbeuabirbbluebbslefsr.om the flasks by tipping the flasks ontheirsides and rolling theairin 88..11..87IBnrvienrgtatnhde vsohlaukmeethien ftlhaeskfslafsokrstuhpe tfointahlemi1x0i0nmg.Lmark with Milli-QTM water, 8.1.9 Record standards in Standards Log Book. 88..22.1CIaFlniebcreastsiaorny, remove tape from electrode filling hole. 888..222...432FEIijnlveectrthtaepefrleoewbcetdrrtooodpewseowtfitfthiolpflisilelnaigln.gsosloultuitoionnf.rom bottomofelectrode to wet lower seal, St8ho.e2m.em5catTniuhofena,cmteuutsreienrrga'snstdianntsdhtearruFcd-tsieowlniset.chtrcoodneceanretrtaytpiiocnalsolyfc0a.l0i5b,ra0t.e1d,b0y35d,ir1e.c0,t amnedas1u.r5epmpenmtFwritfhonlloowbilnagnk 8t8i..s22s..u76esRC.leecaonrdthteheelselcotpreodien bthyeraipnpsrionpgrwiiatthe Mloiglbloio-kQ."waterand wiping the sides down with ab 88.33.1SCtaolriabgraetCioonndsitatnidoanrsdsfoarreSsttaonrdedaradtrsoom temperature. 20PROCEDURES ~~ 000000 99.-111CaTlhiebrsaatmipolneatnodbeMemaesausurreemdennte,edSsttaonbdearmdixmeedtwhiotdh:TISAB recommended by the TISAB manufacturer. using the proportions 9e9.l.1e1.c.32trPAoldlaelcoieswaitnshsteeirrstbaedam,rplimenattkoheemssiuarxmepfltoheearraenfdaerwpelsnaeocceoaitnrhdesbusbbaebmflpoelrseetoirnnaspetprhetedinsugtnitdhpeelraettlehe.ecterloedcet.rodWeh.en the 91.4 same The rate assamtphelecaslhiobrualtdiobnestthaendsaardmse, temperature as the calibration standards and stirred at the 9-1:5 When the readings have stabilized, record the reading in the appropriate log book. 3 C0084: 9.2 Calibration And Measurement, Using Orion 3E Software: 9.2.1 Calibration: 9.2.1.1 Follow steps 8.2.1 t0 8.2.4. 992.2..11..32 TPhreesscoFmupnuctteironscKreeyen#w8il(lF8a)s.k you to confirm the numofbstaendarrds to be used, Mcoankceenatrnaytinoenceosfstahrey scthaanndagredss,toanthdewihneftohremratoironnoptraesbelnatnekd iastnodbcelicikncolnudCeOd NinTIthNeUcEal.ibration. s99t..a22b..i11l..i54zeOPdbl,asceperrevtsehsetAehleCerCcetEardoPidTnegisRnEotnAheDthfIieNrsgGtr.astpahnidcadridsoplnatyhoensttihre`pcloatmepuatnedr.cliWckhoennCtOheNrTeIaNdiUnEg.s have i99n.2t2.e.1r1.c.7e6pARtfeatpneedratctohrsertfeeilpnaat9li.o2sn.t.1a.n4Rdeaarcndod,rdt9.ht2eh.c1.os5mlfopopurett,ehirentwreirelclmeapdtii,nspialnnagdyscttoharenrdesallraodtpsie.oonfitnhtehecuarpvper,opariwaetleloagsbtohoek and click on CONTINUE. The calibration data is automatically copied to C:\Orion\Data\Calitxbt.. 992.2..22.1DaStelaecStprcetahdesrhNeeEt:W or OPEN from the FILE menu to open a new or existing spreadsheet 9t.o2s.t2o.r2eRdeactoaridn.the nameofthe spreadsheet used in the appropriate log book. 9.2.3 Fluoride Measurement: 9.2.3.1 9.23.2 EFnotlelrotwhestneapsme9.o2f.t1htehrsoaumgphle9.i2n.4the appropriate place on the screen. 9.2.3.3 Click on the NEW SAMPLE button a9.p2p.r3o.p4riWahteelnogthbeoroeka.dings have stabilized, click on the RECORD `button and write the resultin the 10.1 Quality Control: 102 Precision and Accuracy `1p0p.3m fOltuohreirdeVuaplitdoaatisoantuPraatreadmseotleurtisonAocfcofrludoirnigde.to Reference 13.2, the range ofdetectiions 0.02 11.1 Calculations None necessary. 11.2 Analyzing the Data None necessary. 2.0 ATTACHMENTS None 13.0 REFERENCES 4+ 000846 13.1 Orion Model EA940 Incorporated, 1991. Expandable fon Analyzer Instruction Manual, Orion Research R1e3.s2eaOrrcihonInMcoordpeolra9te6d0,901099S1o.lid State Combination Fluoride Electrode Instruction Manual, Orion 10 40REVISI0 ONS ~~ 0000 Revision Revision Number Reasonfor Change. Dae 5600847 3E M nvir Labo oratn orymental| Method Extraction of Fluorochemicals from Rabbit Livers SOP Identification Number: AMDT-M-4 Revision Number: 0 -_-- Adoption Date: /o0-7/-5+ Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: up Leader 44Bh Quality Assurance /Jo--3/-25 Date l3r15 Date MSAfoaffsetscwtaSerpdee:cDtoMrcouSsmceWonpotyrs.d:, M6-.05, Analysisof Rabbit Extract for Fluorochemicals Using Electrospray 000848 1 1.1 EStchoyple:acTehtaitsemies tuhsoedd tiosexfotrratchetfelxutorraoctcihoenmoifcfallusofrroocmhetmhieclailvesrsfrfoormarnaablbyistislibvyers, 11..32 Alpepcluiocgapbrlaey Cmoamssposupencdtsro:scopFyl:uorochemicals Matrices: Rabbit Livers. or other fluorinated compounds. 2.1 Fcloumoprooucnhdemsi,caexltsr,acrtaibobni.t livers, electrospray mass spectrometer, fluorinated 3.1 Use gloves when handling the rabbit livers, they may contain pathogens, 4.1 4S.u1p.p1li-eSsyringe, capable ofmeasuring 100 pL 44.11..23 GElpopveensdort ypoe disposable pipess 44..11..54 PPllaassttiicc gcernitnrdiifnuggetutbuebses, 15 mL 44..11..76 LNiatbreolgsen 44..11.89 TFiilmteerrs, Titan nylon syringe filters, 02jm. 44..11..1110 DAinaslpyotsiacballeppilpaesst:icg3lacses syrivnolguemse.tric pipers. 4.1.12 Crimp cap autovials. 4.2 4R.e2a.g1enAptqpsumeaoquuseoAumsmsoonluituiomnoAfceatmamteon(Ailudrmicahc)e,taatpeprboyx.a2dd5i0ngpp2m5:0 Pmrgepaamremoan2i5u00m `awcaetteart.eD1i0luat1e0t0hims Lsolvuotliuomnet1r:i1c0ffloarskaa2n5d0dpilputmestoolvutoiloun.me with Milli-Q 4.2.2 (aSnoNddai2,1.u0CmOcJgaoNrfHbsoOonOad,ti)eu/m0S.ob2di5icuaMmr:bBoiWnceaatiregbh(onN2aa6tH.eC5BOgu,fo)ffeisrnot(od1a.iTu.1mBLackaverorbl)ou,nmaetteriMcacflaosk)and 44..22..43 DbirliuntgetaocvetoolnuitmreilweistohluMtiiloln,i-dQilwuatteearc.etonitrile Ethyl Acetate 1:1 with Milli-Q water. 44..22..65 MMieltlhia-nQowlater 44..22..78 F1CH'9,51(H3:M22HSpH-ecipaelrtfyluCohreomoictcaanlesDuilvfiosniiocn)acid (Aldrich) 600849 , 50EQ5.U1IPUMltrEa-NTuTrrax T25 Grinder for grinding liver samples. 55..32 CVoerntmeixfumgiexer 55..54 ASnhaalkyetrical Evaporator ~~ 6I .0 N 6.1 TT herearE enoknR owninF terfE earttehnR sciemes.ENC ~~ ES 2.0 SAMPLEHANDLING 7.1 pTehreforrabmbeidt.livers are received frozen, and mustbe kept frozen until the extraction is 8S .0 CT A8.L1IA BPRreApN TarIatOiND onAoNf A IDnternR al StaD ndardI s ZAT ~~ION 8.1.1 pPerretplauroeraooncitnatneemsaullpshtoannidcaarcdiodftaoppbreoaxdidmeadtetolyea1c2hplipvmer s1Ha,mp1lHe,,2H,2H8.1.2 W10e0imghLavtolleuamstet0r.i1cgfloafsk1.HR,e1cHo,r2dHt,h2Hea-cpterufalluwoeriogohctt.anesulphonic acid into a 88..11..43TBorian2g5i0t umpLtovvoolluumemteriwciftlhasmke,tahadndol3,mtLhissoifs tthheessttoocckkssttaannddaarrdd.andbringto svtoalnudmaredw.ith Milli-Q water. Calculate the actual concentrationofthe actual mg perfluoroctane__sup0h.o1nLicacid X 235m0LmL= actual concentration, ppm 8.2 8P.r2e.p1arPereFpCar-e95FC-A9n5iosntanSdtaradnsdfaorrdtshe standard curve. 8.2.2 RWeeciogrhdtshpepraocxtuiamlawteliygh1t0.0 mg of FC-95intoa 100mi volumeic flask. 88..22..43DiBlruitnegtuhpetsoovloutliuomnewiwtihthdidliultueteacaecteotnoniittrriillee1.:10for asolution of aapsporlouxtiiomnaotfealpyp1r0o0xp.pm1.0 pDiplmu.te this solution 1:10 with dilute acetonitrile for 8.2.5 5U.s0eptphme,101.p0ppmpsmolauntdion50t0o mpapkbe., working standards with values close to 8.3 8P.r3e.p1arWeeiBgehef40LigvoefrBoHvoimnoeglievneraitnetofao25U0smeLfoNralSgteannedbaotrtdlsecontaining 8.3.2 A20d0dm1LmsLMoiflltih-eQswoaltuetri.onGrtoinad1t5omaLhocemnotgreinfuoguestusboel.utPirone.pare a totalofeight s1usmpenadlisqoulouttsioonf tbhye sshoalkutiinogniitnbe1t5mweLenceanlitqruioftus.ge tubes. Be sure to re. 6006350, 8.3.3 SStpainkdeasredsveinn osftetphe9.112moLfatlhiequportoscewdiutrhe.thOenfeoll1omwiLngalaimqouuonttsserovfeswoarsktihneg blank. (AWpoprrkoixnigmatSetaCnodnacrd.) "cAopnpcreonxtirmaattieon ofifnal 1 _ FCB-9a5 ikn liver | 8.4 Calculate the actual valofuthee standards: uc 1L71oofmsgn talnic vdearr'e d/ x1 n ml t homrogeanat te i (inoppnm) = final concoefnFtrCa-t9io5n in(plipvme)r 1*A'vmeLrhaogmeowgeeingahtteosoffbo4v0inmeglilvievreirninso2l0ut0iomnLaosfdMeitlelrim-inQewdatbeyr.weTihgehianmgount of FC-95 is reported as equivalentsof FC-95 potassium salt. 8.5 8Ca.l5i.b1ra`Etmxetitrohanocdt.tUhesesptihkeesdebseteafndlairvedrshtoomeosgtaebnlaitsheyfooulrlocwuirnvge9o.n13th1e0 m9a.2s3softhis 8.5.2 oAsfplettcehtmeraoptmeiervtfeellruy.c,raooscttaanndsaurldfocnuartveeamnaioynbaengdetnheeraitnetdemuaslinsgtarnadtaiorsdoafnrieosnpvoenrsseuss concentration of the perfluorooctanesulfonate anion. 8.6 8S.t6o.r1ag`eNceoCvwoernsedtdiapntldiaasortndissc caferonertprriSeftpugaaernedtduabwreidstshunetialchthaenaalnyasliyss.isSotn tahenmdaasrasessrtpoedrcetsdromineter is performed. 8.7 S8t.o7r.a1geBeCconlidvietrihoonmsogfeonratSetsamnadyarbdesfrozen after preparation. 9.0 PR9.O1CEODbtUaiRn fEroSzen liver samples. In spent tissue, note that the liver has not been 9.2 Upascekaagdeisdsewcittihngotshcearlpteilssauensd.cut off approximately 1 gofliver. 99.134 9.5 RWeeciogrhdtthhee Puta label slaimveprlewediigrhecttliyn itnhteosatutdayrendoptleasbtoiockg.rinding on the vial with the study number, weight, tube. rabbit ID, date and analyst initials. GOGSSL 99..76 GArdidnd2.t5hemsLasmpwlaet.erP.ut the grinder probe in the sample and grind for about 2 minutes, until the sample is a homogeneous solution with no large chunks. 99..98 TRainkseetthheegprriondbeeroafpfaritntaontdheclseaamnpiltewiwtihthme2t.h5amnLolsafwtaetrereaucshinsgamappliep.etF.ollow 9.10 AMDT-EP-22. Cap the sample and vortex for 15 seconds. 9:11 iPnifpoerm1atmioin tasothe1g5rimnLdicngentvuibfe.ug(eSebeeA,MLDaTbe-lMt-h4e cWeonrtkrisfhuegeettufboredwoicthumtehnetiidnengtithcea.l 9.12 remaining steps.) Spike thebeefliver homogenates with the appropriate amount of FC-95 standard 99..1133 aSspidkeesctrheibseadmpinle8s.3a.ndbeefliverhomogenates with 100uLof intemal standard. Add 1 mL of the sodium carbonate/sodium bicarbonate buffer and 1 mL ammonium 9.15 Uacseitnatge.an analytical pipet,add 5 mL ethyl acetate. 991:1176 PCuatpthheemisanmpltehe sahnadkevrofrocrx2200m1i0n.30 seconds 9.18 Coenntmhiefcuegnetfroifru2g0e 1t0o.2255. minutes, until the layers are well separated.Setthe power 9.19 RmeLmgorvaedu4amteLdsgolfastshpeipteotp.oTrrgaannsifcerlthae ltyabaeelfrtersthhe1f5rmesLhctuebne.triftuugbe with a 5 9.20 Baplporwoxtihmeastaelmypl3e0dtoow4n0ominntuhteesa.nalytical evaporator to near dryness with nitrogen, 99..2221 _ VBorritngexth1e5rseemcaoinndisn.g sample up in 1 mL dilute acetonitrile with an analytical pipet. 9.23" tThreanssafmeprlteheinstaomapflreetsoh cae3ntmriLfusgyertiungbeeo. rAtaiaacuhtoavi0a.l2. Ljaubmenlytlhoentmuebsehorfivlitaerl,awintdhfitlhteer 9.24 Cstaupdyanndumhboeldrfaonrdanaanliymsailsnbuymcbleerc.irospray mass spectroscopy. 9.25 Complete AMDT-M-4 worksheet and attach to pageofstudy notebook. 100V10A.L1 IQDuaAliTtIy COoNntrol- not applicable 1100..32 OPrtehceirsiVoanliadnadtiAocncPuarraacmye-tenorts-apnpolticaapbplleicable 11.0 D11A.1TANoAnNeALYSIS 12.0 A12T.1TAWCorHksMheEetNATMSDT-M4 13.0 R1E3.F1 EARMEDTN-CEPE-S22 Routine Maintenance of Ultra-Turrax T-25 1Re4vi0siRonEVISIONS Number ReafosrChoannge Revision Dae GOG352 4 EEREE E WorksheetAMDT-M-4 C95 C95 BEST COPY AVAIL ABLE C95 "Date and FF" ma t FF F----F--r ----f------+------ F rF + tr Fr F tr r r t mse eFix m ---- De flumk --| patie -- FmoeestLukof0E.h23NaCOM28MWHORHF -- bem e m= m z EE -- | . poemtdiien2028m0 |-- Plo down-- topeatdrv-- pessPOR-- aL M-- o ----------| bm 6006553 3M Environmental Laboratory Method Analysis of Rabbit Liver Extract for Fluorochemicals using Electrospray Mass Spectroscopy SOP Identification Number: AMDT-M-5 Adoption Date: (-( 45 Revision Number: 0 Revision Date: None Author: DaveChristensor/Cynthia Weber Approved By: up Leader 4 : Quality Assurance: /4, Dae ch tee Date Software: MS Word, 60 Affected Documents: M-4, ExtractionofFluorochemicals fromRabbitLivers 600854 lLescoPE 1.1 Scope: This method is for the analysisofextracts of rabbit liver or other tissues or ifslupiedrsfoforrmefduobryoscihnegmleicilons muosniintotrhiengeloefcFtrCo-s9pr5aaynmioans,sMs/peZc=tr4o9m9e,tetrh,eTihneteamnadllysis 1.2 AstpapnldiacradbMle/ZCo=m4p2o7,unadnds:otFhleruoarpopcrhopermiiactaelsmaosrseosther fluorinated compounds. 13 fMlautirdsi.ces: Rabbit Livers (samples), Beef Liver (standards), other tissues and 20KEYWORDS 2.1 Fluorochemica l s , fluo ri nat ed co mpo un ds , elect rospr ay mass spectroscopy, mass spectrometer, rabbit livers. 3.0 PR3.E1CAUUsTe cIauOtiNonSwith the voltage cable for the probe. When the voltage cable is plugged 3.2 DinotontohterpurnotbheeDpOumNpOaTboTvOe UitC'sHcaTpaHcEitPyoRfO4B0E0,0tphseir.eIifsprriesskosufreelegcotersicoavlesrh4oc0k0.0 psi isinodpathnedplluegaansdprreespsluarcee.hTehpelpugagkedtufbuibinnggm.aSyecbeAMpDluTg-gEedP.-T1r5oubleshoot bck to 3.3 Do not run the pump to dryness. 4.0 SU4.P1PLISuEppSlAieNs DMATERIALS 44..11..21 FNliutorfoigxencoglausmrnegourlaeqtueidvtaole1n4t.0 psi. 41113 100uLor250 ul fla tip syringe for sample injection. 4.2 4R.e2a.g1enDtislute acetonitrile mobile phase, dilute acetonitri.le 1:1 with Milli-Q water. 4.2.2 Milli-Q water, all water used in this method should be Milli-Q water. 50EQ5.U1IPVMGETrNioT2000Electrospray Mass Spectrometer or equivalent. 55..32 SIpSeCctOraSpyhryisnigcesPAuSm3p00 Autosampler 54 5.5 A1u80t0ovAisalsseomrbcalpyped centrifuge tubes. 6I 0 N 6.1 T There aE re no knR own intF erferencE es at thR is time.ENC = ES 2.0 SA7.M1 PLKEeeHp AtheNexDtrLacItedNsGamples in capped 15 mL centrifuge tubes or in capped autovials until ready for analysis. 600355 , 8.1 8Pr.e1p.a1raSteivoenn boefefClailvierbrsatatnidoanrdSstaannddoanredsblank beef liver are prepared during the extraction procedure. (Sec AMDT-M-4, section 8.0) 8.2 8C.a2l.ib1raRtuinonthe seven beef liver standards twice, starting with the lowest standard 8.2.2 to obtain the standard curve. Typically one standardisrun after each 5 to 7 samples. Choose a standard in the same range of concentration as the samples. 8.3 8S.t3o.r1ageFreCsohndstiatnidoanrdss faroer pSretpaanrdeadrwdisth each analysis. Standards are stored in pceorvfeorremdepdl.asStaicmpclenetsriafnudgesttaunbdeasrudnstialrethNe OanTalryesfirsigoenrattheed.mass spectometer is 8.4 8S.t4o.r1ageBeeCfonldivietrihoonmsogfeonrateBseemfayLibveefrroHzeonmaofgteernparetpeasration. 9P 0 ROCEDURE... 9.1 9In.i1t.ia1l SSeetts-oufptware to "Operate on", Ion Mode ES. 99..11..32 FRielcotrhde sboalcvkeinntgcyprleisnsduerrewiinththmeobinislterpuhmaesnet.log, 911. `Sceotmtihnegpouutmpofttoh"eRtuinp"o.fStehte tphreobfel.oTwhteo p1r0e0s0suurLe/msihno.ulOdbbseeravte1s7d0r0optloet1s800 si. 9.1.5 Check th fused silica at the end of the probe. Use an eye piece to check for cchuitposf.ftThhee itipp osfhothueldsibliecaflwait twhitahcnoolujmanggceudtteedrgaensd.Ipfulalntyhechsiipliscaartehrfoouugnhdto 9.1.6 tChheeacpkpryoopurriantietrloenggetnh.supply. Tum on the nitrogen. There should be no nitrogen leaking around the tip of the probe. A fine mist shouldbecoming. 9.1.7 oCaurteoffultlhye gtiupi.de the probe into the opening. Insert it until it won'tgoany 9.1.8 fGarothteor.thCeon"nEdeicttort"hepvaoglet,aagnedcasebtleIotnoiztahteipornobMeo.de to ES', and the 9.1.9 aIfpiptriosprnioattienmsaisnsgelesitoon4m3o7daendgo49t9o."Option" and set SIR. 9.1.10SthuertloAgcqbuoioski.tion. Assign a file name, MO-DAY-YR + letter. Record itin 9.1. 11bReugnintnhienbgeeoffltihveerruns.a.mRpluensafiQrsCt, cruhnencikngbyearcuhnnsitnagndoanred sttwaincdeaartdtahfeter every 5107 samples. 9.2 9M.a2n.u1alDrIanwje1c5t0iounL of sample into a syringe. Inject the sample into the rheodyne 9.2.2 Tinujemcttihoenvpaolrv.eI1n0je"cOtns"l.owly. Record the sample ID in the log book. 99..22..43 WRaeictortdwtohmeisncuatnesn,uamnbderinfjoecrtetahcehnseaxmtpslaempilne.the logbook. 000356 5 9.3 9U.s3i.n1g StehteupAustaomspalemptlraeyrA, B, orC. 9.3.2 vRieaclosrmdatyhebesaimnpelaecsharnudn.their positions in the instrument log book. Up to 17 9.3.3 9S.e3t.u3p.t1he samplPeurs:h the sample bution 9911331133132 SSeettsianmjpelceuslaomopplsei=ze2= 100 ul 991133:.33'45 NSaemteCtyhceleftilie:meLi=v0ers 99..33..33.76 AIdddenitfhye sthaemptlaeysutsoeQdueue by pressing "Enter" 9.3.3.8 Press "Run" to start 10.0 V10A.L1 IQDuaAlTitIy OCoNntrol 10.1. T`Rpueank haesiagnhtdaorccduvresrsytoSpt1h0e7rusna.mplOenlsy tIhfe siagmipflecsanbetfcorheatnhegleasSt 0ac%c)epitnable Standard will be used. The remaining samples will be reanalyzed. 10.2 P1r0e.c2i.s1iSoeneaMnedthoAdccVualriadcaytion Report number AMDT-M-5.0.V1 1100..43 ROetfheerrtoVMaeltihdaotdiVoanliPdaatriaomneRteeprosrt Number AMDT-M-5.0.V1 11.0 DATAANALYSIS 1111..12 PClaoltctulhaetisotnasndard curve, using the mean of the two values obtained for cach standard. 11.2. 1Read peak heights or areas for the samples from the printout. Use linear 11.2.2CreaglrceuslsaitoentthoedmetgeromfiFnCe-t9h5e asnaimopnl,eorcoontcheenrtrfaltuioornosc.hemical in the total rabbit liver: `mg FC-95 anion in the total rabbit liver = mggmEsCo-f9l5iavneriounsfedrofomrsatndal.ycsuisr.ve ~~ x Total mass of liver, gms 1111..43 PMrainkteaa crhersoulmtasttoagbrleamanfdorecntaecrhist aimnptlhee,swtiutdhy tbhoeokp.eaks labeled with the sample or sSttaunddyafrodldIeDr.. WSrtiaptleetahlel scthurdoymnautmogbrearmosnttohgeetphreirntaonutd,niunimtbiael,rdpaatgee,s.and put it in the 0600857, 1A 2.0 TTACHMENTS None 13.0 REFERENCES 13.1 AMDT-EP-17 140 REVISIONS RNeuvmisbieorn Reasonforchange DReviiseion 000858 3M Environmental Laboratory Method Analysis of Fluoride UsiTnogn tSheeleSctkiavlearElSeecgtmreondteed Flow Analyzer With Method Identification Number: AMDT-M-8 Adoption Date: /p-s-45 Revision Number: 0 Revision Date: None Author: DebWri/gCyh nthtia Weber ApprovedBy: up Leader e4l BAS Quality Assurance 2/5/75 B Dae I-27 Dat Software: IBM MS Word, 6.0 Affected Documents: AVDTRS25 Operation and Maintenance of the Skalar Segmented Flow Analyzer 600559 | oscope 1.1 This method is for the analysis for fluoride, thermally extracted from samples using the Dohrmann DX2000 (AMDT-M-1), and collected in TISAB for analysis with an SIoengmSeelnetcetdivFelEolwecAtnraoldeyz(eIrSEw).itThhIeSEa.nalysis is performed using the Skalar 1.2 Samples can be tissues, serum, biological material, or other materials extracted on the Dohrmann. 20KEYWORDS 2.1 Skalar, segmented flow, fluoride. 3.0 PRECAUTIONS 3.1 Follow standard laboratory safety practices. 4.0 SUPPLIES AND MATERIALS 4.1 4S.u1p.p1liSeasmple cups, 4mL plastic cups with caps 4.1.2 Autopipets, oxford or equivalent with plastic tips. 44..11..43 PCaorltyrpirdogpeycloemnpeovnoelnutmse,trriecfefrlatsoktsh,e1S0k0almaLr Methods for components and part 4.1.5 nSuammbpelresp.refilters, Evergreen 4.2 Reagents 4.2.1 4.2.2 4.2.3 4.2.4 Brij 35, 30% S.F.A.S. Detergent TISAB 11 buffer solution: Purchase TISAB II from Orion. To 1liter of TISAB II add 2.5 mL or 100 ppm fluoride solution and 1 mL Brij. Sampler rinsing solution: Dilute TISAB II 1:1 with Milli-Q water. Nitric acid solution for decontamination, 1 N(lab grade): Slowly add 64 `mls concentrated nitric acid (HNO,) to 250 mLs of Milli-Q water. Bring the volume up to 1 L with Milli-Q water. 4.3 Standards 4.3.1 Stock solution, 100 ppm F: purchased from Orion. 4.3.2 Intermediate standard, 10 ppm: Dilute 10 mLs of stock solution to 100 mLs 4.3.3 WwiotrhkiMinlglis-taQnwdaatredr:.MUaskeepuolpytphreopfyollleonweivnoglwuomertkriincgfsltaasknsd.ards by adding the. voxoflourmdesoorfpiunmteprmmaetdeiaptiepeotrs,sttooc5k0smtaLnsdoarfdTiIndSiAcaBteadnodnditlhuetitnabglte,o u1s0i0ngmLs. with Milli-Q water. --o --Oiespm pm --[ T -- o os s| ooe memn -- deem ---- 03 is ||| 0600860, [--Tappm 7 -- T2 T----------] 50 E Q5.U1 IPSMkaElarNSTegmented F low A uto Analyzer SansTM S yst em e q ui p p == ed with ISE 000 6I .0 N 6.1 T High coE ncentratR ions ofF alkaliniE ty, chlorR ide, phoE sphate,N sulfateC o~ r ir~onE c~an cauS se interferences. 1.0 SA7.M1 PLSEamHplAesNshDouLldIbNe sGtored in polyethylene bottles. Samples should =~ be analyzed within 30 days. 8S .0 CT A8.L1IA BRPrAepTN arIaOtiND onAoNf A CDalibrR ationD StandI ardsZATION 8.1.1 Prepare calibration standards as in section 4.3. 8.2 C8a.l2i.b1raTthieonstandardsareanalyzed at the beginningofthe run. 8.3 S8t.o3r.a1geStaCnodnadridstiaornesstofroerd Sintcaanpdpaerddpsolypropylene volumetric flasks. New standards are prepared ataminimum of every six months, or as necessary. 2= 0PR9.O1CESDtaUrtRUEp Procedure 99..11..12 CPultatmhpe dflouwonridteheelpeucmtproddeecskisn,tahiereblaercstraondde schaammpbleerr.-pump tubing. 9.1.3 Tbauthm, onthe power ofthe sampler, pumps, off potentiometer and heating 99..11..45 PTuutmthoenretahgeenitn-tlerifnaecse,inctohmepauptperro,prdiiastpelabyotadnesd.printer. Make sure you turn on the interface before the computer. 9.1.6 Let the system stabilize for approximately 30 minutes. 9.2 S9t.a2r.t1inCgreaatReuansample table by selecting FILES, TABLE, and CREATE, type in 9.2.2 tPrhienntatmheesoafmtphleeftialeb,lea,nidnsperretsesdEiNnTtEheR.system table by pushing ESC, 9.2.3 PDiRaIlNtTh, GsaRmpOlUerPse1.ttTihnigss twoiltlhepraipnptrotphreieantteirneurmubne.r of samples, number of 9.2.4 sFielclotnhdessfaomrpslaemtprlaey wwaisthh,thaendstnanudmabredsr,osfamspelceosn,dswafsorhetsheasnadmpdrlieft.s. IW and 9.2.5 SFeWtt/hReUbaNsOelUinTe.cups on the sampler do not need to be filled. 00861 9.2.5. m1aSeylebcetiGnRtAhePHDIaCSH,anRdEliAnLg TPaInMeEl..SIwFiytocuh ctaontnhoetAgneatlyrseials-Ptiamnee,lyboyu 9.2.5.s2eUlsecetitnhge sCmOalNlTsRcrOeLwdrPiAveNrEfLortahned pofufssheitnpgoFt7e.ntiometetor set the. tbhseebofitnteo.mAdofjutshtetshcrebeans.eline units approximately 3/4 inch from 9.2.5. 3thCeheicnkterftahceehsicghreeswt#s3t.andard and adjust the gain,if necessary, with 9.2.6 Gwioltl onoCtObeNrTuRn.O(LSePleAcNtEoLr,deasneldectot aannaallyyssiisspbayneplr.eDsessinegleEcNtTtEheR.an)alPyrseissstThaatb 9.2.7 PtroersestutrhnetosptahceebAanraltyosbirsinPganuepl.the local menu. 99..22..98 TSeylpeectySoTuArRITDt(ionitsitaalrs)t,htehaensalaymspisl.e table which you created under 9.2.1 (or aprnedssseEleNctTSETRAfRorTcAhoNiAceLsY),SIchSo.ose running with or without the system table 9.2. 10sAefttteortshtearrtiignhgttnhuemsboefrtwoafres,asmtpalrtesthaensdamthpaltert.heMsaakmeplseu/rweasthh/ataitrhetismaemspalree:r is 9.2.11SOeKl.ect GRAPHICS, REAL TIME to view the progress of the analysis. 9.3 L9.o3a.d1ing GoatnodCOPrNiTnRtiOnLg the Data-File PANEL, press the spacebar o bring up the local menu haingdhlsieglhectttLheOAfiDle.toSebleecptriAnUteTdOaCndApLrCesUsLEANTTEIRO)N.and enter the filename (or 9:3.2 TovRiVeEw.the calibration curve, g0 to GRAPHICS, CALIBRATION 99..33..43 TToo pprriinntt tthhee lhoigwhlleevveellsccurreevne,, ppuusshh EPSRCINtTo gSeCtRouEtEoNf.graphics. Select CSEUTRTVIENGaSn.dCphraesnsgeEStChe,maandx Eyntvearl.uePtroeassppPrRoIxiNmTatSeClRyE9E00N..Go to CAL 9.3.5 RgreotssumENtoTSEERTTanIdNGPSRIaNnTd cShCaRngEeEtNhetomaprxovtaslauempblaecpketaok4,095, go to EDIT, 9.3.6 TOoUTprPiUntT.theSerleesuclttsPRgoINtoTCERONfoTrRthOeLEpPsAoNnEoLr,PSRPNACfoErBtAheRL,asOerU.TPUT, 9.4 S9.h4u.t1doPuwtnall the reagent-lines in Milli-Q water. 99-.44:2.3 ALfetterhtehesyssytsotmeimrhnasserfionrsaepdprcoovmipilaetteellyy,3t0ummionffetshe sampler, pump and otfhfesleitnepso.tentiometer. Turn off the heating bath on weekends. Leave liquid in 99..44..54 RTealkeeastehetehleepcutrmopd-edoeuctksa,ndaisrobaakrsinan1d00sapmppmleFropvuemrpn-itguhbti.ng. 99..44..76 SOenleFcrtidFaIyL,EtSu,mproefsfstAheLcTomFpuatnedrs,eldeicstplQaUyIaTndtoinetxeirtfatchee fporrogtrheamw.eekend. 10.0V1A0.L1IDQuAalTitIy OCoNntrol 10.1. 1sRiugnnifaicsatnatncdharadng(emiidn ptoeahkighheicgohntceonctcruartsi,oon)nleyvethreys1a0mpslamepslbeesf.oIrefathe last acceptable standard will be used. The remaining samples will be reanalyzed. 600862 4 10.2 P1r0e.c2i.s1iSoene and Accuracy Method Validation Report number AMDT-M-8.0.V1 1100..34 ORetfheerrtoVMaeltihdoatdiVoanliPdaatriaomneRteeprosrt Number AMDT-M-8.0.V1 11.1 C11a.l1c.ul1aTthieonsstandard curve is plotted by the Skalar software. 11.1.2Abeltltecr.alculations are done by the Skalar software. * should be 0.995 or 111.23 PWrreiptaertehespsrteuaddysnhueemtbsetroosnumtmheerpirzinetoduatas., Iinnictliauld,edastaempthleepvroinltuomuet,, awnedigbhitnsdutsoegdetehtecr swiutmhmaallrypaschkeaegteadndoctuampeenitnstoatnhde psltaucdey innottheebosotku.dyBfaoclkderu.p Malalkdeataacaonpdysopfretahdesheets 11.4 EolnetcotrsotnuidcyDdaitsak and backup disks. 11.4. 1sGtuLdPy,sawndideasl:soEldeacttaroinsiccodpaiteadiosnctoopaiefdloopnptyodtihsekStthuadtyisfsltooprpeyddiinskthfeorlaeba.ch 11.4. 20ther studies: All data is copied onto afloppy disk that is stored in the lab. None 13.0REFERENCES 13.1 DAXM2D0T0-0M-O1rg,anTihcerHmaallidEextArnaacltyizoenro-fLFilveuroride by Meansof a Modified Dohrmann 1133..32 ASkMaDlTa-rEMPet-h2o6d,s,Op#e3r3a5t,ioSnkaalnadr MMaeitnhtoednsanMcaenuofalthe Skalar Segmented Flow Analyzer 140REVISIONS Revision Number Reasonforchange Revision Dae 0008635 3M Environmental Laboratory N Method `Thermal Extractionof Fluoride by Means ofa Modified Dohrmann DX2000 Organic Halide Analyzer- Serum Method Identification Number: AMDT-M-14 Revision Number: 0 Adoption Date: 0-375 Revision Date: None Author: Rich Youngblom Approved by: Gower) up Leader /o/5/ 25 Date wl 7 Zo AL `Quality Assurance F-2Pep-- Date Software: MS Word 5.1a Affected Documents: AMDT-ExMp-a2ndaFbllueorIiodneAMneaalsyuzrerement by Meansofan Orion EA940 AMDT-EP-3 Routine Maintenance ofaModified Dohrmann DX2000 Organic Halide Analyzer j 6006864 1.0 SCOPE, APPLICABLE COMPOUNDS, AND MATRICES f1l.1uoSricdoepef:roTmhivsarmieoutshomadtrisicfeosr. thTehoepefrlautoiroidneiosftaypDiocharllmyacnonllDeXct2e0d 0in0TwIhSeAnBitsiosluutsieodn tfoorexatnraalcytsis with an fon selective electrode. 11..32 MAaptprliicceasb:leBiCoolomgpiocaulndflsui:dsF,lpuaorrtoiccuhleamrilcyaslesruomr.other fluorinated compounds. 2.0 KEYWORDS 2.1 Fluoride, fluorine, extraction, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, DX2000, fluorochemicals. 3.0 PRECAUTIONS peeeestor 33.21 GGllaasssswwaarree iasnfdraegxihlae,usbtrogkaesensgclaasnsbmeaeyxtcraeumseelyinhjourti.es. 33..43 SProelsvseunrtibzeadsegdasseasm,pplreospemracyomflparseh,ssmeadygnaesehdantodlailnlgowprtachteimcteos rderqyuidreodw.n before starting run. 3.5 Potential biohazards due to the biological matrices. Use appropriate personal protective: equipment. 4.0 SUPPLIES AND MATERIALS 44.21 CCoommpprreesssseedd HOexlyiguemn,,HHiygdhroPcuarirtbyoGnrfardeee,,rreegguullaatteeddttoo3405 PPSSL.L 44.34 QQuuaarrttzz ggllaassss csoammbpulsetbiooanttwuibte,h RTeelfilaonnceTMGtluabsisng,G-D9o4h0r5m-a0n1n2 8or90e-qu0i9v7aloernte.quivalent. 45 Orion 940999 Total Ionic Strength Adjustment Buffer (TISAB IT) or equivalent. 4.6 Sample collection vials, HDPE. 4.7 Milli-QTM water 44..98 APcotliyvsattyerdenCehpairpceotatle,s.E. Merck 2005orequivalent. 4.10 Hamilton Syringe or equivalent. 4.11 Miscellaneous laboratory glassware 50EQUIPMENT 5.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer, modified for fluoride extraction. 55..32 DIXB2M0c0o0mpsaotfitbwalree3,8v6erosri4on861.c0o0m,pmuotdeir.fied for fluoride extraction. 5.4 Excel Spreadsheet, version 5.0 or greater 6I .0 NTERFERENCES 6.1 Sample size is limited to approximately 100 ul. This may vary from matrix to matrix. 000865 2 7.0_SAMPLE HANDLING 7.1 Samples are to be handled with plastic pipettes. A new pipette is to be used for each sample. 8.0 CALIBRATION AND STANDARDIZATION 8.1 Preparation of Calibration Standards 8.1.1 The standards required for each project will needtobe appropriate for that individual project. R8.e1f.e2rTtyoppircoatlolcyol50f-or50t0hapt pprmojFecCt-.95 in methanol standards are used. 8.1.3 For rabbit serum studies, usebeef serum as the matrix. 8.2 Calibration - Overview The normal calibration is the fluoride curve (AMDT-M-2). However,ifan optional spiked serum curve is required the procedure listed below is used. 82.1 Acalibration curve for the DX2000 is generated by spiking samples with known standards and combusting them using the same methods and matrix type as the samples to be tested. 8.2.2 Typically, three replicatesofeach standard and five concentrationsofstandards will be spiked. 8.2.3 Standard curve will be plotted as Mass Spiked F (uog nth)e x-axis and Standard Mass Recovered F (ug) on the y-axis. Generate a regression curve and calculate the equation for the line a8.n2d.4thMea1s2svaSlpuei.ked F (ug) = (Amount spiked in mL) x( Conc.of standard in ppm) x (0.6004)* *EC-95 is 60.04% F therefore 0.6004 is the factor used to convert FC-95 to F 8.2.5 Standard Mass Recovered F(ug) = (TISAB volume in mL) x (Orion reading in ppm) 83 Calibration - Procedure 88.33..11.1StRaurtnU2pormore Clean Cycles when starting instrument cach day. More clean cycles may be usedifthe previous samples contained high concentrations of fluoride. 8.3.2 Blanks 83.2.1 Prepare sample using the same methods and type ofmatrix as the test sample. 8.3.2.2 For rabbit studies, usebeefserum as the matrix. 8.3.2.3 Put serum blank in Dohrmann boat. Combust sample as described in section 9.0 and a8n3a.l2y.z4e Fsoarmprlaebbaictcsotruddiiensg, ttohmeemtehtoedr AreMaDdiTn-gMfo-r2a fbolratnhkesiaomnpsleeleschtoiuveldelbeect0r.o0d3epapnamlyosirsl.ower jbuedfogreemepnrotcoefedtihnegopweirtahttohrethcaelriberaadtiinogn.is Bstuamblesawmiptlhersesupnteiclt tthoishilsitmoirticiaslreraecahdeidn,gsor(purnetvilioiunst4h3e hours). b8e.3f.o2r.e5prFoocreneodni-nrgabwbiitth stthuediceasl,ibtrhaetiboln.ank readings should reach a predetermined ion concentration 8.3.2.6 It may be necessary to mix approximately 0 mg of charcoal with the sample to aid combustion. 000366 3 83.3 Standard Curve 8da3y3.(1=3I0fbmeLe)f.serum i frozen, thaw at least enough to complete the standard curve analysis for the 83.32 Pipeite 100L ofbeef serum into Dohrmann sample boat. 8qu3a.n3ti3tyoSftatrhtewistthantdhaerldoowensotrsitnantdhaermdatcroinxc.entFroartiroanb.bitUssitundgieas,Haumsiel4tounLosfysritnagned,ajredcaatndfeijxeecdt it on or in thebeef serum. 83.3.4 Atleast 3 replicates should be used for the lowest standard concentration; more replicates `may be used at the discretion of the analyst. 8833.33..65 RCuonmbaullst15thsteasndaamrpdlse. asIfdoensecrriebpeldicianteseisctsiiognni9f.i3caanntldyadniaflfyerzeentacfcroormditnhge toothAeMrDtTwo-M-2. roelpdlirceaptleisc,atreuannadnotthhaterthseanmpelwerfeoprlitchaatteswtialnldabred.usIendditocactaelciunldaatteatthheartetghreesnseiwonrceuprlvicea.te replaces the l8e.a3s3t.07.95W,hceonnsaullltswtiatnhdasrudpserhvaivseorb.een run, calculate the 12. r2mustbe at least 0.95.Ifit is not at 833.8 Anew standard curve should be run when the combustion tube or sample matrix is changed. New standard curve may also be run at the discretionofthe analyst. 8.4 Storage Conditions for Standards 8.4.1 Storage requirementsfor standards are dependent on the individual standards used. Typically, standards are stored at room temperature in plastic screw top bottles. 8.4.2 New FC-95 standards should be prepared at least once a month. 920 PROCEDURES 0000 9.1 Typical Operating Conditions: 9.1.1 Combustion tbe temperature = 950C. 9.1.2 Oxygen and Helium flow = SO cc/minute. 9.1.3 Vaporization/Drying time = 240 seconds. 9.1.4 Bake time = 300 seconds. 9.2 Start Up Procedure: 92.1 Ifthe program is not started, start the EOX program on the PC. 9.2.2 Open the SYSTEM SETUP window. 9.2.3 Put the furnace module and the cell in the READY mode. 9.2.4 Close the SYSTEM SETUP window. 9.2.5 When the oven has reached the READY temperature, run the CLEAN BOAT program found in the CELL CHECK menu. 9.2.6 See AMDT-EP-3 for detailsofthe Dohrmann software. 9.3 Sample Extraction Procedure: 9.3.1 Open the SAMPLE HATCH and pipette 100uLofsample into the BOAT. It may be necessary to mix approximately SO mg of charcoal with the sample to aid combustion. Ifthis is dstoanned,acrhdacrucrovael.should also be mixed in while establishing the baseline and when generating the 9.3.2 Close SAMPLE HATCH. 0008362 4 l9.a3b.e3ledAdsadmpaplperocporllieactteiovnolviualm.e ToyfpTiIcaSlAlBy 0s.o6lumtiLontoor151:m1 LTIaSrAeBu:seMdi.llFio-rQrTMabwbaittesrtumdiixest,urueseto1a.0 or 92..3.04mLPloafce1:t1heTIviSaAlBs:oMtihlatlith-eQTMtipwaotftehremiCxtOuMreB.USTION TUBE is in the TISAB at least 0.25 9in.c3h.e5s.RuGnastehse rEeOleXas-eWdAduTrEinRg ppyrrooglryasmisfmouusntd binubtbhleeRtUhrNoumgehntuh.e TISAB. 9.3.6 93.7 IWfhuendniltuhteeEd OTXISpArBogwraasmuissefdintioshceodl,lercetmtohveesatmhpelceo,llaedcdtiaonn eviqaulalfrvoomluthmeecoofmMbiulstliio-nQTMtuwbae.ter 10 the TISAB to make 9.3.8 Rinse the end of 1:1 the TISAB:Milli-Q7TM. combustion tube with Milli-QTMwaterand wipe with a KIMWIPE to remove any TISAB remaining on the tube. 9.3.9 Open the sample hatch and remove any remaining ash from the boat. Ash can be removed b`woittthaomcwoitttohnatsippapteudlaapoprlioctahteorrsainmdi/loarrvdeavciucue.meAd odurto.pIotfmMaiyllbie-nQeTMcewsastaerry tmoasycrbaep apadrdteicdletsootfhfethe boat to aid in the Clean Cycle. 93.10 93.11 Close the hatch. Run the CLEAN BOAT program. 93.12 Sample is ready for analysis by ion selective electrode (AMDT-M-2). 9.4 Sample Calculations 9.4.1 9.42 Use the standard curve to calculate the sample value, Sample Mass Recovered F (ug) = (TISAB vol in mL) x(Orionreading inppm -intercept) (Slope) 0.0 VALIDATION _---- 10.1 Quality Control 10.1.1 Daily Start Up Check Samples: Once the standard curve i established, cach day of caonnaclyesnitsraitsisotnarstpeidkebdy sanaamlpylzeisnugsQedCtosagmepnleersa.teTthhee sQtCandsaarmdplceursvaer.etEoabceh ctohnecsenatmreaatsiotnhemulsotwebset done in triplicate unless the first two replicates are within 20%ofthe standard curve, then a third replicate is not necessary. F1l0u.o2riPdreecNiostieobnoaoknsd 2A,c3c,uarnadc5y.: PSreeceimsieotnhaondddaecvceulroapcmyevnatriaensalwyhsiesnaanndalsyazmipnlgesaanmapllyessiosfidnifferent `matrices and different reference compounds. 103 Other Validation Parameters: NA 11.0 DATA ANALYSIS 111 Calculations 1111.112.21 TFoorctahlecusltaatnedtahred fcluurovrei,deusceonrtergarcetsisoinoninantahleyssaimspilne,Exsceecl,mevterhsoidonA5M.D0To-rMg-re2a.ter. 5 G006368 11.2 Analyzing the Data 11.2.1 2 must be at least 0.95 or greater. "Outliers" may be excluded if two of the three replicates are within 20% of each other and the outlier is greater than 200% of the average of those two or nloestsedthiannt5he0%FionfatlhReepaovretraagleoongftwhiotshethtewor.easAonnyitswuachs ocuotnlsiiedrsersehdoualndoubtelipeori.nted out in the data and 1A 20 TTACHMENTS None 13.0 REFERENCES ~~~ 13.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer Operator's Manual (Manual 915- 349, 13.2 revision B, December 1993) AMDT-M-2 Fluoride Measurement by Meansof an Orion EA940 Expandable Ion A13n.a3lyAzMeDrT-EP-3 Routine Maintenance ofa Modified Dohrmann DX2000 Organic Halide Analyzer 140REVISIONS Revision Number ReasonforChange. Revision Date 600869 6 9.3 Quality Assurance Unit Statement 600870 Attachment D GLP Study Quality Assurance Statement Study Title: Single-dose Intravenous Pharmacokinetic Study of T-6052 in Rabbits Study Number: AMDT-111694.1 NaofAmuditeor: Kari Rambo ``TThhiesfsitnuddiynghsaswebreeenreipnosrpteecdtetod tbhyetshteudQuyadliirteyctAosrsaunrdanmcaenaUngietmeansti.ndicated in the following table. Inspection Dates Eom To --haee 10/1395 101995 Final Report Date Inspection Reported to Management StudyDirector 101905 10/1995 oe br t-2y. JAU Auditor Date. 6008711 9.4 Key Personnel Involved in the Study CO6872 3E M nvirLao boran torm y en~ t~a~l Key Personnel - ThermJailmeJxothrnascotnion followed by analysis using Orion ion analyzer: Deb Wright Rich Youngblom Deann Plummer Analysisofliver extracts Jim Johnson using electrospray mass spectrometry: Dave Christenson Thermal analyzer weixtthralcotniosnelfeocltlioveweedlebcytraondael:ysls using Skalar segmented flow Jim Johnson Deb Wright "Rich Youngblom Deann Plummer Documentation and Reporting: Jim Johnson Rich Youngblom Quality Assurance Unit: Gale Van Buskirk Cynthia Weber Kari Rambo 000373 9.11 Data 000874 9.11.1 Summary and raw data; ug F" in whole liver as determined by thermal extraction followed by analysis using Orion ion analyzer. This data, although supportive, in the opinion of the Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. 606375 Summary of Combustion Data - Liver AMDT-111694.1, HWI 6329-134 As Referenced in Final Report section 6.0 DATA ANALYSIS Total ug Fluoride in Whole Liver Mean per Dose Group Control Group ~~ 2u6.g3 2.0 mg/kg dose (T6052) ~~ 17.9 20.0 mg/kg dose (T6052) ~~ 17.0 200 mg/kg dose (T6052) 35.1 1000 mg/kg dose (T6052) ~~ 77.7 600876 FC120PK 0 LLiivveerr BBllaannkk-21 LUvveerrSSppikkeet2 FUov2e5r4S8p.1ike3 FFes22554488.32 FFss22554499.21 FeF2s2855400..13 FFas2255505-023 FFs5225s6668-.12 FeF2s2556667..13 FFss22556677-.32 LLiivveerr 8Blikk--12 LUvveerrssppkkt2 LUvveerrsSppkks3 LLvveerrsSppkkss pAcPtmiFal- ApveprmaFg-e % inliver inliver ry (0W35W5) (WW) 103% 011.8161 8942%% 11.3244 0328 00230777 021 00115799 0200 00221674 0022121 0257 00327828 03 00835333 0079782 os7 00..110383 9800%% 01..91800 0942%% 21.7026 10048%% 234234 fiver Wlhiovelre bumed weight (g0r.a10m9s) (grams) 00.113450 00..110025 0108 03 00112176 90033 0o1f3011 893 803 00110274 781993 0011285 7M199 00113493 11005514 00113373 1800571 00110018 88977 00.111226 00.111309 00..110352 00.11071 TowtahlolFe-in liver (ug) 263 19 170 351 77 RPTIS4LXLS Dosage (moka) 00 20 200 20 1000 Page 1 600877 9.11.2 Summary and raw data; analysis ofliver extracts using electrospray mass spectrometry. 0006878 HWI#6320134 PrTsetouacdtoyi:lamHaunmber RfRSeasrsedpFVaaocienr:nAmosunet: VADaarnatthy:: UiAnsBtBraAmSenEtF:ie: `TfSisgilimCoosRaaniavsar(cFuCs1P2a0rmacokiesc SGNoArering aoies GFioonusasVsG2000EacospaHyS An Corkins er A heh'A 2.0D-i31n-i9n5g, lo[mtaaorreoWa+ter Seed TanArCeaa Fase ND. 9 Dnmer dw 1 Congeetnaton Toaflg ers No. soz TFososparTive of mg ross No. frre | se NO. tess 1 No. anzse No. [froemaa mw NO. tem 1 No. roan no. J[eoemwoa sss ' ' 105080 f[ooorommsa:ria rer 8 wor Fr =Posiiveresponsefo kon manfred. + sosast a.590 "{TaiocswicCooarnlacuetnlhtaertaeotnaiouen weoefrstcmahilnsccu1lla7ct4lemagtb.yaTunshnienacgpptohiennsgctfaanoduanrldisicurr6v.ah4ee1alsntaodatnmndhaaolerdtdpAlcyuMirDnvgYetAahKre4e0ha:sr1tel7boy1r4g554.fTrhegere4w5haaseueydIn <houidbe. Bymulipyig thecalculated concentation nthe sndarscurveb 45, a cectrecutlscoed. 600879 Ubese menos oct 1sBEST COPY AVAILABLE Az From Fils DSpaemrpalzeordlcd]lciu ofof"ssC Kun Pdraitente0d5-0on8-10757-530-0199:9354:5A6T Versions 09:34:14 1 Straiaht Line Fit forced throudh origin WTP. wn 13 Easiolbe PoBidopninlhbier oy res ; Usnponent #:1 i ye ne 3# |i I i A sug i re /a 5 8% | / mo sig i To 4 16 8 . 3 4 HHT 2 CEVEL amount EXTERNCAoLmpSonTeAnNtOAKD1 CA=LIBRATION AREA :1 5 0.0.45000000 1.5000 9714070379 137121 .: 45100000000 s1o0l7a0u3s6 @ 12.0000 10095 Ye SLOFE x Kx (NTERGERT mwouHnats a W suarad =16750.a50-3015Es0x s = 0.9720 x AArmeoaunt ++ 00..00000000LE+s000u 000850 n< copy 3 $32 FSiamlpel:eoHdtIssc 6329-194: FGLA1B2B8AS(EPH>= TmhoamNSaDaLtoa nSyBstooonmers MT TTaeT es Tom om sao, a [#Fs- 8 Ed oarmasises 13:50 4003g pe 1763 -rs y Ti 3 S330) boemlo yt Ts r TEN9 %i 1108 ol 2g7AJ332e A432 532 L63N5 LN usgJpieHM1231890 3sam 40349 aLgey ai0 son' 6 360 coo ude joao izes impe oa a0 v 5 < 3g cvaemrps FOLLblO ne11. E naE sstE er suis oi |108- ao 1389 402333 wh 3 { 5 g on 1TH ag Ty x ls; 0 1959 1166 1156"1760 1350 1398 1350 40 1456 15ee 135 o< agg2 SaFimlpol:eaaHaUaIssc8 6329-134; FCL'A4D2-6DAS(EPX)~ TBhaovbMiSrDaLtivaenSyEsvteeamncrs oa/pa/ioss 13:53 ey sep HR 1igr pine use . 0 Rane!i133 oo Le 110 fe we 1a naIg - brs a sen 1066 #56 1166 1150 1760 oo A 1256 "1366 024 6 \ 1356 14G Sid | CT 8 2 z2z < 2 iSieasrre rt saro-s00; PLETaEn ThT o .1 E putsStcenrs owas 13:50 wr . 20 332 432532 [sf 9cho gbe ideo 1760"i400 ices igGd d9e.1t1e.r3miSnuemdmbayrtyhearnmdalraewxtdraatcat;ionugfoFl*lionwewdhoblyealniavleyrsiass suesliencgtiSvkealelaercstreogdmee.nted flow analyzer with ion C00385 RE: 6329-134 LIVER SAMPLES AMDT 111684.1 Date of Analysis: 3-38-95 Analyst: DDL -- The samples are burned in the Dohrman at 958 C using between 0.1 and 0.2 grams of the liver. The gas is collected in 1.8 mL of 1:1 TISAB/Milli-Qwaterthen an additional 2 mL of 1:1 TISAB/Milli-Q is added to allow for sufficient volume forSkalar analysis. The samples are then analyzed on a Skalar Segmented Flow Analyzer using the lon Specific Electrode (ISE) Method. TISAB buffer is added to each sample as it proceeds through the system. The sample then goes through a heated mixing coil before the potential between the ion selective electrode and the reference electrode is measured. The signal is amplified and related to the fluoride concentration. The instrument was calibrated in the ranges of 8.815 - 8.15 ppm high and 8.15 - range was p1.l5o0ttpepd musfilnugortihdee. inTvheerssetalnodgaarrditchumrvoeptfioonr.tThhee standard curve for the low range is linear. All standards and samples were then calculated by the Skalar softwareusing these curves. All results below 8.8881 ppm appear on the raw data as #.####. A quality control standard was analyzed every 10 samples to check for accuracy and drift. Raw data is taken from the appropriate calibrated range of the Skalar printout and summarized on an Excel spreadsheet. The final results are adjusted for the collection volume and any subsequent dilutions. Oruigs CL Ntef, is C0G386 SUMMARY OF 632-134 LIVER SAMPLES AMDT 111684.1 DGcrlooudri1o EFeSlS MN3300 oollwe M Np w sno0u5s ww DelGordoiuzeis FPaessaa oNoMm Passa oo 3300 30 oownees ome oM s w om bowws was w us w 3s DoclGerdo:wNsmple BEaassssaa 0W0s30030 Pass M30 oowhmn 0w3w one ww mnmswnw noms Ww DoeloGcr:ouMponge EFussees: 002 3300 o01n6 o0s3 ok sooswo a55 a DolGor:ow10s0mg EBa2se2d 0000s Es03 0% 3300 oonum L1w% 30 owm Li ii eessse w1hm ass 0s 6 2 e ` 22~ zi 3 b Tess Paget 2RO ens DLoomtvee iuo oonnaeomoe L Si oEs mtuswm oSonme mm om S LSo oo oSsad Goommn oooomnn iwemem W BFR oS os oolm oaam woow BESTCOPY AVAILABLE B BWoooa ots maokaandoae wm Wbbooo mmmaannn Wwooowr BB20 ooowmn ootawo FFbRRo ma xoonsoss FR on BSBo aanomnee iinn oon lw cooommm ooGmee oaons onoaee lowikmm N35e9haaas Bm owwo ae moe BBBo oolowwan iWWwwe wwmw owwo wom on om me EFFRaa aooone 0Baoew owno wwamx ousw A%Baa rsae ava0inss B3Fr oonmw oWnw npmmee ow E5 oun mrhamnme oa0onr FBBri oohn Ww on nam mem gu mal & oss Boo ih wes ow Paes v5raTsa we aon ntue w oGFR oeam we aoioer ne sins em ux mew ow ew uw mew ws ggge X2 a02 1995-03-30 13:29 Output of : 95033081 Software : version 6.1 c1990,93 Operator ddw Date of the Analysis : 1995-03-30 11:18 Analysis File Name : C:\SKALAR\DATA\S5033081 Fluoride 1.5 Calibration order = Inverse Logarithm Slope BEER R 8x -c1e Result = 108 448s484 i x = corrected value of the sample cl = corrected value of the concentration 1 4 s = Slope of the electrode a2 = -0.00000 aa0l == -01..0103028458 Fluoride L Calibration order = 2 Correlation : r = 0.99665 Result = a2 * x + al * x + a0 a2 = 0.00000 aal0 == 00..0010601351 sampler Type + SA1000 NSuammbpelre Time :i 150 sec. Wash Time : 120 sec. . Air Time : 1 sec. sTpaekceiaulp :: SNionngele needle Height : 70 mm. Diluter needle Height : 80 mm ddiilluuttiioonn VFoalcutmoer :: 21.05 ml. RDeislaumtpiloen runs :i 11 User file : . TXT Reproces : No 606390 1995-03-30 13:29 output of : 95033081 Fluoride 1.5 SPiagtnhalnutmybpeer :: De3bubbled DesycsotleonrNumber :: Y0es dReisLaumtpele : No `No dil Threshold : 4095 . WdiinGdoowuetvpenutt:: O0ff 5s21 ssTTaannddaarrdd :: IIggnnoorree 5834 ssTTaannddaarrdd :: IIggnnoorree 5556 ssTTaannddaarrdd :: Ign0o.r1e50 8878 ssTTaannddaarrdd :: 00..360000 85910 ssTTaannddaarrdd :: 11..520000 DOridmeernsi: onInv: erPsEeM Logarithm sttrairgtgerValLuiemit :: 1508000 SDeUc sPteaarktshiagpnoere :: 6P0ointeSedc MeeNadsureignwoirnedow :: 71520 S%ec FRielgteenreration :: NNoo : ofourtnUplaut:: #h.E#E Fluoride L SPiagtnhalnutmybpeer :: De0bubbled DesycsotleonxNumber :: N0o dRieLsuamtpele :: NNoo ddiilG Tohuretsphoultd:: 40095 Window event : Off C0689, 1995-03-30 13:29 Output of : 95033081 5s21 ssTTaannddaarrdd :: 00..003105 ss43 ssTTaannddaarrdd :: 00..009600 s565 ssTTaannddaarrdd :: 00..112500 8587 ssTTaannddaarrdd :: IIggnnoorree $8910 ssTTaannddaarrdd :: IIggnnoorree DOirmdeenrsi: on2 : PEM sttrairgtgerValLuiemit :: 1580000 SDeUc sPteAarktsihgapneore :: 6P0ointeSedc MeeNadsureigwnionredow :: 17205%Sec RFeigletneerration `: NNoo . ofourtmUplaut:: cH4.:4=4c#3E 606392 1995-03-30 13:29 outPut of : 95033081 FFlluuoorirdiedIe1.5 Ppooy Pos Typ Ident Dil Weight Ch Result F Cor. Valu Time wt iw Initial Wash 1 1.000 33 0.00.106714 o0 128To esTo 1c Tracer 1 1.000 43 0.18.8274:2 22116677 23130 2120 2a Drift 1 1.000 43 0.19.027724 22220077 23700 3880 3 ow Wash 1 1.000 34 00..0017641 0oo181 578To 4 s1 standar1d 1 1.000 43 0.00.01795 1111 1920 7470 5 s2 standar2d 1 1.000 43 0002.08758 3300 2120 9110 6 53 Standar3d 1 1.000 340.00.s099a 113377 3200 10860 7 s4 Standar4d 1 1.000 340.00.818133 222266 4100 12640 8 85 standar5d 1 1.000 430.01.215378 333388 5240 14370 9 86 Standar6d 1 1.000 320.01.416553 339999 5860 16120 10 7 Standar7d 1 1.000 43 0.03.25819 779977 9920 17870 11 s8 Standar8d 1 1.000 340.05.167015 11336622 15680 19630 12 89 Standar3d 1 1.000 320.18.825369 22116633 23860 21370 13 510 Standard 10 1 1.000 431.10.345656 22146600 26940 23120 14 a Drift 1 1.000 320.15.331173 22225%6 24400 24870 5 ow Wash 1 1.000 43 0.00.106714 o0 184To26o68 Page 1 of : 606893 1995-03-30 13:29 output of : 95033081 FFlluuoorridiedLe1.5 pPeex Pos Typ Ident Dil Weight Ch Result F Cor. Valu Time 6u BLK 1-A 1 1.000 330.06.202707 1159 2000 28360 17 wu BLK 1-8 1 1.000 340.R0b0as1eAn 3-359 1380 30120 8 ou BLK 2-A 1 1.000 43 0t0o0o78l> 2-277 1480 32420 9 wu BLK 2-B 1 1.000 43 0000.02658 4-343 1280 33600 20 u SPK1-A 1 1.000 43 0005.03902 111177 288o3o 536 21 wu SPK1-B 1 1.000 340.00.50127 111133 2800 3714oo 2u SPK2 1 1.000 3do0.00.s08915 113333 298o3o 888 23 u SPK3 1 1.000 43 0.00.s05954 112266 2880 40590 2u FS2548-1 1 1.000 43 0001.02702 1-313 1450 42360 25 u F52548-3 1 1.000 340.A0b0s9eAn 2-233 1320 44120 2a Drift 1 1.000 23 0.15.121072 22220166 2368o4o 588 27 w Wash 1 1.000 43 0.00.106714 0o 145o4o 771 2u FS2549-1 1 1.000 13 0.00.00774 33 15270 45040 29 nu FS2549-2 1 1.000 43 0A.b0s0e9n8 A 2-222 1280 51130 30 u F52549-3 1 1.000 d3 o0.00.007535 <-22 148o5o 285 31 ou F52559-1 1 1.000 43 0.0.00703 7-7 1440 se610 Page 2 of . . | 000894 1995-03-30 13:29 OutPut of : 95033081 FFlluuoorridiedLe1.5 PPooyy PosTyp Ident Dil Weight Ch ResFuClort. Valu Time 32 u F52559-2 1 1.000 43 000.10723 3-3 1480 5606 3 ou F52559-3 1 1.000 43 0A.b0s0e6n6 A 3-311 1200 58130 34 ou F52566-1 1 1.000 34 0.00.108765 88 16"0 o59o82 35 wu FS2566-2 1 1.000 340.00.208728 2266 1780 6163 0 36 u P52567-1 1 1.000 43 0.00.502902 111144 2650 63380 37 uw. 38 a P52567-2 Drift 1 1.000 43 0006.02827 8833 2360 65130 1 1.000 43 0.18.929682 22119522 23450 66890 39 Ww Wash 1 1.000 33 0.00.106714 o0 154To68o71 40 u F52567-3 1 1.000 320.00.a02827 883 236o7o 038 aa Drift 1 1.000 43 0.19.122822 22221177 23680 7213 a2 ow Wash 1 1.000 340.00.106714 0oe 150To74o50 WE rw RunOut Wash 1 1.000 34 00..0017641 00 152o7o 688 Page 3 of . . 600395 I m-- 3 od > wp : 3 . o 5a aw 5 9 60. RREE Eac-EXIt | FiHalp | Crel-Pecdit peaks A wo7 emng - 00396 a date or oscEIDS Fivariee 1.5 1nl i 2 *f |l wis 2 RA X es | LAr E=T = T STE aT Te x oe 000397 | a dt or oscaFiavo0riane1s.3 pn 1g 3 Ag, \sz er e788 eT Tine 11173 Gone Nglas 000898 Sattar sures or secasces + Pimsrite | } Order : 2 assured ri o.seees coosss r---- o-cr\ eraes J / . order Inverse Logarithm" a: am O 600500 ! 9.11.4 Summary and raw data; ppm F~ in serum as determined by thermal extraction followed by analysis using Orion ion analyzer. This data, although supportive, in the opinion of the Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. 6006301 AHWMIDT632191-1163944.1 DAnoahlrymsiasnnDaSteesr:u0m7/A3n1a/l9y5si-s08/2/95 DAlXl2s0e0r0umOrsgaamnpilcesHawleirdee tAhnearlmyazlelry aenxtdrcaocltleedcbteydainmoad1i:f1iemdillDioQhrwmaatnenr and eTxIpSaAndBabsolleutiioonn.anTalhyezesra.mpTlehse wDeorhemmaeansnurweadsocnalainbrOarteidonusEiAng94304ppm, 0ThpepmOr,io6n2pwpams,ca1l0i0bprpatme,d b12y4dpiprmec,t2m5e0apspumr,emaenndt5w0i0thppnmo FbCla-n9k5csotrarnedcatridosn. uSlsoipneg,0i.nt0a5rcpempt,,0a.n1dppcmor,r0el.a5tpipornm,we1r.e0prpemcoarndded1i.n5pthpem aFp*psrtoapnrdiaartdes.IogTbhooek. A2).suTmhmearsyumtmabalreyistianblceluadlesdo,sshhoowwsintghethaectpuaplmOFr"ioinn ereaacdhinsgasm.pleAn(scieniptaiagle tcahleiTbirnaetiiosnocnupravegewsit3hasntdan4.dard deviation, %RSD, R? value and equation of Psaagmepsle5sawnedre6bsehionwg athnealeyxzceedl.spreadsheet that was generated when the The Dohrmann FC9S calibration curve was not used to generate the data. Quon AR an FY mins Page 10f6 006302 STUDY# 6320-134 SERUM FC120 PK HW 6329-134 Fluoride concentration in rabbit serum (ppm F-) Sample _2hour hour hour 8hour 12hour 24 hour 48 hour Dosage: Omgkg F52548 0453 0359 0334 0411 0383 170 0678 2mgkg F52540 0372 0320 0322 0384 0353 088 0642 20mgkg F525 0357 0283" 0532 0291 0200 0584 0594 200mghkg F52566 0400 0282 0539 0438 029 0564 0718 1000mgkg F52567 0402 0367 0427 0519 0327 0684 0560 Dosage: Sample Omghkg F52548 2mghkg F52549 20mgikg F52559 200mgikg F52566 1000mgkg F52567 Actual Orion Reading (opm F-) Zhou 4howr Ghour 8hour 00228 00180 00167 00208 00188 00164 00161 00192 00179 00142 00266 00145 0200 00141 00270 00218 00201 00183 00214 00260 12hour 24 hour 48 hour 00191 00848 0.0338 00177 00340 0.0321 00145 00292 0.0267 00148 00262 0.0359 00164 0.0342 00280 606303 2 NORMANSERUM CURVE 1 Saampplee mmesdnN g| GvyG gfSaSAvAeSS LT sCSpeOwdSTG CoEu3t4FCeOrS fCeTcoosvey pepAmnFAeesMGperadyregMceoswsed 14% Ou Gifs STOEV. ootsez SUsomnnnsE Dpmi 0GOo0lmeawis 0o01i1 00% 01 232000 ooGooodso 30 Gos 3344 4400 tfaammee same 111a28 175% 0ooGaewy OoGe1 O0n1e1sm 4RD: Ors STOEV. 2 0o2ucsy ape 13s ooeer Oisss RSD: 4 oLmm fommnmi meee b6i1 pOeRe 0611 2200 oooo 3300 Ocooow 4400 o@e f20w% ows ttizs lam oOoostt oonwarey Oued Ouse STOEV: 000s wot seo ome ore ws: Se SaomEmnEsi low? mO0U0wR2S G011 01 Ole a1 Z20o 30 odoow ood 20 ood M0sb os7o9w% 2il0sa0eo Oouds;e Oz: o0r1s8a4r 020% SATSODE:V oozed Wo ow 200 oz; 0mm omwememean?t fewmd lOOeFIw 060111 OWT G1 323000 oGooosw Zo oo 1ii.ma 20 ieoo mm 32202700m0 ooozmzsm 0o03aa ASTSODE:V 00sa 0% casa) 0604 Oe STOEV: 00ER072 ZZowwmd ZSoopommdl O0y% o8n1 OOMMe 0611 2200 oooomd 3300 oooodd m20 0s0 f2o%m Tew 4G1Ms)0 Bao 0Om0on0d 1208 0O4eeiesse wed Ose STOEV. fr oodsuts 7ee oe fae ice sD % Sopmd Die 01 20 cod so aw sre tans osTme SERUM CURV1E NORMAN (1772598 os yeeagevsome a 33.or .: - i - . a. 3 a doom ES 2) oosssDwrio) To + : To Te 000304 3 SERUM CURVE NORMAN (0725758) yeozacom : os : E @ w 7 ggg Gm ES) oem asorr3 Toon To Ton x STUDY # 6326-134 SERUM 8 Sample Aciial reading Sample Gly TISAB iMLFCOS fnalvol spked ONGFCO solution 1 %1 Actual | recovery ppmF- Mass spiked Mass recovered 1D BBLLAANNKK2 (pmF) (org) (mb) 0000381340 00.11 2200 (gpm) (uglug) insample (ugF) 766 (ugF) 0.168 0627 0.0827 SSPPIIKKEE6G2212 0000683454 0O011 2200 o00o4 6622 8886%% 112278 001155 00412287 SSPPIIKKEEZZ5S00-12 00213507 0014 2200 00000044 225500 5820%% 437158 006800 00341758 FSSPI2K5E4Z82.520-H3R 000224206 0O.011 2200 0004 250 80% 0448803 060 00.4048503 FFss2255459922HHRR 0000117886 0044 2200 00337527 00..00337572 FFss22sse66722HHRR 00.00220010 0044 62PPM3 00560 0.1 2200 00..440020 00..00440020 20 004 62 78% 142 015 0112 62PPM4 250PPM3 00834 0215 01 01 250PPM4 0240 O01 20 20 0004 0004 62 250 17122%% 412607 001650 00146279 20 0004 250 80% 480 060 0480 BLANK BLANK2 ~~ 00722 00386 01 0. 20 20 BLANK3 00317 01 20 0174742 00..0174742 0633 0.0633 BBLLAANNKK4S ~~~~ 0000234470 001 SPIKE621 00658 0 2200 20 ooc4 62 00455830 00..00460830 88% 132 015 0132 SPKEG22 SPKE2501 00802 0200 O01 01 SPIKE2502 0204 01 20 20 0004 0004 62 250 108% 160 6% 400 015 080 0160 0400 20 0004 250 68% 407 060 0407 SPIKE2503 SPIKE250-4 0283 0.185 01 0.1 BLANK 00788 O01 20 20 0004 0004 250 250 8614%% 530679 006600 0030679 20 1.50 0.159 BLANK Fo25484HR 00237 00180 01 O01 20 20 Fs25494HR 00164 01 20 00345795 00..00437559 0329 0.0320 FFss22S5e5e044HHRR 00.00114421 0011 2200 Fs2s674HR 0.0183 04 20 00..228823 00..00228632 0.367 0.0367 FFss22554489.66HHRR 00.00116617 0O11 2200 F52550.6HR 00266 0.1 20 00.332%2 00..00333242 052 0.0532 62PPM1 e2pPM2 00842 00722 01 01 250PPM1 0101 01 20 20 oo4 0004 62 62 7% 9% 110484 001155 00110484 20 ood 250 33% 201 060 0201 22s500.PPPPMM23 00215519 0011 BLANK 0786 01 2200 0000044 225500 5s%a% 530137 006600 00530137 20 15.7 157 FF5s2a5s6e6r.66HHRR 0000221740 0041 20 20 053 0427 0.053 0.0427 0411 0.0411 FS25488HR Fs2549.8HR 00206 00192 01 04 20 20 F52550-0HR 0.0145 04 20 00.239814 00..00239814 G00306 STUD#Y 6326-134 SERUM 9 Sam10ple FFosz2s5o6o7o8nHRR FF5s2zss448-g122HHRR FFSs22556560--1122HHRR Fs622seP7P-M112HR 2e520PPPPMM21 250PPM2 BBLLAANNKK BLANK BBLLAANNKK 6622PPPPMM12 250PPM1 225500PPPPMM32 225500PPPPMM.45 BBLLAANNKK FF5225544892244HHRR Fs2s50-24HR FFss22s5e6672244HHRR FFSS22554408.4488HHRR FFss22S56560-.4488HHRR Fs2BSL6A7N4K8HR BBLLAANNKK BBLLAANNKK 6622PPPPMM12 250PPM1 250PPM2 Actual reading (omE) 00..00221680 0000119717 0000114485 0000176444 000298145 0339 0000447815 00295 0000330014 0000850833 0.185 00240715 00222487 000120055 0000834480 00282 0000238422 0000332319 0000230579 00280 0000631%3 00383 0000431129 000073846 0304 0269 SaQmtpyle (miorg) 0011 0011 0011 0011 OOff 01 00.11 01 O0011 0011 01 0011 0011 0011 0011 0.1 0011 0011 00.11 01 0011 01 0O041 0011 01 01 TfiInaSivAoBl 'MLspFiCkeSd5 (mh) 2200 2200 2200 2200 0004 2200 0004 0004 20 0004 2200 20 20 20 20 ogo 20 20 0004 0004 2200 00000044 20 20 0004 0004 2200 2200 20 20 2200 20 20 20 20 2200 20 20 20 20 occa 2200 0004 0004 20 004 OAsoGluFiConO _Gom) 62 62 22550 62 62 250 22550 250 250 6622 250 250 reco1%very (ugg) 100% 12% 191%3% 78% 16028%% 658%% 82% 76% 112058%% 19001%% pApdmaFl- | sMpaiskse'd insample (WF) 00541396 00335833 00.2299 0.134297 015 18 568 015 060 679 080 00.94720 05% 0.07 0164072 015 317612 001650 430520 006600 494 456 080 060 02.14100 015.7800 00.556844 00668748 00.854942 00.751680 0172932 0768 0.824 0158378 015 187 608 015 060 538 060 recMoavsesred (gE) 00.00541396 00.00335833 00..00220980 00..0134297 018 00566789" 00..00907402 00..00650970 00.4018072 00438712 00430520 04% 00425160 00410 00..0187800 00..00558644 00..00668748 00..00654024 00..00576108 00..0172932 00..00786284 00.4065378 0187 0068038 600307 0 c 9.11.5 Summary and raw data; ppm Fin serum as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. This data, although supportive, in the opinion of the Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. 600308 Stata Que AREM:D6T32191-1163944.S1ERUM SAMPLES ADnaatleyostf:AnDalDysWis: 8/8/95 TThhee gsaasmp1lecsolalreectbeudrinne2d.i0nmtLheoDfo1h:1rmTaInSAaBtM9i5l0i-CQuwsaitnegr.0.T10hemLsaomfptlheessaerreumt.hen analyzed on a Skalar Segmented Flow Analyzer using the lon Specific Electrode (ISE) Method. TfsoIanmSpAsleBleebctuthfifevenergeiolsesacsdtdrtoehdrdeotuaognhedaatchhheesaratemefpdelrmeeinxacisengeitlcpearcitolrcboeedefedosirsetmhtehraeosupugorhteetdnht.eiasTlyhbseteetsmwi.geneaTnlhtiehse amplified and related to the fluoride concentration. Tphpemifnlsuotrriudmee,ntTwheasstcaanlidbarradtecdurivnethfeorrtahnegheisgohfr0a.n0g1e5w-a0s.1p5loptptemd aunsidn0g.t1h5e- 1.50 fSteanrdsardlsogaanrditshmamoppltieosn.weTrheetshteanndcaarldcucluartvede bfoyrtthheeSlkoawlarransgoeftiwsalrieneuars.ingAlthese curves, All resus below 0.0001 ppm appear on the raw data as #####. A quality control standard was analyzed every 10 samples to check for accuracy and drift. Raw data is taken from the appropriate calibrated range of the Skalar printout and Lsummiarin zed kon anaEnxyceslubssperqeuaednshtedeitl.utTiohne.sfinal results are adjusted for the 000309 fe 14 21 GROUP 1 Dose Leve:l 0 SUMMARY OF 6329-134 SERUM SAMPLES AMDT 111694.1 Solar Quod Sample I) Fluoride in Sample Fluoride in Sample Fluoride in Sample Fluoride in Sample Fluoride in Sample FlSuaomrpidleein FlSuaomrpidlee in Gpm) 2h Gem) abr Gem) Gp) Shr Shr (pm) hr p24bwr e48mhr F52548 079 046 043 017 044 226 088 GROUP2 Fs2549 052 055 040 057 039 091 083 Dose Level : 2 mg/kg GROUP 3 F52559 052 039 015 03s ND 087 076 DoseLevel : 20 mgkg GROUP 4 F52566 on 034 086 056 031 077 094 Dose Level : 200 mg/kg. GROUPS FS2567 062 051 065 066 042 089 on Dose Level : 1000 mg/kg 134S-RPTXLS 000310 Page 1 2 SUSMEMRAURMYOSAFMP63L2E9-S134 AMDT 1116941 proT mead) "m"a3n29l-1e3e4 Shot ute. BESTCOPY AVAILABLE BFaassssa oomo 2200 ooll 002% FFaasze oomo 2200 0000 o0ms Faz 0m 20 010 0 FFaassissds 00m2 2200 001100 00s4 EEaassseas oomm 2200 00100 00%% Fad 00 20 010 0s rFoossesse oomm 2200 0000 0064 FEaassees 000m4 2200. 001100 001%s Bee om 20 ol oss ooo oc Foss om 20 20 0o0n o0s m BFaasess oomm 2200 o01w0 003%s Eases om 20 al 0s Emassesl oomm 2200 0000 0ou% FBaassseila oNwb 2200 o00w 0N3D1 Eases om 20 00 0% BFoossssa o00ns 2200 o00w 023316 EEasseas oooo 2200 00100 0o%m Basa oo 20 00 0 oTossam. oooo 2200 00100 0os%s PPoasssseesss o00os 2200 0o1w0 005%% srs oo 20 ow. om 1ass.suMXLS 000311 Page 1 3 NORA SERUM CURVE 1 7-31-95 dic oIoo sLpEerY SCoTs D (ppm) (mL) Spiked (ppm) WmgoanomoomoumRon3 W we RuIwR swmmroemom n2RoWwme 8wEwR wmsaerramwow nomnuomm aEomRE emgmetmloomni R0RoOmE ERwo pmmmaasnonoowona2 moImoEamo por wp pao R3B0 OMomWE0I wmnoermmooonmonn30 Mm om R0ME an ASnE wl (vg) Recovered (og F) aw wom ww oem ew a mm ww om ww wm ww we ow ow - SC aom E el 1TT J1 TTT nen SERCRVINAVE 600312 semaencsurve 1 NORMAN Sfr E om ren SCod nn 0 wn mmpntowwn(opomm) oB3 ootmommwm (ml) Spiked (pow) (om wuFm)meo(enumF) ouom wSaebwsomaon gwpe wmm oer e soma o 3notmw onmm ofoomn doei oeumne Semmegson sgmm gmBoEen ommomooRnoooommwoaoaamnmowtwsrmooonwuonmmmm meowmoemgwor gomn Bgweoen ooont wn0Bosmm woommmo3 w@w mmsm ommwe seoTwEoERmon gouun smBmeasne wosmnenownoeommomomwomeee osgemoHeomm ououmn memegmoees w gsonm agBmuEre ooownn omn oowumm oommmmoeemnm oman unouRn swoemysomon gssy mgauieooomrnnoowmmm oomsmmanam moR emomRoTempEoeems o muen Tr eaam--ee fo | T T = T r T [| SERCRVIN.SUM 000313 Page1 b ans DOsii eptstoAnni pmowsCoausApLustA:tRsBeAeTnAUVDATASOTR8I1M PLot aWa 1 semen adme eos w iIe ewm om om DFofr mebi ToomES GEoTkp oh oSoTwm om wwoomwe 545T COPY AVAILABLE bSbooo SSAememsmi Boodmm ooonee 0e ooaooanm Gm owwmwm wm BRBooOoMaEmvmRen 0ihe je wiwmo omwe oosn nooaonn iw B [[I HS t nm ratdon 1 mia on B B Boh ha Bohn iIik% eComme om aaemow mo aooaosnm om Me om ooownw mmowww oomn aoxw n5Noo SaFmiaame 3 Ew aaa0 on BnHoooohon oodmmm dm note oR eg L5FoR FR Tha we iout ms oown H 0Bohmaw oo8ma em aw as ow ow 73 Bag 3BB o a waass 5 Sam ooruad Thi oo B BBoM hh iiih ootmmm amBmoe oconkw oaoenw imoowww {nSise noe aw 8 - Paget : ta onus PPoom w.8 P Pi`SiXE EEEEA REE BBEOOEFBBR PRB 1Pom2e~-zOB Pp:PToE5EmO.R0EoHR ow LBPPoamoWBE OOTEEET sEi5e F DTiHa 10 1s low Ea BfiBE fBTERmaEEaEIEmEmmamgoSmEEgoEEuouowEonrRweEgoyRmm i RE pou og BESTCOPY AVAILABLE EPTEROfEBmEosmmaBaEowaLnwoEsm FpumwEouEzoe ammoommoewomousow = ePsnite.etnsowtonuatohseDnATAMSosER LPPDo oootmsEeh aemmaoemoias moem om DSo Pooa oaswwoomm omn mw ooowm [BHB I o==no==oo iS i AILABLE bFE BE ooe mm oolm 5o>ommoowm 0 a em HSTCOPAYV BF FIo H Re iooaow n nnooo doooow 3F 3BmopA emaan &Fo5aS FnBBIooodmmeoSoowk Ny PF OoT A w hae owe o0finn om BBOoWmaOW em an oes em oom 2g2 & F PFoiy e)m FH 50o on nn nooo oeow ooamoen oomum aoomEm ow Mw o oos o muw i'< til o 2 BK oon 20 oo ie #21 n BOKE "id 2B0 owol 136 febie! ant Pv sans w5 olmoiom aoonan B nnooO woWoom ew am oon as om BDOiTmowe oe b1iiw om 3 ow ae om am on om ow bbaomamm oa@a BBnoodddeee iiman omtomm mmdeee ooolms ooonmw owwm PG$ ao mTE b05+ nBBoooaomok oom i Eomm M ote Won oowm bpboo oAme.n o5wwo nFnR oowmo F5FaR men boarnn Fnwr oomw en 5 3i mhmeeaa oorwnd FoR moom on 8 a55 rrmeTanhn ooooannn nn BBoooooo okk oooonom GQ a mwmow oiihe om wo a a bm m w toaeo nnBoooO ok Riinm am am oon a ou aa Twahom oe 2g & BESTCOPY AVAILABLE ~ Page2 anos 10 0a Shales WoT 729-1S3e4n Ansys Tle Nase GE\SKALATDATAVIORTA\SERIMIS505000 " 600318 1995-08-08 10:58 output of : Fluoride 1.5 PSiagtnhalnumtbyepre :: Da3bubbled DesycsotleonrNumber : 0Yes Reasitauntpele ii NNoo iaicl oTuhtrpeusthold i; 04095 Window event : Off s5i2 ssTTaannddaarrdd :: IIggnnoorree 5334 ssTTaannddaarrdd :: IIggnnoorree 5365 ssTtaannddaarrdd :: Ign0o.r1e50 5576 ssttaannddaarrdd :: 00..360000 50 sSTtaannddaradr:d: 11..250000 ODridmeernsi:onInv: erPsPeH Logarithm sStrairgtgerValLuienit :: 5108000 DSUec SoPteaaarkt sihigagnpnoeorree ::+ 1P62o00inteSSdeecc FiMelatseurre window :i 7H5o 3 fRoergaelnlearation : No output: aN.enk Fluoride L PSaitghnalnumtbyepre :: De0bubbled DescysotieonrNumber i: 0o alRtosuatmeple ii NToo aaiila oTuhtrpeusthold :; 04095 Window event : Off 950808A1 600319 1995-08-08 10:58 Output of : ss21 sstTaannddaarrdd :: 00..001350 ssi3 ssTtaannddaarrdd :: 00..006900 s$56 ssTTaannddaarrdd :: 00..112500 s7 s8 ssTTaannddaarrdd :: IIggnnoorree 55910 ssTTaannddaarrdd :: IIggnnoorree oDridmeernsi: on2 : PPM sttrairgtgerValLuiemit :: 5108000 DSeUc PsteAarktsihgnaorpee:: P6o0inteSdec MoeNadsureigwnionrdeow :: 71520 Sec FRielgteenreration :: WNoo foourmtuplaut:: c4d.:4z8c4d% 950808A1 06320 1995-08-08 10:58 Output of : 950808A1 Fluoride 1.5 Fluoride Pon on Pos Typ 1dent Ch Result 7 Time Ch Result F Time wiit ediw InitialTDrWraaicsfehtr 333 01l..e0463s56 230e6035 444000..l40i70r70e923 a00 iiS ow5512 sGttaannddaaarsra1dhz 333 00o.0l078i75 769203094 444000..0000290i72 53 Standara3 3 loss 1081 4 loki a00 SiS55i SSStttaaannndddaaarrraaas4s 333 000.ll2is02e59 1ai4ss3es3 444000l.l0115810%87 000 fiio13555857 SSStttaaannndadaaarrra89ad7 333 001..622173799 i12731853393 444 000...2344me6sd6 000 113 oaswlo standarDaWriafs1t0h 333 110.430s8205 222437802375 444 o000..04407s05i20s 000 1306 uoUu sSmeSMmPBBeIi2Kk-1z1 333 00011.318328 332018038842 444 000.l0.o2s31a 00 n33o04uu GSsmehxzosa6s02o22d 333 o00..l1217s33 3i353o306 444o00.l.0i294s57m00 00o 32i55u0ou SFRFKs3232ss344000322 333 O00r..02o73e64za04S2ee3a04s 444 000..l02033291877 000 5276 uaow F32r3ai5sf9he2 333 001...003783653 id4ssf2oass 444000.l00420s5i29s 00 2zEa 4u SFFss622%5626761322 333 000.020708840 55421s802486 444 000l.0o08d30i6si0 00 i33 uUou ssSekiakass60o2.2d4 333 000122427368 s5s4ee6is3n 44d00..o1214m4610 5s3% uuou BBBLiKixki2 333 0ool0ld01e%7 esei3e3es7i 44i00o..o0150u51z43 0 B9 dwou BBLWrKaiseh4 333 0i0.0s086ds5 6ce9es1sl7r 444o00..l00d0407s23 a0 0Q2ouuou sSPpB8Le2K2 333 000.11028263 77z033z838 444000ll0104o1d3s 0 3G5 uouuw sSSearasas 333 000z22447r9 777973i3e282 4440002..52262ms 0 9iuWou SR BBiLkK 333 000.102622 ee42o33s74r 4040.0.l12o17k3o06z o0 E5a0 oav 520u Ps2sBaersoi-htd 333 10o5087e14 s9dt6oa1ir3s 444o0l0ldoemans Fs2sio4 3 0.07 9138 40. a0 0 8a FSiss-4 3 0.073 930s 40.0193 Page 1 of 2 +0:06521 1995-08-08 10:58 Output of : 950808A1 Fluoride 1.5 Fluoride L Pru Pon Pos Typ Ident Ch Result Time Ch Result F Time 5545 uu FF5522556567--44 33 00..007752 99468684 4400..00126594 00 557 uu $F5522554498--66 33 00..007743 109081334 4400..00220116 00 5598 uu $SP5K255692--16 33 00..018012 1100139604 44 00..00377873 00 6601 uu SPSKPK25602--12 33 00..112148 1100751450 440.0.11100372 00 6632 wa DWriafsth 33 10..507615 1101818090 4400..40802032 00 665 uu SSPPKK 225500--32 33 00..139088 1111243195 4400..21682979 00 6676 uu P52566B-L6K 33 00..103803 1111756941 44 00..01413910 00 668 ua F52D5r6i7f-t6 33 10..507728 1121191450 4400..04382233 00 W7t0 wrw Runout WWaasshh 33 00..006655 1122355910 4400..00000022 00 Page 2 of 2 / 600322 Calibration curve of 95080801 : Fluoride L | 0.230173 [TEEICT 0 0otsaczMTTyea) c .3 ? { 8 I3 0 Order : 2 Measured r 0.90946 "00 60032; -- Calibration curve of 950808A1 : Fluoride 1.5 -- || 1.735084 [ETT WO00T 6a G0 ~ TYITHEA] J || | $ 4! 3 0.051501 0 Orde:r Inverse LogarithnMeasured 5: seu ass | Gocgzy Rau data of 95090801 : Fluoride 1.5 095 [YEE 5 WITH a0 [IPN ea] ". Lo + Ed Coe ` 2i # \ * a a . o A 0 0 oa Tine Pas | Lo 5000 Esc=Exit ! Fi=Help ! Crtl-P=Edit peaks ! 000925 Rau data of 95060881 : Fluoride 1.5 095 [ZIRE TT [PY 7594 I(EN ed) | i + 5 : 3 -Ey o>s od | 5 Lo = = 5:7) CAT 0 Fat) ow EsceExi~t~ | FizHelp | Crtl-P=Edi0 t pea0 ks | m Q * 2| =) gaia 07 000926 Rau data of S5080881 : Fluoride 1.5 Pathno: 3 Tine: KEN Ualue:SEETUNEDelta: [21 4035 w QQ : 3 3 6 @ El ee\ v TrA0en ve . o 8935 Tine Esc=Exit ! Fi=Help ! Crtl-P=Edit peaks ! || | 13935 000927 1995-08-08 14:27 output of : 95080851 Software : version 6.1 c1990,93 operator + ow Date of the Analysis : 1995-08-08 10:58 Analysis File Name C:\SKALAR\DATA\EWIDATA\SERUM\350808B1 cou hsjas [RS wWomn aLCY ont CFalluiobrriadteio1n.5order = Inverse Logarithm s1ope ry Result = 100 [x=-=el] xci s == = ccoorrrreecctteedd vvaalluues ooff tthhee Slope of the electrode csaomnpcleentration 1 aa12== -00..0000006090 20 = -121965 CFalluiobrriadteioLn order = 2 Correlation : r = 0.99948 Res=ua2l*tx? +a* xl +a0 aazi== 00.0000002030 20 = oloosss sampler HTuympbeer i+ i5A1000 WSaasmphle TTiimnee :: 15200 sseecc.. MTarke upTime :: 1Singlseec. snpeeecdilael Height : N7o0nemn. Diluter dnieleudtleionHeiFgahcttor :: 8100 mm Rdeislauntpiloen Volume i: a2.5 ml. Dilution runs :1 URseeprrocfeisle :: No Lome 600328 : 1995-08-08 14:27 output of : Fluoride 1.5 SPiagtnhalnumtbyepre :: De3bubbled DesycsotleonrNumber : 0Yes aiRetsuatmeple ii oHo 44i51G Tohuretsphoultd:: 40095 Window event ; Off 5521 ssTtaannddaarrdd :: IIggnnoorree 5534 ssttaannddaarrdd :: IIggnnoorree 655 ssttaannddaarrdd :: Igno0r.e150 5578 sstfaannddaarrdd :: 00..830000 9510 ssrtaannddaarrdd :: 11..250000 ODridnearn"s:ionInv: erFsPeN Logarithm sttrairgtgerValLuienit | 1580000 DSUec Poa shape: Pointed HoSotaaAsrutreiiggwnnioonrrdeeow :: 7615020 3SSeecc RFeigletenreration i| Noo ofourtnpiaut: 44.808 Fluoride I PSaitghnalnutmyepre :: De0bubbled DescysotleomrNumber i: M0o aRiestaumtpele iNoo aaiila oTuhtrpeusthold i 04095 Window event ; Off 95080881 600329 7 1995-08-08 14:27 Output of : 95080881 sl sTandard : 0.015 s2 sTandard : 0.030 8s34 ssTTaannddaarrdd :: 00..009600 s$56 ssTTaannddaarrdd :: 00..112500 $s78 ssTTaannddaarrdd :: IIggnnoorree 58910 ssTTaannddaarrdd :: IIggnnoorree order : 2 DstiamretnsiVoanlue: PPM : 500 DU PteraikggesrhapLeimit :: P1o8i00nteSdec esNtdArt iiggnnoorree :: 16200 SSeecc FMielatseurre window :: N7o5 % RfoergmeUnlearatio: nc4:=:c3No output: #.448F 600330 : 1995-08-08 14:27 OutPut of : 95080881 Pluoride 1.5 Fluoride L Pry PrH Pos Typ Ident Ch Result F Time Ch Result Time w1 t iw InitialTraWcaesrh 33 01..046608 21605 44 00..70402504 00 23 oaw DWraisfht 33 10..046800 368268 4400..07045646 00 45 ss12 SstSatnanddaarrda 12 33 00..007614 971317 44 00..00219475 00 76 5s34 SSttaannddaradr4d3 33 00..018097 11028651 44 00..00692014 00 89 ss56 SSttaannddaarrdd 65 33 00..112565 11641337 44 00..11511722 00 1101 ss78 SSttaanndadradr8d7 33 00..621861 11798671 440.0.42350777 00 1132 ss910 StSatnadnadradrd109 33 11..426380 22311317 44 00..76451373 00 1151 wa DWraisfht 33 10..046807 22478279 440.0.07045946 00 1176 uu 5$522555499--88 33 00..006761 32081400 4400..00127843 00 189 uu 552255667--88 33 00..007731 33138682 4400..00238218 00 2210 uu FP5522554489--1122 33 00..006678 33751324 440.0.00129250 00 2223 uu FF5522555696--1122 33 00..006645 34086738 440.0.00115364 00 2254 uu FS5P2K56762--112 33 00..101627 44421335 4400..00927078 00 227 wa DWriafsth 33 01..046800 44852849 4400..70406584 00 2289 uu SPSKPK25602--12 33 00..133324 45913193 4400..12294179 00 3310 ouu SFPK52524580--82 33 00..037962 55425899 4400..03328573 00 3332 uu BBLLKK 33 00..009952 55861442 4400..00678126 00 334 uwu BBLLKK 33 00..007768 56919604 4400..00431999 00 3376 uu SPK 62B-L1K 33 00..009787 66531440 440.0.00747102 00 3398 wa DWraisfht 33 10..046809 66695209 44 00..70500534 00 0auu SPSKPK25602--12 33 00..121194 77201460 4400..21007821 00 4432 oouu SSPPKK 225500--32 33 00..223046 77359605 440.0.12929371 00 4454 oouu SSPPKK 225500--54 33 00..225833 77971451 4400..22538797 00 4475 ouu BBLLKK 33 00..016510 88028491 44 00..10405616 00 i49suou FF5522554489--2244 33 00..102830 88463190 440.0.01415279 00 5501 aw DWraisfht 33 01..048608 89071858 4400..07043948 00 5523 uu PF5s2s5s6s6--a2d4 33 00l.0077y6 o93d1s0 4400..0034837 00 Page 1 of 2 600331 1995-08-08 14:27 output of : 95080881 Fluoride 1.5 Fluoride L Pry Pry Pos Typ Ident Ch Result F Time Ch Result F Time 5545 uu FF5522554687--4284 33 00..007799 99468672 4400..00444369 00 557 uu FF5522555499--4488 33 00..007757 109081337 44 00..00431778 00 5589 uu FF5522556667--4488 33 00..008704 1100138671 440.0.00345781 00 6610 uu BBLLKK 33 00..009895 1100573096 44 00..00758684 00 6623 w4 DWraisfht 33 10..046807 1110182876 4400..70409564 00 6654 uu BBLLKK 33 00..008888 1111243162 44 00..00661047 00 6676 uu SPK 6B2-L1K 33 00..018141 1111578660 4400..00956332 00 6698 dwa DWraisfht 33 10..046802 1112913656 4400..07045745 00 Wt rw RunOut Wash 3 0.060 12410 4 0.0054 0 Page 2 of 2 00332 |i| 1 veumar Calibration cure of SSORCER! Fluoride 1.5 ---- Channel 3 Measured:II Concentration: [INGER Delta:[ER | 7|1 | |j | ii | :i | ' || | ||i pp" : Orie rman Loeriei [ si seu ws | 000932a Calibration curve of 95080881 : Fluoride L 0.2096763 Channel 0 Heasured: SH Concraetinont: IVR} Delta:(ER Pp : 5: d4g 0.005427 2 Order : 2 Measured r: 0.99948 500 | 606333 ou dats of 5500081 Fluoride 15 ---- || w= YTTHEERITTE 0 WY 77[nrc | |~ ow 4 ~ i!i . o o . A <4 we PY Oa E00 aan Po 0 Toe som Esc=Exit ! Fi=Help ! Crtl-P=Edit peaks | 600334 "085 I Rau data of 95080881 Fluoride 1.5 | + & | $ :i oH a 1I | Pb4 . 1" hy A ay 8 i i | 3717 aon | oth] 4560 Tine ' Esctxit | Fishes | Gril FoEait peaks | irae | 9560 000935 ; fou data of sn0t Flori 5 |i 035 IEEE 695; WANTS see D(Edca] | Ww @ 2ii | wp ide gpfaial | GeU1 a 0 os = Boome T FiTieln CHTPEAI pemks rl] 006336