Document 2q26rGBBnyxdpXVk4OrBqoX0N

PR226-0225 3M Environmental Laboratory Final Report- Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T-6051 and T-6054 in Rabbits In-Vivo Study Reference Number: HWI#6329-133 TSetsutdySuNbusmtbanecre:: AFMCD-T12-901(3T1-96055.11 and T-6054) `Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facility: 3M Environmental Technology& Services 935 Bush Avenue St. Paul, MN 55106 Method Numbers AMDT-M-1-0, and Revisions: Thermal Extraction of Fluoride by Means ofa Modified AMDT-M-2-0, Dohrmann DX2000 Organic Halide Fluoride Measurement by Means of Analyzer-Liver an Orion EA940 Expandable Ton Analyzer AMDT-M-4-0, AMDT-M-5-0, Extraction of Fluorochemicals from Analysis of Rabbit Liver Extract for Rabbit Liver Fluorochemicals Using AMDT-M-8-0, Electrospray Mass Spectrometry Analysis of Fluoride Using the `Skalar Segmented Flow Analyzer with Ion Selective Electrode Initiation Date: See attached protocol Author: James D. Johnson Approved By: ames D. Joton Study Di r ez/2s Completion Date C00269 fSaabmrpicle(sT-o6f05li1v)erwefrreomanraablbyiztesdaadtmi2n8isdtaeyrsedpoFstC-d1e2r9ma(lT-ad6m0i5n4i)storraFtiCo-n1f2o9r treated total loirqguaindicfofrlmuuolraitnieoannddopseerfglruoourpoo(c1t2a.n8esmugl/fkonga)tet.herTehies results on the show that for the highest average about 0.2%of the dose in whole liver at 28 days. a`Tbhsuosr,ptdieornm.al administration of FC-129 at higher levels results in some dermal Two 138) studies were performed on and this dermal absorption FC-129. A study (HWI pharmacokinetic study (HWI#6329#6329-133). The pharmacokinetic study showed absorption of that perfluorooctanesulfonate is a FC-129. Liver, serum, and other useful tissues `marker to assess the dermal were available for analysis by acnoamlbyussistioofnspfeocritfoitcalmoolregcaunilcesflsuuocrhinaes apnedrfellueocrtoroocstparnaeysumlafsonsatsep,ectBryomoebttrayinfionrg and atshseensasnmaelnytzionfgthdeateaxtfernotmorfabdbeirtmsaalta2b8sodrapytsiopnosotf dFeCr-m1a2l9diossep,roivnifdoerdmaitnitohnisfosrtutdhye. 3.1 Test, Control, and Reference Substances and Matrices e3.q1u.i1vaAlennatl,ytical Reference Substance: FC-95, lot 161 or 171. They are 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 g3.r2ocSeoruyrscteoreo,f bMoavtienreiaslesr:u3m MfrIoCmPS/iPgCmPaDCihveismiiocnalfoCroFmCp-a9n5,y.bovine liver from 3.3. Purity and Strength of Reference Substance: Responsibility of Sponsor. 3.4 Stabilityof Reference Substance: To be determined by Sponsor. 00270 2 b3i.o5loSgtiocraalgseaCmoplnedsittihoensstofroargTeeisst-2M0a+te1r0iaCl.s: Room temperature for FC-95. For G3.L6PDiRsepgousliattiioonn offorSptehecitmiemnesp:erBiioodlroegqiuciarletdisfsouresstaundidesflluoindsgewriltlhabne retained 28 days. per 4.0 EXPERIMENTAL - Overview `The tissues analysis for ffrluoomrianneimcaolmspoduonsdesd.as Adtestchreidbiesdcr(eHtWiIo#n6o3f2t9h-e1S3t3u)d,ywDeirreecatvoari,laablseerifeosr of canoamlbyutsictailotneswtasscothueldmobsetpelirkfeolrymaenda.lyTsihsetsocprreeesneinntgdfeofrinfiltuiovreiddeatian floirvearbsvoiraption. cOhtrhoemraatvoagilraabplheyt/emstassswesrpeecetlreocmtertosrpyrfaoyrmmaestsabsoplietcetsr.osLciovpeyrasnadmgpalses were analyzed by dbeorthmaclomabbsuosrtpitoionna.nd electrospray. Data were then analyzed to assess the extent of D5.o1hrAmMaDnTn-MD-X12-000,0 Thermal Organic HEaxltiradcetAinonaloyfzeFrl-uoLriivdeer by Means ofa Modified E5.x2paAnMdDaTbl-eM-Io2n-A0n,alFylzueorride Measurement by Means of an Orion EA940 5.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver E5.l4ecAtrMoDsTpr-aMy-M5a-s0s,SApneacltyrsoimsetorfyRabbit Liver Extract for Fluorochemicals Using w5.i5thAIMoDnTS-eMle-c8t-iv0e,ElAencatlryosdies of Fluoride Using the Skalar Segmented Flow Analyzer 6.0 DATA ANALYSIS The data are attached. The leveloftotal organic fluorine in whole liver for the ttchhoienstvrmoaellt,uhe0os.d1a.2r8e,Jous1nt.2tu8h,seiaonnrgddmeefratoberfric1r6edaoudsgie/nswghsaoraelnebdeleilvxoetrwr.atphToelhaeptria1nc2gt.i8fcarmlogmq/uktahgnetdistetaratmniadolanrddloicmsuietrvfeo,r however, results in detectable amounts. oforganic fluoride in whole liver at 28days 00271 $ tphoest6droasbeb.itsTh(eF5v2a8l9u5e)s rtoan7g5e ufgr/owmhobleelolwivtehrefporrarcatbibciatl Fqu5a2n8t8i9t.atiTohneliamvietrfaogreofnoe of 45 ug/whole liver with a standard deviationof 21 ug. Electrospray mass spectrometry data are in agreement with the combustion data; hgt1hr2oe.owr8ueepvm.eigsr/,Svkmetgarhlyedlsoleisdteetaltregeercptoeeaursbpftl,lieumpoaaertmroefoodlucuttnooatrnbsoeeosacurotlenafnootebnhsaseuteloerfrvoidennedartthieonefitgs1hr7edoeuu1tp.ges2/c8twoehtmdohglei/renktaLglhilvaaenrnradtbohbrfeiatlb1er2Lsisi.c,v8egrm`rgFo/uakprgee 72s55amtupogl/e8ws5haoutlge2/8wlhidovaelyresolfpiovtseortta(dlemorermaganalonifdco4sfe8l.uuogrTi/hnweehoaflomreouclnoivtmesbr)ua.srteTiheosnetairnmaaabltbyeisditsthoa(trFa5sn2hg8oe8w9fe)rd,omhad 56 ug/whole liver perfluorooctanesulfonate. From the pharmacokinetic study on FC-129 (HWI#6329-138), it is known that a aipgnpeorporfeadlaburbpoioritronstoilciiostvna>enr1oefasmtutolh2nfe8otnihdan.attyersT.ahvauesIsntp,oieusrsiffkdlFnuoCoos-wre1onow2ic9tlthlaaintsbeedtsheuberlifmhooaatnllrafla-tylneis,affbeosorofmrebpdeerdtfoalupoorrotoicotnanoefsiutlfwoinlate prFeaibrfbftiyltuusograosposecrutfmalinuenosgruolaofbcotonaadntyeesweueqliufgiohvntaaltoeen/f2twshko(glFeCa-nl9id5v)eer.xpisrAef0st.se2ir5n%agonfthienthtderoasdveoesnieonufpsoordtoatshseesio1u2fm.182.m8g/kg dwmaigyt/hk2ag8.baiTorhlauobgbsii,tcaehlsathdiamla1f-t.le0id5fe%loefovefl3ts0headfadeyorsseawnoiniunlltidrvaebvreeanatop4ups8rodhxooiusmreast.oeflF1yo2r.06.c5mo%mepyoakfrgitshwoeonud,losd1e.b0aet5% 20182..5d8%amyosgf./tkhge wdoosueldinbewh0o.l2e%loivfetrhaenddoessetiinmawtheodlleevleilvserafitfetrhedelremvaellsaadrmeinciosmtpraatrieodn aotf SeOltteuhcdetyrroDddiaerte(acstewoearsaipscpoenlnoldteiccrteeesqdu)i.ursTeidhnigtsoSdkraaetlaaac,rhastlehteghmocueognnhtcelsuduspfiplooonrwtsitavaneta,eldyinzheetrrheewioatpnhidniitoohnenroseeffloetrcheteiivse not discussed in detail fp6l.ru1oobrCliinearmtcewudimtcshotmtahpniocseuasnndatlshyasitinsMtihaseytlhiaHvteartvhaeet e4Ax8ftfehenoctutroesfd bitnihotethreQaunpashlfaiortrmmyaacotofikoitnhneeotfDitachtesat:ruesdTtyhoef the mc(ooHnlWseIic#due6lr3ea2sb9lo-et1hb3eir8o)ttrhtaaonnsppfeeorrrffmllauutooirrooonooocfctttaanhnceessuuslleffvoeonrnaaatlteepiersrenpcoretenstokennfto.wdnoHs.oewTethhveaetrref,lcutoohurelind1a.bt0eed5%of the mdoasrekeorbsteoravsesdesast b4i8othroaunrssfostrimllatiinodnicoafttehsitshactopmeprofulnuodr.oocAttan2e8sudlafyosn,attheeivsaalassenssiotuilvde 000272 (mbseeitnsacobeomiletiwisshma.mteaIhsfitughrhieesrdwtaehtra2en8ttrdhueaey,asb)tohaevebvuaeilslutteiimnfaotcreodmoepfrem0na.sla5t%aiboosfnortfphoetridtoohnissheaddseulbeayytodaendfdeinltiahtyeieovdnalue o0ffd0o.s2e%forfotmhethdeosinetarfatveerndouesrmdaolseadtmhiantiisstrtaotoiloonwi.s being compared with a percentage a`Tdhmeirneisitsreavtiidoennocfeaof12d.e8rmmagl/kagbsdoorspet.ion in rabbits of FC-129 after dermal 8.0 MAINTENANCE OF RAW DATA AND RECORDS a8.p1proRpariwatDeastpaecainmdenDs,ataan:d eRlaecwtrdoantiac, daaptparwoivleldbpermotaoicnotla,inaepdprionvtehde fAinMalDrTeport, archives. 9.1 Protocol and Amendments A9b.s1o.r1pPtriootnocToolxiacnidtyFiSnatludRyepoofrtT:-6H0W5I4#6a3n2d9T--163035:1 "iSniRnagblbei-tDso"se(PDreortomcaoll type TP3016.AB for dosing of animals, tissue collection, etc.) 9.1.2 Analytical protocol AMDT-013195.1 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: See methods 9.8 Quality Control Practices: See methods 00273 5 9.9 Test Methods: See Protocol AMDT-013195.1 9.10 Instrument Settings: See methods 9.11 Data: See attached. e9x.t1r1a.c1tiSounmfmoallroyweadndbyraawnadlaytsai;s ug F" using iOnriwohnolieonlaivnearlyazserd.etermined by thermal 9.11.2 Summary and raw data; analysisofliver extracts using electrospray mass spectrometry. e9x.t1r1a.c3tiSounmfmoallroyweadndbyraawnadlaytsai;s ug F" using iSnkawlhaorlseeglimveenrtaesddfeltoewrmainnaeldyzbeyr thermal with ion selective electrode. 00274 6 ---- 9.1.1 Protocol and Final Report: HWI#6329-133: "Single-Dose Dermal Absorption/Toxicity Study of T-6054 and T-6051 in Rabbits" (Protocol type TP3016.AB for dosing of animals, tissuecollection, etc.) 0027s5 2? LPWOSIET SEOFCFRIOCEFNBAOSXRI7A5N4H5 Sponsor: St. Paul,3mMinnesota FINAL REPORT Study Title: SSitnugdlye-oDfosTe-6D0e5r4maalndAbTs-o6r0p5t1ioinn/TRoaxbibciittsy + CORNING Corny AEST {JUN 21 ss C2 S Author: Steven M. Glaza Study Completion Date: June 16, 1995 PerformingLaboratory: Ha33z0l1etoKninsWmiasnconBsoiunl,evaIrncd. Madison, Wisconsin 53704 aboratory Project Identification: HN 6329-133 Page 1 of 46 EPXrPoRnEeSS6M0A3I.L241D.E4L4I7V1ERY 3301 xiNSMAN BLvD 00276 MADIFSaOxN. 80w8r.241.s7a2ie2s7 Page 2 of 45 HHI 6329-133 QUALITY ASSURANCE STATEMENT WT{iiTsOtRcs)onrsSeiopnoo,drtLIanhcba.so,rabteioenrnyacrPcerovaricdetawineccdeebRwyietgthuhleatthQieuoanlFsio.otyy AasysurDraungceAdUmniintisotfraHtaizolneton sSSTttfhaueandndyfaaonrDlddilironewgcistnograrieannsdipsemscautenidaognestmoewneHtra.ezlectWoornnidtumtcaetnneadgsetsmnaedtnu2ty}fimncprodeagi,stnongoss]g'y. ArdCehpio(nrbt)sepde(6c)ttoi(o7tn)hs.e operating procedures. according to Inspection Dates --fron To 1122//2088//9944 1122//2089//9944 0011//3009//9955 0011//3009//9355 0033//1177//9955 0033//2211//9955 0066//1164//9955 0066//1165//9955 ----Phase RepoDratteed to Date to DPorsoetocAodlminRiesvtireawtion study Director Management 1122//2089//9944 01/10/95 PPrroottooccooll AAnmeennddnmeenntt Data/Report Review 0011//3009//9955 03/21/95 0102//1100//9955 02/10/95 DRaetpaortReRveireewview Report Rereview 0063//1251//9955 0044//1100//9955 07/10/95 06/16/95 07/10/95 J) RCeepcrielsieantMa.tiDvaeg,arQuality Assurance Unit fate C-1 -75 C0027 Page 3 of 46 STUDY IDENTIFICATION SSitnugdlye-oDfosTe-6D0e5r4maalndAbTs-o6r0p5t1ioinn/TRoaxbibciittsy HHI 6329-133 Test Materials Sponsor Sponsor's Representative Study Director Study Location StSutduydyTimIentiatbilaetion Date IExnp-elriifmeenEtnadl Da(tIen-Tife) Start Date SEtxupdeyrimCeonmtpalletiToenrmDiantaetion Date 21.. 7T1--66005514 M ToMxeidcioclaolgyDeSpearrvtimceent P3.M0.CenBtoexr,332B2l0dg. 220-2-02 St. Paul, MN 55133-3220 3John L. Butenhoff, Pho ToMxeidcioclaolgyDeSpearrvtimceent P3.M0.CenBtoexr,332B2ld0g. 220-2E-02 S(t6.12)Pau7l3,3-M1N962 55133-3220 HSatzelveentonM. WiGslcaoznasin, Inc. MPa.0d.isoBno,x W7I54553707-7545 (608) 241-7292 Hazleton Building NWoi.sco3nsin, Inc. M3a8d0i2soPna,ckWeIrs A5v3e70n4ue DDeecceemmbbeerr 13, 23, 1994 1994 JJuanneuar1y6, 2159,915995 June 16, 1995 00278 Acute Toxicology SSttuedvyenDiMr.ecGtloar.za Manager SFtruadnycisCoo(rBduid)natW.orMcDonald IPna-t1riifceiaSuPpaedrgvhiasmor RReopseortM. SuBpreirdvgiesor Quality Assurance MSahneargreyrR. W. Petsel Page 4 of 46 KEY PERSONNEL HT 6329-133 Laboratory Animal Medicine DCiipnldoymaJt.e,CarAyC,LAMDVM Supervisor Anatomical Pathology ATnhaotmoamsicEa.l PaPlamtehro,iogPihsDt DJeabcokraSherLf.ortP/irkel NSeucpreorpvsiysors ASnunpeervMiossohrer Pathology Data 00279 Page 5 of 46 CONTENTS SQtuuadlyityTdAesnstuirfaicnacteioSntatement SuKmemyaPreyrsonnel ObRjegeucltaitvoery Compliance TTeesstt SaynsdteCmontrol Materials RPersocueldtusres SDiigsncaustsuiroen RePfaethroelnocgey Report Table 21 IInnddiivviidduuaall CalnidniMceaaln SBiogdnysWeights (g) 43 IInnddiivviidduuaall DPeartmhaollogIyrrCiotmamteinotnsScores 5 Individual Aninal Tissue Weights and Bile Volunes AppPernodtoicxolA Deviation PPrroottooccooll ATnPe3n0d1n6e.n4t8 No. 1 Protocol Amendment No. 2 HAL 6329-133 Page 23 36 88 89 1130 113 11s4 1168 220 27 320 a312 45 00280 Page 6 of 45 HHI 6329-133 SUMMARY sTkhiins sitrurdiytawnacsy dofonTe-6t0o54assaensdsT-t6h0e51sywshteenmicappalbiseodrpttoionth/etoxshiicnitoyfraandbrbeiltast.ive oTfhethsetudHyraw:a(sNZc)oSnPdFuctsetdraiunsinfgortheraeceh mtarleeatmaenndttghrroeuep.female acclimatedrabbits --Group TestMaterial Dose Level _(ma/ka) Number of Animals Males Females 1 (Control) 2 StTe-ri6l0e54water 0of.128 3 T-6054 1.28 33 3 33 3 4 5 T-6054 T-6051 12.8 b 33 33 b& mAaAdtdmneiirnniiiassltteerr(eefddabraaitsc)aa. 1do0s-ecmvxolu1m0e-cmofse2c.t0iomnl/kogf. test SoToifheptphbceatcirkvaebobfimtaesta.ecrhiaTrlhaebbatittretahwetasmeincntldiispcipatetedesd frdreoemseaeionlfeedvhealiirntwaaacsntd.adamTishnieinsgstleepreeddoosfetaopoptflhetheckin ddrveeeasrrsneirpnavgpepreefdodrwwiiatthh24-SahaorguaarnuzWeexrpabopasnudraaegnedpeEsrleiacosudtr.oepdlwaistth tpaappeertotapperovairdoeundan31o1c cicuasgiejcssaetsinogn dgfoplbrhisoneui2r8rcvsaaltiaofontbsesreranvtdaestttiwoinocsre cwoea nrtderaoyclomnomdrauttceatrleiidatlyprcaehddemocisknesisawtnredarteaitocnoa.npdpurcAotdxeiddmiatdtoaenilalyly 1,toho2an.m5ie,trarland bpaeunfrpoporoiesfeestd,eatsyetsbr.emfionraBetoidoytnewste(iDgoahyrts2c9o)nv.terroelThdemeatteiernrniitiainlaeld addoenmrimDnaailystri-a9rtriifotonartir(oDananydre1os)mtiinaagnsngetw.eart er made 3y[e05ardsmiinncguo)t.l,elsectSaeufodbtrseercqounbetarnnotdlagreemaatdreiermnoigvasallofad(dDmeairynmia2sl)traaitnrdiroiontnat(iDroaenycsorwde4erdeandmaas8d.ethea"pBpDrureyoe'sxsiammaptleelsy . oaifnndibtl2a2oa.otdiownIans(aDodabdytifatr1ii)oo,mn,aappamtarrogxtihinemaalttienleeyaro2f4v-eihnnoeucrrosofpsptyhoesotndaonDsiaemyal(s2d8a,ybe2fa)op,rpsroonxiinnD.aa]ytisegely4,'208,'nt15, S2jeperppnraiornxaaittneiadotneInot1fo-cntsheexrnuem1id-nca-mnfidrfsoeneccetlpeihlaaucshlearanainfdmraaclnt.eicornosAp.lsliesdAa.lmlplTeahsneimwwaehlroselewcoenrlteirvieerua,tghebadinliaza:ecddanat aonlinmealcst.ed(2antd bnoetchropksiydneaynsd wferiosomhneodnoef(mtvaholeleumdeearnmdoanlloyneadpepftlaeimracmlaietnieoidnn Dood samples (serum and cellular ssaicthe gfrrooump awlyere for bil) "gnc Siant-eTsi,feanpdhaskei.dneys were sent frozenfrtaoctitohnes)S,ponlsiovrersa,ftebrilet,ermdiernmaat]ioanppolF ctaotion 00281 Page 7 of 46 cAhpapnlgiecsatiionn boodfyTw6e0i5g4ht agndainT-o6r051macdriodscnoopticresfuilntdinigns anaty HHI 6329-133 nteecsrto material-related HfiePmmpbaeslaerethdaenicmTlaailsntitc2ra2elaldtyaeydnsowrimoatflhstTtu-hd6ry0o.5u4ghToahtuits1.t2ah8neimmsagtl/ukdywyaswtihtaahtlsocthhneiontegexiccpehspyt.iwoenakAelolnfedaanrhiimnadls pdfauetrceiersnigaolnt-hreDealsyaatm8e.pdl.e TchToehlseleeccfotinintodrnionlpgrsomacaterederuircaeolsnsaainndddertaeerdsettonmotabteecrodnusidteerseadostionhjaupvrriytn"geisnstmcaulrlred dopefrrom1da-ul6c0e5i4rarniyattadteairomdanolseiniTrferivievtleatioaonfni.m0a.l1s2N8oatodretrhm1ea.l2812i.mr8ar/ikmtgga./tkion1-w0ia8as4l obp1s.re6er0tv5ee1ndddinveger|Lyroyessluilgtht g dose loys 00282 Page 8 of 46 The objective relative skin of this study was irritancy of test OBJECTIVE to assess the systemic materials when applied HHI 6329-133 toxicity/absorption and to the skin of rabbits. REGULATORY COMPLIANCE ATdhmisinisstturdaytiwoans'sconGodoudcteLdaboinrataocrcyordParnaccetiwciethRegtuhleatUi.So.ns Food and Drug Laboratory Studies, 21 CFR material mixtures prepared 58, for with the exception the Groups for Nonclinical that analysis of the test coornicgeinntarlattieosnt, mahtoemroigaelneiutsya/gseol1uobgilciatny,notanbde2,stlao3,bciatlaeindtdya4wlatashnoiumngaoltscofnodructed and the retained in the compliance with study file. All procedures the Animal Welfare used in this ha study copy is are in Sponsor and study previous work. director, the Act Regulations. In the opinion of the study did not unnecessarily duplicate any dentification TEST AND CONTROL MATERIALS The test materials were identified and described as follows: T-6054 T-6051 Amber liquid White plastic sheets aLThacebloercaaortn,otrriocelosl,omraltLeeosrtsiaNloT.iqwua8is6d-.7S4t8e-rDiMl-e02W;ateErxp.foMrarIcnhjec1,tio1n9,9)U,SP an(dAbwbaosttdescribedas Purity and Stability deTtheerSmpionnastoironasssu(miensclurdeisnpgonsuinbdielrittyestfocrontdeisttiomnast)e.rial purity and stability comantceernitarlatmiioxnt,urehsomopgreenpeairtedy/sfoolrubtihleitGyr,oups 2, 3, Analysis of thetest and 4 animals for wreerqeuecstoendsidbeyretdhetoSpobnesoard.equaTthee pfourritthye apnaudnrdpsotssatebasiblioilftiyttyhoifwsasthsetnuodtcyo.nctornodlucmtaetderoiral Storage and Retention The test materials were stored refrigerated. A stored at room temperature. reserve sample of each test 00283 The control material was and control material was Page 9 of 45 L3kenfoarnd 1w0ilylearbse Qperating rinetaaicnceodrdainncea wfirtehezeHrazlseettontoWmisacionntsaiinn HHI 6329-133 a(Hitle)mpoerroartaurrcey of-20c dSfiepsmocanaisrnodirendgafcatPoferntrotecrrceoodlimusprmsleauett(eiSroOinPa)l.ofisAanllyretuianniu-nsleeiddfetfoeprshtasoemtahteaercrciotareldstiwinangsgtoarnedMtuIwrinleuydp.msooAt inbhyre ance of the final report. Safety Precautions Tanhde ptoelsitciaensd. control material handling procedures were according to HHI Sops Test Aninal TEST SYSTEM fKfaallciaitmlaiztaoylob,aitnoMI3,8r0a2bonbPiaDtceskceeormfsbetArhvee1n4Hu,rea,:1(9M9Na4ZdWi)asSnoPdnF,masWitinrstacaioninnsewidenr.eatprthoecurHaezdleftroonm HHRoPt,onIrnscs., Housing cpDhouofnruttisernerogdlreainccecilspictmr,aeteinto-hnbeotaatnnodimmatlhsstraowiuenglrheeosustacctsthleeienlasttuecddayg,efsotrhiena apneirmiaolds owferaet temperature: iTenadsitvid7uadlalysy. andhumidity- Tt1h38ei-sphetoauircnolneTdd3iigthqtiuteoa/mr1pt2ee-rrhasot.uurredEaonrfvkir1o9lnimgehtntotian2lg3Ccc,yocnltaer.orleslaItnfiovcreastehhseumwiahdneiirtmaeyl ovfarroio5am0t%iwse2er0se%.fsortoa'nndfo a adverse effectnsonextihsetesdt,udythoeuytcwoemree. documented and considered to have hag mo Animal Diet [LhaeboraantionraylsRawbebrietpDrioevtidHeFd #a5c3c2e6s,s Caonnatlaymzied by the manufacturer for nPtuHotIrwiFatetieeodrnsa,aldIcnIcoi.mbpiotnTuehnnetasnfdeeadnad missenaysriuarrccetndimneaanlmtyoaulnt of wienrteerfneorneakdnntosww.inthcoSonarmtpaalmfeifsneacnottfesdthintehewtaherteesrfueleatdrseoropfewraitthoeedricstaautldlyyl.evaenlaslyztehdatbwyouHlHdI. havTehere Selection of Test Animals wpTrehoregeraapnnli.amcaeldTshewienrtreoandsiotdmueidnzytaitgfiriooenudpsbbyoduysainnwigneailgahtsnsutmrwbaeetrriefiadendedtebcroomdriyrneewsdepiognohndtinDgaryaena9dr.omtasg"ehsoantn.nd C0084 Page 10 of 46 YSeoiudayhtdivdarinaottioenxceoefd th2e santiannadlasrd fdoervieaatcihongsrouopf HHI 6329-133 tohfeemaecahnsweexigshetl.ected for the Stbrhoeaddyy5w%peripigrohortbsatboiflotirrteyeaatclmheevnegltr.oduupe`oontfeo efpaeocmoharlesheeaxanliwtmehar.le another fe n(oNot. stFa5t2i8s0t)icwaalslyredpisjpnaedceertqnhctinat'rtcehne This aninal was replaces male (No. F52877). mtr Study Design Ainnitmoaltshewefioglhlionwgingfrostnud2y,15gr7outpos:2,508 g at initiation of treatment were placed Group Test Material Do_(snea/kLae)vel NMuamlbeesr ~of FAenmiamiaslss 1 (Co2ntrol) StTe-ri6l0e54water 00.128 33 3 T-6054 1.28 3 33 3 45 T-T-66005514 12b.8 33 33 ba AAmaddtmmeiirnniiiassltteerr(eefddabraaistc)aa. 1do0s-ecmvxolu1m0e-cmofse2.c0tiomnl/kogf. test Justification for Species Selection cHhiositcoericbaelclayu,sethoef tNheew lZaeragleandamoWuhnitteofalbbiancokgrroaubnbditinhafsormbaeteinontheonantihmiasl spoefcies. Preparation of Exposure Area PROCEDURES JoOnhb,etatichnleipudpnaebydlebmeaifrseohareedmatdesesktinu)pm,ataeptrhpieraolxfilmaaanpktpselliycoaft2i0eo%anc,hofrtathehbebibtatoctwkaalsandcb,oldiypipfesdunrefcfaercseeesarwoyrFenN(:otior. aJInhriermiattlaestsitownes,rieteocsrlid(peipfneetdcatcston stDkhaianyt)sw8owuelradendpi2rn9escptleoucdteeadidthfeoinrusvieinstuoeafrlfieaznriyinnggoftlhatesheaopnapsnl.imjaelast.ionThe sites. Dose Administration mAater2inailm.alsThreecdeaiyvedofatrseiantgmleentadwmaisnidsetsriagtniaotnedofastheDayres1pective test or control CO028s Page 11 of 45 {Snryoeuapch1. ( aniAmnali'nsdivbiodduyalweidgohste HHI 6329-133 o(2n.t0heal/dkagy)ofwastrecaatlmceunlta.tedThaendcomnetarsoulred based ofstearpiplreoxiwmaatteerlyfo0r.0i5njmecLt/iconn.) was applied evenly to the test site amsaterrsoonls rS1r2eo:w8rnsmig/2,kuga,t3e.rraensfdpoerc4.tiinvjeeFlcoytr)i,otnhetthoeGratoeuscptosncm2ae,tnet3rr,iaatalinodn(4To-f6a0n59i49m,)als9w5a0s(,0m.i1"ax2n8ed,d9.w19i.2c20k8,ng oytneheersepitersceststiopvreeecldtyi,vaet atnredosotmapmptaletimeeprdeiraaalttumraeixdtouunsrteielvwoalasudmmceianliocsftuel0ra.et0de1.d mlfA/oknrg.eiancdhiTvhieadnuimamilaxleduboracncse/ymolf, of body exposure weight op was 4 cn` 0.006 l/c'. the and day the of treatment. For approximate rate of all three groups. application was the sea 1r0ocump x5. 10T-hcea tseescttiomnateorfiamlate(rTi-a6l051t)hatwaswasapmpoliisetdenteodewaicthhadniisntaill'lseyskainnteass. a SEpacchureadrewaitohf Elastoplast tpaaappppeelrictoattappieroonvairwdaoesundacnovaloelcrceldeudsgwieivstehadnarde1so0sv-iecnrmgw.rxap1pC0oe-ldclnawrigstahuwzeeSraerbaanunsdeHadrgeapvo and restrain the animals during the 24-hour exposure period. wyAaepssptrroraxeimionmviaentdgelwyciotl2hl4arhtsaopuraswndataebfratnedraandgteesdsitswpeoorrseacborlneetmroovpleadpmearatnedtroiwaaenllys.raepspildiucaaltiotne,st tmhaeterial Reason for Route of Administration The dermal route is a potential route of exposure in humans. Observations of Animals cooblnsihdenouriuccvrtaaseltdiaofondtbsesreravntdaetsittowniocsre wcoea nrtedraoyclomnomdaruttceatrleiidatlypreachddemocisknesisatn(rdmaotraintoinna:gpparnAoddxdiimatfaittoeenrlanyloo1n,C)o2nw.i5ec,aeland aily thereafter for 28 days. mBaotdeyriwaelighatdsninwiesrteradteitoenrmi(nDeady f1)o,r arnadndoatmiziant-i1oinfeputrepromsiensatioonnDa(ybay-9,39)b.efore test [mTaeahdaemrdiiinieninsdsi)tt.riaaaflticoodSneurbmsaaecqlcuoernidtrirnrigetaadttoiinogntsheroefaDrddaieinzrgen'aylatseicmrharndiietqautbeieofn(orreweceortreedsetmdadoaers actpohpnertrooDxlaiynamta1etleyria3l0 GCrRoouuphls4 uwaasstertihneabdavDneadyratgee8nterlreyyntovhnaeolmtar(esDccaoyorrde2e)df.oarndonaen Dfaeymsale4 aannidna8l. (TNoh.e Foonzldysse)xcairtion 00286 Page 12 of 46 Sample Collections HHI 6329-133 bF1Bl5ll,oagooodadnwisanaasmlpscloensbdeuf(coatrpeepdreoxaxppiepnrraiotmxeeilnmytaat4lelnyli)n2i4wt-eihraoteuirocsnolpl(oeDsactytdeod1s)e.fro(SnDuaybasem2)aq.rugeinantnadlcoonelaiDraeyvcsehien ooff Tj2ha0emspmelaensd2w2eD2r.oeodcIennwtaarsdidfoiutbgitoeandi,neadnadtfrstoehmeparttahietmeedpoosiftnetnoreicosrreorpusvnyenaaonndcaDvcaaeyllo2fu3.laaerapcphfrroaaxcnitimimaoetnss"e.ly4",118, -20C s1a0mpClesuntwielreshtihpepnedstotoretdheinSpaonfsroere.zer set to maintain 3 tansersvae of Pathology faAnontwddlgineceeracomrdioinpuasntiieopdnentioonfbartrahbneidtoeamxlp,oerrdibemlree.ndtavliThaephtsahiseteepso(sDtoafeyrti2eo9s)r,t veaannnadimacclaovsnat,rwoelreexmsaaatnnogeeusiitnnh:getteisz:ed All any aabnnitonraimloasnliwwteeirreees wswauesbrhjeeecdtreewdciotrhtdoeldau.nkeanTbahbrernewvthiaoaplteewdaltiegvrreorsb,sefobnrielecer,atphasenyneaecpxrparomopixsniyamtaiptorenocecdousre. PKFhli1iadsncnseeudyexs slafomrcponlnessethcweteirofenirwsoetfigmthahelede dea(rnvndoallufneemaapolpnelliycian tdieeotancehrmsiginrteoedupffrwooermrebailllceo)lalneiacmntadeldsi,.nmesJdnud|eatbeoltyh "20C 1o0nC.d"ry Aifcte,ertnheencropplsayc,edthien aanifmraeleszerwerseetditsocamradiendt.ain a tenperator cf Shipment of Blood, Bile, and Tissues 2Ajf-rfr3ota6gce-etr0ni9oe,nx(spo)ne,ridmrleyinvteiarclse,) tetboirlmeti,hneatdieSorpnno,anlsotrhaep(pJlbailmcoeaostdiDos.namJspoilhtenesss,o(ns,aenrdSuHnkiEa.dnEnd.eycs4elpwl.eucrl.aa,rse1ndtg. wdanieasilpgyohsstiisstioor9n3v5oolfBuumstehhse.AvseanmTuphele,e'sS.pSto.nHsHoPIrauldi,osesMrNe,snopto5n5s1a0ic6bc)le,pet faaolnroyngthreewistrpheotnestnihtbeiiiolrinctoyranrdfeosrpotnhdeing presentedofinthtehistirsespuoerts.amples collected in this study nor are these rescies Statistical Analyses No statistical analyses were required by the protocol. Location of RawData,Records,andFinalReport rTheetairnaewddaitna,therecaorrcdhsi,vesanodfanHHIoriigninaaclcorsdiagnnceed wciotphy HoWfI tSheOPfinal report will be 600287 Page 13 of 46 Body Weiahts RESULTS Individual and mean body weights are weight gains from Day 1 to Day 29. in Table 1. HWI 6329-133 A11 animals exhibitedbody Clinical Observations Individual throughout clinical signs the study with are the in Table 2. exception of All one animals appeared normal female animal (No. F52900) treated with T-6054 the last 22 days of at 1.28 study. mg/kg that exhibited This animal also had weakened hind 1imbsduring small feces on Day findings are considered to be collection procedures and are due not to an injury incurred during the 8. These sample considered to be test material-related. Dermal Irritation tIensdtivimdautaelriadlermTa-l605i1rrpirtoadtuicoen scores are in Table 3. The control material and observed in the animals treatdednowidtehrmaTl-60i5r4ritatatiaond.ose Nolevdeelrmaolf irritation 0.128or was 1.28 mg/kg. T-6054 produced slight to and 4 only in five animals at the 12.8 moderate erythema mg/kg dose Tevel. reactions at Days 2 Pathology Individual animal pathology comments are lesions observed in any of the animals. presented in Table 4. Page 15 contains a Pathology report by the study pathologist. There were no DISCUSSION The and acute systemic absorption/toxicity and relative T-6051 were evaluated in male and female albino skin irritancy of T-6054 rabbits when administered as a single dermal result in any test maaptpelriicaatli-orne.lateAdppleifcfaetcitosn onof itnh-eliftehescelinmiactaelriaflisndidnigds,nobtody weight gain or test material macroscopic findings T-6051 did not produce at necropsy. any dermal The control irritation. material No dermal and ti1h.re2r8it1am2gt./8ikogmn.g/wkagsT-6do0ob5ss4eervpleervdoedluw.cietdh vTe-r6y054sliagphptlieddermaatl a idrorsietatlieovnel inoffi0v.e128anoirmals at 00288 . Page 14 of 46 SIGNATURE HHI 6329-133 Steven M. Glaza Study Director Acute Toxicology Date o=\lo-Ax REFERENCE 1. Dorfaitzhee, SJa.fetH.y, of"AcCuhteemiDcearlmsalinToFoxoidcsi,tyDr(uSgisnglaendECxopsomseurtei)c,s" - IDn:ermaAlppraisal Toxicity, (1959). Association of Food and Drug Officials of the U.S., pp. 54-56 00289 Page 15 of 45 HHI 6329-133 PATHOLOGY REPORT TlTnhetevererelisaWlee,urtehdoassnieixzelrdeavbebalin,tdsndae(yctrhoorfpesedieemadatlhea,st aninal are athned and gttrehorrsmesienaoftbeismoeanrlveaso)tfiosenhasechrsetfucrdooyrn.defdiTvhedeotseest in the Individual Pathology Comments that follow gsrrepomrotr.each bAbaoltirlleen,aencciroamontapelsacsypt,,proatnxhdienrbaeottewher1ke-icdnnnoexyvsi1s-ifcbrnloensectTtheeisoinfoinrsosftitnmhaealnedyeraomnfadltfheesmppsa]lneiimraaalsi.sonnchsTgihrteeToifvreorm, dbiilsec)a,rdefdr.ozeend,. andThesenttisstuoethseamSpploenssowre.re Awfetiegrhednec(rvooplsuym.e anly determinupPor the'acsroywere (thaoslogEi.st.Palmer Pho; 0(361302995-133.51h) ate6-16-95 G00290 Animal Number [foeeTs FS2898 Mean [[3s2e89r3 FS2897 Mean [Foeaeosls F52892 Mean [52288880 FS2899 Mean Page 16 of 46 HWI 6329-133 Individual and TMaebalne B1ody Weights (g) RandoHma-le ization __Day RandonF-emale Animal ization Day Day-9 TI zm Number Day -9 TT 29. Group 1 (Control) - Sterile Water for Injection (0 ma/kg 2L229024 2,169 22,34803 227,4855 FE5S29i7 2,366 2,745 F52883 220,303018 2,274 a2l,e3v6e8 22,,987683 5l408 2,985 2,222 2,330 2,781 2,20 2,447 2,082 Group 2 - T-6054 (0.128 ma/kg L2,221024 2,338 22226308 2g16e03 FF52208716 2423 2913 Fs282 22,2m70 212 523,6380 22,,884329 3a 2,814 2,271 2,306 2,712 2,226 2,300 2,832 Group 3 - T-6054 (1.28 ma/k 22,20187 2,261 22,.338639 227,5976 FF9s20ee04 2,333 20614 F598 22,.325010 2.166 s2l,a560o8 2,2,598288 5.089 2,822 2,195 2,362 2,779 2,242 2,419 2,779 roup 4 - T-6054 (12.8 m L2,026816 2,353 22,241197 22,572953 FFa5m9e4d 2,393 2,741 F585 2z,ii0g000 2,205 3d5e7 22,,885337 lees 2,791 2,267 2,383 2,686 2,224 2,43 2,827 00291 Page 17 of 46 : Animal Mumber F52879 F52963 F52874 Mean HWI Table Individual and 1 (Continued) Mean Body Weights (g) Male Randon- RandomF-emale ization Dav9 Day TTT 29 Animal Number ization bay-o Day TTT Group 5 - 7-6051 (10-cm x 10-cm Section 2,278 2,173 2,048 2,398 2,258 2,923 2,622 F52877 F5888 2,021 2,191 2,265 2,495 F52966 2,157 2,273 2,264 2,322 2,166 2,307 2,680 2,159 2,251 6329-133 am 2,782 2,752 2,792 2,775 @ Animal No. for use in Fth5e289s0tudwyasbuotriwgaisnalrleyplasceeldecwtietdh bNyo.theF52r8a7n7domdiuzeattioonpooprroghreaalmth. 00292 Page 18 of 46 HHI 6329-133 IndividualTaCbllieni2cal Signs Animal Sex Number _O_bservation 1-4 Hours v1) Day 7-7 8 Group 1 (Control) - Sterile Water for Injection 5-37 (0 ng/kg Male FF5522887835 AAppppeeaarreedd nnoorrmmaa]] F52808 Appeared normal vv or y y 7 ov Female F5522897687 AAppppeeaarreedd nnoorrmmaa]l F52883 Appeared normal vv v o7ry 7 ory Group 2 - T-6054 (0.128 ma/kq Male F52887 F52803 AAppppeeaarreedd nnoorrmmaa]l v 7 F52897 Appeared normal v Female FF5522897061 AAppppeeaarreedd nnoorrmmaal] F52882 Appeared normal vv 7 o vor u y oy o vor r y ory Group 3 - T-6054 Male FF5522899615 AAppppeeaarreedd nnoorrmmaall F52892 Appeared normal Female F52884 Appeared normal F52900 WAepapkeeanreedd nhoirnmdallimbs small feces F52968 Appeared normal (1.28 ma/kq, v vv v v v voor u y I ov I SvT =v vou /- CCoonnddiittiioonn enoxtisteevdi.dent. 00293 Page 19 of 46 IndiTvaibdlueal2 C(lCionnitcianluedS)igns HAI 6329-133 Sex NAunimmbaelr Observation 1-M4avHo)urs 7-7 8Day. 5% Group 4 - T-6054 (12.8 ma/kg Male FF5522888806 AAppppeeaarreedd nnoorrmmaall F52899 Appeared normal v v ov ov 7 voor Female F5F522889849 AAppppeeaarreedd nnoorrmmaall F52895 Appeared normal v 7 vor ory 7 [A Group 5 - T-6051 (10-cm x 10-cm Section' Male FF5522986739 AAppppeeaarreedd nnoorrmmaal] F52874 Appeared normal vv uv y y 7 ovr Female F5F522888787 AAppppeeaarreedd nnoorrmmaall F52966 Appeared normal v7 o vr oy y 7 oy 7 Condition existed. G00294 Page 20 of 46 HUI 6329-133 Individual DermTaalbleIr3ritation Scores Group 1 (Control) - Sterile Water for Injection (0 mg/kg) Males Females DermalReaction ---- StudyDay 124 --StudyDay 2 4 2 43 Anima) Mo. F52885 Aninal No. F52967 EErdyetnhaena Atonia 00 00 00 00 0000 00 00 00 oooo 000 CDoersiqaucaemoautsinoenss Fissuring 00 00 00 00 0000 0 0 0 o0o 00 00 00 0 oo EErdyetnhaema DeAstqounainaation CFoirsisaucreionugsness Animal No. F52873 00 0 0 0 0 0 0 00 0 0 0 0 0 0 00 00 00 00 Animal No. F52878 0 0 0 0 0 0 0 0 9 0 0 0 0 0 oo o0o 00 00 09 EErdyetnhaema ADetsoqnuiaamation CFoirsisaucreionugsness Animal No. F52898 00 00 00 00 00 00 00 0oo 00 00 00 00 Anima) No. F52883 00 00 00 00 00 00 00 00 00 00 00 00 G00295 Page 21 of 46 IndividuaTlabDleerm3al(CoInrtriintuaetdi)on Scores HHI 6329-133 Group 2 - T-6054 (0.128 mg/kg) Males DermalReaction-- T= S2 tudyD4ay 8 EEdryetmhaema ADetsoqnuiaanation FCoirsisaucreionugsness Anina) No. F52887 00 00 00 00 00 00 00 00 0 oo 00 00 00 Females ---- 1 Study Day 2 3 Animal No. F5290: 00 00 00 00 00 00 00 00 00 00 00 o0o EEdreytmhaena ADetsoqnuiaamat on CFoirsisaucreionugsness Aninal No. F52893 00 00 00 00 00 00 00 o0o 0oo 00 00 o0o Aninal No. F52876 00 00 00 o0o 00 00 00 o0o 0 00 00 0oo EErdyetnhaema DAetsoqnuiaamat ion FiCsorsiuarcienogusness Aninal No. F52897 0 0 0 0 0 0 0 oo 00 00 00 00 0 00 00 0oo Aninal No. F52882 00 00 00 o0o 00 00 00 00 0 00 00 o0o 00296 Page 22 of 46 HWI 6329-133 Dermal Reaction Erythema Edema Atonia Desquamation Coriaceousness Fissuring Table 3 (Continued) Individual Dermal Irritation Scores Group 3 - T-6054 (1.28 mg/kg) -- -- SMavleDsav T 2 4 = TSThFeu nalpead sy y IT --AnNio.n52a96l5 _nina) Mo. Fssss [J 0 0 0 0 0 0 [J 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F5289] 0 0 0 0 0 [J 0 0 0 0 0 0 0 0 0 0 I 0 0 0 0 0 0 0 Animal No. F52900 [J 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 [] 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52892 0 0 0 0 0 0 0 0 [J 0 0 0 0 0 0 0 0 0 0 0 0 0 J 0 Animal No. F52968 0 0 0 0 0 [ 0 0 0 0 0 0 [1 0 0 0 0 0 0 0 0 0 0 0 600297 Page 23 of 46 Table 3 (Continued) Individual Dermal Irritation Scores Group 4 - T-6054 (12.8 mg/kg) HWI 6329-133 Derma) Reaction Erythema Edema Atonia Desquamation Coriaceousness Fissuring Males I 2Study 4Day Animal No. F52886 0 0 0 0 0 0 0 0 0 0 0 0 0 0 I 0 [1 0 0 0 0 0 0 0 Females I og Study Day Animal No. F52889 0 1 1 = 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 I 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52880 0 1 1 I [J 0 0 0 0 [J 0 0 I 0 0 0 0 [J 0 0 0 0 0 0 Animal No. 0 1 0 0 0 0 0 0 0 0 0 0 F52894 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52899 [] 1 0 [3 0 0 0 [] 0 0 0 0 0 0 [] 0 0 0 [3 0 0 0 0 - Value not recorded. Animal No. F52895 0 2 1 0 I 0 0 0 0 [J 0 0 I 0 0 0 0 0 0 0 0 0 J 0 . 00298 Page 24 of 46 Table 3 (Continued) Individual Dermal IrritationScores HWI 6329-133 -- tales ene Group 5 - T-6051 (10-cm x 10-cm Section) DermalReaction -- I~ StudyDay 2a 8 -- Study Day _ 1 2 am Animal No. F52879 Animal No. F52877 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 0 0 0 0 0 0 I] 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 I] 0 0 0 0 0 0 0 [ 0 0 0 0 0 0 0 I 0 0 0 [J [J Erythema Edema Atonia Desquamation Coriaceousness Fissuring Aninal No. F52963 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 AniNom .Fsa2as)s 0 0 0 0 0 0 I 0 0 0 0 0 0 0 0 0 0 0 0 I 0 0 0 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52874 0 0 0 0 0 0 0 0 0 0 0 0 [] 0 0 0 0 0 0 I 0 0 0 I Animal No. F52966 0 0 0 0 I 0 0 [J 0 0 0 [1 0 0 0 0 0 0 0 [ 0 0 0 [1 600299 ' Alnuimmbaelr F52885 F52873 F52898 F52967 Fs52878 F52883 F52887 F52893 F52897 F52901 F52876 F52882 Sex Group. M M MN F F F M M M F F F Page 25 of 46 Table 4 Individual Pathology Comments HWI 6329-133 Died TesStacDrayificed NecropsyObservation Control) - - Sterile Water for In, lection (0 mg/kg. 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. Group 2 - T-605 (0.128 po/kq - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. F52965 M - F52891 M - F52892 M - F52884 F - F52900 F - F52968 F - - Not applicable. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 600300 Page 26 of 46 MAunmimbaelr F52886 F52880 F52899 F52889 F52894 F52895 F52879 F52963 F52874 F52877 F52888 F52966 Table 4(Continued) Individual Pathology Comments sex Died esStacDrayificed MecropsyObservation Group 4 - T-6054 2.8 mg/kg] M - M - M - F - F - F - 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. Group 5 - T-605] 0-cm x 10-cm Sect: jon; M - 29 No visible lesions. M - 29 No visible lesions. M - 29 No visible lesions. F - 29 No visible lesions. F - 29 No visible lesions. F - 29 No visible lesions. - Not applicable. HWI 6329-133 00301 Page 27 of 46 Table 5 Individual Animal Tissue Weights and Bile Volumes HWI 6329-133 Weight Sex AMnuimmbaelr Liver Kidneys Group ontrol) - Sterile Water Male F52885 70.634 - F52873 75.513 16.417 F52898 74.433 - Female F52967 62.517 - F52878 63.222 16.808 F52883 75.418 - (q; cDeartmiaolnApSpiltie- for injection 0.960 0.866 0.645 0.777 0.890 0.618 VoBluimlee (ml) ma/kq. 1.0 1.6 0.9 2.1 2.0 2.5 Group 2 - 71-6054 (0.128 ma/kq' Male Female F52887 F52893 F52897 F52901 F52876 F52882 71.149 64.537 73.622 71.908 66.359 77.959 13.509 14.610 - 0.784 0.836 0.604 0.640 1.179 1.337 Ll 0.6 1.5 L7 1.2 1.6 Male F52965 69.880 18.191 0.452 1.8 F52891 65.695 - 0.525 1.1 F52892 69.216 - 0.949 1.1 Female F52884 65.907 - 0.590 1.2 F52900 61.937 - 0.710 1.0 F52968 67.347 12.513 0.866 L3 = Not applicable. 00302 Page 28 of 46 Sex Male Female Male Female Individual AnimaTlabTliess5ue(CWoenitgihntused)and Bile Volumes HHI 6329-133 Animal ---- Weight (@)Dermal AppTi- Number Liver Kidneys cation Site Group 4 - T-6054 (12.8 mg/kg Bile Volume (mi F52886 77.219 - 0.857 0.6 F52880 50.985 12.254 0.634 0.8 F52899 89.883 - 1.301 0.4 F52889 69.288 - 0.953 0.5 F52894 75.086 - 1.000 1.4 F52895 59.174 15.603 0.635 0.9 Group 5 - 7-605] (10-cn x 10-cn Section F52879 89.874 - 0.884 10 F52963 71.814 - 1.535 0.5 F52874 71.657 15.784 0.990 0.5 Fs2877 71.284 - 1.154 0.5 F52888 65.633 - 1.231 0.5 F52966 68.719 15.855 0.898 1.5 - Not applicable. 00303 Page 29 of 46 APPENDIX A Protocol Deviation Protocol TP3016.AB Protocol Amendment No. 1 Protocol Amendment No. 2 HWI 6329-133 600304 ) Page 30 of 46 Protocol Deviation HUI 6329-133 Protocol CP.ageObs7e,rv7.atiEoxnpeorfimeAnnitmaallsD,esi(g2n), RSeeacdoinndgSeonfteDnecrem.al AIdrdriittiaotniaoln, bdeermmaaldeiarprpirtoaxtiimoanterleyadi3n0gs will minutes (Day 2) aanfdteorn bSatnuddaygeDaryesmov4al and 8. Actual Procedure. oiTnhneeadGvDreaoyrutpe8n4telryfyetmnhaoetlnearesccoorrdeedwasfor (No. F52889). oThfisthedevsituadtyi.on is not considered to have had an adverse effect on theoutcome 00305 Page 31 of 46 ) JOSIE AR FRAT > CORNING Canny Sponsor: St. Paul,MnHinnesota PROTOCOL TP3016.48 Study Tite: Single-DosTe-6D0e5r4maalndAbTs-o6r0p5t1ioinn/TRoaxbibciittsy Study of Date: ) Decenber 13, 1994 Perforning Laboratory: Ha33z0l1etoKninsWmiasnconBsoiunl,evaIrndc. Madison, Wisconsin 53704 aboratory Project Identification: HHT 6329-133 ) HESS Ma ORUVERY 00306 nus inman mov wean on a LE Page 32 of 46 ) TP3016.48 Page 2 STUDY IDENTIFICATION Single-DosTe-6D0e5r4maalndAbTs-o6r0p5t1ioinn/TRoaxbibciittsy Study of HI No. 6329-133 Test Materials 1. T-5054 2 T-6051 Sponsor 3MDeTpoaxritcmoelnotg.y Service Medical 3P.M0.CenBtoexr,332B2l0dg. 20-26-02 St. Paul, WN 55133-3220 Sponsor's Representative 3JHohnToxL.icoBluotgeynhoSfefr,vicPeh)Medical ) 3HDeCpeanrttemre,nt.Bldg. P.0. Box 33220 20-26-02 S(t6.12)Pau7l3,3-1W9N6255133-3220 Study Director HSatzelveetnonM.WiGslcaoznasin, Inc. MPa.d0.isoBno,x W7I54553707-7545 (608) 241-7292 Study Location BHuaizlldeitnogn NWoi.sc3onsin, Inc. M3a8d0i2soPna,ckWeIrs 5A3v7e0n4ue PrEoxppoesreidmeSnttuadly TStiamrettabDlatee Experimental Termination Date JDaencueamrbyer252,8, 19199594 Draft Report Date Harch 8, 1595 ) 00307 Page 33 of 46 i PTaPg3e0136.28 1. sStiundglye-Dose Dermal Absorption/Toxicity Study in Rabbits 2. Tpourpdossseess Irritancy otfhetessytstmeamtiecriaablssorwphteinonapapndlietdoxitocitthye asnkdinreolfatriveebisk.in 3. ReLUahgbruoelrraastttooirrdyyyCwPoirmlaplcltiibceaenccRoenedguucltaedtiionnsa/cSctoradnadnacredwsi,tGhuitnhdeeesl.following Good [[X1] 2C1onCdFuRct58as(FaDAN)onregulated Study [['1] 4400 CCFFRR 719620 ((EEPPAA--TFSICFAR)A) [[11 [1 N5o(98t1io)fh3i0sc"aant(iFoNinon.aNlo)3.85(3001E3c"(DJ)(aJpaapnaenseeseMAFHFO) ) wddfoiierk]rl.efcahtrroeorc,AecdtuthreeResgsutilunadtytihodinosse.'spronotItoncoutlnhneeacroeepsisniainroincloymopfdluitpahlenicceSaptewe.intshenrythapnedeAenisiusncsalyl ) 4. QtthhueealihQutuoaytlgiAetseyslu,ArsasnsuctreuadyncecoUnndiutct,inanadccotrhdeanfcienalwitrhepoHratzlewitloln be audited by (HNT) Standard Operating Procedures (SOPs) and pol seins.Hisamtae 5. Test Materials A. I1.dentTi-f6i0c5a4tion 2. 1.6051 B. P1h.ysi(cTaol beDedsoccruimpetnitoned 2. (To be documented iinn tthhee rraaww ddaattaa)) C. daPenutereirtSmypionnaasntodiroSnatssasbui(nlieinstcylrudeisnpgonsuinbdielrittyestforconpduirtiitoynsa)n.d stability 0. RStooormagteemperature ) 00308 Page 34 of 46 ) PTaPg3e0146.48 E. RReesseerrvvee Ssaammppllee(ss) materials will be tofakeenachforbattchihs/losttudoyf. test and control UJNiIe for, iLnesta afnrdeezcearntrsoelt 10 years per WWI mStOaoPt.emraiianTltheairnSepsoeanrvsteoermpsweairmlapltluerbseewioclfolnt-ab5ce0tesdteorregdoat r$e0guyleaatrosryforGododispLoasbiotriaotonryinPraaccctoircdeasn.ce with the sppropesatsnotor F. Retention Aconmypluentuisoend otfestthematienr-ilailfse pwihlalsebeofrettheurnsetdudyt.o the Sponsor after 6. SaAsferteyqPuirreecdaubtyioHNnIs SOPs and policies 6. ControlMaterial A. DIidsetnitlilfeidcatwaitoenr , 8. PhCylesairc,alcolDoerslcersisptTiioqnuid C. TPuhreitpyuriatnyd considered t"Saotndabbesitlaaidbteiyqluiattyeofforthitshemapnuurpfoascetsureodf mtahtiesrisapludyi.s 0. RStooornagteempCeornadtiutrieons E. RSeeseerSveecStiaomnpl5e.s. Reserve Samples : F. Retention Aanndy wrielmlainniontgbecondtirsoclardmeadteraifatlermaiyssubaencueseodfftohreotFihnearl treesptoirnsg. 6. SaAsferteyqupirreecdaubtyioWnWsI SOPs and policies 7. Experimental Design A. Animals (1) SRapbebciites ) (2) Hsetar:ai(nN/ISM)ouSPrFc/eHRP, Inc. . 00309 Page 35 of 46 ) (3) AgIenaittiation Adult (9) YeataInih tiattion 2.0t0 3.0 kg. PTaPg3e0156.A8 (5) Number and Sex 15 males and 15 females 0) Husbandry (6) Identification Individual numbered eartag (2) Housing Individually, (heavy gauge) in screen-bottom stainless steel cages (0) Food A measured (PMI Feeds, amount Inc.). of Laboratory The food is Rabbit Diet HF #5326 routinely analyzed by . etnhevimroannmuefnatcatlurecrontfaomrinnaunttrsi.tional components and ! (e) ater Ad Tibitum from an water are analyzed automatic system. Samples by HWI for total dissolved of the solids, ofhraogrradnnsoeesplshe,ocstpeahdnadteelssep,meecnaintfdsi',ecdhhlemoairvciyrnoabmteietoadllosgh,iycdarlocacrobnotnesn.t and (@) Contaminants tThhaetrewoaurlednoinktneorwfnerecownittahmintahnitssstiundyt.he food or water Tight/12-hour dark cycle. (e) Environment sEentvirtoonmmaeinntatlaincoantrtoelmsperfaotrurteheofani1m9alC room will be to 23C, a relative humidity of 50% 20%, and a 12-hour (8) Selection of Test Animals (f) Acclimation At Teast 7 days Based on adequate health number and body of extra weight animals according will be to HNI SOPs. purchased so An that no animal in animals will obviously be placed poor into health is placed on test. study groups using a The ) dsatyrsatioffiesdtudbyodyiniwteiiagthitonr.andomization Program within nine 00310 Page 36 of 45 ) TP3016.28 (9) fJhuesttoirfiiccaaltliyo,n the animal "ftohre NSepweciZeesalaSnedleWchtiitoen albino Page rabbit has 6 been background oifnfcohromiacteionbeocnaustheisofstpheecileasrge aman on B. DAodsmeinistration (1) Test Groups Grow Test Material Dos(0ea/Lkeav)el NMalues mofbAFneeimmaajrlsss 21 (Control) DisTt-il6l0e5d4water 3 T-6054 0o.v128 33 33 45 TT--66005514 121..828 33 - 3 33 3 **+ JToo bbee aaddnmiinniisstteerreedd material (fabric) aats 2a d1o0s.e0-cvnoluxme10.o0f-02n.0pmriockegof test | (2) gP0ia7fcrhntehreceaebsdpbsaiaytrayoBefrf(EotorxaepootbtsteuasiritenAomaurtnneebrlaieamlishaepdplisckaitn)i,on,thetheFlboaccek aand, 2S{o0iu%nptpaoecfrt.,thsekiTnht)eowtialsllhabvboeeddyclasireuparpfeadwcileflraereceoano.sftihtTauhietretwiratephsptrmoaenxnitnealtceaisittbyosrc otShfFrTotiuhtgeahtoiauontni,malosr.widlelfTehcebtesaniitnmhsaapltescwtwoeiudllldfobprereiccnlltiuepdrpefeedtrhieansgusnteee.esdonfadmes:hy the study. (3) DPJ1eo1ssheeacAntdiimmvaielnsisttewsirtlaltoirorneccoenitvreola wil msaitnegrlieala.dmiTnhiestdraaytioonf tofrestrhecent aanniimmaalblsesdibneosdiGyrgonwuapetsiegdh1t,asj2,uDsat3y,'b1e.afnodrTe4heawidrlaelisnpibesecttribavatesieoddnosoensthgear she Ty9nhndetiol,Gutrthoeuedp.ar1,eaTh2eo,f3G,reoxuappnods5ur4meamtaietnreiraailatlhswiinlwlia1nldbe ubaenpipfalopiprelmdieaasnvders"pread W1{iG0tr,ho0u-pcdsnist1x-i5l1)l0e.dw0i-lwclantebprei.'eccoevTehorefedatrhweeiatthoefsta amp1ap0tl-eicrnciaatx)io1nm0-ocsnsoegsoanuadze crPDorealonrlvduaiarrdgaeeepdpasenedcoucwrcietldhuswiiSvtaehrandpraeWprseasrpintga.apned TaEhrleoausrntadobpp3li1at1sstewdigttea}spepaentyo J) during the 24-hour application pening. 00311 [-- ) ) Page 37 of 45 TP3016..A8 (4) Reason for Route of Administration Page 7 h[uhmeandse.rnal route is a potential route of exposure fn (5) RAepmporvoaxlimaotfelTyest23Mahotuerrsiaafter application the bandages and ctoelsltarosr wciolnltroble mraetmeorvieadl atnheaprpesriodpurailatetesstolvmeantte,riailf nwielclessbaeryr.emoved using warenanodr C. Observation of Animals (1) aCFldoimrniinccilaislntircaOatblisoensrivgaanntsdiofbnoesrforcelintiecsatl orsigcnosntraonld mmaotretrailailty aSaldpompnirsno,ixsitamrnadatteiltowynic1e,(Dda2ay.5i,l1y) aan(nda.dm4.dahiaolnuydrspt.hamef.rt)eearfftoterersmtofrmotaratlecrilitiayn)iactal daitrelcetaesdt b2y8 tdahyes.stuOdybsdeirrveacttioorn.s may be extended whey for (2) dRBeeerafmdoairlnegitoerfsrtitDaoetrrimoacnlontrIreroarldiitnmagattieworinlilalbeadmmaidneistarndatiroencortdheed initial DaIfartyreirt1 atbriaeonanddaignregeadr(ieAnmtogtvsaaclwhimle(lnDtaybe12)).madaneAdddaioptnpirooSnxtaiuldmyatdeDealrmyyas]304 as the mainndut5:es fIonrdiveiadcuhalanidmearlm.al irritation records will be maintaiueq (3) aBFpoopdrlyircaHanetdiiogomhnitzsa(tDiaoyn,1),beofnorDeaytes2t9, oarndconattroulnscmahteedruilaeld death (when `survival exceeds 1 day) (4) Sample Collections (2) EBerfeoqrueenciynitiation (Day 1), post-dose (Day 2), Days 4, 8a,ppr15o,xim2a2,telanyd 24at hours experimental termination (Day 29] (b) ANunmber of Animals (c) MBflerotonohdodthseaomfmpalCreogslilne(acaltpipoernaorximvaetienlyof4emiit)herwilelarboencDoalylsectI.ed {2,act4,ual8, vo1l5,umeandto 22b.e be obtained from the dpAoopcsputmreeronixtoiermdavteeinlnaytch2ae0vamrLaowfodfastabacl)hoowdill animal sacrificed in a moribund condition or 00312 ) . ) J) Page 38 of 46 PTaPg3e0186.28 csseaanmctprrliieffsiucgweeiddl,latabnetdhestthoteriendseepoaatfranrtoeeocmrospetsreyummpe(arDnaadtyucr2ee9l).]adnydaTrthheen ffSr2ra0acctCtiioon1ns0sCws.itlolrTebhdee isnseenpatarfafrtreeoedzzeenrseorsnuemtdratynodmiacceienlttloauilsatrhe Sponsor after experimental termination: Samples will be shipped to: JMamEesE0. 8 JoPhCn.son B9l3d5g.Bus2h-3A-v0e9nue St. Paul, MN. 55106 JtthaeemleespshamoD.pnleeJsoa.thns(o6n12)or77a8l-t5e2r9n4ateprwiiolrl tboetnheotishfiipemdenbt.y of D. Pathology (1) UnAmsoncryihbeaundniudmlaelcdondSdyaiictnrigiofndiucwreiislnlag nbtdehDeesusabttjuhdesycteodr tsoacarnifiacbebdreviinataed garnreeocssortsdheendte.iczreodApnswiyimtaehlxsamsoiindniaautniomnopreinatbnoudbnadarlblciotnaadblintoir(omvniaalwiitilinlejsebcatwiilo]n be in Det.xhseaPnamgtauhriognliaontgaeyld,.ea(3r)TivsSesianum)ep,sl,e bClaoseldldeevcistaciroitnbh,eedwvieilnnla sbceeacvteci.oolnlsencdted. After necropsy, the animals will be discarded. (2) AsStucrhvetidevurilmneigdnataSinaoicnnrailofsficwetihlel exbeperainmeesntthaeltizpehdasweit(hDaysod29i)u,m baplbeebndrteovbviairaabtiettdhaelgrvoe(snvsaiancaeivcnarj,oepcsteyixosnaenxgaiumniinntaahtteieodmn,a.rgainndTahlesuaebnajiermcatvleesdin)w1,i6l)an bbee rneeccorrodpesdi.ed in randon order and all abnormal ities wil (3) wSTaahsemhpelsdeitweCisotlhlofecltutikesoetnwaranndtacpontwraotlermaptreiroiraltoatphpelinceactrioonpswyill be ap1nr-iocmcnaeldssue,rcet.iaonnd Tohbefothwthhokelieddenreimyvaselrf,raopmpblitliehc,eatifaoinnrsatpsipmtraeolxeifmroaanmtde alf1leomcanlex pnleaccoepdsieidn ainferaecehzergrosuetp 10C. After necropsy, wtoillmaibnetacionlleactteedmpearndatuirmemeodfiatge0liyc the animals will be discarded. 00313 Page 39 of 46 : P3016.48 Page 3 TkShipedonnestyoisrs)suawefistlelr(lebixevperes,reinntbeinlftera,olzdeentremroamnlindraaytp.ipolinic.ecattiToohnethessaimtep,les w7ri.elCtl.e(n4tb)ie.o(nsch)ai.npdpeddTihsetpooSsptiohtenisoopnrerosifsonrthelesipsostanemsdpilbielnse.Sefcortiotshe E. Statistical Analyses Wo statistical analyses are required. 8. RAepfoinratl report including those items listed below will be submitted. DDeessccrriippttiioonn ooff tthhee tteesstt asnydstecmontrol materials TDPaarbtoueclseadtuoirfoensexopefrimmoernttaallityindiattiaatibyonseaxndantderdmoisneatiloevnel TGDraeobssucslraitppiatotinhoonloofogfymeaanfnyindbtioondxgyiscw/egeirfgofhsestcstsp/abdtyehrosmleaoxlgyanirdrerpdiootrsatetiloenvel ) 9. LoOfIracsiceagiptiltiniaoatnnlacoteedfatoRaafua,dwitDhtoeirantgcFaion,ptaiRhleeescrsoettrpuhdodersyrte,.oadfun,rdiwFnigilnliatlsbeRperapovogarritelsasbleandatbeHfIoreto 2r1e1taionreidginianl thpeapearrchdiatvae,s oifncHlNuIdinagWchceotrnhodsitenhgeittfoeinnaH]H1IisrtSeoeppdo.rtbelioswcwormtp]letoeed, DPorsoetocporlepaarnadtipornotorceoclordasmendments InB-loidfyeweriegchotrsds ODbosseervaadtmiionnisstration ASnaamtpolmeicCaolllepcattihoonlogryecorredcsords SSthuidpypincgorrreecsoprodnsd.ence Final report (original signed copy) Tnhoet bfeollaorwcihnigvedsuwpiptohrtitnhge srteucdoyrdsdatwai.ll be retained at HHI but will AWnaitmearl anraelcyesiipst/arceccolridmsation records ARInenifsmrtairlugmeerrnaottoomcratlaeinmbdprearftariteouenrzeeranadntdemmaphieunrmtaietdnuiartneycerreercceoocrroddrssds ) 00314 Page 40 of 46 3 TP3016.48 Page 10 PROTOCOL APPROVAL Phe 4.Built John L."Butenhof, PhD Sponsor's Representative 3M Toxicology Service Medical Department J Date Steven NM. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. ! Representative Quality Asdurance Unit Hazleton Wisconsin, Inc. (6329-133. protdskz) Lay Date . 12-13-74 Date J 00315 Page 41 of 46 J PPa3ge011i6A.g Attachment 1 Scoring Seale for Acute Dermal Reactions Ervtheny 10 -- SNloingeht 32 -- HSoedveerraete Edema 03 32 T-- MNSooidngeehrtate(b(arraeilsyedpearpcperpotxibilmeatetloywe1llmm)defined Severe (raised by definiteraising) more than 1 mm) Atonia i 2017-- NSoTnigeht (slight Moderate (slow riemtpuarinrmteontnoorfmaell)asticity) 3 - Marked (no elasticity) Desquanation 10 2 -- MSNolodinegerhatte(sl(iscgahltesscaalnidngf)lakes) 3 - Marked (pronounced flaking with denuded areas) Coriacequsness 01 2 -- MNSolodinegerhatte(d(elceraetahseeryintepxltiuarbei)lity) 3 - Marked (tough and brittle) Eissuring 01:2- NSolnieght (definite Moderate (cracks cirnacdkesrniisn) epidermis) 3 - Marked (cracks with bleeding) J 00316 Page 42 of 46 HLAR3zLLSIESTSRShesY comma PROTOCOL TP3016.A8 Single-DofosTe-D6e0r5m4aalnAdbsTo-r6p0t5i1oni/nTRoaxbibciittsyStudy HWI 6329-133 Sponsor 3M Toxicology Service Medical Department 3P.M0.CenBtoexr,3B32l2d0g. 220-2-02 St. Paul, MN 55133-3220 Sponsor'sRepresentative John L. Butenhoff, php Contractor HazletonWisconsin, Inc 3301 Kinsman Boulevard Madison, WI 53704 ` StudyDirector Steven M. Glaza Amendment No. 1 iV This amendment modifies the following portions of theProtocol: NBT tro os tonto 10) 10 ray Effective December 23, 1994 In order to application obtain a measurable in Groups 2, 3, and amount of test material (T-6054) for sterile water for injection and a4,ppltiheed taetstamactoemrmioanldowsiell vobleumdeiluted with Maonddi#f2y) tthoe fionldliocwaitnegthtewsoe scehcatnigoenss. of the protocol (protocol amendomfen0t1 imtle/mksg.#1 Add the following shaded additions to this section: Dose Level Number of Anim s ) reste entosses 4321 (Control) DisTt-Til66l00e55d44water 7-6054 0o1.*.281;28% 333 333 5 T-6051 12.86% 3 3 * 3 3 * ** To be administered mTaotebreiaaldmin(fiasbrtiecr)ed at as a a |dose volume of 2.0 ml/kg 10.0-cm x 10.0-cm piece of test 00317 Page 43 of 46 | ' Anendnent No. 1 PHHaIge 62329-133 2 APtdhanesitnf.iosltlroawtiinGgoonea,rsisDtehenelteattlehirtDdheesainfgdonurfto8.hurtsDheonstseeennActdeeanicnieinssttrthaiotsitohSniessstsro(3no) troiosnisethen add sd2Tot0h1seeernicllfo/enoktrgwr.oaeltaecrhmatafenoririnaaillnjei(ncGtriGoorunopupa1ns)dw2,ialplp3,libeeadndaapt4plawiielddlosuebnedvieldujjtulemedes.eodrwTihceh Effective December 28, 1994 miySatetteheemirrsciilla#eel1 wianabntadsetehrde#2f)oto.nersttiWhnomejdieixcffttayucirtoentshtehwaiftlfolorlslGtroreeworpiuilnplagsecew2,adtie3s,rtilaflnoderd4iwnaj(teseceetrioparnsotwteihcelt]coonmytaernsoihlceertrhe indicate this change. thres Sections of the peatarey ry 3. wPiatghe 4th.e6.folCoTnotwrionlg?Materials A, Identification. Replace distilled water Sterile Water for Injection 4. Ptaegmepte.ratu6.reCwointthroltheMatfeorlilaolw:ingD.: Storase Conditions. Replace Room Refrigerated 5 Hpoadsiefy..T.he btaebrlieneinntatlhisDesseicatni:on8,witDhosetheAdnfionliTsotwriantgiosnh:ad(u1f)ThTeessrte:Groups --Srow Test Material _D(omsee/Lkeav)el HNalues omfbAFnieemmarlas 21 (Control) 3 SET water T-6054 3 oOr.28vs 33 Lagos 3 33 3 5 TT--66005514 ["EX 3 33 **+ TToo bbee aaddmmiinniisstteerreedd material (fabric) aast aa d1o0s.e0-cvnaluxne10o.0f-c2n.0pnieicekof test **4 To be adninistered at a dose volume of .01 nl/kg ) 00318 Page 44 of 46 } Amendment No. 1 PROTOCOL APPROVAL HWI 6329-133 Page 3 hn 2. Buloclrty John L. Butenhoff, PhD Sponsor's Representative 3M Toxicology Service MedicalDepartment. 1-19-75 Date j Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. c. Representat ile Quality Assdrance Unit Hazleton Wisconsin, Inc. (6329-Al1.3ds3k2.) EE Date' Date [42 G5 0031 - Page 45 of 46 @ WISCONSIN yo RRBISSINETAD aE + CORNING Corea PROTOCOL TP3016.A8 Single-DosTe-6D0e5r4maalndAbTs-o6r0p5t1ioinn/TRoaxbibciittsy Study of HHI 6329-133 Susnsern 33MMMeCTdeonixtcieacrlo,loDBgelpydagrS.tenrev2ni2tc0e-26-02 S2t..0. paBuolx, 3M3N22055133-3220 Soonsor's Representative J-oh_n-- L. Butenhoff, PhD Contractor H3Ha3a0dz1ilseoKtnio,nnsmNWIainscoB5n3os7ui0l4ne,varIdnc. Study Director Steven H. Glaza Anendnent. No. 2 This amendnent modifies the following portions of the protocol: ) Effective January 24, 1995 2SsO%oeFrcgtbtmiiheolesnesrwecioqolfullelstethbceeteopdfirnowcttilholuecldoeSldbpeowtnosidotorhci,unmctelhtnuehdteeesdwaemtpihilegnsehettshseahdiodpfrinateutniitods.nasstu.ae.HsodciTofhlyelsescttheewdeifooarnnitdsowtinhneegd voluse 1+ Ytfohadeieffy8,ol.lt7oh.ewinEsgexcpuoennrddiemresneitnnateledncDeaodsidioinnt;itohne0:. FiPrasttholpoagrya:grap(3)h SofamptlhiesCocleleccttionm.ith cdtioehnlreelmeawFclhitoresldat,eppmllawiilecvieeargt,hiaeondndbilfse(ie,vtnoeallauentfernoaoempncplraryloolpxsdiiemaetandteiermnailin1sn-,eecdancahnxdg1rb-oocutnph swkeiicldtnlieoynbse. offrotmhe -I2m0meCdi4a1t0elCy. placed in a freezer set to maintfaoifnbialet)empenrgature of 2 PYtaohgdeeiffyo5l.l7fo.hweinfsgexcpuoennrddienresnletinantleedncDeaesdidiainnt:itohne0.: sePcaotnhdolopgayr:agra(3)phSoafmpltehisColfleeccttiioonn.ith sThhieppseadmpltoesthaendpetrhseoinr cloirstreedspoinndiSnegctiwoenia7h.ts. (o4r).vo(cl)uses will be express vl 00320 ENN Le Kn maoison we asa Page 46 of 46 ) Amendment No. 2 3. Page 9, 8. Report. Add the following to this section: Individual animal tissue weights and bile volumes PHNaIge632 29-133 PROTOCOL AMENDMENT APPROVAL Pl 2. Ructnczy 2/5 fo John L. Butenhoff, PhD Sponsor's Representative Date 3M Toxicology Service Medical Department . Acuts Toxicology Steven M. Glaza Study Director ) Hazleton Wisconsin, Inc. Hed Representatije Quality Assdrance Unit HazletonWisconsin, Inc. (6329-133.Am2.dsk2) Date 2-9 2.795 Date ) 00321 9.1.2 Analytical protocol AMDT-013195.1 000322 3M EnvironmentalLaboratory Protocol - Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T-6051 and T-6054 in Rabbits In-Vivo Study Reference Number: HWI#6329-133 Study Number: AMDT-013195.1 `Test Substance: FC-129 (T-6051 andT-6054) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of TestingFacility: 933M5 EBnuvsihroAnvmeennuteal Technology and Services St. Paul, MN 55106 Proposed Proposed CInoimtpilateitoinoDnaDtaet:eJ:ulAyu2g5u,s1t92955, 1995 Method Numbers and Revisions: AMDT-M-1-0, Thermal Extraction of Fluoride by Meansof a Modified AMDT-M-2-0, Dohrmann DX2000 Organic HalideAnalyzer-Liver TFolnuoArniadelyMzeearsurement by Means of an Orion EA940Expandable AMDT-M-4-0, AMDT-M-5-0, Extraction of Fluorochemicals from Rabbit Liver Analysis of Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry AMDT-M-8-0, Analysis with Ton of Fluoride Using the Selective Electrode Skalar Segmented FlowAnalyzer Author: James D. Johnson Approved By: Ane Jdtmudeys DDi.rktoJr}inson 1/30/75 Date 600323 a2 SJpoohnnsBourtReenphroefsfe,ntPahtDive Date 1 0_PURPOSE TTflh-ui6os0r5is4nt)eu-dicysonditseardimenasilinlggynceaodbmstpooropburendod.vsidT(ehbieiontafnroaarlnmysatftioicroamnlataaissoptneocptwrohofeduttchhteissr)sFitCnu-dt1yh2ei9sti(tsoTs-du6ee0ta5en1rdmainnd of rabbits at various times post dose dermal application of FC.129, socum 2.0 TEST MATERIALS 2.1 Tes2t.,1C1onAtnraolly,tiacnald RReeffeerreennccee Substances Substance: aFnCd-9M5a,trloitce1s61 equivalent. or 171. They are 2.1.2 2.1.3 AAnnaallyyttiiccaall RCoenfterroelncSeuMbasttrainxc: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.1is4i-osner(2u.m1).1), grocery store (2.1.2, 2.1.4- ad2e.n3rdmN6aulcmoanbptperlorilocafatniiTomenaslstist.aenaTdnidsCsobuineltes.raonAldnSafallymuspiildssesoi:nfctTlhiuesdsseeuletisivseasru,nedssefwrliuulmild,bfcereloaltmutl2ha4er dftrieasstcetraionotnii,mganls the Study Director. of 24 the HIdWenItiafniicmaatlioindeonftiTfeisctataionndnCuomnbterrolwhSiacmhplceonss:isTthseosfaamlpeltetesr are identified and five digit using number, plus the tissue identity and day identity (serum). 2:5 Purity and Strength of Reference Substance: To be determined by Sponsor. 2.6 Stability of Reference Substance: To be determined by Sponsor. A22M.07DTS-tSo1r-0a1g0Ce-(02C..o1n.d2i,t2.i1o.n4s).foTreTstesatnMdaCtoenrtiraolls:saRmopolemstweimlplebreatruerceei(v2e.1d.1a)c,cording to 2G8LPDiRsepgousliattiioonnofforSptehecitmiemnesp:erBiioodlorgeiqcuailretdissfouresstaunddiefsluliodnsgweirllthbaen r2e8tadianyesd.per 2at9tirSe.afUetsye Pcraeuctaiountiwohnse:n RheafnedrlitnogakpnpirvoepsrifaotrecMutStDinSg.thWeesaamrpalpepsr.opriate laboratory 00324 2 3.0 EXPERIMEN' AL - Overview aaTnnhaaellyyttsiiiscssaulfeostrefsftrlsouocmraiannneibmecaoplmesprofdouornsmededsd..as ATdtehsetchresicdbrieesdecnr(ieHntWgiIof#no6ro3ff2lt9uh-oe1riS3dt3eu)d,inyalDriievreaervcatvioilraa,bclaoemsbefuroirsetsioofn f(lsueoerMineethiondtsh-e-nleivxetr.sTecoticoonn)fiisrtmhethaeppirdoepntriitayteofanafllyusoirsinteo-pcroensteanitnidnegficnoimtipvoeudnadtas for psrpeescetnrtomientlriyvemra(yif baenysealte2ct8eddaayss)onaendofstehreuamnaaltyvtaircailoutsecihntneirqvuaelss,eemlpelcotyreosdp.raNyomtaaslsl ocofltlheectteidsstuoesmeaentd tfhleuiodbsjaecmtpilveess woifllthbeesatnuadlyyziendt.heWohpeinniosnufoffictiheentSdtautdayhDaisrebceteonr, analysis will cease. 40EXPERIMENTAL-Methods 4.1 Liver and Serum screening methods: (attached) D4.o1h.1rmAaMnDnTD-XM2-010-00,OrTghaenrimcaHlaElxitdreacAtniaolnyzoefr-FlLuiovreirde by Means ofa Modified 4E.x1p.a2nAdaMbDlTe-IMo-n2A-n0a,lyFzleuroride Measurement by Means of an Orion EA940 4.1.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver 4U.s1i.n4gAEMlDecTt-roMs-p5ra-y0M,aAsnsalSypseicstroofmReatbrbyit Liver Extract for Fluorochemicals 4A.n1a.l5yzAeMrDwTi-tMh-I8o-n0S,eleAcntailvyesiEsloecftrFoldueoride Using the Skalar Segmented Flow 5.0 DATA ANALYSIS t5fi.ls1usouDreiadtoerapfeRlrueiptdoi.rsstSuiteantgoi:rstfilDcuasitdu,aseowdri,lalastbFetChr-ee9dp5iosr(ctereledetciatosrnoaosfpctrohanyecemnSattsruasdtyisopDneicr(tewrcetoiomgreh,ttrm/ywa)eyipgeihrntcu)lniuotdfeof rdeegvrieastsiioonns aonfacloynsicseontfrsaetirounms fcoorncdeinftfreraetnitondsosweitghrotuipmse.anIdf naevceersasgaersy,ansdimsptlaendard statistical tests such as Student'st test may be applied to determine statistical difference. 600325 S 6D .0 MAA INTT ENAA NCE OA FRN AW DRECOR~ D~S 6.1 Raw Data and Records: Raw data, approved protocol, appropriate specimens, approved final report, and electronic data will be maintained in the AMDT archives. 1~ 0REFERENCES ~~ 7.1 AMDT-S-10-0, Sample Tracking System 8.0 ATTACHMENTS 8.1 AMDT-M-1-0, Thermal Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver 8.2 AMDT-M-2-0, Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer 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 8.5 AMDT-M-8-0, Analysis of Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 00326 4 3M Environmental Laboratory Method Thermal ExtractionofFluoride by Means ofa Modified Dohrmann DX2000 Organic Halide Analyzer - Liver Method Identification Number: AMDT-M-1 Revision Number: 0 Adoption Date: /o- #-1< Revision Date: None Author: Rich Youngblom Approved by: os Zoup Lead, fof3/7< Dale A Quality Assurance 2D-ate 7 Software: MS Word 5.1a Affected Documents: AMDT-M-2 Fluoride Measurement by Meansof an Orion EA940 AMDT-EExpPa-n3daRboluetfionneAMnaailnytzeenrance ofa Modified Dohrmann DX2000 Organic Halide Analyzer 00327 1 0 SCOPE, APPLICABLE COMPOUNDS, AND MATRICES f1l.1uoSricdoepef:roTmhivsarmieotushomadtriiscfeosr. thTehoepefrlautoiroidneoisfatyDpiocharllmyacnonllDecXt2e0d0in0TwIhSeAnBitsoisluutsieodn tfoorexatnraalcytsis with an fon selective electrode. 11.23 MAaptprliicceasb:leBiCoolomgpiocaulntdisss:ueFs,lupoarrtoicchuelmarilcyallisveorr. other fluorinated compounds. 2.0 KEYWORDS D2.X12F0l0u0or,idfel,uofrloucorhienmei,ceaxltsr.action, pyrolysis, ionization, ion selective electrode,Dohrmann,halide, 3.0 PRECAUTIONS 3.1 3.2 Glassware Glassware and exhaust gases can is fragile, broken glass be extremely hot. may cause injuries. 33..43 SPorlesvseunrtibzeadsegdasseasm,pplreospemracyofmlparshe,ssmeadygnaesehdantodlailnlgowprtacthicteesodrmreqyuidreodw.n before starting run. 3.5 Potential equipment. biohazards due to the biological matrices. Use appropriate personal protective 4.0 SUPPLIES AND MATERIALS 4.1 4.2 Compressed Compressed Oxygen, Helium, Hydrocarbon free, High Purity Grade, regulated regulated to to 30 45 PSL PSI. 44..43 QQuuaarrttzz ggllaassss csoammbpulsetbiooanttwuibteh, RTeelfilaonnceTMGtluabsisng,G-D9o4h0r5m-a0n1n2 8o9r0e-qu0i9v7aloernte.quivalent 4.5 4.6 OSraimopnle94c0ol9l9e9ctTiootnavliaIlosn,iHc DSPtrEe.ngth Adjustment Buffer (TISAB II) or equivalent. 4.7 Milli-QTM water 4.8 4.9 PAocltyisvtaytreednCehpairpceotatle,s.E. Merck 2005 or equivalent. 44..1110 MHiasmcielltloanneSoyursinlgaeboorrateoqruyivgalleansts.ware 50 .0 EQUIPMENT 00 5.1 5.2 IRoBsMemcoomupnattiDbolher3m8a6nnorD4X3260c0o0mpOurtgearn.ic Halide Analyzer, modified for fluoride extraction. 5.3 5.4 DX2000 software, Excel Spreadsheet, version version 1.00, modified 5.0 or greater for fluoride extraction. 6.0 INTERFERENCES e---- m6.a1ySvaamrpylferosimzemaitsrliixmittoedmattoriaxp.proximately 150 mg, depending on sample moisture content. This 600328 2 21.0 SAMPLE HANDLING 7.1 Samples are not to be handled with bare hands. sample. Use forceps or probe to transfer tissues. Fluoride may leach from the skin to the 7.2 Samples of liver are cut from frozen liver and placed ina tared and labeled weigh boat. Usea. clean scalpel and cutting board. The cutting board and scalpel should be cleaned with water, methanol, or methanol-water solution after each liver is cut. 8.0 CALIBRATION AND STANDARDIZATION 8.1 Preparation of Calibration Standards 8.1.1 The standards required for each Refer to protocol for that project. project will need to be appropriate for that individualproject. 8.1.2 Typically 50-500 ppm FC-95 in methanol standards are used. 8.1.3 For rabbit liver studies, usebeef liver as the matrix. 150 mg) and weigh it in a labeled and tared weigh boat. Cut a piece of frozenbeef liver (100- 8.2 Calibration - Overview Tcuhrevenoirs mraelquciarleidbrtahteiponroicsetdhuerfelluiosrtieddebceulrovwe (AMDT-M-2). is used. However, if an optional spiked liver 8.2.1 A calibration curve for the DX2000 is generated by and combusting them using the same methods and matrix spiking samples with type as the samples to known standards be tested. 8.2.2 Typically, spiked. three replicatesofeach standard and five concentrationsof standards will be 8.2.3 Standard curve will be plotted as Mass Spiked F (ug) on the x-axis and Standard Mass Recovered F (ug) on the y-axis. Generate a regression curve and calculate the. equation for the line and the r2 value. 8.2.4 Mass Spiked F (ug) = (Amount spiked in mL) x ( Conc. of standard in ppm) x (0.6004)* 8.2.5 *FC-95 is 60.04%Ftherefore 0.6004 is the Standard Mass Recovered F (ug) = (TISAB volume factor used to convert FC-95 to F in mL) x (Orion reading in ppm) 8.3 Calibration - Procedure 8.3.1 Start Up 83.1.1 Run 2 or more Clean Cycles when starting instrument each day. More clean cycles may be used if the previous samples contained high concentrations of fluoride. 8.3.2 Blanks 83.2.1 Prepare sample using the same methods and tyofmpatreix as the test sample. 8.3.2.2 For rabbit swdies, use beef liver as the matrix. Prepare at least 3 samples ofbeef liver (100 - 150 mg) for blanks. 8.3.2.3 analyze Put sample in Dohrmann boat. Combust sample according to method AMDT-M-2 each sample as described in section 9.0 for the ion selective electrode analysis. and 00329 3 8b.e3f.o2r.e4Fproorcereadbibnigt wsitutdhietsh,e tchaelimberatteironr.eaBdiunmg sfoarmpalbelsanunktislatmhpilselismhiotuilsdrebaec0h.e0d3, oprpumntoilr ilnowtheer judgement of the operator the reading is stable with respect to historical readings (previous 48 hours). 8.3.2.5 For non-rabbit before proceeding with stthuediceasl,ibtrhaetiboln.ank readings should reach a predetermined ion concentration 8.3.2.6 It may be necessatroy mix approximately 50 mg of charcoal with the sample to aid combustion. 8.3.3 Standard Curve 83.3.1 Weigh out at least 15 matrix samples (5 standards with 3 replicates each) in tared and s8lt3aub.ed3ly.e2ddatwPae.liagcShetobwroeeaittgsh.heemFdaboterreirfxalbsbiiavtmeprsltsueadsimeopsnl,ewdreiynigDihcoeh1or0rm0ia-c1ne5np0asmcakgmsbpeltoeefkbeloaietvp.etrhseammpflreos.zenRuenctiolrdusweed.ights in 83.3.3 Start with the lowest standard concentration. Using a Hamilton syringe, ejecta fixed quantityofthe standard on or in the matrix. For rabbit studies, use 4uL ofstandard and eject iton orin thebeef liver. m8.a3y.3b.4e uAsteldeaasttt3herdeipslcicraetteisonsohfotuhled banealuyssetd.for the lowest standard concentration; more replicates 8.3.3.5 Combust the sample as described in section 9.3 and analyze according to AMDT-M-2. 8.3.3.6 Run all 15 standards.If one replicate is significantly different from the other two roelpdlirceaptleisc,atreuannadnotthhaterthseamnpelwerfeoprlitchaatteswtianldabred.useIndditocactaelciunldaatteatthheartetghreesnseiwonrceuprlviec.ate replaces the 8.3.3.7 When all standards have been run, calculate the 2. r2 must be at least 0.95. Ifitis not at least 0.95, consult with supervisor. 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 of the analyst. matrix is 8.4 Storage Conditions for Standards 8.4.1 Storage requirements for 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. 9.0 PROCEDURES 9.1 Typical Operating Conditions: 9.1.1 Combustion tube temperature = 950C. 9.1.2 Oxygen and Helium flow = 50 cc/minute. 99..11..34 VBaapkoeritziamtei=on3/0Dr0ysiencgontdism.e = 240 seconds. 9.2 Start Up Procedure: 9.2.1If the program is not started, start the EOX program on the PC. 9.2.2 Open the SYSTEM SETUP window. 9.2.3 Put the fumace module and the cell in the READY mode. 9.2.4 Close the SYSTEM SETUP window. 4 00330 9f:o2u.n5dWihnetnhetCheEoLvLenChHaEsCreKacmheendut.he READY temperature, run the CLEAN BOAT program 9.2.6 See AMDT-EP-3 for details of the Dohrmann software. 99..33.1SaOmppelnetEhxetSraAcMtiPoLnEPrHoAceTdCurHe:and place the sample in the BOAT. It may be necessary to mshioxuladpparlosxoibmeatmeilxyeSdOinmwghoiflechesatracbolailshwiintghtthheebsaasemlpilneetaonadidwhceomnbguesnteiroant.inIgftthhiesstisanddoanred, ccuhravrec,oal 9.32 9.3.3 ACdlodseapSprAopMrPiaLtEe vHoAluTmCeHo.f TISAB solution or 1:1 TISAB:Milli-QTM water mixture to a labeled 2.0 mL soafmpl1: eTcIoSlAleBc:tMioinlvliial-.QTMTwyapitcearllmyix0t.u6rme.L to 15 mL are used. For rabbit studies, use 1.0 or 9in.c3h.e4s.PlGaacseetsherevliealasseod that the during ptyiproolfytsihse mCuOsMt BbuUbSblTeItOhrNouTgUhBthEe iTsIiSnAtBh.e TISAB at least 0.25 99..33..56 WRhunenthteheEOEXO-XSpOrLoIgDraSmpirsofgirniasmhefdo,urnedmionvethtehRe UcoNllemcetnioun.vialfromthe combustion tube. 9.3.7 10 the If undiluted TISAB TISAB to make 1:1 was used to collect TISAB:Milli-QTM. the sample, add an equal volume of Milli-QTM water r9.e3m.o8vReinasneytThIeSeAndBofretmhaeincionmgbuosnttihoenttuubeb.e with Milli-QTM water and wipe with a KIMWIPE to 9w.i3t.h9a Ocpotetnonthteipspaemdpalpeplhiactacthoraonrd vraecmuouvmeeadnoyutr.emIatimnainygbeasnhefcreosmstahertoybosact.rapApsahrtciaclnesbeofrfetmhoe.ved bbootattotomawiidthinathsepaCtluelaanorCyoctlhee.r similar device. A dropofMilli-QTM water may be added to the 9.3.10 Close the hatch. 9.3.11 9.3.12 SRaumnplteheisCrLeEadAyNfoBr OanAalTyspirsobgryaimo.n selective electrode (AMDT-M-2). 9.4 Sample Calculations 9.4.1 9.4.2 USasmeptlhee sMtaasnsdaRrdeccouvrevreetdo cFal(cuugl)at=e(tTheISsaAmBplvoelvianlume.L) x(Orionreadinginppm - intercept) (Slope) 10.0 VALIDATION 1100..111QuDaailliytyStCaornttrUoplCheck Samples: Once the standard curve is established, each day of caonnaclyesnitsraitsisotnarstpeidkebdy saanmaplylzeisngusQedCtsoagmepnleersa.teTthhee sQtCandsaarmdplcuersvea.re Etoacbhectohnecsenatmreataisonthmeulsotwebset dreopnleiciantetriispnlioctatneecuenslseasrsy.the first two replicates are within 20%ofthe standard curve, then a third F1l0u.2oriPdreecNiostieobnoaonksd 2A.c3,cuarnadc5y.: PrSeeceismieotnhaondddaecvceulroapcmyevnatriaensalwyhseisnaanndalsyazmipnlgesaanmapllyessiosfidnifferent matrices and different reference compounds. 10.3 Other Validation Parameters: NA 000331 5 11.0 DATA ANALYSIS 1L1 Calculations 11.11 11.1.2 TFoorctahlecusltaatnedtahred fcluurovrei,deusceonrtergarcetsisoinoninatnhaelyssaimspilne,Exsceeel,mevterhsoidonA5M.D0To-rMgr-e2at.er. 11.2 Analyzing the Data a1r1e.2w.i1th1i2n m2u0s%tobfeeaatclheasotth0e.r9a5nodr tghreeaotuetrl,ie"rOuits lgireerast"ermtahyan b2e0e0x%colfutdehdeiaftvewroagoefotfhtehtohsreeetrweoploicrates lneostsedthiannt5he0F%ionfalthReepaovretraagleonogfwtihtohsethtewor.easAonnyitswuachsocuotnlsiiedresrsehdoaunldoubtleipeori.ntedoutin the data and 1A 20 TTAC ~H ~~ MENT 000S 000 None 13.0 REFERENCES 13.1 349, Rosemount revision B, Dohrmann DX2000 December 1993) Organic Halide Analyzer Operator's Manual (Manual 915- A1n3.a2lyzAeMrDT-M-2 Fluoride Measurement by Meansofan Orion EA940 Expandable Ton 13.3 AMDT-EP-3 Analyzer Routine Maintenance ofa Modified Dohrmann DX2000 Organic Halide 14.0 REVISIONS Revision Number ReasonforChange. Revision Date 00332 s 3M Environmental Laboratory Method Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer Method Identification Number: AMDT-M-2 Revision Number: 0 --- Author: Rich Youngblom Approved By: Grod] Leader Adoption Date: /0-4-45" Revision Date: None LL/2s Date , A R 5 AL Quality Assurance Lo-#7F5 Date Software: MS Word 5.12 DAfofhercmtaednnDoDcXu2m0e0n0tsO:rgAanMiDcTH-alMi-d1e ATnhaleyrzmearl Extraction of Fluoride by Means ofaModified 00333 1 . L0SCOPE,APPLICABLECOMPOUNDS, ANDMATRICES 1.1 SCOPE: fon Analyzer. This method is for the calibration and operation ofan Orion EA940 Expandable 1.2 APPLICABLE COMPOUNDS: Fluoride. p1H3 oAfP6P.0L. 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. 4.0 SUPPLIES AND MATERIALS 44..12 OOrriioonn M9o4d0e9l9990To0t0a0l1TeolneicctSrtordeenfgitlhlinAgdsjoulsuttmieonnt(BAugfCfle)roIrIe(qTuIivSaAlBeInIt,) or equivalent, 4.3 4.4 Orion 940907 100 Milli-QTM water or ppm fluoride equivalent. standard or equivalent. 44..56 MLaagbnetitsisucess.tir bars. 44..87 SPlaamsptilce1c0o0llmecLtiovnolvuimase.tric flasks. 44..910PoMliysscteylrleanneeopiupsetltaebso.ratory glassware. 5.0_ EQUIPMENT 55..12 OOrriioonn MMooddeell 9E6A0994000 ESxolpiadndStaabtleeCIoomnbiAnnaatliyoznerFolruoerqiudievaelleenctt.rode or equivalent, 5.3 Magnetic Stir Plate. 55.:54 OIrBiMoncRomSp2a3t2ibilnete3r8f6aceorca4b8l6ec(oomnplyutneered(eodnliyfnuesiendgedOirfiuosni3nEg Osorfitownar3e)E. software). 5.6 Microsoft Excel 5.0 (only needed ifusing Orion 3E software). 6.0 INTERFERENCES 6Q.1mItwiastrerecwiolmlmgeenndereadlltyhabtrithneg psaHmpbeleattoorpnHeofa6r.06..0. A 1:1 mixture ofTISAB and sample/Milli- a6t.2rSoaommptleempteermapteurraetuarsethmeasytaenffdeacrtdsflwueorreidewhmeenastuhreemmeentte.r It is recommended was calibrated. that the sample be 6.3 The rate the samples are stired at should be consistent with the rate the standards were stirred. 00334 2 6u.n4deArirelbeuctbrboldees. trapped under electrode can give erroneous readings. Make sure no air is trapped 7.1 No special handling necessary. 88..11.1PrMeepaasruarteio5n0 mofLCoafliTbIraStAiBon[TSitnatnod5ar1d0s0 mL plastic volumetric flasks. 88..11..32 PLiapbeetlteth0e.0f5l,as0k.s1,a50.05.,051,.0,0.a1n,d0.15.,51m.0L, aonfd110.05ppppmmfFl-u,oarildoengstwaintdhartdheindtaotethaenadpyproouprriinaitteilalys. labeled flasks. 8.1.4 Add 8.1.5Shake atphperofxliamsaktsetloym3i0xmthLeosfolMuitliolnis.-QTMwaterto each flask. 8t.h1e.f6lEaslkismionvaetretahierabiurbbbulbeblsefs.rom the flasks by tipping the flasks on their sidesandrolling theairin 88..11..87IBnrvienrgtatnhde vsohlaukmeethien ftlhaesfklsafsokrs tuhpetfointahlem1ix0i0nmg.L mark with Milli-QTM water, 8.1.9 Record standards in Standards Log Book. 88..22.1CIaflniebcreastsiaorny, remove tape from electrode filling hole. 8.2.2 8.2.3 EIjnevecrttapfreobwedrtoopwsetoftofipllsianlg.solution from bottomof electrode to wet lower seal. 88..22..54 TFihlel tmheeteelrecatnrdodtehewiFt-helfielcltirnogdseolaurteiotny.pically calibrated by direct measurement with no blank cthoememcatniuofna,ctuusrienrg'sstiannsdtarrucdtsiownist.h concentrations of 0.05, 0.1, 0.5, 1.0, and 1.5 ppm Fo, following 8.2.6 8.2.7 Record the slope in Clean the electrode the appropriate by rinsing with log book. Milli-QTTM water and wiping the sides down with lab tissues. 88..33.1SCtaolriabgreatCioonndsittanidoanrsdsfoarreStstaonrdedaradtsroom temperature. 9.0 PROCEDURES 99..11.1CaTlhiebrsaatmipolneatnodbeMemaesausurreemdennte,edSsttaonbdearmdixmeedthwiotdh:TISAB using the proportions r9.e1c.2omPmlaecnedaedstbiybatrheinTIthSeAsBammpalneufaancdtuprlearc.e the sample on the stir plate. e9l.e1.c3trAodleloiswitnhseerstaedm,plmeaktoemsiuxreftohrearefaerwesneocoainrdsbubbebfloersetirnaspepretdinugntdheeretlheecterloedcet.rodWe.hen the s9.a1m.e4 Trahtee assamtphelecaslhiobrualtdiobnestthaendsaradmse. 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. 00335 3 re 9.2 Calibration And Measurement, Using Orion 3E Software: TeapY AVAILABLE 99.22..11.1CaFloilblroawtisotnep:s 8.2.1 108.2.4 9.2.1.2 Press Function Key #8 (FS). c9.o2n.c1e.n3trTahteiocnoomfputhteesrtsancdraeredns,wialnldaswkhyoeuttoohrcenonorftiarmbltahnekniusmtboebreoifnsctlaunddeadridnstthoe cbaeliubsreadt,ion. 9M.a2k.1e.4aPnlyacneectehsesaelreycctrhoadnegeinstthoetihersinsftoarnmdaatridononprtehseesnttredplaantde aclnidckcloinckCoOnNCTOINNTUIE,NUE. s9t.a2b.i1l.i5zeOdb,seprrevsestAheCrCeEadPiTngsRoEnADthIeNgGra.phic display on the computer. When the readings have 99..22..11..76 RAfetpeeratthsetfeipna9l.2s.t1a.n4daarndd, 9t.h2e.c1.o5mpfuorttehrewriellmadiinspilnagystthaendsalrodpse.of the curve, as well as the ainntdecrlciecptkoanndCcOorNreTlIatNiUonE..ReTchoercdalthiebrsaltoipoen,dianttaericsepatu,toamnadticcoarlrleylactoiopnieidn ttoheC:a\ppOrroipormiDaatteal\oCgalbiobo.kxt. 99..22..22.1DaStelaeSctpreeitahdesrhNeeEt:W or OPEN from the FILE menu to open a new or existing spreadsheet t9o.2s.to2r2eRdeactaoridn.the nameofthe spreadsheet used in the appropriate log book. 99..22.33.1FlFuoolrliodwe sMteeapssu9.r2e.m1etnhtr:ough 9.2.4 99..2233..23 ECnltiecrk tohne tnhaemNeoEfWthSeAsMamPpLleEibnutthteonappropriate place on the screen. a9p.p2r3o.p4riWahteelnogthbeoorke.adings have stabilized, click on the RECORD buttonand write the resultinthe 10 0.0 vALmpATIO0 N 00 10.1 Quality Control: 10.2 Precision and Accuracy p10p.m3 fOltuohreirdeVuaplitdoaatisoantuPraatreadmseotleurtsionoAcfcfolrudoirindget.o Reference 13.2, the raofndetgecteion is 0.02 11.0 DATAANALYSIS 11.1 Caleulations None necessary. 112 Analyzing the Data None necessary. 12.0 ATTACHMENTS None 13.0 REFERENCES ~~~ 000000 _ -------- _ 00336 4 13.1 Orion Model EA940 Expandable lon Analyzer Instruction Manual, Orion Research Incorporated, 1991. 13.2 Orion Model 960900 Solid Research Incorporated, 1991. State Combination Fluoride Electrode Instruction Manual, Orion 40 REVISIONS 0000 RNevuimsbieorn ReafosrChoannge Due Revision 00337 s Method Extraction of Fluorochemicals from Rabbit Livers SOP Identification Number: AMDT-M-4 Adoption Date: /o-2/-55 Revision Number: 0 Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: up Leader Jo--3/-25 Date 4Lf Bhd, Quality Assurance lias Dae SAfoffetcwtaerde:DoMcuSmeWnotrsd:, M6-.05, AnalysisofRabbit ExftorFrluoraochcemictals Using Electrospray Mass Spectroscopy. 000338 , Scns sud oc 1.1 Scope: This method is for the extraction offluorochemicals from rabbit livers, Ethyl acetate is used to extract fluorochemicals from the liversforanalysis by 11..32 MAaptprliicceasb:leRaCbobmiptoLuivnedrss.: Fluorochemicals or other fluorinated compounds. 2.1 cFolmupooruocnhdesm,iceaxltsr,acrtaibobni.t livers, electrospray mass spectrometer, fluorinated 4.1 Sup0plieSsyringe, capableof measuring 100 uL. 4.1.2 Eppendorf typeordisposable pipets 44..11..43 PGllaosvtiecsgrinding tubes 4.1.5 Plastic centrifuge tubes, 15 mL 4.1.6 Labels 4.1.7 Nitrogen 4.1.8 Timer 44..11..910 AFnialtleyrtsi,cTailtpainpensy:lognlassysrivnogleumfieltteircs,p0i.p2etp.m. 44..11..1121 DCirsipmopsacbalpeapultaosvtiiacls3.cc syringes. 4.2 4R.e2a.g1enAtqsueous Ammonium ppm aqueous solution Aocfeatmatmeo(nAildurmicahc)e,taatpeprboyx.ad2d5i0ngpp2m5:0 Pmrgepaarmemao n2i50u0m a`wcaettear.teDtiolu2t1e0t0hims Lsolvuotliuomnet1r:i1c0fflaosrak a2n5d0dpilpumte stooluvtoiloun.me withMilli-Q 4.2.2 S(oNd&i,uCmOca/rNbHonCaOte)/S0o2d5iuMm:BiWceairgbhon2at6e.Bgufoffesro(dJ.iTu.mBackaerrb)o,nate (N8,C0,) barnidng21.t0o vgoloufmseodwiituhm Mbiilclair-bQonwaatteer(.NaHCO) into a 1 L volumetric flask and 44..22..34 EDtihlyulteAcaceettaotneitrile solution, dilute acetonitrile 1:1 with Milli-Q water. 4.2.5 Methanol 4.2.6 Milli-Q water 4.2.7 4.2.8 F1CH-,95JH(23HM2SHpe-cpiearltfyluCohreomocitcaanlesDuilvfiosniiocn)acid(Aldrich) 00339 2 5.1 Ultra-Turrax T25 Grinder for grinding liver samples. 5.2 Vortex mixer 5.3 Cenuifuge 55..54 ASnhaalkyetrical Evaporator 7.1 Tpheerfroarbmbeidt.livers are received frozen,andmustbekeptfrozen untiltheextractiions 8.1 Preparation of Internal Standards 8.1.1 Prepare an internal standardof approximately 12 ppm 1H,1H,2H,2H- 8.1.2 pWeerihugohroaotlcetaasnte0s.u1lpghoofni1cH,ac1iHd,t2oH,b2eHa-dpdeerdfltuooeraocohctlainveers`uslapmhpolen.ic acid into a 8.1.3 B1r0i0nmgLifvuoplutmoevtorliucmfelawsikt.hRemectohradnothle, atchtiusails`wtehiegshtto.ck standard, 8.1.4 Toa250 mLvolumetric flask, add 3 mLsof the stock standard andbringto `volume with Milli-Q water. Calculate the actual concentration of the standard. sacptuahlomgtpiereflauocroictdane- X 3mL = 0.1L 250 mL. actual concentration, ppm 8.2 Prepare FC-95 Anion Standards 8.2.1 Prepare FC-95 standards for the standard curve. 8.2.2 RWeeciogrhd atphperaocxtiumalatweeliygh1t0.0 mg of FC-95 into a 100mL volumetric flask. 8.2.3 Bring up to volume with dillte acetonitrile. 8.2.4 aaDipslpourltouextiitmohanetoseofllyaupt1ipo0rn0oxwp.iptm1h.0dpiDlipulmtu.eteactehtiosnistorliulteio1n:110:1f0owaritsholduitliuotne oacfetonitrile for 8.3 P8r.e2p.5areU5.s0eBpetephfe,1L0i1.vp0epprmpHsmoolmaunotdigoen5n0t0ao tpmepabk.feo`wUosrekinfgorstSantdaanrddsarwdisth values close to 8.3.1 2W0e0igmhL4s0MgiolfliB-Qovwianteer.livGerriinntdo tao a25h0ommoLgeNnaolugsenseolbuottiolne.containing 8.3.2 Add 1 mL of the solution to a 15 mL Sociige tube. Prepare a total of eight s1umspLenadlisqoulouttsioofn tbhye ssholauktiinogn iitnbe15tmweLenceanltirqiuoftusg,e tubes. Be sure to re- 00340 5 8.3.3 Sstpainkdeasrdesveinn osftetphe9.112moLfatlhiequportoscewdiutrhet.hOenfeol1lomwiLngalaimqouuotntsserovfeswoarsktihne.g blank. (AWpoprrkoixnigmatSetaCnodnarcd.) "cAopnpcreonxtirmaattieon foinf ---- FCB-9l5T winkliver 8.4 Calculate the actual valueofthe standards: _1L7.1xofm_sgtaclnivdoearr'nd/ Tcmlehonmotgernaatet(iinpopmn) = final concoefnFtrCa-t9io5n in(plipvme)r F*1ACmv9Le5rhaiogsmeroewgpeeoinrgathtetdeosafsobfeoq4vu0iinvmealelievlneitrvseironfinsFo2Cl0-ut09im5onLpoastoafdseMstiielurlmim-isQnaletw,datbeyr.weTihgehianmgount of 8.5 8C.a5l.i1braEmtxetitrohanocdt.tUhesesptihkeesdebseteafndlairvedrsthooemsotgaebnliastheyfooulrlocwuirnvge9o.n13thte0 m9.a2s3sof this 8.5.2 Aoslfptetcehtmeraotpmeiervtfeellruy.o,raooscttaanndsaurldfocnuartveeamnaioynbaendgetnheeraitnetdemuaslinsgtarnadtairosdoafnrieonspvoenrsseuss concentration of the perfluorooctanesulfonate anion. 8.6 S8t.o6r.a1geNeCwonsdtaintdiaorndss afroerprSetpaarneddawridtsh each analysis. Standards are stored in `iscopveerrfeodrpmleadst.ic centrifuge tubes unil the analysis on the mass spectrometer 8.7 8S.t7o.r1ageBeCfonldivietrihoonmsogfeonratSetsamnadyarbdesfrozen after preparation. 9.1 pOabctakiangefdrowzietnhloitvehrersatmipslsueess.. In spent tissue, note that the liver has not been 99::32 9.4 WRUeescieograhdditthsheseesclatiimvneprglwesecdiailgrphceetltilanyntidhnecoustatourdfyefdnoaptlpeapsrbtooixocikmg.artienlydi1nggoufbeli.ver. 9-5 Pinuittiaalsl.abel on the vial with the study number, weigh, rabbit ID, date and analyst 00341 4 99..76 GmAirdniduntd2e.st5,hemunsLtaismlpwtlahete.esrP.aumtptlheeigsrainhdoermopgreobneeoiunsthseolsuatmipolnewiatnhdngorilnadrgfeorcahbuonukts.2. 99..98 ATRaMiknDseTe-ttEhhePeg-pr2ri2nodb.eeroafpfaritntaontdheclseaamnpiltewiwtihthme2t.h5amnLolsawfatetrereaucshinsgamappliep.erF.ollow 99..1110 PiCinapfpeotrtmhIaemtisLoanmipanlstoethaaen1gd5rmvionLrdticenexgnttfruobiref.1u5g(eSseetceuobAne.dMsLD.aTbe-lMt-h4e cWeonrtkrisfhuegeettfuobre dwoicthumtehnetiidnengtitchael 9.12 Srpeimkaeintihnegbseteefpsl.i)ver homogenates as described in 8.3. with the appropriate amount of FC-05 standard 99::1143 aAScpdeitdkate1em.thLe osafmpthleessoadnidubmeceafrlbiovneratheo/msoogdeinuamtbeiscawribtohna1t0e0buufLfeorfainntder1namlLstaanmdmarodn,ium 99..1165 CUsaipntghaensaanmapllyetiacanldpviopertt,exad2d05tmo L30etshecyolndacse.tate. 99..1178 oCPenunttthtreihfecuemgneitnrfiotfrhueg2es0htatooke225r5.fomirn2u0temsi,nu.ntil the layers are well separated. Set the power 9.19 9:20 mReLmgorvaedu4atmeLdsgolfastshpeipteotp.oTrrgaannsifcerltahyeeltarobealfrteosthhe15fmreLshcteunbter.ifuge tube with a 5 Blow the sample down on the analytical evaporator to near dryness with nitrogen, 99::2221 aBrpipnrgoxtihmearteemlayi3n0intgo Vortex 15 seconds. s4a0mmpilneutuepsi.n 1mL dilute acetonitrile with an analytical piper. 9.23 stThtreuadnsysafnmeuprlmtebheeirnstaaonmadfprlaeenstiho.ma acel3ntnmruimLfbuesgrye.ritnugbee.orAt2taacuthovai0al..2 pLambenlylthoentmuebsehorfiveiarl,waintdh ftihleter 99..2254 CCoamppalnedtehoAlMdDfTor-aMn-al4yswiosrbkysheeleetctarnodspartatyacmhatsospsapgeectorfossctoupdyy.notebook. 100V10A.L1 IQDuaAliTtyICOonNtrol - not applicable 10.2 10.3 OPrtehceirsiVoanliadnadtiAonccPuarraacmye-tenrots.apnpolticaapbplleicable 11.1 None 12.1 Worksheet AMDT-M4, 13.1 AMDT-EP-22 Routine Maintenance of Ultra Turrax T-25 R1ev4is0iRonEVISIONS Number ReafosrChoannge 000000000 Revision Dae 00342 5 BEST ES MRA op Worksheet AMDT-M-4 ET SaNummpbieer | approx 0555ppm | approCx5p5pm | apprCo.95pom | niTtosefaonrda, | = ER [Er[om ew] rr f r e e r e r r r E r E p P r r s ------m ---- -- f Ey t -------- -- tr tt rt tf sf a-- em oe rr rr ---- ee --eee] fo em ee ee rmar are} e me e) -------- Ee eTe T -- So --r------------] ATR L e oe l E-- e ---- e re eer POT Ser-- er --} 00343 6 3E M nvir Labo oratn orymental Method Analysis of Rabbit Liver Extract for Fluorochemicals using Electrospray Mass Spectroscopy SOP Identification Number: AMDT-M- Adoption Date: (-( -45 c e me erst -- eee Revision Number: 0 Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: up Leader 4 : Quality Assurance es Date Lh fee Dae Software: MS Word, 6.0 Affected Documents: M-4, Extraction of Fluorochemicals from Rabbit Livers 00344 1 1.1 ifSslcupoiepdresf:ofTrorhmielsduombreyotcshhionedgmliiecsaifolonsr mtuohseniinagtnoatrlhiyensigesloeofcfFterCxo-ts9rpa5rcataysnmiooafns,rsaMbsb/pieZtc=tlri4vo9em9re.toertrho.etThihentreetrainnsaaslluyessiosr standard MZ = 427, and other appropriate masses. 1.2 Applicable Compounds: Fluorochemicals or other fluorinatedcompounds. 1.3 Matrices: Rabbit Livers (samples), Beef Liver (standards), other tissues and fluids. spectrometer, rabbit livers. 3.1 Use caution with the voltage cable for the probe. When the voltage cable is plugged into the probe DO NOT TOUCH THE PROBE, there is risk ofelectrical shock. 3.2 Do not run the pump above it's capacity of 4000 psi. Ifpressure.foes over 4000 psi fsitnodp tahnedprleulgeaasnedprreepslsaucree.tTheheplpuegagkedtutbuibinnggm, aSyeebeAMplDuTg-geEdP.-T1r5oubleshoot back to 3.3 Do not run the pump to dryness. 4.1 4S.u1p.p1liNeistrogen gas regulated to 140 psi. 4.1.2 4.1.3 F1l0u0ofuiLx ocrol2u5m0nuoLrfelqatuitviaplseynrti,nge for sample injection. 4.2 Reagents 4.2.1 Dilute acetonitrile mobile phase, dilute acetonitrile 1:1 with Milli-Q water. 4.2.2 Milli-Q water, all water used in this method should be. Milli-Q water. 5.2 5.3 SIpSeCcuOraSpyhryisnigcesPAuSm3p00 Autosampler 5.4 100 uL Assembly 5.5 Autovials or capped centrifuge tubes. 6.1 There are no known interferences at this time. 7.1 uKneiielprtehaedyefxouracatneadlyssiasm.ples in capped 15 mL centrifuge tubes or in capped autovials 00345 2 8S .0CT A8.L1IBA RPrAepTN arIaOtiNoD nAoNf A CDalibrR ationD StandI ards ZATION 8.1.1 Sexetvreanctiboenefprliovceerdusrtea.nd(aSredes aAnMdDoTn-eMb-l4a,nksbeecetifolniv8e.r0)are prepared during the 8.2 8C.a2l.ib1raRtuinonthe sevenbeefliver standards twice, stating with the lowest standard 8.2.2 T0ypoibctaalilnytohneestsatnadnadradrdcuirsvreu.n after each 5to 7 samples. Choose a standard in the same rangeofconcentration as the samples. 8.3 8S.t3o.r1ageFreCsohndstiatnidoanrsds for are pSretpaanrdeadrwdisth each analysis. Standards are stored in `cpoevrefroerdmepdl.asStaicmpclenetsriafnudgesttaunbdeasrudnstialrethNeOanTalryesfirsigoenrattheed.mass spectometer is 8.4 8S.t4o.r1ageBeeCfonlidvietrihoonmsogefonratBeseemfayLibveefrroHzeonmaofgteernparetpeasration. 9.1 9In.i1t.i1al SSetetsoufptware to "Operate on", Ion Mode ES" 99..11..32 FRiellcotrhedsboalcvkeinntgcyplriensdseurrewiinththmeobiinslterpuhmaesnet.log. 9.1.4 cSeotmithnegpouutmopfttohe"Rtuinp"o.fStehte pthreobfel.oTwhteo p1r0e0s0suurLe/smhionu.lOdbbseeratve1s7d0r0optloet1s800 9.1.5 Chsii. ck the fused silica at he end ofthe probe. Use an eye piece to check for Cchuitposf.fTthheettiipposfhtohueldsibliecaflwatitwhiathcnoolujmanggceudtteedrgaensd, pIuflalntyhechsiipiscaartehrfoouugnhd10 9.1.6 tChheecapkpryoopurriantietrloegnegnthsupply. Tum on the nitrogen. There should be no onuittroofgetnheletiapk.ing around the tip of the probe. A fing mist should be coming 9.1.7 fCaarrtehefru.llCyognuniedcettthheepvroolbteagientcoabtlhee toopetnhienpgr.obInes.ert it unil it won't go any 9.1.8 aGpportooprtihaete"Emdaistsoer"s p1a0g4e2,7anadndse4t9I9o,nization Mode to ES', and the 99..11..910ISfittrisAncoqtuiisnitsiionng.leAsiosnigmnoadefilge0natome",OpMtiOo-nD"AanYd-sYeRt S+IRl.etter. Record it in 9.1. 11tRhuenlotghebboeoek.f liver samples first, running each standard twice at the 5be1g0i7nnsianmgploefst.he run... Run a QC check by running one standard after every 9.2 9M.a2n.u1alDrIanwje1c5t0iounL of sample into a syringe. Inject the sample into the rheodyne 9.2.2 Tinujemcttihoen vpaorltv.e Itnoje"cOtns"l.owly. Record the sample ID in the log book. 99..22..43 WReaciotrtdwtohemisncuatnesn,uamnbderinfjeocrtetahcehnseaxmtpslaempilnet.he logbook. 006346 3 9:3 9U.s3.i1ng StehteupAustaomsplaemptlraeyrA, B, or C. 9-3-2 vRieaclosrmdatyhebesaimnpelaecsharnund, thir positions in the instrument log book. Up to 17 9.3.3 9S.e3t.u3p.1the sampler: Push the sample bution 99..33..33..32 SSeetrsinjaecmtlsoapomppslliez=ee2= 100uL 99..33..33.45 NSaemteCytchelfeidlem:eLi=v0ers 99..33..33..76 9.3.3.8 AIddednttihfey sthaemptlraeysutsoeQdueue by pressing "Enter" Press "Run"tostart 10.1 1Q0u.a1l.itRypueaCnkonahtesritgoahlntdoacrcduervsesrtyop5t1h0er7usna.mpOlnelsy. tIhfea ssaimgpnilfeiscabnetfcorheatnhgeel(ast50ac%c)epitnable standard wil be used. The remaining samples will be reanalyzed. 10.2 1Pr0e.c2i.s1iSoeneaMnedthoAdccVualriadcatyion Report numberAMDT-M-5.0.1 10.3 10.4 ReOftehretroVMaeltihdaotdiVoanliPdaatriaomneRteeprosrt Number AMDT-M-5.0.V1 1111..21 CPlaoltcutlhaetisotnasndard standard, curve, using the mean of the two values obtained for each 11.2. 1rReegraedsspieoankthoediegthetsrmoirneartehaes sfaormptlhee scoanmcpelnetsraftiroonms.the printout, Use linear 11.2.2 Clailvecru:late the mg of FC-95 anion, orotherfluorochemical in the total rabbit `mg FC:95 anion in the total rabbit liver = megmFsC-of95liavneriounsfedrofomrsatndalcysuirs,ve x Total mass of liver, gms 1111..43 sPMrtaiankntdearacdrheIrsDou.lmtWasrtitoatgberleathmaenfsdotreucndtayecrnhuistmaibmneptlrheeo,nswtituhtdehyptbrhoienotkpo.euatk,sinliatbieal,eddatwei,thantdhepsuatmiplienotrhe study folder. Staple all chromatograms together and number pages. 00347 . None 13.1 AMDT-EP-17 LOREVISIONS ~~~ Revision Number Reasonfor change. 000 Revision Dae 00348 5 3M Environmental Laboratory Method Analysis of Fluoride Using the Skalar Segmented Flow Analyzer With Ion Selective Electrode Method Identification Number: AMDT-M-8 Adoption Date: /o-s-g Revision Number: 0 _-- Revision Date: None Author: Deb Wri/g Cynhtht ia Weber Approved By: up Leader 4 4 ps Zid Quality Assurance /o/ls/2s Date 9-27-25" Date Software: IBM MS Word, 6.0 Affected Documents: AMDT-EP-26, Operation and Maintenance of the Skalar Segmented Flow Analyzer 00349 1 BEST COPY AVAILABLE L0 esc1o.1re0 This method 0 is for the analys0 is for fluoride0 , thermally ext0 racted from sa0 mples using the Dohrmann DX2000 (AMDT-M:-1), and collected in TISAB for analysis with are Ion Selective Electrode (ISE). The analysis is performed using the Skalar Segmented Flow Analyzer with ISE. 1.2 SthaempDloehrsmcaannn.be tissues, serum, biological material,or other materials extracted on 2.1 Skalar, segmented flow, fluoride. 3.1 Follow standard laboratory safety practices. 4.1 Supplies 44..11..12 ASuatmopplipeectusp,so,x4fmorLdpolraseqtuiicvcaluepnstwwiitthhcpalpasstic tips 4.1.3 4.1.4 Polypropylene volumetric flasks, 100 mL Cartridge components, refer to the Skalar Methods for components and part 4.1.5 Snaummbpelres.prefiliers, Evergreen 4.2 Reagents 4.2.1 Brij 35, 30% S.F.A.S. Detergent 4.2.2 TISAB II buffer solution: Purchase TISAB II from Orion. To 1 liter of TISAB II add 2.5 mL or 100 ppm fluoride solution and 1 mLBrij. 44..22..43 NSiarmrpilcearcirdinssoilnugtisoonluftoirond:ecDoinltuatmeiTnIatSiAoBn,II1 1N:1 (wliatbhgrMaidlel)i:-QSlwaotwelry. add 64 t`hmeLsvoclonucmeenturpatteod1nLitrwiictahciMdil(lHiN-QO,w)atetro.250 mLs of Milli-Q water.Bring 4.3 4S.t3a.n1daSrtdosck solution, 100 ppm F: purchased from Orion. 4.3.2 Intermediate standard, 10 ppm: Dilute 10 mLs of stock solution to 100 mLs with Milli-Q water. Use polypropylene volumetric flasks. 4.3.3 Working standard: Make up the Following working standards by adding the volumes of intermediate or stock standard indicated on the table,using [Worki0n00g0pSptammndarwidth| ME1 O wamtLers. ofStockSta5 ndard |mlsofTn3 e5rm1ed5iateSandard| oxford or pumpmate pipets, to 50 mLs of TISAB and diluting to 100 mLs --oo 0p pm m-- 5] 15 ppm 103 00350 a C-- Tsppm70s ------ | 5.1 Skalar Segmented Flow Auto Analyzer SansTM System equipped with ISE 6.1 Hinitgehrfceornencceenst.rations of alkalinity, chloride, phosphate, sulfate or iron can cause 7.1 wSiatmhpilne3s0shdoauysl.d be stored in polyethylene bottles. Samples should be analyzed 8.1 8P.r1e.p1araPrteipoanreocfalCiablriatbiroantsitoanndSatrdasndasaridnssection 4.3. 8.2 8C.a2l.ib1raTthieonstandards are analyzed at the beginning of the run. 8.3 8S.t3o.r1ageStaCnodnadridtsiaornesstofroerd iSntcaanpdpaerddpsolypropylene volumetric flasks. New standards are prepared at a minimum of every six months,oras necessary. 9.1 9St.a1r.t1 UCplamPrpodcoewdnurtehe pumpdecks, air bars and sampler-pump tubing. 99..11..32 TPuutrnthoenfltuhoerpidoeweelrecotfrothdeessainmptlheere,lpecutmrposd,e cohffasmebtepro.tentiometer and heating 9.1.4 Pbautth.the reagent-lines in the appropriate botles. 9.1.5 tTuurmn oonnthteheintienrtfearcfea,cceombpeuftoerre, dtihseplcaoymapnudteprri.nter. Make sure you 9.1.6 Let the system stabilize for approximately 30 minutes. 9.2 9St.a2r.t1inCgreaatReuansample table by selecting FILES, TABLE, and CREATE, type in 9.2.2 Ptrhientnathmeesoafmtphleeftilaeb,lea,nidnsperretsesdEiNnTtEheR.system table by pushing ESC, 9.2.3 DPiRaIlNtTh,e GsaRmpOlUerPse1.ttTihngisstwoiltlhepraipnptrotphreieantteirneurmubn,er of samples, number of 9.2.4 Fsielclotnhdessfaomrpslaemtprlaey wwiatshh,thaendstnanudmabredsr,osfamspelceosn,dswafsorhetsheasnadmpdlrief.is. IW and 9.2.5 SFeWt /thReUbNasOelUinTe.cups on the sampler do not need to be filled. 00351 3 9.2.5. mISaeylebcet iGnRtAhePHDaItCaSH,anRdElAinLg TPaInMeEl.. SIwfiytocuh cwaontnhoetAgneatlyrseials-tPiamnee,lyboyu 9.2.5.bs2eaUlsseecetliitnnhege. sCAmdOajlNulsTtsRctrOheeLwbdaPrsiAevlNeirnEefLournattnihldeipotfuifssshaeitpnppgrooFtx7e.inmtaitoemleyte3r/4toisnecthtfhreom 9.2.5.3thCehbeoctktotmheohfitghheesstcrseteann.dard the interface screw #3. and adjust the gain, if necessary, with 9.2.6 Gwioltl onoCtObeNTruRn.O(LSePleAcNtEoLr,deasnedletcotaannaallyyssiisspbayneplr.eDsseisnegleEcNtTtEheR.an)alPyrseissstThaatb 9.2.7 PtroersesttuhmetsopathceebAanraltyosbirsinPganuepl.the local menu. 99..22..98 TSeylpeectySouTrAIRDT(itnoitsitaalrst)t,hteheansalaymspisl.e table which you created under 9.2.1 (or aprnedssseEleNctTSETRAfRorTcAhoNiAceLsY),SIchSo.ose running with or without the system table 9.2.10sAefttetortshtearrtiignhgttnhuemsboeftrwoafres,amstpalretsthaensdatmhpaltert.heMsaakmeplseu/rweatshhaatitrhetismaemsplareer is 9.2. 11SOeKl.ect GRAPHICS, REAL TIME to view the progress of the analysis. 9.3 L5o3a1di1ngGotaondCOPNrTinRtOiLng, PthAeNEDLa:tar-Feislehe spacebar 0 bring up the local menu ahingdhlsieglhectttLheOAfiDle.toSebleecptriAnUteTdOaCndApLrCesUsLEANTTEIRO)N.and enter the filename (or 9.3.2 TCoURvViEew. the calibration curve, go to GRAPHICS, CALIBRATION 99..33..43 TToo pprriinntt tthhee lhoigwhlleevveellsccurreevne,, ppuusshh EPSRCINtTo gSeCtRoEutEoNf.graphics. Select CSEUTRTVIENGaSn.dCphreasnsgeEStChe, maanxd Eyntvearl.uePrteosaspPprRoIxNimTatSeClRyE9E00N..Go to CAL 9.3.5 pRreetsusrnENtoTSEERTTanIdNGPSRIaNnTd cShCaRngEeEtNhetmo aprxinvtaslauempblaeckpetaoks4.095, go to EDIT, 9.3.6 TOoUTpPriUnTt.theSerleesculttsPRgoINtoTCERONfoTrRthOeLEpPsAoNnEoLr,PSRPNACfoErBtAheRL,asOerU.TPUT, 9.4 Shutdown 99..44..12 LPuettatlhletshyesrteeamgernitn-sleinfeosr ianppMrilolxii-mQatwealtyer3.0 minutes. 9.4.3 oAffftseerttphoetesnytsitoememthera,s rTiunrsnedocffomtphleehteealtyi,ntgubmatohfofnthweeseakmepnldesr., Lepauvmpe alniqduid in 9.4.4 Tthaekelintehs.e electrode out and soak in 100 ppm F overnight, 99..44..65 SReelleecatseFItLhEeSp,umprpe-sdsecAkLsT, aFirabnadrssealnecdtsQaUmpIlTertopeuxmipt-tthuebinpgr.ogram. 9.4.7 On Friday, ur off the computer, display and interface for he weekend. 10.0 VALIDATION 10.1 Q1u0a.l1.it{yRuCnoantsrtoalndard (mid to high concentration) every 10 samples,If a siacgcneipftiacbalnet cshtaanngdeardinwiplelabkeheusiegdh.tTohcceurrse,maonilnyintghesasmapmlpelseswiblelfboreertehaenallayszted. 00352 4 10.2 P1r0e.c2i.s1iSoeneaMnedthoAdccVualriadcatyion Report number AMDT-M-8.0.V1 1100..43 ROetfheerrtoVMaeltihdaotdiVoanliPdaatriaomneRteeprosrt Number AMDT-M-8.0.V1 11.1 CI1a.lc1u.l1aTthieonstsandard curve is plotted by the Skalar sofcware. 11.1.2Abeltltecra.lculations are done by the Skalar software. * should be 0.995 or 1111..32 WPrreiptaertehespsrteuaddyshneuemtbsetro sounmtmhaerpirzinetoduattsa,. Iinnictilauld,edastaempthleepvroinltuomuet,, awneidgbhitnsdutsoegdetehtce.r swiutmhmaallrypaschkeaegteanddocpumeenitnstoatnhde psltaucdeyinnottheebosotku.dBfaoclkderu.p Malalkdeataa caonpdysopfretahed.sheets 11.4 EolnetcotrsotnuidcyDdaitsak and backup disks. 11.4. 1sGtuLdPy,satunddieasl:soEldeacttaroinsiccopdaiteadiosnctoopaiefdloopnptyodtihsekStthuadtyisfsltooprpeyddiinskthfeorlacba,ch 11.4.20ther studies: All data is copied onto a floppy disk thati stored in the lab. 1A 2No.n0e TTACHM ~~~ E ~ N ~T ~S 13.1 'ADMXD2T0-00M.O1rg,anTihcerHmaallidEexiArnaacltyizoenr-oLfiFvleuroride by Means ofaModified Dohrmann 1133..32 ASkMaDlTa-rEMPet-h2o6d,s,Op#e3r3a5t,ioSnkaalnadr MMaeitnhtoednsanMcaenoufalthe Skalar Segmented Flow Analyzer 4.0 REVISIONS Revision Number Reafosrchoannge Revision Dae 00353 5 9.3 Quality Assurance Unit Statement 00354 Atiachment D GLP Study Quality Assurance Statement Study Tite: SingTl-e6-0d5o1seaDnedrmTa-6l05A4bsoirnptRiaobnb/iTtosxicity Study of Study Number: AMDT-013195.1 Name of Auditor: Kari Rambo ``TThhiesfsitnuddiynghsaswebreeenreipnosrpteecdtetdo tbhyetshteuQduyadliirteyctAosrsaunrdanmcaenaUngietmeansti.ndicated in the following table. Inspection Dates Date Inspection Reported to 1001205 101995 Final Report 101995 10/1995 oe Cr _p0-15-55 AU Auditor Date 00355 9.4 Key Personnel Involved in the Study 00356 3M Environmental Laboratory Key Personnel 3 Thermal extraction followed by analysis using Orion ion analyzer: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Analysisofliver extracts using electrospray mass spectrometry: Jim Johnson Dave Christenson `Thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Documentation and Reporting: Jim Johnson Rich Youngblom Quality Assurance Unit: GCyanltehViaanWeBubsekrirk Kari Rambo 00357 9.11 Data 00358 d9.e1t1e.r1miSnuemdmbayrtyhearnmdalraewxtdraatcat;ionugfoFl"lionwewdhoblyealniavleyrsiass using Orion ion analyzer. 00359 Summary of Combustion Data - Liver AMDT-013195.1, EWI 6329-133 As Referenced in Final Report section 6.0 DATA ANALYSIS `Total ug Fluoride in Whole Liver Mean per Dose Group* Bg Std.Dev. Control Group 162 + 5.5 0.128 mg/kg dose (T6054) 11.1 + 2.0 1.28 mg/kg dose (T6054) 17.1 + 2.6 12.8 mg/kg dose (T6054) 45.2 + 205 Fabric Exposure (T6051) 14.7 + 2.7 "Calculated as the mean of triplicate samples from each of three male and three female rabbits. 00360 FC129 AB n pAcotmuFa-l % inliver liver blank-3 fey (0W23W0) ilvveerrssppiikkee-ss 8833%% 019.5060 ivveerrssppiikkee.s7 19042%% 11.0208 Iivveerrssppiikkee.dd 8834%% 224394 Fli5ve2r8b7i3a1nk.4 00234258 (FFs52288773332 0033230% FFss22888855.12 0042429 FF5s22680885-.13 00.310369 FFss22889988..32 00.11033 Tlivveerr lbalnakn.k12 00229288 iievresrpsipkkeet2 9951%% 01.93965 FFs522887788-.12 0022808 FFss22887883.13 00211716 FF5522088833..32 0012729 FFs522006677-.12 0011548 FFss22808677..13 00220502 iFvse2r88bl7a.n2k-1 00218819 Iivveerrssppiikkee-t2 6859%% 00.996783 ivveerrssppiikkee-d3 18039%% 01.28795 ivveerrssppiikkee-s5 110011%% 11.2161 FFs522888837..13 00..116783 FFss2288093332 00117457 ApveprmaFg-e i(nWfiWv)er 0303 038 0124 021 0211 018 0205 0163 liver Wlhiovelre (bgurmamesd) (wgeaimghst) 00.113420 00.12118 00..114028 0010118 00112017 774422 00110504 670422 0o1i0e8 2602 001s50 773333 0o.u15s2 733 00..110314 00.1104s5 702 0011s5t0 770022 o0i1017s T73E8 00113445 671358 00110186 e6i1ss 001108 770088 00.112098 00.113423 00.113288 00112418 708 00113353 66335 5 013s eas TotwahlolFe-in I(uvge)r 224 28 ea 1e2 158 "a 1s 104 RPTIILXS D(omsgakgge) 00 00 00 00 00 00 oa 0128 Page! G0G361 Fos as FFarieassneoorrs2t FeFFeeoaaooorrraeeszt FeFFeseassmseaazs2 FFeFizezszo0sororr12s vCveeerrrbohit1k2 eLveerrroosikt2 FULavvzoeorr0ooeptpkzs FFFoeizzssoeoosrsts2 FaFFseesozooortnsz FeFjseesaoeseatrs FFFaerzasonaeoeses1z FaFFssezzoaoa0on0e1zs FFjasareseod PFFoeazaassneoos FFeFeseszssseeensrz FFeeamsemoes % pAocmtEa. nie ApvearmagEe iver buvmeerd Wvheolre ween oy (o0W1e5W6) (WW) ome (goTriassmss) (g7Ta3am4es) o01t7s9 Gis ome oazrss ome 7aa4z az aoheiirssd ous aorses oi 7aaz: 781 0wG1ii2ar0 ot oamlo ous 170018 708 00o23r1se oacttieiar 108 o2i 03280 aoGrriassn oaswwo 0 O1s1a0d o0o11r2s87 00223s8 az oats ooites oer eeaarr oer 0o02i2%@% ozs o0o1i2ts a6a4isss o002z37o%3 oz oaotiroser ea8a8ry7 ao02zz08 oa 00o11i0se1s 6a85s50o0 Go023m02 oz Ocoiimsnee cceeao es M050 02% om ooiesr ots eeda ove 0ac5es7res ose cooirisye 6ss 506 0oc4e2rn0 oso oooiensrs 7775ss3 o0G3li4ea0 owe ooGim1es0 1sot a1 Totna aFin wer (ug) na em mo ez 1s0 us mi 1s0 ms mi ma aso mr RPTISSLXLS Dosage (mark) ome ous oss ows 12 1a 12s 12s 12s 1a ne 12s ne 0036pogo %2 Fci29 a8 PAcOtMuEal. ApvpermaFg-e liver Wlhiovelre fiver blank 1 % roy i(nWlWve)r 0265 i(nWliWv)er (bgurmamesd) 0125 (wgeriagmhst) lliivveerr bbllaannkk 23 00232207 00.111213 ilvveerrssppiikkee--12 08.777%86 01..90251 Iverspike3 0.7852 0.823 00.111423 0.144 llivveerrssppikkee--45 00..8946856 01..80569 00..114398 Fi5v2e8r8s0p.i1kes Fs20802 82% 00897182 06 110 00.111247 0113 e6e877 FF5s22880849-.13 018.3505 00112417 764807 FF5522880944..23 Fs2805.1 00971974 oss o0i11189 774400 0307 0151 581 FFs522080955..23 00257283 038 o011s0e0 5s8a1i FFss22887744.12 00323419 0251 0011424 669977 FF5s22887704..13 00.117638 00113445 869917 FF5s2288779932 F52063-1 00.113845 012 00114075 889911 0216 0130 702 FF552200663.332 00.117554 ot 00113253 770022 FFss22887777..12 F52877.3 00128202 ois 00111189 e6a0sg 0.180 0141 699 FFsa22088e8e-2 Fs28883 00.211518 0170 0011044s 664422 0142 0139 42 FFs522096666-.12 00322117 0285 00113213 667788 LFi5v2e0r8sp6i3ke7 60% 00.68871 00.11:022 676 LUivveerrssppikkeesd 7715%% 01..70755 00..114083 ULivveerrssppiikkee-1110 18083%% 01..91950 00.113580 LUivveerrssppiikkee-1132 17081%% 21.1166 00111303 LLiivveerr ssppikkee--1t54 110027%% 22.3495 00.113310 TowtahlolFe-in Iver (ug) RPTISILXLS (Dmoasrakg)e 754 128 628 128 214 128 175 Fabric 163 Fabric 128 Fabric 183 Fabric 108 Fabric 173 Fabric Page 3 00363 9.11.2 Summary and raw data; analysis of liver extracts using electrospray mass spectrometry. 00364 sean iza JsPTrureetsumber: T-- bheese a-- ust r-- as 0120) fRRr=aaaetrsFaausonoAmn, SanaieeeE FCHrarEanT fofprdirs-- AC)ohntAalaogmn o-31-a5 O. Arlengan ; So wCao e BT[Thaeeaw R= an Me Coamaane | rBmms wo RaaEe Nowe O er RgC er AeS e STFoosoHpuer NIIwe] w4vweo JIo w" wNIwoe] wNIweo B BRwaamewamdwWeee idlteameeeem 1 11 Goaowewmese aoEwmmmw oooomxow fe B Baaedws m lE ow 1 E1 oGme gGmm oome RBhaaeee mmeeww memwsw 111 oooememe ooammm oo=om BBwEEaEaSnT mwoGmemw oumwe wm 11 1 wooimmse mamms ne ooo1om)nn mw os *eIlhrcoornnecnhnsracilc nAhTe co 5o.bytToiaNCSs ncoaPhnanRnicoualncnurb4vei abdnsdamruslcloAidneyg Aea ssceebyS 4o258,rTaoeA4t5n ved 600365 HWI #6329-133 fPsrTwreaasoynt:toNuamber RRResoasiovrsses FVaeuer:Amount DeBuinto:: Uisese rue FTlSaoooolsnG7oes08Do1rmaa RAsartpsio(nC129 9oo3e0i508 pCiFoesrrsnVse 200 hctoprMeyS I aa a a foemman | ee r vweA wes rTaomuers | Fmmem mwee treene eae we 1 ovmoe Tree 11 oeoewe | aes=s = Pome wn er 1 oo we ae F-- osse 0V00o0 ooosn aos * Administra328 10 cm x 10cm loco of tat arc TiSThahsiiccrdaolynocc.uhalraBrteYuonsEiotlnhIwra4sNhcIaSnlttc7hAu1AC1eR3IHbdOyTnahueIsiCncngoatnsnchacPsntaaarlnldolanerdsnicuh5rhv4nsanhnrdomdsudlcclAoionnWgosytemhBcroeeso5t3rbtys45o,e Tmhe4od5 t,o In 00366 Fit cuodasa cepior Metiod DLELIV Az Run date CSoaumtpaletorDLCorLcIV 05-09-1995 05:43:44 BEST COPY AVAILABLE Version: 12 StraiPgrhitnteLdinsonFi0t5-0f9o-r1c3e9d5 tAhTrou0g8h:49Or:i5gin. 7 639-143 Rymere 6329-133 A RITE i A ; Component $11 re yd rd Sid yd / mT yard ps / rd 4 3 12 HINT EVEL AmounT EXTERNCAoLmpoSnTeAnNtDARD1 CA=LIBRATION AREA :1 5 03.142000000 1.2000 5457673433 132053 . SWOPE + x + InTeceeT AmouAnrtea + R squarsd 5+ .101i.20E3-8086E.0x5 = 0.9916 AArmeoaunt ++ 00..0000000E5.2000 00367 we rFiEe: odomsr0as Reg= Eeple+ estonmaeseD,LoLve.rLes BESTCOPY AVAILABLEAs-e Run Puraitnete0d5-o0n9-10959-509-0179:9254:4A5T U0e7r:z2i4o:n5:4 Lo Straight Line Fit forced through Orfuin -. i? 3 i i | i pi 8 | 49 || Component 1:.1 + / 3& g D> TS 1 f . | 4 8 12 AAT CEvEL amount EXTEARRNE-- AAL ST-- ANDARD CALIBRATION ::i 2.3171342000000000 1S3523310049075 v= SLOPE x + INTERCEPT AmouAnrt.ea = R Saared 0=11.85638.78765128075E-0+a |+ fArmeasu.ns ++ 00..0000000c0~8000 . 00368 Ea WaT (329-133 iz< Reg _ ECoc pyofOt riginal. -- asfis x /FSeimpllele6%38428939-51D7" 3 RABBIT LILVAEBR-BEAXSTERA~CTTSh:e MFSo-Dia3t9a SyCsastsem 00 Le nee oT bers o WALA SAAR Relld or fe, 83/84/1995 14:00 ae po =e & . 28 13 Jig SOm22LeE, osT qm ee ae omen | we = mr | + 1% Beef Iver actods L wl. cue, SFaimlpol:eo6a2a2zs5-133 RABBIT LILVEOR XBTA-~CTEBShSeAHS'SSDCaoEAt3aS Sy"tsatnesm 100. TOP ses rs. ' To Hg L 18 3 : 4 , | 03/04/1995 14:08 2g) ys" 4 g Last TT | Lo ge ane arg HE sc? Beeen iy a= | | i)iv A. SFaimlpeo:a16z3s25s"3 manny 1L1B0SSSRETIRCETShSo 1"S BDeatsa Syasetson oe sr Em bers: oPLE : bn B 3001995 10:00 : . - a Lgl Eo heee Es Tm 8 lors 3300 & we wa ww wy | isYU Ca SaFmilpol:eo46a332s95-133 RAIT LILVAEDR-BEASXETR-ROTThSe HPSoD-a1t5a Sutsats)on 100 aes Ts oe wr 612 fd A gL sol Ef] oe EL ELE ELE] 83/04/1995 14:08 ws f l 5 ars of ase} E 35g OF 43 00oe ww xa.9 ~ SH .i brs. was | 1 . I asx ave fe ,. FSialmo1:0t6o397s5n13 namn1 LiL0AEBnDABSXETAR~CTTShSe M"SFDoaetsa Su"rtanosn 03/04/1995 14:00 ee 2 apr fe re 4 hi M sie S35 ses Som - 71 wel]? sho) EEN ECER loElHEEl OE O|E 2 100 263816 wos x6.9 ~ Si [ bers. 8g we wm lso8 = = ws 7 = 78 199377 ve Su iQ- hot63(202 s13e3edAolmlsay AndeapbpeeD o=E:.ciriC aeloofp cDOprearigy oreinmsal ~. faoenmr SFeimlpol:eot6o3s7o9s-Hn13E3 (FG-129)L8AD6B3A7S9E-14-3Th(oFC11S30b3aFt)a MSAyRsDtIoTn LIVER aAaT/HmRaC/HtSons 10:67 100 agg 467 ou hrs sogg50 I2)e 611S,N 4 ga of a 24 ET SH : 32 9g: 100) 396.0,re 30%;2 lsd re og = = sues Hh <3 ,. FSiamtmioe:ou0n3t2o5s"s130 (ro-1793'L 4 cashB e--sThLeECMD SSkDIaRt)a SSAyBsStToEn LIVER BbAamPaAsiCoss 10:07 roe boa" bm MHI agg bs *EgePT waame | aarm&e HEE 4 WR, 3996 n 222 fom ne sea eaig is d9.e1t1e.r3miSnuemdmbayrtyhearnmdalraewxtdraatcat;ioun gfoFlilonwwedhoblyealniavleyrsiass 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 theconclusion stated in section 6.0 and therefore is not discussed in detail. 00376 RE: 6329-133 LIVER SAMPLES AMDT 13195.1 Date of Analysis: 3/28, 3/29 and 3/38/95 Analyst: DOW Uw lklas SeonQata The samples are burned in the Dohrman at 958 C using between 8.1 and 8.2 grams of the liver. The gas is collected in 1.8 mL of 1:1 TISAB/Milli-Q water then an additional 2 mL of 1:1 TISAB/Milli-Q is added to allow for sufficient volume for Skalar 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 and 8.15 - 1.58 ppm fluoride. The standard curve for the high range was plotted using the inverse logarithm option. The 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. 00377 by \ J, 29 SUMMARYor 6320133 LIVER SAMPLES AMDT013195.1 STE o AILABLE Thi rl Alls se oeem ts. DOV wTFmwEo wBOwN wTdeeI BoRw we Fm wigwR w . ggi TosaoowwenNoaoewmwBw wdaEowuwoR 3 i anouns mTia oWmm Bow ouwmm wowo om A ame ww wToin moBw ONN0 ooaeEmoewOR we [a4mi2]w mowom n ommeomwe we owuAs ms wTmheaerlowwBwBoN ooowmme oonwBo swasmno.owRw ' 13LSUM } Page1 summrNsEnRysSrueaesso133 BESTCOPY AVAILABLE aEWewR0 mwmomow ow sawmnow :g waaoea BWEOWNONoR MWRE om TowEIwoMmBb EE &2: BmAEOow OonN uummolomeoe owoaummdor. HBRSolOoWORR mammoo oLeoomon3 pte se RwASwNMoEm wWCU w Baame 2ww mmeErOBwmON oomunom ovow oowmemonww - f-- et Liss 23Ptwebea o dm oi0nowmw e s sSasat oonms oame owmw oe om Ts5 sseafae woomnm oom neam oon we BnBooooS Ssaa oodnee ooimam dooommm o BW 1 sowaave 1% nLwoe aoww gSEukn ammaa wwo 0 2 annv seoaho owoo unBoo rermemmesar w 5B7 ohwwaes ue iwwow we N5Bo rarraasmwa 0wo ARamamss rasas wwo wo N 3% a Erweess 00 wo ~ 3> :33 S5Sooadomsm owwoow aaammmsss ooowww S5ooowss WWww daaann woow 5sooome WWwoow awmees w 335ooooa emns WWwoo waawmms 3JSooomm Ww awmiee Soom wo mae Paget BESTCOPY AVAILABLE wwwrnn & ooonns ow on m mwm he3!: Ssa; 33: a= sins n3 omoom om m w w > <g &&& BESTCOPY AVAILABLE : Page 2 umans FFLII ome e Amo o4 LoCI Oo sm ewmsodwomm om SESTCOPY AVAILABLE bFSO ooodSa no oaAme NR o0oooamn om BBWooootooessn R ndok mimn m gPgnEn oomm oonon 2ff] g8nBFnI oBwwikm Bh o0oon0ni2 33320 Boa ow aw ow ooGmme SBommm waasonooasns dweormn ENF oa s ms ome oJi 18 ow BBNoootooenm owMhoo ddawmseua1hb vFF A os m ooonn FC IH noomms oonw aasEs mwmm FH 5 i54F Fat 5i23 - Fi Lssaxs 1Vebew 5FR ssea 6Tsossasao WW5o SsSas SSBuBossteawn 2SG5ni oowmmaaa SuBogoama 2noHpSaaa 3nFoR mSaa B E BT Y e o bo rraas n3no arrwamea B N5o . arrawemsa ~ 0aw 0ma owew ooonmms ooowmes omomww ooomnn oooosmn owowwm ooiwam oodam uweoaw xio wndow omww w0wn 350 3 000000ss 230 02000%66 bob3dy ae apaormt owe 3 ws ow ooooon 302000 ooomwmms oo0xn% oN00Dos 3S0o0 0aa0mw0s AiWnuD aaasn0 SWaomaohms iainnn peas een an mgwieennme uusomwn awaoemes amNnDa Gnmeoemwme wemeew BEST COPYAVAILABLE owwknm aGalmm aeaskk ooonnw um om wwwnoo GoGmmm o0osss ooamnn abdmee LEEYT: 8vP0eS3t iT$a5g1 2 sans P:DPooEa8mm Oe 8m zBomx0 aumn oesem soeaw wn pIoEm . 8. -3322 w: Page2 1995-03-28 17:3*0 outkut of : 95032681 Software : version 6.1 190,33 operator + oow Date of the Analysis : 1995-03-28 15:28 Analysis File Name : Ci\SKALAR\DATA\350326F1 200 hss JHoaTp 0T3A2iR9A4S 13 3 FClauloirbirdaetio1n.5order = Inverse Logarithm s1ope sa An Result - 10O80 x%sia-sci11 aiz.= 0T.o0l0o0n0o0 Wo lel xG51 === ScClooorrprreeeccottfeeddtvvhaaelluueeeleoocfftrottdhheee csoanmpcleentration i FClauloirbirdaetioLn order = 2 Correlation : x = 0.99716 Result = a2 + x +al * x +a0 aazI= 0o.l0o0o0o0a0z . 201 olaosos sampler TyNipmeber Simic Tine +iis5a0t00s0ec. VMTaaskseh upTTiimmee 1: 120 sseecc.. Single SfpeeecdilaelHeight | N70o.nemm. Diluter nGSeiieTldaulcreiiooHnneivFgoahlctutmoer :: : 8100 2.5 mm ml. RDeislaintpiioen runs i1E1 URseeprrocfeisle : No . mr 00385 1995-03-28 17:30 output of : 950328F1 Fluoride 1.5 Psaitghnalnumtbyepre :: De3 bubbled DescyosltoerNumber :: Y0es dRieLsuatmeple :: NNoo dQiilG Touhtrpeusthold :: 40095 Window event : OFf ss12 ssTTaannddaarrdd :: IIggnnoorree 5s3d ssTTaannddaarrdd :: IIggnnoorree 5556 ssTTaannddaarrdd :: Igno0r.e150 5s7e ssTTaannddaarrdd :: 00..360000 85910 ssTTaannddaradrd:: 11..250000 ODridmeerns:ionInv: erPsPeM Logarithm . sttrairgtgevralLuiemit :: 5108000 DSeUc Psteaarktshiagpneore : P60ointSeedc Meenadsureigwnoirnedow :: 71205%Sec FRielgteenreration :: NNoo ofourntuplaut:: HE. HE Fluoride L SPiagtnhalnumtbyepre :: De0 bubbled DesycsotleomrNumber :: N0o dRiersuatmeple `: NNoo ddiilG Tohuretsphoultd:: 04095 Window event : Off 00386 1995-03-28 17:30 output of : 950320F1 5s21 sTsTaannddaarrdd :: 00..001350 5543 ssTTaannddaarrdd :: 00..009600 5s65 ssTTaannddaarrdd :: 00..115200 5587 ssTTaannddaarrdd :: IIggnnoorree 55910 ssTTaannddaarrdd :: IIggnnoorree Doirmdeenrsi: on2 : PPM sCtrairgtgerValLuiemit :: 5108000 DSUec . spteAarktshiagpneore :: P60ointeSdec MeeNadsureigwniornedow :: 71205%Sec RFeigletneerration :: NNoo ofourtmuplaut:: cH4:E=cEd 00C387 1995-03-28 17:30 output of : 950328%1 FFlluuoorirdiedLe1.5 Page 1 of 3 Ppeoxt Pos Typ Ident Dil Weight Ch Result F Cor. Valu Time : wt iw Initial Wash 1 1.000 33 00..0005660 0o 215To 6sTo 1c Tracer 1 1.000 330.15.715969 22006655 22820 203 TV: MTpp 2a Drift 1 1.000 340.19.928310 2200996 23120 380 TV: Llpre 0 3 ow Wash 1 1.000 43 0.00.005660 0o 214To 557To 4 si standar1d 1 1.000 330.00.105682 4433 2580 7320 5 s2 standar2d 1 1.000 330.00.300730 110066 3220 907 6 s3 Standar3d 1 1.000 330.00.507858 221144 4320 10810 7 sa Standar4d 1 1.000 43 0.00.819039 332255 S5404 12570 8 s5 standar5d 1 1.000 3100..21355 444a00 6610 1432 5 s6 standar6d 1 1.000 33 0.01.416552 S50066 7280 16060 10 87 Standar7d 1 1.000 320.02.927953 889955 11210 17810 11 s8 standar8d 1 1.000 330.05.65084 114a1100 16410 1956 12 s9 standard 1 1.000 330.15.527289 22009055 23320 21320 13 s10 Standard 10 1 1.000 33 117.54773 22333358 25760 23050 1a Drift 1 1.000 33 1.10.020488 2211133 23380 24820 TLTppe 15 ow Wash 1 1.000 43 0.00.005660 o0 226T2o66o1 600388 1095-03-28 17:30 output of + ss032erL age 2 at 3 FPlluioorridied1e1.5 saon Pos Typ dent 6u BK1 Dil 1 Weight 1.000 Ch Result 3Toltbeerote< Cor. 114 Va2l4u02T7i7m9eSN MTT om' ou Bika 1 1.000 3 AbsenA -32 134 3007 . 4 oRHEE -32 0 o Lg ou Sx1 1 1.000 3 AbseAn -23 202 3102 4 0.0011 -23 0 o ~b ou sk2 1 1.000 3 AbseAn -33 192 2387 20 u 4 ade -33 0 0 Soka 1 1.000 D3 o0l.0s87 d20e9 436Y03U5T3D3 b ows . nou Ps250-1 1 1.000 3 tools 17 242 376 4 0.0098 17 Bd 2 ou ps2000-2 1 1.000 3 tooe< -1 224 3820 4 0.0058 "1 0 0 vb 2 uw rs2000-3 1 1.000 3 Absen A -33 132 4057 . 4 #.HEEE -33 0 0 # uu psase-1 1 1.000 3 0.08 12 236 416 4 0.0087 12 0 Do su Fsaseaz 1 1.000 3 AbseAn 32 192 4407 . 2% a Drift 1 4 hoHAEE 1.000 D3 a1d.2e59 -32 0 0 n21a24 2348UN 4s81 Ternppe 2 ow wash 1 1.000 3Po0l.o05s6 00 226047%51 . 2 ou Fs2066-3 1 1.000 T3 otohoe< 33B2 256 4871 w 29 u F52884-1 1 1.000 3To0.0i61 333 256WT50R89 "0 oxw 2 u sasee-z 1 1.000 3 0.06 34 286 sama oF ot nou 4 0.0135 es20se3 1 1.000 3 0.081 34 o 4 256 5433 AB us wsA 4 0.0135 34 0 0 / 00389 1995-03-26 17:30 PososTyp Ildeenntt Dil 32 wu Psas92-2 1 Hu ps2ses2 1 34 ou Fs2eee-1 1 FE ps2963-2 1 36 u Fs29633 1 7 uw esgesace a Bland TIA 3a rift 1 3 ow Wash 1 we rw Runout Wash 1 output of : 95032081 FFlluuoorirdiedIe1.5 oEorn Weight Ch Resuullt FCoorr.. 1.000 P3 o0l.0a61 3388 1.000 3 0.061 39 4 0.0147 39 1.000 3 0.061 36 4 0.0140 36 1.000 i3 o0%.n0601 3de6 1.000 I3 o0l.0a61 336 1.000 3 beena 23 4 0.0011 -23 1.000 3ak1i.2n57 5212232 1.000 31000.60506 00 1.000 i3 000.0056 00 Valu Time 260o5o635 260 5805 0 o 256 5583 o 256o6e159 25606%333 155 6506 0 2340TNT 6684 2180 68580 0o71o59 Page 3 of 3 32 g. mHpA oa%: "m = ee a. wi pm oaYs zmp025 ~e TIT 00390 i CL p---- ue ant oF FERPEaPian 228 3 a wf 3 ya 4" * oos8 w Res EsceExit | Fi-Help | Crti-PeEdit peaks - me - 600391 Co nese2 EAP Fagi0n2 : I 3 4 bo v 3 STRIVE Ru pon tis) Dara Ser I 2423 Tine "798 00392  J J a 00393 || Catitras ton curve of 950 Piuaride L rena 0.000078 :|J/ Sires PR cS 00394 | 1995-03-29 12:46 Output of : 95032921 Software : version 6.1 c1990,93 operator + DoW Date of the Analysis : 1995-03-29 10:24 Analysis File Name : C:\SKALAR\DATA\950329A1 LATmoopwTsT61g33l20Aa5sH)ES Frero FClauloirbirdaetio1n.5order = Inverse Logarithm : s10pe fs = hE So xas-acd1 x1_ == ccoorrrreecctteedd vvaalluuee ooff tthhee scaomnpcleentration 1 Result = 108 5 I 5 = Slope of the electrode aa21.= -00..0000008070 a0 -1l21638 FClauloirbirdaetioLn order = 2 Correlation : r = 0.99645 Result = a2 * x +al * x + a0 aa2l = 00..0000000200 a0 = 0.00843 . sampler Diluter TNyupneber Sample Time +iSAL000 : 50 sec. WAarsh TTiimmee i1120 sseecc.. Tsapkeeciaulp :+ Nsoinnegle needle Height : 70 mm. ndeieldulteionHeiFgahcttor:: 8100 mm ReGislauntpiloen Volume P: 12.5 ml. Dilution runs : 1 sReeprrocfeisle : No . mr 00395 1995-03-29 12:46 Output of : 950329A1 Fluoride 1.5 SPiagtnhalnumtbyepre :: De3bubbled ' DesycsotleomrNumber :: Y0es dRieLsuatmeple `: NNoo idiGl Tohuretsphoultd:: 40095 Window event : OFf ss12 ssTTaannddaarrdd :: IIggnnoorree s534 ssTTaannddaarrdd :: IIggnnoorree 5556 ssTTaannddaarrdd :: Igno0r.e150 s587 ssTTaannddaarrdd :: 00..360000 85910 ssTTaannddaradr:d: 11..250000 ODridmeernsi: onInv: erPsPeM Logarithm tstrairgtgevralLuimeit :: 5108000 DSeUc Peak shape : Pointed : esNtdhrt iiggnnoorree :: 16020 SSeecc MFeialstuerre window :: N7o5% Rfeogrennulearatio: n : No output: HE.##E Fluoride L Psaitghnalnumtbyepre :: De0bubbled DesycsotleomrNumber :+ 0No diRezsuatmeple :`NNoo daiilG oTuhtrpeusthold :: 04095 Window event : Off 00396 1995-03-29 12:46 output of : 950329A1 s512 ssTTaannddaarrdd :: 00..001350 5s34 ssTTaannddaarrdd :: 00..006900 s565 ssTTaannddaarrdd :: 00..112500 5578 ssTTaannddaarrdd :: IIggnnoorree +55310 ssTTaannddaarrdd :: IIggnnoorree oDirmdeenrsi: on2 : PPM sCtrairgtgervalLuiemit :: 5108000 DSUec psteAarktshiagpneore eNd ignore :: P60ointeSedc : 120 Sec . FMielatseurre window :: N7o5% Rfeogremnuelraatio: nc4:=:c3No output: H.H#H# 00397 1995-03-29 12:46 OutPut of : 950329A1 FFlluuoorriiddee L1.5 Ppeo Pos Typ Ident Dil Weight Ch Result F Cor. Valu Time wt iw Initial Wash 1 1.000 33 0.00.008621 0o 216To 6sTo 1c Tracer 1 1.000 33 1713.20079 22008688 22850 208 2a Drift 1 1.000 43 115.12304 22000911 23080 382 3 ow Wash 1 1.000 430.00.008681 o0 218To ss5To 4 si Standar1d 1 1.000 43 0.00.006625 3377 258 7320 5s s2 standar2d 1 1.000 33 0.00.200783 s9s5 3200 901 6 s3 standar3d 1 1.000 33 0.00.507981 220088 4360 10860 7 sa standar4d 1 1.000 33 0.00.510122 332200 552o 12600 ss Standards 1 1.000 G3 0.02.1a386 442255 6600 1436 5 s6 standar6d 1 1.000 33 0.04.317502 448859 7280 1608 10 s7 standar7d 1 1.000 13 0.03.220973 88933 11360 1784 11 ss Standar8d 1 1.000 43 0.06.264063 11241155 16640 19570 12 89 Standar9d 1 1.000 33 1.11.425278 22008858 23420 2131 13 s10 Standard 10 1 1.000 33 1.1s.417s4 22330088 25700 23090 1a Drift 1 1.000 a3 11d.e2477 22:10033 23560 24830 15 ow Wash 1 1.000 330.06.008641 oo 256To26o63 Page 1 of 3 : TUSl.Tppm : Wi pps 00398 1995-03-29 12048 ouseur ot +sssBEST COPY AVAILABLE. ... . FFlluuoorriiddee 1L.5 : EoHw Pos Typ Ident 203 weight Ch Rem7iCotr. valu Timeo hirer: Sere 6 ou BiK1 1 1.000 33 0000.70009 a2s5s5 siz0 2629ow @RY,TaSST FU BLK 2 1 1.000 13 0.00.203700 e68s 3210 3010ost Red < 8 ou BLK3 1 1.000 d3 o0i.0s67 4488 3000 31850 mesb. 1 uw sk 1 1.000 3 0.086 30.0503 180 180 432 3357 0 oh asked. -| 20 u sek 2 1 1.000 33 o00%.s0i8s7 e18e6 436o35o33t mss 2 uw sek 3 1 1.000 3Pos0a.0s8s5 117733 4220 3711ot wees 2u Fs2891-1 1 1.000 3 Absen A 13 260 3883 4 0.0111 13 0 0 0333 Nb 23 u Fs2e91-2 1 1.000 d3 oAbosemn A 1133 258o4o058 0333SD 2 u Fs2891-3 1 1.000 3 0.064 28 272 4231 4 0.0142 28 o o ~0426 = Wh 25 u Fs2852-1 1 1.000 3 0.067 49 292 4410 we 4 0.0188 49 0 o 26 a Drift 1 1.000 i3 ai1e.2s5s2 22010007 23480 4584 Mug. 27 w Wash 1 1.000 34 0.00.008631 0o 200o4t74e7 28 u Fs2965-1 1 1.000 23 001.606s6 4400 2800 49380 deseN . 25 u Fs2965-3 1 1.000 3 0.065 4 0.0155 34 274 510 34 0 TOYS: " 30 a Drift 1 1.000 13 11.2263 2211188 2358o5o204 rvagpe aw Wash 1 1.000 33 00..0006814 0o 240osoass 00399 1995-03-29 12:46 Output of : 950329A1 FFlluuoorriiddee L1.5 PPooyy Pos Typ Ident Dil Weight Ch Result F Cor. Valu Time wt rw RunOut Wash 1 1.000 33 0.00.008611 0o 256To575o9 Page 3 of 3 00400 2 me ante or o500200 Fiumrid 1.8 . dar wp3a u . " 7 $4 . s AN AR Raz me ETF TST 00401 BEST COPY AVAILABLE 600402 "  i 00403 Le 1995-03-30 10:59 output of : $50330A1 Software : version 6.1 1990,93 operator + oo Date of the Analysis : 1995-03-30 08:43 Oo Thzlas AWnweo 61332495S4)33 usm Analysis File Name : C:\SKALAR\DATA\950330A1 FClauloirbirdaetio1n.5order = Inverse Logarithm slope sn pobEE 6o xGaa-scas1> Result 108 si xG1 a == ScCoolrrorrpeeeccttoeefddtvhveaalluueeelecootffrottdhheee csaomnpcleentration 1 aa220i.:= 0o.l0o0o0o0s0s liza FClauloirbirdaetion order = 2 - Correlation : x = 0.99761 Resu=lat2 + x + al * x + 20 a22201.= 0oo.ll0oo0ot0oe0ss0sz sampler TNyupmeber Sample Tine +5irs0a100s0ec. WATazas.khe upTTiimmee 1120 sseecc. single apfeeecdilaelHeight : N7o0nema. Diluter nGGeiiellduultteiiooHnneViFogalhcuttmoer :: : 8100 2.5 mm ml. . RDeislaantpiioen runs i: i1 URseeprrocfeisle : No xr 00405 ` 1995-03-30 10:59 output of + 950330A1 Fluoride 1.5 BiPeaolcnohalloprumTsyepre |: De3 bubbled Yes fSeytssuateemsmgieNumber Nv0oo fG$ii3ondToouhntrpeBasucheeon:ld ||10o4t0e9s 22s13 SSsrTTaaannndddaaarrrddd ||: IIIgggnnnooorrreee 2251 SSrraannddaarrdd :| Izgnnoorree 223865 oSrstarTamaanannddaadarrraddd :i 0001....31200500000 - SD2r3id0meern"SsTtiaonnTdaav:redrPsMe: Loga1r.5i0t0hm StSeeaaisifiesoehrvaaplLeu'iemi|t |: s2p00o00i0ntSDeeUdc MSeeaaavseureliagnwnoiornreedow |1 7610%5%SSeece RFTieolgrteentrearation rNee Erte Fluoride L DSPeiadgtonhtaolnru.mtbyepre i:: Deebubbled RlSeyiseacrneepmieNumber : NN0oo . W3Sii5ndooTuwhrrpevasechnotld ||10o40f9s 00406 La 1995-03-30 10:59 Output of : 950330A s512 ssTTaannddaarrdd :: 00..001350 s534 ssTTaannddaarrdd :: 00..006300 s56s ssTTaannddaarrdd :: 00..112500 5578 ssTTaannddaarrdd :: IIggnnoorree . 55510 ssTTaannddaarrdd :: IIggnnoorree oDridmeernsi: on2 : PEM sttrairgtgervalLuiemit :: S180000 SDeUc psteaarktshiagpneore : P60ointeSedc eMNedasureigwnionredow :: 71205%Sec FRielgteenreration :: NNoe ofourmtuplaut:: c#4.:4=#c#3H 00407 1995-03-30 10:59 Output of : 950330A1 FFlluuoorridiedLe1.5 pPoou Pos Typ Ident Dil Weight Ch Result Cor. Valu Time wt iw Initial Wash 1 1.000 13 0.00.108785 o 0 o 15s 65oo 1 Tracer 1 1.000 3 1.248 2101 2274 211 4 b.8362 2100 o 2a Drift 1 1.000 43 0.18.523739 22113399 23290 3860 3 ow Wash 1 1.000 43 0.00.108745 o0 208"0 569To 4 sl Standar1d 1 1.000 33 00..0017850 oo 209To 755To 5 s2 standar2d 1 1.000 43 0.00.2067a9 2244 2340 906 6 83 standar3d 1 1.000 43 0.00.80995 112244 3350 10840 7 sa standar4d 1 1.000 33 0.00.818123 220077 4200 12620 8 ss standar5d 1 1.000 i3 002.1837 331133 5280 1435 5 s6 standar6d 1 1.000 43 0.04.17524 337177 5940 16120 10 87 Standar7d 1 1.000 43 0.02.823857 775533 9760 17850 11 ss standar8d 1 1.000 23 0.04.690%6 11330088 15400 19620 12 s9 Standar3d 1 1.000 43 0.18.322472 22000933 23380 21370 13 s10 Standard 10 1 1.000 330.15.644651 22338833 26360 23120 4a Drift 1 1.000 d3 o.1a.638066 22117733 23920 2487 15 Ww Wash 1 1.000 33 0.06.108785 o0 220o2e 729 Page 1 of 3 . . - 00408 1995-03-30 10:59 OutPut of : 9503301 FFlluuoorriiddee L1.5 Epogy Pos Typ Ident Dil Weight Ch Result Cor. Valu Time 16 ou blk1 1 1.000 230.00.200746 66 224o2o 022 17 u blk2 1 1.000 43 0.00.06714 7-7 2080 3013 18 ou plk3 1 1.000 a3 o00.m07s2 2-121 1920 31830 1 ou spk1 1 1.000 33 0.00.507925 111166 3280 3363 20 u spk2 1 1.000 43 0.00.500351 696 304oo 3536 2 wu spk3 1 1.000 340.00.803933 110066 3120 37060 2u spk4 1 1.000 43 0.00.600967 112266 3300 38880 23 u spk5 1 1.000 43 0.00.602938 113311 3320 40630 2 wu spk6 1 1.000 23 0.00.08986 112211 3200 42350 25 wu Fs2674-1 1 1.000 43 000.20727 1111 208To 4410 2a Drift 1 1.000 33 0.18.425665 22112222 23160 45870 27 w Wash 1 1.000 340.00.108745 o 0 1o 952 4o 766 28 u F52874-2 1 1.000 33 0.00.200746 &6 196o49o35 25 wu Fs2874-3 1 1.000 33 0.00.107715 -o 4 1o 84 o 5116 30 u Ps2879-1 1 1.000 43 0.00.01704 --1100 176 52740 31 ow Fs2679-2 1 1.000 i3 o0.00.103713 1-616 1630 S464 Page 2 of 3 . 00409 1995-03-30 10:59 Output of : 950330A1 FFlluuoorriiddee L1.5 PPoeyx Pos Typ Ident Dil Weight Ch Result Cor. Valu Time 32 wu Fs2889-1 1 1.000 33 0.00.404888 778 261o5o 640 33 ou Ps2889-2 1 1.000 G3 o00s.00901 9s5s 2766 5812 3a ou Ps2889-3 1 1.000 340.00.12232 224488 4260 59900 EE Fs2894-1 1 1.000 s3 0.00.s00981 9977 274o6o 164 36 u Ps2894-2 1 1.000 3Go0.00.s06924 111133 2880 6340 EL Ps2894-3 1 1.000 33 0.00.406849 884s 2580 6514 3a Drife 1 1.000 33 0.18.521756 2211336 23080 66820 39 Ww Wash 1 1.000 33 00..0017854 o0 170T6o87o2 40 u Ps2895-1 1 1.000 43 0.00.301841 3335 2080 7034 au Fs2895-2 1 1.000 33 0.00.301872 4400 2080 72120 FE Ps2895-3 1 1.000 33 0.00.202777 1133 1800 73880 43 a Drift 1 1.000 43 0.18.37180 22117788 23440 7565 aw Wash 1 1.000 33 00..0017858 00 165o77o96 we rw RunOut Wash 1 1.000 43 0.00.108745 0o 138o8o 040 Page 3 of 3 : 004i0 BEST COPY AVAILABLE oy av 3 3 wd SSN . was T o T TSG Tine Ed +Quests 06411 3 REST COPY AVAILABLE na ante or p03: Fivarice +5= 3 a "% xz | | EAN DETR of 9 Vga n woNS E T Te Tine e798 00412 ' 4000 ERTnau dCate7of 9W503I00Z81 E: FivIorTidNe 1.E5A) . 3 i $a wl \ qu Commei7t98 Fie | GreTPeaIt pea Tine . 11173 000413 cxtiaration curve of Soossows : Fivaride 1.8 sawn me Order : Inverseropmritha red =: 4921 000 000414 .