Document km96KM2zdmJrDk0J9jj671pvb

ARHe -0I5% 3M Environmental Laboratory Final Report- Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T-6053 in Rabbits In-Vivo Study Reference Number: HWI#6329-137 Study Number: AMDT-022195.1 Test Substance: FC-99 (T-6053) 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 and Revisions: AMDT-M-1-0, DThoehrrmmaalnEnxDtrXa2ct0i0o0n oOfrgFalnuiorciHdaelbiydeMAenaanlsyzoefr-aLMiovdeirfied AMDT-M-2-0, Fluoride Measurement Ton Analyzer by Means of an Orion EA940 Expandable AMDT-M-4-0, AMDT-M-5-0, Extraction of Fluorochemicals from Rabbit Liver Analysis of Rabbit Liver Extract for Fluorochemicals Using AMDT-M-8-0, Electrospray Mass Spectrometry Analysis of Fluoride Using the Skalar Segmented Flow Analyzer with Ton Selective Electrode Initiation Date: See attached protocol Author: James D. Johnson Approved By; he mes D. Jgfyfson tudy Dirofor 1/21% 2s Completion Date 03851 00wP0eo0sU1 1 tcAooftltalelercotdreegdarnm(iaHclWfIal#pup6olr3ii2cn9ea-.ti1o3Tn7h)oe.frFeTsCuh-le9ts9lio(vfTer-th6ai0ts523a8)n,adltayisysisssuwesahasonwdceosdmebrnuuosmtdeisdfafmaeprnledenscaenwsaelbryeeztewdoofnor the controls and with 250 mg/kg (trTe-a6t0e5d3)r.abbTihtsisinfcolrumduilnagtitohne ihsigahddioestehawnhoilcahmiwneersealrtabobfits treated spoelruftliuoonrotohcattanises0u.l0fo4n%atoef,FCT-w9e9ntsyolipdesr.ceTntheofdtohseesefosroltihdeshairgehtghreoCu8p,waTs6053 is a l2i0miutgsofkfgtwhietmhertehspoedctevteonCi8faplelrfolfutohreooscatmapnelseuslfwoenraetea.bsTohrbeedd.oseQ was below detection aTbhsiosrspttuidoyn owfasFCp-e9r9f.orDmaetdainfroormdeortthoerprfoovrimdseodfatpaerffolrutohreooacstsaensessmuelnftonoaftede(rFmCa.l95, wpthhoeaurlpmodatcbaoesksaiinuceomtnisvcaelnts)iteuwndetyre(maHarWvkIaei#lr6a3fbo2lre9-F(1CH3-W69I)9#6hi3fa2dt9hs-eh1d3oo0ws,nestHahWadItm#ip6ne3irs2ftl9eu-ro1er5do9ow)cet.arneAeshuilgfhonate enough. 3.1 Tes3t.,1.C1onAtnraolly,tiacnadl Reference Reference Substances Substance: and Matrices FC-95, lot 161 or 171. They are equivalent. 3.1.2 3.1.3 AAnnaallyyttiiccaall RCoenfterroelncSeuMbasttarnicxe::BNovoinnee liver and bovine serum 3.1.4 Analytical Control Matrix: Bovine liver and bovine serum g3.r2ocSeoruyrscteoreo,fbMoavtienreiaslesr:u3mMfrIoCmPS/iPgCmPaDCihviesmiiocnalfoCroFmCp-a9n5,y.bovine liver from 3.3. Purity and Strength of Reference Substance: Responsibility of Sponsor 3.4 Stability of Reference Substance: To be determined by Sponsor 3.5 For SbitoolroaggiecaClosnadmiptlieosnsthfeosrtToerasgteMiast-e2r0ia1ls0: CR.oom temperature for FC-95. 03852 V0000E 3.6 Disposition of Specimens: Biological tissues and fluids will be retainedper GLP Regulation for the time period required for studies longer then 28 days. 4.0 EXPERIMENTAL - Overview Tissue and serum samples collected samples were available for analysis at 28 days post dose were analyzed from HWI#6329-137. Liver for total organic fluorine by dceormmbaulstaibosnorapntidosne.lective ion electrode. The data were used to assess the extent of 5.0 EXPERIMENTAL - METHODS. 5.1 AMDT-M-1-0, Dohrmann DX2000 Thermal Organic Extraction of Fluoride Halide Analyzer-Liver by Meansof a 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 5E.l4ecAtrMoDsTpr-aMy-M5a-s0s,SApneacltyrsoimseotrfyRabbit Liver Extract for FluorochemicalsUsing 5.5 AMDT-M-8-0, Analysisof with Ion Selective Electrode Fluoride Using the Skalar Segmented Flow Analyzer The data from combustion and subsequent combustion analysis for total organic dfaltuaorifnreoomflliivveerrs saatm2p8ledsayfsropomsrtadbboistes sdhoosewdndoedrimfaflelryenwcietshbFeCt-w9e9enisdoasttiancghegdr.oupTsh.e Rabbits were dosed at 0, 10, 125, 250 mg/kg (0, and about 0.8, 10, and 20 ug/kg dwiietthhraensopleacmtitnoe tshaeltCo8f ppeerrfflluuoorrooooccttaanneessuullffoonnaattee).wiTthheafboorumtu2la5t%iosnoloifdsF.C-T9w9einstay pdeorsceenfotrotfhethheisgehsgolriodusparweasth2e0C8u.g/kTg-6w0i5th3 riessape0c.t0t4o%thseolpuetrifolnuoorfoFocCt-a9n9essuollfiodnsa.teT.he i(InnHtWprIra#ev6vei3no2ouu9ss-1sdt3ou0sd)ei,es(,aHnWFdIC#-s6u93i5t2a9hb-als1e5ab9se)ea,nmnsaohrtokdewernrmttoaolablseysepasebsrssdoiersrtbemenadtlaitanbdrsoaosbrbepisttisoofanf0t.e3r0amng/0kg3853 000603 3 dc(eoHrnWmcIal#lu6d3ae2bds9o-trh1ap5tt9ip)oe.nrfolIfunoFarCo-po9hc9at.ramnaeScsiuonlkcfieonnFeatCti-ec9w9sotcuuoldndytaboiefnsFaCpu-es9ref9fluul(oHmrWaoIor#ck6tea3rn2ef9so-ur1la3fso6sn)ea,stseimttewhneatsof csoanmseisatsenFtCt-h9at5nboutdejursmtaal daibfsfoerrpetnitosnalatpfpeoarrms(tFoCo-c9c5urisatthtehpesoetadsossieumlesvaellts)o, fit Ti-s6053, bHeoewnevaebrle,itfo all of the detect it. 20 ug/kg dose Since T-6053 were is the asbasmorebperde,ptahreatmieonthoofdFwCo-u9l9dunsoetd have in the pharmacokinetic study, a direct comparison is appropriate. Ostehgemrendtaetda was flow collected analyzer wuistihngfoenlescetlreocstpirvaeyemleacstsrosdpee(cstereomaeptpreyndaincdesS)k.alTahris data, acolntchlouusgihosnusptpaotretdivhee,reinanthdetohpeirenfioorne of is the not SdtiuscduysDsierdeicntodertaiisln.ot required to reach the t6h.a1tCmiarycuamfsfetcatntcheiss dtahtaataMnaalyysiAsffiesctthatthae lQounagleirttyeorfmtphhearDmaatcao:kiTnheeticcisrcstuumdsytaonncetshis s(abletlfoowrdmetoefcpteirofnlluiomriotso)c.taHneoswuelvfeorna,teit wsaesemnsotunpleirkfeolrymtehda.t The there dwoosuelwdabse very a low swuibtshtarnetsipaelctditfofedreepnocseitiinotnhienplhiavrerm.acokineticsoftwo salt forms at 28 days especially There levels does at 20 nuogt/kagp.peTahretodobseedweramsablealboswortphteidoentoefctFiCon-9l9imi(tTo-f60t5h3e)maettdheordm.al dose 8M .0 AIN OFT RAWDE ATAN ANDRA ECORN DS CE a8p.1proRpariwatDeastpaecainmdenDsa,taan:d Raw data, electronic daaptparwoivleldbpermotaoicnotla,inaepdprionvtehde final report, AMDT archives. 9.1 Protocol and Amendments A9.b1s.o1rpPtriootno/cToolxiacnidtyFiSnatludRyepoofTrt-:6H0W5I3#6in32Ra9b-b1i3t7s""S(iPnrgolteoc-oDlosteypDeerTmPa3l016.AB for dosing of animals, tissue collection, etc.) 9.1.2 Analytical protocol AMDT-022195.1 03854 000604 4 9.2 Signed Reports from Individual Scientists: None 9.3 Quality Assurance Unit Statement: See attached 9.4 Key Personnel Involved in the Study: See attached 9.5 Materials and Equipment: See methods 9.6 Solutions, Reagents, and Standards: See methods 9.7 Sample Preparation: See methods 9.8 Quality Control Practices: See methods 9.9 Test Methods: See Protocol AMDT-022195.1 9.10 Instrument Settings: See methods 9.11 Data: See attached. e9x.t1r1a.c1tiSounmfmoalrloyweadndbyraawnadlaytsai;s ug Fusing Oinriwohnolieonlaivnearlyazserd.etermined by thermal 9.11.2 Raw data; analysisofliver extracts using electrospray mass spectrometry. e9x.t1r1a.c3tiSounmfmoalrloyweadndbyraawnadlaytsai;s ug F" using Siknawlhaorlseeglimveenrtaesddfeltoewrmainnaeldyzbeyr thermal with ion selective electrode. 03855 000005 "9.S1i.n1glPer-oDtoocsoelDaenrdmaFlinAalbsRoerppotrito:n/HTWoIxi#c6i3t2y9S-t1u3d7y of dTo-s6i0n5g3oifannRiambablist,s"ti(sPsruoetcooclollecttyipoen,TePt-c.3)016.AB for 03856 0006U6 WISCONSIN SHIERA IAA HH Jp Sponsor: 3M Toxicology Service Medical Department St. Paul, Minnesota -- . EN - FINAL REPORT Study Title: Single-Dose Dermal Absorption/Toxicity Study of T-6053 in Rabbits Author: Steven M. Glaza Study Completion ate: July 11, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project dentification: HWI 6329-137 CO3857 Phone 508.241.0071 $a 1 of 42 EXPRESSMAIL DIAVERY 3301 Kimswan aivo Fax 000007 5032417727 wapisow. ay saris Page 2 of 45 HWI 6329-137 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Wisconsin, Inc., in accordance with the Food Assurance Unit of Hazleton and Drug Administrati (FDA) Good Laboratory Practice The following inspections were Regulations, 21 CFR 58.35 (b) conducted and findings report (6) t on (7). saSnttduadnydfairnDddiirneogpcsetroaratrieanngdismpsrauonecdaegdetumoreenHsta..zletWornittmeannagsetmaetnuts mroenptohrltys aocfcoirndesidpnegctoitootnhse FIrnsopmection DTaotes 12/21/94 01/25/95 01/30/95 02/10/95 04/10/95 04/20/95 04/20/95 07/10/95 12/21/94 01/26/95 01/30/95 02/10/95 04/10/95 04/20/95 04/20/95 07/10/95 Phase Protocol Review Protocol Amendment Protocol Amendment Necropsy Protocol Amendment Data/Report Review Data Review Report Rereview Date StRuedpyortDeidrecttoor MaDnaatgeemteont 12/22/94 01/26/95 01/30/95 02/10/95 04/10/95 04/20/95 04/20/95 07/10/95 01/10/95 02/10/95 02/10/95 03/10/95 05/10/95 05/10/95 05/10/95 08/10/95 Lu Ao Representative, Quality Assurance Unit 77.95 Date 03858 0000VS Page 3 of 45 STUDY IDENTIFICATION Single SDtousdey oDfermTa-l605A3bsoirnptRiaobnbi/tTsoxicity HT 6329-137 Test Material Sponsor Sponsor"s Representative Study Director Study Locations T-6053 T3oMxicology Service 3MMeCdeintcearl, DBelpdagr.tm2en2t0-2-02 SPt..0. PaBuolx, 3M3N22055133-3220 J3Mohn L. Butenhoff, PhD ToMxeidcioclaolgyDeSpearrvtimceent P3.M0.CenBtoxer,332B2l0dg. 220-2-02 S(t6.12)Pau7l3,3-1M9N62 55133-3220 HSatzelveetnonM.WiGslcaoznasin, Inc. MPa.d0.isoBno,x W7I54553707-7545 (608) 241-7292 InHiatizalletoTnestW:isconsin, Inc. 3B8u0i2ldPiancgkeNro.s 3 Avenue Madison, WI" 53704 ReHpalzalceetmoenntWiAsncimoanls:in, Inc. M3a3d01isoKni,nsmWIan B53o7u0l4evard StuStduydyTimIentiatbilaetion Date . EInx-pTeirfiemenEtnadlDa(tIen-1ife) Start Date SEtxupdeyrimCeonmtpalletiToenrmiDnaatetion Date JDaencueamrbyer133,0, 199I8o JFuelbyrua11r,y 1179,85 1995 July 11; 1985 y03859 0006VY Acute Toxicology SSttuedvyenDiM.recGtloarza Manager SFtruadnyciCsoo(rBduid)natW.orMcDonald PIant-riifceiaSuPpaedrgvhiasmor RReopsoertM. SBurpiedrvgiesor ToxicologySupport MaKnatahgyerMyers CSaulpveirnvisLo. r Horton Page 4 of 45 KEY PERSONNEL HHT 6329-137 Quality Assurance MSahneargreyrR. W. Petsel aboratory Aninal Medicine CDiipnldoymaJt.e,CarAyC,LAMDVM Supervisor Anatonical Patholoay ATnhaotmoamsicEa.l PaPlamtehro,logPihsDt DJeabcokraSherLf.orPti/rkel NSeucpreorpvsiysors ASnunpeervHiossohrer Pathology Data 03860 000010 Page 5 of 45 CONTENTS QStuuadlyityIdAesnstuirfiacnacteioSntatement KSeumymaPreyrsonnel ObRejgeucltaitvoeryComp]iance TTeesstt SaynsdteCmontrol Materials PRreoscueldtusres DSiigsncautsusrieon RePaftehroelnocgey Report Table 21 IInnddiivviidduuaall CalnidniMceaaln BSiogdnysWeights (g) 34 IInnddiivviidduuaall DPeartmhaollogIyrrCiotmamteinotnsScores 5 Individual Animal Tissue Weights and Bile Volumes AppPernotdoicxolA Deviations PPrroottooccooll ATmPe3n0d1m6e.nAtB No. 1 Protocol Amendment No. 2 HHI 6329-137 Page 32 46 88 8 109 1143 14 1154 1167 2139 2% 2287 4219 a" C03861 000011 Page 6 of 45 HHI 6329-137 SUMMARY Tskhiins sitrurdiytanwcays doofneT-6t0o53aswsheesns tahpepliseydstetomictheabsskoirnptoifonr/atbobxiitcsi.ty and relative TohfethsetuHdrya:wa(sNZWc)oSnPdFuctsetdraiunsinfgorthearcehe mtraelaetmaenndttghrroeuep.female acclimated rabbits Group Test Material Do_(snea/kLa)evel MNaulmebser of AFneimmaalelss 1 (Co2ntrol) Dis1t-i6l0l5e3d water 1o0 3 1-6053 125 33 3 33 3 4 T-6053 250 3 3 a* rAOnedepmliaancnieismdtaelwrietdshacaatrniofatihcdeeordseofnevmoaDllaueymea2noidfmuae2l..0tomlp/oksgs.ible broken back and orTfheestphebecatcirkavbeobfimtaesta.ecrhiaTrlhaebabtittretahwteanseincntldiipscpiatetedesd fdrroeesmeeaionfleedvhealiinrtwaaacsntd.ada mTishnieinsgaltreeeraeddoosfetoaotpfhpe)tihcecsktiinon dwgravesesrsncironavgpepreefddorwwiiatthh24-SahaorguaarnuzWeexrpabopasnudraaegndepeEsrelicaousdrt.eodplwaistth tpaappeertotapperovairdoeundanalolccleudsgievesng odbolshieonruivrcasatliaofontbsesrearnvtdaesttitowniocsrecwoeanrtderaoyclomnomdaruttceatrleiidatlyprceahddeomcsikensisatwnredarteaitocnoa.npdpurcAotdxedidimtaidtoaenilglyly 1,tlhie2n.rj5e,ecae)taenrd fpiunor-rploi2sf8eesd,atyesrb.meifonraBetoidoytneswte(iDoagyhrts2c9o)nw.terroelThdeematteiernrimitiainaleldaddoenmrimnDaialsytri-ar9triifotonartir(oaDnnaydorem1)ai.dziuntagindeswaaet mide 3bJ0oeafmdoiirnnegu)tt.eesstSafuotbresrecqoubnetanrnotdlagreemaatdreiermnoigvasallofad(dDmeairynmia2sl)traaitnrdiroinotnat(iDroaeyncsowre4dreeadndmaas8d.ethesBplpDoraooydxi1 nsaatnepllyes wIaenndrieti22ac.toilolneIcnt(eaDddadyiftr1i)oo,nn,aap`mpaartrogtxihinemaaltteilneyearo2f4v-ehninoeucrrosofpsptyhoesotndaonsDieamyal(s2D9a,ybefa2)op,rperoonxeixmpbaaetyresilmye4n,t2a08l,'nl15, tsoeferpmabirlnaoatotedidownaisnotfoobttsaheeirnuenidn-a1nfidrfoecnelpehlaaucslhearaannfidrmaalcn.teicornoAspl.sliesdAa.lmlpleTashneimwwaehlroselewceorlneitvreierfu,utghbeadniilsaeeoda at 2o{S5nev0ecotlmmiugaom/nelkegoonfalnydthdeoentedeerrfmmeiannlaeldeapfpionlriecaabictlhieo)ng.rosuipTtheewefrtreiosnscuoaellsllecfatrneoidmmalatsth,enefacnedrmaolpbesoytdhoasnkediddwnoeeiiygshefdromGO38E6: b$iilnto-eoTsdi,fesaapnmhdpalaseknesid.dns(easycesrriuwnfeircaeenddseocnnetllDuafylraozr2e'nwfertraoectictohonelsl)eS,cptoneTsdiovrebrusta,ftweebrirleet,enrotmdiernwmeaiatglihoenad.ppoflicTsahheteion 000012 Page 7 of 45 HWI 6329-137 WAbioptdphyliwtcehaietgihoetnxcegopafitniT.o-n605"oA3fll doniaedninGnarolotsupraep4spuelftaemraeildne calanniyinmitacelasltltymhaatneorrmiaall-rtehlraoutgehdoucthantgheesstiundy due to an study and tihnejurryepl(apcoesmseinbtleanbirmoaklenapbapceka)r.ed dernal irritation was This animatl wwaass sarcerpilfaceeeddinonthDeay normal throughout the study. No 2 the application of distoiblsleerdvewdataetr tohre were no test material-related lesions dermal T-6053 astcoarniyngofinttheervadlosse aslevaelrse.sultTheorfe observed at necropsy. 03863 000013 Page 8 of 45 HHI 6329-137 OBJECTIVE Trheelatoibvjeectskiivne rabbits. oifrrtihtiasncystoufdyawatsesttomaatsesreisasl twhheensysatpepmliicedtotoxicthiety/opaibesornpttion and REGULATORY COMPLIANCE PALRaaditbmbeoisrrniaisatstloturrdyaytiSwtoaunsd'isceosGn,odoud2c1teLCdaFbRoirn5a8ta,ocrcwyoirtdPharnatccheteicweeixtchReepgtthuieloantUi.Sot.nhsatFofooadrnalNayonsndicsiDirsuoigfcahle test tihhnoemotoghpieisnneiisottmnyuidxsytoulrauerbeiplirinetpyca,ormepadlnidfaonrscteatbhweiiltiGhtryoutphweas2AnnaionmtiamlcaolnWsdeulfcfotarerdec.oAncctAeRTne]tqrauptlrioeocsne.jduorne:s us`yeod duplicate anyofprtehveioSupsonwsoorrk.and study director, the study did notunnecessarily Identification TEST AND CONTROL MATERIALS cTlliheaeaurit,de.stcolmToahrteleercsiosanltrlowilaqusimdai.tdeernitailfiweadsadsisTt-i6l0l5e3d awnadtedrescanrdibewads adsesacrcilbeaanr', accoslorless Purity and Stability PdIaehtteeerrSimpaiolnnasmtoiiroxntasusrseu(miepnsrcelpuraderisenpdgonsfuoinrbdietrlhiettyeGstrfooucrpontd2eistatinoimnmasat)el.rsiafloArnaplcuyorsniictesyntaorfnadtstihtoeanb.viuleity btSeopmaoodgoeenqreuiattyet/esfoolrPuubrtihiletiytpyua,rnpdoassnedtsabsiotlfaibttiyhliistoyfsttwuahdsey.cnoonttrcoolndmuactteerdiaolr wreerqeuecsotnesdidbayretnheto Storage and Retention yTsfihartemehpslztHeeearsztolfseeattenoadcthocomnttaerisotnltaainmndateacronittaerlmosplerwaemtarutereersitaoolfrewd2a0satCtar+ko1eo0nn tanefodmrp"ewir1la0ltuyerbaeer.s.reAtinairmneeesqveorrvinceacace LtaessttinmgatearcicaolnrdiwWnaigssctoronestHiuHnrIne(SdHOHPIt.)o other testing StAhtneayndSaprroedmnasioOnrpienragafttcieornngtcroPomrlpolcemetadtiueorrneiao(lfSOPai)ls.i rieAntnyayiimannrifonr and will not be discarded after issuance of the fina] Tort 603864 000013 Page 9 of 45 Safety Precautions HHI 6329-137 Tahned pvoelsitciaensd. control material handling procedures were according to HMI S0ps Test Aninal TEST SYSTEM WyDDieeaspnrclvteoarcn,esbmieniPnetfnnanacsniyrillanivbatablyniitwasaatsoon3f80Dt2ehceePHanrcbake:err(s2V8,A)vSe1Pn9uE5e4,satMnradadiimnsaoiwnne,traeiWnipersdoccouanrtseidtnhyfrMtooancziHoirp,sn Inc., J33a0n1uaKriyns1n1a,n 1B9o9u5leavnadrdw,asrMeamcdaeiiisnvotenad,infeWridosncaotHnRsPti,hne.IHnac.z,letKoanlamWaizsocoo,nsiHnichfiaggesn].ioonn at Housing pphoaeuresierndg, raeIcnccelisipctnr.aeteinto-hnbeotaatnnodinnatlhssrtoauiwgnehlroeeustsacsctthleeienlasttucedadyg,efsotrhiena controlled quarters. paenriimoadls owferaet temperature: lIenadsitvid7ucdsayyss. and hunssiis- Jt1ha2ei-snhetoauircnon1diaigthtiteo/mnIps2e-rhaeoxtuiusrrteedEdaon,rfvkitr1hoT9eniymgehtwntoetiran2elg3Ccdc,yooccnlutaemr.eornletsleaIdntfiocvraestehhsuenwiahdneiirntedy] voafrraiSgssotiwscoconegs.rcoaitnn'.zo a adverse effect on the study outcome. and considered to have fiom Anima) Diet [aLnhaaebloyrazanetidomraylbysRtawhbeebrietmapnDruiofevaticdtHeuFdre#ar5c3c2fe6os,rs contaminants. Samples ntPHuoItwraFietteeidrosn,a]dInc1coi.mbpiotnTuehnnetasnfdeeadnad m1essnaysriuoerucetodinncearrmtsoaulnt of yienrteerfneorekdnowwinthconortamaifnfaencottfsedthinethewthaetreesrfueleatdrseoorpfewratihtoeedrisctaautldlyly.evaenlaslyztehadt bpy oMuIt. hahrere Selection of Test Aninals uwpTerehioreggehraatpnnli.avnacarelidTsahtewiienotrrnoaenodsifotdmueitdnzhyteaitgfiraiooneniudmpasbblyosduysainwniemgailgahtnssutmrwbaeetrriefiadeneddtebcroomdriyrneewsdepiognohndtinDgpuegaagr cteaeog`itiain:nnds btShoteduyd5yw%edpiirgdohbtnasobtifloeirxtcyeeeacdTheve2glr.osutpa"nOodnfearefdeafmcdoahrelveiesaeaxcatnhiiownnegasrrleouopnf(oNotot.hfe`Fse5tmaa3ect4ahi5ns4st)weiexciaglshletyj.adciiseufddretfnhotnretChocree"n 02865 COZ treesptlacmeadterwiiatlh eaxnoptohseurrefedumealetoanaimpaolssi(bNol.e bPr5o3k5e0n).back. Thiswasa'nroern))acceocs fice 000015 Page 10 of 45 Study Desian HHI 6329-137 Ainnitomaltshewefioglhlionwgingfrosntud2y,31g1routpos:2,609 g at initiation of treatment were placed Group Test Materia Domsae/kLa)evel MNaulmebser of FAenmiamlaelss 1 (Control) 2 DiTs-ti6l0l5e3d water 100 3 T-6053 125 33 3 33 3 4 T-6053 250 3 3 *3 rAOednpemlianacinesidmtaelrwietdshacaratinfoaitchdeeodrsefonevmoaDllaueymea2noidfmuae2l..0toamTlh/ikpsgo.ssaniibnlael bwreoikgehnedba2c,k38a7nd at initiation. JustifforiSpceca iestSei lecotinon cHhiositcoreicbaelclayu,setohfe Ntheew ZlaeragleandamoWuhnitteofalbbaicnkogrroaubnbditihnafsormbaeteniontheon atnhiym:alspeocfies. PROCEDURES PrepaofrEa xpotsuireo Arn ea TTOghhnheeytiahicienlipdupaneybdlebmeairfesoahreemdatdseeksitnu)pm,a'taeptrhpierao]xFilmaaanptkpeslliycoaft2i0eo%anc,hofrtthahebebibtatoctkwaalsandbc,oldiypipfesdunrefcFaercsaessarasryhenh(:otoor. asiinrtiremisat.lansttiaowcnet,resokrcilnidpeopffeedctthsoen ttDheaastytswso8iutleadnsd wpar2s9ecltiuondseapiedcthteeind uvsfeiosruoafilnitazneiyrnfgoefrtihnteghe aJpseenpilominmcia:st.ion The Dose Administration Amlalter2inailn.alsThreecdeaiyveodfatrseiantgmleentadwmaisnidsetsriagtniaotnedofastheDayres1.pective test or control So(rndoiusaptcihll1. eadnAinwmaatlie'nrsd)ivbiwoaddsuyalwaepipdgloihseted o(ne2v.e0tnhlemyl/dkatgyo)tohfweasttreecsaattlmceusnlitat.etedatThaeandvcaomtneetarsoou]fremdajbearsieod] approximately 0.05 ml/cnt. C03866 000016 Page 11 of 45 HHT 6329-137 G(rTo8u0p5s3)2. wa3.s applied ata mainxded4. witFhorditshteilGlreodup-w2atearnimtaolsa dose volume of 0.1 ml/kg. This c(o10ncemng/tkrga)t,iontheofte10s0t mmagt/emrLiaalnd mixture was stored at room u2t5ne0dmipnlegur/taketgdu,retroeusntpthieelcttieavsdetmliyns)iistteuessrienodgf.tthheeThaGervoeturepassgtem3abtuaelnrkdiad4lenasnwiaitsmyalasodfmi(0n1.i295s7t5eorrge/dnl to mdraeetsteperericmatilinveemlityxh)te.urdeoAsnweasvioncldaiulvmiceduulfaaoltreddeoasfcehorodfeoascethhelaenvreielmsaple(c0bt.ai1sv3eedatndeonst0.imt2sa6tebmroLid/akylg,weoirghtteston dtohsee dalyeveolfs application twraesatm4,ent1.6, ranged from aTnhde 2a5recan'o,f reexsppoescutrievefloyr. 0.02 to 0.06 ml/cn', thTehe10a,pp1r2o5x,imaantde 2r5a0tengo/fkg sEeaccuhreadrewaitohf Elastoplast ptaaappppeelrictoattapiproeonvairwdaoesunadcnovaloelcrceldeudswgieivstehadnadre1so0sv-iecnrngw.rxap1pC0eo-dlclnawrigstahuwzeeSraerbaansndeWadrgeapLo and restrain the animals during the 24-hour exposure period. Awrapespsrtorrxeamiionmviaentdgelwyciotl2hl4arthsaopuraswndataefbrtaenardnadgteesdsitswpeororesacbrolenetmroovpleadpemraatnedtroiwaaenllys.arepspildiucaaltiotne,st tmhaeterial ReasonforRoute ofAdministration The dermal route is a potential route of exposure in humans. Observations of Animals C4olbishneoirucvraasltiaofontbsesreravntdaetstitownioscrewcoaenrtedraoyclomnomdaruttceatrleiidatlyprceahddeomcsikensisatn(rdmaotraintoinna.gppraAnoddxdiimatfaittoeenrlanyloo1n,c)li2wn.ei5c,raeland conducted daily thereafter for 28 days. mBaotdeyriwaelighatdsninwiesrteradteitoenrmi(nDeady 1f)o,r ranadndoatmiziant-iloinfeputrepromsiensatioonnDa(ybay-9,39)b.efore test rTaehdaemdiininingsi)tt.iraaltiSoudnebrsmeaaqcluceonritdrirnrigetaadttioinogntsheroeDfardadiienzrgen!aylatseicmrharndiietqautbeieofn(orreweceortreedsemtdadoears actpohnpetrrooDxlaiynam1tateelryia3l0 minutes after bandage removal (Day 2) and on Days 4 and 8. Sample Collections 03867 B2b1ll1ooooddanwisamasamlpslceosnbdeuf(coatrpeepdreoxaxppiempraritomexelinymtaat4lelnyLi)n2iw4te-irhaeotuirocsnolpl(oeDscattydeod1s)e.fro(SmDauybasme2)aq,rugeinantnadlcoonelalreDacytvsieoinn4, oof8f, 15, and 22. In addition, at the time of necropsy on Day 29, approximately 000017 Page 12 of 45 2s0ampmliesofwebrleoodcewnatsrifoubgteadineadnd fsreopmartahteedpositnetoriosreruvnenaandcavcaellouflaerachfraacnHtiUimIoaln6.s3.29A-l1l37 T~h2e0sCe s1a0mpClesuntwielreshtihpepnedsttooredtheinSpaonfsroere.zer set. to maintain a temporatese of Pathology wAaintdthtneesrcomrdioinpuasntiiepodenntoionfbartrahbneidtoeanxlp,eorrdibemlree.ndtavlTihaepthhaseisteepso(sDotafeyrit2eo9)sr,t vaeannndaimcacoalnvsta,rwoelrexemsaaatnnegerusiitnnh]getteisz.ed Aaalnplyplaianbcinanotarilmosanliwwteeirreees wswauesbrhjeeecdtreewdciotrhtdoeTdau.nkewaTabhrbenrewvthiaopalteewdatlgiervreorsb,sefnboierlceer,otphaseysneeexccatrmioiopnnsaytoifporn.othceecdouyre. danedr,maflemaalpepliincateiaocnh with the exception gosfriotutephefwreaornniemaalcliollsaeanccitrmeaid.lf'si,cedTahneadndtbointsehscureKoipdssnaiemeypdslesofnrafDmoarythal2e,l Fainriemsalmea.le were wPleaicgehdedon(vdorlyuneiceonl(yinidteitaelrnitneesdt maintain a temperature of -20C af1no0irmCab.lisle)Ao.fnltye)rT,henetchsreaonmppslpyel,sacewtdheereinaniiammmafdlriseaeteseer]reyset to discarded. Shipment ile, and Tissues fljfcietv)eerrist,ontblihilefe,eSpodtenersrmomarilna(taJipaompnel,sicDat.thieJoonbhlnossoiotdne,ss,a3mMapnldeE.sEk.i(ds&neeryPus.mC.wa,enrdeBlcdsegel.nltu2lfa-rr3o0zfe9rna,ct(9io3on5nsd)Br,uysh Asvavomelpnuluneees,s.. St.HTAhIePauSdlop,eosnsWNno,ort5is5a1c0cr6ee)ps,tponaasnlyiobnlrgeeswpifotonhrsittbhhieeliirrteytceonrftorireosnpthoenadndainnagdliyswspeioissgihtotfisonotrh.eof the rteipsosrute. samples collected in this study nor are these results presented in this Statistical Analyses No statistical analyses were required by the protocol. LocationofRawData, Records.and Final Report Trheetairnaewddaitna,therecaorrcdhsi,vesanodfanHUIoriigninaaclcorsdiagnnceed wciotphy oHNfI thSeOP.final report will be 03868 000018 Page 13 of 45 Body Weights RESULTS wyeoidgihvtidugaalinsandfrommeanDaybod1ytwoeiDgayhts29.are in Table 1. HHI 6329-137 Al) aninals exhibited body Clinical Observations tIhnrdoiuvgidhuoault tchleinisctauldyswiigtnhs tahree finollToawbileng2.exceApl]tioann:inals appeared normal + TtOohnieshaGvarenoiumpianljuwrfaeesdmasliaetcsr(ibNfaoi.cckeFda5,t345nt4he)ecrottpirsneieaetdeo,df replacement aninal ap tawhniedthr2e4Tp--lh6ao0cu5er3d Bawltioto2hd50Ncoom.lgl/ekFgc53t5an0py4p.eareTdhe peared normal throughout the study. Dermal Irritation apInrniodmdiauvlicsdeudatlrnoeadtdeeerdrmmaalwlitihirrrTri-it6ta0at5tii3oonna.tscaonNryoesdoefarrmteahleindiorTsraebiltealtevi3.eolns.wTahse ccbosnterrovledmaitnertihael Pathology oGaInnrniodDmuiaapvlyid4u2t,aiflsesmruaeaelvneeimawale(leiNdgo.hptaasFtSfh3roa4aln5cdo4tg)yb,ilcetormemvaeotnletudstewsairteahrepTr-ei6ns0e5nT3taebdlate i25n.50TamTbghl/eekgn4.ecanrdoIpnssdyaicveiosdfufaihlreed Tesions observed in any ofurteheofremthaeinifnigrstaniTmuanlbsa.r vertebra. There wore ne Page 15 containsa pathology report by the study pathologist. 0389 00001Y Page 14 of 45 DISCUSSION HHT 6329-137 wdTeehrremeaalecvuaatplepulasitycesadtteimioinnc.maalbeTshoearrnpedtiwofenerm/eatloenxoicatiletbsiytnoamnadrtaebrrbeiiltaastiwvheenskiandniinrirsittearnccyy of as T-6053 a single mwaetiegrhitalg-arienlaotredind-elrimfael cliirnriictaaltioFninwdaisngsobsaetrvaendyldo-ufrreitlnhagetoddthoeccehsatnlugedveyes.lsi.n body No test SIGNATURE Be . WN SSttuedvyenDiW.recGtloarza Acute Toxicology BN 9 Date REFERENCE 1. oTDrofaxiitzcheie,tyS,aJ.fAestHsy.,ocoif"aAtCcihuoetnmeicoDaferlmsFaoloidnTFoaonxodidcsDi,rtuygDr(uOSfgifsnigclaienadlECxopsomseurtei)c,s" -DIen:rmaAlppraisal (1959). s of the U.S., pp. 54-56 0oJE70 000020 Page 15 of 45 HHT 6329-137 PATHOLOGY REPORT 2STTehhveoerhlees woeperfren0,iSniax1t0,iroanb1b2o5i,fts `tah(nedth2srt5eu0edymn.agl/ekOsgneoafndfebmotadhlyreeWee(iAfgnehimmtaallessu)Ntoh.eaanFciehsseefsreoc)nodndfeocurrdoopssieed etbaaa5ccs0hkk.mamaa/tkTehgriisoafla,nbionddaoylsewewailsgevhetrle,wpalsdaacyesdaocfwriidtfehiacteahdn,otohnaendrDagyfreom2saslbeeocba(sunesirenvean)tpiofenoos. arpF5pa3ano5re0eo4ny)t.'"osbrTohkeen ninal are in the Individual Pathology Coments that folten croonirdedpfoorrt. ASteudnyecrtoeprsmyi,nattihoenr.e mdaelrmeala application weThree site nfloriovenvri,esaicbbhlileeo,fletashnieosnaespparinonixmiaamnlayst.eofa1n-tdchme baxatnhiTnoaoknlisdnseseyaacrcihficceotd haet cTTohrlealsehLeisdnsdu(eovsnoelfumrfeoenmoanlthleey ifdneenteaealrcemhing(erddoousepfdorweatrbeil2e5c)0o,lmlaef/crktoaez)edn.,SacaTnrhdiefitsceienodtcnoetnosbatuhmgeple3sSseofawreneocsrmee.one cted but were not weighed. ~ After necropsy, the animels nerdiscarded. TThhies afniinnadlingsaccorrirfeilcaetdedonwiDtahy back. 2thehacdlianifcraalctoubrseerovfattihoen Foifrstan TsupnpbaarrenvsertLeoborkae.r PThaotmahsologi,st"Palnes', PI 0(460362995-137.51h) Date Zot F5" 03871 000021 Page 16 of 45 Individual and TMaebalne 1 Body Weights (g) HHT 6329-137 --_-- Animal Mumber oie F[SO4ASLL Mean [[2S48T3 Fo452 Mean [TaSsSe Fae Mean [[RNAsS F53439 Mean Randtoan-le ization Day -- RandFone-male Animal ization Day Day-9 TIT 75 Number Day-9 3 29 Group 1 (Control) - Distilled Water (0 ma/ko; 220 22,20885 2243183 320003 FmSuMsss 253 3088 FSM2 22043m7 2497 g2l,a5n7s 2 ae gasa Sos 2,249 2,442 3,004 2,408 2,512 2,072 Group 2 - T6053 (10 ma/kg 2222011 2,08 2z.84694 22,09m7 FFSsMMMM8 Z362 2777 FsMe0 224,7368 2,41 22,43s3 5gl,e7ees8 sls 5lers 2,178 2,455 2,897 2,415 2,447 2,905 Group 3 - T-6053 (125 ma/kg 220.6255 2.1% 22,,442957 22,,80643 FFSSEMT 2,33 243 FSO 224,7463 zr 2os,e5]0 25,080084 253 3008 2,232 2,415 2,850 2,486 2,554 2,037 Group 4 - T-6053 (250 ma/ka) 220920 323081 2277a% FFsSuSMs 22,,02907 22333080 52,960580 2,206 2,380 2,942 FF55334s5046 2.T4a24 22,365029 22,,963297 2,172 2,343 2,827 2,314 2,359 2,79 @NA ANnoitnaalvaiNlo.ablFe5.3454 was originally use in the study and was treated. seTlheicstedanibmyalthweasransdaocmriizfaicteidonafprroo.gran for creopmlpalceetdionwitohf Ntoh.e eF5x3p5o0s4u.re TpheeriobdodyduweeitgohtaspofosrsiNbol.e bFr5o3k4e5n4 abpaccknoarnd was b Dianycl2udebdodyinwetihgehtD.ay 1 or Day 29 group means. Cc03872 Replacement animal (not included in group assignment randomization). O00 "_ Page 17 of 45 Sex Male Female IndividualTabClleini2cal Signs Aninal 1-4 Hours Number _O_bservation (pay1) Group 1 (Control) - Distilled Water (0 F53464 Appeared normal v F53441 Appeared normal v F53451 Appeared normal v F53461 Appeared normal v F53455 Appeared normal v 53442 Appeared normal v HHI Day 2 3through ma/ka) v v v v / v v v v v v v 6329-137 29 Group 2 - T-6053 (10 ma/ka) Male F53447 Appeared normal F53463 Appeared normal F53452 Appeared normal Female F53444 Appeared normal F53448 Appeared normal F53460 Appeared normal vv v vv v vv v vv v vv v vv v 7 Condition existed. 03873 000023 Page 18 of 45 Sex Male Female Animal lumber F53453 F53458 F53446 F53443 F53467 F53450 IndiTvaibdlueal2 C(lCionnitcianluedS)igns 1-4 Hours _O_bservation (pay) Group 3 - T-6053 (125 mg/ka Appeared normal v Appeared normal v Appeared normal v Appeared normal v Appeared normal v Appeared normal v HAI 6329-137 Day -2. 3throug2h3 v v v v v v v v v v v v Group 4 - 7-60! Male F53459 Appeared normal vv v F5345 Appeared normal vv v F53439 Appeared normal vv v Female F53449 Appeared normal v/ v F53456 Appeared normal vv v F53454 APpopsesaibrleed nborromkaeln back v - b(laetedtiinnge oifnte2r4v-ahlo)ur - , Moribund sacrifice -7 F53504 Appeared normal /v v /- CCoonnddiittiioonn neoxtisteevdi.dent. a Animal No. F53504 replaced Animal No. F53454. 03874 000023 Page 19 of 45 Individual DermTaalbleIrr3itation Scores Group 1 (Control) - Distilled Water (0 mg/kg) HHI 6329-137 DermalReaction EErdyetnhaema ADetsoqnuiaanation CFoirsisaucreionugsness -- MStaudlyeDsay IT 2 4B Animal No. F53464 0 0 0 0 0 0 oo oo 00 00 00 0oo 00 00 00 o0o -- fSteumdayleDasy I aw Animal No. 5346 00 00 00 0oo 00 00 00 0 0o 00 00 00 EEdryetnhaema ADetsoqnuiaanation FiCsorsiuarcienogusness Aninal No. F5344 00 00 00 oooo 00 00 00 o0o 00 00 00 o0o Aninal No. F53455 0[I 0 0 0 00 00 00 00 00 00 00 00 EErdyetnhaena ADetsoqnuiaanat ion FiCsorsiuarcienogusness Animal No. F53451 00 00 00 oooo 00 00 00 0oo 00 00 00 oo Animal No. F53442 00 00 00 00 [0I 0 0 0 00 00 00 00 C0387S 000025 Page 20 of 45 Table 3 (Continued) Individual Dermal Irritation Scores HWI 6329-137 Group 2 - T-6053 (10mg/kg) ermalReaction-- --t IT Sa2dl v4Oeasy f"eSmtaubladyeys I 2 AF Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F53447 0 0 0 0 0 0 0 0 0 I] 0 0 0 0 0 0 0 I 0 0 0 0 0 0 Animal No. F53444 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 J Animal No. F53463 Animal No. F53448 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 0 0 0 0 0 [3 0 0 0 0 0 0 0 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 J 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. 0 [ 00 [3 0 0 0 0 0 0 0 F53452 0 0 00 0 0 0 0 0 0 0 0 Animal No. F53460 0 0 0 [J 0 0 0 0 [J 0 0 0 0 0 0 0 0 0 0 0 0 [ 0 0 03876 000026 Page 21 of 45 IndividuaTlablDeerm3al(CIorntriintuaetdi)on Scores HHI 6329-137 Group 3 - T-6053 (125 mg/kg) . -- SMtaudlyeDsay' Dermal Reaction TT 2 4 8 Animal No. F53453 Erythema Edema Atonia CDoersiqaucaemoautsinoenss Fissuring 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 [I] 00 0 oo Animal No. F53458 Erythema Edema Atonia DCeosrqiuaacmeaoutsinoenss Fissuring 0 [I] 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 o000 Animal No. F53446 Erythema Edema Atonia CDoersiqaucaemoautsinoenss Fissuring 0 0 0 0 0 0 0 0 0000 0 [I 0 0 0 0 0000 StFuedmyaleDsay I za 8 Animal No. F53443 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 90 0 0 0 0 0 oo 0 0 Animal No. F53467 0000 0 0 0 0 00 00 [I 0 0 0 0 0 0 0 0 oo Animal No. F53450 0 0 0 0 oo 0 0 0 0 0 0 0 0 0 0 0 0 oo 0 0 0 oo 0 0 03877 000027 Page 22 of 45 IndividuaTlablDeerm3al(CIorntriintuaetdi)on Scores Group 4 - T-6053 (250 mg/kg) HHI 6329-137 -- Males DermalReaction -- I S2tudyD4ay Animal No. F53459 EErdyetrhaena 00 00 00 o0o ADetsoqnuiaamation 00 00 00 0oo FiCsorsiuarcienogusness ~~ 00 00 00 o0o Aninal No. F53445 EErdyetmhaema ADetsoqnuiaanation CFoirsisaucreionugsness 00 00 00 o0o 0 0 0 0 0 0 0 0 00 00 00 00 Animal No. F53439 EErdyetmhaema ADetsoqnuiaamation CFoirsisaucreionugsness 00 00 00 oooo 00 00 00 o0o 00 00 00 00 Females I oStoudy Day AE Aninal No. Fs3449 00 00 00 00 0oo 00 00 o0o 00 00 00 00 Aninal No. F53456 00 00 00 00 00 00 00 00 00 00 00 0oo Aninal No. Fs3454 00 0 0. . o oo o. . == o oo o. . EErdyetnhaema ADetsoqnuiaamat on CFoirsisaucreionugsness Animal No. Fs3504 00 00 00 00 0 0 00 00 00 00 00 00 00 ba- ANRonetipmlaaalpcpelmrieecnpatlbalcaeen.dimawlithforNo.No.F53F5530445.4. C03878 . 000028 Page 23 of 45 Alnuimmbaelr F53464 F53441 F53451 F53461 F53455 F53442 Table 4 Individual Pathology Comments Sex Died TestSaDcariyficed NecropsyObservation Group ontrol) - Distilled Hater (0 mg/kg] M - 29 No visible lesions. M - 29 No visible lesions. M - 29 No visible lesions. F - 29 No visible Tesions. F - 29 No visible lesions. F - 29 No visible lesions. F53447 M - F53463 M - F53452 M - F53444 F - F53448 F - F53460 F - - Not applicable. roup 2 29 29 29 29 29 29 T-6053 (10 ma/kg] No visible lesions. No visible lesions. No visible lesions. No visible lesions. No visible lesions. No visible lesions. HWI 6329-137 03879 000029 Page 24 of 45 Animal Number F53453 53458 Fs3446 F53443 F53467 F53450 IndiviTdauballe 4Pat(hCoolnotgiynueCdo)mments HHT 6329-137 Test Day Sex. Died Sacrificed NecropsyObservation Group 3 - T-6053 (125 ma/ka "HM M- 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. F- 29 No visible lesions. F- 29 No visible lesions. F- 29 No visible lesions. F53459 WM 53445 M F53439 H F53449 F F53456 F F535 F F53508 F Group 4 - T-6053 (250 ma/ka - 29 No visible Tesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 29 No visible lesions. - 2 fTrhaectfuirresdt. lumbar vertebra is - 29 No visible lesions. 2- ANnoitmaalpplriecpalbalcee.d with No. F53504. b Replacement animal for No. F53454. 03880 000030 Se Male Female Male Female Page 25 of 45 Individual Animal Table 5 Tissue Weights and Bile Volumes HAI 6329-137 niet Weight ---- (g] vera Appr (Bite Group1 (Control) - Distilled Water (0 ma/ke F53464 87.407 - 0.468 0.3 F53441 114.827 16.722 0.331 2.5 F53451 89.123 - 0.394 0.3 F53461 73.175 - 0.371 0.7 F53455 94.647 16.740 0.512 Lo F53482 91.500 - 0.391 1.5 Group 2 - T-6053 (10 maka F53847 81.754 - 0.213 2 F53463 83.148 15.119 0.357 0.6 F53452 82.687 - 0.535 0.3 F53444 70.587 - 0.719 0.7 F53448 74.702 - 0.361 0.7 F53460 92.881 15.925 0.368 0.7 a NVooltumaepplnioctabmleea.sured to tenths of a mL. 03881 000031 Page 26 of 45 Sex Male Female HWI 6329-137 Table 5 (Continued) Individual Animal Tissue Weights and Bile Volumes Animal Mumber ---- Liver Weight(@) Dermal Appli- Kidneys cationSite GT roup- 3-60 (125 5 ma/3 kq) Bile Volume(mL) F53453 96.249 - 0.612 L5 F53458 77.755 - 0.390 0.5 F53446 83.574 17.429 0.321 0.7 F53443 79.767 - 0.602 0.6 F53467 78.684 15.937 0.467 0.7 F53450 79.782 - 0.580 2 Group 4 - T-6053 (250 mg/kg; Male F53459 79.255 15.567 0.372 0.4 F53445 97.381 - 0.270 0.5 F53439 83.548 - 0.349 0.5 Female F53449 73.983 - 0.391 0.4 F53456 77.804 14.412 0.572 1.0 F53454 NA - NA NA F53504 83.081 - 1.253 11 - Not applicable. a Volume not measured to tenths of a mL. Obs Rao repacedwith ho FLaLeReTt became effective Coassz b Moribund sacrifice on Day 2 (01/14/95) Tissues were collected, however, their due to possible broken back. corresponding weights or Replacement animal for No. F53454. - 000032 Page 27 of 45 APPENDIX A PPrroottooccooll DTePv3i0a1t6i.oAnBs PPrroottooccooll AAmmeennddmmeenntt No. No. 1 2 HHT 6329-137 03883 000033 Page 28 of 45 Protocol Deviations HUI 6329-137 Protoco CP.agOebs8e,rv7a.tiEoxnperoifmeAnntiamlalsD,esi(g4n), oSfampColleleCcotliloenc,tioSnesc,ond(c)SenMteetnhcoed. A(papcrtuoaxlimavtoelluyne 20tonLbe odfocbulmoeondted oinbtatihneedrawfrdoantat)he wiplolstebreior svaecnraifciacvead ofin eaa cmhorainbiumanld ctoinmdeitoifonnecorropssaycri(fDiacyed29)a.t the Actual Procedure. cBo1l0l0edctweads aitnatdhveerttiemnetloyfnot anneicmraolpsy(Nof.ronF53t4h5e4)fetmraelaeted at 2on50Danyg/k2gdutehattowaasn sapapcarriefnitced broken back. DP.age"Pa8t,ho7l.ogyE,xpe(r1)imeUnntsaclhedDuelsiegdn, SSeanctreinfciec.es AannidmaDlesathisn, aSecond maonreisbtuhnedtizceodndiwtiitohnswoidlilumbe tpheentombaarrgbiintalal ea(rvivaeiinn)j,ectbiloend in vexisaantghueinvaetnead.cava, and tThreeatfeedmalate a25n0imamlg/k(gNo.wasF53454) sapapcarriefnitcedbroonkenDaybac2kduuesintgo aann odivoexriedxep.osure to carbon DP.agePat9h,ol7o.gyE,xpe(r3)imeSnatmaplleDesign, TChoellewchtoiloen,ivSeerc,ondbilSee,nteannce. oafpptrhoexidmeartmeal1-acpmplxic1a-tcinonsecstiiteon kfirdonmeyasll fraoninmatlhse, fiarndstbmoatlhe and wfielmlalebenecoclrloepcstieedd ainndeach group siemtmedtoiamtaeilnytapilnaceadteinmpearaftruereezer of -20C 210C. Tahpeplisciazteioonf tsihteedecromlallected at dnoeccurmoepnstyedwasforintahdeverfteemnatlley not 2a5n0inanlg/k(gNo.thaFt53w4a54s) satcrreaitfeidcedat obnrokDeany b2acdku.e tAolsaon, apthpearent tainsismuaelss cinolitheecteidnitfiraolm ttehset wterraensppolratcaetdioonn) drpyrioirceto(dbueriinngg stored in a freezer. 02884 Tohuetsceomedevoifattihoensstuadrye. not considered to have had an adverse effect on the 000033 @ WIiSTONSIN ) [OSIRIS SH Page 29 of 45 4 CORNING Comma Sponsor: 3M ToxicoloSgty. SPearuvli,ceMiMnendeiscoatla Department PROTOCOL TP3016.A8 StudyTitle: Single-Dose DermTa-l60A53bsoirnptRiabobni/tTsoxicity Study of ) Date: Decenber 30, 1994 PerforningLaboratory: Ha33z0l1etoKninsWniasnconBsoiunl,evaIrndc. Madison, Wisconsin 53704 LabPorojreca tIdetntioficratiyon: HHI 6329-137 03885 J | 000035 EPXhPoRnEeSS 6M0A8IL241DE4L4I7V,ERY 3301 KINSMAN mivn waoiFsaoxn wooun zurasiidss i ) J Page 30 of 45 ~ STUDY IDENTIFICATION TP3016.A8 Page 2 Single-Dose Dermal Absorption/Toxicity T-6053 in Rabbits Study of HMI No. Test Material Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date DErxapfetrimReenptoarlt DTaetremination Date 6329-137 T-6053 Departnent 3M Toxicology Service Medical 3M Center, Bldg. 220-2-02 P.0. Box 33220 St. Paul, MN 55133-3220 John L. Butenhoff, PhD 3M Toxicology Service Medical Department 3M Center, Bldg. 220-26-02 P.0. Box 33220 St. Paul, MN 55133-3220 (612) 733-1962 Steven M. Glaza Hazleton Wisconsin, Inc. P.0. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Hadison, WI 53704 HBuaizlldeitnogn NWoi.sco3nsin, Inc. 3802 Packers Avenue January 13, 1995 February 10, 1995 March 24, 1995 038s6 000036 Page 31 of 45 - ) TPaPg3e0136.48 1. SStiundgyle-Dose Dermal Absorption/Toxicity Study in Rabbits 2. pTuorpaossseess frritancy otfhea styessttenmiactearbisaolrpwthieonn aanpdpliteodxictoitytheandskirnelaoftivreabsiksiin 3. Re[LghaibusolraastttoourrdyyycwPoirmlaplcltiibaceneccRoenedguucltaedtiionnsa/cScotradnadnacerdwsi/tGhuiitnhdeees.lfollowing Good b[d1 C2o1ncdruectseas(afonN)onregulated Study [[1 14400 CCFFRR 719620 ((EEPPAA--TFSICFAR)A) [[11 [1 N5o9(t8iN1fo)ih3cs0aatn(iFoNinon.alN)o3.85(03O1E3C"(DJ)a(pJaanpeasneeseMAFHFO)KN) dAWiTerIlefcaptrroeorc,AecdtutrheeRsegsutilunadtytihodinosse.sprontoIotncotulhneneacorepeisnsiianornicloyomfpdluitpahlenicceSaptwoeintshoanrytahpnerdeAvn`iistomuuadsly work. ) 4. QTtuhhaeelipQtruyoaAtloisctsoyulr,AasnsscutreuadnyceconUdnuitct,inanadccotrhdeanfcienalwitrhepHoarztlewtiloln Wbeisacudoinitsyed by (HW) Standard Operating Procedures (SOPs) and policies. 5. TestMaterial A. IT-d6e0n5t3ification 8. Ph(TyosibcealdoDceusmcernitpetdioinn the raw data) C. P TdheeteurSmpiaornnnasdtoSii rotnaassbtsiu(lmiiyentscylurdeisnpgonusnidbeirlitteystfocronpduirtiitoynsa)n.d stability 0. SRtooomragteemperature 03887 000037 Page 32 of 45 , TP3016.A E. ReserveSamples Page 4 mRaetseerrivaelssawmillel(sb)e otafkeenachforbattheihs/Tosttudoyf. test and control {IMeI itnesta fanrdeezceorntrsoe]t 1f0oryelaorssears per HAI StmOoaPt.emraiianTlhteairnSepsoaenrsvtoeermpsweairmlapltluersbee wocilfolnt-a2bc0etesdtaosryeeetdrorat regulatoryfoGrooddispLoasbiotriaotnoryinPraaccctoircdeasn.ce with the sppropesatn F. RAneytenutniuosend completion otfestthematienr-i1ailfewiplhlasebeofretthuerne`sdtutdoy. the Sponsor after 6. SaAsferteyqPurireecdaubtyioHnIs SOPs and policies 6. ControlMaterial A. DIidsetnitlilfeidcawtaitoenr ) B. PhClyesairc,alcDoelosrclreispstiloinquid C. cP Tohnesu ipduerar rinetddySittaonadbt biesltiy aatdbyeiqluiattyeofforthitshempaunrupfoascetsureodf mtahtiesrisatludyf.s D. StRooormagteeCmopenrdaittuiroens E. ReSeseerSveecStiaomnpl5e.s E. Reserve Samples F. ARanneydtewrnietlmilaoinnnoitngbecondtirsoclardmeadteraifatlermafyssbueancuesedofftohreotfihnearl treesptoirntg. 6. SaAsferteqyuPirreecdaubtyioHHnIs SOPs and policies 7. ExperimentalDesian A. Animals (1) sRpaebbciites ) (2) Strain/Source Hra: (NZW)SPF/HRP, Inc. 03888 00003% Page 33 of 45 .) (3) AaIgdneuTaitttiation (9) M2e.i0atht03a.t0 Ikngitiation (5) N1u2mbmearleasndanSdex12 females (6) IInddeinvtiidfuiaclatniuonmbered ear tag TPaPg3e0165.48 (7) Husbandry (a) HI(ohnuedasivviyindgugsalulgye,) in screen-botton stainless steel cages (b) FAt(hoPemHoIedmaasFneuuerdfesad,ctaumIroneucr.n)t.foorfTnhLueatbrfoiortoaidtoonr1a3yl RrcoaoubmtbpiiontneelDnyiteetanaHarFtyys#e5d32e5y ) environmental contaminants. ater 4phfaoadrrtdeTnrsieebslaisert,ecutneaadnnfdarSelolypmezeneacedninftbaisyue,tdHohmImeaiatcvifrycoorbmsietyaoslttaaeglni.cdailsSsaocmtopnrlteeudsnisooflmiticrhee, organophosphates, and chlorinatetadlsh,ydrocarbons. (@) CtE hoanttawmois unladntisnterferecownittahmintahnistsstinudtyhe food or water ) e) EsBentvrirtooenmmeaeinntntaincoanttermoperfaotrurteheofani1m9alC relative humidity of 50% 420%, `and frooo33w"i.ll a be Tight/12-hour dark cycle. a 15-hour. 0 f) Afccelilmatoionn s (8) SaDBdeaielseqeenucdatttioeeon,nhneunoasflbtesThrbevstiaoonfudAsenlxbiytomrdaapylosowarneiihmseahaltlsawciclolrdibengputrochWaWIsedSOPs.thoArn ) dasantyirsmaatliosffiewsditluldbyodbyeinwipetliiaagctheitdonri.anntodomsitzuadttyihognriospurppaslgarcuasenidnguointahtienst.nineThe Goa|889 000039 Page 34of45 ~ ) TP3016..28 Page 6 (8) HJtihseutoarnsiicnaatlllyio,fffc"othrhoeiiScNpeceewcbiaeZecesaatSuTseaeinldeocoWfthiitnothnee allabrgienoamroaubnbtitofhas been background information on this species. B. DoAsdeministration (1) Test Groups --Srow TestMaterial _D(omseo/Lkevae)l MNalues mofbAnfeiemmparilesr 21 (Control) DisTt-i6ll0e5d3water 3 T-6083 1o0x 125 33 3 33 3 4 T-6053 250 3 3 *2,* TToo bbee aadduniinniisstteerreedd aatt aa ddoossee vvoolluunnee ooff 2..010 nlL/kkyg ) (2) ePOianfcrhntehrceeaebsdpbsaiaytraoyBwefirf(lEotlxroaepoboebttsteuacsirilteniApompuarentendebraliefamrliesehaeopdfplihsackiairtn.)i,on,thetheflabnakcsk oafnd, sIinu1sr]pfeacccteoendsarteifaot.ruteiTnhtaeeprpfrteorrxeiaintmgmaetneltleysisoi2nt0se%,s ofi(rirntithteaactttiootsTnakh,lien)obsrohwdadiyvleelfdecbsteesca atnhiamtalwsouwlidllprebeclucdleipptehde uasseneoefdeadnythorfoutghheouatnintahles.StudTyh.e (3) D"oAArilsdeTlsem1piaebncneitimiadvsleetssritgewansittalltieoodrrneaccsoenitDvraeoyal 1.m"saitneTgrhlieealda.odsmeiTnhfioesrtdraaeyatcihoonf torfeatthmeent Gatrouapdo2sewilvlolubneediolfut0e1d each animal in Groups 3 mwli/tkhg.distThiellerdespweactteriveanddoaasnphipm}aFlairedjn and 4 will beapplied undi)uted. wMteh1ieghdatroesaejsusotfinebxeGpfrooosruueprsead1ni-ni4nawiislttlrhiantbieoannbdasauenddniwfoionlrlnthaebelaansyipemrrae.ald'sTohnbeg.oedy a1r0e-acmoxf a10p-pclnicgaatuizoen b(aGnrdoaugpes s1-e4c)urweidllwitbhe 211 edges and overwrapped with Saran pcaopveerredtapweitharound btea'pecoltloarperdovdiudreinagn tohcecl2u4s-ihvoeurdreaspspilnigc.aNrtaipoTnheapnerdraiboEbd1i.atsstowpi]la}st (4) RTehaesodnernfaolr humans. Rroouuttee oifs Aa dpmoitnsetntriaaltiornoute of exposure in \ J 03390 000030 Page 35 of 45 ) TP3016.AB Page 7 C. ObservoaftAniimoalns (5) Reomf To esv tMata eril al Aapppprloixciatmiaotnelythe24bahnoduargsesafatnedr ctoelsltarosr wciolnltroble rmaetseervieadl and the residual test material an appropriate solvent, if will be removed necessary. using water or approximately 1, 2.5, and 4 hours after test patorias (1) ClinicalObservations For clinical signs before test or control material administration and for clinical signs and mortality at administration (Day 1) signs, and twice daily and daily (a.m. and thereafter forclinical p.m.) for mortality at least 28 days. Observations may be extended when for directed by the study director. (2) Reoa fDed rmali Irrn itatg ion Before dermal test or control material administration the initial irritation reading will be made and recorded as the Day 1 reading (Attachment 1). Additional dermal ) irritation readings will be made approximately 30 after bandage removal (Day 2) and on Study Days 4 minutes and 8. fIonrdiveiadcuhalanidmearlm.al irritation records will be maintained . 3) Body Weights a(Fpwophrelnircaanstudirovominivzaa(ltDiaoyenx,c1e),ebdesofnor1 Deadaytye)s29t, o`rancdonattroulnmsachteedruilaeld death (4) sample Collections (a) Frequency ) Before initiation (Day 1), approximately 24 hours post-dose (Day 2), Days 4, 8, 15, 22, and at experimental termination (Day 29) b) Num ofbAne imar ls AN 03891 < 000011 Page 36 of 45 ) PTaPg3e0186.A8 (c) 2M Bf,lroo4moe,dto8hs,teafmpCm1l5aohe,rlsg.liaeonn(cadatlpid2po2er.naorxiAvmpeapitrneolxoyf4emiLt)herwilelarboencboalylsect1.ed b{eacotbuatlainveodlumferontothbee animal sacrificed in pdoocsutmeernitoiermdavteeinnlaytch2ae0varnlaowfodfeatcbaci)hsodwill a moribund condition or csseaanmctprrliieffsiucgweeiddl,latabnetdhestthoteriendesepaotafrarntoeeocmrsoeptrseuymnpe(raDanadtyurc2ee9l).jaunldaTrhtehen f<fr2ra0acctCtiioonn1ss0Cws.itlolrTebhdee issneeanptarfafrtreeoezdzeenrseorsnuemtdratnyod miacceienltltoauilnathre Sponsor after experimental termination. Samples will be shipped to: 3JMameEsED. EJPoChn.son 9B3l5dg.Bui2s-h3Av-e0n9ue St. Paul, HN 55106 ) tJthaeenleespshamoD.pnleeJsoa.thns(o6n12)or7a7l8t-5e2r9n4atperiwoilrl tobethneotisfhiipemdenbty of D. Pathology (1) UngmArosnorycsihsbaeundnniudnelaclercodoSnpddsyaiyictnrigeioxfnadimucirwinielansltgainobtdnehDeaesnusadbttjuheadslcylteodarbntsooarcmaranilfiaitbcibeerdseviiwnialtaledb tarhneeecsomtradhreegdti.inzaeldAnewiiamtarhlsvesioinnd)i,uambmloperedintbovubinaadrbctihotenadlvietni(aovniacawviialn,ljeacbntedion e in ADe.fxtseaPrnagtuhnioenlcaortgoyep,dsy.,(3)tThiesSsauamenpsil,mealCasosllwdeielcsltcirobineb,eddiwsiiclnlarsdbeeecdt.icoonllected. (2) ScAhtetdeurlmeidnaStaicornifoifctehe surviving aninals will exbeperainmeesntthaeltizpehdasweit(hDaysod29i)u,n bbplerenedtovbviaitrabeitdthaelgrvoe(snvsaiaJcaeivcnarj,oepcsteyixosnaenxgaiumniinntaahtteieodmn,a.rgainndTahlesuabcnjaiermcatvleesdin)bt,blan be necropsied be recorded. in random order and a11 abnormal ities wi 03892 000042 Page 37 of 45 ) P3a0g1e69.48 (3) wpTsraaosimchpedelddeuerwCheio.tlhleT1chuEtekiSeoEwwnhaoralndetTacipovneatrtr,eerbimlapetr,eirotraan)tosiphpecapatciroennchwi be panLlne-aicccmoneapldsssie,icidntiaaoninndfreboeaofectzhhthgekrioddunepremywasllfr3opmpbleitcchoaeltilfeoicnrtsaetapdpimrroeeaxlniedameriwrsamedes''eddtc)sommniatnetxy d"i2s0caCrd$e1d0.C. Afteerr nseectrotposym,aintthaeinan3imtee)speerrcetsirsneoo) asTKhfihitdepneperteyisdes)xsptuwoeeirslitlmh(eeinbvtepeaerlrs,seotnnetbrimlTfeiir,nsoatztedeidenornmai.annlSdreTaychp.etpiliosicnacema7ptt.sioceo.rnt(h1ies)iytsiee,ansor oSfponthseorsaimsplreess.ponsible for the retention and d1.5i(cc)r-itinT,he E. StNaotsitsattiicstailcAanlalaynsaleysses are required. 8. RA'eFpionrat] report. including those ftems 11sted below will be submitted. ) DPDreeosscccerrdiiupprtteiisoonn ooff tthhee tteesstt asnydstecomntrol materials DTDTaeaabstbuceulrslaiatptoitifoiononenxoopffoefrmmiemoaaernnntytaalbtloiodxtyiyicnwdieaetitfigafahettcibstoysn/bsdyeaoxnrdsneaaxnt]gearndm{deirmrndsiaottcaijst.oonoeeln Gross pathology findings/aross pathalagy report vel 9. aOLfcraociccegiapiltntiiaatolnancteedoaftoaafuR,datiwhtoeirDantgFcaio,ntpahileeRsecrsoettrpuhdodesryr.te.odfua,rniWdhnweigFnlilnitats)h.BeepRrseovpgaorireTtsasblesndatbHrumIireto raelltaionreidginianl thpeapaerrchdiatvae,s oifncHlHuIdinagccothrodsien.g itFtoienmaH]H1I5r5seepdpo.rtbeloncoamrpiletLecd, DPIornso-etToicfpoerlepreaacrnoadrtdfpsornotorceoclordasnendnents pulDBobossdeyerwaaedtinigiohnntissstration . ) SShanimabptploiennigccaollrleecpcoatrtdhosonlogryecorredcsords Study correspondence 0y 3893 Final report" (original signed copy) 000013 Page 38 of 45 ) TP3016.A8 Page 10 The not following supporting be archived with the records will study data. be retained at HHI but will Animal receipt/acclimation records Water analysis records RAneifmrailgerraotoomr teanmdperfarteuerzeer antdempheurmaitduirtey records records Instrument calibration and maintenance records ) 03894 000031 Page 39 of 45 ) 1Pa7g3e01116A. PROTOCOL APPROVAL I 3SoMpnoTnoTsxoircsoBliotRgeeynphroSefesfre,vnitca0 etiMve--edical Department 1-4 oe gp ASScttuuetdveyenTDoKi.xrieccGotalozoragy 5 Hazleton Wisconsin, Inc. ) 4s resentative : HQauzalleittoynAsWissucroannscien,UniItnc. (6329-137. protdskz) Bite ee bite 2/ap Jot C03895 000045 Page 40 of 45 ) TPaPg3e01612.48 Attachment 1 Scoring Scale for Acute Dermal Reactions Ervthens 01-- oslnieght 32 -- SHeovdeerreate Edema 0172- MSNolodinegerhatte(ba(rraeilsyedpearcpepprtoixbilmeatetloywe1llm)defined by definite raising) 3 - Severe (rafsed more than 1 nm) Atonta ) 201 --~ SNToingeht (slight Hoderata (slow rfemtpuarinrmteontnoofrael]asticity) 3 - Marked (no elasticity) Desquanation 01 2- MSNolodinegerhatte(sl(isgchatlesscaalnidngf)lakes) 3 - Marked (pronounced flaking ith denuded areas) Corfaceousness 01 2- MNSooldnieegrhtate(de(clreeaatsheeryintepxltiuarbei)lity) 3 - Marked (tough and brittle) Eissuring H= G Slight (definite 2 hoadraed TERNS cracnks yin epidermis) ) 3 - Marked (cracks with bleeding) 03896 000016 @pnziemay yo BIRT NEN 8 Page 41 of 45" +CORNING Carry PROTOCOL TP3016.AB Single-Dose DermTa-l6A05b3soirnptRiaobnb/iTtosxicity Study of Sponsor 3M Toxicology Service Medical Department 3M Center, Bldg. P.0. Box 33220 220-2-02 St. Paul, MN 55133-3220 Soonsor's Representative John L. Butenhoff, php HWI 6329-137 Contractor Hazleton Wisconsin, Inc M3a3d0i1soKni,nsmWaIn 5B3o7u0l4evard StudyDirector Steven M. Glaza ) Anendnent No. 1 This anendnent. modifies the following portions of theprotocol: 1. Page 6, 7. Expe : B. Dose Adni don; oups. Effective January 13, 1995 The test material needed to utilize mixture could a dose volume not be of .01 prepared mL/kg. at the concentration asterisked change: footnote (**) in this section with Modify thedouble the following underlined ? faaesor boerinental Desion: b. Dose Administration: (3 Dose ** To be administered at a dose volume of 210 mL/kg Administration, The Concentration needed t ttoestutimlaitzereiaal domsiextuvroelumceoulodf not be prepared at the hird sentence in this section with the following .0] nl/kg. Modify under] ined change: the The dose for each water and applied animal in at a dose vGorlouumpe 2owfil1l0 bmeL/dkigl.uted With distilled ) 03897 EPXhPoRnEeSS 6M08A241D.0e4v1)en 3301 ximsMan avo 000047 maoiFsoexn so`utu sai rae; Ak Page 42 of 45 .) Amendment No. 1 HHI 6329-137 Page 2 Effective January 17, 1995 3. 6.7. sDAeadcymoinn2didsuterattoionan. t: iB, finistration; iAnnjiumrayl (Naop.parFe5n3t454bro(kGernoupbac4k)f.emaAled)d wjausg s Fsoalclroiwfiincged on paragraph to this section: as the rD(euApenliamtacolenteNhnoe.t saFan5ci3rm4ia5fl4i)cweibleolcnaubDesaeytro2efaotafendonieantjuGtrrhyoeup(saa4pmpefaerdemonastleebJroovkee]n same manner jboackc)h,e a bDoodsyeTvwaetiigohntss aas(ncdlfionsriacmtaphllee oobctsohelerlrveacattniiioomnnassl,]s 3ninals {unscheduled rainendadtithnhege spotauftdyhd.oelromagTlyhoifrriirteation, tahned ostahmeprleacnoilmlaelcstiionn)tshawecirlsiltfuidcbyee.scoanndducdteaetdhsi,n tshcehesdaumleedmasnacnreirfiocce,For Effective January 24, 1995 ) svooeFofcltubpimihoelensesrwceioqllulleesctbeteodfinwtcihlleuldeSdbpoenwsidotorhc,umthetenhteesdawmepiilgnehttshsehioprfmaewtnitds.astuae.HsodciTofhlyelsesschteewfdeoiloaplnedoswtinhneegdvolume of the protocol to include these additions. 4 HsoedctiifoyntwhieEthsxecthoendfoslelnotweinncge uinndet5rhDle.inTPeidarsttadhdainotdilosn(eosc3:og)nSdyap:maprlaegreCpohlslecotFioots tdIehhreenaFw]ihroslateppmlTaivlceeart,iaonndbilsfeei,mtaelaenfrnoaempcprraololpxsiimaeanditmeailn1s-,ecancahnxdg1rb-oocutnph sweKiciltdlnieobynys oFfratnhe co-il2m0lmeeCdcit+ae1td0elCwy.epilcahceedd(vinolaumfereoenzleyr dseettertmoinmeadifntoarinbialet)e.mpearnadture of sThheippseadmpltoesthaendpetrhseoinr TciosrtreedspoinndiSnegctiwoenigh7ts . (o4r)v(c o0l)uses will be 5. Page 9.8. Report. Add the following to this section: Individual animal tissue weights and bile volunes 03898 ) 000045 ) Anenduent No. 1 Page 43 of 45" PROTOCOL. AHENDHENT APPROVAL HPHaIge63329-137 [Zi] Loitotilf 3JSMophoTnnosxLoi.rcsoBluotRgeeynphroSefes?re,vnitcaPetRiMvOeedical Department 2/5 Date SSttuedvyenDiW.recGtaozra AHaczulteetoTnoxWiicsocloongsyin, Inc. ) Jue, the ReQpuraelsietnytaAtsisvuerdnce Unit Hazleton Wisconsin, Tnc. (6329-An1d.3ds7k2.) ate 2-695 Date 2.7.95 az J cee 03899 000019 Page 44 of 45 - @ WISCONSIN ) USBIBL ian + CORNING Conny PROTOCOL TP3016.A8 Single-Dose ODFermTa6l05A3bsoinr'pRtaibobni,Ttsoxicity Study WH 6329-137 Sponsor 3MMeTdoixciaclolDoegpyarStemrevnitce 73.M0.CenBtoexr,432B2ld0g. 220-26-02 st. paul, WN 55133-3220 Contractor M3Ha3ad0zi1lseoKtnio,nnsnWWIainscoB5n3os7ui0ln4e,varIdnc. John L. Butenhof, php Steven M. Glaza Anendnent No. 2 ) This anendnent modifies the following portion of the protocol: Effective January 20, 1995 1. PL3m3aaa0big1onert2aaKtiionnrseSymdt,aunda3ty8B0ot2uLholecePavamtaicaorkinden.,rsfaiTcnishtleeiatrdyepoolffaceaHtmaezHnltasztloaenntiomNnaylacNoiwnsasesionnt,srienai'trseed,Acauntde Studies JFoelcleoiwviendg:and acclimated iAnvetnhuies,fasciinlciety.the rReeppllaacceenetnhtismsienctsionLossith the Stu(dInyitLioaclatTieosnt) Stu(dRyeplLaocceamteinotn Animal) 3HB8au0zi2lledtiPonagnckeWNroi.sscAo3vnesinnu,e Inc. Madison, WI 53704 M3Ha3ad0zi1lseoKtnio,nnsmWWaIinsco5Bn3os7ui0ln4e,varIdnc. 03300 ) 000050 PCrEoRnEeS sWoAyNzai ricees r own man wen waorFsasxn wun at treLas 3 Amendment No. 2 ) Page 45 of 45 ~ HWI 6329-137 Page 2 PROTOCOL AMENDMENT APPROVAL Glow 2. Riiltf ry John L. Butenhoff, PhD Sponsor's Representative 3M Toxicology Service Medical Department Date Deo 4 Steven M. Glaza ) ASctuutdey TDoixrieccotloorgy Hazleton Wisconsin, Inc. Snax Date fe su. Representat je HQauzalleittoynAsWsisdcroannscien,UniItnc. (6329-Am12.3ds7k3.) Arras Date 03901 J 000051 9.1.2 Analytical protocol AMDT-022195.1 03902 000052 3M Environmental Laboratory Protocol - Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T-6053 in Rabbits In-Vivo Study Reference Number: HWI#6329-137 Study Number: Test Substance: AMDT-022195.1 FC-99 (T-6053) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facility: 3M 935 Environmental Bush Avenue Technology and Services St. Paul, MN 55106 Proposed Proposed CInoimtpilateitoinoDnatDea:teJ:ulAyu2g5u,s1t92955, 1995 Method Numbers AMDT-M-1-0, and Revisions: Thermal Extraction of Fluoride by Meansof a Modified AMDT-M-2-0, Dohrmann DX2000 Organic Halide Fluoride Measurement by Means of Analyzer-Liver an Orion EA940 Expandable AMDT-M-4-0, Ton Analyzer Extraction of Fluorochemicals from Rabbit Liver AMDT-M-5-0, AElneacltyrsoissproafyRMababsist Liver Extract Spectrometry for Fluorochemicals Using AMDT-M-8-0, Analysis with lon SoefleFcltuiovreidEeleUcstirnogdethe Skalar Segmented Flow Analyzer Author: James D. Johnson Approved By: baer ames on Study Director ls Date eo3 203 Pl2. Bil fff ai n John Butenhoff, PhD Date Sponsor Representative 000033 cdTohenirtsmaaislntliuyndgyabcissoodmrebpseodiu.gnndeTsdhe(tobaipnoratolryvatinidscefaolirnamfsaoptreimcoatntopifrtoonhdiaussctstsot)uwdihnyetitshheetortidFsesCtu-ee9ra9mnid(nTeS-ef6lr0uu5om3r)ionfies. rabbits used to amtavkaeriaonusastsiemsessmpeonstt odfostehedeexrtmeanlt aopfpdleicramtailonabosforFpCt-i9o9n. The data of FC-99. are to be 2.1 Tes2t.,1.C1onAtnraolly,tiacnald Reference Reference Substances Substance: and Matrices FC-95, lot 161 equivalent. or 171. They are 2.1.2 2.1.3 AAnnaallyyttiiccaall RCeofnetrreonlcSeuMbasttarnicxe::BNovoinnee liver and bovine serum 2.1.4 Analytical Control Matrix: Bovine liver and bovine serum l2i.v2erS),ouSricgemaofCMhaetmeirciaallsC:o3mMpaInCyP/(2P.C1.P2,Di2v.i1.s4iosner(2u.m1).1), grocery store (2.1.2, 2.1.4 2a2.n53d0 Nm6ugcm/oknbtgerordloosafeniTgmerasoltus.panwOdansCeoranenptliramocaleldSwawamisptlhreeaspn:liamTcaielsdsFou5ne3s5d0aa4ny)d.2fTl(uiaisndsisumeafslroaFmn5d3148f5ltu4eisditsnaitnnhicemlauldse liver, these tkiisdsnueeys,wsilelrubme,atcetlhleuldairsfcrraecttiioonn,ofdetrhemaSltuadppylDiicraetcitoonr.site and bile. Analysis of t2h.e4 HIdWeIntiafniicmaatlioindeonftiTfeisctataionndnCuomnbterrolwhSiacmhplceonss:isTthsoefsaamlpeltetesraarnedifdievnetidfiigeidtusing 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. 2.7 -20:+ Sto1r0agCe(C2o.1n2d,it2i.o1.n4s).foTreTsetsatnMdaCtoenrtiraolls:saRmopolems AMDT-S-10-0. tweimlplebreatruerceei(v2e.1d.1a)c,cording to 03304 000051, G2.L8PDiRsepgousliattiioonn offorSptehecitmiemnesp:erBiioodlorgeiqcuailretdisfsouresstaunddiefsluliodnsgweirllthbaen r2e8tdaaiynesd.per 2.9 Safety attire. Use Precautions: caution when Refer to appropriate MSDS. Wear appropriate handling knives for cutting the samples. laboratory `The tissues analysis for from animals dosed as described (HWI#6329-137), are available for fluorine compounds. At the discretionof the Study Director, a series of (aSnaeleytMiectahlotdess-ts-nceaxntbSeecpteirofno)rmisedt.he Tahpperospcrrieaetneinagnafloyrsifslutooripdreesiennltidveefrivniiaticvoemdbautsatifoorn oflfutohreinsetuindythien ltihveero.piWnhioennosfutffhieciSetnutddyatDairheacstobr,eeannacloylsliesctweidlltocemaesee.t the objectives 4.0 EXPERIMENTAL - Methods 4.1 Liver and Serum screening methods:(attached) 4.1.1 AMDT-M-1-0, Thermal Extraction of Fluoride Dohrmann DX2000 Organic HalideAnalyzer-Liver by Means ofa Modified 4E.x1p.a2ndAaMbDlTe-IMo-n2A-n0al,yFzleuroride Measurement by Means of an Orion EA940 4.1.3 AMDT-M-4-0, ExtractionofFluorochemicals from Rabbit Liver 4U.s1i.n4gAEMleDcTt-roMs-p5ra-y0M,aAsnsalSypseicstroofmReatbrbyit Liver Extract forFluorochemicals 4.1.5 AMDT-M-8-0, Analysisof Fluoride Analyzer with Ion Selective Electrode Using the Skalar Segmented Flow 5.1 Data Reporting: Data will be reported as a concentration (weight/weight) of tfilsusoureidoer pfelruitdi.ssSuteatoirstfilcusidu,seodr,aast FtCh-e 9d5is(cerleetcitornoospfrtahye mSatsusdyspDeicrterctoomre,trmya)ypeirnculnuidteof 603905 000055 3 regression deviations analysis of serum of concentrations concentrations with time and for different dose groups. If averages and standard necessary, simple statistical tests difference. such as Student's t test may be applied to determine.statistical 6.1 Raw approved Data and Records: Raw data, final report, and electronic data approved protocol, will be maintained appropriate specimens, in the AMDT archives. 7.1 AMDT--10-0, Sample TrackingSystem 8.1 AMDT-M-1-0, Dohrmann DX2000 Thermal Organic Extraction of Fluoride Halide Analyzer-Liver by Means ofa Modified E8.x2paAnMdDaTbl-eM-Io2n-A0n,alFylzuoerride Measurement by Means of an Orion EA940 8.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver 8E.l4ecAtrMoDsTpr-aMy-M5a-s0s,SApneacltyrsoimsetorfyRabbit Liver Extract for Fluorochemicals Using 8.5 AMDT-M-8-0, Analysisof with Ton Selective Electrode Fluoride Using the Skalar Segmented FlowAnalyzer 03906 000056 4 3M Environmental Laboratory Method Thermal Extraction ofOFrlguaonriicdeHablyidMeeaAnnasloyzfearM-oLdiivfeired Dohrmann DX2000 Method Identification Number: AMDT-M.1 Revision Number: 0 Adoption Date: /o- f-1 Revision Date: None _-- Author: Rich Youngblom Approved by: - foup Leads a /o/3/8< `Date Luh2 rs Quality Assurance Software: MS Word 5.1a f0o--yfg Date Affected Documents: AMDTE-xMp-an2daFbllueorIiodneAMneaalsyuzerrement by Meansofan Orion EA940 Organic Halide Analyzer 03907 AMDT-EP-3 Routine Maintenance ofaModified Dohrmann DX2000 1000057 - 0 SCOPE. APPLICABLE COMPOUNDS, AND MATRICES wf1ilutohSraicdnoepiefo:rnoTsmehlivesacrtmiievoetushelomedacttrirsiodcfeeo.sr.thTehoepefrlautoiroindeoisfatyDpiocharllmyacnonllDecXt2ed00in0TwIhSeAnBitsiosluutsieodn tfoareaxntarlayctsis 1.2 1.3 MAaptprliicceasb:leBiCoolomgpiocaulntdisss:uesF,lupaorrtoicchuelamrilcyallisveorr. other fluorinated compounds. 2.0 KEYWORDS 4DX2F0u0o0r,idfel,uofrloucohreirnmei,ceaxltsr.action, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, 3.0 PRECAUTIONS 3.1 3.2 GGllaasssswwaarree iasnfdraegxihlea,usbtrogkaesnesglcaasns bmeaeyxtcraeumseeliynjhuorti.es. 333:::543 PSPoortleevsnsetunirtailzbbeaidsoeghdaaszseaasrm,dpsplredosupemertaocytohfmelpabrsiheo,slsmoegadiycganalesmeahdtartnoidcalelsi.lngowUprstaectahipctpereosodprmerrqiyuaitdreeodpw.enrsboenfaolrpersottaercttiivneg.run. equipment, 4.0 SUPPLIES AND MATERIALS 4.1 4.2 CCoommpprreesssseedd Oxygen, Helium, HHiygdhroPcurairtbyoGnrfardeee,, regulated regulated to to 30 45 PSL. PSI. 444:834 QOQuruaiarortntzz9gg4ll0aa9sss9s9csoaTmmobtpaullsetTbiooonanitctwSuibtterh,enRTgeetlfhilaAondcnjeTMusGttluambseisnngt,GB-Du9of4hf0er5rm-(a0Tn1In2S8oAr9B0e-qI0uI)i9v7oarloerenqetuqiuviavlaelnetn,t, 44..67 MSaimlpllie-cQolwlaetcetiron vials, HDPE. 44..89 PAocltyisvtaytreednCehapirpceotatels,.E. Merck 2005 or equivalent, 44..1110 HMiasmcielltloanneSoyursinlgaeboorrateoqruyivgallaesnts.ware $:1 5.2 IRoBsMemcoomupnattiDbolher3m8a6nnorD4X8260c0o0mpOurtgera.nic Halide Analyzer, modified for fluoride extraction. 5:3 5.4 DExXc2e0l 0S0presaofdtswhaereet,, version version 15..000,ormgordeiaftieerd for fluoride extraction. 6.0 INTERFERENCES `6:m1ay Svaamrpylferosimzemaitsrliixmtiotemdattoriaxp.proximately 150 mg, depending on sample moisture content. This 03908 2 000055 1.0 SAMPLE HANDLING s7a.1mpSlae.mplUesseafroerncoetpstoorbeprhoabnedlteodtrwaintshfebrartiesshuaensd.s. Fluoride may leach from the skin to the c`7ml.ee2tahSnaansmcopalll,peoesrlofmaenltdihvcaeunrtoatlri-enwgacbutotearrfdrs.oolmuTtfhireoonzceuaftnttleiirnvgeearbcaohanrdldipvalernadciessdccuaitln.paeltasrheoduladndbelacbleelaendedwewiigthh bwoaatte.r,Usea 8.0 CALIBRATION AND STANDARDIZATION 8.1 Preparation ofCalibration Standards 8R.e1f.e1rTthoeprsottaoncdoalrdfsorrtehqautiprreodjefcotr.each project will need tobe appropriate for that individualproject. 8$1.5110.32mgFToy)priarcnaadblwbleyiil5gi0hv-e5rit0si0tnupadpilcmasbFeulCse-edb9ea5enifdnltmiavereterhdaawsneoitlhgehstmbaaotnardti.axr.dsCaurte auspeide.ce of fozenbeefliver (100 - 8.2 Calibration - Overview `The normal calibration isthe fluoride curve curve is required the procedure listed below (AMDT-M-2). is used. However, ifan optional spiked liver 8a.n2d.1coAmbcuasltiibnrgattiohnecmuursvienfgortthehesaDmXe2m0e0t0hoisd. gseannedramtaetdribxy tsyppiekaisngthse.amspalmepslewsittho kbenotewsntsedt.andards s8p.i2k.2edT.ypically, three replicatesofeach standard and five concentrationsofstandards will be R8.e2c.o3veSrteadndFar(dugc)uornvethwielyl-abxeisp.lotGteenderaastMeaasrsegSrpeiskseidonFc(uurgv)e oanntdhecxa-lcauxliatse atnhde and the 12 value. Standard Mass equation for the line. 8:24 Mass Spiked F (ug) = (Amount spiked in mL) x (Cone. of standard in ppm) x (0.6004) 8.2.5 Standard *FC-95 is 60.04%F Mass Recovered F (tuhge)re=fo(rTeI0S.A6B00v4oilsutmhee ifnacmtolr)usxe(dOtroiocnonrveeardtinFgC-in95pptmo )F 8.3 Calibration - Procedure 83.1 Start Up b8.e3.u1s.e1diRfutnhe2porrevmiooruse sCalmepalneCsyccolnetsaiwnheednhisgtahrtcionngceinntsrtartuimoennstoeffalcuhordiayd.e. More clean cyclesmay 8.3.2 Blanks 83.2.1 8.3.2.2 Prepare sample using the same methods and typeofmatrix For rabbit studies, usebeefliveras the matrix. Prepa as the test sample. (100 - 150 mg) for blanks. re at least 3 samplesofbeef liver 83.2.3 analyze PsuatmpslaempalcceoirndiDnoghtrommaentnhobodatA.MDCTo-mMb.u-st2 efaocrhthseaimopnlesealsecdteisvcereilbeecdtriondseeacntailoynsi9s..0 and GO3909 300005Y Jb8ue.df3.og2re.em4eprnFotocroerfeadbtihbneigtopwseitrutdahiteotsrh,ethctaehlerimbereaadttieinrognr.iesaBsdtuianmbglesfoawrmiaptlhberlseasnupnketicsltatmohpilsheilsitsmohirotiucilasldrrebeaeacdh0ie.nd0g,3sop(rppurmnetvoiilroilunsowtaheger 3ho3u.r2s.)5. For non-rabbit studies, the blank readings should reach a predetermined ion concentration `before 83.2.6 proceeding It may be with the calibration. necessarytomix approximately 50 mgofcharcoal with the sample to aid combustion. 883.3..33.1StWaenidgahrdouCtuatrvleeast 15 matrix samples (5 standards with 3 replicates each) intaredand 8sl3ta.ub3dely.e2ddatwPae.liagcShetobwroeeaittgsh.heemFdaotbrreirexafbslbiaivtmeprsltsueadsimeopsnl,dewrieynigDichoeh1or0rm0ia-cn1e5np0asmcakgmspbletoeefkbeloaietvp.etrhseammpflreosz.enRuenctiolrdwewede.ights in 8333 Start with the lowest standard concentration. Using a Hamilton syringe, ejecta fixed oqruainnttihteyboefetfhleivsetra.ndard on or in the matrix. For rabbit studies,use 4 ul ofstandard and eject iton 8.3.3.4 may be Atleas3t replicates used at the discretio should nofthe be usedfor analyst, the lowest standard concentration; more replicates r88e.3p3.l.3i3.c.56ateRC,uornmubnaulalsnt1o5tthhseetrasnsadaammrppdlsle.eIafosfdoetnshecatrriesbptelaidncdaairntdes.iesctIsiniodgninci9af.it3ceaainnntlddyaatdnaiaftlfhyearzteenttahcefcnrooermwditrnhegeplotiotchaAetMre DtrTewpo-lMac-e2s .the l3oe.la3ds3tr.e07p.l9i5Wc,ahtceeonnasnaudlllttshwtaiattnthdhaesrudnpseerhwvairvseeoplrbi.ecaetnerwuinl,l cbaelcuusleadtetothceal1c2u.lrat2emtuhestrebgreeastsiloenasctu0r.v9e5., [fit s not at 8.3.3.8 A new changed. New standard standard curve curve should be run when the combustion tube or `sample `may also be run at the discretion oftheanalyst. matrix is 8.4 Storage Conditions for Standards 8T8.y44p..i12caSlNtleoy,rwasgFteCan-rde9aq5rudissrteaamnreedsnattrosdrsfesodrhaostutlardnodboaermdptsreeamrppeaerdreaedtpuaetrnedleeiannsttplooannstctiehceasicmnrodeniwtvihtd.oupalbostttlaensd.ards used 9.0 PROCEDURES 99..11.1TyCpoimcbaulsOtpieornattuibnegtCemopnedriattiuornes:= 950C. 9.1.2 Oxygen and Helium flow = 50 ce/minute. 9.1.3 Vaporization/Drying time = 240 seconds. 9.1.4 Bake time = 300 seconds. 9.2 Start Up Procedure: 9.2.1 Ifthe program is not started, start the EOX program on the PC. 9.2.2 9:2.3 POuptetnhethfeumSaYcSe TmoEdMulSeEaTnUdPthweicneldlowin.the READY mode. 92.4 Close the SYSTEM SETUP window. 03910 a000060 PfoounndhinetnhethCeoEvLeLnCHhaEsCteKacmheend.the READY temperature, run the CLEAN BOAT program 9.2.6 See AMDT-EP-3 for detailsofthe Dohrmann software, ino9o.x3uplSadapempirpnsoloxeitmEhaxettSerlaAycMt$iP0omLnEgPrHoofAcceThdaCurrcHeo:aalndwiptlhactehetshamspalmeptloeaiindtchoemBbuOsAtTio.n.It ImFatyhsbesnGeocnees.scaroy taol 932 ClosebeSAmiMxPedLEin HwhAiTlCeHes,tablishing the basline and when generating the standard se. 23J.:a03b.em3leLddosdfam1pa:pl1peTrcoIopSlrAlieBact:teMiovinolllviuiaml-.eQTMToyfwpTaitIceaSrlAlmByix0stou.lru6et.Liontoor151:1mLTIaSrAeBu:seMdi.llFio-rQrTMabwbaittesrnamdiixetsurseeeto1a or 99in33c..h54es.RPluGanacsetehtseherEevOliXeala-ssSeodOtdLhuIartDitnShgepptryiorpogolrfyastmihsefmoCuuOsntdMBbinuUbtSbhleTeIRtOUhrNNouTmgeUhnBtuhE.e iTsIiSnAtDh,e TISAB at least 0.25 15o0r3.:t7h8.e ITWtIhuSenAdniBltuhtteoeEmdaOTkIXeSpA1r:B1ogwTraIasSmAuBiss:efdMinioilshcleoidl,-lQer,cemtohevesatmhpelceo,llaedcdtiaonnevqiulaflrovmoltuhmeeocfomMbuTstiGonPtusbee.er Lr$2i::et33hm.:.o98vReOciponatesntneoynttThhtIeeiSpespAnaedBmporlfeetmhaheaitncciohnmgabnuodsntrtiehoemnottvuubebe,eanwyitrhemMaiilnlin-gQaTMshwaftreormatnhde wbioapte. wiAtshhacKanIbMeWrIePmEovteod boboattttoomawiidtihnathsepaCtludelaaanoprCyopctlhele,riosircmviaalcartuudoemveircde.outA. dTt maoryfMbieolnleic-peQssarwyattoersemraapypabretiacdldesedojfofisre 99.33..1110 93.12 CRluonsethteheChLatEchA,N BOAT program. Samplei ready for analysis by ion selective electrode (AMDT-M-2), 9.4 Sample Calculations 994:.412 USsaemptlhee sMtaasnsdaRredccouvrevreetdoFcal(cuugl)at=e(tTheISsAamBpvloelvianlumeL.) (Orion readinginppm - intercept) (Slope) 10.0 VALIDATION ca1son0ona.ncl1tey.sniQtsrDuaaastililisytayrSttCeaodrntbtrUyopalnCalhyezcikngSaQmCplseasm:plOesn.ceTthhee QstCansdaamrpdlceusrvareeistoesbteabtlhieshseadm,eaacqhthdeayooopfest dreopnleicainteripslnoionctasnpeeicukenesldseassrsaym.thplefsirustsetdwtoorgepelniecraattees atrhee swittahnidnar2d0c%urvofet.heEascthancdoanrcdencturravtei.otnhsonosarcd mP2ar0tu2roirciPedrseecaNinosdtiedobinoffoaeknrsden2tA.c3r,cefuaernradecn3yc.e: cPSroeemcepiosmuienotdnhsao.ndddaecvceulroapcmyevnatriaensalwyhseisnaanndalsyazmipnlgesaanmapllyessiosfindifferent 10.3 Other Validation Parameters: NA 03911 5 0U00LL 11.0 DATA ANALYSIS 1L1 Calculations 14-1 For the standard curve, use regression analysis in Excel, version 5.0 or greater. 11.1.2 To calculate the fluoride contraction in the sample, see method AMDT-M-2. 11.2 Analyzing the Data a1r1e2.wi1thi2nm2u0s%t obfe cataclheaostth0e.r9a5nodrtghreeaotuetrl.ie"rOuistlgireerast"ermtahyanb2e0e0x%cloufdtehdeiafvtewroagoeftohfeththorseeetrwepoli.cates nleostsedthiannt5he0%FoinfatlhReepaovretraalgoeonfg twhiotshetthweor.easAonnyitswuacsh ocuotnlsiiedrsersehdoualndbouetlpioeir.nted out in the data and Leo arraceents None 00 0 00 13.0 REFERENCES 3143.91, rReovsiesimoonuBn,tDDeochermmbaenrn1D99X32)000 Organic Halide Analyzer Operator's Manual (Manual 915- 13.2 AMDT-M-2 Analyzer Fluoride Measurement by Means ofan Orion EA940 Expandable lon 13.3 AMDT-EP-3 Analyzer Routine Maintenance ofa Modified Dohrmann DX2000 Organic Halide 14.0 REVISIONS Revision Number Reafosr Choannge Revision Date -- 03912 000062 6 3M Environmental Laboratory Method Fluoride Measurement by MeaAnnsaolfyzaenr Orion EA940 Expandable Ton Method Identification Number: AMDT-M-2 Revision Number: 0 -_-- Author: Rich Youngblom Approved By: Adoption Date: /0- 445 Revision Date: None 00000 Grod] Leader 2L/5s Date Qual2 ity AsS suraL nceRAL LoDa-t#e F5 Software: MS Word 5.1a 'ADfofehcrtmeadnnDoDcXu2m0e0n0tsO:rgaAnMiDcTH-alMi-d1e AnTahleyrzmearl Extractionof Fluoride by Means ofa Modified 03913 J 000063 PE , APPLI( NI 1-1 Ton SAnCaOlyPzEer:. This method is for the calibration and operation of an Orion EA94E0xpandable 1.2 APPLICABLE COMPOUNDS: Fluoride. PLH3OAfP6.P0.LICABLE MATRICES: Liquid samples in an appropriate buffer solution. Preferred 20 KEYWORDS 2.1 Fluoride, fluorine, ion selective electrode 3.0 PRECAUTIONS 3.1 No hazards identified with this method. 4.0 SUPPLIES AND MATERIALS 444.:132 OOOrrriiiooonnn 9M94o40d09e90l979901To00t00a0pl1pTeomlneifccltuSrotordrieednefgistlthiannAgddasjroudlsutotmrieoennqtu(iABvguaCflfelen)tro,rIe(qTuiIvSaAlBenItI.) or equivalent 44..54 MMaiglnletii-cQswtiartbearrso.r equivalent, 44..76 SLaambpltiesscuoelsl.ection vials. 4.8 4.9 Plastic 100 Polystyrene mL volumetric pipettes flasks. 4.10 Miscellaneous laboratory glassware. 555.:.321 MOOarrgiinooenntMMioocddSeetlilr9EP6lAa09t9e40.00 ESxolpiadndStaabtleeCIoomnbiAnnaatiloynzFeolruoerqiudievaelleenctt,rode or equivalent. 5$3.:465 MOIirBciMroonscRoofSmtp2a3Etx2icbeillnet5e3.r8f0a6(coeonrcla4yb8ln6eeec(dooemndlpiyuftnueesreid(neogdnlOiyfruinsoeinend3geEdOrsifioufostniw3anrEge)sO.orfitownar3e) software). G0INTERFERENCES Q&LM1wsaterrecwiolmlmgeennedreadlltyhabtrtihnegpsaHmpbleeattoorpnHeoafr6.6.00.. A 1:1 mixture ofTISAB and sample/Milli 62&t:23rToShoaemmprtlaetemeptteehrmeaptseuarrametpularesesthmaeraesytsateinfrdfraeercdtdasftlwuseohrroieudlewdhmbeeenasctuohnreseimmseetntetent.r wwiIatitshctrahleeicbrroaamttemedet,hnedsetdantdhaartdtshewesraempslteimbede. 03914 2 000064 u6nddeArirelbeuctbrboldees. rapped under electrode can give erroneous readings. Make sure no air is.trapped 7.1 No special handling necessary. 8$8..1111.21PLMraeebpaeaslruarttheeio5fn0loamsfkLCsaoalsfi0bT.rI0a5St,Ai0oB.n1I,lS0ti5na,tnod15a.0r,1d02s0ndm1L.5pplapstmicF-v,oalluomentgrwiicthfltashks.date and your in l8a.b1e.l3ePdipfleatstkes.0.05, 0.1, 0.5, 1.0, and 1.5mLof 100 ppm fluoride standard into theappropriatiteilayls. 8.1.4 8.15 SAhdadkeaptphreofxliamsaktsetloym3i0x mL the osoflMuitliolnis-. QTM waterto each flask. t&hLe 6flaEslkismoinvaetreatiherabiurbbbulbebslefsr.om the flasks by tipping the flasks on their ides and rolling the air in 8.1.7 8.1.8 Bring Invert tahnedvsohlaukemethien ftlhaeskfslafsokrs up the tfointahlemi1x0i0nmgLmark with Milli-QTM water, 8.1.9 Record standards in Standards Log Book. 888...222..21CIaInflvnieebrcrteasptsriaoorbnye,troewmeotvetotpaspeealfrom electrode filling hole. 8.2.3 82.4 Ejecta Fill the efleewctdrroodpesowfitfhilfliilnligngsosloluuttiioonn.from bottomofelectrode to wet lower seal, tS8ho:em2em5catTniuhofena,cmteuutsrieenrrg'assntdianntsdhtearruFdc-stiewolniestc.htrcoodnecaernetrtaytpiiocnaslloyfc0a.l0i5b,ra0t.e1d,b0y.5d,ir1e.c0,t manedas1u.r5epmpenmtFwoitfholnloowbilnagnk t88i.:s22s.u76esCRleecaonrdthteheelselcotpreodien bthyeraipnpsriongprwiiatthe Mloiglblo-okQ.TM water and wiping the sides down with lab 88..33.1SCtaolriabgreatCioonndsittanidoanrsdsfoarreSsttaonrdedaradtsroom temperature. r99-e.11c.o1CmaTmlhieebnrdsaeatmdipoblnyeattnohdebTeMIemSaesAausBrumreaemndeunfntaec,etduSsrtetaron.bdearmdixmeedtwhiotdh:TISAB using the proportions c99l:-e11c.23trAPolldlaeociwesatihnsseteirrstbaedam,rplmineatkthoeemssiuaxrmepftloheearraenfdaerwpelsnaecoceoaitnrhdesbusbbaebmflpoelrseetiornnaspetprhetedinsugtnrtdhpeelraettlehe.ecterloedcet.rodWeh.en the 9:14 same The rate assamthpelecaslhiobrualtdiobnestthaendsaardmse. temperature as the calibration standards and stirred at the 91-518 WhenWhen the readinggs have stabiliized, zed, recrecodord thethe readireadinng g iin n thethe apprapor priate log book 03915 3 0000675 9.2 Calibration AndMeasurement, Using Orion 3E Software: 99..22..11.1CaFloilblroawtisotnep:s 8.2.1 to 8.2.4. 9.2.1.2 Press Function Key #8 (F8). c9.o2n.c1e.n3trTahtieocnoomftphuteestrasncdraerdesn,wialnldaswkheytohuertoorconnoftiarmbltahneknius mtbo ebre of standards to included in the beused, Make any necessary changes to the information presented and click on calibration. CONTINUE. s99t..a22b..i11l..i54zeOPdbl,ascpeerrevtsehsetAehleCerCcetEardoPidTnegisRnEotnAheDthfIierNsgtGrs.atpahnidcadridsoplnatyhoensttihreplcaotmepuatnedrc.liWckhoennCtOheNTreIaNdiUnEg.s have 92.1.6 Repeat step 9.2.1.4 and 9.2.1.5 for the remaining standards. i9n.t2e.r1c.e7pAtfatnedr the final correlati sta on. nRdeacrdo,rdthtehceosmloppuet,eirntweirlcledpti,sapnladycotrhreesllaotpieo of the curve, n in the appro as well priate l as og the boo k and click on CONTINUE. The calibration data is automatically copied to C:\Orion\Data\Calib.txt. t99o:.22s..t22o.r1DeaSdtealateacSitpnr.ceitahdesrhNeeEtW: or OPEN from the FILE menu to open a new or existing spreadsheet 9.2.2.2 Record the nameofthe `spreadsheet used in the appropriate log book. 9.2.3 Fluoride Measurement: 9.2.3.1 9.23.2 EFnotlelrowt:hsetneapsme9o.2f.t1htehrsoaumgphle9.i2n.4the appropriate place on the screen. 9.2.3.3 Click on the NEW SAMPLEbutton a9.p2p.r3o.p4riWahteelnogthbeoorke.adings have stabilized, click on the RECORD button and write the result inthe 10.1 Quality Control: 10.2 Precision and Accuracy 10.3 ppm Other Validation Parameters According to fluoride up to a saturated solution of fluoride. Reference 13.2, the rangeofdetection is 0.02 111 Calculations None necessary. 11.2 Analyzing the Data None necessary. 2.0 ATTAC HMENTS one 3.0 REFERENCES 02916 + 000066 I1n3c.o1rpOorriaotnedM,od1e99l1.EA940 Expandable fon Analyzer Instruction Manual, Orion Research R2es:eaOrrcihonInMcoordpeolra9te6d0,901099S1o.lid State Combination Fluoride Electrode Instruction Manual, Orion 4.0 REVISIT Revision Number ReasonforChange Revision Date 03917 5 000067 3MEnvironmentalLaboratory Method Extraction of Fluorochemicals from Rabbit Livers SOP Identification Number: AMDT-M-4 Revision Number: 0 -_-- Adoption Date: 4-(-95~ Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: Gres) ew roup Leader Li Fak Quality Assurance [74424 Date: ids Date Software: MS Word, 6.0 MAfafsesctSepdecDtorcousmceonptys.: M-5, AnalysisofRabbit Extractfor Fluorochemicals Using Electrospray 03918 0DOO0GS 1 1.1 eESltcehocyptler:aocsTepthraaitysemmiaestushsosddpeticosterfxoorstcrtoahpcey.efxlturoarcotcihoenmiocfafllsuroroocmhehremivcaelrssfroormaanbailyTsveer 11.32 MAaptprliicceasb:leRaCbobmipoLuivnedrss.: Fluorochemicals or ther fluorinated compounds. 20KE2.Y1 WOcFolRmupoDoroucSnhdes_m,icea_xltsra,_crtaibo_bni.t_liv_ers,_cle_cur_ospr_ay m_ass specwrometer, Foor 3.1 Use gloves when handling the rabit vers, they may contain pathogens. 4.0 SU4.P1PLISuEpSplAieNs DMATERIALS 44..11..21 4.1.3 GESlpyropivenengsdeo,rcfaptaybpleeoorfmdiesapsousrsbilneg1pi0p0etusl 44.:11.53 4.16 PPllaassttiicc Labels cgernitnrdiifnuggetutbuebses, 15 mL 44.11.87 TNiimneorgen 444.-.111..191110DFAiimlstlpeyorsis,acbTaillteappnilpnaesystli:ocng3ascyscrisvnygoreilnfgiuelstme.res, p0i.p2etun.. 4.1.12 Crimp cap autovials. 4.2 4R.e2a.g1enPAtqpsumeoaquuseAoumsmsoonluituiomn oAfcaemtamtoen(iAludrmicahc)e,taatpeprboyx.ad2d5i0ngpp2m5:0 mPrgepmareaa 25s00 4.2.2 saocleuttaitoen. 100 mLs Milli-Q water. Dilute this solution 1110 fora 250 Sodium carbonate/Sodium Bicarbonate Buffer (IT. Baker), pp ba(rnNidn,g2C1.O0YvNgoSlouHfmCseOodw,ii)tuhm0.Mb2Ei5lcaMlr-:bQoWnweaatiteegrh(,N2a6H5CgOo,fisnotdoi:u1mLcavsoblouimteetne 0fa0a)ond 44.:22..43 4.2.5 EDtihlyulteAcaecettaotneitrile Methanol solution, dilute setonitle 1:1 with Mill-Q water. 4.2.6 Mill-Q water 5$5:.21 53 VCUoelrnutmaei-xtTuumgrierxaexrT25 Grinder for grigrninddiinngg Iver samples 54 Shaker 03919 000069 2 5.5 Analytical Evaporator 6.1 There are no known interferences at this time. ZOSA7-M1PLTEheHraAbbNitDlivLersIaNreGrec_eive_d fr_ozen_, and performed. must be kept frozen until the exwaction 1s 8.1 8P.r1e.p1arapPetrreitpolanuroeoroafcntIiannntteeesrrunnlaapllhsotnSaitncadanacdridadrotfdoaspbepraodxdiemdattoelcayc1h2lpivpermsa1Hm,p1lHe.,2H,2H8.1.2 WReeciogrhdatthleeaacsttu0a.l1wegoigfht1.H,1H,2H,2H-perfluoroctanesulphonic acid. 88..11..43 DwBairtlieunrtg.eiCt3aulmpcLutlsoaovtofelttuhheemeascttwouicatklhsvt1aa0ln0udemaorLfdttohoef2m5set0atnhmdaaLnrod.lfi,natlhivsoilsutmhee swtiotchk Msitalnldia-rQd. actusalulmphgonpiecrfalcuiodroctaneorL X 235m0L mL = actual value, ppm 8.2 8P.r2e.p1arPereFpCar-e9F5C-A9n5iosntanSdtaradnsdfaorrdtshe standard curve. 88..22..32 8.2.4 BDWireliiungtgehutaphtpeposroovlxouiltmiuaomnteewlwiyitth1h0d0diilmultugeteaocafecteFotnoCin-ti9rtir5lileaen1i:dn10r2feoc1or0r0damtsLhoelvuaotclitouunmaelotfwreaiipcgphrftol.axs.k.100 `appppmr.oxD.il1u0teptphmis.solution1:10with dilute acetonitrile foar solution of 8.2.5 U5.s0epiphm1,01f.0opmpsmolauntdio5n0t0o pmpabk.e working standards with values close to 8.3 P8.r3e.p1armWeeLisBgeMheifl4l0Liig-vQoefwraBtoHevro.imnGoergilenivdneartotineatohtoaom2o5Ug0semenLofuosNraslSoglteuatnnieodnba.ortdtlse containing 200 8:3.2 A1 dmdL1amlLiqouoftstohfe tshoelustoilountitoonai1n51m5LmcLencternitfruigfeugteubteu.bePsr.eBpaeresuareottaol ofcight 8.3.3 Ssrtpeaisnkudesaprsedensvdeinsnooslftuettpihoe9n.11b2ymoLsfhaatklhiieqnupgortoistcbewediuttrwheet.ehnOenafleoilq1luoomwtsiL.ngalaimqouuotntsserovfeswoarsktihen.g blank. C39%0 000070 3 upgrFoCx-i9m5a/tgelivCeornc.|| (WoArpkpirnogxiSm@antdeard iC Conc.) --pak4 8.4 Calculate the actual valueofthe standards: ulof standard x mg/L x 1L/1000000uL = mgFC-95 rT37omgDliEveErC/19mSL_s_olutixon 45 (organic layer recovery factor) = megqu/imvaLleFnCt.s9i5n liver mgmL x 1000 = mgL = ppmFC-95in liver r*eApvoerrtaedgeasweeiqugihvtaolefnbtosvoifneFCl-iv9e5r ipnotsoalsustiiounm.sTalht,e amountofFC-95 is 8.5 8C.a5l.i1bra`Emtxetitrohanocdt.tUhesesptihkeesdebesteafnldiavredrshtoomeostgaebnlaistheyfooulrlocwuirnvge9o.n13thteo m9a.s2s3.ofthis spectrometer. 8.6 8S.t6o.r1ageNcoevCweornsedtdaipntldiaasortndissc acferonetrprriSeftpugaaernedtduabwreidstshunetialchthaenaalnyasliyss.isSotanntdhaerdmsasarsesspteocrteodmienter is performed. 8.7 S8.t7o.r1a"geBeeCfonldivietrihoonmsogfeonratSetsamnadyarbdesfrozen after preparation. 9:1 pOabctakiangefdrowzietnlhiovtehrersatmipsslueess..In spent tissue, note that the liver has not been 9:. 3 Usceigahditshesescatminpglescdailrpceclrlaynidntcoutaotfareadppprlaosxtiimcagtreilnydi|nggotfublei.ver, 9.4 Record the liver weight in the study note book. 03921 000071 4 9-8 Pinuittiaalsl.abel on the vial with th study number, weight, rabbit ID, date and analyse 99..67 mGAirdniduntd2e.st5,hemunsLtaismlplwteaht.eersP.aumtptlheeigsr2inhdoemropgreonbeeoiunsthseolsuatmipolnewiatnhdngorilnadrgfeorchaubnokust.2 29-:98 TERPai-nk2see2t.thheegprrinodbeerofapfaritntaontdheclseaanmpilt ewiwtihthme2t.h5amnoLlsawftaetreeraucshinsgamaplpei.peF.ollow AMDT- 99--1110 iPCniafppoetrtmh1aetmisLoanmipanlsteothaaen1gd5rmvionLrdticenexgnttfruobiref.1u5g(eSseetecboeAn.dMsLD.aTb-elMt-h4e cWeonrtkrisfhuegeettufobredwoicthumtehnetiidnengtitchael 9-12 rSpeimkaeintihnebgesetfelpsi.v)er homogenates as described in 8.3. with the appropriate amount of FC-95 standard 99-:1143 AaScpdeitdkateIe.tmh9eL.s1oa5fmUptshlieenssgoaadnnidbuaemneacflaylrtibivocenarlatpheio/pmseootgdeianudamdtbSeiscmawlribtoehntah1yt0le0abucuefLtfoaetfre.ainntde1mamli.staanmdmaordn.ium 99.-1165 PCuatptthheemsianmptlheesahnadkevrorftoerx2200mtion. 30 seconds. 9-17 oCnentthiefcuegnetifforug2e01100.2255. minutes, uniil the layersare well separated. Set the power 9-18 9:19. mBRleLomwogrvtaehdeu4astamemLdpslgoelfatdssohwepinpteootpn.oTtrrhgaeannasinfacelryltatiyhceearll0aebavealpftoroersaththeo1rf5rtmeosLnhectaeurnbetd.rriyfnuegses twuibtehwniitthroage5n, 59::2319 aBrpipnrgoxtihmearteemlayi3n0intgos4a0mpmilneutueps.in 1mLdilute Vortex 15 seconds. acetonitrile with an analyrical pipet, 9-22 Ttshrteaundsysafmnepurlmtebheeinrstaoamnapdlfraeenstihomcaael3ntnmruimLfbuesgrye.r.tinugbee.orAtataacuthovaia0l..2 nLambenlyltohen tmuebsehorfivltiearl, waintdhftihleter 99..2243 CCoampplaendtehoAlMdDfTor-aMn-al4yswiosrbkysheeleetctarnodspartatyacmhastospsapgeecorfossctoapdyy.notebook. VALIDATION 1100..21 QPrueacliistiyonCoanntdrolAccuracy 10.3 Other Validation Parameters 11.1 None 12.0 A12T.1TAWCorHksMheEetNATMSDT-M-4 13.1 AMDT-EP.22 Revision Number ReafosrChoannge Revision 3 Due 03922 000072 5 Worksheet AMDT-M:4 LE ee EE poe tr rpg -- rr rr rr rr rr _-- -- r r r rr bee e---- LL ---- | r er e] Fer {PierTol ofTverSlip Ee Er --- Plomaiiee ------------------ TcAdotak | ---- | cEF-- mm [Roe snlo EbAgere | PE osEeRtE iee--------------e ---- e] | (hake Opp---------------- | | _-- EE a E---------- Poser -------- mm mmm | 03923 ` 000073 3E M nvir Labo oratn orymental Method Analysis of Rabbit Liver Extract for Fluorochemicals using Electrospray Mass Spectroscopy SOP Identification Number: AMDT-M- Revision Number: 0 eee Adoption Date: /--( 45 Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: roup Leader 4 : Quality Assurance Les Date Lh fee Date: Software: MS Word, 6.0 Affected Documents: M-4, ExtractionofFluorochemicals from Rabbit Livers 03924 000073 ? 1.1 Sifslcupoiepdresf:ofrTorhmiefsdlumboeyrtoshcihonedgmlieisciafolonsrmutosheniinagtnoatrlhiyensgiesloeofcfFterCxo-ts9rpa5rcataysnimooafns,rsaMbsb/pieZtc=tlri4vo9emr9e,toertrho.etThihentreeiransnasaluleyssiosr 11..32 MsAtapatpnrldiiaccreadsb:Mle/RZaCbo=bmi4tp2oL7i,uvnaednrsds:(ostFahmlepruloaerpsop)cr,hopeBrmeiieacftaeLlismvaeosrrseo(sst.thaenrdafrlduso)r,inoattheedrctoismspuoesunadnsd. fluids. 20KE2.Y1 WOFlRuorDocShemicals, fluorinated compounds, electro~spr~ay~mass spectroscopy, mass. spectrometer, rabbit livers. 3.1 3.2 iDUnosteno octthaeurtpuirnoonbtheweiDtphOutmhNpeOvaToblotTvaOegeUitcC'asbHclaeTpfaHocrEittPyhoeRfpOrB4o0Eb0e,.0 tWphsheir,eenIifstprhrieessvkosoulfrteealgegceotcerasiboclaveleirssh4po0cl0ku.0ggpesdi. 3.3 fsitnodp tahnedprleulgeaasnedprreepslsuarcee.tThehpelpuegagkedtutbuibninggm.aSyeebAeMpDluTg-gEedP.-T1r5oubleshoot back to Donot run the pump to dryness. 4.0 SUPPLIES AND MATERIALS 4.1 4S.u1p.p1liNeistrogen gas regulated to 140 psi. 4.1.2 4.1.3 F1l0u0ofuiLx ocrol2u50mnuLo.rfelqatuitviaplseynrti.nge for sample injection. 4.2 44R..e22a..g21enMDtiisllultie-Qacweattoenri,tialllewmaotbeirleuspehdasien,tdhiislumteetahceotdonsithroiulled1b:1e wMiitlhliM-iQllwiat-eQrw.ater. $5..21 VISGCTOrSioyr2i0n0g0e EPluemctprospray Mass Spectrometer or equivalent. 55.:43 S10 pueL AcsstemabAlypS3h00yAustosiamcplser 5.5 Autovialsor capped centrifuge tubes. 6.1 There are no known interferences a this ime. 7.0 SAMPLE HANDLING 7.1 uKnetielprtehaedeyxftorracatneadlyssaimsples in capped 15 mL centrifuge twbes or in capped autovial0s 3925 0000773 2 80 CALIBRATION AND STANDARDIZATION 8.1 Preparation of CalibrationStandards 8.1.1 Seven beef liver standards extraction procedure. (See and one blank beefliverare AMDT-M-4, section 8.0) prepared during the 8.2 Calibration 8.2.1 Run the sevenbeefliver standards to obtain the standard curve. twice, starting with the lowest standard 8.2.2 iTnyptihceasllaymeonreansgteanodfarcdonicsernutnraatfitoenr aesacthhe5stamo7plesasm.ples. Choose a standard 8.3 Storage Conditions for Standards 8.3.1 pcFeorrvefesorhresmdteapdln.adsaStraidcmspclaerneetsrpiarfneudpgaesrtweadendwsairtudhsnelaarcethhNeaOannaTlaylrsyeisfsir.siSgoetnraantthdeeadr.mdassasrsepsetcotroedmeitner is 8.4 8S.t4o.r1ageBeeCfonldivietrihoonmsogfeonrateBseemfayLibveefrrHozoemn oafgteernpartepeasration. 9.1 Initial Set-up si 99..11..12 SReetcosrodftbwaacrkeitnog"pOrpeesrsautree ionn"t,heloinnsMtorudmeenEtS".log. 9.1.3 Fill the solvent cylinder with mobile phase. 9.1.4 cSeotmithnegpouutmpofttoh"eRtuinp"o.f Stehte pthreobfel.oTwhteo p1r0e0s0suurLe/msihno.ulOdbbseeravte1sd7r0o0ptloet1s300 9.1.5 Check the chips. The fused silica at tip should be the flat end of the probe. Use with no. jagged edges. an eye Ifany piece chips to check for are found ctuhte oapfpfrtohperitiaptoeftlehnegtshi.lica with acolumn cutter and pull the silica through to 9.1.6 Check your nitrogen supply. Tum on the nitrogen. There should be no. nisogen leaking around the tip of the probe. A fine mist should be`coming 9-1.7 Coaurteoffultlhye tip. guide the probe into the opening. Insert it further. Connect the voltage cable to the probe. nil it won't go any 9.1.8 `Gapoprtooprtihaete"Emdaistsoer"s ptaog4e2,7anadndse4t99I.onization Mode 0ES', and the 9.1.9Ifitis not in single ion mode goto Option and set SIR. 9.1.10Start Acquisition. Assign a file name, MO-DAY-YR + letter. Record iit n the log book. 9.1. 11bReugnintnhienbgeeoffltihveerru.n`.s.amRpulnes 510 7 samples. afiQrsCt, rcuhnenciknbgyearcuhnnsitnagndoanred sttwaincdearatdatfhteer every " 000076 9.2 Manual Injection 9.2.1 iDnrjeacwti1on50puorLto. fIn:jseacmtpslleowilnyt.o Raescyorirndget.heInsjaemctplteheIsDaimnplteheinltoog tbhoeorkheodyne 99..z2.32 WTauimt tthweovmailnvuetetos,"Oann"d.inject the next sample. 03926 9.2.4 Record the scan number for each sample in the logbook. . 3 9:3 9U.s3i.1ng StehteupAustaomsplaemptlraeyrA, B, or C. 9.3.2 Rvieaclosrmdatyhebesaimnpclaecsharnund. their positions in the instrument log book. Up to 17 9.3.3 9Se.t3-.u3p.t1he sampler: Push the sample button 99..33..33..23 SeSettsianmjpelceyslaomoppsliez=e2 = 100uL 99..33..33.54 SNeatmCeytchleetfiilem:eLi=v0ers 99..33..33..67 IAddedntitfhye sthaempwlaeysutsoeQdueue by pressing "Enter" 9.3.3.8 Press "Rutonsta"rt 10.0 VALIDATION 10.1 Quality Control ~~~ 10.1. 1spRteuaannkdaahsredtiagwhnitdllaorcbdeeueruvsseesrdt.yoTp5httehoe7rreusmnaa.mipOnlnielnsgy.Istfahemapssliaegmsnpilwfeiilsclabnbetefcorrheeaantnahgleyelz(aesd5t.0ac%c)epitnable 10.2 1Pr0e.c2i.s1iSoeneaMnedthoAdccVualriadcatyion Report number AMDT-M-5.0.V1 1100..43 ROetfheerrtoVMaeltihdaotdiVoanliPdaatriaomneRteeprosrt Number AMDT-M-5.0.V1 1111..21 PClaoltcutlhaetisotnasndard curve, using the mean of the two values obtained for each s11t.2a.n1daRrdre.egardesspieoankthoediegthetsrmoirnaertehaes sfoarmptlhee scaonmcpelnetsraftrioonms.the printout. Use linear 11.2:2Claivlecru:late the mg of FC-95 anion, or other fluorochemical in the total rabbit mg FC-95 anion in the total rabbit liver = mgmsgof iE vaenrioC unsfedr- ofomrsai9 nda.lcysu5 isr.ve x Total massofliver, gms 1111..43 MPraikneatacrhersoumlasttoagbrleamanfdoreentaecrhistaimnptlhee,swtiutdhy tbhoeokp.eaks labeled with the sample or ssttaunddyafrodldIeDr.. WSrtaiptleetahlel scthurdoymnautomgbrearmosntothgeetphreirntaonudt,niunimtbiea,rdpaatgee,sa.nd put it in the 03927 000077 4 1A 20 TTACHMENTS None 13.1 AMDT-EP.17 Revision Reasonfor change DRevaiseion 03928 000075 5 3M EnvironmentalLaboratory Method Analysis of Fluoride Using Ton tSheelecStkiavlearElSeecgtmreondeted Flow Analyzer With Method Identification Number: AMDT-M-8 Adoption Date: jos Revision Number: 0 --- Revision Date: None Author: Deb Wright/ Cynthia Weber Approved By: upLeader fa g g* 7 Quality Assurance 2/75 Date J-27-45" Date Software: IBM MS Word, 6.0 Affected Documents: AMDT-EP-26, Analyzer Operation and Maintenance of the Skalar Segmented Flow 03929 000079 1 L.1 TthheisDomhertmhoadnnisDfXor2t0h0e0an(aAlyMsDiTs -fMor-1fl)u,oraidned, ctohlelremcatleldyinexTtIraScAteBd fforroamnaslaymspilsewsituhsianng Ion Selective Electrode (ISE). The analysis is performed using the Skalar Segmented Flow Analyzer with ISE. 1.2 tShaempDlohersmcaannn.be tissues, serum, biological material,or other materials extracted on 20 KEYWORDS 2.1 Skalar, segmented flow, fluoride. 00000000 4.0 SUPPLIESANDMATERIALS 4.1 Supplies 4.1.1 4.1.2 ASuatmopplipeectusp,so,x4fomrdLorplaesqtuiicvcaulepnstwwiitthhcpalpasstic tips 4.1.3 Polypropylene volumetric flasks, 100 mL. 4.1.4 Cartridge components, refer to the Skalar Methodsfor components and part 4.1.5 Snaummbpelres.prefilters, Evergreen 4.2 Reagents 4.2.1 Brij 35, 30% S.F.A.S. Detergent 4.2.2 `TTIISSAABB IIlI abudfdfe2r.5somluLtioonr: 1P0u0rpchpamseflTuIorSiAdeBSIoIluftrioomn Oarnidon1.mTLo B1rilji.ter of 4.2.3 Samplerrinsing solution: Dilute TISAB II 1:1 with Milli-Q water. 4.2.4 Nitric acid solution for decontamination, 1 N (lab grade): Slowly add 64 mLs concentrated nitric acid (HNO)10 250 mLs of Milli-Q water.Bring the volume up to 1 L with Milli-Q water. 4.3 Standards 4.3.1 Stock solution, 100 ppm F: purchased from Orion. 4.3.2 Intermediate standard, 10 ppm: Dilute 10 mLsof stock solution to 100 mLs 4.3.3 WwiotrhkiMnilglis-taQnwdaatredr:.MUaskeepoulpyptrheopyfollelnoewivnoglwuomertkriincg fsltasaknsd.ards by adding the `vooxlfuormdesoropfuimntpemrametdeiaptiepeosr,sttooc5k0smtaLnsdoarfd TiIndSiAcaBteadnodnditlhuetitnabglte,o using 100 mLs [Workoio0np igSpSm teamnd-- a1 rd TmL ofSto- ckSnead ls of1 tegrmpe5disa--te --Sta--ndard with Milli-Q water. --odoospp ppm mm-- ---- 71 03 p 5 = 15---------------------- 039300 0000)50 S.1 Skalar Segmented Flow Auto Analyzer SansTM System equipped with ISE 6-1 Hinitgehrfceorenncceensu.ations of alkalinity, chloride, phosphate, sulfate or rom cam cause 7-1 Swiatmhpilne3s0sdhaoyusl.d be stored in polyethylene bottles. Samples should be analyzed 8.1 8P.r1e.p1araPtriepoanreocfalCiablraitbiroantsitoanndSatrdasndasaridnssection 4.3. 8.2 8C.a2l.ib1raTthieonstandards are analyzed at the beginning of the run. 8.3 8S.t3o.r1a"gesSttaaCnnoddnaardrdidstsiaarorenepssrteofproaerrdeiSdntacatanapdpmaeirdndpsiomluymporofpyelveenrey vsoixlummoenttrhisc,oflraskass.nNeceewssary. 9.1 9St1a-r1t 9-1.2 UCplaPmrpodcoewdnurtehe pumpdecks, air bars and Put the fluoride electrodes in the electrode schaammpbleerr.-pump tubing. 9-1.3 Tbauthm on the power of the sampler, pumps, offset potentiometer and heating 99-11..54 TtPuuurtmnthoeonnretahtgeheeinnttilenirtnfeaercsfea,inccetohmebpeaufptpoerrroe,prdtiihasetpelcabooytmatplneudst.eprri.nter. Make sure you 9.1.6 Let the system stabilize for approximately 30 minutes. 9.2 9St2a.r1tinCgreaateRuansample the name of the ftialbe,leanbdy sperleescstEinNgTFEIRL.ES, TABLE, and CREATE, type in 9-2.2 9.2.3 PPrRiInNtTt,heGsRamOpUlePta1.blTe,hiisnsweirltledpinntthsehesymsntieme table by Dial the sampler scrtings to the appropriate numberof pushing ESC, samples, number of 9.2.4 Fsielclotnhdessfaomrpslaemtprlaey FW/RUNOUT cups wwaisthh,thaendstnanudmabrdesr, on the sampler do osfasmepcloen,dswafosrhetsheasnadmpdrlief.ts. not need tobe filled. IW and 9.2.5 Set the baseline. 03931 000051 3 9-2.5.sm1eaSleyelcetbcietngiGnCRtOAhePNHDTaIRtCaOSHL,anPRdAElNiAnELgLTPaIannMedEl..puSIswfhiyitoncugh cFta7o.ntnhoetAgnaelyrseiasl-tPiamnee,lypoyu 9-2.5.tb2haUessebeolittnthe.eomsAmodajflultshtsectsrhceerwebdearnsi.evleirnefournttihleiotfsfsaetppprootxeinmtaitoemleyte3r/4toisnecthtfhreom 92.5. 3thCehienctekrftahceehsicghreeswt#s3t.andard and adjust the gain, if necessary, with 9-26 0GwiolrltemootCtObtoeNrTtuhRne.OA(nLSaelPlyeAscNitsEoLrPa,dneeaslen.ldectot aannaallyyssiiss pbayneplr.esDseisnegleEcNt TthEeRa)nalPyrseissstThuatb 99.-22..87 9:2.9 PSerleescsttShe sTpaActeboRasrtaTtrotbrthienganuaplytshies.local menu, Type your ID Ginitals), the sample table which you created under 9.2.1 (or aprnedssseEleNctTSETRAfRorTcAhoNiAceLsY),SIchSo.ose running with or without the system table 9-2. 10SAsfKti.teor tthaetriignhgttnhuemsboefrtwoafres,asmtpalrtesthaensdatmhpaltert.heMsaakmpeles/uwreasthhiaaitrhetismaemspalreer is 9-2. 11Select GRAPHICS, REAL TIME 10 view the progress of the analysis. 9.3 9L3o:a1dingaGnodtasonedlCecOPtrNLiTOnRtAiDOn..g SPteAhleeNcEtDLaA.tUapT-rFOeisClseAtLheCsUpLacAeTbaIrOtoNbarnidngenutperthtehelofciallemnaemneu(or 9.3.2 Thioghvliieghwttthheecfailliebrtoatbieonprciunrtveed, CURVE. ganotdoprGesRsAEPNHTIECRS),.CALIBRATION 99:.33..43 TToopprriinntt tthheelhoigwhlelveevlelsccurerevne,, SETTINGS. Change the max ppuusshh EPSRCINtTo gSeCtRoEutEoNf.graphics. Select y value to approximately 900. Go to CAL 9.3.5 pRCrUeesRswVEENtoTaSEnEdRTpTarnIedsNsGPESRSICaN,nTdacSnhdCaREnngEteeErtN.hePtromepasrxsinPvtaRslIauNempTblaSecCkpRetaEokEs4.0N9,5, go to EDIT, 9-3-6 OToUTprPiUntT.theSerleesculttsPRgoINtoTCERONfoTrRthOeLEpPsAoNnEoLr,PSRPNACfoErBtAheRL,asOerU,TPUT, 9.4 S9.h4u.t1dPouwtnall the reagent-lines in Milli-Q water. 99.44..32 oALffefttseetrthtephosetyessnyttsietomemrmeitnhesares.frToirunsamepdporfcofoxmtiphmleaehtteeealltyyi,n3tg0ubmnaitnohuftofentsh.weeseakmepnldesr., Lepauvmpe alinqduid in 597:34..54 RTtehalekeelaisnteehse.tehleecptruomdpe-doeuctksa,ndaisrobaakrsinan1d00sapmppmleFropvuemrpn-itguhbt,ing. 9944..76 OSenleFcrtidFaIyL,EtSu,rnproefsfstAheLcToFmpuatnedr,sednistplQaUy IaTnd10inetxeirtfatchee fporrogtrhaemw.eekend. 10.1 Q10u.a1l.its1yiRgunCinofniatcsartnoatlncdhaarndge(miidn ptoeahkighheicgohntceonctcruartsi,oonn)leyvethreys1a0mpslamepslebse.forIefathe last acceptable standard will be used. The remaining samples will be reanalyzed. 03932 0000a82 10.2 P10r.e2c.is1iSoeneaMnedthoAdccVualriadcatyion Report number AMDT-M-8.0,v1 1100..34 ROetfheerrtoVMaeltihdoadtiVoanliPdaatriaomneRteeprosrt Number AMDT-M-8.0.v1 11.1 LCLa.lc1.ulIaTthieonstsandard curve is plotted by the Skalar software. 1111..23. 11.1.2Abeltltecr.alculationsaredone by the Skalar software. 2 `should be 0.995 or WParietpeartehespsrteuaddyshneuemtbsetroosnumtmhaerpirzientdoautta,. Include sample volume, weights used tc. oS`nwuittmohmsaaltrluypdayschdkeiaesgtkeaandndodctbuaampceekniutnpstodatinshdkesps.ltaucdeyinnottihenetbiosaotlk,u.dcyaBtfaeocltkdheeru.pprMailanltkdoeaust4.acsaonnpdysopfcrsettahodegsehetehtesr 11.4 1El1e.c4t.ro1niGcLDPatsauwdies: Electronic data 11.4. Study, and also data is copied oisntcoopaiefdloopnptyo dtihsekStthuadtyi fsltooprpeyddiinskhfeorJocra.ch 20ther studies: All data is copied ontoa floppydisk tha is stored in he Jab. Loarracements 00000 13.1 13.2 'ADMXD2T0-00M-O1rg,anTihcerHmaallidEextArnaacltyizoenro-fLiFvleuroride by Means of a Modified Denman Skalar Methods, #335, Skalar Methods Manual 13.3 AMDT-EP-26, Operation and Maintenance of the Skalar Segmented FlowAnalyzer ReLvibsiooRn evisions 0000 000000 Number Reasonfor change. Revision Dae 03933 000083 5 9.3 Quality Assurance Unit Statement 03934 000084 AuachentD GLP Study Quality Assurance Statement Study Title: Single-dosTe-6D0e5r3mailn RAabbsboripttsion/Toxicity Study of Study Number: AMDT-022195.1 NaofAmuditeor: Kari Rambo TThhiesfsitnuddyinghsawsebreeenreinpsopenceted0d tbyhetshheaQduyaliitrycioAsrsaunrdanmcaenUangietramseinnsdicatiend the followinggatabblee. Inspection Dates From To -- Phase 10/1495 11/03/95 Final Report Date Inspection Reported to Management StudyDirector 1/0395 11/0395 A 7 #395 QAU Auditor Dae 03935 0000835 1 9.4 Key Personnel Involved in the Study 03936 000056 3M EnvironmentalLaboratory Key Personnel v `Thermal extraction followed by analysis using Orion fon analyzer: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Analysis of liver extracts using electrospray mass spectrometry: Jim Johnson Dave Christenson `anTahleyrzmearl weixtthraicotniosnelfeocltlivoeweedlebcytraondael:ysis using Skalar segmented flow Jim Johnson . Deb Wright "Rich Youngblom Deann Plummer Documentation and Reporting: Jim Johnson Rich Youngblom Quality Assurance Unit: GCyanltehViaanWeBbusekrirk Kari Rambo 03937 000057 9.11 Data 03938 000085 d9.e1t1e.r1miSnuemdmbayrtyhearnmdalraewxtdraatcat;ionugfoFl"lionwewdhoblyeanliavleyrsiass using Orion ion analyzer. 03939 000059 Summary of Combustion Data - Liver AMDT-022195.1, EWI 6329-137 As Referenced in Final Report section 6.0 DATA ANALYSIS Total ug Fluoride in Whole Liver Mean per Dose Group* Control Group 10 mg/kg dose (T60S3)* 125 mg/kg dose (T6053) 250 mg/kg dose (T6053) Bg Std. Dev. 20.2 + 7.9 18.5 + 8.7 16.5 + 4.0 17.2 + 6.6 *"C*aOlnceuloautteldiearsntohteimncelaundeodfitnrimpleiacna.te samples from each of three male and three female rabbits, 03910 000090 - Feao % pAoctmuFalnlver ApvpemraFg-e Liver iniver bumed Wfhiovelre ToitnwahoFl-e weight Iver SPKE3D 1 y 95% SPKEI2 oe% 0w4m0 0012 ww) G0.a15m2s5 Gam Go) 0.1503 SSPPKK11228812 8821%% 2117850 LIVER BLK 1 0373 00..1131033 01216 LF5I3V4E4R2B.1LK 2 Fsss22 00326655 00.111446s0 0231 0262 01132 wes 2075 FF5533446442-.13 Fs34642 00..119840 00.1125490 0170 0175 01447 857 1517 FF5533446545.-31 Fa3ss.2 00.211602 00.11232735 0145 0178 01588 w331 1657 lFi5v3er45b5la.n3k-1 liver bank2 00.41737 0200 00.1110298 0.1378 lvveerrssppiikkeot2 9903%% 01..023610 lverspike-3 90% 0.850 00..11412400 0.1440 FF5s334441i-21 F534413 00237919 0232 0301 00.11433858 11412 35 01343 FsFS33445s11-21 F551 00221315 0.181 0200 00.11135928 8841 01413 1847 FF5s3a4d6s1i-12 F53461.3 00.116953 0.168 0175 00.1125572 7211 01316 1273 FFs533446600-.12 Fs3460.3 00.210822 0181 0188 00.11327300 ss 17.03 0.1285 ilivveerr bbllaankn-2k1 00261074 verspike-1 90% 1.013 00..11336218 0.1342 iivveerrssppikkee2s 0673%% 01.847550 iverspiked 95% 1.397 00.11001817 0.1028 lFi5v3e4r4b7a-1nk F53447-2 00226353 00.11501087 0152 0185 01376 7908 1480 FFss3a4a4s72:-31 Fsus22 00114730 00..11126915 0152 0147 01159 042 185 FU5V3E45R2:B3LK 1 UVER BLK2 00.210462 0131 000...111635208280 LLVVEERRSSPPKKT2 1s020%% 11017248 00..11420420 Page1 Dosage (mary 0 0 0 0 10 10 10 RPTISTLXLS FSCP9I5KECDOAPTIAEDFFORRO6M3 A63N2D91136216.PPM DEONWDI4N-G5a05C 03941 000091 -- - Foon FasItD FF5s3s4d6s3.332 FFs533444444.12 FF5533444484.13 FF5534438.82 FF5s34s46e-21 F5F3s435434.613 FF5s3a445s33.23 FFasa3a4sse6t2 FF5533445463..31 F5F3s4344433..32 FF5533445500-12 F5F3s3445607.-31 FFs533446677..32 FF5533443309.12 FF5s43444350.13 FFsSa4s4a4s523 F5F454s4050.-21 FLi5v4e4r5B0l.a3nk-2 LLiivveerrSBpliakne-k1a LLiivveerr SSppkikee2 LLiivveerrbSlpainkkes4 FF5s33444490-21 Fs3440.3 PAcMtEua:l % inlver rovy 0W6W%) 01187609 00068280 00..116838 00118446 00.114807 00.214853 00218455 00156030 00.117950 00.117954 00.117614 00.212523 00127083 0011139 00230900 00214521 00115400 00..413422 83% 01..104988 8910%% 01..913429 96% 01.231078 00127438 0.163 * Outer not included in average. ApvpemraFg-e Liver Wlhiovelre iTnawlhoFl-e iniver bumed weight ww) @amy ams) fiver () 0433 00.01125825 8140 3528 00..11125803 0267 00.1133498 6027 1985 0172 00.11129592 7233 1247 00.11115659 0170 00.11311624 6278 1407 0225 00.11417998 0451 2130 00.11153133 028 00.11303012 7870 2184 0fs1 001.243681 Teds fagr 00..11300823 0163 0011027120 7689 1253 0201 00.11038875 T7586 1581 00..11308046 0216 00.11431298 81.71 1783 01e8 001.1236210 9581 1887 00..11437294 0144 0011217737 T7826 1128 00..11322585 00..11016433 00.11141606 00..11028749 ofe4 00.11245975 7080 1378 Dosage (mokg) 10 10 10 125 125 125 125 125 125 250 250 250 250 Page2 RPT137LXLS 03912 000092 Feog % F535i04D-1 roy F53504-2 F53504-3 F53456-1 F53456.2 F534563 F53454-1 F534542 F534543 Liver Bik-1 Liver Bik-2 Liver Spike-1 94% Liver Spike-2 Liver Spike-3 108% 104% Liver Spike-4 94% Actual PPMFnliver 0W.W13)8 0141 0.168 0.188 0730 0220 0.128 0A11 0215 0.146 0.140 - Average ppmFinliver wi) 0148 0379 0151 Liver bumed (0g.r1a2m7s4) 01368 01116 0.1017 01398 0.1258 0.1304 01414 0.1245 0.1149 0.1029 0.1074 0.1378 0.1052 0.1200 Whole TotalFliver inwhole weight liver (grams) (ug) 8231 1222 77.40 2025 9304 1402 RPTII7LXLS Dosage (mglkg) 250 ReplacfeormFe54n45t4 250 250 ReponlDaay2c by Fe535d04 03943 000093 Page3 9.11.2 Raw data; analysis mass spectrometry. of liver extracts using electrospray `This data, although supportive, in the opinion of the Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. 03944 000094 - I -- ----WREUET 6(332299-3; 7 (48)RofT I|E041005 5m i,2) | | ! a |i I |i RE ws 20 _-- . af 1 FewplesSreened be Ex sz" 250150 8t 34 4Bude io n oryngmeskRm RoBt cEriamaanncrs. ) | ]&ww. st51510 S270 ia &S Pr le 22 iizz |i || I Je wl wl ge, j f eT io | 1500 | |i 20 ii a ase Z2e7 e 1 a" |ii a ssi 1 | ios zu 200 20 a Rott Gs. |l 5P5 e| wo i|| sist est| mn ||i [ha 0Woam w m0T seoaig de sg am S|| 3|oonsasmn20 my smoot || | JT> ee ar 1541. rnsstn 2 [a i al a i| Is re Pm || : fg wo 8 | orlsithTorii |T oo TF 2 | isB| Se | cPit Brod Eg ~.a-- < J[onessm(40,209) i E | || | 220230 | sont 3501gp 12200 4083 SotsE ChametS ss.| 25 00 gp 47 TO a3sorstan | sm | rE =o % 1g i| Journ55 : || werie | i} | bi 0 wy og |[I fh 18]i & m wr = Siege | P soo oiIl , ag 3iaTom||| iow aEw Zfa2 fF i Tm 58 | a un - || bi0 M00 2600 2000 T3000 3200 400 3600 3800 4000 4300 %doq 4600 4800 5000 5200 don sao d9.e1t1e.r3miSnuemdmbayrtyhearnmdalraewxtdraatcat;ionugfoFl"lionwewdhoblyealniavleyrsiass using Skalar segmented flow analyzer with ion selective electrode. This data, although supportive, in the opinion of the Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. 03948 000095 - RE:6329-137 LIVER SAMPLES AMDT 22195.2 ADnaatleysotf:AnDaOlyWsis: 5-2-95 QA "20a onQube The samples are burned in the Dohrman at 958 C using between 8.1 and 0.2 grams of the liver. The gas is collected in 1.8 mL of 1:1 TISAB/Milli-Q water then an additional 1 mL of 1:1 TISAB/MIlli-Q analysis. The is added samples to allow are then for sufficient volume analyzed on a Skalar for Skalar Segmented Flow Analyzer using the lon Specific Electrode (I1SE) Method. TISAB buffer system. The issaamdpdleedthtoeneagcohessatmhprlouegahsaithepartoecdeemdisxitnhgrocuogilh the 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 8.015 - 8.15 ppm high and 8.15 - range was 1p.l5o0ttpepd musfilnugortihdee. inTvheerssetalnodgaarrditchumrvoeptifoorn.tThehe standard curve for the low range is linear. All standards and samples were then calculated by the Skalar softwareusing these curves. All results below 0.8861 ppm appear on the raw data as #.##4##. A quality control standard was analyzed every 18 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. QuaQuit 03949 missed afagebev D0009FT Ri J) 24 BEST COPY AVAILABLE SUMMARY of 6329-137 LIVER SAMPLES AMDT 22195.2 SA 42073 ] i a FsM@22 2ND FFo4i2l3 NNDD FFM466t32 NNDD FFssHsssl2 NNDD DosGeRLOevUePl:1 0 FFseHlssl3 0N0D3 FFSlMAaL2 0N0D2 FFMssiii2 NNDD FMsl3 FsMell NNDD FFs6M16l32 NNDD = 20 oo u: e NEDD) = ND a %06% a ND & AND 2200 00112%0 NNDD 0866756739 NNDD 2200 001347 NNDD ND 8se5st7y NNDD ND 2200 001178s NNDD ND 2933300%0 NNDD ND 2200 oounxs 0ND H5e3l0le%s dNsD1 2200 001183385 0N2D3 02 l1e4l1e6ts 2N6D1 237 2200 o01u3s% NNDD ND 88884400888 NNDD ND 2200 00132 NNDD 77400868 NNDD 2200 001767 NNDD ND 2M770044 NNDD ND EF3H60L2 NNDD 2200 0OB1W0 NNDD ND ssissit NNDD ND FFSH463 NNDD 2200 o01l0e0s7 NNDD 2759195%4 NNDD FF441732 NNDD 2200 0ollml NNDD ND 7799995%89 NNDD ND Fs Fs22 NNDD 2200 00115s NNDD ND 5m04a20 NNDD ND DoseLeGvRelO:UP02mghg ~~ FFSsMW62k3l 0N0D6 2200 0018Bs 0N9D5 s0s22o0 7N7D FsMi2 F633 NNDD 2200 0olssl6 NNDD 032 8s13o7 NNDD 259 FFS4M4e2l NNDD 2200 001103 NNDD ND 629621657 NNDD ND EE3s44l3 NNDD 2200 o0n1s8 NNDD 6792236735 NNDD FF438832 NNDD 2200 o0u1s% NNDD ND nmxM NNDD ND Flsel F462 NNDD FFssuWs6LeL3 000022 FBsMusSi32 00002 Fs 004 2200 ooBnRe NNDD 2200 oollust 002390 2200 ooius? 00231 20 oun 07s HWHSTLSUM ND wB7m7 NNDD ND s7a8s1s7 221025 029 99445s1h3s4 32429 24 6787 519 03950 ()()()18p 2 SUMMARY of 6329-137 LIVER SAMPLES AMDT 22195.2 Dose GROUP3 Level: 125 mgkg ~ FSM4382 F534583 002 ND 20 20 0132 0101 026 ND 034 767873 198 767873 ND 259 FsM431 ND F532 ND 20 ou6l ND 20 om ND ND 764455 ND 764455 ND ND F534433 ND F51450-1 ND 20 0108 ND 20 013 ND 764455 768935 ND ND Fsus02 ND FS34503 ND 20 own ND 20 own ND ND 768935 ND 76895 ND ND FSMEL1 002 FsM672 ND 20 016s 023 20 0108 ND ND 7.5608 176 775604 ND ND F54673 ND 20 01004 ND 715604 ND F5u39-1 ND F592 ND 20 016 ND 20 om ND 817065 ND ND 817065 ND ND FFsSM343s913 ND ND 20 20 ous om ND ND 817065 95821 ND ND FF55i4445s32 ND ND 20 20 ooneml ND ND ND 99558812211 NNDD ND FF51s459}-1 ND ND 20 20 0opuB NNDD ND 778822441155 NNDD ND GROUP 4 FFsSiM4599-31 002 ND 20 20 onm omss 039 ND 78415 07962 303 ND DoselLevel: 250mghg FS34492 ND FS34493 ND 20 0ST ND 20 ous ND ND 7072 ND 0792 ND ND FSIS04-1 ND Fs15042 ND 20 opm ND 20 01368 ND 2342 ND ND m32 ND ND FSIS043 ND E5361 ND 20 oie ND 20 007 ND 823142 ND 770014 ND FFS5344556632 006 ND 20 20 O018 082 ons ND 027 7.1014 629 7.1014 ND 21 FF5suusse4-21 ND ND 20 20 01304 0114 ND ND ND 92.0439 ND 930439 ND ND Fs14543 ND 20 ons ND 92089 ND HWI37LSUM 03951 00010Pa1ge 2 3 Omnersr emowour:sosar Ls DAraes FiAcnaymip e +Cs ISKALs AnRAnTAIIWIDATALIVERSOt5741 $oaB Eom Hi iD Vm eTw 1d0m wiew oomm So SsSemmmaoan ooomsooaon wawx 57$0 SSSmmeaaaddtss ooonn omoonn lohoomm Swass o% on om BESTCOPY AVAILABLE WiBoo sSSemeaoas c1ie%o oitnno looommw WBWOeM uuvme ai0o Oven ao 2i0o oomse : oRbn[irroroaun ooaoonrr Si oun om o000 ooGmiumse ooomnm mmmeeesww mnswee Bo ome ow we Sn gGnh nFaoss oomor Wa3 pmsessnt oooon 0 doume oown wwamy ibwe 020 o0ss o01m0 msawme o8n% c 7 oErhx ewoTomnw am oaGoomr oe 0 ome ow mae we 3i 3 H3E vuveeRssk2 o00m0 o20 doem 0t0es Paget wn oo os oe wdN at rs nf3i uve: rman ooonnn 3 rman aor 01100 oaosmb ooonnw mnemae suoms wom em os ow wow Bo ope on lune mp X3xo rroo"aannhae ai0eo ww 0 1 aobmm oonn mmiems huey Bo Go raen aIaoo]r ban ao b0Br ooewn oosn mmaos neus Bom oe nom oe 8"5 roomuaers oaaoonr A oo oooenmme ooomim mnwauesn lsoaea Bo ons om mum un - b "bo rmi ramann 90 Ta we ooaooor im ew BBnooowman oooommh mBnoamww ii3wn STCOPY AVAILABLE 585 erawnme ooomomr 5 beens om 22B00 oooonsme ooeenwawmoanmn eiaem : FI5 0 awm ooommor ga wa on iBBooo ooomnmee ooinnn Bo oun S0m G0e0 oons oonn hnoow 8Sga5a a mreueaens "tw we aoaor0r nem 0n3oaowmn oooenm mwmaoammm mnTaei SZaa emeawnan ooowmm Ze man on B03o0 oootnwmee aionw obewuassn aoomxn 20 ma ow 3 Bo onm om mm om Prez Ww isms P02 Rf.un Tu we oow0m8 on ew 0Dooonm oowm moww oimm 3 eo - ss: 228 2cz8 ~ BEST COPY AVAILABLE Paes spr aaioson n os nan se nn Iores ]P1 [CoIs om3Beroamwe eiBomLoomm P]{ooomEmpmeemem pomme cocomeenoouooemm anoa mm oommS BESTCOPY AVAILABLE 3GToooommomsmmee LI memm oomhoowae i ouaoomm . bpossmpsnee: me o5i3 0 LBBBooooaoommmm 3mm oosooinxn oy asmsomawnmwoaaoounn &TSET mmee Jr, ww 552oo . i B BBoouoommsoetnmowammelnom 0 % s2En5r omem 0o5 f1Do imemmouoonn ommeuissEwaneu MSha e3pigs 5g rant al 5 imax 2B0 reFreeesnss uo Fer: o0aom0r 0% 12000 o0ommw ooonmw emTesssses nnonno 30 aw om ram on 3B7o 0 rrFeeeaerrr om aaoco0r asm is om B0o occmuieeo ooonwm ssTemsss 0esnus $G 0 F rSmes aaaooto Go fous ac 0wBo oooumm ooommw smseaammm wwennn COPY AVAILABLE GWao FFraeessss aooaor%r i Fass oo 33W00o 00oEummm ooownn mTTeeeaamss hnunww 3 ohm on mas son v"b eobewia oon ao0eo% Lio onaommese ooinmw as ie om wn om os on ew S5s50 saWe 8 Sa aaoc0i ao B0Loo oociimeees oaumn . wwwnnn Goommm ooossn ooonui womm g&5e wFFiieouneesr aaacoor Feauins ox 00Bo 00dmumm ooomwn roommee oamw 20 ows ow rome ie 1 FFFiaaetsosrcs a0ao%0 @ om as te am I200o0 oaonimee ooommw mmmoaaeee oosmme S2Sea6s Feoems: Soo Fees aooei% oor 200 ooomnne ooommn mrmiooonw aoiewus g Paez Wnsnzxs aw Ber sesss FI o2oo0wr 20 0I2o0 0oouminms ooomww wssooomm oo1mm7 0 one on nnnoewowsase Woobm aoaot0 0 Bo odchiammm oiimww im te om wwnn oamm ooss oonn mhse 5BneenaSse Boom iaatHoo 0noemww saa 1m a om ooomm mhiee o0%w oonm lmamw Wno ooSewaan aaoo 2 8 2 @ 2E2 S Pages 1995-05-03 08:38 Output of : 950502A1 Software : version 6.1 c1990,93 operator + bow Date of the Analysis : 1995-05-02 08:48 Analysis File Name : C:\SKALAR\DATA\HWIDATA\LIVERS\950502A1 CFlauloirbirdaetio1n.5order = Inverse Logarithm s1ope iss arene Result = 10l [x=-"5c=l] 5xcl == = Scclooorrprreeeccttofeeddthvveaalleuuleeecootffrottdhheee csoanmcpelentration aalz == a0 = -00..0000006050 -1.z0681 CFalluiobrriadteio1n order = 2 Correlation : r = 0.99926 Result =a2 * x* + al * x + a0 aaz== a0 = o0l.o0o0o0z0s0 -0l00641 Sampler Diluter NTuympbeer i+ iSAL000 WSaasmhple TTiimmee Ar Tine :: 15020 sseecc. 1. sec sTapkeeciaulp needle Height : | NSoinnegle 70 mm. dnieleudtlieonHeiFgahcttor dilution Volume ::: 281.005 mm mi. ReDsilauntpiloen runs :ii1 RUseeprrocfeisle :: No . mr PWe0g0ty% 323-37 1 03958 000105 I 1995-05-03 08:38 OutPut of : 950502A1 - Fluoride 1.5 SPiagtnhalnumtbyepre :: De3bubbled DesycsotleomrNumber :: 0Yes diReLsuatmeple :tNoo idiGl Touhtrpeusthold :: 04095 Window event : off Fluoride L ss12 ssTTaannddaarrdd :: IIggnnoorree 5543 ssTTaannddaarrdd :: IIggnnoorree 5s65 ssTTaannddaarrdd :: Ign0o.r1e50 5s87 sTsTaannddaarrdd :: 00..630000 55190 ssTTaannddaarrdd :: 11..250000 DOridmeernsi:onInv: ePrPsMe Logarithm tstrairgtgerValLuiemit :: 1580000 DSUec SPteAarkt sihgapneore :: P6o0in=teSdec MeeNadsureigwniornedow :: 17520 S4ec RFielgteenreration :: NNoo ofourtmuplaut:: #e.4as SPiagtnhalnumtbypeer :: De0bubbled sDeycsotleomrNumber :: N0o RdieLsuatmeple :: NNoo ddiilG oTuhtrpeusthold:: 40095 Window event : Off 03959 000109 ) 1995-05-03 08:38 OutPut of : 950502A1 s$l2 ssTTaannddaarrdd :: 00..003105 s43 ssTTaannddaarrdd :: 00..009600 556 sTsTaannddaarrdd :: 00..115200 $578 ssTTaannddaarrdd :: IIggnnoorree $5910 ssTTaannddaarrdd :: IIggnnoorree DOirmdeenrsi:on2 : PPK sttrairgtgerValLiumeit :: 1580000 SDeUc PSteAarkt shiagpneore i: P60ointeSdec eMNedasureigwnionrdeow :: 17520 3S% ec FRielgteenreration :i NNoo ofuotrpmuUtla :: c#4.:#=#c#33 33960 000110 [3 1995-05-03 08:38 OutPut of : 950502A1 Fluoride 1.5 Fluoride 1 Pon Pen Pos Typ Ident Ch Result ? Time Ch Result F Time 1 Wt iw 2a InitialTraWcaesrh Drift 33 3 101.405682 11481 21615 44 04..7u8a3n2e 386 4 0.7926 00 0 43 oswl 6 8532 StandarWdash1 Standard 2 333 00..007612 0.075 Standard 3 3 0.091 763286 44 0#..041s4e8s si0 4 0.0375 000 87 5s5 9 S6 Ssttaannddaarrdd 45 Standard 6 33 3 001111323 0.154 11206814 44 00..0093078 11641317 44 00..11428933 00 0 1101 5587 12 s9 sSttaannddaarr87dd 33 00.1621815 Standar9d 3 1.234 11976817 4400..2473058% 2135 40.6974 00 00 f1r3a 15 510 w standarDdrif1t0 Wash 33 3 11.540855 0.062 22438151 44 00..87082653 2721 4 #.4ees 00 0 1176 uuw 8u LLIIVVEERR BBLLKK 21 FS3442-1 33 3 00..00606A1 01075 23081385 3187 44 44..4#8e8n%s 4 0.0238 00 0 2109 uu 222 uu FF5533444422--32 33 00..006669 FF5S33446644--21 33 00..006656 33533660 4400..00003927 3708 40.0037 00 0 2238 uu 25 u FFS533445654--13 33 00..006675 F53455-2 3 00067 43086739 44 00..00002977 44421317 44 00..00005666 00 0 227 wa 28 u DWraisfht F53455-3 33 3 0101046723 0.063 44851827 44 #0..e7e9n1s5 4925 40.0109 00 00 3209 uu 31 ou LILVIEVRERBBLLKK 21 33 00..007646 LIVER SEK 1 3 01105 55219019 44 00..00202344 5463 4 0.0798 32 u LIVER SPK 2 3 0.103 00 0 33 35 u FF5S33444411--21 33 000007772 F53441-3 3 01070 s5e6l4s1 44 00..00277746 65196877 440.0.00113604 00 00 3376 uuw EL FFS533445511--21 33 00l.o0s6s8 Drift 3 11502 66531355 44 00..00007780 6683 4 0.8013 00 0 i39 uWw au F5345W1a-s3h F53461-1 333 00..006692 0.069 76092297 440.#.0s1#0e6 7214 40.0114 00 0 4432 ouu 44 ou FFS533446611--32 33 00..006677 FS3460-1 3 01066 77358663 4400..00006511 7740 40.0044 00 0 4455 ouu 47 u F5533446600--32 F53447-1 333 i0/006665 0.065 07980s8 44 00..00101314 8262 4 0.0037 00 0 4458 ouu 50 a FF5533444477--23 33 001.006677 Drift 3 1517 88641460 750 4400..00005486 4 0.8078 00 0 51 ow 5532 wuu FS345W2a-s1h F53452-2 3 33 0.062 00l.00s668 9029 99311345 4 #.eens 440.0.00003708 0 oo Page 1 of 2 039s61 000111 13 1995-05-03 08:38 OutPut of : 950502A1 Fluoride 1.5 Fluoride L Pos Typ 5554 uu 5576 uu 5598 uu 6610 uu 6632 wa 6654 uu 6676 uu 6698 uu 770 oaw WE rw Ident F534B5LK2-31 SBLPKK12 FS34S6P3K-12 FF5S33446633--32 DWraisfht FFS533444444--21 FF5533444484--13 FF5533444488--32 DWraisfht RunOut Wash PEM Ch Result 33 00..006686 33 00..100676 33 00..019140 33 00..006770 33 01..056129 33 00..006821 33 00..006668 33 00..006664 33 10..056321 3 0.062 F Time 99646815 109081369 1100316971 1100751357 1110182869 1111422359 1111756953 1121914118 1122522991 12766 PEM Ch Result F Time 4400..00007389 o0 4400..00082354 00 440.0.00681837 00 4400..00015185 00 4404..5#0#8e5s 00 440#..0#s3e6e3k 00 4400..00007352 00 44 #0..#0a0r3s9 0o 44 0#..#8#1r3s5 00 4 #.#e84 Page 2 of 2 03962 00011: " Fr 1i0.2097253 mo| J Po:Lo]j |HH :| || { |i | 1 | || Calibration curve of s50N0EAY Flooriset Ehi Ssod A/ )Hi So s 1 / || ye I A/ || ,/ | / 1 J I A l / 1[| | |ilo006409SorE der + 2 -_-- rm - || | ross | riomms 3963 000117 1S i| | 11.818152{8 oNI 7 1 } | Lfoo | g I[1I os" [od | | lo| J| o | tion cre af ston erin 15 AH A | :I / i / S/S / / Jo) / / i / {l / | SF I /// (iBI I [I | 1| -- = || 03954 000113 1% I | Coog l. i } C1 | Co ary re sn | I oo |I] Poi PI |AHh Hi H! CH Fo oH UVaaaUU P|o HHE HHH ahitIH HT sh HT|H 3A [ Vit Vou | 03965 000115 nn foto ot sO rie 5 || 4055 | ETT re -- | H| | on H | wl od iil |] |If | | I I EE | | lo] HHi H HHH CoLaL |1 Posh i CITI, | [oy Se aAREA Vm | SERRETEE] 03966 00011. 18 tof rrr 5 | 4035 Po i vo] 2% i | | Lo | |Po | i PCo d |! ly] | Lil I { bo 1 I[ Po ll Cr Po i 1 (1 || !| i} 3nha2 | PAAJ bod I ses | RANT JNA Tine 1| I |i i [Ii i! II I 1i IH I iI H| ALA J 662|3 03987 000117 Ia} Rou data of 905001 Fluoride 15 ET a *| CoE| Tav . |! TH1 if 1 i Pool ill [ Poa I Iiah E no 3! ! ii| ii| | i Ji1 i I iI II i!i i| Co i| I Hli [i II I } || iI II IIB i[i J|AfR e 9NwAaRg to] BRus ED Eyr N og se a> ~ | we | Tine i wis| ESET 1Fils ora pe C3958 00011% 20 os wos (ICHof Me-- i I 1 " HIi i | pe 1 | 1 i I i i I i| {i I i Ht | | i {:4| 1 i IIh|| HI I | l | i I I i|| a[Il) [IHil ] |1! II anes Rr 2 x (Rew UE ed Wt ey Ne wall A| 0 TT TT1 Seba 1 Fie | RE pT 03969 000119 PAL - EE ET [REY SETH ied BOY 510 Eyre say as {ol | 1 ItI I |loll {] I i| 1Po || || ll | I & a Naa * ds -- aI |fH | i i h|] | I IH | | | I| i| ! ol 003970 000120 ne 1995-05-03 09:05 ouerue of : as0s0z81 voxator Software : bate of the version Anslreis oo 6.1 1990,93 1985-05-02 12:41 EE Prortais 1.5 Stove ca suerene roma = 30[Fo2a)] dxsricagorprmeceatednvamteioef tghe mseemeame F2aE:s 0.0000 3 EDorEiaE onaer = 2 a Corsetation : 5 = 0.35532 2FBr oooomm soto ME1ene ii: lfsontonseoe. MA FiOhoENnE | 1"m,e ET dBeigitnes Yo 2 va rie Coen pug#9 O0w "he see 03971 000121 3 1995-05-03 09:05 output of ; 95050281 Fluoride 1.5 Errlatorhnarnlu'mTboeer | po3busbiea EaReylseiavvmeepnienumber a Thresnosa i|Choo | 100s ndaioswupvaene | B8er 2s2 fsihtaaannndddeearrrdad Canard |:| | di1oggnnnooorrrsee doer 575 fo SrStTaaannndddeaerrrddd ||| lem5bon7is0a ihandera | olees CHodr]afunnTddaeveerdsss|| Sinansion: Bou Log1Dtii0gosh FSEorarirkotseohrvaariet'nsi Src ionere |||: B51o00t000ntsbSaeuc | 50S HawoissureSnwoirnedow [| fh7%5e0 4e tRfaaugemtnearatioLnann| eho Fluoride L DpSeaoctnahallen:u"TmTPebeCer:I| oRda0subsied dRSeyisevacemespnleumber | | RN0oo HdiTnsdoooutvnearnot ||| B80e0es 03972 0001222 Poged&s 1995-05-03 09:05 OutPut of : 95050281 5521 ssTTaannddaarrdd :: 00..003105 s543 ssTTaannddaarrdd :: 00..009600 $$65 ssTTaannddaarrdd :: 00..115200 $s78 ssTTaannddaarrdd :: IIggnnoorree 55910 ssTTaannddaarrdd :: IIggnnoorree DOridmeernsi:on2 : PPM tstrairgtgerVaLliumeit :: 1580000 SDeUc SPteAarkt sihganpoere ii P6o0inteSdec MeeNadsureigwnionredow :: 71520 Sec RFielgteenreration :: NNoo ofourtmUplaut:: 4c.44:8=8c%3 03973 000123 24 1995-05-03 09:05 Output of : 95050281 Fluoride 1.5 Pluoride L Pry Pom Pos Typ Ident Ch Result F Time Ch Result F Time W1 t iw 2a InitialTraWcaesrh Drift 33 3 10..408576 1.491 21615 44 #0..a8n5n0y6 387 4 0.e536 00 0 43 sowl 5 52 standarWdash1 33 o00.l005674 Standard 2 3 0.073 673278 44 #0..40a1n43# 913 4 0.0324 00 0 7 5s43 8 s5 SSttaannddaarrdd 43 33 00.1100877 Standard 5 3 01131 11206849 44 00..00859766 1440 4 0.1221 000 19,0 s567 11 58 SSttaannddaarrdd 76 33 00..218564 Standard 8 3 0603 111967618438 444 000...412546497103 00 0 1132 14 5910 a StSatnadnadradrd109 Drift 33 3 11.1326375 1.514 22311328 44 00..87430070 2488 4 0.8862 00 0 165 owu 17 u FS344W6a-s1h 33 00..006547 F53446-2 3 0.062 22872293 44 0#..0#1a3e0k 3004 4 0.0100 00 0 1198 uu 20 u FF5S33445436--13 33 00..006656 F53453-2 3 0.065 33319610 4400..00117722 3539 4 0.0158 00 0 2221 uu 23 u FF5533445583--13 33 00..007719 F53458-2 3 0.066 33879113 4400..00422779 4065 40.0172 000 2258 wuu 2a FF5533444538--13 33 001.006630 Drift 3 1.476 44420357 4400..00101576 4587 40.8457 00 0 2278 wu 29 u F5344W3a-s2h 33 00.006587 F53443-3 3 0.062 44984216 440.#.0#2a2e8n 5113 40.0079 000 3310 ouu 32 u FFS533445500--12 33 00..006628 53450-3 3 0.060 55426839 4400..00120103 5635 40.0047 000 3334 ouu 35 ou FF5533446677--12 33 00..006650 F53467-3 3 0.060 55891877 4400..00105453 6162 40.0045 000 3EY% u 38 a FF5533443399--21 33 00.006602 Drift 3 11515 66353176 4400..00004932 6688 40.8665 00 0 309 ouw 41 ou F534W39a-s3h 33 00..007537 FS3445-1 3 0.064 76053287 44#0..#0e3e2w6 7214 40.0136 00 0 4432 ouu 44 ou FFS533444455--32 33 00..006620 F53459-1 3 0063 77336844 440.0.00009461 7738 40.0109 00 0 4465 uu 47 u $F5533445599--32 33 00.006680 BLK 2 3 0.068 87095108 4400..00202367 8266 4 0.0215 00 0 4485 uu 5510 wa SBLPKK13 33 00..009515 Drift 3 1.483 88641366 8790 44 04..046a4808 4 0.8452 00 0 Wash 3 0.057 8930 E52 au SSPPKK'23 33 001.00899 9141 44 0#..0e6e0e3s 9317 40.0688 00 Page 1 of 2 03974 000129 5 1995-05-03 09:05 output of: 95050281 Page 2 of 2 Fluoride 1.5 Fluoride Pom Pon Pos Typ Ident Ch Result F Time Ch Result F Time $554 uu 56 ou L5IV3E4RS4PKB9L1K4 33 3 00..015090 01061 694353 9841 44400..0.000070783568 57 u FS3449-2 3 01036 10013 44.4883 000 5598 6 ouu u FFF55S333545040454--123 33 3 001005850 01054 1100316819 10535 44440.#.l08e0p84e47n 00 00 66&12 4u 65 wu FS3D5r0i4f-t3 Wash 333 011.045579 0.057 11l10o07s41e41s 440.#8le3e6e6s 4 4.#88s F53456-1 3 01038 11237 40.0007 000 e666 uuu 5PP3SS433535664--132 33 3 00011000558717 1111543133 11763 4440.0#l.0e00e57e79s0 00 00 86 70 uu u nou PP5S33B44L55K44-231 33 3 000.11000565807 111127512308388 4440.04.0.0#00e05s5%s BLK 2 3 0.057 12463 4 4.6883 0 0 7732 uou 74 a SSEPKK21 Drift 333 00l1111013 11498 1122861471 12951 44 00..00893549 40.0530 00 00 7765 u 77"8 oau SWPaKs'h3 SPK 4 333 001015875 01164 111333523124725 4440.40.1453l9090lald00osu cuosohs Drift 3 1l518 13esi 4 0.gsac w7t wrw Runout VVaasshh 33 00.005577 1143186960 44 ##..s9e8e8sh 00 03975 000125 % To. ss0ms | ii P | o| || 1| || g1 2 || SoA / JS | J"4 Ii 'JS I| l| | i / i [glil // || py / /I I | ] J Hfll | / | lL||| 7{ iI L |p{0 onoim ordero : 2E wn r: e-- 0.smem I|| C03976 000126 27 Strato eo sn pri 5 | Ri PA i |lo | | { i L IH g |i / I i yal /7 |0| /J {| // [i |} / fH | I / |iy lo|| 1| // 1 {I|| |1 I AI || ||L0 o Ord'er In' vem rse LegarithnMe easured i 5: ' osn mm 2085 || 03977 000127 2% Co i || ey 40s 1E TryT re meen i | E| E. ar|I [I | Cor Ii h TH Fol PE | | [I I { { hi Cl min FoLa | RmE n L1ol H |Pod Li E VII I wh HH MH fritid A CL a AY | AUN YY 01 | 03978 000125 ; a TI I pod 2 C'o [I | : nol Pl Po hh H un i | | i i lod o| | [ CH HH 1| Coe || | 4 f | |Pi | Lr hh |[1||1 RI AA TATRIAE T { = CUT mm2a T| idm . := 03979 000129 5 | || T e o ee.| Ce | | 1 iy | | | ||| ]| || .l A|i [| 5] | CL | | 1 | i || || |I || ll| i || I[ Hi p[= ot i S_--ARFAR Am II Vr 03980 , 00015:0 Cl i. | 2oes rT ; AA T | {|| | 1h ![ || JIl {IIl |i | {| | | | || Ii |I FIo ol 1 I i Ll I] LC|o lif 2[|o1o sI |Dl a AAmAaImmNaINNAYNaMdl] | -- Tw | 03981 000131 = [Rau dta of OSB Fluoride 16 -- ass S eee E I | bod | 1 I i { | Cd !1 i| il| I ||i I|l I| Iih Il | 4] I [II os lL] | Io i |!| |i I|| | ]| 1i || i I 23ot |Ii P { HA E lg se<E 4as 8$1R ev[ ) \ewla o [Ile | " Tie EsceExit | FieHeln | Crtl-Peait pears | | iIn | |! ibi | Hl |]| iI i1il || I [ Reee l)|i Tni 03982 000132 | cos aices_iive KIER aloo:EEN vein= LIo | I I i I i J BE | I | atI i HH | I | I } | | | l ' | | | | | bo sl Ahaad =a {Hl [ | deOtoRM AUMGUY Na [l | Er a 03983 0001533 34