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3E M nvir Laboo raton ry me ~~ ntal Final Report- Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T6052 in Rabbits In-Vivo Study Reference Number: HWI#6329-135 TSetsutdySuNbusmtbanecre:: AFMCD-1T2-002(0T7-69055.21) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facility: 3M Environmental Technology & Services 935 Bush Avenue St. Paul, MN 55106 Method Numbers AMDT-M-1-0, and Revisions: Thermal Extraction of Fluoride by Means ofa Modified AMDT-M-2-0, FDlouhorrimdaenMneDaXsu2r0e0m0enOtrgbayniMceaHanlsiodfeaAnnaOlryizoenr-ELAiv9e4r0 Expandable Ton Analyzer AAMMDDTT--MM--54--00,, AExntarlaycstiisonofoRfaFblbuiotrLoicvheermiEcxatlrsacftrfoomr RFalbuboirtocLhievmeircals Using AMDT-M-8-0, AElneacltyrsoissporafyFlMuaosrsidSepUecstirnogmtehterySkalar Segmented Flow Analyzer AMDT-M-14-0, with Ton Thermal Selective Electrode Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum Initiation Date: See attached protocol Author: James D. Johnson Approved By: arn tudys DDi.reJdoiheftisbn 00630 ze [2c Completion Date --_ h-o Samples of `combustion lfiovrertoattal28ordgaaynsicpofsltuodrienrem.alEvdoesnesthoefhFiCgh-e1s2t0do(sTe-6o0f521)00w0ermeg/ankaglyzed by (tr5a0ceuogf/ksg)omreesfullutoerdiinne ndoetoercgtaabnliec ifflouonreinueseast jpursatctrieclaaltiqvueanmteitteartiroenadlienveglss.. There is a Electrospray mass spectrometry perfluorodecanesulfonate anion. was able to detect m/z=599 ion which is the `The doses were too low to assess dermal absorption with this test method. 20INTRODUCTION ~~~ 000000 `5T0heugp/hkagr.maTchouksi,neitnitchisstustduydfyo,riFfCt-he1r2e0wwaassansoutbssutacncteisaslfual.moTunhte ohifgohregsatndiocsfeluwoarsine present at occurred. 28 days it However, would in the indicate event of tvheartyalistitglneiofricgaannticabfslouroprtiineonaotf28FCd-a1ys2,0 had it would not be serum possible samples wtoermeakaveaialnabalsesfeosrsmaenanltyosifsdoefrtmoatlalabosrograpntiiconf.luLoriivneer samples and and perfluorodecanesulfonate anion. quite low (high dose 50 ug/kg), The samples were it was not expected analyzed. Because the that fluorine would be doses are detected. 3T 0 ESTMAT ~E ~R ~ IALS 3.1 Test, Control, and 3.1.1 Analytical Reference Reference Substances Substance: and Matrices FC-95, lot 161 or 171. They are equivalent. 3.1.2 3.1.3 Analytical Analytical RCoenfterroelncSeuMbasttarnicxe::BNovoinnee liver, bovine serum, rabbit serum 3.1.4 Analytical serum Control Matrix: Bovine liver, bovine serum and rabbit 3g.r2ocSeoruyrscteoroe,fbMoavtienreiaslesr:u3mMfrIoCmPS/iPgCmPaDCihviesmiiocnalfoCroFmCp-a95n,y,borvaibbnietlsieverrufmrformom AMDT-110394.1 (HWI#6329-123) control group animals. 3.3. Purity and Strength of Reference Substance: Responsibility of Sponsor. 3.4 Stability of Reference Substance: To be determined by Sponsor. 00631 -- = 3bi.o5lSogtiocraalgseaCmopnldeisttihoensstofroargeTeisst-2M0a+t1er0iaCl.s: Room temperature for FC-95, For 3R.e6guDliastpioosnitfioornthSepetcimiemepnesr:iodBiroelqougiirceadlftoirsssuteusdiaensdloflnugiedrs twhialnl b2e8 rdeatyasi.ned per GLP 4.0 EXPERIMENTAL - Overview aTnhaelytsiisssufeosr ffrluoomrianneimcaolmspoduonsdesd.asAdtestchreidbiesdcr(eHtWi1o#n6o3f2t9h-e1s3t5u)d,y wdeirreectaovra,ilaasbelreiefsorof acnoamlbyutisctailotneswtasscotuheldmobsetpelirkfeolrymaenda.lyTsihsetsocprreeesneinntgdfeofrinfiltuiovreiddeatian floirvearbvsioarption if the pharmacokinetic Other available tests wteesrte(eIlVeacdtmrionsipsrtaryamtaiosns) was positive spectroscopy. for fluorine However, in if the the liver, sdeafmipnlietisveworuesludltbsecoaunladlybzeedobatnadinaendywoitthhercotemsbtusswtoiuolndanbaelyfsoirscaolnofnier,maotniloyn.the liver 5E .0 XPERIMENTAL-METH ~~ODS D5.o1hrAmMaDnTn-MD-X12-000,0 Thermal Organic HEaxltriadcetAionnaloyfzeFrl-uoLriivdeer by Means ofa Modified 5.2 AMDT-M-2-0, Fluoride Expandable Ton Analyzer Measurement by Means of an Orion EA940 5.3 AMDT-M~4-0, Extraction of Fluorochemicals from Rabbit Liver E5l.e4cAtrMoDsTpr-aMy-M5a-s0s,SApneacltyrsoimsetorfyRabbit Liver Extract for Fluorochemicals Using w5i.5thAIMoDnTS-eMle-c8t-iv0e,ElAencatlryosdies of Fluoride Using the Skalar Segmented Flow Analyzer D5.o6hArMmaDnTn-MD-X1240-000,OTrhgearnmicalHaElxitdreacAtniaonlyozfeFrl-uSoerriudme by Means ofaModified 06632 n=e 60DATAANALYSIS~~~ The data from combustion analysis are attached. There does not appear to be any stoatmaplloersgaanti4c8fhluoourrisnepoasbtoivnetrtahevepnroaucstidcoaslequfaonrtaitnaytoifotnhleimditosfionrganreygoimfetnhse. liver Even the 50 ug/kg dose is not above the practical quantitation limit. However, if one uses just the meter readings and compares those reading to the values for the controls, there is possibly a trace of fluorine being detected. Electrospray mass spectrometry analysis is attached. The electrospray was able to detect perfluorodecanesulfonate `present was not quantitated. anion in the 50 ug/kg dose livers. The amount In viewofthe failure of the pharmacokinetic study (HWI#6329-134) to show a good `marker for FC-120 at a dose of 50 ug/kg, it is not reasonable to make an assessment of the extent of dermal absorption from this study at the same dose level. Other data was collected using Skalar segmented flow analyzer with ion selective electrode (see appendices). This data, although supportive, in the opinion of the Study Director is not required to reach the conclusion stated here and therefore is not discussed in detail. p6.r1obCliermcuwmisthtatnhicsesanatlhyastisMiasythHatatvhee ApfhfaercmtaecdoktihneetQiucalsittudyyodfitdhneoDtaptrao:vidTehae good marker for dermal absorption because the dose level was too low. The dermal study is not at a higher dose. Zo coNcrysioN 00000 `This study does not provide a useful assessment of dermal absorption of FC-120. There is not a useful marker. 8.0 MAINTENANCE OF RAW DATA AND RECORDS 8.1 Raw Data and Data: Raw data, approved protocol, approved final report, appropriate specimens, and electronic data will be maintained in the AMDT archives. 00533 ---- = 90APPENDICES 9.1 Protocol and Amendments 9.1.1 Protocol and Final Report: HWI#6329-135: "Single-Dose Dermal Absorption/Toxicity Study of T-6052 in Rabbits" (Protocol type `TP3016.AB for dosing of animals, tissue collection, etc.) 9.1.2 Analytical protocol AMDT-020795.1 9.1.3 Amendment to Analytical Protocol AMDT-020795.1 9.2 Signed Reports from Individual Scientists: None 9.3 Quality Assurance Unit Statement: See attached 9.4 Key Personnel Involved in the Study: See attached 9.5 Materials and Equipment: See methods 9.6 Solutions, Reagents, and Standards: See methods 9.7 Sample Preparation: See methods 9.8 Quality Control Practices: See methods 9.9 Test Methods: See Protocol AMDT-020795.1 9.10 Instrument Settings: See methods 9.11 Data: See attached. 9.11.1 Summary and raw data; ug F" in whole liver as determined by thermal extraction followed by analysis using Orion ion analyzer. 9.11.2 Summary and raw data; analysisofliver extracts using electrospray mass spectrometry. 06634 = 9.11.3 Summary and raw data; ppm Fin serum as determined by thermal extraction followed by analysis using Orion ion analyzer. 9.11.4 Summary and raw data; ppm F" in serum as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. 00635 az 4, 9.1.1 Protocol and Final Report: HWI#6329-135: "Single-Dose Dermal Absorption/Toxicity Study of T-6052 in Rabbits" (Protocol type TP3016.AB for dosing of animals, tissue collection, etc.) 00636 me WISCONSIN TRIS BIAT 800 5808 Sponsor: 3M ToxicoloSgty. SPearuvli,ceMiMnendeiscoatla Department +CORNING Coan EER FINAL REPORT Study Title: i{: suit 22 19 \o, 4 3 "Jicors Single-SDtousdey DoefrmTa-l605A2bsoirnptRaibobni/tTsoxicity Author: Steven M. Glaza Study Completion Date: June 27, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard : Madison, Wisconsin 53704 LabPorojrectaIdetntioficratiyon: WI 6329-135 Phone Gos 241 ae71 Page 1 of 41 fax Cco00537. __=-. OPTUS MAIL DUUVIRY 2001 LINSMAN RIVE MADISON. Wo aerwn Page 2 of 41 HWI 6329-135 QUALITY ASSURANCE STATEMENT TW(hiFisORsc)onrseGiopnoo,drtLIahnacb.so,rabteioenrnyacrcePovrriaedcwatenidccee bwRyietgthuhleatthQieuoanlFsio,toyd21AansdsCuFRrDarnu5cg8e.3A5Udnmii(tbn)isotf(Hr6)aatzi7lo)en.ton The following inspections were Study Director and management. conducted and findings reported to the Written status reports of `inspections asntdandfairnddinogpseraatriengispsruoedcedtuoreHsa.zleton management monthly according to Inspection Dates From To Phase Date Reported to StudyDirector Date to Management . 12/21/94 12/21/94 Protocol Review 01/30/95 01/30/95 Protocol Amendment 12/22/94 01/30/95 01/10/95 02/10/95 02/02/95 02/02/95 Body Weight 03/29/95 03/29/95 Data/Report Review 02/02/95 03/29/95 03/10/95 04/10/95 03/29/95 03/29/95 Data Review 03/29/95 04/10/95 06/27/95 06/27/95 Report Rereview 06/27/95 07/10/95 A Representative, Quality Assurance Unit 27.95 Date 00538 = Page 3 of 41 STUDY IDENTIFICATION single-SDtousdey DoefrmTa-l605A2bsoinrpRtaibobni/tTsoxicity HHI 6329-135 Test Material T-6052 . Sponsor 3M ToMxeidcioclaolgyDeSpearvritcmeent 3P.M0.CenBtoexr,332B2ld0g. 220-2-02 St. Paul, MN 55133-3220 Sponsor's Representative J3oMhn L. Butenhoff, PhD ToMxeidcioclaolgyDeSpearvritcmeent 3PM.0.CenBtoexr,332B2l0dg. 220-2-02 S(t6.12)Pau7l3,3-1M9N62 55133-3220 Study Director ' HSatzelveetnonM.WiGslcaoznasin, Inc. P.0. Box 7545 M(a6d08i)son2,41W-I729253707-7545 Study Location BHuaizlldeitnogn WNoi.sco3nsin, Inc. M3a8d0i2soPna,ckeWIrs A5v3e70n4ue StSutduydyTimIentiatbilaetion Date Decenber 30, 1994 Experimental (In-1ife) In-ife End Date Start Date JFaenburaurayry5,2, 19199595 : SEtxupderyimCeonmtpalletiToenrmiDnaatetion Date JJuunnee 2277,, 11999955 00539 net Page 4 of 41 Acute Toxicology Steven M. Glaza Study Director Manager SFtruadncyisCoo(rBuddi)naWt.orMcDonald Patricia Padgham In-Tife Supervisor Rose M. Bridge Report Supervisor Quality Assurance Sherry R. W. Petsel Manager HWI 6329-135 KEY PERSONNEL LaboratoryAninalMedicine Cindy J. Cary, DVM Diplomate, ACLAM Supervisor Anatonical Patholoay Thomas E. Palmer, PhD Anatomical Pathologist Jack Serfort/ DSNeuebpcoerrroavphissyoL.rs Pirkel Anne Mosher Supervisor Pathology Data COC530 -- Page 5 of 41 CONTENTS QStuuadlyityIdAesnstuirfaicnacteioSntatement Key Personnel SOubmjmeacrtyive TReesgtulaatnodryConCtormoplliaMnacteerials PrToesctedSuyrsetsen DRiesscuulstssion SRiegfnearteunrcee Pathology Report Table 21 IInnddiivviidduuaall aClnidniMceaaln BSiogdnysWeights (g) 34 IInnddiivviidduuaall DPeartmhaollogIyrrCiotmamteinotnsScores 5 Individual Animal Tissue Weights and Bile Volumes Appendix A PPrroottooccoo]l ATmPe3n0d1m6e.nAtB No. 1 HHI 6329-135 Page 32 . 2 86 88 109 1133 114 15 1176 2139 25 27 28 20 00541 ---- Page 6 of 41 HHI 6329-135 SUMMARY sTkhiins sitrurdiytanwcays odfonTe-6t0o52aswsheesns atphepliseydstetomictheabsskoirnptioofn/ratbobxiitcsi.ty and relative TohfethsetuHdrya:wa(sNZWc)oSnPdFuctsetdraiunsinfgortheraeceh mtraelaetmaenndttghrroeuep.female acclimated rabbits Group Test Material Do_(smea/kLae)vel MNalu es m ofbAFnee immaalrless 1 (Con2trol) DiTs-ti6l0l5e2d water 2o 33 33 34 T-T-66005522 1,020000 33 33 a Adninistered at a dose volume of 2.0 mL/kg. Trheespbeacctkiveofmaetaecrhiarlababtit twheas incdliicppaetded fdroesee olfevhealirwaasndadanisniinsgtleereddoseto otfhetheskin woafstchoeverraebdbitwsi.th Tahgeautzreeatbmaenndtagesitseescurreedmawiintehd pianptearct.tapeThearoaurneda oalfl aepdpgleiscatainodn dorveesrswirnagppefdorwiath24S-ahroaunr HexrpaopsuraendpeErliaosdt.oplast tape to provide an occlusive , C4lihnoiucrasl afotbesrervtaetstionosr wcoenrteroclondmautcetreidalpreaddomsienisatnrdataitona.pprAodxdiimtaitoenlayl 1,1i2n.i5,caland ofobrse2r8vatdaiyosn.s aBndodytwwiecieghatsdawyermeordteatleirtmyincehdeckosn wDearye-8confodructreadnddoamiizlaytitohnereafter piunr-ploisfees,terbmeifnoarteiotnest(Doary c29o)n.trolThematienriitailal addemrimnailstriartriiotnati(oDnayre1a),dinagndwaast made breeafdoirneg)t.estSuorbsecqounternotlremaatdeirnigaslofaddmeirnmiasltraitrirointat(iroencowredreed maasdetheapDparyo-x1imately w3e0remicnoultleesctaefdterfrobmandaamgaergrienmaolvalear(Dvayein2)ofantdheonanDiamyasls4 baenfdor8e. inB-l1oo1dfesamples ainnditi22a.tion"In (aDdadyit1i)o,n,appatrotxhiemattienley o2f4-hnoeucrrsopspyostondosDeay(2D9a,y a2)p,proonximDaatyesly4, 208,'mL15, toseferpmbailrnoaaottdeidownaisntooofbttshaeeirnuemidn-alnfdirfoemcelephlaacushlearanainfmdraalnc.teicornoAspl.lsiesdAa.lmlplTeahsneimwwaehlroselewceernleitvreeiruf,tuhgbaeindlieza,enddanat aanpipmraolxsi,mataend 1b-octmhxki1d-ncenyssecftrionononoef mtahleedearnmdaloneappfelmiaclaetioinn seiatceh fgrroomupalwlere cbloololdectseadmplaetsne(csreorpusmyanadndcweelilguhleadr f(rvaocltuimoenso)n,lylidveetresr,minbeidle,fodrerbmialle).applThiecation siint-e1si,feapnhdaskei.dneys were sent frozen to the Sponsor after termination of the _ 00542 -- Page 7 of 41 HHI 6329-135 bAopdpyliwceaitgihotn goafinT-6o0r52macdriodscnoopticresfuilntdinignsanaty clinically normal throughout the study. No tneecsrtopmsayt.eriaAlll-realnaitneadlschaapnpgeeasredin dermal irritation was observed at otrheT-d6er0m5a2l atscoarniyngofitnhteerdvaolsse alsevealsr.esult of the application of distilled water 06643 Rn Page 8 of 41 OBJECTIVE HUI 6329-135 rTheelatoibvjeectsikvien oifrrtihtiasncystoudfyawatsesttomaatsesreisasl twhheensyasptpelmiicedtotoxitchietys/kaibnsoorfption and rabbits. REGULATORY COMPLIANCE ATdhmiisnisstturdaytiwoans'sconGdooudcteLdaboinrataocrcyordParnacceticweithRegthuelatUi.So.ns Fofoodr and Drug Nonclinical mLaatbeorriaatlormyixSttuurdieesp,rep21areCdFRfo58r, twhiethGrotuhpe e2xcaenpitmiaolns tfohratcoanncaelnytsriastioofn,the test hinomotghiesneisttyu/dsyolaurbeiliintyc,ompalnidanscteabwiiltihtytwhaesAnniomtalconWdeulcftaerde.ActAllRegpurloacteidounrse.s usIend tdhuepliocpaitneionanyofprtehveioSupsonwsoorrk.and study director, the study did not unnecessarily TEST AND CONTROL MATERIALS . Identification TThiequitde.st mThaetercioanltrowlasmaitdeernitailfiweadsasdisTt-i6l05l2edawnadtdeerscarnidbewdasasdesacrcilebaerd, ascolaorless clear, colorless 1iquid. Puar ndi Stat biliyty TdheeterSmpionnastoironasssu(miensclurdeisnpgonsuinbdierlittyestfocrontdeisttiomnast)e.rialAnapluyrsiitsy oafndthsetabteisltity . mhaotmeorgieanleitmyi/xstoulruebiplrietpya,redandforstathbeiliGtryouwpas2 animals for concentration, not conducted or requested by the Sbeponasdoerq.uateTheforpurtihteypuarnpdossetsaboiflittyhisofsttuhdey.control material were considered to Stoanr d Ra eteg ntie on Tshaemplteestofaendachconttersotl amndatecronitarloslwemarteersitaolredwasattarkoeonm atnedmpweirlalturbee. reAtairneesdervien a wfirtehezeHrazlseettontoWimsacionntsaiinn a(HWtIe)mpSetraantduarredoOfpe-r2a0tiCng10Profcoerdur1e0 y(eSaOrP)s. inAnayccournduasnecde tpehsatsemaatcecroiradlingwastorHeNtIurnSeOPd. toAntyhereSmpaoinnsionrgafcotnetrroclommpalteetriioanl ofisthreetaiinn-e1difefor other testing and will not be discarded after issuance of the final report. C0644 Page 9 of 41 Safety Precautions HHI 6329-135 Tahned ptoelsitciaensd. contro) material handling procedures were according to HHI SOPs Test Aninal TEST SYSTEM . AWDidesunvcleotrn,sailnPbeinfnnaoscyirllavibatbnyiitasatoo3nf80Dt2ehecePmaHrbcake:err(s2NZ8AW,v)1eS9nP9uFe4,satMnradadiimnsaoiwnne,traeiWnipesrdcoocnuastriendt.hefroHamzlHRePt,on Inc., ; Housing ADfutrienrgreaccecilpitm,atitohne aanndimatlhsrouwgehroeutactchleimasttueddy,fotrhea housed in screen-botton stainless steel cages in apneirmiaolds owferaet temperature- ilenadsitvid7uadlaylsy. and humidity- mc1a2oi-nnhttorauoirlnleTdia ghqttue/am1rp2tee-rrhasot.uurredEaonrfvkir19oTnimgehtntotian2lg3Ccc,yocnltaer.orleslaIntfiocvraestehhseumwiahdneiirtmeayl ovafrroio5am0t%iwoen2rs0e%,Fsreotamndto a tahdevseerseconefdfietcitonsonextihsetesdt,udytheoyutcwoemree. documented and considered to have hag no imal oi TLhaeboraantionraylsRawbebrietprDioevtidHeFd a#5c3c2e6s,s analyzed by the manufacturer for tPHoIwaFteeedrs,adIn1ci.bitTuhmeanfdeeda miesasruoruetdinealmyount nutritional components and environmental of . wcienortneetrafmneoirneakdnntosww.inthcoSnaomtrpalmaeifsnfaencottfsedthitnehewthaetreesrfueleadtrseoropfewrtaihtoeedrisctaautldlyyl.evaenlaslyztehdatbwyouHlHId. havTehere Selection of Test Animals wpTrehoregeraanpmli.amcaeldsThewienrrtoaendisotdmueidnzytaitfgiirooenudpsbboyduysainniwgmeailgahtnssturmwabeterirefiadeneddtecbroomdriyrneeswdepiognonhdtinDagryanead-8ro.mitzaagTthieaonnd Wbsoetdiuygdhytwediivgdahrtinsoattifooenrxceoeeafcdht4h2geroasuntpiamnadolafsrdeafcdoherviseaeaxtcihownegsrreouonpfotothfestemaaectahinstsiwecexiaglshletyl,edcitafendfdertfehonertmtehaaten atsnhtoeutdhy5e%rprpirfoeormbaalbteoilit(trNyoe.atlmFee5vn2et9l.82d)ue"Ownhteiocfhpeomwoaarlsehetaarnleitmahat.led (TNihon.isthFe5a3ni4sm0aa9ml)e wwaass rreeppllaacceedd manner. wiinththe - 06545 a Page 10 of 41 HHT 6329-135 StudyDesign Ainnitmoaltshewefoilglhoiwnigngfrosntud2y,05g2routpos:2,471 g at initiation of treatment were placed Group TestMaterial Dose Level _(ma/kq) N Malu es m ofbAFnee immaalr less 1 (Control) 2 Distilled 1-6052 water 2o 33 33 43 TT--66005522 1,200000 33 33 a Administered ata dose volume of 2.0 mL/kg. JustifforiSpceca iestSei lecotinon Hchiositcoericbaelclayu,sethoef tNheew ZlaeragleandamoWuhnitteofalbbaicnkogrroaubnbditihnafsormbaeteniontheonantihmiasl spoefcies. PROCEDURES PrepaofrEa xpotsuireo Arn ea OoTnhbetatcihlneipdupaneybdlebmeaifrseoahreemdatdesesktinu)pm,ataeptrhpieraolxfilmaaanptkpselliycoaft2i0eo%anc,hofrthatebhebibttaoctwkaalsandcb,oldiypipfesdunreffcareceseesaoarrfyeah(.atior. aTinhriermiatilansttiaowcnet,resokrcinlidpeopffeedctthsoenttheDaasytts wso8iutleadsndwpa2r9sectlioundseapiedcthteeindusvfeiosruoafilniatzneiyrnfgeorfithnteghe laepasnpiilominacsla.st.ion The sites. DoAsdeministration AmaTterainailm.alsThreecdeaiyvedofatrseiantgmleentadwmaisnisdtersaitginoantedofasthDeayres1.pective test or control Gonroeuapch 1. aniAnnali'nsdivbiodduyalweidgohste o(n2.0thenid/kagy)owfastrecaatlmceunlta.tedThaendcomnetarsoulremdatbearsieadl a(pdpirsotxiilmlaetdelwayte0r.)04wmasL/acpmp.lied evenly to the testsite at a rate of 06546 ~-- Page 11 of 41 HiT 6329-135 Groups 2,3, a(p1p-l6i0e52d) atwasa mainxded4. witFhorditshteilGlreodupwa2tearnitmoalsa dose volume of 0.01 mL/kg. The c(o2ncmegn/ktgr)a,tiotnheotfes2t00mamtgemrtial`and mixture was Stored at coms u1t,ne0dm0ip0leurtmaget/dukrget,ounrttheieslpetceatsditmviensliiyst)etseruesodif.ngtheTthheeGroatuvepessrtagm3eataenbrduilak4ldaenwnaissmiatlaysdmoi(fn20i00s.t9oe8rrcgd/nl to mdraeetsteperericnatilinveemlityxh)te.urdeoAsnweasvioncldauilmvcieduulfaaoltreddeoascfehorodfeoasctehhelaenvreielnsaple(c0tb.ia2vs0eedatndeonst1.itm0s2atebmroiidagy).weoirghtteston tdohseedaTyeveolfstwraesatm4e,nt2.5, aTnhde a10r0eacno'f, erxepsopseucrteivfeolry.theTh2e, application ranged from 0.006 to 0.024 ml/cnt. a2p0p0,roxainmdat1e,0r0a0temgo/fkg . SEeacchureadrewaitohf Elastoplast tapaappppeelrictaottaippoernovairwdaoesundcanovaleolrcecdleudsgwieisvtehadnadre1so0sv-iecnrngw.rxap1pC0eo-dlclaawrigstahuwzeSeraerbaanunsdHeadrgaetpo and restrain the animals during the 24-hour exposure period. Awrapespsrtorrxaeiimnomivanetgdelycwiotl2hl4arthsaopurawsnadtaebfratnedarandgteesdsitwsepororesacborlneetmroovpleadpemraatnedtroiwaaenllys.raepspildiucaaltiotne,st tmhaeterial ReasonforRoute ofAdministration The dermal route is a potential route of exposure in humans. ! Observations of Animals C4olbihsnoeirucvraasltiaofontbsesreranvtdaetsittowniocsre wcoea nrtderaoyclomnomdaruttceatrelidiatlyprceahddeomcsikensisatn(rdmaotraintoinna.gpparnAoddxdiimatfaittoeenrlanyloo1n,c)li2nw.ie5c,rae]and conducted daily thereafter for 28 days. mBaotdeyriwaelighatdsmiwneirsetradteitoenrmi(nDeady f1)o,r ranadndoatmiziant-iloinfeputrepronsiensatioonnDa(ybay-8,29)b.efore test ) arTedhamediininingsi)tt.riaaltiSoudnebrsmeaaqcluceonritdrirnrigetaadttioinogntsheroeDfardadiienzrgem'aylatseicmrharndiietqautbeieofn(orreweceortreedsemtdadoaers actophnpetrroDoxaliymam1tateelryia3l0 minutes after bandage removal (Day 2) and on Days 4_and 8. 0654? at Page 12 of 41 Sample Collections HHI 6329-135 bBallllooooddanwisamasamlpslceosnbdeuf(coartpeepdreoxaxppiepmrraiotnxeeilnmytaat4lelmyli)n2i4wt-eihraoetuirocsnolpl(oeDscattydeod1s).efro(mDSauybasm2e)aq,rugeinantnadlcoonlealDreacytvsieoinn4, ooffs, 2s10a8,mnpllaensodfw22eb.rleoodcIennwatasrdidfioutbgitoeandi,neadnadtfrtsohemeparttahietneedpoosiftnetnoreicorsroeprsuvymenaaonndcaDvcaaeyllo2uf9,la`reaapcfphrr.aoacxntiiimmoaanlts.e.ly"A11 T-h2e0seC 1sa0mpClesuntwielreshtihepnpedsttooredtheinaSponfsroere.zer set to maintain a temperature of Pathology wAaintdthtneerscmoridonipaustniieopdnentoinofbartrahbneidtoeamxlp,oerrdibemlree.ndtavlTihaephtsahiesteepso(sDtoafeyrit2eo9s)r,t veaannndaimcacolanvstar,woelreexmsaaatnnegerusiitanh]aetteidz.ed Aaalnplypliaacbnanitomiraomlnasliwwteeirreees wsawusebhrjeeedctreewdciotrhtdoeTdau.nkewa~abTrbhmre ewvthiaopalteewdat1iegvrreorsb,sefnboierlceer,otphseayn neeacpxrparomopixsniyantaiptorenoceadnudre. tkIii-sdcsnmueeyxs s1af-mrcpomlmessethcewteirofenirswotefigmthahelededea(rnvmdoallufmeemaaplopenlliycinadteietoacenhrmsiginrteeodupfrfwooremr'aeblillceo)lalneaicnmtdaeldsi.,mmaenTddhieatbeoltyh p"2l0acCed41o0nC.d"ry Aifcet,erthneencropplsayc,edthien aanifmraeleszewrerseetditsocamradiendt.ain a temperature of ShipmentofBlood.Bile.andTissues Afffrrtaoczeterinoenx(spo)ne,ridnrleyinvteiarclse,) tteboirlmeit,hneatdieSorpnmoa,nlsotrahepp(lJbialcmoaeostdioDs.namJpsolihtenesss,o(ns,aenrd3uMmkiaE.dnEnd.eycs&elwPl.euCrl.ea,rsBelndtg. . wd2ei-is3gp0hots9si,tioor9n3v5oolfBuumtsehhse.AvseanmTuphleee,sS.pSto.nHsNoPIraudli,osesrMNe,snopto5n5sa10ci6cb)el,pet faoalnroyngthreewistprhoentsetinhbteiiilorintcyaonrdfroersptohneding parneasleysnitsedofintthheistirsespuoerts.amples collected in this study nor are these results . Statistical Analyses No statistical analyses were required by the protocol. LocationofRawData, Records,and Final Report Trehteairnaewddaitna,therecaorrcdhsi,vesandofaHnHIoriigninaaclcorsdiagnnceed wciotphy HoMfI tShOeP.final report will be - 00548 -- Page 13 of 41 RESULTS Body Weights wIenidgihvtidugaalinsandfrmoemanDaybod1ytoweDiaghyts29.are in Table 1. HHI 6329-135 AT1 animals exhibited body Clinical Observations tIhnrdoivuigdhuoault cthleinisctauldy.signs are in Table 2. All animals appeared normal Dermal Irritation pIrnoddiuvciedudalnodedrermmaall irirrriittaattiioonn.scorNoesdearrmealiniTrarbilteati3.on wTahse animals treated with T-6052 at any of the dose levels. ocobnsterrovledmaitnertihael Patholoay oaIbnnsidmeiarvlvieddutailsinsuaeanniymwae]iogfhptatshtehoanladonigmybaillcseo.mmveonlutmsesaraerepreinseTnatbeldein5. TaTbhleere4.werIendniovidlueaslions : Page 15 contains a pathology report by the study pathologist. DISCUSSION dwTeehrremeaalceuvtaaeplpulasitycesadtteimioinnc.maalbAespopralnpidtciaoftneim/oatnloexoifaclibttihyniosanmdraatbreberilitaastliwvhedeindskiannodtmiinrrierssiuttleatrnecdyinoasafnyTa-d6se0ir5nm2galle eifrfreicttastioonn obrodytewsetigmhattergiaianl-roerlmaatecdrosicno-pliicfeficlnidniincgasl ateffneecctrso.psy.There ware no C0C549 -- Page 14 of 41 SIGNATURE HHI 6329-135 SSttuedvyenDii.recGtloarza Acute Toxicology DL ate T: S REFERENCE 1. Dorfaitzhee, SJa.feHt.y,of"ACchuetemicDaelrmsalinTFooxoidcsi,tyDr(uSgisngalnedECxopsomseurtei)c,s"- IDn:ermaAlppraisal T(o1x9i59c)i.ty, Association of Food and Drug Officials of the U.S., pp. 54-56 ) G00650 vee Page 15 of 41 HWI 6329-135 PATHOLOGY REPORT There were six rabbits (three males and three females) each from four dose levels euthanized and necropsied at the termination of the study. The test material, dose level, day of death, and gross observations recorded for each animal are in the Individual Pathology Comments that follow this report. At necropsy, there were no visible lesions in any of the animals. The liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from one male and one female in each group were cforlolzeecn,teda.nd Tsheenttitossutehe sSapmopnlseosr.werAefwteeirghneedcro(pvsoyl,umetheonlaynidmaeltsermwienreeddifsocrarbdielde.), PaH thologisw tFalmer, a (6329-135.51h) 031695 -- | a= 00651 Animal Number FF5522097729 F52073 Mean FF5522000970 F52086 Mean FF5522909926 F52084 Mean Page 16 of 41 HHI 6329-135 Individual and TMaebalne B1ody Weights (g) Male Rainzadtoino-n __Day__ Day -8 1 29 RandoFne-male Animal ization Day Number Day -8 _1_ 29 Group1 (Control) - Distilled Water (0 na/ka) 21,,099003 22,,306722 32,,054519 FF552097863 22,,115116 22,,229569 22,,688613 1,921 2,211 2,818 F52975 2,043 2,265 2,671 1,971 2,25 2,806 2,103 2,273 2,738 Group 2 - T-6052 (2 ma/ka 22,,003915 22,,325015 22,,982683 F5F25999842 12,,082825 22,,216415 22,,961841 2,034 2,332 2,816 F53410 2,220 2,471 2,875 2,088 2,29 2,869 2,02 2,202 2,823 Group 3 - T-6052 (200 ma/ka) 21,,81809 22,,300522 22,,879279 FF5522999839 21,,099973 22,,321364 22,,685714 1,050 2,257 2,993 F52977 2,049 2,323 2,687 2,010 2,204 2,873 2,06 2,291 2,737 Group 4-T-6052 (1.000ma/kg). FF5522008708 21,,198916 22,,138844 23,,066337 FF5522998975 22,,113410 22,,227449 22,,861474 F52091 2,108 2,351 3,142 F52988 2,188 2,423 3,015 Mean 2,075 2,306 2,947 _2,188 2,315 2,825 a Aunsiemailn Ntoh.e Fst5u3d4y09bwuatswaosrigrienpallalcyedseplrieocrtedtobtyretahetmernatndwoimtihzatNio.onF5p2r9o8g2ramdueforto poor health. _ COC552 m-- Page 17 of 41 IndividualTabCllieni2cal Signs HHI 6329-135 Sex Male Female Male Female Hier Observation ats Zthroudh 23 Group1 (Control) - Distilled Water (0 ma/ka) - F52979 Appeared normal v v F52972 Appeared normal v v F52973 Appeared normal v v F52976 Appeared normal v v F52983 Appeared normal v v F52975 Appeared normal v v Group 2 - T-6052 (2 mo/ka F52990 Appeared normal v v F52997 Appeared normal v v F52985 Appeared normal v v F52982 Appeared normal v v F52994 Appeared normal v v F53410 Appeared normal v v / Condition existed. 00553 -- Page 18 of 41 IndTiavbildeual2 C(lCionnitcianluedS)igns HHI 6329-135 Sex foal Male F5299 52992 F52984 Female F52989 F52993 F52977 Observation rth Group 3 - T-6052 (200 maka! Appeared normal v Appeared normal v Appeared normal v Appeared normal v Appeared normal v Appeared normal v Z Te ah 28 v v v v v v Group 4 - T-6052 (1,000 mo/ka Male F52980 Appeared normal v v F52978 Appeared normal v v ) F52991 Appeared normal v v Female F52995 Appeared normal v v F52987 Appeared normal v v F52988 Appeared normal v v 7 Condition existed. C0054 I Page 19 of 41 Table 3 Individual Dermal Irritation Scores Group 1 (Control) - Distilled Water (0 mg/kg) HWI 6329-135 Dermal Reaction Males 1 2 4 StudyDay Females & -- S LI t 2uD4ad y y & Animal No. F52979 Animal No. F52976 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 [] 0 [J 0 0 0 0 0 0 0 0 0 0 Animal No. F52972 Animal No. F52983 . Erythema 0 0 0 0 0 0 0 0 Edema 0 0 0 0 0 0 0 0 DAetsoqnuiaamat fon 00 00 00 00 00 00 00 00 CoFriissaucreionugsness 00 00 0000 00 00 00 00 Animal No. F52973 Animal No. F52975 Erythema 0 0 0 0 0 0 0 0 Edema 0 0 0 0 0 0 0 0 . Atonia 0 0 0 0 0 0 0 0 Desquamation 0 0 0 0 0 0 0 0 Coriaceousness 0 0 0 0 0 0 0 0 Fissuring 0 0 0 0 0 0 0 0 COCGSS -- Page 20 of 41 HWI 6329-135 Table 3 (Continued) Individual Dermal Irritation Scores Group 2 - T-6052 (2 mg/kg) M Say led s _F__S_etmuabvlbea_ sr Dermal Reaction IL 2 4 & LL 2 4 8 AniNom.F5a29l0 _ANno.iFm529i82l EErdyetnhaena Atonia Desquamat ion Coriaceousness Fissuring 00 00 00 00 0000 0 0 0 [] 0 0 0 0 0 0 0 0 00 00 00 00 0000 0 0 0 0 0 0 0 0 0 0 0 0 Erythema Edema DAetsoqnuiaamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52997 0 0 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 [3 0 0 0 0 Anima No.F52986 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animal No. F52994 0 0 0 0 0 0 0 0 0 o 0 0 0 0 0 0 0 0 0 0 [1 0 0 I] Animal No, F53410 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 I 0 0 0 0 0 0 0 I] COCG56 -- Page 21 of 41 HHT 6329-135 IndividuaTlablDeerm3al(CoInrtriintuaetdi)on Scores Group 3 - T-6052 (200 mg/kg) DermalReaction StMuadlyesDay L248 Aninal No. F52996 StFuednayleDsay 1248 Aninal No. F52989 EErdyetnhaema ADtesoqnuiaamation FiCosrsiuarcienogusness 00 00 00 00 o0c 0 0 0 0 0 0 o 0 o0 00 00 o 00 00 00 0 0 0 0 0 0 0 0 00 00 00 00 EEdreyntahema Atonia DCoersiqaucaemoautsinoenss Fissuring EErdyetmhaena ADetsoqnuiaamation CFoirsisaucreionugsness AnimalNo.F52992 0o 00 00 00 0 0 0 0 0 0 0 0 00 00 00 00 Animal No. F52984 00 00 00 00 00 00 00 00 0eo 00 00 00 AniNom .F5a 299l 3 0o 00 00 00 00 00 00 00 00 00 00 00 Aninal No. F52977 0o 00 00 00 0o 00 00 00 o0 00 00 00 C0655? -- Page 22 of 41 HHT 6329-135 IndividuaTlablDeerm3al(CoInrtriintuaetdi)on Scores Group 4 - T-6052 (1,000 mg/kg) Dermal Reaction SMtuadlyesDay 1. 2 4 8 FemStuday Dlayes 1 2 4&8 Animal No. F52980 Ani Nom . F5a 299l 5 EErdyetnhaena 00 00 00 00 0000 0000 Atonia Desquanation o 0 00 00 00 [0J 0 0 0 . Coriaceousness Fissuring ~~ 00 00 00 00 00 00 00 00 __Aninal No. F52078 AniNom.F5a208l7 . EEdreyntahema 00 00 00 00 0 00 00 00 ADtesoqnuiaanation 00 00 00 00 00 00 00 00 FiCsorsiuarcienogusness ~~ 00 00 00 00 00 00 00 00 EErdyetnhaema ADetsoqnuiaanat ion CFoirsisaucreionugsness Aninal No. F52991 00 00 00 00 00 00 00 00 0oo0 00 00 Aninal No. F52988 00 00 00 00 00 00 00 00 00 00 00 00 C06358 -- Animal Number F52979 F52972 F52973 F52976 F52983 F52975 F52990 F52997 F52986 F52982 F52994 F53410 F52996 F52992 F52984 F52989 F52993 F52977 Page 23 of 41 HHI 6329-135 Table 4 Individual Pathology Comments Tes Dayt Sex ~~ Died Sacrificed Necropsy Observation Group1(Control)-Distilled Water(0make) - M - 29 No visible lesions. M - HM - F - F - F - 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. 29 No visible lesions. Group 2 - T-6052 (2mg/kg. M - 29 No visible lesions. M - 29 No visible lesions. M - 29 No visible lesions. F - 29 No visible lesions. F - 29 No visible lesions. F - 29 No visible lesions. Group 3 - T-6052 (200 ma/kq' M - 29 No visible lesions. N - 29 No visible lesions. M - F - 29 No visible lesions. 2 No visible lesions. F - 29 No visible lesions. F - 29 No visible lesions. -- - Not applicable. COUG5Y -- -- foil Sex F52980 M F52978 M F52991 M F52995 F F52987 F F52988 F Page 24 of 41 Table 4 (Continued) Individual Pathology Comments HWI 6329-135 hn rificed roup 4 - T-6052 - 29 - 29 - 29 - 29 - 29 - 29 Necropsy Observation 000 mg/kq] . No visible lesions. No visible lesions. No visible lesions. No visible lesions. No visible lesions. No visible lesions. ~ Not applicable. COCSG0 Page 25 of 41 HHT 6329-135 Individual Animal TissTuaeblWeeights and Bile Volumes Animal Weight (a! Dermal Appli- Bile Sex Number Liver Kidneys _cationSite Volume (nl) Group 1 (Control) - Distilled Water (0 ra/ka) Male F52079 91.387 - 0.593 0.9 5972 83.176 16.19 0.358 115 52973 76.442 - 0.800 12 Female F52076 61.443 - 0.445 0.8 F52983 98.109 - 0.393 0.9 F52975 87.984 14.617 0.608 0.4 Group 2 - T-6052 (2 ma/ka Male F599 81.033 - 0.281 0.4 F52997 92.537 - 0.421 0.5 : F52986 83.745 16.924 0.632 11 Female F52982 84.395 - 0.568 11 F52994 83.650 15.618 0.432 1.4 F53410 91.541 - 0.38 11 Group 3 - T-6052 (200 na/kg Male F5299 78.540 - 0.673 0.46 F52992 80.845 - 0.511 1.31 F52984 89.277 15.084 [Rr 0.75 Female F52089 98.425 16.883 0.837 0.7 F52993 88.925 . 0.848 0.55 F52077 72.840 - 0.526 0.9 - - Not applicable. 00561 _ Sex Male Female Page 26 of 41 HUI 6329-135 Individual Anina) Tissue Feighisand Bile Volumes Animal Mumber F52980 52978 Fs52991 F52995 F52087 F52088 -- Veight(@)De_rm_a_l _A_pp_li_Liver Kidneys cation Site Group 4 - T-6052 (1,000 ma/ka) 83.049 15.630 0.89% 84.235 - 0.547 88.738 - 0.439 63.956 14.923 0.508 82.019 - 0.287 83.911 - 0.875 Bile Volume (ml) 1.0 1.6 0.9 0.25 1.35 1.2 - Not applicable. CO0G62 and Page 27 of 41 APPENDIX A Protocol TP3016.AB Protocol Amendment No. 1 HHI 6329-135 COCs83 -- : @ WISCONSIN ) ORRIASHIR Fh Page 28 of 41 +ConNING aroun Sponsor: 3M ToxicoloSgty. SPearuvli,ceMiMnendeiscoatla Department PROTOCOL TP3016.A8 StudyTitle: Single-Dose DermTa-l605A2bsoirnptRiabobni/tTsoxicity Study of Date: ) Decenber 30, 1994 Perforning Laboratory: ; H3a3z0l1etoKninsWmiasnconBsoiunl,evaIrndc. Madison, Wisconsin 53704 aboratory Project Identification: HHI 6329-135 J COCs6a . PEArPoRnEeSL sMoAxLE erIReAiTsVINS Shot aiigMal Bion wanFatx woE0n0 oa - Page 29 of 41 \ P3016.48 Page 2 STUDY IDENTIFICATION Single-Dose DernTa-l605A2bsoirnptRiabobni/tTsoxicity Study of HI No. 6329-135 Test Material T-6052 Sponsor : 3MDeTpoaxritcmoelnotg.y Service Medical 3P.M0.CenBtoexr,332B2ld0g. 20-26-02 St. Paul, HN 55133-3220 Sponsor's Representative 3JHohnToxL.icoBluotegnyhoSfefr,vicPehDMedical 3MDCepeanrttemre,nt.Bldg. 220-2-02 ) SPt..0.PaBuolx, 3W3N22055133-3220 (612) 733-1962 Study Director HSatzelveetnonM.WiGslcaoznasin, Inc. MPa.d0.isoBno,x W7I54553707-7545 (608) 241-7292 Study Location HBauzilleditnogn WNoi.sc3onsin, Inc. M3a8d0i2soPna,ckWeIrs A5v3e7n04ue PrEoxppoesreidneSnttuadly TSitmaerttabDlaete EDxrpaefrtinReenptoarlt DTaetremination Date JFeabnruuaarryy,3, 19199595 March 16, 1995 CO0GEs Page 30 of 41 PPa3g0e163.A8 1. SStiundglye-Dose Dermal Absorption/Toxicity Study in Rabbits 2. PTuorpaossseess irritancy otfheasytsesttemmiactearbisaolrpwthieonn aanpdpliteodxictiotytheandskirnelaotfirveabbsiktisn 3. ReTghuislasttourdyyCowimlpllibaencceonducted in accordance with the following Good Laboratory Practice Regulations/Standards/Guiindeesl: [1 Conduct as a Nonregulated Study [[X']] 2410 CCFFRR 51860(F(DEAP)A-FIFRA) [[11 C40(81C)F3R079(2Fin(aElP)A-T(S0CEAC)D) [[ 1 N5o9tiNfoihcsaatnioNno.No3.850313(J(aJpaapnaenseeseMAFHFO)HM) AW1e1lfaprreocAedcutreRseguilnattihoinss.protIoncotlhearoepiniinoncoompflitahneceSpwointshorthaendAnsitmuadly wdoirrke.ctor, the study does not unnecessarily duplicate any previous ! 4. QTuhaeliptryotAoscsoul,rasntcuedy conduct, and the final report will be audited by t(hHeI)QuaSltiatnydarAdssOuprearnacteinUgnitProicnedaucrceosrda(nScOePs)witahndHapzolliectioens.Wisconsin 5. TestMaterial A. TI-d6e0n5t2ification B. Ph(yTosibcealdoDceusmcernitpetdioinn the raw data) C. TPhe u Spaor nnsdoSi rtaasbt siulmiy etsyresponsibility for purity and stability determinations (including under test conditions). D. SRtooormagteemperature C0056 -- Page 31 of 41 TP3016.A8 E. Reserve Samles Page 4 Reserve sample(s) of each batch/lot of test and control materials will be taken for this study. The test and control material reserve samples will be stored at HWI in a freezer set to maintain a temperature of -20C $10C f10orye1a0rsyeafrosr dpiesrpoHsWIitiSoOnP. inThaeccoSrpdoannscoer wwiiltlh btehecoanptparcotperdiataefter regulatory Good Laboratory Practices. F. ARneytenutnuisoend test material will be returned to the Sponsor after completion of the in-1ife phase of the study. 6. SafetyPrecautions As required by HWI SOPs and policies 6. ControlMaterial A. Identification Distilled water -) B. PhCylesairc,alcoDleosrclreispstiToinquid C. puarndSitabtiliyty Tchoensipduerrietdy atondbestaadbeiqluiattyeofforthitshemapnurupfoascetsureodf mtahtiesrisatludyi.s ! D. StorageConditions Room temperature E. Reserve Samples See Section 5. E. Reserve Samples F. Retention Any remaining control material may be used for other testing and will not be discarded after issuance of the final report. 6. SafetyPrecautions . As required by HWI SOPs and policies 7. Experimenta)Desian A. Animals = (1) Soecies Rabbit J (2) Srtar:ai(nN/ZWS)oSuPrFc/eHRP, Inc. J C0567 -- Page 32 of 41 *y 3) AIgenaittiation (4) M2.0et0 ai3t.I0ngikgtihatiton PTaPg3e016.AB (5) N1u2mbmearleasndanSdex12 females . (6) IIdnednitviidfuiaclatniuomnbered ear tag (7) Husbandry (2) HIonudsiivindgually, in screen-bottom stainless steel cages (heavy gauge) (b) FAomoedasured amount of Laboratory Rabbit Diet HF #5326 t(hPeHImaFeneudfsa,ctuIrnecr.).forTnhuetrfiotoidoniasl rcooumtpionneelnytsanaanldyzed by environnental contaminants. ! (c) M4dateTribitum from an water are analyzed abyutWoMmIatifcorstyosttaelm. disSsaomlpvleeds osoflitdhse, hfaorrdnseeslse,ctaenddelsepneecnitfsi,edhmeiacvryobmieotallosg,ical content and organophosphates, and' chlorinated hydrocarbons. (d) CTohnetreamianreanntos known contaminants in the food or water that would interfere with this study. (e) EEnnvviirroonnmmeennttal controls for the animal set to maintain a temperature of 19C rtooom23wCi,ll abe rTeilgahtti/v1e2-hhouumriddiatrykocfyc5l0e%.420%, and a 12-hour (f) AAtcclTiemasattio7ndays - (8) SBealseedctioonnhoefaltThestandAnibmoadylsWeight according to HI SOPs. An naodeqauniamtaelnuinnboerbvoifouesxltyrapooarnimhaelaslthwilils pbleacpeudrchonasetdests.o thTahte ) santirmaatlisfiewdillbodbye placed weight rianntodomsitzuadtyiognroupprsogrusainngwitahin nine - days of study initiation. _ COCsE8 -- Page 33 of 41 \ 1P3016.A8 Page 6 (9) JustifforiSpcecfaestSeilecotinon Historically, the New Zealand White albino rabbit has been the animal of choice because of the large amount of background information on this species. 8. DAodsneinistration (1) TestGrows Group TestMaterial _D_o(senaL/evkeal) HalNesuogf AbFneimemaallress 1 (Control) Distilled water [9 3 3 2 T-6052 uid 3 3 3 1-6052 200 3 3 4 T-6052 1000 3 3 * To be administered at a dose volume of 2.0 mL/kg ** To be administered at a dose volume of .01 mL/kg (2) POn rtheedpayaboefrfEorxaepottseusirteAmoartneeraial application, the back and, if necessary (to obtain unblemished skin), the flanks of ) each rabbit will be clipped free of hair. The shaved area will constitute approximately 20% of the total body siunrsfpaeccetedarefao.r iTnhteertfreeriantgmenltesisointse,s (irirnittaacttiosnk,in)orwidlelfecbtes that would preclude the use of any of the animals. The animals will be clipped as needed throughout the study. (3) DoAsdneinistration A101 animals will receive a single administration of the respective test or control material. The day of treatment will be designated as Day 1. The dose for each animal in Group 2 will be diluted with distilled water and applied at a dose volume of .01 mL/kg. The respective dose for each animal in Groups 3 and 4 will be applied undiluted. Al] doses in Groups 1-4 will be based on the animal's body weight just before administration and will be spread onto the area of exposure in a thin and uniform a layer. The area of application (Groups 1-4) will be covered with a : 10-cn x 10-cn gauze bandage secured with paper tape around, all edges and overwrapped with Saran Wrap' and Elastoplast' tape to provide an occlusive dressing. The rabbits will be collared during the 24-hour application period. (4) Reason for Route of Administration The dermal route is a potential route of exposure in humans. - J = CO0GEY -- Page 34 of 41 !) TPPa3ge0167.A8 (5) AR apppprle oixciam tmoiafotneTo leytshtv e24Mbaahta noeduraril gsaeslaftaenrd ctoelsltarosr wciolnltroble mraetmeorvteadl and atnheaprpesriodpurailatetesstolmvaentte,riailf wnielclessbaeryr.emoved using water or C. Observation of Animals (1) ClFaodirmniicncliaisnltirOcabatslieornsviagatnnisdonbfseofrorcelintiecsatl osrigcnosntraonld mmaotretrailailty at asaidpgnpnirsno,ixsitnarnadatteiltowynic1e,(Dd2a.ay5i,l1y) ana(dnad.m4.dahiaonludrys pt.hamef.rt)eearfftotreersmtofrmotaratelcrilitiany)icfaolr daitrelcetaesdt 2b8y dtahyes.studOybsedirrveacttioorn.s may be extended When (2) RdeBeearfdnoiarnlegoLierfsrtiDteaortrmiaoclnonItrrreroaildtianmtgaitoewnrililalbeadmmaidniestarnadtiroencortdheediansititahle Daifartyreirt1artbeaianoddniangrgeeadr(ieAnmtogtvsaaclwhiule(lnDtaybe12)).madaAendddaioptnpiroSontxauildmyatdDeeralmyyasl304 mainndut8.es ) fIonrdievaidcuhalanidmearlm.al irritation records will be maintained (3) BFaoopdrpylirWcaeanitdigoomhnitzsa(tDiaoyn,1),beofnoreDayte2s9t,oarndconattroulnscmhateedruilaeld death {when survival exceeds 1 day) (4) sampleCollections -. (a) BEerfeoqrueenciynitiation post-dose (Day 2), (DDaayys 1)4,, 8a,ppr15o,xim22a,telaynd 24at hours experinental termination (Day 29) (b) ManusofbAneimarls ) CO0G70 Page 35 of 41 TPaPg3e0186.48 (c) BfMlrooone dtosht eafmmCpaloh relgslieo n(caatlpid poernaorxivmeaitneloyf4eimtL)herwilelarboencoDlalysect1,ed 2(,ac4t,ual8, vo15l,umeandto22b.e doAcpupmreonxtiemdateilnyth2e0 nrLawodfatbal)oowdill abenimoabltaisnaecdriffircoemdthine paomsotreirbiuonrdvcenoandictaivoanofore-ach ssaamcprliefsicweidllatbethestotriende of at rnoeocmroptsemype(rDaatyur2e9).andThtehen cfernatcrtiifoungseds,toraendd itnheasferpeaerzaetre sseetruntoamnadinctealilnular f-r2a0ctCio$n1s0Cw.illThbee sseenptaraftreodzenseornumdrayndicceelltuolatrhe Sponsor after experinental termination. Samples will be shipped to: 3JMameEsED. &JPo.hCn.son 9B3l5dg.Bus2h-3Av-e0n9ue St. Paul, MX. 55106 ) JtaenleesphoD.neJoathns(o6n12)or77a8l-t5e2r9n4ateprwiiolrltobetnhoetisfhiiepdmenbty of the samples. 0. Pathology (1) UAnnsychaneidmualleddySiancgridfiucreisngatnhdeDesattuhdsy or sacrificed in a mgroorsisbunndecrcoopnsdyitieoxnamwiinlaltiobne asnudbjaelcltedabntoormaanliatbibersevwiialtledbe - raenceosrtdheedt.izedAnwiimtahlssoidniuanmopreinbtuobnadrbciotnadliti(ovniawiilnljebcetion in the marginal ear vein), bled via the vena cava, and eD.xsaPnagtuhionlaotgye,d. (3)TisSsaumepsl,e Caoslldeecstciroinb,edwiilnl sbeectcioonllected. After necropsy, the aninals will be discarded. (2) SActhetdeurlneidnSataicornifoifcethe experimental phase (Day 29), pseunrtvoibvairnbgitaanlina(lvsiawiilnljecbteioannesinthettheizpeadrgwiintahl seoadriumvein), balbebdrevviiaatetdhegrvoesnsa cnaevcar,opseyxsaenxagmuiinnaattieodn;. andThesuabnjiemcatlesd wtiollan bbee nreeccorrodpesdi.ed in random order and all abnormalities will J) Ccocs7L -- Page 36 of 41 ) TPaPg3e0169.A8 (3) sTahmeplseiteCsololfectteisotn and control material application will be wparsohceeddurwei.th TlhuekewahrolaetaTpvewra,terbilper,iorantoaptphreoxinmeactreops1y-cm x aIn-icmaalsse,ctiaonnd boofththekiddneremyasl fraopmpltihceatifoinrstsimtealefraonmdalflemale npleaccoepdsieidn ainferaecehzergrosuept wtiollmaibnetacionlleactteedmpaenrdatuirmemeodfiat-e2l0yC 10C. After necropsy, the animals will be discarded. Tkhiednetyiss)suewisll(lbievers,entbilfer,ozdeenrmoanl drayppliicecattoionthesitSep,onsor asfhtieprpedexpteoritnheentpaelrsotnernliinsatteidoni.n STehcetiosnam7p.lCe.s(4w)i.l(lc)b.e The oSfpontsheorsaimsplreess.ponsible for the retention and disposition E. StWoatsitsattiicstailcAanlalaynsaleysses are required. 8. RAepfoinratl report including those items listed below will be submitted. ) DDeessccrriippttiioonn ooff tthhee tteesstt sanydstecmontrol materials DPartoecseduorfesexperimental initiation and termination DTaebsuclraitpitoinonofofmoarntyaltiotxyicdaetffaecbtys/sdeexrnaanld diorsreitaltevieoln GTraobsuslatpiaotnholoofgymeafnindbiondygs/wgeriogshstspbaythoslexogyandrepdoorste level 9. LoOcriagtiinoalnodfaRtaa,woDratcao,pRieescotrhdesr,eoafn,dFwiilnlalbReeapvoaritlable at HHI to afcacceipltiatnactee oafuditthiengfintahle rsetpuodryt.duriWnhgenittsheprfiongarlessreaponrdtbeisforceompleted, r31e1taionreidginianlthpeapearrchdiavtae,s oifncHlAuIdinagccotrhdoisneg ittoemH1I1sStOePd. below will be PDroosteocporlepaarnadtipornotorceoclordasmendments : In-BToidfyeweriegchotrsds ODbosseervaadtmiionnisstration ~~ SAnaamtpolmeiccaolllepcattihoonlogryecorredcsords SSthuidpypincgorrreescoprodnsdence ) Final report (original signed copy) = COGST2 Page 37 of 41 ) PTaPg3e01610.A8 Tnohte bfeollaorwcihnigvedsuwpiptohrtitnhge rsteucdoyrdsdatwai.ll be retained at HMI but will AWnaitmearl anraelcyesiipst/raecccolridmsation records ARneifnrailgerraotoomrteanmdpefrraetuerzeeratnedmpheurmaitduirtey rreeccoorrddss . Instrument calibration and maintenance records ) , - COG873 - Page 38 of 41 ) TPaPg3e01611.A8 PROTOCOL APPROVAL Fm 2. QrZentsly John L. Butenhoff, PhD Sponsor's Representative 3M Toxicology Service Medical Department som Date Steven H. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. ) ld. presentative Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-135.protdsk2) 230 ay Date : `also [94 Date ) - COCG 7a Page 39 of 41 TP3016.AB ) Page 12 Attachment 1 Scoring Scale for Acute Dermal Reactions Erythema 0 - None 1 - Slight 2 - Moderate 3 - Severe Edema 01 - - None Slight (barely perceptible to well defined by definite raising) 2 - Moderate (raised approximately 1 mm) 3 - Severe (raised more than 1 mm) Atonta ) 01 -- NSolnieght (slight impairment of elasticity) 32 -- HMoadrekreadte(no(selloawstriectiutryn) to normal) ! Desquanation 0 - None 21 -- MSoldiegrhatte(sl(isgchatlesscaalnidngf)lakes) 3 - Marked (pronounced flaking with denuded areas) Cortaceousness 0 - None 1 - Slight (decrease in pliability) 2 - Moderate (leathery texture) . 3 - Marked (tough and brittle) Eissuring - 0 - None - 21 -- MSoldiegrhatte(de(fcirnaictkes icnradcekrsmiisn) epidermis) 3 - Marked (cracks with bleeding) ) C0575 _ Page 40 of 41 yo @ PRWOSSITBSLOFCSFIROCEINNBoSSx I7B3N4i3 +common PROTOCOL TP3016.A8 Single-Dose DoefrnTa-l605A2bsoirnptRiaobnbi/tTsoxicity Study HHI 6329-135 . Sponsor 3MMeTdoixciacloloDgeyparSterrevnitce 3P.M0.CenBtoexr,332Bl2d0g. 20-26-02 St. Paul, MN 55133-3220 Contractor H33a0z1letKoinnsmWainscoBnosuilne,varIdnc. Madison, WI 53704 Sponsor'sRepresentative John L. Butenhoff, PhO StudyDirector Steven M. 6laza Anendnent No. 1 : This amendment modifies the following portions of the protocol: ) Effective January 24, 1995 Aotf bthielerceoqluleesctteodf wtihlelSpboensdoorc,umetnhteedweiignhttsheofrawtisdastuae.s coTlhleseectewdeigahntdstahnedvolume ! svoelcutnieosnswiolfl thbee pirnoctloucdoeld wtiothinctlheudesatmhpelseesahdidpimteinotn.s.Modify the following I. PHaogdeiFy9, Th7.e sEexcpoenrdimesnetnatlencDeesiiann;theD. FPiartshtolaongdy:seco(3)ndSapmaprlaegraCpohlslecotfiotnhis section with the following underlined additions: dTehremawlholaepplTiivceart,ionbilsei,te anfroanpparlolximaantineal1s-,cmanxd 1b-octmh skeicdtnieoyns offrotmhe ctohlelefcitresdt, mwaleeighaendd f(evmoalluemenoenclryopdseiteedrmiinneedachfogrrboiulep),willandbe . -i2m0meCdi$a1t0elCy.placed in a freezer set to maintain a temperature of Tshheippseadmpltoesatc hnedpto ehresr iorn r lise tedsin p Seco twieoin ngh7td .sCo<(i r4v).on (lcu)mg .es will be 2. Page 9. 8 Report. Add the following to this section: Individual animal tissue weights and bile volumes . Phone Gon 241 aes COC576 Fax sow rar izes -- --_-- EArt rit BLIERT IRI NEHAN YE AIS ayTasV2 Page 41 of 41 ) Amendment No. 1 HPWaIge 632 29-135 PROTOCOL AMENDMENT APPROVAL Fl 2 Bulent) John L. Butenhoff, PhD Sponsors Representative 3M Toxicology Service Medical Department 2 HSH Date Wee NN Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. Ne = ) Representative) Quality Assurdnce Unit Hazleton Wisconsin, Inc. (6329-Am1l .3ds5k2.) Date 2-6 2.2.95 Date wns. ; CoesT? -- 9.1.2 Analytical protocol AMDT-020795.1 CO0G78 3M Environmental Laboratory Protocol - Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T-6052 in Rabbits In-Vivo Study Reference Number: HWI#6329-135 Study Number: AMDT-020795.1 Test Substance: FC-120 (T-6052) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facility: 3M Environmental Technology and Services 935 Bush Avenue St. Paul, MN 55106 Proposed Initiation Date: July 25, 1995 Proposed Completion Date: August 25,1995 Method Numbers and Revisions: AMDT-M-1-0, Thermal Extraction of Fluoride by Meansof a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver AMDT-M-20, Fluoride Measurement by Meansofan Orion EA940 Expandable Ton Analyzer AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver AMDT-M-5-0, Analysis of Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry AMDT-M-8-0, Analysis of Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode AMDT-M-14-0, Thermal Extraction of Fluoride by Means ofaModified Dohrmann DX2000 Organic Halide Analyzer-Serum Author: James D. Johnson fps Approved By: mes D. son tudy Digector Date FL o LL fini _ John Butenhoff, PhD Sponsor Representative: Date _ C0657Y -- 1.0 PURPOSE dTheirsmasltluydyabissodrebsedi.gneTdhetoapnraloyvtiidcealinafsopremcattoifonthaiss to whether study is to FC-120 (T-6052) is determine fluorine- ocobnttaaiinneidnagftceormdpeorumnadlsabisnotrhpetiloinvewriatnhdtshaetruobmtoafinreadbbaifttse.rBiyntcroavmepnaoruissoinnjeocftitohne, data assessment of the extent of dermal absorption can be performed. 20TESTMATERIALS ~~~ 2.1 Test, Control, and Reference Substances and Matrices 2.1.1 Analytical equivalent. Reference Substance: FC-95, lot 161 or 171. They are 2.1.2 Analytical serum Reference Matrix: Bovine liver, bovine serum and rabbit 2.1.3 2.1.4 Analytical Analytical Control Control Substance: None Matrix: Bovine liver, bovine serum and rabbit serum l2i.v2erS),ouSricgemaofCMhaetmeirciaallsC:o3mpMaInCyP/(2P.C1.P2,Di2v.1i.s4iobnov(2i.n1e.1s)e,rgurmo)c,eAryMsDtoTre 1(12.013.924,.21.1.4 (Hwi#6329-123) control group animals (2.1.2, 2.1.4 rabbit serum) 2a.n3dN6ucmonbterroloafniTmeaslts.anTdisCsounetsraonldSfalmupildsesi:ncTliusdseuleisvear,ndkifdlnueiyds, fsreorumm,18cetlelstulaanrimals dfirsacctrieotni,ondeorfmtahleaSptpulidcyatDiiorencstiotre.and bile. Analysis of these tissues will be at the t2h.e4 HIdWenItiafniicmaatlioindeonftiTfeisctataionndnCuomnbterrolwhSiacmhplceonss:isTthseofsaamlpeltetesraarnedifdievnetidfiigeidtusing number, plus the tissue identity and day identity (serum). 2.5 Purity and Strengthof Reference Substance: To be determined by Sponsor. 2.6 Stability of Reference Substance: To be determined by Sponsor. 22.07 %Sto1r0agCe(C2.o1n.d2,it2i.o1.n4s).foTreTsetsatnMdaCtoenrtiraolls:saRmopolems temperature (2.1.1), will be received according to AMDT-S-10-0. ~- G2.L8PDiRsepgousliattiioonnofforSptehecitmiemnesp:erBiioodlorgeiqcuailretdisfsouresstaunddiefsluliodnsgweirllthbaen r2e8tadianyesd.per _ COGSB0 ss 2.9 Safety attire. Use Pcraeuctaiuotniwohnse:n Rheafnedrlitnogakpnpirvoepsrifaotre cMutStDiSng.theWesaamrplaepspr.opriate laboratory Tanhaelytsiisssufeosrffrluoomrianneimcaolmspoduonsdesd.asAdtestchreidbiesdcr(eHtWiIo#n6o3f2t9h-e13S5t)ud,yaDrieraevcatiolra,balesefroires of a(nSaeleytMiectahlotdess-ts-nceaxntbSeecpteirofno)rmisedt.he Tahpperospcrrieaetneinagnafloyrsifslutooripdreesiennltivdeerfivniiaticvoemdbautsatfioorn pfrleuosreinnteiinnltihveer(liifvearn. yToatc2o8nfdiaryms)thaeniddesnetriutym oatf vflauroiroiunsei-nctoenrtvaailsn,inegleccotmrposopurnadysmass sopfetchtertoimsestureys may and fbleuisdeslaemctpeldesaswiolnleobfetahnaelyaznaeldy.tiWcahletnecshunfifqiuceiesntemdpaltoayheads. beNeont all canoalllyescitsedwitlolmceeeaste.the objectives of the study in the opinion of the Study Director, 4.0EXPERIMENTAL-Methods 4.1 Liver and Serum screening methods: (attached) 4D.o1h.1rmAaMnDnT-DMX:2-010-00,OrTghaenrimcaHlaElxitdreacAtniaolnyzoefrF-lLuiovreirde by Means ofaModified 4E.x1p.a2ndAaMbDlTe-TMo-n2A-n0al,yFzleuroride Measurement by Means of an Orion EA940 4.1.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver 4U.s1i.n4gAEMlDecTt-roMs-p5ra-y0M,aAsnsalSypseicstroofmReatbrbyit Liver Extract for Fluorochemicals 4A.n1a.l5yzAeMrDwTi-tMh-I8o-n0S,elAecntailvyesiEsloecftrFoldueoride Using the Skalar Segmented Flow 4M.o1d.i6fiAeMdDTDo-hMr-m1a4n-n0,DXT2h0e0rm0aOlrEgxatnriacctHiaolnidoef Fluoride by Means Analyzer-Serum ofa 00681 a f5l.u1oDriadteapeRreptoirsstuiengor: flDuaitd,aowrilalsbFeC-re9p5or(teeldecatsroascproanycemnatsrastisopnec(twreoimgehttr/yw)eipgehrtu)niotfof rtiesgsrueessoironflauinda.lysSitsaotifstsiecsruusmedc,onactenthteradtiisocnrsetwiiotnhotfitmheeaSntdudayveDriargeecstoarn,dmsatyanidnacrldude sdteavtiiasttiicoanlsteosftscosnuccehntarsaStitoundsenfotr'tsditfefsetremnatydobseeagprpoluipesd.tIofdenteecersmsianrey,stsatiimsptilceal difference. 6.1 Raw approved Data final raepnodrtR,eacnodrdesl:ectRroanwicddaatta,a waiplplrboevemdaiprnottaoicnoeld, ianpptrhoepAriMatDe Tspaerccihmievness., 2~ 70REFERENCES ~~ 7.1 AMDT-S$-10-0, Sample Tracking System 8A 0 TTAC~ H ~M ~ ENTS D8.o1hArMmaDnTn-MD-X12-000,0 Thermal Organic EHxatlriadcetAionnaloyfzeFrlu-oLriivdeer by Means of a Modified E8.x2paAnMdDaTbl-eM-Io2n-A0n,alFylzuoerride Measurement by Meansofan Orion EA940 8.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver E8l.e4cAtrMoDsTpr-aMy-M5a-s0s,SpAencaltyrsoimsetorfyRabbit Liver Extract for Fluorochemicals Using w8.i5thAIMoDnTS-eMl-ec8t-iv0e,ElAencatlryosdiesof Fluoride Using the Skalar Segmented Flow Analyzer D8.o6hArMmaDnTn-MD-X1240-000,OTrhgearnmicalHaElxitdreacAtnioanlyozfeFrl-uSoerriudme by=Meansof a Modified 00582 ae 3M Environmental Laboratory Method Thermal Extraction ofOFrlguaonriicdeHablyidMeeaAnnasloyfzaerM-oLdiivfeired Dohrmann DX2000 Method Identification Number: AMDT-M-1 Revision Number: 0 Adoption Date: /o- #-1 Revision Date: None _-- Author: Rich Youngblom `Approved by: ~ fn-- ifoup Leads /fo/3/3< Date LiphVosrmix Quality Assurance f2o-g4-7 Date Software: MS Word 5.1a Affected Documents: AMDT-M-2 Fluoride Measurement by Meansof an Orion EA940 Expandable fon Analyzer = AMDT-EP-3 Organic Routine Maintenance Halide Analyzer ofa Modified Dohrmann DX2000 0C5583 1 . = L0SCOPE.APPLICABLECOMPOUNDS.ANDMATRICES fw1li.u1tohSraicndoepfefo:rnoTsmehlivesacrtmiievoeutshelomeadcttrirsiocfdeoesr..thTehoepefrlautoiroidneofisatyDpiocharllmyacnonllDecXt2e0d0in0TwIhSeAnBitsiosluutsieodn tfoorexatnraalcytsis 11.32 MAaptprliicceasb:leBiColoomgpicoaulntdisss:ueFs,lupoarrtoicchuelmairlcyallisveorr, other fluorinated compounds. '2.D1X2F0l0uo0r,idfel,uofrloucorhienmei,caexltsr.action, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, 33..12 GGllaasssswwaarree aisnfdraegxihlea,usbtrogkaesnesgclaasnsbmeaeyxtcraeumseelyinhjoutr.ies 333...543 PPSorotelesvnsetunirtailzbebaidsoeghdaaszseaasrm,dpsplredosupeemratocyothfmleparbsieh,oslsmoegadiycganalesmeahdtarntiodcleaisln.lgoUwpsrtaehcteaipmcpertsoodprrerqiyuaidtreoedpw.enrsboenfaolrpersottaercttiinvgerun. equipment. 4.0 SUPPLIES AND MATERIALS 44..12 CCoommpprreesssseedd HOexlyiguemn,,HHiygdhroPcurairtbyoGnrfardeee,, rreegguullaatteedd ttoo3405PPSSIL, 44..34 QQuuaarrttzz ggllaassss csoammbpulsetbiooanttwuibteh, TReelfilaoncneTMGtluabsisng,G-D9o4h0r5m-a0n1n2o8r90e-qu0i9v7aloernte,quivalent, 4.5 4.6 SOarimopnle94c0ol9l9e9ctTiootnavliaIlosn,iHcDSPtrEe.ngth Adjustment Buffer (TISAB 11) or equivalent, 4.7 Milli-QTM water 44..98 PAocltyisvtaytreednCehapirpceotatle,s.E. Merck 2005 or equivalent, 44..1110 MHiasmcielltloanneSoyursinlgaeboorrateoqruyivgallaesnts.ware 5.1 52 RIoBsMemcoomupnattiDbolher3m8a6nnorD4X8260c0o0mpOurtegra.nic Halide Analyzer, modified for fluoride extraction. 5:3 5.4 EDxXc2e0l0S0prseoafdtswhaereet,,vveerrssiioonn 15..000,ormgordeiaftieerd for fluoride extraction. 60INTERFERENCES ~~. 6m.a1ySvaamrpylferosimzemaitsrliixmittoemdattoriaxp.proximately 150 mg, depending on sample moisture content, This C00384 2 7.0 SAMPLE HANDLING s7a.1mpSlaem.plUesseafroerncoetpstoorbeprhoabnedlteodtrwaintshfebratriesshuaensd.s. Fluoride may leach from the skin to the 7.2 Samples ofliverare cut from frozen liverand placed ina tared and labeled weigh boat. Use a clean scalpel and cutting board. The cutting board and scalpel should be cleaned with water, methanol, or methanol-water solution after each liver is cut. 8.0 CALIBRATION AND STANDARD) ZATION 8.1 Preparation of Calibration Standards R8.e1f.e1rTthoeprsottaoncdoalrdfsorrtehqautiprreodjefcotr.each project will need to be appropriate for that individual project. 8.1.2 Typically 50-500 ppm FC-95 in methanol standards are used. 81.510.3mgFo)rarnadbbwietilgihveritsitnudaileasb,eulseedbaenedf liver tared as the weigh matrix. boat. Cuta pieceoffrozenbeef liver (100 - 8.2 Calibration - Overview c`uThrevenoisrmraeqluciarleidbrtahteipornoicsetdhuerfeluliosrtieddebceulrovwe i(sAuMsDedT.-M-2). However,if an optional spiked liver 8.2.1 A calibration curve for the DX2000 is generated by spiking samples with known standards 8a.n2d.2coTympbiucsatlliyn,gtthhreeemruespilnicgattheseofseamaechmestthaonddasradnadndmaftirviexcotnycpeenatsratthei.onssaomfpslteasndtoarbdestewsitleld.be. spiked. aR8n.e2dc.o3tvheeSrt1ea2dnvdFaal(ruuedg. c)uornvethweilyl-abxeisp.lotGteend earsaMtaaessreSgrpeiskseidonFc(uurgv)e oanndthcealxc-ualxaitse atnhde Standard Mass equationforthe line 8.2.4 Mass Spiked F (ug) = (Amount spiked in mL) x ( Conc. ofstandard in ppm) x (0.6004)* *FC-95 is 60.04%Ftherefore 0.6004 is the factor usedto convert FC-95 toF 8.2.5 Standard Mass Recovered F (ug) = (TISAB volume in mL) x (Orion reading in ppm) 8.3 Calibration - Procedure 83.1 Start Up 8.3.1.1 Run 2 or more `be usedifthe previous sCalmepalneCsyccolnetsaiwnheednhistgahrtcionngceinntsrtartuimoennstofeafclhuodraiyd.e.More clean cyclesmay 8.3.2 Blanks 83.2.1 Prepare sample using the same methods and typeofmatrix as the test sample. 8.3.2.2 For rabbit studies, usebeef liver as the matrix. Prepareat least3 samples ofbeefliver (100- 150 mg)forblanks, 8.3.2.3 Put sample in Dohrmann boat. Combust each sample as de=scribed in section 9.0 and. analyze sample according to method AMDT-M-2 for the ion selective electrode analysis, COGG85 3-- Jb8u.ed3f.og2re.em4eprnFotocorefteradhbiebnigtopwseitrutadhtioetsrh,etcthahelerimbereaatdtieinrognr.iesaBsdtiuanmbglesfoawrmiptalhbelrseasnupnketicsltatmohpilsheilsistmohirotiiuclaldrrebeaeacd0hie.nd0g,3sop(rppurmnetvoliroilunoswt4eher hours). 8.3.2.5 For non-rabbit before proceeding with studies, the blank the calibration. readings should reach a predetermined ion concentration c8o.3m.b2u.s6tiIotnm.ay be necessary to mix approximately 50 `mg ofcharcoal with the sample to aid 88..33..33.1StWaenidgahrdoutCuartlveeast 15 ma labeled weigh boats. For rabbit tsruidxicssa,mpwleeisgh(51s0t0a-n1d5a0rdmsgwbietehf3 lrievpelricsaatmepsleesa.ch)Reincotrardedweaingdhts in 8st3ud3y2dataP.lacSetowreeitghheemdabtereifxlsivaemrplseasmopnledriyn Dicoehorrmiacneapascakmspltoe kboeaetp. them frozen until used. 83.33 Start with the lowest standard concentration. Using a Hamilton syringe, ejecat fixed quantityofthe standard on or in the matrix. For rabbit studies, us4e uLofstandard and eject iton 8or3i3n 4theAbteleefaslitve3r.replicates should `may be used at the discretionofthe baneaulyssetd. for the lowest standard concentration; more replicates 8.3.3.5 Combust the sample as described in section 9.3 and analyze according to AMDT-M-2. 8.3.3.6 Run all 15 standards.Ifonereplicate is significantly different from the other two replicates, run another sample for that standard. Indicate in data that the new replicate replaces the old replicate and 83.3.7 When all that the new replicate standards have been will run, be used to calculate calculate the 12. r2 the: regression curve. must be at least 0.95. Ifit is not at 8le3a.s3t.08.95A,nceonwsuslttanwdiatrhd changed. New standard scupuerrvveissohro.uldberun w curve may also be run haet nthtehdeicscormebtuisontoifotnhteubaenaolry ssta,mple matr ix is 8.4 Storage Conditions for Standards 84.1 Storage requirements for standards are dependent on the individual standards used. Typically, standards are stored at room temperature in plastic screw top bottles. 8.4.2 New FC-95 standards should be prepared at least once a month. 9.1 Typical Operating Conditions: 9.1.1 Combustion tube temperature = 950C. 9.1.2 Oxygen and Helium flow = 50 cc/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 Open the SYSTEM SETUP window. - 9.2.3 Put the fumace module and the cell in the READY mode. 9.2.4 Close the SYSTEM SETUP window. 00686 f= 9:25 found When in the tCheEoLvLenChHaEs CreKacmheendut.he READY temperature, run the CLEAN BOAT program 9.2.6 See AMDT-EP-3 for detailsofthe Dohrmann software, m99.3i3.x1SaapOmpprpeolnxeitmEhaxetteSrlaAycMtSiPOoLnmEgPorHfoccAheTadrCucrHoea:alndwiptlhactehethseamspalmeplteo aiindtcheomBbOusAtTi.on.ItImfathyisbeisndeocnees,sacrhayrtcooal 9s.h3o.u2ldCallossoebeSAmiMxPedLEin HwhAiTleCHes.tablishing the baseline and when generating the standard curve, 2l9a..b30e.3mleLdAosdfadm1pa:pl1peTrocIpoSrlAlieBact:teMioivnollvliuiaml-.eQoTMTfywpTaitIceaSrlAlmByix0st.ou6lrumet.iLontoor151:m1 LTIaSrAeBu:sMedi.llFio-rQrTMabwbaittesrtumdiixest,urueseto1.a0 or i9n:c3h.e4s.PlGaacseetsherevliealasseodtdhuartitnhgeptyirpoolfystihsemCuOstMBbuUbSblTeItOhrNouTgUhBthEeiTsIiSnAtBh,e TISAB at least 0.25 9.3.5 9.3.6 93.7 IWRfuhunnednithlteuhteEeEOdXOT-IXSSpOArLBoIgwDraaSsmpuirsseofdgirntiaosmhceofdlo,leurcntedmtiohnevetshatemhpRelcUeo,Nllaedmcdetniaounn.eviqaulalfrvoomlutmheeocfomMbiulsltiio-nQTMtuwbae,ter 9to.3t.h8eRTiInsSeAtBhetoenmdaokfeth1e:1 cToImSbAuBst:iMoinlltiub-eQTMw,ith remove any TISAB remaining on the tube. Milli-QTMwater and wipe with 2 KIMWIPE to `9w.i3t.h9a Ocpotetnonthteipspaemdpalpeplhiactacthoraonrd vraecmuouvmeeadnoyutr.emIatimnainygbaesnhefcersosmarthyetbooastc.rapApsahrtciaclnesboefrfetmhoeved. bbooatttotomawiidtihnathsepaCtluelaanoroCytchlee.r similar device. A drop of Milli-Q water may be added to the 93.10 9.3.11 CRluonsethteheChLatEchA.N BOAT program. 9.3.12 Sample is ready for analysis by ion selective electrode (AMDT-M-2). 9.4 Sample Calculations 9.4.1 9.4.2 SUasmeptlhee sMtaasnsdaRrdeccouvreveretdo cFal(cuugl)a=te(tTheISsaAmBpvloelvianlume.L) x(Orion readinginppm -intercept) (Slope) 0 VALIDATION 1100..11.Q1uDaaliiltyyStCaornttrUoplCheck Samples: Once the standard curve is established, each day of canoanlcyesnitsraitsisotnarstpeidkebdysaanmaplylzeisngusQedCtosagmepnleersa.teTthhee sQtCandsaarmdplcuersvea.reEtaocbhectohnecsenatmreataisonthmeulsotwebset rdeopnleiciantetriispnloitcanteecuenslseasrsy.the first two replicates are within 20%ofthe standard curve, then a third F1l0u2oriPdreecNiostieobnooaknsd2A,c3,cuarnadcSy.: PSreeceimsieotnhaondddaecvceulroapcmyenvatriaensalwyhseisnaanndalsyazmipnlgesaanmapllyessiosfidnifferent `matrices and different reference compounds. 10.3 Other Validation Parameters: NA 00687 so. 11.0 DATA ANALYSIS 1L1 Calculations 11.1.1 For the standard curve, use regression analysis in Excel, version 5.0 orgreater. 11.1.2 To calculate the fluoride contraction in the sample, see method AMDT-M-2, 11.2 Analyzing the Data a1r1e.2w.i1thi2n m2u0s%t bofeaetalcehasotth0e.r9a5nodrtghreeaotuetrl.ie"rOuitslgireerast"ermtahyanb2e0e0x%colfutdehdeifavtewroaogfetohfethtohsreeetrweoploicrates. less than 50%ofthe averageofthose two. Any such noted in the Final Report along with the reason it was outliers should be pointed consideredan outlier, out in the data and 120 ATTACHMENTS ~~~ None 13.0 REFERENCES 31439.1, rReovsiesimoonuBn,tDDeochermmbaenrn1D9X932)000 Organic Halide `Analyzer Operator's Manual (Manual 915A1n3a.2lyzAeMrDT-M-2 Fluoride Measurement by Meansofan Orion EA940 Expandable lon A1n3a.3lyAzMerDT-EP-3 Routine Maintenance ofa Modified Dohrmann DX2000 Organic Halide 14.0 REVISIONS Revision Number ReasonforChange Revision Date COGS 6 oo 3M Environmental Laboratory Method Fluoride Measurement by Meansof an Analyzer Orion EA940 Expandable Ion Method Identification Number: AMDT-M-2 Revision Number: 0 -_-- Author: Rich Youngblom Approved By: Grog] Leader Adoption Date: /0-4-45 Revision Date: None VAY 2S Date Ze AABAL Quality Assurance. Lo-#35 Date Software: MS Word 5.1a ADfofhercmtaednnDoDcXu2m0e0n0tsO:rgAanMiDcTH-alMi-d1e ATnhaleyrzmearl Extraction of Fluor-ide by Means ofa Modified O0G8Y 1 } 10SCOPE, APPLICABLECOMPOUNDS,AND MATRICES 1.1 SCOPE: This method is for the calibration and operationof an Orion EA940 Expandable Ion Analyzer. 1.2 APPLICABLE COMPOUNDS: Fluoride. 13 APPLICABLE MATRICES: Liquid samples in an appropriate `buffer solution. pHOf 6.0. 20KEYWORDS 2.1 Fluoride, fluorine, ion selective electrode Preferred 3.0 PRECA! TONS --- 3.1 No hazards identified with this`method. 4.0 SUPPLIES AND MATERIALS 4.1 '4.2 Orion Orion M9o4d0e9l9990To0t0a0l1IoenleicctSrtordeenfgitlhlinAgdjsoulsuttmieonnt(ABuCfYf)eroIrI 4.3 Orion 940907 100 ppm fluoride standard or equivalent. 4.4 4.5 Milli-QTM Magnetic water or stir bars. equivalent. 4.6 4.7 Lab tissues. Sample collection vials. 4.8 4.9 PPloalsytsitcyr1e0n0empiLpetvtoelsu.metric flasks. 4.10 Miscellaneous laboratory glassware. (TISABI equivalent. orequivalent. 50 EQUIPMENT 55..12 OOrriioonn MMooddeell E96A0994000 ESxopliadndSatabtlee`lCoonmbAinnaaltyizoenrFolruoerqiudievaelleenctt.rode or equivalent. 5.3 5.4 Magnetic Stir Plate. IBM compatible 386 or 486 computer (only neededifusing Orion 3E software). 5.5 5.6 Orion RS232 interface cable (only needed Microsoft Excel 5.0 (only neededifusing if using Orion 3E software). Orion 3E software). 6.0 INTERFERENCES 6Q6.21TMIStwaiamstprelreecwtoielmlmmpgeeenrnadetreuadrleltyhmabtarytihneegfpfseHacmtbpfelleuaottrooirdpenHemoaefra6s.6.u00r..emAen1t:.1 mixture of TISAB and sample/Milli- Itis recommended that the sample be a6t.3rTohoem traetmepethreatsuarmeplaessthaerestsatinrdraerddastwsehroeulwdhbeenctohnesimsetteenrt wwiatshctahleibrraatteedt.he. standards were stirred. 06530 - 6u.n4deArirelbeuctbrboldees. trapped under electrode can give emoneous readings. Make sure no air is trapped 7.1 No special handling necessary. 88..11.1PMreepaasruarteio5n0omfLCaolfiTbIraStAiBonITStinatnod5ar1d0s0 mL plastic volumetric flasks. 8I8a..b11e.32lePLdiapfbeleatlstkets.h0e.0f5l,as0k.s1,as0.05.,051,0,0.a1n,d0.15.,5m1.L0, aonfd110.05ppppmmfFl-u,oarildoengstwaintdhartdheindtaotethaenadpyproouprriinaitteilalys. 8.1.4 8.1.5 SAhdadkeaptphreofxliamsaktsteloymi3x0 mL the osfolMuitilolnis.-QTMwaterto each flask. 8t.h1e.f6laEslkismionvaetreathierabiurbbbulbebslefsr.om the flasks by tipping the flasks on their sidesandrolling theairin 88..11..78 IBnrvienrgt tahned vsohlaukmeetihnetfhleasfklsafsokrs up the tfointahlemi1x0i0nmg.Lmark with Milli-QTM water. 8.1.9 Record standards in Standards Log Book. 88.22.1CaIflniebcreastsiaorny, remove tape from electrode filling hole. 88..22..32 EIjnevecrtta pfreowbedrtoopwseotfftiolplisenagl.solution from bottomofelectrode to wet lower seal. 88..22..54 TFihlletmheeteelreactnrdodtehewiFt-heflielcltirngodseolaurteiotny.pically calibrated by direct measurement with no blank tchoerrmeacntuiofna,ctuusrienrg'sstiannsdtarrucdtsiownist.h concentrations of 0.05, 0.1, 0.5, 1.0, and 1.5 ppm F-, following, 8.2.6 8.2.7 RCleecaonrdthteheelselcotpreodien bthyeraipnpsrionpgrwiiatthe lMoiglbloio-kQ.TM water and wiping the sides down with lab tissues. 88..33.1SCtaolriabgreatCioonndsittanidoanrsdsfoarreSsttaonrdedaratdsroom temperature. 9.0 PROCEDURES 99..11.1CaTlhiebrsaatmipolneatnodbeMemaesausurreemdenntee,dSsttaonbdearmdixmeedthwiotdh:TISAB using the proportions 9r.e1c.2omPmlaecnedaedstbiybatrheiTn IthSeAsBammpalneufaancdtuprlearc.e the sample on the stir plate. e9l.e1c.3trAodleloiswitnhseersteadm,pmleaktoemsiuxreftohearrefaerwesneocoainrdbsubbebfloersetirapnpesd uetnhdreeertltehcietrenloedgcet.rodWeh.en the s9-a1m.e4 Trahtee assamtphelecaslhiobrualtdiobnestthaendsaarmdse.temperature as the calibratiorstandards and stirred at the 9.1.5 When the readings have stabilized, record the reading in the appropriate log book. 00591 3 9.2 Calibration And Measurement, Using Orion 3E Software: 9.2.1 Calibration: 9.2.1.1 Follow steps 8.2.1 t0 8.2.4. 99..22..11..32 TPhreesscoFmupnuctteironscKreeyen#w8il(lF8a)s.k you to confirm the number of standards to be used, Mcoankceentarnaytinoencoefstshareyscthaandnagredss,toanthdewihneftohremratoironnoptraesbelnatnekdiasntdo bcleicikncolnudCeOdNinTIthNeUcEal.ibration. 9.2.1.4 Place the electrode in the fist standard on thestrplate andclickon CONTINUE. s9t.a2b.i1l.i5zeOdb,seprrevsestAheCrCeEadPiTngsREonADthIeNgGra.phic display on the computer. When the readings have 9.2.1.6 Repeat step 9.2.1.4 and 9.2.1.5 for the remaining standards. 9.2.1.7 After the final standard, the computer will display the slope ofthe curve, as well as the intercept and correlation. Record the slope, intercept, and correlation in the appropriate log book and click on CONTINUE. The calibration data is automatically copied to C:\Orion\Data\Calib.txt. 9.2.2 Data Spreadsheet: 9.2.2.1 Select either NoE rOPW EN from the FILE menu to open a new or existing spreadsheet t9.o2s.t2o.r2eRdeactoaridn.the nameofthe spreadsheet used in the appropriate log book. 9.2.3 Fluoride Measurement: 9.2.3.1 Follow steps 9.2.1 through 9.2.4 9.23.2 Enter the nameofthe sample in the appropriate place on the screen. 9.2.3.3 Click on the NEW SAMPLE button 9.2.3.4 When the readings have stabilized, click on the RECORD button and write the result in the appropriate log book. 100VALIDATION 10.1 Quality Control: . 000000000 10.2 Precision and Accuracy 10.3 Other Validation Parameters Accordingto Reference 13.2, the rangeof detection is 0.02 ppm fluoride up to a saturated solution of fluoride. 11.0 DATAANALYSIS 11.1 Calculations None necessary. 11.2 Analyzing the Data None necessary. 12.0 ATTACHMENTS ~~ None hs 13.0 REFERENCES OGGIZ PM I1n3c.o1rOporriaotnedM,od19e9l1.EA940 Expandable fon Analyzer Instruction Manual, Orion Research 13.2 Orion Model 960900 Solid Research Incorporated, 1991. State Combination Fluoride Electrode Instruction Manual, Orion 140 REVISIONS Revision Number Reason forChange 0000 Revision Date 00693 5a Method Extraction of Fluorochemicals from Rabbit Livers SOP Identification Number: AMDT-M-4 Revision Number: 0 Adoption Date: /0-7/-55 Revision Date: None --ee---------------------------------- Author: Dave Christenson/Cynthia Weber Approved By: froup Leader Jo=3/-25 Date Gadbf Fh Quality Assurance lDeacis Software: MS Word, 6.0 `Affected Documents: M-5, Analysisof Rabbit Extractfor Fluorochemicals Using Electrospray `Mass Spectroscopy. 0594 ____ 1.1 eSEltcehocyptler:aocsTephtraiatsyemmiasestushseosddpeticostefrxootrsrctaohcpetyef.xlturaocrtioocnohfefmliucofraroolcmshetmhiecLaivlesrsfrfoormarnaablbyistislibvyers, 1L:23 AMaptprliicceasb:leRaCbobmiptoLuivnedrss.: Fluorochemicals or other fluorinated compounds, 2.1 Fcloumoprooucnhdemsi,caexltsr,acrtaibobni.t livers, electrospray mass spectrometer, fluorinated 3.1 Use gloves when handling the rabbit livers, they may contain pathogens. 4.1 4S.u1p.p1lieSsyringe, capableofmeasuring 100uL 44..11..23 EGlpopveensdorf typeordisposable pipets 44..11..45 PPllaassttiicc gcernitnrdiifnuggetutbuebses, 15 mL 44..110.67 LNiatbreolgsen 44..11..89 TFiilmteerrs, Titan nylon syringe filters, 0.2 jum. 44..11..1110 ADniaslpyotsiacballeppilpaesst:icg3lacscs vsyorliunmgeetsr.ic pipets. 4.1.12 Crimp cap autovials. 4.2 4R.e2a.g1en`AtpqspumeaoquuseAoumsmSoonluituiomnoAfceatmamteon(Aildurmicahc)e,taatpeprboyx.ad2d5i0ngp2p5m0: Pmrgepaamremoan2i5u00m 4.2.2 a`wcaetteart.eDtiolu2t1e0t0hmisLsolvuotliuomnet1r:i1c0 ffloarskaa2n5d0dipluptmetsoolvutioonl.uwmiteh Sodium carbonate/Sodium Bicarbonate Buffer (J.T. Baker), Milli-Q ab(rnNidang,21Ct.0oOv/goNloAufHmsCeoOdw,ii)tuhm0M2bii5lclaMir:-bQoWnweaatiteegrh(.N2a6.H5CgOo,fsiontdoiaum1 Lcavroblounmaetteri(cNwf,laCsOk,a)nd 44..22..43 EDtihlyulteAcaceettaotneitrile solution, dilute acetonitrile 1:1 with Milli-Q water. 44..22..65 MMieltlhia-nQowlater 44..22..78 F1CH',915H.(23HM,2SHpe-cpiearltfyluCohreomoictcaanlesDuilvifsoinoinc)acid (Aldrich) G06G8s 27 50EQ5.U1IPUMltrEa-NTuTrrax T25 Grinder for grinding liver samples. 55..32 CVoerntiefxumgiexer 55..54 ASnhaalkyetrical Evaporator 6I .0 N 6.1 T TherearE e no knR own intF erferencE es at thiR s time. E ~N ~~ CES 2.0 SAMPLEHANDLING 7.1 Tpheerfroarbmbeidt.liversare received frozen, and must be kept frozen until the extraction is 8S .0 CT A8.L1IA BPRrAepTN arIaOtiNoD nAoNf A IDnternR al StaD ndardI s ZATION 8.1.1 pPerretplauroeroaoncitnatneersnuallpshtoannidcaarcdiodftoapbperoaxdidmeadtteolyea1c2hlpipvemrs1aHm,p1lHe,.2H2H8.1.2 W1e00imghLvaotlleuamsett0r.i1cgfloafsk1.HR,e1cHo,r2dHt,h2eH-apcetrufalluwoeriogohctt.anesulphonic acid into a 88..11..43 TBorai2ni5gt0umpLtovvoollumuemwieftlhtasmrke,tiahadcndol3,mthLissiosftthheessttoocckkssttaandnadrda.arndbringto svtoalnudmaredw.ith Milli-Q water. Calculate the actual concentration of the actusaullmphgonpiecrfalcuiodroctane- X 3mL = 0IL 250ml. actual concentration, ppm 8.2 8P.r2e.p1arPereFpaCr-e9F5C-A9n5iosntanSdtaradnsdfaorrdtshe standard curve. 8.2.2 RWeeciogrhdatphperaocxtiumaaltweeliygh1t0.0 mg of FC-95into a 100mLvolumetric flask. 88..22..43DiBlruitngetuhpetosovloutliuomnewiwtihthdidliultueteacaecteotnoinittrriille1e.:10for asolution of aapsporlouxtiimoanotfealpyp1r0o0xp.pm1.0 pDiplmu.te this solution 1:10 with dilute acetonitrile for 8.2.5 5U.s0eptphme,101.p0 ppmpsmolauntdio5n000 mppabk.e working sandards with values lose 0 8.3 8P.r3e.p1arWeeiBgehef40LigovefrBoHvoimnoeglievneraitnteotao25U0semLfoNralSgteannedboatrtdlsecontaining 8.3.2 A2d00dm1LmsLMoifltlhi-eQswoaltuetri.onGrtoinad1t5mo 2Lhcoemnotgriefnuoguestsuboel.utPiroen.pare a totalofcight s1umspLenadlisqoulouttsioonf tbhye ssholauktiinogniitnbe15twmeeincaelinqfuoutgs.e tubes. Be sure to rc: 006536 F-- 8.3.3 Spike sevenofthe 1 mL aliquots with the following amountsof working standards in step 9.12 of the procedure. One 1 mL aliquot serves as the blank. Working Sandard T Approximate id (Approximate Conc.) `concentration of 7 Bm FC-95 in liver | 8.4 Calculate the actual valueofthe standards: axL.o_fsctanodanrd centat(iinoppnm) = final concentration (ppm) 171 mg liver'/ 1 ml homogenate of FC-95 in liver *Average weightofbovine liverin solution as determined by weighing F1Cm-L95hiosmroegpeonrattedesasofeq4u0ivmaglelnitvserofinF2C0-095mpLootfasMsiilulmi-sQaltw.ater. The amount of 8.5 Calibration Spectrometer. 8.5.1 Extract the spiked beef liver homogenate following 9.13to 9.23ofthis `method. Use these standards to establish yourcurve on the mass 8.5.2 Altematively, a standard curve may be generated using ratiosofresponses. of the perfluorooctansulfonate anion and the internal standard anion versus concentrationofthe perfluorooctanesulfonate anion. 8.6 Storage Conditions for Standards 8.6.1coNveewresdtapnldaasrtdiscacreenptrriefpuagreetdubweisthunetialchthaenaalnyasilsy. sSoitnsantdhaerdmsasasresspteocrterdoimneter is performed. 8.7 S8.t7o.r1ageBeeCfonldivietrihoonmsogfeonratSetsamnadyarbdesfrozen after preparation. 9.1 pOabctakiangefdrowzietnhloitvherersatmipsslueess.. In spent tissue, note that the liver has not been 9.2 Usea dissecting scalpel and cutoff approximate1l.yg of liver. 9.3 Weigh the sample directly into a tared plastic grinding tube. 99:-45 PRiuneitcdaoalrlsda.bhele olnivetrhewveiail ghtwiith de the study study noe book. number, weight, rabbi ID, date and analyst 600597 -- 99..76 AGrdidnd2.t5hemsLasmpwlaet.erP.ut the minutes, until the sample igsrainhdoermopgroebneeoiun sthseolsuatmipolnewaintdh ngroilnadrgfeorcahbuonukts.2 991.98 TRainkseetthheegprrinodbeeroafpfaritntaontdheclseaamnpiltewiwtihthme2t.h5amnLolsafwtaetreeraucshinsgamappliep.etF.ollow 59:.1110 PCAaMipDpTtl-heEmPsL-aim2np2tl.eoaan1d3`vmoLrcteenxtfroirf1u5gesetcuoben.dsL.abel the centrifuge tube withthe identical information as the grinding tube. (Sec AMDT-M-4 Worksheetfordocumenting the 9.12 Srepmiakientihnegbseteefpsl.i)ver homogenates with the appropriateamountof FC-95 standard 99/.1143 SaAspdiddkees1ctmrheiLboseafdmtpihlnee8ss.o3ad.nidubmeceafrbliovneartheo/msoogdeinuamtbeiscawribtohna1t0e0buufLfeorfainntdem1amlLstaanmdmaordn.ium 99:.1165 Uacseitnatge.an analytical pipet, add 5 mL ethyl acetate. Cap the sampalnedvortex20to 30 seconds. 99..1187 PCeuntttrhifeumgeinforthe2s0hatoke2r5fmoirn2u0temsi,nu.ntil the layers are well separated. Set the power 9.19 RmoenLmtohgvereacde4unatmtreLidfsugoglefas1ts0h2ep5tip.oept.oTrrgaannsifcelrayteherltaobealfrteosthhe15frmeLshctuebe.ntrtuibefwuitghae5 9.20 9.21 aBBrpliponrwgoxttihhmeearsteaemmlapyil3ne0idntgoows4na0mompinlnetuuhtepesai.nnal1ytmicLaldielvuatepaocretaotntoitorrinleeawridtrhyannesasnawliytthicnailtpriopgeetn., 99..2232 TVtrhoaerntsseafxmepr1l5tehseiensctaoonmadpslf.reestho cae3ntmriLfusgyertiunbgee.orAtataacuthovai0a.l2. pLambenlytlhoentmuebsehorfivlitaerl,waintdh ftihleter 99..2254 CCsotaumpdpyalnendtuhemobAledMrfDaoTnr-danMaa-nli4ymsaiwlsornbkuysmhbeeleeertc.tarnodspariayacmhatsospsapgeecotfrossctoupdyy.notebook. 10.0V10A.L1 IQDuaAliTtIy COoNntrol - not applicable 1100..32 OPrtehceirsiVoanliadnadtiAocncPuararcaym-etneortsapnpolticaapbplleicable 110DATAANALYSIS 11.1 None 1A 2.0 1T 2.1 WorT ksheet AA MDT-M-C 4 HMENTS 13.0R1E3.F1EARMEDTN-CEPE-S22 Routine Maintenance of Ulira-Turrax T-25 1Re4vi0siRonEVISIONS ~~ Number ReasonforChange. Revision Dae C0638 a EST CORY RUE AE Worksheet AMDT-M-4 Sudy 7 SNiummpbleer | apCp90r.35oppxm | apFpCIro5pooxm | approCx.555ppm| InaDlasrfeoarndtd LE ee[Ee] en[5] ma f r rt r g rF y p r t rr r r rr a r ee eerr St--] r r re eer tm mmm e r eee P r etm je] r me r et r efertreee] r St r PS SS -- r r r r r r r ttf | ER p E T -- PE -- e ----e ---- ------ -- r -- ---- Er ET eese] ETEor He e------------ T TT ees en A EErsTr ee ---- REBT ee -------------------- screams te ------ T rr ete] m rier oe r a i} [praa Bee tee] E E -- TEEree-- r r -- e-- ] mTe EE Troe raeEs e ter] C06G699 -- 3E M nvir Labo oratn oryme~n ~~tal Method Analysis of Rabbit Liver Extract for Fluorochemicals using Electrospray Mass Spectroscopy SOP Identification Number: AMDT-M- Revision Number: 0 - Adoption Date: [-( 45 Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: roup Leader 4 Quality Assurance 4 Date re Dawe Software: MS Word, 6.0 Affected Documents: M-4, Extraction of Fluorochemicals from Rabbit Livers /! [ en -- oscope 1.1 Scope: This method is for the analysis of extracts of rabbit liver or other tissues or fluids for fluorochemicals using the electrospray mass spectrometer. The analysis is performed by single ion monitoring of FC-95 anion, M/Z= 499, the internal standard M/Z = 427, and other appropriate masses. 1.2 Applicable Compounds: Fluorochemicals or other fluorinated compounds. 1.3 Matrices: Rabbit Livers (samples), Beef Liver (standards), other tissues and fluids. 20KEYWORDS 2.1 Fluorochemicals, fluorinated compounds, electrospray mass spectroscopy, mass spectrometer, rabbit livers. 3.0 PRECAUTIONS 3.1 Use caution with the voltage cableforthe probe. When the voltage cable is plugged 3.2 DinotontohterpurnotbheeDpOumNpOaTboTvOe UitC'csHapTaHciEtyPoRfO4B0E0,0tphseir.eIfisprriessksuofreelgeoctersicoavlesrh4o0c0k.0 psi stop and release pressure. The peak tubing may be plugged. Troubleshoot back to find the plug and replace the plugged tubing. See AMDT-EP-15 3.3 Do not run the pump to dryness. 4.0 SUPPLIESANDMATERIALS 4.1 Supplies 4.1.1Niogen gas regulated to 140 psi. 4.1.2 Fluofix columnorequivalent. 4.1.3 100 uLor 250 uL flat tip syringe for sample injection. ~~~ 4.2 Reagents 4.2.1 Dilute acetonitrile mobile phase, dilute acetonitrile 1:1 with Milli-Q water. 4.2.2 Milli-Q water, all water used in this method should be Milli-Q water. 50EQUIPMENT 5.1 VG Trio 2000 Electrospray Mass Spectrometer or equivalent. 5.2 ISCO Syringe Pump 5.3 Specumphysics /AS300 Autosampler 5.4 10 uL Assembly 5.5 Autovialsorcapped centrifuge tubes. 6I 0 NTERFERE~ NC ~ES 6.1 There are no known interferences at this time. 7.0 SAMPLE HANDLING _ 7.1 Keep the extracted samples in capped 15 mL centrifuge tubes or in capped autovials| until ready for analysis. 006701 5 8S .0 CAT 8L.1IBA RPrAepTN arIaOtiNoD nAoNf A CDalibrR ationD StandI ardsZATION 8.1.1 Sexetvreanctbieonefprliovceerdusrtea.nd(aSredes aAnMdDoTn-eMb-l4a,nksbeecteifolniv8e.r0)are prepared during the 8.2 8C.a2li.b1raRtuinonthe sevenbeef liver standards twice, starting with the lowest standard 8.2.2 to obtain the standard curve. Typically one standard is run after each 5 to7 samples. Choose a standard in the same range of concentration as the samples. 8.3 S8t.o3r.a1geFreCsohndstiatnidoanrdss for are Standards prepared with each analysis. Standards are stored in pceorvfeorremdedp.lasStaicmpcelnetsriafnudgesttabnedsarudnstialrethNeOanTalryesfirsigoenrattheed.mass spectometer is 8.4 S8.t4o.r1ageBeeCfonldivietrihoonmsogfeonrateBseemfayLbievefrroHzeonmaofgteernparetpeasration. 20PROCEDURE ~~~ 9.1 Initial 9.1.1 Setup Set software to "Operate on", Ion Mode ES'. 99..11..23 RFielclotrhde sboalcvkeinntgcyplriensdseurrewiinththmeobinislterpuhmaesnet.log. 9.1.4 `Sceotmtihnegpouumtpotfoth"eRtuinp"o.fStehte tphreobfel.oTwhteo p1r0e0s0suurLe/msihonu.lOdbbseeravte1s7d0r0optloe1ts800 9.1.5 pCshie.ck the fused silicaat the endof the probe. Use an eye piece to check for cchuitposf,fTthhee ttiipposfhothueldsibliecaflwaittwhitahcnoolujmanggceudtteedrgaensd, Ipfulalntyhechsiipliscasrtehrfoouugnhdto 9.1.6 Cthheeacpkpryoopurriantietrloenggetnh.supply. Tum on the nitrogen. There shouldbe no noiuttroofgetnheletiapk.ing around the tip of the probe. A fine mist should be coming 9.1.7 fCaarrtehfeurl.lCyognuniedcettthheepvroolbteagientcoabtlhee otpoetnhienpgr.oIbnes.ert it until it won't go any 9.1.8 aGpportooprtihaete"Emdaistsoer"sp0a4ge3,7anadndse4t9f9o.nization Mode to ES, and the 99:.11..910SI1fairtitsAncoqtuiinsisionng.leAsfosnigmnod& eigonatome",OpMtOio-nD"AaYnd-sYeRt S+IRe.ter. Record tin 9-1. 1Rtuhenlotgheboboeke.f iver samplesfirst running each standard tics at the `5b1e0gi7nnsianmgpolfest.he run... Run a QC check by running one standard after every 9.2 9M.a2n.u1alDrIanwje1c5t0iounL. of sample into a syringe. Injec-t the sample into the rheodyne: 9.2.2 Tinujencttihoenvpaolrv.e I1n0je"cOtn"s.lowly. Record the sample ID in the log book. 99..22..3 RWeaciotrtdwothmeisncuatnesn,uamnbderinfjoerctetahcehnseaxmtpslaempilnet.he logbook. - C06 702 gos -- 9.3 9U.s3i.n1g StehteupAustaomsplaemptlraeyrA, B, or C. 9.3.2 Rvieaclosrmdatyhebesaimnpelaecsharnudn.their positions in the instrument log book. Up to 17 9.3.3 9S.et3-.u3p.t1he samplPeurs:h the samplebution 99..33..33..32 SSeetrisnajmeplcetslaomopplsei=2z=e 100ul 99..3333.154 SNeatmCeytchleeftilie:meLi=v0ers 99..33.33.76 AIddedntitfhye sthaemptlraeysutsoeQdueuebypressing "Enter" 9.3.3.8 Press "Run" to start 10.1 Q10u.a1l.itTyRuCnonatsrtoalndard every 5to 7 samples.If a significant change( 50%) in spteaankdahredigwhitllocbeeurussesdi.oTp hteherreumna.iOnnilngy stahempslaemsplweisll bbeeforreeantahleylzaesdt.acceptable 10.2 P1r0e.c2i.s1iSoeneaMnedthoAdccVualriadcaytion Report number AMDT-M-5.0.V1 1100..43 ROetfheerrtoVMaeltihdaotdiVoanliPdaatriaomneRteeprorst Number AMDT-M-5.0.V1 1111.:12 PClaolcrutlhaetisotnasndard curve, using the meanofthe two values obtained for cach s1t1a.n2da.r1d.Read peak heightsorareas for thesamplesfrom the printout. Use linear 11.2.2Craelgcrueslsaitoen to the determine the mg of FC-95 sample concentrations. anion, or other fluorochemical in the total rabbit liver: mg FC-95 anion in the total rabbit liver = mggmEsCo-f05liavneirounsfedrofomrsatnda.lcyusirsve ~~ x Total mass of liver, gms 1111:4.3 PMrainktea acrhersoulmtasttoagbrleamanfdoreentaecrh ist aimnptlhee,swtiutdhy book. the peaks labeled with the sample or ssttaunddyafrodldIeDr.. WSrtiaptleetahlel scthurdoymnautmogbrearmosntothgeeptrhienrtoanntd,niunimtibael,rdpaatgee,sa.nd put it in the 606703 yo None 13.1 AMDT-EP-17 140 REVISIONS Revision Number Reafos rchoannge ---------- Rosin 600704 se 3M Environmental Laboratory Method Analysis of Fluoride UsiTnogn tSheelecStkiavlearElSeecgtmreondteed Flow Analyzer With Method Identification Number: AMDT-M-8 Adoption Date: 0-5-3 Revision Number: 0 Revision Date: None e --e ---------------------------------- Author: Deb Wright/ Cynthia Weber Approved By: up Leader 4 a4 2 Quality Assurance o/s7s Date J-2 7-45 Dae SAoffftewcatreed:DoIcBuMmeMnStsW:orAdM,DT6.-0EP-26, Operation and Maintenance ofthe Skalar Segmented Flow Analyzer [FS 1. woscoee 00000000000 1.1 This method is for the analysis for fluoride, thermally extracted from samples using the Dohrmann DX2000 (AMDT-M:-1), and collected in TISAB for analysis with an ISoengmSeelnetcetdivFelEolwecAtnraoldeyz(eIrSEw)i.tThhIeSEa.nalysis is performed using the Skalar 1.2 Samples can be tissues, serum, biological material,or other materials extracted on the Dohrmann. 20KEYWoRD 000S 00 2.1 Skalar, segmented flow, fluoride. 3.0 PRECAUTIONS 3.1 Follow standard laboratory safety practices. 4.0 SUPPLIESANDMATERIALS ~~ 4.1 4S.u1p.p1liSeasmple cups, 4mL plastic cups with caps. 4.1.2 Autopipets, oxfordorequivalent with plastic 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 nSuammbpelresp.refilters, Evergreen 4.2 Reagents 4.2.1 Brij 35, 30% S.F.A.S. Detergent 4.2.2 TTIISSAABB IIII baudfdfe2r.5smolLutoiorn:1P0u0rpchpamseflTuIorSiAdeBsIoIluftrioomn Oarnidon1.mTLo B1lriitj.er of 4.2.3Samplerrinsing 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 mtLhesvoclonucmeenturpatteod1nLitrwiictahciMdil(lHiN-QO,w)atteor.250 mLs of Milli-Q water. Bring 4.3 S4.t3a.n1daSrtdosck solution, 100 ppm F: purchased from Orion. 4.3.2 Intermediate standard, 10 ppm: Dilute 10 mLs of stock solution to 100 mLs 4.3.3 `with Milli-Q water. Use polypropylene volumetric Working standard: Make up the following working flasks. standards by adding the `vooxlfuormdesoropfuimntpemramtedeiaptiepeosr,sttooc5k0smtaLnsdoarfdTiIndSiAcBateadnodnditlhuetitnabglet,o u1s0i0ngmLs with Milli-Q water. FM ooopsem em-------- o0r3s || Ooe memn-- >| 15 | opm[0 | 3 G0C 706 2-- CC Teppm C--tsepm 1 12 --T ----- 1 |ts] 50EQ5.U1IPSkMalEarNSTegmented Flow Auto Analyzer SansTM System equipped with ISE 6I .0 N 6.1 T High coE ncentratR ionsofaF lkalinitE y, chlorR ide, phoE sphate,N sulfateC or ironE can cauS se interferences. 7.1 `Swaimtphilne3s0shdoauysl.d be stored in polyethylene bottles. Samples should be analyzed 8.1 P8r.e1p.a1raPtriepoanreocfalCiablriatbiroantsitoanndSatrdasndaasridnssection 4.3. 8.2 8Ca.l2i.b1raTthieonstandards are analyzed at the beginning of the run. 8.3 8S.t3o.r1a"geSSttaaCnnoddnaadrrdidsstairaoernepsrsetfoproaerrdeiSdntacatanapdpmaeriddnspiomluypmorofpyelveenreyvsoilxummoenttrhisc,floarsakss.nNeceewssary. 9.0 PR9.O1CEStDarUtRUEp Procedure 5.1.1 Clamp down the pumpdecks,si bas nd sampler pump tubing. 99..11..23 TPuutmthoenfltuhoerpidoeweelrecotfrotdheessianmptlheere,lepcutmrposd,e cohffasmebterp.otentiometer and heating. 99..11..54 TbPauutthmt,hoenretahgeeinnttleirnfeascei,nctohmepauptperro,prdiiastpelaboyttalneds.printer. Make sure you 9.1.6 tLeutrntheonsysttheemisnttabeirlfiazceeforbeafpoprreoxitmhaetecloym3p0utmeirnu.tes. 9:2 9S.t2a.r1tinagCreRateuansample table by sclecting FILES, TABLE, and CREATE, type in 9.2.2 Pthreinntatmheesoafmtphleeftialeb,lea,nidnsperretsesdEiNnTthEeRs.ystem table by pushing ESC, 9.2.3 DPiRaIlNtTh,e seconds GsaRmpOlUerPsc1.itTihnigss twoiltlhepraipnptrtohperieanttierneurmubn.erofsamples, number for sample wash, and numberof seconds for the sample. of 9.2.4 FFiWll/RhUeNOsaUmTplceumpayswointhtthehesasmaplnedredodnsoatmpnleeesd,twoabsehefislleadn.d drifts. TW and 9.2.5 Set the baseline. 606707 3-- 9.2.5. m1Sacylebcet iGnRtAhePHDaItCaSH,anRdEliAnLgTPaInMeEl..SIwfiytocuhctaontnhoetAgneatlyrseails-Ptiamnee,lyboyu 9.2.5. Sb2eaUlsseecetliitnnhege.CsAmdOajlNulsTtsRctrOheeLwbdarPsiAevleNirnEefLournattnihdleipotfuifsssheaitpnppgrootFxe7in.mtaitoemleyte3r/t4oisnecthtfherom 9.2.5. t3hCehbeoctktotmheohfitghheesstcrseteann.dard and adjust the gain, if necessary, with 9.2.6 GCoAltao0CttObhNee TirunRnt.eOr(fLSaecPeleAscNctrEoeLrw,d#ea3s.neldetcotaannaallyyssiisspbayneplr.eDsessinegleEcNtTtEheR.an)alPyrseisssthTaatb 9.2.7 92.8 PtorerseswthmetsoptahceebAanraltyosbirsiPnagneulp. the local menu. Select STAtoRstaTrt the anaslaymspisl.e table which you created under 9.2.1 (er 3.3.5 T$y2p8e EyoNuTrEIRD f(ionritcihaolis)c,est)h,echoose running with or without the system table 9.2.10asAnfttdieorstelhsetecattTSiignThgtAtnRhueTmsbAoefNrwAaoLrfYes,SaIsmtSpal.rtesthaensdamtphaltert.heMsaakmpelesu/rweasthh/aatitrhetismaemsplareer is 9.2.11 SOeKle.ct GRAPHICS, REAL TIME to view the progress of the analysis. 9.3 L5oa3d9inSgoo'atneoedlCeOcPtrNLiTnOtRAiODn.Lg SPtehAleeNcEtDLaA,tUap-TrFOeisClseAtLheCsUpaLcAebTaIrOtoNbarnidngeautperthtehelofciallemnaemney(or 9.3.2hTigohvliigehwthtehecfaillibfroatbieonprciunrtveed, agnodtporGesRsAEPNHTIECRS),.CALIBRATION 99.:33.43 ToURprViEnt.the To print the lhoigwhlleevveellsccurreveen,, Change the max ppyuuvssahhluEPeSRtCIoNtaToppgSreCotxRoiEumtEatoNef.lgyra9p0h0i.csG.oSetloecCt,AL 9.3.5 `CpRrSeUetEsRuTsrTVnEItENNoGTaSSnE.EdRLpTraenIsdNsGPESRSICaN,nTdacnShdCaERnngEteeErtN.hePtromeapsrsxinPvtaRslIuaNemTpblaSecCkpRetaEokE4sN.09.5, go to EDIT, 9.3.6 TOoUTpPriUntT.theSerelseuclttsPRgoINtoTCEORNfoTrRtOheLEpPsAoNnEoLr,PSRPNACfoErBtAheRL,asOerU.TPUT, 9.4 99S5..h:44u4..t312doPLKuewettenatrlhlheteshyessryteseatmgeernmitnh-slaeisnfreoisrnisanepdMpircloolxmiip-mlQaettweealltyye,r3.t0umminouftfetsh.e sampler, pump anqduid in fsa potentiometer. Tom off the heating bath on weekends. Leave li 99.744.745 TtRheaeellkeeelcaitnsteehFse.IteLhlEeeSpc,utrmpoprde-esdsoeucAtkLsa,TndaFisraobnaadkrsisnealne1dc0t0sQapUmppImlTFertoopveuexmrtnpi-tgthhuetb.ipnrgo.gram. 95.:44..6 SOn Friday, tur off the computer, display and interface for the weekend. 10.0 V1A0.L1 IQDDuaAaliTtYIySiugOCnnoiNnfaitcsratonatlncdhaardng(emiidn ptoeahkighheicgohntceonctcruartsi,=oon)nleyvetrhey s1a0mpslamepslbeesf.orIefathe last Septable standard will be used. The remaining samples will be reanalyzed. 606708 - 4. 10.2 P1r0e.c2i.s1iSoeneaMnedthoAdccVualriadcaytion Report number AMDT-M-3.0.V1 1100..43 ORetfheerrtoVMaeltidhaotdioVanliPdaatriaomneRteeprosrt Number AMDT-M-8.0.V1 11.0 D1A1.T1 ACAalNcuAlaLtiYonSsIS T1. 1. 1The standard curve is plotted by the Skalar software. 11. 1.2 Ablelterc.alculations are done by the Skalar software. * should be 0.995 or 1111..32 WPrreiptaertehespsrteuaddyshneuemtbsetroosnumtmhaerpirzinetoduattsa,. iInnictilauld,edastaempthleepvroilntuomuet,,awnedibghitnsd utsoegdetehtec.r Swiutmhmaallrypaschkeaegteanddoctuampeenitnstoatnhde psltaucdeyinnottheebosotku.dyBfaocldkeru.pMalalkdeataacaonpdysopfretahdesheets 11.4 Eo1ln1et.co4t.rsot1nuiGdcyLDPdaitssatkudaineds:bEalcekcutprodniiscksd.ata is copied onto the Study floppy disk for cach 11..4.2s0ttuhdey,r satnuddiaelss:oAdlaltdaaitsaciospcioepdioendtoonatofalofplpoyppdiysdkisthkatthiastsitsosrteodreidn tinhethleabl.ab. 1A 2.0 TTACHMENTS None 13.0R13E.F1 EARMEDTN-MC-E1,SThermal Extraction of Fluoride by Meansof a Modified Dohrmann 1133..23 SDAkMXaD2lTa0r-0EM0ePOt-rh2go6ad,nsi,Ocp#He3ar3la5it,dieoSnkAaanlanaldryMzMeaerit-nhLtoiedvnesarnMcaenuofalthe Skalar Segmented Flow Analyzer 140REVISIONS Revision Number Reasonfor change Revision Dae 000709 Se 3M EnvironmentalLaboratory } Method Thermal Extraction of Fluoride by Means ofa Organic Halide Analyzer Modified - Serum Dohrmann DX2000 Method Identification Number: AMDT-M-14 Revision Number: 0 Adoption Date: 0-375" Revision Date: None Author: Rich Youngblom Approved by: En) ip Leader Jo/5/ 2S Date Lhe AL Quality Assurance Gmzpi-- Date Software: MS Word 5.18 Affected Documents: AMDT-M-2 Fluoride Measuremenbty Means ofan Orion EA940 AMDT`EExPp-an3daRboluetTionneAMnaailnytzeenrance Organic Halide Analyzer ofa Modified Dohrmann | DX2000 000710 r= 1.0 SCOPE, APPLICABLE COMPOUNDS,ANDMATRICES a1.o1nSicdoeper:oTmhivsarmieotushomdaitricfeosr.thTehoepefrlautoiroindeoisfatyDpiocharllmyanconllDeXct2e0d0i0n TwIhSeAnBitsiosluutsieodntfoorexatnraalcytsis w11i32thMAapatpnrlfiioccneassbe:lleeBciCtoiolvomegpieoclaeuclntrdfolsdu:eid.sF,lpuaorrtoiccuhleamrilycaslesruomr.other fluorinated compounds. 2.0 KEYWORDS 2.1 Fluoride, fluorine, extraction, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, DX2000, fluorochemicals. 30PRECAUTIONS 333..213 GPGrlleaassssssuwwraairrzeeedaisngadfsraeegsxi,hlaep,ursobtpreogrkasecenosmgpclaarsnessbsmeeadeyxgtcaraseumhseaelnyidnljhiuonrtgi.esp.ractices required. 33..54 PSootlevnetnitalbbaisoehdaszaarmdpsledsuemtaoythfelabsih,olmoagiycanlemeadtrtiocaesl.loUwstehaepmprtoopdrriyatdeopwenrsboenfaolrperosttaercttiinvgerun. equipment. 4.0_SUPPLIES AND MATERIALS 44.21 43 CCoommpprreesssseedd Quartz glass HOexlyiguemn,, HHiygdhroPcuarirtbyonGrfardeee,, rreegguullaatteedd ttoo 3405PPSSLL. sample boat with TeflonTM tubing, Dohrmann 890-097 or equivalent. 44.45 OQurairotnz9g4l0a9ss99coTmobtuasltTioonnictuSbter,enRgetlhiaAndcjeuGsltamsesntGB-u9f4f0e5r-(0T1I2SoArBeqIu)ivoalreenqtu.ivalent. 44..76 SMialmlpil-eQcTMolwlaetcteiron vials, HDPE. 4.8 4.9 APocltyisvtaytreednCehapirpceotatels,. E. Merck 2005 or equivalent. 44..1110 MHiasmcielltloanneSoyursinlgaeboorrateoqruyivgalleansts.ware 50EQUIPMENT 55.21 RIoBsMemcooumpnattiDbolher3m8a6nnorD4X3260c0o0mpOurtgearn.ic Halide Analyzer, modified for fluoride extraction. 53 5.4 DExXc2e0l 0S0prseoafdtswhaereet,, vveerrssiioonn 15..000,ormogrdeiaftieerd for fluoride extraction. 6.0 INTERFERENCES 6.1 Sample size is limited to approximately 100 wl. This may vary from matrix to matrix 000711 2 o==e=. PLE HANDLI 2.1 Samples ae to be handled with plasic pipettes. A new pipet is to be used for cach sample. RATI D I 8.1 Preparation of Calibration Standards 8.1.1 The standards required for cach project will need to be appropriate for that individual project. R8e.1f.e2rTtyoppircoatlolcyol50f-or50th0atppprmojFecCt-.95inmethanol standards are used. 8.1.3 For rabbit serum studies, use. `beef serum as the matrix. 8.2 The Calibration - Overview normal calibration is the fluoride curve (AMDT-M-2). However, ifan optional spiked serum curve is required the procedure listed below is used. 82.1 A calibrat and combusting ion curve fo them using r th the e DX2000 is generated by same methods and matrix spiking samples wi type as the `samples th to known st be tested. and ard s 8s.p2i.k2edT.ypically, Ps Standard three curve replicates. ofeach standard and five concentrationsof will beisp.lotGeeenderasatMeaassreSgrpeiskseidonFc(uurgv)eoanndthcealxc-ualxaitse standards wi atnhde eSqtuaantdiaornd ll be oMrastsh line Bor vered F (ug) on the y-ax and the 12 value. 8.2.4 Mass Spiked F (ug) = (Amount spiked in mL) x Conc. ofstandard in ppm) x (0.6004)* 8.2.5 *FC-95 is 60.04%F Standard Mass Recovered F (tuhge)re=fo(rTeI0S.A6B00v4oilsu`mthee ifnacmtoLr)usxe(dOtroiocnonrveeardtiFngC-i9n 5pptom)F 83 Calibration - Procedure 83be33.1u1.se1dSitRfaurttnheU2pporrevmioorues CsalmepalneCsyccolnetsawinheednhistgahrtcionngceinntsrtartuimoennstoefacflhuodraiyd.e.More clean cycles may 88.332.2.1BlParnepkasre sample using the same methods and typeofmatrix as the test sample. 88.33..32..32 analyze sFDuoatrmpsrlaeebrbuaimctcsbotlruaddniikensgi,ntuoDsmeoehbtrehmeofadnsnAerMbuoDmaTta.-sMtC-hoe2mmbafutosrrittxhs.eafmopnlesealsecdteisvecreilbeecdtriondseeacntailoynsi9s..0 and 53.0.4 For rabbit studies, the meter reading n. Bum foar blank sample should be 0.03 ppm or lower samples until this limit is reached, or until in the jbuefdogreempernotcoefedtihnegopweirtahttohretchaelriberaadtiinog is stable with respect to historical readings (previous 48 hours). 8b.e3f.o2r.e5Fproorceneodni-nrgabwbiitth stthuediceasl,ibtrhaetibolna.nk readings should reach a predetermined ionconcentration of charcoal with the sample to aid 83.2.6 It may be necessary to mix approximately 50 mg - `combustion. 000712 , ---=, 88.333.3.1SItfabnedcasredrCuumrvisefrozen, thaw at least enough to complete the standard curve analysis for the day (=30 mL), B$33332 SPuipPeCtVteih10t0heu Lloofwbesetesftasnedraurmd cionntoceDntorhartimoann.n Ussaimpnlgae Hboaamti.lton sy ring e, eject f ixed ect i t on oqouraindntitthyebAoetfeltfhaessesrt3uarnmed.palridcaotneosrshionulthdebmeatursiexd. fFororthreablboiwtessttudsiteas,nduasred4counLceonftsrtaatniodna;rmdoarnedreejplicates `Emayebe 333% uCsoemdbautstthe.thdeisscaremtpiloen Rumall 15 standards. oafsftodhenesecarnriaeblpeyldsitci.anteseisctsiiognni9f.i3caanntldy adniaflfyerzeen tacfcroormditnhge toot hAeMrDtiTv-oM-2. es the. `roelpdlirceaptleisc,atreunanadnotthahterthseanmpelwerfeoprlitchaattnesrwtuianln,ldacbraedl.cuusleIandtdeitcotahcteaelcri2un.ldaat2teamtthuhesarttebtghereeasntseilwoenarscetup0rl.vi9ce5a.t.e replac Ifit is mot at l$ae3a.sbt307.9W5A,hmceoonnwsaulsllttaaswnntddiaaantrrdhaddsrccudpuuserrrhvvvaeeivsmseoharbo.yueleadlsboebreurnuwnhaetnthtehdeicscormebtuisontiooftnhteubaenoalryssatmple matrix is Changed. New st 8.4 Storage Conditions for Standards T8.y4p.i1caSltloyr,asgteanrdeaqrudisreamreenstosrfeodr asttarnodoarmdsteamrpeedraetpuerenidneponlnatsttihcesicnrdeiwvi(doupalbosrtaensd.ards used. 8.4.2 New FC-95 standards shouldbeprepared at least once a`month. 9-- 0 PROCE-- DURES ---------- 999...111.2T1yOCpoximycbgaulesnOtpiaeonrndaHttueiblneigutCemomnfpdleirotawit=uorn5es0:=ce9/5m0inCu.te. 91:3 Vaporization/Drying time = 240 seconds. 9.1.4 Bake time = 300 seconds. 99.22.1StafrtthUe pprPorgorcaemduisrneo:t stated, sart the EOX program on the PC. 9.2.2 9.2.3 Open the SYSTEM SETUP window. Put the fumace module and the cell in the READY mode. 9.2.4 9.2.5 Close When the the SYSTEM SETUP window. oven has reached the READY temperature, run the CLEAN BOAT program found 9.2.6 in the CELL CHECK menu. See AMDT-EP-3 for details ofthe Dohrmann software. 9.3 Sample Extraction Procedure: 9.3.1 Open the SAMPLE HATCH and pipette - 1001 of sample Tito the BOAT. It may be `Gnoencee,sscahrayrctooamlisxhoauplpdroaxlismoabteelmyix5e0dmign wohficlheaersctoaablliwsihtihngthtehesabmaspelleinteo aid and cwohmebnusgteinoenr.atIifntghitsheis standard curve. - 9.3.2 Close SAMPLE HATCH. 000713 = 9S .3.3 AD dd approHpericaiteovnolvuilm.eoTfypTiIcaSlAlBy 0so6lumtiLontoor131:m1iT.IaSreABuIeMd.IIFloErQrnabwbaittesrtumdiixeis,uruese013.0 or 20 mL Be oGfas1:eh1seTrIevlSiaeAlaBss:eodMhidalutrliihn-geQpTMtyiprwoaoltfyestrihsemimCxutOsuMtreBb.uUbSblTeItOhNrouTgUhBtEhe iTIiSnAtBhe. TISAB at least 0.25 923.5 oS nmRuenenteheedEBOOXTXI-SWpAArBoTgwEraaRsmpusrsoeAgdnritasomhceofdlo,lurcntedmtiohnve teshatemhRpelUceo,Nllaemdcedtniaounn. evqiaulalrvoomluthmeeocofmMbiusltli-oQn nubwea.ter rrtoeemthoeviTemIaeSnnAyBtThIetSoeAnmdBakorfetehm1ae:i1ncTioInmSgbAuoBsn:tMtiihoelnltfiuub-beQe.TMnw.iytrheMmaiilnli-ngQTMashwafteormanthdewbiopate.wiAtshhacKenIMbeWfIePmEoyteod sODmOoepvathpeaesladmapalpeoprlhiatcthactehorrasanindmdirlloearmrovdveaveciucauem.edAodur.opoItfmMaiylblein-eQcewsastaerry 0masycubpe padadreidcetso otfhfe the `99b.3o3.a.t11t10o iCRlduonsinetththeheeCChlLaetEcahdnA.yNCfyocBrlOea.nAaTlyspirsobgyraomn. selective electrode (AMDT-M-2). 9312 Sample is rea 9.4 Sample Calculations 59.:44.13 USsoemthee sMtaasndsaRrdeccouvrevreetdo cFal(cuugl)a=te(tTheISsaAmBplveolvianlume.L) x(Orion readin(gSlionppep)m - nterespr) 10.0 VALIDATION AI 10m.a1liQeu3aJlsaiatrySttCeaodrnttberUydopsalanCmahplylezecisknugSsaQemdCptlsoeagsme:pnleOernsa.cteeTtthhhee sQsttCaanndsdaaarmrddplcceuursrvvaee,rsEtaeoscbtheabctlohineschseenadtm,reactaaiscohtnhdmeauylsootwfebsot iomnonstpiuknlos hefirst two replicates are within 20%of th standard curve, thena third replicate is not necessary. L1O0.2RPreeciesiomn aand 2A.c3cuarnadcy5.:SPreeecimsieotnhoadnddaecvceulroapcmyenvatriaensalwyhsiesnaanndalsyazmipnlgesaanmapllyessiosfindifferent `matrices and different reference compounds. 10.3 Other Validation Parameters: NA 11.0 DATA ANALYSIS 11.1 Calculations = 1111.11.21 TFoorctahlecusltaatnedatrhed fcluurovrei,deusceonrtergarcetsisoinoninatnhaelyssaimspilne,Exsceeel,mevetrhsoidonA5M.D0To-rMgr-e2at.er. 000714 = 11.2 Analyzing the Data e112t.1hi2nm2u0s%toobffetehaaetclahevaesorttah0ge.er9o5afnodrthtgohrseeeaottuewtrlo.i.e"rOAuintslygireseruastc"ehrmotauhtyalnieb2res0es0xh%cooluuflddethdbieefapvoteiwrnotaegodef ootuhftethtinhorsteeheetrwdeapotloiacraatneds Hnoetehdainnt5he0F%inal Report along with the reason it was considered an outlier. None 13.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer Operator's Manual (Manual 915- 31439.,3 rAevMiDsiTo-n MB,-D2ecFelumobreirde1M99e3a)surement by Meansofan Orion EA940 Expandable Ion Analyzer 133 AMDT-EP-3 Routine Maintenance ofa Modified Dohrmann DX2000 Organic Halide Analyzer 140REVISIONS Revision Number `ReasonforChange Revision Date 000715 6 =, 9.1.3 Amendmentto Analytical Protocol AMDT-020795.1 000716 ~--- Attachment [ GLP Study Protocol Amendment Study Number: AMDT-020795.1 Study Title: Single-Dose Dermal Absorption /Toxicity StudyofT-6052 in Rabbits Study Director: James D. Johnson AA mendmembaue NovewberT > > T Amendment Date: November 8, 1995 `Amendment Number: 1 This amendment modifies the following portion of the protocol: AMDT-M-14-0 specifies using bovine serum for the blanks in the thermal extraction. However, rbalbabniktssweerruemfhrigohmetrhtehacnotnhtreoslaamnpliemsa.ls (AMDT-110394.1) was used because the bovine serum. Approved by: dy Director, 1 go 75 ate - 000717 5. 9.3 Quality Assurance Unit Statement 000718 Anachment D GLP Study Quality AssuranceStatement Study Title: Single-dosTe-6D0e5r2mailn RAabbsobripttsion/Toxicity Study of Study Number: AMDT-020795.1 Name of Auditor: Kari Rambo TThhiesfsitnuddiynghsaswebreeenreipnosrpteecdtetdo bthyetshteuQduyadliirteyctAosrsaunrdanmcaenUangietmea5nti.ndicated in the following table. Inspection Dates From To 10/1395 10/1995 Phase Final Report Date Inspection Reported to `Management StudyDirector 10/1955 10/1995 A QAU , Aud y itor y 10-D0a3e-35 000719~-- 1 9.4 Key Personnel Involved in the Study 000720 * 3MEnvLai borr atoo rynmental Key Personnel - Thermal extraction followed by analysis using Orion ion analyzer: JDiembJWorhingshotn Rich Youngblom Deann Plummer . Analysisofliver extracts using electrospray mass spectrometry: Jim Johnson Dave Christenson Tanhaelrymzaelr weixtthralcotniosnelfeocltlivoeweedlebcytraondael:ysis using Skalar segmented flow Jim Johnson Deb Wright : + Rich Youngblom Deann Plummer Documentation and Reporting: Jim Johnson Rich Youngblom Quality Assurance Unit: Gale Van Buskirk Cynthia Weber Kari Rambo 000721... 9.11 Data 000722 9.11.1 Summary and raw data; ug F~ in whole liver as determined by thermal extraction followed by analysis using Orion ion analyzer. 000723 ___ Summary of Combustion Data - Liver AMDT-020795.1, BWI 6329-135 As Referenced in Final Report section 6.0 DATA ANALYSIS Total ug Fluoride in Whole Liver Mean per Dose Group** ug Std. Dev. Control Group 16.7 + 5.9 2.0 mg/kg dose (T6052) 12.6 + 32 200 mg/kg dose (T6052) 18.3% + 4.2 1000 mg/kg dose (T6052) 24.1 + 42 a~OCnaelcouultaliteerdoamsitthteedm.eaVnaolufetroifplriec-aatnealsyazmepdlsesamfprloemienacclhudoefditnhrteheismarleepoartn.d three female rabbits. 000724 _. FC120 48 pAoctmuFal- ApvpermaFg-e liver Wihvoelre TotwahlolFein i % nliver ory WW) inliver WW) bumed weight rams) Gams) liver G0) Dosage (Moko) LLiivveerr BBllkk-:12 00..113038 00.112162 LLiivveerrSSppk-k12 8900%% 01..91800 00.113190 LLiivveerr SSppikk-e3d Lverspks 9924%% 21.7026 108% 324 00..110352 0.101 LLiivveerr bSlpaknck6-3 94% 0234833 00111197 Liver blank.4 FF5s22007722-12 00332600 00112017 825 037 031 0103 85 203 00 F520723 FF5522007733-12 00321450 0o1u3s2 785285 0243 0251 0150 758 190 00 F52673.3 FF5s22667759-12 00226194 00113119 795088 0171 0183 0148 908 168 00 FLi5v2er07Bl9a3nk-1 00.217688 00113134 908 Liver Blank.2 LiverSplke-1 80% 00.183614 00.113415 LLiivveerrSSppilkkee-23 8799%% 01..83653 00..110308 LLiivveerr BSpliaknek-4A Liver Spikes 77% 88% 00.090605 1.02 00.113221 0.130 LUivveerrSSppiikkee-76 9902%% 01..90943 F52675-1 065 00..113348 0136 864 F520752 F52075.3 0011138 018 0011335 886644 120 00 F52076-1 Fs20762 001132s5 0185 00114510 66088 112 00 FFe522008736-31 F52083.2 00210543 00112374 690685 01st 0s Of 865 140 00 FFe522998863-13 Fs2088.2 00.113429 00112285 988258 ow omy ots 828 122 20 FF5s22000806-.13 F520002 00..114652 001134s5 882084 0196 018 0120 804 148 20 FFs522000070.31 F520072 00129250 00110174 890147 0263 0195 oie 917 178 20 F52907.3 0123 0150 917 RPTIISLXLS Page 1 000725 RPTIISLXLS Fct20AB pAoctmuFal- ApvemrPag-e liver Wvhoelre TotwahloFl-ein % iver iniver bumed weigh Iver Dosage FFss22e0882242 vy W01W3 ) 0153 (WW) 013 (0gr1a4m7s) (g6r2am4s) 0105 624 (u)) 10 (mghkg) 20 FsF2s029040-213 Fs20042 00112113 o0t1e0 682249 ois oa 01st &29 93 20 FsF2s04910443 Fs4102 o0t1t13 0o1i4s8 2eto9 0130 0115 0419 910 105 20 FLi5v3e4B1l0a3nk-1 00210072 o01u0s8 90 LLiivveerrSBpliaknek-2t 65% 00911310 00110082 LLiievreSrpsipkiek2e 7615%% 0O7A5T1 00113a1 LLiveerrSsppiktee4s 9865%% 111032 0011225s LLivveerrsSppiikkee78 8900%% 0190448 00113218 FsF522000844-21 0022146 020 oOi014z s8e5s 177 20 FsP22008024:3 Fs2s022 0016703 00112311 8a8oss 0s 029 Oif5 08 168 20 FFss22s00062-31 Fs2006.2 00828437 ooi t 1068 022 106 ois Ts 825 200 re"aSnaamlpylzeed FFss22009767..31 Fs2077-2 0220077 ootz 712708 below 010 0187 01s 720 135 200 FF5s22008707--13 P2082 00211773 ooiis 977209 026 0219 014 oT9 25 20 FPs220000043 Fs20932 00119724 0o1t3es 808793 0199 0175 0129 883 155 200 Fsv2er09bl3a3nket oa01r523 ootee 683 iieerrssppiikkeet2 7725%% 00794218 iversplked 67% 0870 00.114283 0151 iIvveerrssppiikkees 7777%% 00883248 iverspike 88% 127 00114216 010s -. Iivveerrssppikkee? 18512%% 0185229 00115418 iivveerrssppikiee-d10 8808%% 11.3122 0010118 Page? 000726 RPTISSLXLS Fo120 a8 mActuFal Aovemraege Iver Wvhoelre Totwalidein D Fs2008-1 % inliver iy (0W20W1) inliver WW) bumed weight (EgemsE) (Egrams) liver (uo) Dosage (moka) pages FsF2z9e0o6s.23 Fazeret 0042242 0258 0319 0otMo om 7T778s wz 248v 20 onfaablovse FFaez2eu778832 Blank ver 1 00222254 oze ooimla 8m22 198 fom oatt 0120 BBllaankkIvveerr 23 "20a1s BlankIver 4 0340 0o0t14i3s7 BBllaannkkIIvveerr 6 Cverspket 00.241074 130% 1.37 00114154 014 ULivveerrSsppiee2 Liver Sed 8o47%% 62% 0180391 0.955 0o111s7s 010 FsL2v0er8s0p1kes Ti 00728848 Fa20802 Ops ozs o01t5s0 ous 6s2o0 27 1000 FsF2ao200810-.13 Fazoo12 00233%1 G20 oz ooflsss 017 s8e2s0 ss 243 100 FaF2s209007113 Fazoer2 002204 0o2i0 8s1e2s O04 Os 0d s1z 20 1000 FaFsz2o9e87s.13 Fezoss2 00.22360 Gio om ooiilss orm 88122 m2 29 1000 FsFsz2e0s05e.s1 Fizo0s2 00435430 031s ous ootro oi e8a3t2 1 28 1000 Fsvze0r0b5l.a3nk 0ol2e6s8 Iverspkod 77% 0.862 o01i4s6 eat 0128 iIvveerrssppihkoes2 8812%% 008.2407 Iverspked TT 0.905 0o1t0e 0129 IIvveerrsspplkkeesd Iverspket 8643%% 210889 81% 175 0011325 0141 ver bank 026 oat Pa"ges 000727 -- 9.11.2 Summary and raw data; analysis ofliver extracts using electrospray mass spectrometry. 000728 -- HW! #6329-135 + WPCaolnemsiegnggpay < hs sTEorreo :tuam: wRnRessis:oende Fvacato:r Arun: oA tros: Caoramseen:a: SThoistooichaotin(Gs.o1r2pt0ion fLNooxer aBos pFrien Ya200 cronUS Pe arson Lat Shon achenchsstreenad fn A ST Bn aony,ThoIgoandiUiabibon periemo9, Amt gwen gg ob:er Fodmpriewe dsof [BRiamams waeaaa a FRaamn N"NNDoD. oomaoha =| Fris nNoo.. FRRaarme o"NvNDGa Hie Ne mwoTra+| rFammse 3 F[ Bais Awaobm Fm ND, a onor=a | rBawms JJRee S Fi moma Si Mabo5ragn 5o5. [I---- = ists a o vlana of 20 mA = Kimoso vonr o001a mh - TaTccolonocraonnrteaiatnnotwfe8sacnaacl7kc1ni7io5oynTishngo g sa rodmardn5i54vnsrWaadtnhlaosiHnTg hr0e.rso1r3y7s1 45 rTohnr41 aa5saryd ee Sy ming a ad aio SarrSv 5 45 ro Sl. 000729"  L Slfeeekn: omrraeonenmadGagerasgrsamwoinen | L3b! 3S 3E31 ED eywaiw+ageo(esme2oan-ens === SFialmop:loeanUcIos 8 6379-135C: ETCAOD-NDAFSEOF-Uo1TNh2oD0MSaD)D.atatShuest5e99n1 86/04/4995 10:56 gs lp: ome $5 FPogGee .ooBfoe LL5 BaRan) furs: gE&eogl=Hl os% R vfgg % fs[9 E 1s2e PS i EF E Yao 5 em o o Yl 1194 mua = 1785 100. He wot] - 28 om spe a, A 20 ETAT 1972, Crt 38 5 ii e 6 s FY DEFIe NED 3a @ 0v g || =E=I | 1 {i 8| i{ v1 " R 83 FiSampBlrieeBe:sl0s4:9o69:5#6H320U-12315s; COLMPAS8Oa UBND8F-OT-Bh1ge7A8Mz SCSDABa)Eta[o8Shyose5t9Ee9m1LEY 86/84/1995 12:52 gpisse_gnel : Ss Sod ar 3 Ed op Fa i Fru an zoho [oH hers g $ oLka oTg onfF aa Tr ea28n f%oEFm fo&p F0i%obeaa o ape i i 200 [roe saz HP oe fers. lu 56 R 335 3 5 : F168 NOT DEFINED woos Fig oai ui vosstiinz/1Gs7T-E oi ween ou BROTGOPAVAILABLE a eSarmaptleorDLCDLLEIV RSutnraidPgarhtieent0Lsi5n-e0on8-Fi10t959-50f3o-r11c19e:3d359:t3Ah8TroVu1eg1rh:s3i)Oo:rn5i:3gin.& HE A E478 - Component 4:1 . py St /' + | / / WE: - / - wus LoL. ya ET 412215 4 8 [i EXTERNCAoLmpoSnTeAnNtDARD1 CA=LIBRATION EVEL AMOUNT AREA 21 55..81000000 377151853s 3i 11..26000000 16803737135 5s 54..00000000 7 12.0000 1ta6s6738085 535003 - - SIoPE aX + INTERCEPT AmouAnrte.a + +1.3353.7061-5050E-x 0x4 mAreeaant ++ 00..500D000UEE..0000 R squared = 0.9467 2 - 0007327" 9.11.3 Summary and raw data; ppm F" in serum as determined by thermal extraction followed by analysis using Orion ion analyzer. This data, although supportive, in the opinionofthe Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. 000733 " HWI 6329-135 AMDT 011095.1 DAnoahlrymsiasnnDaSteesr:u0m8/A0n2a/l9y5si-s08/18/95 ADlXl2s0e0r0umOrsgaamnpilcesHawleirdee tAhnearlmyazlelry aenxdtrcaocltleedcbteydainmoad1i:f1iemidllDiohQrwmaatnenr and TexIpSaAnBdabsolleutiioonn.anTalhyezesra.mplTehse wDeorhermmeaansnurweadsocnalainbrOatreidonusEiAng94304ppm, `40ppm, 62pm, for serum curve 11.00Tphpem,sa1m2e4pFpCm-,9525s0tpapnmda,rdasndweSr0e0puspemd FtCo-c9al5ibsrtaatnedatrhdes DThoehrOmrainonn wwiatshctahleiberxacteepdtbiyondiorfec2t5m0epapsmuraenmde5nt00wpipthmnfoorbslaenrkumcocrurrevceti2o.n uslsoipneg, 0i.n0te5rpcpempt,, 0a.n1dppcmor,re0l.a5tpiponm,we1r.e0prpemcoarndded1i.n5pthpemFappsrtoapnrdiaartdes.logTbhooek. A2).sAunmmainriytiatlabclaeliibsraitnicolnudceudr,vsehwoiwtihnsgttahnedaprpd mdeFviaitnioena,ch%RsaSmDp,leR(svcaelpuaege and equation of the line is on pages 8 - 11. Pwaegreesb3ei-n7g sanhaolwyztehde.exTcheel sDporheradmsahneentFtCha-t95waisnigtieanlecraaltiebdrawthioennctuhrevesawmapsles 0n.o0t5usisedbetloogwentehreaOteritohne data in the calibration asnprdeasdhsohueledt.be Any Orion considered reading below an estimate. TBhoveiFneC-s9e5ruinmitwiaalscuasliebdraatsibonlacnukrsvaen1d was QC spiked into bovine serum. check samples on 08/02/95 and 0t8h/a0n3t/h9e5.samHpolwese,vesre,rudmuecutrovper2obwlaesmsanwailtyhzbeldaunskisnhgarvaibnbgithisgehreurmrferadoimngs study serum # 6329-123 from study r#a6b3b2i#9t-1F25323r4a6b.bitOs nF5d2a3y4s60,8F/5125/39355,- 08/18/95 rabbit and F52332, were used for blanks and QC check samples sWteudwyewraesudniasbcloenttionluoecdataefdtesr edrauym8sbaemcpaluessefnoortDhainyg2wathsrfoouughndDianyt4h.e This first day of sampling. Page 1 of 11 000734 " STUD#Y 6329-135 SERUM FC120A8 HW 6328-135 Fluoride concentration in rabbit serum (ppm F-) GDrosoaugpe1: Omghkg FSEamZple _0D5ay31 Fs2o73 0408 0Da3y1s 0178 D0a5y8i3s D0a58y2z 0889 0712 FFssz2oo779s 00555117 0044330 00661724 0058214 FFs2200873 00457096 00322836 0088003 00467727 GDorsoaugpe2: 2mglg TSSamNple D1a4y6i Fao 06s 0D3a3y%s 0277 D0ay3s D0a6y1202 0483 0468 FFsszzee72 00444646 00320853 002038 0045328 FFss2a0f40 0043d8 0162977 003564 00667270 Group3 Sample _Day1 Days Days Dey2 Dosage: 200mg/kg FEF5s2sz9o8sm42 00.55904 058 00562954 0845 000222875117 00.662%0 0712 FFsezseto Fs 00s60422 0642 115170 0sf2 00624714 0777 00888M 0580 GDorsoaugpe: 1000mghg FSS2a6m7p8le D0.a7y612 Fas 101 0D4a4y9s 056 D0a77y7s D0a4yaz 0807 0S18 FFss22eoe71 Fs2088 018078 0549 00760%9 0478 00775020 0668 0083831 0249 Fs2005 0491 0513 0ST 0275 000735 ~--- -- STUDY # 6329-135 SERUM Sample rAecatduisnlg SaQmtpyle fTinIeSlAvBol msLpFiCkeOdS Cosnoclu.tFioCn95 recov%ery pAcptmuaFl- sMpaisksed recMoavsesred iD (ppm F) (mL org) (mL) BANK 00813On Zo {ppm _(uglug) in sample (ug Flug Fo mo BBLLAANNKK cooosme 0011 2200 00.779626 0o..00776882 BBLLAANNKK oooosttez 0o11 2200 00.862348 00..00862348 626.2PPPPMM..12 0000738354 0011 2200 000044 662 110288%% 115877 00114490 00.118577 25205.0P.PPMP-M12 0032094 0011 2200 0000044 225500 19001%% 650388 00660000 00650388 BLBLAANNKK 0c0o3z 0011 2200 11.8015 00..118018 F5B2L0A86NDKAY1 o00o7m2s3 o0i1 2200 01..64552 00..0164552 FF252000070DDAAYY11 0000232225 0011 2200 0.4645a0 00..00464540. FFs522008024DDAAYY11 0000220233 0011 2200 00..448466 o0l.00a4a6s6 FS6M2-IP0P0MA-1YI 000017904 0011 2200 004 6 102% 01.53828 0149 00..0135828 S2E5R0U-MPPBML-A1NK 00220476 0011 2200 00M 250 82% 44932 0600 0o4s91a2 SSEERRUUMMBBLLAANNKK23 00117067 00.11 2200 32.5125 00..231552 SSEERRUUMMBBLLAANNKK4S 000078425 0011 2200 11.6489 00..114698 SSEERRUUMMBBLLAANNKKS? 0000788%5 0011 2200 117511 o0/1n81 SSEERRUUMMBBLLAANNKKSS 0000669115 00..11 2200 113283 0o.z13s8 SSEERRUUMMBBLLANAKN-K110 0000568527 00..11 22.00 113118 0o13s1 SSEERRUUMMBBLLAANNKK1A32 0000677023 00.11 2200 114314 0o11a31s SSEERRUUMMBBLALNAKN-K1S4 0000668754 0011 2200 113315 00.113381 SSEERRUUMMBBLLAANNKK11S7 00000473 0011 2200 01.80618 00..0180618 SSEERRUUMMBBLLAANNKK11SS 0000430938 00..11 2200 0.0.870966 00..00870986 SER2UpMPBIL-A1NK20 000037s5 00.11 2200 0004 6 130% 01.79541 0149 00..0178841 e22ppPPuI23 0se0s2s 0011 2200 00000044 6622 11271%% 117840 00114499 00.117880 225500PPPPMM--21 00227602 0011 2200 00000044 225500 8907%% 552400 00660000 00.552480 FS208DAYI 00207 01 20 0.594 0.0594 000736". => STUD#Y 6329-135 SERUM Sample tAocntuianlg SeGmpyle {opm )_(mLorg) fTinIsSlAvSol (mb) msLpFiCkeOdS Cosnocl.utFioCnS oom) reco%very _tugiug) pApcmweFlin sample (sMvpagiskFse)d_ relcMuoagvseFsr)ed F-- swocoA-- vi 002s Fsmoavi ood 0011 2Te0 20 oosg6o8 o0.0w508 00.664022 00.00004022 FFoowoeosoaAvvii Foray ooogti ogi 0011 01 2200 20 00766422 0000786422 1.01 0.101 FFoosomiDDAAYY! FoSIwUeToOAR1vi 000050s7 0011 2200 Goeowe3 o0n1 2200 000 120% 018.807 178 0149 00..0180875 0.78 SPM mt on 20 0004 250 88% 528 0600 0528 Baannsieks e Banksewm2 000400%15 0011 2200 Banksewms ogre 01 20 00.783013 00.00783013 03558 0.058 FFooereedasytt Fszamidmi o0c0ds 0011 oo 01 2200 20 00.3459419 00..00459419 00.59383 00..0045938 FFoowrrooddeeyti OOoozzee Fogrsdet 002s 0011 01 2200 20 003555117 00.00551517 0479 00479 FFoeotsodadyi ee! 00o02280 0011 2200 Gos oi 20 004 4 1% 0057066 00% 00.00757058 sS1un0m0mbpopiima-nn1kk Sumbiank 000.0002908212 000.111 222.000 007 01 20 0.004 100 4 82% 001.3.4980633 0.240 000...10043908633 So% 00826934 0098 0000826934 TPeU, hel GGoois Goss 0o1i oi 2200 20 00004 4 Oo 0 ooss oi 20 004 To 791%% 0188s0 00204m0 00106%0 75% Ler 020 ool Seheell: For2DAYS G0o0e8ss 0o1i 2z0o 004 fo Fors DAYS OooBs4 01 20 ex% o1a8n7 020 000139171 0173 000.107430 TFsserrsoopaAvYss FozrepAYs ogooofgsst 0011 01 2200 20 00.445306 032 0000433283 0286 0.0285 FFosommoDpAAvvss FsoepAYS oooowt3 009 0011 01 2200 20 0013 01 20 - 00323787 - 0305 00.0023737 0.030 FFooooz7p0AAvYeS Skeezs 0o0ou2a 0011 2200 KE Stes oi zo o0Gs E@ s1ns% o02e8s3 171 oolmees 00.00628853 01% 211 oz oa shweszz OOS G1 20 ooos 2e 7i% 000737 ~--=, = STUD#Y 6226-135 SERUM Sample rAocatduianlg SaGmpyle fTiInSlvAoBl msLpFikCeSdS Cosnocl.itFoCnS reco%very poAcmwsPl- Msapisksed reMcoawsesed S REiDET oomF)_(mLorg) (md T (BopEm) S _(vgioq) insae mple (wg FIa lug ro FoBloandk s o00z63 0011 2200 T1.a2s7 00.112474 FFsofs0pdedanyes 0000M78 0011 2200 00..68374 00..00869947 Fs2daye 003 01 20 0526 0.0526 FFF5oo2s9er9o6addeyayyss8 o00.0o04s402072 000.111 222000 o0a1.t.814s05 000...0818114045 FFsosodadyss 0000762 0011 2200 0065142s o0..00851424 Faopondis 0000820 0011 2200 0004 40 110% o1s0s88 0096 00.10608 12B4lpapnmk-1 os 0.124 00.11 22.00 0.004 124 84% 21.4292 0.208 00..22429 FFF5o2e9s9e1sddeaysy8 000.003301554 000.111 222.000 FFeosdseeyts 000s21t7 | 0011 2200 FFss2T0a7d9daayyi1ss 00004308 0011 2200 FFoasrrseddaayyttss 00s0 0011 2200 Fswmpapdmayiis oOsodt2 0011 2200 steaupmpbimndk 000122838 0011 2200 sseknmebdioaink 00001938 0011 2200 sSpkkee4d0032 0000M03 0011 2200 ssppkheeti22442 0011232 0011 2200 ssepukmeb1l2a4n3k 0O0M02 0011 2200 FFss2238806 DDAAVViISS 000021861 OO0011 2200 FF52o8s07 DDAAYYISS 0O0o1M890 O0011 2200 FFsouoido DDAAVYISS 00007737 0011 2200 0000044 14204 00000044 4400 00000044 14204 00000044 112244 000.47670%89 000..000478703890 00551533 00..00585133 ofsrtz 00.00881529 00.6570s1 00..00580714 94% 00.850033 009 00.00350033 83% oa2t47s 0208 o0o.s2817a 78% 00.724058 0098 000.704358 8931%% 0078861 00.009985 0000788916 9802%% 224658 00229988 00..224658 78% 02.36035 0298 00..0283035 00349683 00..00348633 00.239680 00.00230680 -- oossse 00..00334554 FFF5oo29se8:s4 DDDAAAYVV1II5SS 000.0001121395 000.111 222.000 Fas DAYIS 007 01 20 000..222578711 000...000222705117 0274 0274 000738 ~---_ = STUDY # 6329-135 SERUM Sample. iD SPIKE 40-1 SPIKE 124-1 serum blank F52080dayls F52093day15 F52078day15 F52080day 15 F52091day15 F52087day15 F52088day15 F52005day15 F52072day22 F52073day22 4oppmspike-1 4oppmspike2 124ppmspiket F52079day22 F52075day22 F52076day22 F52983day22 F52986day22 F52080day22 F52097day22 F52082day22 FS2994day22 F53410day22 40ppmspke-1 16224ppppmmssppkiek-e1-1 serum blank F52084day22 FS2002day22 FS2906day22 F52077day22 F52080day22 F52003day22 40ppmspke-1 124ppmspke-1 serum blank serum blank `spike 40-1 spike 40-2 spike 40-3 spike 404 spike 124-1 spike 124-2 Actual Sample reading Qty (ppm F)_(mLorg) 00373 01 0152 01 00465 01 00785 01 ~~ 00389 O01 ~~ 00388 01 00404 0.1 ~~ 00376 0.1 ~~ 00350 0.1 00334 0.1 00285 01 00201 01 003% 01 00722 01 0.0460 01 0110 0.1 00280 01 00262 0.1 00236 0.1 00339 01 00305 01 00233 01 00264 O1 00219 O01 00310 01 00339 01 00645 0.1 0.430 01 00898 0.1 00405 01 00310 0.1 00318 01 0036 01 00343 01 00321 01 00200 01 00616 0.1 0416 01 0021 01 00172 01 00235 01 00333 01 00343 01 00371 01 oos2 01 00810 01 TISAB final vol (mL) 20 20 20 20 20 20 2.0 20 2.0 20 2.0 20 20 20 2.0 20 20 20 20 20 20 20 20 20 20 20 2.0 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 mLFC95 spiked 0004 0004 0004 0004 0004 0004 0004 0004 0004 0004 0004 0004 0004 0004 0004 0004 Conc. FC95 % solution recovery lppm)_(wolug) 40 76% 124 102% 40 150% 40 96% 124 74% 40 134% 124 88% 62 121% 40 128% 124 78% 40 49% 40 89% 40 71% 40 77% 124 38% 124 61% Actual ppm Fin sample 0.746 3.04 0.931 1.57 0.777 0.777 0.807 0.752 0.700 0.668 0.570 0.582 0.712 1.44 0919 2.20 0.561 0524 0472 0.677 0.610 0.466 0.529 0.438 0.620 0.677 1.29 261 1.80 0.810 0.620 0.636 0.712 0.685 0.641 0.580 1.23 232 0.442 0.344 0.471 0666 0686 0.743 1.12 182 Mass spiked (ug F) 0.096 0208 00009366 0.298 0096 0208 0.149 0096 0208 0096 0.096 0.096 0.096 0298 0208 Mass recovered (ug F.) 0.0746 0.304 0.0931 0.157 0.0777 0.0777 0.0807 0.0752 0.0700 0.0668 0.0570 0.0582 0.0712 00..0194149 0220 0.0561 0.0524 0.0472 0.0677 0.0610 0.0466 0.0529 0.0438 0.0620 0.0677 0.129 0.261 0.180 0.0810 0.0620 0.0636 0.0712 0.0685 0.0641 0.0580 0.123 0232 0.0442 0.0344 00471 0.0666 0.0686 0.0743 0.112 0.182 000739 === SamipDlo spkefT2R4S4 SSPPIIKKEE1000012 SPBILKAEIN0K03 FoB7L8A0NAKYZ2 FF5522008001DDAAYY2222 FFS02B0878DDAAYY22Z22 FsS2P0IK5ED4A0Y1Z2 SPKE100-1 STUD#Y 6329-135 SERUM rAocatduianlg SaQmtpylefinTaIlSAvBol msLpFikCeSdS Cosnolcu.tFioCnS5 recov%ery ppAcmwuFa-l sMpaisksed (ppm F)_(mLorg) (mt) OIE 01 20 (opm) _tuglug) insample (ug F) 00% TH 13% 410 CZs 000067899 0011 2200 00000044 112040 5564%% 113620 00229480 0O008B8St5 00.11 2200 00000044 110000 7711%% 117710 00224400 ooo4zss 0011 2200 00..454048 0000222558 0011 2200 00.541469 0000210608 00.11 2200 00.83315 000012358 00..11 2200 00..227459 000083530 0O011 2200 00000044 14000 7701%% 0167703 00204805 recMoavsesred lug 9 o04.T1o6--0 00117312 00.1097008 00..00884494 00..00558118 00..00323459 0000827735 6170 000750 "== + Nor SERUMCURVE SoSreenrAectmty wSoeeyree fT"aOiNvmSe mnLrecdesComno.tmtncss o_owpany tmAnun_sigewtots r"ybor nsg bmme ensdi ee OoomOoeSmr 000111 2000 gGdooowMw o 3H4 _eJogTmixeekgx) InL1lsieh0mmeele oocugoooFonm) ewarrimFnDmiseceotmeer Goom%M sSSammmis: Cwm GGGaoomwwe 000111 Goes 01 222000 dadooomm 20 dom 0dioo do etTouxxx 1Llio7wessw oocooosomeaoarrriaeasssseooenoceo ex ihm oem Gua 0o02nb1e essmmmtes GGooouoWs D0D111 322000 GaGomow @@@ ls1e0ok% vlLeeeeess ooonvveeers ocssrrsee:ewsmwoeeewnnor 00o05wm6e SGoemme:s ooOmMmeB 000111 231000 dGGooomM 01000 msxxx e1de0ms0 0oowmne0es2s ooamnaoekss simeemweeems 0o0mfml m2w69m9smz1 OOOIWuF1D011 212000 o0Go0omMw mm1se4 t95o2k%x SS27e4s2v0 0oo2a0a7n8 oe0.am2se7e4rs2 iSmToeEvveGsmex 0o02tv5s1a { s ooowwmeeesr oOoMmmTy 0DD111 212000 ooGooommw m 3mm0 oimoxw esereuiwes oGoeseooss coceesrsess smuesmmese ooosms ==I GoGpmmms ooodseass 0D0111 221000 ooooommm s=fmo xTaox SSgrememe lt1ai2me0esm osoeommme dsmaeemvcee oaco5e <g ql 8i | seman vows zn a . i. ii :is eE Fm il min F-- apep o0ot TbSee 0E 0 nmR erre xmxy aen Ee SmcaEEn EmSRmRoE pBoEm ds wn xI 4 mBi oRm EoomEoSweE im om sm on ee awn en cSMEmmR owEwmnEoon denBoEtmn a8OmZmo tW e osewHoemmsammeen e oewp crn mm ot 13 sm mn ese as seven see GEEEELGmEGN BBEE Bog OmRoEm BEim EERE SPKETMPPWT 019 01 EE 20 004 ID 1 100% o20r7 EmERr 0298 0297 somos cy0016 <5g5 ql | . For SERUMCURVE 2 NORMAN (un) STUDY exs-35 serum om _ gg im wo wo wn a a wo 3 vi 5 Fi 9.11.4 Summary and raw data; ppm F" in serum as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. This data, although supportive, in the opinion of the Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. 000 745-- ow lus Stato Qa RE: 6329-135 SERUM SAMPLES ADaMtDeoTf2A0n7a9l5y.s1is: 8/16, 8/21, and 8/22/95 Analyst: DDW `TThhee sgaamspilsecsolalreectbeud mined2.0inmtLheofDo1h:1rmTaISnABat/M9i5l0liC-Quwsaitnegr.0.T10hemLsoamfptlheessaerreumt,hen analyzed on a Skalar Segmented Flow Analyzer using the lon Specific Electrode (ISE) Method. TISAB buffer is added to each sample as it proceeds through the system. The sioanmpsleleectthievne geoleecstrtohdreouagnhdathheearteefdermeinxciengelceocitlrboedfeoirsemtehaespuorteednt.iaTlhbeetswigeneanl tihse amplified and related to the fluoride concentration. `pTphme ifnlusotrriudmee.ntThweasstcaanldibarradtecdurivnethfeorrtahnegehisgohfr0a.n0g1e5w-a0s.1p5loptptemd aunsidn0g.t1h5e- 1.50 inverse logarithm option. The standard curve for the low range is linear. All standards and samples were then calculated by the Skalar software using these curves. All results below 0.0001 ppm appear on the raw data as # #44. Aquality control standard was analyzed every 10 samples to check for accuracy and drift. aRnadwsduamtmaairsitzaekdenonfraonm tEhxecealppsrporperaidastheeecta.libTrhateedfinraalnrgeesuolftsthaereSkaadljaurstpreidntfoourtthe collection volume and any subsequent dilutions. 006 746~ b o se 31 COPY AVA on ILABLE SUSMEMRAURMYSOAFMP63L2E9S-135 AMDT 020795.1 LA IRE 4S ABT 2079S.) HwI 6329-135 S Kaloo DATA DoGseROLeUvePl1:0 Dose GLerveolu:e22mig Dose LeGvrelo:v2e0s0 mg GROUP4 Dose Level : 1000 mg/kg EEoomm 0L%451ND 013s% 0000ss EE2s9 0lse o0m 11009 oono;t EF20o8 00s % 004%6 009m1 0000s EEoos 011 o0s6 0023s 0000s EEorm o0s4 o0l% 0o2l3 0000s3 EEMoO 00ss 1186s 0023 oosm FEonss solm 0lo%s oomw 0000ss EEosm o0m l1es 00128 0000ss FE99s5 oomm 01ss 2L0i0z ooook EEtmo o12m5 0om% LLnm 0o0ms Easel 131 12 106 003 FF5o2s98s7 1Lo0z3 010s9 00.9954 o00m2 Fass 0 oss om 00 12555UMXLS 000747 FRET opA rbe i L camaro oan BE EE msmssersnsmonn t VHDiE oammtm geoem Swoeis oomnmn PS I oE EmEENo L2 2 JESTCOPY AVAILABLE BBBBBoooooowogsemmmmseEE an oogemmm oEksw wwmom iTo#T5 1% 18K62:1 38013 IPZ 20 010 Z EEEBRE2E2 Z 259 0004 00 ols 026 1% 1 g 2x i i B| om CmE mm5 F52082-1 002 seasee 0o0n 20 010 049 00ro oowo oows UEiWS:EE aEiiz8E sans 0N 2 on wseenn aaacnne 0B n0 oo oonn saimmn oomm maews odse aow ooww B M30 B Beuwwcsaeersnumm aoooms BBWooooonn aoimmw B$ o s Matna 1m aaLne om BooGkh Itnm eomm aanw ooms oonm uommer 45 sewaaamsn aoWonn I20 oolw sammo oomw aswe ooes oen wea bGPoor miamse: oooooxtt BBBo oaowomk soommm om deo ew on sen EoGobo FEraemmeoer oooooo Doaoonawn oaemnm bboo Fraaonem um aoaeosr m L0 oaon aemm a wan Io 5ba82 r1eSf2w0ee8e0n-n1 aaa0o0no8 2BBn0 ooaoke0o0 M1ii20nm5 aw aw os on wm o 9fo a G-n otwnmw om FR A 8g 2& BESTCOPYAVAILABLE IR Page sain Gupasssoan sans Dohm, ioeEsoSanAa7tX35TATWIDATASERMSSEAS 33Vo me a CPPooommmae AB ooSmaasr BFH ootes F 1P bb6oo BSPa RTeUhMeBkmL1K aioe iiwo woem Sowee oW be ane 0A m% mom ow om 2 o5m ow o0wo0ww8 os 2FBF0RR o01n0 a15 FR BESTCOPY AVAILABLE g FnRx gS %oEwE fl oo ' mwa on oot noi om 20 oo om . 1 DWiath 1% oLons lo won ox u$ag33 we oor 0 aw 1% oo wm oo on ww CHIE Paget Piet sans PB BEo Tma E 5NRe ) ome F Fior]T]om a5a ieixs " B8o rmmsass i8bo maannsan oa2D hEo"wshes Im 2SS Fh1w er Faaso F iH r nsni o g o4FR mmaas g>3o6@GeoDmrmifat ` ooann ooomnn ue aoRnoem oo>nn obomr ooonne we oo0nn% wm io o0oo0iin3 ooorou ooonums 100 o1Wwns om 0BBoooown B3o0wmok n3 oe0h 1nF0R oaohm nBBooomhn FFBRR oob no ooamna em we oiIbmn aowe elmm Hdk oomommeme aeme aw wes toow mn ooke om a um ooombw ooohnh nbee ow om om um B32Bo0 0o0oo0kll0 0oat6mnh1 oamm aamw oosn oinh iwek IBno ooolool adamnw ooime emeonow oinp ibekr Bnooooonoh idimwm 2n0 oanoe Page BESTCOPY AVJAILABLE sas o a baasnmss ooouwe 0 w oooono eo1m% 65a hESaamss ooomwm no sean an 0Bo0w0oon ooinem ow aw ow es sm ol 3% om me am en hm aHx abiak uo on n m Lom im nnXooo remossws oooomw wooooonn ooimmm 5B[ o i rrea ooomoort BBBooooownn ooimwe 55Wo mmaeemens ooamuot Bnooooonno odimwm 8uv mVeTaaan ue oiwnns ow no om %B5 emnea e n oon no w 200 o0l0e 3i%n oowe emwe aoow aans meew a ah w 8g 3 So Ho BESTCOPY AVAILABLE Page3. Ospeor spow oupautsousn nssoxs DuAcrtcA ane:v CHpOKsAi nLA1R2D2A9TAUNTDATASERUMOS3131 :3Lom[Tea aiwn oiioe am Somsmammnn Po Sema eW omes Saooen am on om doemw 5so o a SSsm ooonahoooohow ouowkwe BESTCOPY AVAILABLE BBWooo omSsewnemn eidlbo oiiemn ooowwk ui. iT=rar in oFwnoy oe 0 ow os xF [ bo ry i eSasr s oaaosnno 3 am a I320o0a0Gon0hw oooImbmn oeommm wwwmem ooibnh oaimen wwamme Boal ax om me ok ok oer g nFoR os ry ooain EE on Bowoohh odanmm oomm mae oonn aann noon vr $x TER oe lo 233d EE "a n Wash FS2990-15 ND o00n2 2W0 oo0n10 o03n3 "So3end: Il Paget 53t nssoxs n03o Fraomamaess aaaonnn 0 awawnn ooonnn 5N3o rpmoemens ooanno Xo mam on BBBooaoannb ooonnn ob om 3aBo wVaa aen m oGwm8 se 0 00 om ew ww ow ow ses @B 8 n mmask ooowno bo rma oo 0Boooononn aaamme Boon im Sio rcraees bo ran oaoonees om BBooooonloo oiinwm Bo ol om a mo BESTCOPY AVAILABLE bPo avna aw aodooest oem nBo omob oomn 535 mamfnayn powioodn S00 en oo n020o00oa0n o0imn em wm ow om am S50o sSaoien oooiwn 5 mn ont n0 ooooooll IIimn dcee lmme aonw oohb bsee nan om o oG5 rraemmsnm iiIroor F 0BanonTooas ggS a4a a[mo ien e otWowar aw 0 ww om meen ow FR I Prez ssoxs wGGrraaennna ao0or oor 0wowooww ooommw onnoo raSowwmaen no aa o00ns aann L30 owaabb o1mm 3% oomm wmma oonw oomn dwoewr 3 ob 2a om am os on dow n 5 bVxaa am alwooo ww Loe im Nnnooo orrwaannn aaoo00sss 0Bo ooonkn ooommw nno amrreaann oo0o0os Lanon on ooommm Bn"oo ssoabmrabe oaens m B00aw aAmm oeowm emwe oolw oonm oowe 8 Wa wo 4832 * Pes mvaenrson powont -- Dmo i N sat s20640WA1 Sie Sm A re ED TO ee rate !Doom m au2 --z ?3 wSheHm w uapm oO mw www 3I EEEEonOm oosmo om }:b5 oSE E mmadsf Lowis owoo2n oBmobme BEST COPY AVAILABLE PPy o oe=m mBoR m5 E i oo 0 ow om P:1:" SEU=IsMaB=nLmK2 0o2-0n3 2BBB0 oootikossl 0oo&tp7e outwmw omamxm ootaom mooaw omuowm 2[[ I ge Fo=r] O BP ooEkR EamEuomm omanm 4 oono ono oo Fog =u =u E BoS w EomoD dmoe m oe omsow TE geo oLr3Fomm oIRuY ow 0=5B w0 owaw oam omaw woem eoxm eonm owxm iP5TSaagg gor wm = Bomomodm om om - oom 52pdi 1ssoxs e a3 axx aanos 0 aoak aosm J N%o Emweean oaaon0r wBoaobaonmoonooos %55 bpamDwannn ue o01ms ox I oawe oonm oGo oweaa[mnn 00wo 0 ow as a" aio[weY um osn um 0 om am em wo om ea ew g == 2g 2 4 Ih| Paez SERU7M-31C.U9R5VE 1 NORMAN QMOOT 2re0s7H8S5 WoT 325-13 SolanQuan Supe RSelulrt Bm DIfTouIlSvoAlD mSLoFhCnS FCC9o5neSn GD Ske Gp) SMpakuesd 8F) AMwaosgse Reco%very Reo oF) Sskla2 SA 00001 9.2200 oo 20 0o0o4 oo 30000 30 oo ois lk SSkkiot2 0o0m1 2200 0o0omd 4a000 00 ois Is Ses om 20 om doo sSkwaa1z Skas o0m om 2200 20 oooomm aemm oom am ois ols mm sSkMi0e2r oomi Skies om 2200 20 0o0m4 1000000 oo 100 0 038 SSkwieir oolms Skies om 2200 20 o0o0d m1400 oom 10 0m 03 us Ssmaolz o02m% Skzws om 2200 20 oooo 3250000 oo 2:00 0m oa lm Ssasml: Shaws 00474.2200 osm 20 oooo 0 300 oo 300 10 0 mw SERUM CURVJEa1m(NeOoRnMsAN) 731.95 eon i i H C EHr1 i TTT] PE [rer SERCRVINAVE 600758 `SERUM CURVE 1 7-31-95 NORMAN TmI qe cAmoE n WroETnoSme,nIdaRn lWdEw L EmEu om oom oumom on mm --osm_ ue fmSkeearomm00o8ommo20oomm00o0o4 aa4mn000 o 0o1o0n ofOeI6 owIne% me g mne STANDARDDEVIATION: gm00826 wEdaTarTo om o3n osmmoeam ooB mn oowu obmm st wowoor sm oSgooen mawnnonnowommme ommoeomwoomEw omoomn owwem eowmoeogmsoon gouns ggEiosnosooewanmmoemowoompo omw ooaNnw Wuummm soeeespwmeon n am Bmeam mwoonn oemn ownn on ommmomm oomwm 3o0 wwme ommeomum eoxm ognR ogee gn nmeomesoempnese or ooavn Hao ox omom 32 oR Sr iw-- n s rrr er Poe | = II rrr 'SERCRVIN.SUM 600759 = -- ncoen war on 20 ou er on 20 om om 3EST COPY AVAILABLE W meEiBoEm z . Smen ti r TT T T T[A fol] at bl goer LZ Jol 44 Sal | T| fou TT Rew 1TT| wxcnnne y | ' bi2) gE coeregTM SERU5M16C9U5RVE 2 NORMAN pe"1 COPY AVAILABLE TswEcNssDi Moaoe d0doe kSaLi el Gtomfe 332000 saasi el aedomos Z2do0o al WB hiSeoelerss ooonum 323000 SSelieesd Teles oooiul 333000 sSkiher TEED oo0inh 233000 ooomame muM0m0omOomm aaGinul eeDewF STANDArRDoDE!VIATIN: f1i7r oo0m6me4 d4seo0mm ooGowi ao0mm;s 0ie%mm STANDARrDsDpE!VIATN: oomsue ooommme o@amm aa0sns ooGlwmo m6o%mm STANDARrDsDpE!VIATON: w0o4mm om am as om wm ooaooom ww1e0o0 o0om3a a0omwm l1sWww% STANpDArRDoDEVITON: s0o2ss acGoooo ewwooo ooomaa ooommm bnewew o0G0omM 0mmoo o0om%m o0o3ms omwk SIANDARDnDsEoVIATION: f0liers]t mew SERUMCURVE 2 (NORM$A1N69)5 yma TT -- A sol [TT a YT TerrTT] wet EY sencrvansn 600761 Fat 1995-08-16 14:10 OutPut of : 950816A1 Software : version 6.1 c1990,93 operator + bow Date of the Analysis : 1995-08-16 08:58 Analysis File Name C:\SKALAR\DATA\HWIDATA\SERUM\950816A1 000 "halts, AudT 2073S:1 HA WE 6329-1T3o5k FClauloirbirdaetio1n.5order = Inverse Logarithm s10p0 is sarees [x=-=a] Cx1 == ccoorrrreecctteedd vvaalluuee ooff tthhee csaomnpcleentration 1 Result = 100s 5 = Slope of the electrode aai2== -00..0000006050 20 -1l15706 FCalluiobrriadteioLn order = 2 Correlation : r = 0.99717 Result =a2 * x! +al * x +a0 aai2== 00..0000000108 20 = 0.00912 sampler Nyupmeber + 15A1000 VSaasmhple TTiimnee i: 51020 sseecc.. MTarke upmime : S1inglseec. spacial : None needle Height : 70 mm. Diluter dnoioldulteionHeiFgahcttor :: 1800 mm Raielsuatmipolne Volume : 12.5 ml. Dilution runs : 1 uRseeprrocfeisle :: No . mr 600762 ~-- 1995-08-16 14:10 OutPut of : Fluoride 1.5 sPiagtnhalnumtybpeer :: De3bubbled DesycsotleomrNumber :: Y0es dRielsuatmeple :: NNoo 4d1i6l oTuhtrpeusthold:: 04095 Window event : Off 5s21 ssTTaannddaarrdd :: IIggnnoorree ss3i ssTTaannddaarrdd :: IIggnnoorree $s65 ssTTaannddaarrdd :: Igno0r.e150 ss78 ssTTaannddaarrdd :: 00..630000 5910 ssTTaannddaarrdd :: 11..250000 DOridmeernsi: onInv: erPsPeM Logarithm sttrairgtgerValLuiemit :: 5108000 DSUec PsotaAkrtsihgnaorpee:: P6o0inteSdec MoeNadsureigwnionrdeow :: 17205%Sec RFielgteenreration =:: NNoo ofourntuplaut:: #4.44% Fluoride L PSaitghnalnumtbyepre :: De0bubbled DesycsotleomrNumber:i N0o diReLsuatmeple i: NNoo diiGl Tohurtepusthold :: 04095 Window event : Off 950816A1 000763 ~-- 1995-08-16 14:10 OutPut of : ss21 ssTTaannddaarrdd :: 00..003105 s543 ssTTaannddaarrdd :: 00..009600 5s65 ssTTaannddaarrdd :: 00..115200 s578 ssTTaannddaarrdd :: IIggnnoorree 5$190 ssTTaannddaarrdd :: IIggnnoorree oDridmeernsi: on2 : PPM tstrairgtgerValLuiemit :: 1580000 DSUec PsetaAkrtsihagpneore :: 6P0ointeSdec MeeNadsureigwniornedow :: 71205%Sec RFielgteenreration :: NNoo ofourtnUplaut:: c44.:8=8c8d4 950816A1 000764 -- 1995-08-16 14:10 Output of : 950816A1 Page 1 of 2 Fluoride 1.5 Fluoride L PEM Pou Pos Typ Ident Ch Result Time Ch Result Time wt oitw InitialTraWcaesrh 33 01..047702 20675 4401..08039315 00 23 aow 3 51 DWriafsth 33 01..047806 standar1d 3 0.074 368139 4401..08059518 729 40.0164 00 0 56 ss23 SSttaannddaarr32dd 33 00..100801 1098017 4400..00260811 00 78 s854 SSttaannddaarr45dd 33 00..111377 11245384 44 00..01822914 00 130 s5s7 11 8 SSttaannddaarr67dd 33 00..125845 Standar8d 3 0.614 11670884 4400..31440808 1958 4 0.7483 00 0 1123 5s910 stSatnadnarddar1d90 33 11..241797 14 a Drift 3 1.490 22310383 441.1.48648472 2484 4 1.8617 00 0 516 ouw SPK 6W2a-s1h 33 00..017402 22782360 44 00..01029916 17 u SPK 250-1 3 0.346 3011 4 0.4211 00 0 1189 uu SPK B2L5A0-N2K 33 00..312326 33138671 44 00..13280962 20 u BLANK 3 0.105 3536 4 0.0686 00 0 2212 uu P529B8L6A-N1K 33 00..101878 33781806 44 00..00481809 00 2234 uu 25 u FF5522999907--11 33 00..007886 P52982-1 3 0.079 44026326 4400..00233747 4410 40.0247 00 0 2267 wa 28 u DWriafsth 33 10..047707 F52994-1 3 0.081 44852856 44 01..08049113 4934 4 0.0281 00 0 2390 uu PSe5k341602--11 33 00..017282 55121808 440.0.00292788 3 ou SPK 250-1 3 0.294 5462 4 0.3525 00 0 3323 uu BBLLAANNKK SSEERRUUMM 33 00..100915 55683150 440.0.00561295 00 . 3345 uu BBLLAANNKK SSEERRUUMM 33 00..009983 65196816 440.0.00458656 00 3367 uu SSPPKK 6622--12 33 00..114328 66531305 4400..11410405 00 3389 wa DWriafsth 33 10.047805 66961814 44 01..08059319 00 1a0 uu SPSKek25602--13 33 00..123564 77201304 4400..12199741 42 u SPK 250-2 3 0.271 7382 4 0.3208 000 443 oouu 45 u P5522998942-11 33 00..008885 52996-1 3 0.082 77375374 440.0.00430576 7910 40.0310 00 0 3475 uu FF5522999874--11 33 00..008856 88028568 4400..00335664- 00 i49 uu 50 a FF5522999738--11 33 00..008858 Drift 3 1.488 88463048 4400..00430478 8785 4 1.8587 00 0 5512 wu 53 u F529W80a-s1h 33 00..017001 F52991-1 3 0.103 99103014 44 00..00069215 9311 40.0654 00 - 600765 -- 1995-08-16 14:10 Output of : 950816A1 Fluoride 1.5 Pluoride L PEM EY Pos Typ Ident Ch Result F Time Ch Result ? Time 5545 uu s$p5k298672--11 33 00..102975 99646826 44 00..01501571 00 557 au PK D2r5i0f-t1 33 10.1520537 109081323 44 01..38081371 00 w5t8 wrw BunOut WWaasshh 33 00..007700 1100428477 4400..00009911 0o Page 2 of 2 006766 -- Calibration curve of S50616A1: Fluoride L 0.2892561 1| //|) || :Mi l | || ) 0.009110 0 order: 2 | eased riosn soo 600767 ~ |||lvosawrst | | | | | t || | BEST COPY AVAII ABLE Calibration curve of 95081601 Fluoride 1.5 -------------- J7A ya 7 || | | { | | "| 0.0898451 5 Order Inverse Logarithm[-- so sen 05 600768 - ~--- A055 || x fo cts of SIS Farid 15 on | w I|| ||| | |i 3] | | | | | | ] af oH 0 "A o Te S000 Esc=Exit | Fi=Help | Crtl-P=Edit peaks ! 000769 _ 4095 1 - Joe | BEST COPY AVAILABLE Rau data of 950816A1 : Fluoride 1.5 " @ | | $ i - IN Bo | kK ANS As | |e 04 as60 Tine EsceExit { FizHelp | Crtl-P=Edit peaks | vno 8 9560 660770 _ ws Pathno- 3 | A Tine: IEEESM Value: BESS DeltGaE:N o | | | | 3 i Pd | dy 4 WU a ' Cott T Fiat TTC pea T 606771 TT 1995-08-21 12:30 output of : 950821AL Software : version 6.1 c1990,93 operator + Dow Date of the Analysis : 1995-08-21 07:55 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950821A1 Cou as kv 20 hws 329 Sern lians 2 CFalluiobrridaetio1n.5order = Inverse Logarithm slope is = aren [x=-=e] x1 == ccoorrrreecctteedd vvaalluuee ooff tthhee csaomnpcleentration 1 Result = 1b 5 = Slope of the electrode aa2== -00..0000007010 20 = -1.20061 FClauloirbirdaetioLn order = 2 Correlation : r = 0.99953 Result = a2 * x? + al * x + a0 aazi== -00..0000000205 a0 = 0l00771 Sampler Nuympbeer i+ asAL000 WSaasmhple TTiimmee :: 15200 sseecc.. Mr Time 1 sec. spnTaeekecedilaeulpHeight ::: 7NS0oinnegmlae. Diluter 4neieldulteionHeiFgahcttor :: 1800 mm Rdeislaumtpiloen Volume P: i2.5 ml. Dilution runs : 1 URsoeprrocfeisle :: No . mT 0600772 ~-- 1995-08-21 12:30 Output of : Fluoride 1.5 PSaitghnalnumtbypeer :: De3bubbled DseycsotleomrNumber i: 0Yes dReisLaumtpele i: NNoo d4iilG Tohuretsphoultd:: 04095 Window event : Off ss21 ssTTaannddaarrdd :: IIggnnoorree ss43 ssTTaannddaarrdd :: IIggnnoorree ss56 ssTTaannddaarrdd :: Igno0r.e150 s578 sstTaannddaarrdd :: 00..360000 55910 ssTTaannddaarrdd :: 11..250000 ODridmeernsi: onInv: ePrPsMe Logarithm sttrairgtgervalLuiemit :: 5108000 DSUec SPteAarktsihgnaorpee:: P60ointeSdec MeeNaasureigwnionrdeow :: 17250 Sec FRielgteenreration :: NNoo ofourmtuplaut:: #H.4#H Fluoride L Psaitgnhalnumtbyepre :: De0bubbled DseycsotleonrNumber :: N0o aiReLsuatneple :: NNoo 4aiilG oTuhtrpeusthold :: 04095 Window event : Off 950821A1 000773 1995-08-21 12:30 output of ; a3515 SsSTtmaaannndddaaarrrada:| 000...000531005 3S5i 55 oSSrrteeannndddaaarrrdad:ii 000...01235000 23573 SSSrtteaannndddaaarrrddd :: : IIIgggnnnooorrreee olDriodneern'sS:iToan2pda:rdBPH: Ignore FSStieaiargteeoshrvaaplTeui.enie ii 1P300o00i0ntDeSUdec NSTeaaarseureligownnioornreedow i: 176705.0 s3Seecc oRFiemlgnteeinraerattieoans:ticvaNeo Cpu: ener 950821A1 600774 . 1995-08-21 12:30 output of : 950821A1 Fluoride 1.5 Fluoride L PPM Pry Pos Typ Ident Ch Result Time Ch Result P Time w1 t iw InitialTrWaacsehr 33 10..046638 20685 4400..50906727 00 32 oaw DWraisfht 33 10..046832 538720 4400..05097972 00 5i s5z1 Ssttaannddaarr12dd 33 00..007836 970238 440.0.00219484 00 7 s543 SSttaannddaarr43dd 33 00..100960 11205893 440.0.00682941 00 5 s8e5 SSttaannddaarr65dd 33 00..115268 11640395 440.0.11510894 00 1110 8s7 SSttaannddaarr87dd 33 00..621799 11975883 4400..32943850 00 1123 5s910 SStatnadnadrdar19d0 33 11..427214 1a Drift 3 1.539 22133039 440.0.55946388 2483 40.6121 00 0 156 uow SERUMBLWKash1 33 00..006938 22873223 4400..00707675 0 1178 uu SSEERRUUMM BBLLKK 32 33 00..008962 33011824 4400..00565729 0 2190 uu P5522998985-11 33 00..008813 33356304 4400..00451602 0 2212 nuu F$5522997723--11 33 00..008824 33781805 4400..00451888 00 2243 uu F5522997759--11 33 00..008803 44026304 440.0.00540480 00 225 au F529D7r6i-f1t 33 01..057299 44431803 4400..06402909 00 2278 uw 520W83a-s1h 33 00..006832 44891303 4400..00047871 00 3209 uu SPSKPK14000--11 33 00..104972 55121815 440.0.01647124 00 3312 ouu BBLLKK--12 33 00..007754 55466302 4400..00334272 00 333 uou SPKBL4K0--31 33 00..009710 55891808 4400..00824463 00 336 uou SPSKPK10400--12 33 00..103833 66313662 440.0.01621571 00 3378 aou SPK D1r0i0-f2t 33 01..153410 66658142 4400..16122231 00 1390 uw SPK 1W0a0-s3h 33 00..104623 76093072 4400..10307572 00 4a12 uou 43 ou $52972B-L8K 33 00..007715 F52973-8 3 0.064 77230874 440.0.00235467 7558 40.0090 000 4454 oouu F$5522997759--88 33 00..007775 77970396 440.0.00334878 00 4476 uu $5522998763-88 33 00..007701 88205875 4400..0022613 00 4498 ouu 5522999860-88 33 00..007701 88640397 440.0.00223711 00 5510 wa 52 u DWraisfth 33 01..056235 52997-8 3 0.073 98071825 4400..06007370 9136 4 0.0303 00 0 53 u F52982-8 3 0.070 9310 40.0248 0 Page 1 of 2 - 600775 1995-08-21 12:30 output of : 950821A1 Fluoride 1.5 Fluoride L 2 Pou Pos Typ Ident Ch Result ? Time 5554 uu 5u SSPPKK 6622--21 33 00..108254 99646848 SPK 124-1 3 0.144 9838 5578 uu SPK 124B-L2K 33 00..111761 1100108194 5509 uu 5523949140--83 33 00..018095 1100533695 6612 au P52D9r8i4f-t8 33 01..058542 1100878158 6634 uw $529W9a2-s8h 33 00..006832 1111213249 6665 uu $$5522999976--88 33 00..019012 1111548195 667 uu $$5522909839--88 33 00..008920 1111973686 760 uu $$5522998708--88 33 00..008780 1122219008 772 ouu SSPPKK12440--11 33 00..106968 1122644626 7743 au DriBfLtK 33 10..513066 1122989106 7765 ouw 5299W1a-s8h 33 00..008693 1133324312 777 uu 552290887--88 33 00..007895 1133658137 8709 uu $$5522997925--185 33 00..007890 1134806472 8812 uu $$5522997793--1155 33 00..009833 1144231982 8813 uu $$5522997756--1155 33 00..008758 1144576460 8865 ua $5298D3r-i1f5t 33 01..058704 1154091962 8878 wu SPK W4a0s-h1 33 00..006938 1155363441 8590 au SPK 1D2r4i-ft1 33 01..156354 1155671992 Wot wrw RunOut WWaasshh 33 00..006633 1166023627 Ch Result Time 440.0.01514395 00 4400..11367539 00 44 00..10093277 00 44 00..00552372 00 4400..60105737 00 4400..00644769 00 44 00..00862239 00 4400..00448180 00 4400..00475526 00 44 00..10680964 00 44 00..60101757 00 440.0.00651424 0o 4400..00442302 00 4400..00647492 00 4400..00554071 00 4400..00445140 00 4400..60200747 00 44 00..10670611 00 44 00..06017170 00 4 0.0077 0 Page 2 of 2 000776 Calibration curve of 9SOR21A Fluoride L 0 25200 | EE V4 | Channel 0 Measured: IEE Concentration: [MIIEKIEN Delta: CXR / 3 {t! 3 0.077061 0 order : 2 eased + 0.9585 wo 60077 --- [---------------- Calibration curve of 50621A1 Fluoride 1.5 1.7854700 Channel 3 Heasured: WN Concentration: MNIESER Delta: (Zh | 3 {1 t Qg 0.063020 0 Measured Order Inverse Logarithn 5: 586 035 000778 ~--= [ZITTHNENNTTE 0 WOT 756 || ot eirded) |i | ! | : |9 < W 0 3| Te 2% I| | | wo|o1 600779 ~--= aos . " A 1 i: | oes Aan |s(a14 e4 ] s| SoetAt nog jaanA | 4560 | Tine 9560 000750--.- |= [IEEET PY Ges TITHET Ga] E TT | || | | qo| i |EY | 1 | i.iJ l s RA AAAI Lette ali an Joe ` 8935 Tine 13935 sous Pathno: 3 Time: HEEIUR Value: BENZ Delta: [XN | | [a | | | 3 . | AAA PaaspP> agll A SA . 600782 1995-08-21 16:58 output of : 95082181 Sooftware : version 6.1 c1990,93 operator + bow Date of the Analysis : 1995-08-21 12:29 Analysis Pile Name C:\SKALAR\DATA\HWIDATA\SERUM\95082131 Qos Blashes, RAOmoTT03220m99-53.51 SeavmComme T- CFalluiorbirdaetio1n.5order = Inverse Logarithm s1ope pss sarin Result = 10b [x=-5=o] x1== 5= CSclooorrprreeeccttofeeddthvveaalluueeelecootffrottdhheee scaomnpcleentration 1 a3az0:== -0o.l0o0o000s0s -1.19997 : PClauloirbirdaetioLn order = 2 Correlation : r = 0.99960 Res=ua2l*tx? +al * x + a0 aa12:= -00..0000002050 20 000307 sampler Hyupneber + WSaasmhple TTiimmee :: MTsaprkeeciaulptime : needle Height : Diluter dndeiiellduultteiioonnHeiFVgaohclttuomre ReDsilauntpiloen runs URseeprrocfeisle :: No 185A0100s0ec. 1120 sseecc.! sXoinnegle 70 mm. :: 1800 mm i:: i21.5 ml. . mr 000783 - ~--- 1995-08-21 16:58 output of : Fluoride 1.5 DSPeiacgtonhlaolnrumtbyepre |: YDee3sbubbled RaSieytssuattmoepnleNumber ; i l0 o MNo Wd4i1indooThwuretesvphenoutlta::; 0o4f0f9s 1523 SSsTTtaaannndddaearrrddd :: ; IIIaggmnnooorrreee 2%555 SSStTraaannndddaaarrrddd :|i TIagmnoo0rr.ee150 S3o7 sSmtaannddaarrad:: 00..630000 O5Sr9ldoer'oS:rtaannTddnaavrreddrse:i Log11a..r32i00t00h SDttiramiregtngseirvoanlLuiem itFP :|: 1508000 DSUec nPSetaaakrc'siihgghnaoorrpeee: P2G0o.0inteSSedecc FiMolatseurre window Regeneration :|iNo7o5 4 Sfoprumrula:inane Fluoride L DPSaalctoonhtaoinrutmyepre E: DSe0bubblea alRSotyssuattmeepmleNumber :iioo0 Wd1ii1nsdooTwhuretesvpnenoutlta:|| o01f0f95 95082181 000784 __ 1995-08-21 16:58 Output of : 950821B1 . ss12 ssTTaannddaarrdd :: 00..001350 53 s4 ssTTaannddaarrdd :: 00..006900 s5 56 ssTTaannddaarrdd :: 00..112500 ss87 ssTTaannddaarrdd :: IIggnnoorree 55910 ssTTaannddaarrdd :: IIggnnoorree oDridmeernsi: on2 : PPM sttrairgtgerValLuiemit :: 1580000 DSUec PsotaAkrtshiagpneore :: P6o0inteSdec oMNedasureigwnoirnedow :: 17205%Sec FRielgteenreration :i NNoo ofourmtuplaut:: c#48:8=8c43 600785 .--- 1995-08-21 16:58 output of : 95082181 Fluoride 1.5 Pluoride L PEM PEM Pos Typ w1t ciw 23 oaw 35 s521 7 34 &9 3s5s J1o1 s57s 1132 3s510 1145 wa 1176 uu 1189 uu 2201 uwu 2223 uu 2245 uu 2276 wa 2298 uu 3301 uou 3323 wuu 335 uu 3367 uu 3389 wa 0auu 4432 uou i45 uu 4467 uu 448 uou 5510 oaw 5523 uu Ident Ch Result P Time InitialTraWcaesrh 33 01..046838 20685 DWraisfht 33 10..407683 632854 ssttaannddaarr12dd 33 00..007848 971313 SSttaanndadradr4d3 33 00..100991 11028615 SSttaannddaarr65dd 33 00..115260 11463151 ssttaannddaarr78dd 33 00..622709 11798670 stSatnadnarddar91d0 33 11..242742 22311315 DWriafsth 33 10..046335 22468652 SSEERRUUMM BBLLKK 21 33 00..007750 32081337 SSPPKK 4400--21 33 00..008810 33138662 PSKPK12440--13 33 00..104856 33573164 SSPPRK 112244--32 33 00..114550 43086828 5296-B1L5K 33 00..006776 44421318 DWriafsth 33 01..046038 44753826 $$5522999907--1155 33 00..006679 45190398 $$5522098924--1155 33 00..006649 55425895 $F5523948140--1155 33 00..006658 55861315 $F5522999962--1155 33 00..006656 65136837 5S2P9K74-01-51 33 00..006861 66531334 DWraisfth 33 10..046334 66698264 sPK 124B-L1K 33 00..019971 77023162 5p522909893-1155 33 00..101888 77358569 5522997808-1155 33 00..008889 77793161 5$522999871-1155 33 00..008836 88206833 P$5522098985--1155 33 00..008739 88463173 DWraisfht 33 01..405643 89072847 F5522997723--2222 33 00..008835 99311337 Ch Result F Time 4400..06033418 00 44 00..06033212 00 4400..00125874 00 44 00..00960132 00 4400..11510871 00 4400..24501673 00 4400..56730038 00 4400..60203113 00 440.0.00138144 00 44 00..00440361 00 44 00..01531851 00 44 00..11388461 00 44 00..00132276 00 44 00..60104361 00 44 00..00111654 00 4400..00015687 00 4400..00103770 00 4400..00009803 00 44 00..00413110 00 44 00..60201311 00 44 00..01781551 00 44 00..10053661 00 44 00..00558559 00 4400..0054306 00 44 00..00437845 00 44 00..60206331 00 44 00..00541517 00 Page 1 of 2 - - 000786~ 1995-08-21 16:58 output of : 950821B1 Fluoride 1.5 Fluoride L 12 PrM Pos Typ Ident Ch Result P Time Ch Result F Time 5545 uu SSPPKK 4400--12 33 00..019137. 99648692 44 00..01605157 00 5567 uu SPK 12B4L-K1 33 00..104960 190803180 44 00..01539931 58 u $52979-22 3 0.080 10186 40.0411 00 0 5690 uu $5322097756--2222 33 00..007776 1100356357 4400..00334214 61 u $52983-22 3 0.082 10713 40.0453 00 0 6623 wa DWraisfht 33 10..401693 1110182806 4400..60103715 64 u $52986-22 3 0.082 11237 40.0438 00 0 6656 uu $$5522999970--2222 33 00..007787 1111450893 4400..00334633 67 u $52982-22 3 0.074 11762 40.0284 00 0 668 uu $5523949140--2222 33 00..008823 1121191346 4400..00444650 70 u SPK 40-1 3 0.108 12288 40.0893 00 0 7712 ouu 73 ou SPsKpk12642--11 33 00..116381 1122466328 4400..11261217 BLK 3 0.091 12812 4 0.0621 00 0 7745 oaw DWraisfht 33 01..046639 1132292875 4400..60300311 76 u $52984-22 3 0.083 13337 40.0469 o0 0 7778 uu $5522999926--2222 33 00..008824 1133561815 4400..00445759 79 u $52077-22 3 0.083 13858 40.0467 00 0 8801 uu $$5522999839--2222 33 00..007892 1144023151 4400..00434907 82 u SPK 40-1 3 0.106 14379 40.0857 00 0 883s ua SPK D1r2i4-f1t 33 10..145621 1144733559 4400..16425890 85 w Wash 3 0.063 14976 40.0031 00 0 Wt rw RunOut Wash 3 0.063 15210 40.0031 Page 2 of 2 60678_7_ Calibration curve of 85062181 : Fluoride L | TTT NT0iE E MEN | ~ J 7 :: { 8 0.0030712 5 order 2 Wessured r: 0.0030 500 600788 ~-- | 1.7658142 | | 1 | 3:{ | 0.0831072 0 | 1 So /| / ! | / | | Measured 2085 C00789 ___. ws FCTwE aw dats of SSUGEIRY Flunride 1.5 meee 0 WOTH soc poirHedl RY wv | i 3 | :. "a " } 3 os | o Sa wd RAal ot ssi] | oy CecrEurt Fite GOITER peaks G00790 ~--= Rau data of 95082181 : Fluoride 1.5 ass # | 23i 1* oh Ap Anoa a EN PAS wag | |5 | SAve 0 1560 Tine 9560 Escobxit | FivHeln | Crtl-PeEdit peaks 600791 | sss ZTE fo ETT dotaof - = SORRY Flore 15 -------------- [PYITH 730 reed] | i:i | NE & | a8 hella gw| lod owasSadH I ARAN dgnAa 0 oss Tie os Eb 1 Fioheln | Gru peaks 000792--.- Raw data of 95082181 : Fluoride 1.5 [ZIT EET 13310 MOT 790 [IIFEea) | "05s | Iili | 9 | : 23i ld Range pw & | 13310 Tine 18310 Escwbxit 1 FiHelp | CriI-PeEAit peaks | Printout of Sample Table : 63291350 Inserted in System Table of : group 1 0iZaciw InttiaTlrDwaracisefrtn 111 111...0000000000 - ii$v%5 sSttaannadaaVrraadni: 111 11i..000000000 - $55830 355 SSStttaaannndddsaarrraaasss 111 111...000000000 - 1$03 57 SSiiaannddaarraays 11 11.000000 i138 standards 1 1.000 00793-- 1995-08-22 09:05 OutPut of : 950822A1 Software : version 6.1 1990,93 operator + Dow Date of the Analysis : 1995-08-22 06:49 Analysis Pile Name : C:\SKALAR\DATAVHWIDATA\SERUN\S50822A1 000 tasks (2 VOT 2075.1 HoT 329.135 PE FCalluiobrriadteio1n.5order = Inverse Logarithm stope Py resut = 10 [x2-25e2l] xh 5" == lcSloorpreescotfedtvvheaalleiuleoocfftrotBdheee aaz== 0T.o0l0ao0o0r0s 0 lien CFaltufobrriadteioLn order = 2 Correlation : r = 0.99917 Res=ua2l+xt? +a%xl +a0 aa2zi01=I 0TS.ol0lo0ooo0so0rz0rr Sampler HSTuyampmbpelre Time :H+h sa1000 50 sec. AWarsh tTiinnee 1: 1120 sseecc.. nSTeaPekedecliaeulpHeight |: N7so0inneglmae. sBamiplelan s Diluter ReadnsiielleauudnlttpeiiiooennHeiFVgaoclhtutomre i::: i8120.05 mm ml. Dilution runs 1 URseeprrocfeisle :: No Ler 0600794 1995-08-22 09:05 output of : Fluoride 1.5 SPiagtnhalnumtbyepre :: De3bubbled sDaycsotleomrNumber :: 0Yes dRielsuatmeple i: NNoo 4diiGl oTuhtrpeusthold :: 04095 Window event : Off ss21 ssTTaannddaarrdd :: IIggnnoorree s543 ssTTaannddaarrdd :: IIggnnoorree ss56 ssTTaannddaarrdd :: Igno0r.e150 $s87 ssTTaannddaarrdd :: 00..630000 $5190 ssTtaannddaradr:d: 11..520000 . oDridmeernsi: onInv: erPsPeM Logarithm tstrairgtgerValLuismit :: 1508000 SDUec PSetaAkrtshiagpneore :: 6P0ointeSdec MoNodasureigwniornedow :: 71520 8Sec RFielgteenreration :: NNoo ofourtmplaut:: #H.#8F Fluoride L PSaitgnhalnumtbyepre :: De0bubbled sDeycsotleomrNumber :i 0No dRieLsuatmeple :: NNoo i4iGl oTuhtrpeusthold:: 04095 Window event : Off 950822A1 600 795---- 1995-08-22 09:05 output of : ssi2 ssTTaannddaarrdd :: 00..003105 5s3i ssTTaannddaarrdd :: 00..009600 ss56 ssTTaannddaarrdd :: 00..115200 s578 ssTTaannddaarrdd :: IIggnnoorree 55910 ssTTaannddaarrdd :: IIggnnoorree Doridmeernsi: on2 : PPM sttrairgtgervalLuiemit :: 1580000 DSUec sPteAarktshiagpneore :: P60ointeSedc MeeNadsureigwnionredow :: 17250 Sec FiRelgteenreration :: NNoo ofourtnUplaut:: c#d.:#=8c83 950822A1 600796 1995-08-22 09:05 output of : 950822A1 Fluoride 1.5 Fluoride Page 1 Pos Typ wt FH oie w i5ow0% 51 57 5S5i fSo 585 1315108S25l5o 1ii8oawU0 17 4ou 2Fe ou 3320380W F335I%a 3$J5o4uW 33FH au 3350%uw 3$44 930 aoUw Pr wG4ri2ioaYuu on eon 1dent Ch Result 7 Time Ch Result 7 Time InitialTbrrawicateserh 333 101...440766515 32806385 444000...077076526380 0 a stancaWraasih Standara2 33 3 0l0..o00g85s5 7s6201976 444 000l.l00oz01a5318 SSStttaaannndddaaarrradass4 333 000l..l0129183 111402388335 444 00o...10o1662o73 o SSttaanndadradr7d6 Standards 33 3 000...16332771 1ii9ts5es9s 444 000...z4is4se4ir3z ao sStatnadnedDrararidf1st0 333 1D1..a436z%5 4223010449 44400.0..762670030720 a sSmewmniiWknas2dh 333 0o0..l006s75 23207812312 444 000...000340378428 SSSPPeKk444000-323 333 000..l00038%03 333331368317 444 000...000854773302 0ao sSSeeerxiaz4zi0ed4 33 5 oo0.i0i9s2 d3e7o3ss0 44400o..s00e02:80 0 SSihzDiierade 33 3 o1o.i5z3e3 44452183248 444000..72a9899m48 o0 sSeexxaioWcoaoeszrh 333 o00..a028r15 s4i9us3r26 4440.00.11003033798 aoo SioBBaiixx 333 0o0.i038i0 3s56a43m63e0 444000..i00237m93e05 ao0 pIrsgsaeaeeoriae2sz0 333 lcololorseee e0si8es7ii 444000...00032438441 a rBsazersdezsz rife 33 3 o1ol.lo5or3r02s eeeeasissi 444 000l..o70i9me8ss3 F saseWsaisnh 33 l0o.r0e8 7603E 3271S+ 440.0l0+ o0z5e8sSEES o ,veORnLY runSoPuKtD1wWr0aia0sf-sht1h 3333 0001....001368255510 217733300863361 4444000.0...0s01000038558829 0oa *halas of 1 SS Coo 600 797---- || 0.20511 Calibration curve of SS0G22AL : Fluoride L -------------- Channel 0 Measured: BH Concentration: MINES] Delta: CXR | | / 3 H: t $ 0.005706 | | Order 2 Heasured 500 ri 0.99917 600798" __ | ems t| | | 1 i ; calibration curve of 503221 : Fluoride 1.5 yam y [! I I!| i | | 0.064727 Weasured Order : Inverse Logarithm | 005 s 599 606799" teu date of sown Fluoride tS 1] | 4035 | 1| | | | IT o i 7 ||| i 3 -9 . | " ", u 9 lel a 1 Vl Sa sie A v .: i oh Coortuit Fis | GE pan 6006800" ------------ Rau data of 9508221 Fluoride 15 -- 4085 EXIT MTT TAco)oWYP aoe GYSURWea || |. 1 " | | 1 | | s | | || "5 | & A ToFOoR nAAa "0 2 lo 0 4560 Tine Esc=Exit ! Fi=Help ! Crtl-P=Edit peaks ! 9560 " 000801"