Document e7vewVYKXZno7eMQEXNkmrEde

LHR - 0500 - 373 3M Environmental Laborato! & ARA26-094-8 x3 Final Report- Analytical Study BE Single-dose Intravenous Pharmacokinetic Study of T6246 in Rabbits In-Vivo Study Reference Number: HWI#6329-159 Study Number: Test Substance: AMDT-042095.1 FC-95 (T-6246) Name and Address of Sponsor: 336M7 SGrCoDveDiSvtirseieotn St. Paul, MN 55106 ePA-OTS FEV gRasiyresy Name and Addressof Testing Facility: 3M Environmental Technology & Services 935 Bush Avenue = St. Paul, MN 55106 89 MAeMtDhTo-dM-N1u-m0b,ers TDahonehdrrmRmaeavlnisEnxitoDrnXas2c:t0i0on0oOfrFglaunoircidHealbiydemAenaanlsyzoerfa-LMiovdeirfied v3 hil 2 oF AMDT-M-2-0, EFlxupoarniddaebMleealsounreAmnaelnytzebry Means of an Orion EA940 nO AAMMDDTT-MM.-84.-00,, AExntarlaycstiisonofofFlFulouroirdoecUhseimnigcatlhse fSrkoamlaRraSbebgitmeLnitveerd Flow Analyzer AMDT-M-14-0, with Ton Thermal SEexltercatcitvieonEloefcFtlruoodreide by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum Initiation Date: See attached protocol Author: James D. Johnson Ape proveds By: adlyes DDi.reTctoohfagl Cre Mn om fer fps Completion Date 000081 0f0ac0k6--01 4 23> 10SUMMARY 000000000 Rabbit serum and liver were analyzed for fluorine content at various times after rFaCb-b9i5t.s wDeorseesdoasreedeixnptrreasvseendouasslyFCw-i9t5h aeqsuiinvgalleendtoss.eTohfTe-f6l2uo4r6i,neain0.s1e2ru%msowlaustion of detected after a single dose of 0.12 mg/kg and after 0.60 mg/kg, but not after a 0.012 mg/kg dose at 28 days post dose. The method is not sensitive enough to detect and measure fluoride after the 0.012 mg/kg dose. The half-life of perfluorooctanesulfonate anion was found to be one month or greater in serum and liver. The average liver fluoride ion concentration in Group 5 animals (0.60 mg/kg), at day 14 post dose was 1.48 ppm and for four other animals in the same group sacrificed at day 28 the concentration was 1.23 ppm. Thus, the decrease in concentration in liver from 14 days to 28 days post dose is slow; based on just these two points the half-life is greater than 1 month. 2.0 INTRODUCTION `This study was designed to provide information as to whether the perfluorooctanesulfonate anion does go to the liver and other tissues when the `cmoantceerniatlraitsiaodnmiwniitshtteirmeed aifntaern dionsteraivnesnoeursumdoasen,d alnivdert.o ascertain the change in Ipterisflkunoroowonctfarneosmulsftuodniaetse daonnieonprisevqiuoiutsellyonwgit(h>1ratmsonthtaht).theIthawlaf-sliefxepoecfted that the half-life in rabbits would also be long. Perfluorooctanesulfonate anion is a likely biotransformation product of several fluorochemicals that are to be tested for dermal absorption in other studies. The pharmacokineticsofthe perfluorooctane- sulfonate anion is very relevant to these dermal absorption studies. 3. EST MATERIALS 3.1 Tes3t.,1.C1onAtnraolly,tiacnadl RReeffeerreennccee SSuubbssttaannccee:s aFnCd-9M5a,trloitce1s61 or 171. They are equivalent. 3.1.2 Analytical Reference Matrix: Bovine liver and bovine serum 3.1.3 Analytical Control Substance: None 3.1.4 Analytical Control Matrix: Bovine liver and bovine serum 000082 000003, 3.2 Source of Materials: 3M ICP/PCP Division For FC-95. Bovine liver from grocery store, bovine serum from Sigma Chemical Company 3.3. Purity and Strengthof Reference Substance: Responsibility of Sponsor. 3.4 Stability of Reference Substance: To be determined by Sponsor. 3.5 Storage Conditions for Test Materials: Room temperature for FC-95. For biological samples, the storage is -20+10 C. 3.6 Disposition of Specimens: Biological tissues and fluids will be retained per GLP Regulation for the time period required for studies longer than 28 days. 4,0 EXPERIMENTAL-Overview Serum and tissues from animals dosed as described (HW1#6329-159), were available for analysis for fluorine compounds. Since perfluorooctanesulfonate anion is not biotransformed, the analysis was accomplished with combustion and subsequent analysis for fluorine. The fluorine data are related directly to perfluorooctanesulfonate concentration. The fluorine analysis of serum collected at different time intervals after dosing provides data which can be interpreted pharmacokinetically. Additional analysis of liver samples provides evidence that the perfluorooctanesulfonate anion is still present at 28 days. 5.0 EXPERIMENTAL - Methods 5.1 AMDT-M-1-0, Thermal Extraction of Fluoride by Meansof a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver 5.2 AMDT-M-2-0, Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer 5.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver 5.4 AMDT-M-8-0, Analysis of Fluoride Using the Skalar Segmented Flow Analyzer with lon Selective Electrode 5.5 AMDT-M-14-0, Thermal Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum 000083 000603 3 6.0 DATA ANALYSIS Raw data are shown in Table I (Appendix 9.11). Doses are adjusted for the FC-95 equivalents. The administered dose was T-6246 which is a water solution of 0.12% FC-95. Serum concentrations are obviously not different than the control concentrations for the 0.006 and 0.012 mg/kg doses. The concentration in serum is plotted for Group 4 and Group 5 (see pages 140-141). For Group 5 (0.60 mg/kg dose), if the average concentration is fit to a least square regression Y=aeTM, 0.95, a=1.62, k=-1.099 x 10, k=0.693 overt,,, and t,,=26 days. Similarly, t,,=48 days for Group 4. Thus, if Perfluorooctanesulfonate ion is formed from biotransformation, a large portion of the amount formed will be present in the serum at 28 days. `The average liver fluoride ion concentration in Group 5 animals, at day 14 post dose was 1.48 ppm and for four other animals in the same group sacrificed at day 28 the concentration was 1.23 ppm. Thus, the decrease in concentration in liver from 14 days to 28 days post dose is slow; based on just these two points the half-life is greater than 1 month. Overall, for all time periods the serum and liver levels are as expected for the i1n2t0rauvge/nkogussudgogsees.tsTthheatabwihleitny pteordfeltueocrtoolcevtealnsesiunllfiovneartaeftiesrthaen mianrtkrearv,endoeursmdaolse of absorption studies wil be quite sensitive. 6.1 Circumstances That May Affect the Quality of the Data: These values for concentration are from combustion analyses of the biological material. The recoveries are based on spiking the biological material with known amounts of FC-95. The fluorine is measured by selective ion electrode and the Orion meter is calibrated over a 5 point range. However,ifthere is a different response at different concentrations of FC-95 other than the point used as a calibration check, there could be a bias at those concentrations even though the amount of fluorine is being measured accurately. This can come from other variables in the method such as the efficiencyof combustion. Thus, the values reported for concentration are not absolute values. The values are certainly sufficient to make the point that the perfluorooctanesulfonate anion is present at 28 days in liver and serum, that the half- life is on the order ofa month or longer, and that the method can detect perfluorooctanesulfonate in serum after a dose somewhere between 0.012 and 0.12 mg/kg. This method allows sufficient sensitivity for dermal absorption studies wsuhlefroenasteomaenoiofn.the applied fluorochemical is biotransformed to perfluorooctane- 000084 00eeoa, CONCLUSION The half-life of perfluorooctanesulfonate in liver and serum after an intravenous ddoetseectisedonbey mthoentchomobrusgrteiaotnermeinthraobdbiatns.d sTuhbesepqeurefnltuofrlouoocrtiadneeasnuallfyosniasteata2n8iodnacysanafbteer cither a 0.12 or a 0.60 mg/kg intravenous dose. The 0.012 mg/kg intravenous dose dprooebsanbolty pbreovaibdlee tsoufdfeitceicetntafdlousoreidseomioenwhfeorrethibsetmweetheond0..0T1h2eanmdet0h.o1d2 would mg/kg. Analysis of fluorine at 28 days in liver and serum provides a marker for dermal apebrsfolrupotrioooncftoarnedseurlmfaolnlayteapapnliioend. fluorochemicals that are biotransformed to 8.0 MAINTENANCOEF 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. 90 APPENDICES 9.1 Protocol and Amendments 9.1.1 Protocol and Final Report: HWI#6329-159 "Single Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits" (Protocol type TP8084.PK for dosing of animals, tissue collection, etc.) 9.1.2 Analytical protocol AMDT-042095.1 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 000085 000000000735 9.9 Test Methods: See Protocol AMDT-042095.1 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; ug F in whole liver as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. 9.11.3 Summary and raw data; ppm F' in 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. 000086 000008 6 9.1.1 Protocol and Final Report: HWI#6329-159 "Single Dose Intravenous Pharmacokinetic Studyof T-6246 in Rabbits" (Protocol type TP8084.PK for dosing of animals, tissue collection, etc.) 000087 000007 @ HAZLETON SABINA Sponsor: 3M St. Paul, Minnesota FINAL REPORT + ComnNG Corsi, 7eEosCSEe,iad {j se3Pp 14741819959\51 So. dMiA s Study Title: Single-Dose Study IonftrTa-v6e2n4o6usinPhRaarbmbaictoskinetic Author: Steven M. Glaza Study Completion Date: September 14, 1995 Performing Laboratory: Ha33z0l1etoKninsWmiasnconBsoiunl,evaIrncd. Madison, Wisconsin 53704 Laboratory Project Identification: HAT 6329-159 Phone soe2uv.eert EXPRESS-MAIL DELIVERY. Page 1 of 38 000608 Fax 508.241.2227 3301 KINSMAN BLVD MADISON. wi 53704 000088 Page 2 of 38 HWI 6329-159 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit of Hazleton Wisconsin, Inc., in accordance with the Food and Drug Administration (FDA) Good Laboratory Practice Regulations, 21 CFR 58.35 (b) (6) (7). The following inspections were conducted and findings reported to the Study Director and management. Written status reports of inspections and findings are issued to Hazleton management monthly according to standard operating procedures. From Inspection ToDates 03/22/95 04/10/95 04/27/95 07/05/95 09/12/95 03/22/95 04/10/95 04/27/95 07/10/95 09/12/95 Phase Protocol Review Protocol Amendment Animal Observation Data/Report Review Report Rereview Date StRuedpyortDeidrecttoor 03/22/95 04/10/95 04/27/95 07/10/95 09/12/95 MaDantaegemteont 03/10/95 05/10/95 05/10/95 08/10/95 10/10/95 A) AZ Represefititive, Quality Assurance Unit 14.95 Date 000089 000009 Page 3 of 38 STUDY IDENTIFICATION Single-Dose Study IonftrTa-v6e2n4o6usinPhRaarbmbaictoskinetic HHI 6329-159 Test Material Sponsor Sponsor's Representative Study Director Study Location StSutduydyTimIentiatbilaetion Date EIxnp-eTriifmeenEtnadl Da(tIen-1ife) Start Date SEtxupedryinCeonmtpalletiToenrmiDnaatetion Date T-6246 TMonxicology Service 3MMeCdeinctaelr, DeBlpdagr.tme2n2t0-2-02 PSt..0. PaBuolx, 33MN22055133-3220 JMnohn L. Butenhoff, PhD ToMxeidcioclaolgyDeSpearrvtimceent 3P.M0.CeBnotexr,332B2ld0g. 20-26-02 S(t6.12)Pau7l3,3-1M9N62 55133-3220 SHtaezvleentonM.WiGslcaoznasin, Inc. MPa.0d.isoBno,x W7I54553707-7545 (508) 241-7292 3H3a0z1letKoinnsmWainscoBnosuilne,varIdnc. Madison, WI 53704 AMparriclh 43,0, 19199595 MSaeypte1,mbe1r99514, 1995 September 14, 1395 000090 000010 Acute Toxicology SSttuedvyenDMi.recGtloarza Manager FSrtaundcyisCoo(rBduid)natWo.rMcDonald PIant-rTiicfieaSuPpaedrgvhiasmor Rose M. Bridge Report Supervisor Toxicology Support Kathy Myers Manager CSaulpveirnvisL.orHorton Page 4 of 38 KEY PERSONNEL HW 6329-159 Quality Assurance MSahneargreyr R. W. Petsel aboratory Animal Medicine DCiipnldoymaJt.e,CarAyC,LAMDVM Supervisor Anatomical Pathology Thomas E. Anatomical Palmer, PhD Pathologist Jack Serfort/ DSeubpoerravhisoLr.s Pirkel Necropsy Anne Mosher SPautpheorlvoigsyorData 000091 000011 Page 5 of 38 CONTENTS Quality Assurance Statement Study Identification Key Personnel Summary Objective Regulatory Compliance Test and Control Materials Test System Procedures Results Discussion Signature Pathology Report Table 1 Individual Body Weights (9) 2 Individual Clinical Signs 3 Individual Pathology Comments 4 Individual Animal Tissue Weights and Bile Volumes Appendix A Pretece] Protocol Protocol Mmencmant Mo. Deviations TP8084.PK 1 HWI 6329-159 Page 2 3 4 6 8 8 8 9 10 13 13 14 15 16 18 21 Ze 25 Zz26 27 000092: 000612o Page 6 of 38 HIT 6329-159 SUMMARY Tahdimsinissttuedryedwasbydoanesintogleassienstsravtheenoulseveilnjoecftisoynstetomicrabebxiptoss.ure of T-6246 when The the Hsrtau:dy(NZwWas)SPcFondsutcrtaeidn ufosringeacfhourtremaaltemenatndgrfoouupr afsemafloelloawcsc:limated rabbits of Group 1 (Control) 32 ((LMoewd)ium) 45 ((HHiigghh)) Test Materia Sterile water TT--66224466 TT--66224466 Dosnoe/kLg)evel 0 105 150000 MNalu es m ofbAFnee immsalr less 4 4 4 4 4a 44 a 4 * pOnoessiabnlimealbroskaecnribfaicckedanadfterrepltarceeadtmweintthduaenotthoera male aninal. tThhee iannidmiaclastedrecdeosievedlevaelsinignlteo tihnetrmaavregnionuasl ienajrectvieoinn dose volume was 0.5 ml/kg of body weight for Groups ooff tthhee rteisgthtmaetaer.rialTheat 1 through 4 and was Da0an.yd75t28hm.el/krgemafionrinGgrouapnim5a. ls "T(wtowoaniamnailnsa/lsse/xs/edxo/sdeoselevleelvelw)erewerseacrsiafcirciefdiceodn Day on 15 4Clihnoiucrasl aofbtesrervianttiroanvsenowuesreicnojnecdtuicotne.d prAdeddiotsieonaanld actlinaipcparloxoibmsaetrevlayti0o.5n,s twice a day mortality checks were conducted daily thereafter until tha 2a,nd and oasncdhmDeiadnyuilset-dr7atfsioaorcnrirfa(inDcdaeoymii1z)n,atteiraovnandl patu(rDptaohysee1s5s,chorebdeufDloaeryde28st)ae.sctrifoBirocdeyconwitenritoeglrhvtasmlatwee(rrDieaaylde15teromrined eDaary v2e8)i.n (Alefbtlooedar)samopflethe(aapnpirmoaxlismatately4-,4 8n-l,) w1a2s-, co2l4-l,ectaendd f48r-ohmoaursmarginal pcInooslltae-dcidtniejtideocnto,inonDa,atystahned15toniamnedDayo2f28.tfhoerAnstchehaepdpaurnloiexmdailmsastaecsrc4ih-femidlcuelebdl(oDoafdyor1s5asmapcolrreifDiawcyaes28ao)nl,soDay 28. caepnptrroixfiumgaetde,ly se20pamralteodf Sponsor." On Day 15 or bilnotoodswearsunobantdainceedlluflraorm efarcahctiaonnism,al.andAlslentsamfopltshewere 28, the animals were anesthetized with sodium capoeblnbltreoecbvtaieradbt.ietdalgT,rhoesbswlheodnleecvriaolpisvteyhre,expaboimslitene,artiioaornnd vwebanosathcnoaktviad,dnoenyaesn,d frehooxwmseavneegaruc,ihnatatnieisdms.aulesAwnweerree cSpoolnlseocrt.ed, weighed (volume only determined for bile), and sent frozen to the 000093 000613 Page 7 of 38 HHI 6329-159 cIhnatnrgaevsenoiuns boidnyjewcetiigohntogfaiTn-.6246Alldidaninmoatlsresauplpteariendancylintiecsatllmyatenrorimaall-related atnhirmoaulgshoutthatthewersetudsyacrwiifthicetdheonexcDeaypstio2n anodf o1,nereGsrpoeucpti4vealnyd, ondeueGrtooupin5jumrayle (possible broken animals appeared cblaicknsi)c.allyThensoremaalnitmahlrsougwheoruet rethpelacsetdudya.nd the replacement 000094 000011 Page 8 of 38 HHI 6329-159 OBJECTIVE The objective of this study was to assess the level of systemic exposure to the test material, T-6246, when administered as a single intravenous injection to rabbits. REGULATORY COMPLIANCE This study was conducted in accordance with the U.S. Food and Drug Administration's Good Laboratory Practice Regulations for Nonclinical Laboratory Studies, 21 CFR 58, with the exception that analysis of the test mixtures for concentration, homogeneity/solubility, and stability was not conducted. All procedures used in this study were in compliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study did not unnecessarily duplicate any previous work. TEST AND CONTROL MATERIALS Identification a clear, colorless liquid The test material was identified as T-6246 and described as a clear, colorless liquid. The control material was Sterile Water for Injection, USP (Abbott Laboratories, Lot No. 86-748-0M-02; Exp. April 1, 1996), and was described as Purity and Stability The Sponsor assumes responsibility for test material purity and stability determinations (including under test conditions). Analysis of the test material mixtures for concentration, homogeneity/solubility, and stability was not conducted or requested by the Sponsor. The purity and stability of the control material were considered to be adequate for the purposes of this study. Storage and Retention Tshteoretdestrefmartiegreiraaltedw.as stored at room temperature. Any unused test material was The control returned to material was the Sponsor after completion of all in-life testing according to Hazleton Wisconsin (HWI) Standard Operating Procedure (SOP). Any remaining vehicle may be used for other testing and will not be discarded after issuance of the final report. 000095 000015 Page 9 of 38 safety Precautions HHT 6329-159 aTnhde tpeoslticiaensd. control material handling procedures were according to HHI SOPs TEST SYSTEM Test Animal AKdaullatmazaolob,inoMicrhaibgbaintsonofMatrhcehH8r,a:1(9N9Z5)SaPnFd msatirnatianinweedreatrecteheiveHdazlfertoomnHRWPi,scoInnsci.,n facility at 3301 Kinsman Boulevard, Madison, Wisconsin. Housing ADufrtienrgreaccecilpitm,atitohne aanndimatlhsrowuegrheoutactchleimastteuddy,forthea apneirmiaoldsowferaet lienadsitvid7uadlaylsy. hcmoaouinsntetrdaoilnlienda stqcueramerpetener-rabsto.utrteomEonfvsitra1oi9nnmleetnsotsa2l3stCec,eolntarcoarlgeselsatfioivrnetthehemupneiardnaiitmtauylreo-frooa5mn0d%w2ehru0em%i,dsiettayn-dtoa t1h2e-sheourcoTndiigthito/n1s2-heoxuisrtedda,rkthTeiyghwteirneg dcoyccluem.enteIdn adverse effect on the study outcome. canadsescownhseirdeerevdaritaotihoanvse hfardomno AnimalDiet LThaeboraantiomraylsRawbebrietprDoievtideHdF a#5c3c2e6s,s tPoHIwaFteeedrs,adInlci.bitTuhmeanfdeeda miesasruoruetdinealmyount of cwaoennratelaymznioendaknnbtoysw.nthecoSmnaatmnapumlfieanscatnoutfrsetrhine fwtorhaetenruftereadirteioorpneawrlaitoedcriocmaaptlolnyelnetvasenlaslaynzdtehdaetnvbiywrouoHlnHdmI.enhtaavTlehere interfered with or affected the results of the study. Selection of Test Animals wTehreeapnliamcaelds wienrteo sitduednytifgireodupsbyusaniinmgala nsutmrbaetirfiaendd bcoodryreswpeoingdhitngranedaormitzagatiaonnd wperiogghratm.variTahteiornandoofmitzhaetiaonnimablosdyfwoerigehatcsh wgerroeupdeotferemaicnhedsexon sDealyact-e7d. fTohrethe bsotduydywediidghtnsotfoerxceeeadch+2grosutpandoafrdeacdhevisaextiownesreofnotthestmaetainstiweciaglhlty, diafndfertehnetmeaatn the 5% probability level. 000096 000016 Page 10 of 38 Study Desian Ainnitmoaltshewefioglhlionwgingfrosmtud2y,52g3routpos:3,276 g at initiation Group Test/Control Dose Level VoDlosueme Material naka) (nl/ka) 21 ((LCoown)trol) StTe-ri6l2e46water 05 00.55 34 ((HMiegdhi)um) 11--66224466 10100 00:55 5 (High) T-6246 500 0.75 HUI 6329-159 of treatment were placed MNaulmbeesr of FAneimmaalless 44 44 4a 44 a 4 * Obnreokeannimbaalcksaancdrifriecpeldaceadftewrithtreaantomtehnetr dmuaeletoaniamaplo.ssible a Trweomaianniinmgalsa/nsienxal/sdos(etwolevaenlimwaelrse/sesxa/cdroisfeiceldeveoln)Dwaeyre15. The sacrificed on Day 28. Justification for Species Selection cHhiositcoericbaelclayu,sethoef tNheew lZaeragleandamWohuintteofalbbiancokgrroaubnbditinhafsormbaeteniontheonantihmiasl spoefcies. PROCEDURES Dose Preparation and Administration The test material concentration for weaaschdidlosuetedlevweilth insteGrrioluepswa2tetro 5t.o aAcnhieivnedivaidsupaelcifdiocse of eonachitsresbpoedcytiwveeighttestonstohleutidoany oorf ctornetartomlentw.as cThaelcurleastpeedctifvoer eteascth asnoilmuatlionbasweads aeadrminoivesrteraepdprobxyimiantterlayven2o2ustoi6n0jecsteicoonndsi.ntoThteheprmearpgairneadl teesatr vseoilnutiofonsthewerreight rsetmoareidninagt rteosotm tsoelmupteiroantsurweeruentidliscaadrmdiendi.stered. After administration, any Reason for Route of Administration Intravenous injection is an acceptable route to assess systemic exposure. 000097 000017 Page 11 of 38 Observations of Animals HH 6329-159 C4lihnoiucrasl aofbtseerrvianttiroanvsenowuesreicnojnedctuicotne.d prAedddiotsieonaanld catlinaipcparloxiombasteerlvyati0o.n5,s a2,ndand tswcihceedulaeddaysacmroirftiacleityintcehrevcakls w(eDraey cIo5ndoructDeady d28a)i.ly thereafter until the tBeosdty owreigchotnstrow]erematdeertiearlminaeddminonistDrayati-o7nfo(rDayran1)d.omizAadtdiiotinonpaulrpobsoedsy waenidghbtesforweere dunestcehremdiunleeddatsacthreifisccehsedu(lwehdensasucrrviifviacle eixncteerevdaeld 1(Ddaayy)1.5 or Day 28) or at sample Collections A(lebfltoodears)ampolfeth(eapapnriomxailmsatealty4-&,mL8)-,wa1s2-,col2l4-e,cteadnd f4r8on-hotuhresmaprogsitn-ailnjeecatriovne,in aanndd 2o2n Dfaoyr 8t.he Aannimaaplpsroxsicmhaetdeule4d'nLforblosoadcrisfaimcpeleonwasDayals28o. colInlecatdedditioonn,Dayast t15he tviennea ocfavanecorfopesayc,h aanpipmraolx.imaAtlellysa20mplmelsowferbeloosdtowraesd oatbtarionoemdtefmropmeratthuerep,ostaendrior wtehreen tcheenntrisftuogreedd, insepaarfarteeedzerintsoetsetorummaianntdaicnellaultaermpefrraatctuiroenso.f -T2h0esCe+1S0amCples until shipped to the Sponsor. Pathology Tnheecroapnsiimeadls isnactrhiefiscaemde monanDnaeyrsas1 atnhde 2andiumaelstosuarpvpiavrienngt btoroktehne bsacchkesduwleerde seaxcarmiifniacteisonwiatnhd tahney eaxbcneoprtmiaolnititehsatwetrheeyrerceocredievde.d an abbreviated gross necropsy gOnrouDpayas15s,igntmheentfirrsatndotmwoizaatniiomna)ls/wseerxe aasnseisgtnheedtizteodeawcihthdossoediulmeveplent(obbaasrebditoanl the a(nvdiaexisnajnegcutiinoanteidn. thAenmaarbgbirneavliateeadr gverions)s, neblcerdopsvyiaexthaeminpaotsitoenriowrasvennoat cdaovnae,, ehaocwhevearn,imatlisswueerse wceorlelecctoeldl,ectweedi.ghedThe(vwohloulmee olnilveyr,detbeilrem,ineadndfborothbilkei)d,neyasndfrom Aifmtmeerdiattieslsyue/pblialceedcoilnleactfiroene,zerthesetanitmoalmsainwtearien diasctaermdpeedr.aturTeheofrem-2a0inCing10twCo. athneinasalmse/semxa/ndnoesreaslevtehle waenriemalasnesstahcertiifziecde,d bonledD,ayan1d5. exsanguinated on Day 28 in 000098 00001S Page 12 of 38 Shipment of Tissues HHI 6329-159 After completion of fractions), livers, the in-life phase, the bile, and kidneys were blood samples (serum and sent frozen (on dry ice) cellular to the Sponsor (James D. Paul, MN, 55106), Johnson, 3M E.E. along with their & P.C., Bldg. corresponding 2-3E-09, 935 Bush Avenue, weights or volumes. The St. SHHpIonsdoores insotresapcocenpstiblaeny forrespthoensriebtielnittiyonforandthedisapnoasliytsiiosn ofofthtehe sasmampplleess. collected in this study nor are these results presented in this report. Statistical Analyses No statistical analyses were required by the protocol. ocation of Raw Data, Records, and Final Report rTheetairnaewddaitna,therecaorrcdhsi,vesandofaHnNIoriigninaaclcorsdiagnnceed copy with of the final HWI SOP. report will be 0000339 000012 Page 13 of 38 HUT 6329-159 RESULTS Body Weights bIonddyiviwdeuiaglhtbogdayinwediugrhitnsg atrhee isntudTya.ble 1. There was no meaningful effect on Clinical Observations tIhnrdoiuvigdhuoault ctlhienisctauldyswiigntsh tahree ifnolTlaobwlieng2.exceAlpltioannsi:mals appeared normal hOnaeveGrionujpure4dmailtse b(aNoc.k oFn541D0a6y) 2tranedatweadswistahcriTf-i6c2e4d6, atnec1r00opsmige/dk,g aanpdpeared to trherpoluagcheodutwitthheNos.tudFy5.4099. The replacement animal appeared normal OhnaeveGrionujpure5dmailtse b(aNoc.k oFn540t7h6e)datyreaotfedtrewaitthmenTt-62a4n6d awtas50s0acmrgi/fikcgeda,ppeared to nnoercmraolpsitehdr,ouagnhdoutreptlheacesdtuwdyi.th No. F54112. The replacement animal appeared Pathology aInnidniavlidutailssuaenimwaelighptasthoalndogybilceommveonltusmesarearepreinsenTtaebdle i4n. TabTlhee 3n.ecroIpnsdyiviodfuaalnimal dNoi.ffuFs5e4l1y06darrekvearlede.d the In asnkiemlae]talNo.musFc5l4e076s,urrtoheundsiknelgettahle lumbar muscle spinal cord surrounding was broetmhainhiinngd aTinnibmsalsandsurtvhievesdacrtaol thsepiinralscchoerddulewdassdaicfrfiufsiecleyadndarwkerreed.notTheexamined grossly, although tissues were saved. Page 15 contains a pathology report by the study pathologist. DISCUSSION TahlebinToeverlabboiftssywshteenmicadmeixnpiossutreeredofasT-6a24s6inwgalse eivnatlruaavteendousinimnajleectiaonnd aftemalleevels oafny5,of10t,he 10a0n,imaalnsd 5f0o0llmogw/ikngg. admThienriestwrearteionnooftestthismamtaetreiraila-lr.elated effects in 000100 000020 Steven M. Glaza Study Director Acute Toxicology NN Page 14 of 38 SIGNATURE HHI 6329-159 AAS Date 000101 000021 Page 15 of 38 HHI 6329-159 PATHOLOGY REPORT There were two male rabbits sacrificed on Day 1 or Day 2 and necropsied. At necropsy, the skeletal muscle surrounding the lumbar spinal cord in Animal No. F54106 (100 mg/kg of body weight) was diffusely dark red. In Animal No. F54076 (500 mg/kg of body weight), the skeletal muscle surrounding both hind Timbs and the sacral spinal cord was diffusely dark red. The findings in these animals are indicative of injury of undetermined etiology to the caudal region of the back. The liver, bile, and both kidneys from these animals were collected as required by protocol. After necropsy, the animals were discarded. "4 C a orr . Thoma$.F. mer, "PhD Pathologist (6329-159. 1hm) 051295 I Date 000102 000022 Page 16 of 38 HWI 6329-159 Individual TaBboldey W1eights (g) . Random- Sex AnNibmealr i(zaQtuio)n InQiatlilal Group Control) - Sterile Water for DadsTerminaDZ Injection (0 mg/kg' Male F54105 F54075 F54081 F54107 2,603 2,531 2,313 2,544 2,761 2,539 2,526 2,731 2,933 2,703 + - 2,874 3,056 Female F54104 F54086 F54074 F54085 2,665 2,526 2,540 2,553 2,890 2,632 2,703 2,667 3,130 2,775 - - 2,990 3,014 Grow 2 - T-6246 (5 ma/ka Male F54083 F54100 F54077 F54094 2,381 2,675 2,459 2,466 2,598 2,869 2,592 2,601 2,538 3,030 - - 2,859 2,916 Female F54091 F54108 F54102 F54097 2,717 2,412 2,690 2,439 2,785 2,622 2,839 2,533 2,996 2,790 - - 3,290 2,928 Grow 3 = T6246 (10 maka) Male F54071 F54093 F54095 F54101 2,487 2,652 2,662 2,622 2,523 2,776 2,772 2,642 2,709 2,993 - - 3,126 3,162 Female F54110 F54096 F54090 F54078 2,719 2,772 2,542 2,710 2,802 2,914 2,740 2,842 2,959 3,022 = od = 3,022 3,288 - Not required. 000103 000023 Sex Male Female NAunimmbaelr F54088 FF5544100869 FFS544008979 FF5544009820 FS544l1l0e9 Page 17 of 38 HAT 6329-159 IndivTiadbulael 1Bo(dCyonWteiinugehdt)s (g) Randoni(zDaatyio-n7 InDiatyia1l DavisTerningDay 28 Group 4 - 1-6246 (100 ma/ka 2,494 2,582 2,771 - 22,,657077 2,833 2,662 2,803 - - 2,911 22,,356708 2,26,67564 3,0-04 3,0-60 22,,449080 22,,652420 22,,863657 -- 22,,75796 22,,963537 - 32,,299769 Group 5 - T-6246 (500 ma/ka Male FF5544017163 22,,665657 22,,677583 2,8-9 -- FF54S082 22,,574374 22,,87483 -- 33,,222186 FSl12 2,99 3,276 3,530 - Female F54098 FS408 22,,556150 22,,660709 22,,88251 -- FFa0l7l9s 22,,444628 22,,562161 -- 32,,090529 Mot required. 3 ApnriomgarlamNof.orFu5s4e106inwatsheorstiugdiynalalndy wsaeslecttreeadtebdy. theThirsandanoimmiazlatiwoans sNoa.criF5f4i0c9e9d. on Day 2 due to a broken back and was replaced with ) wNeuimgbhetr wianssutapkeerns.cripted parentheses indicates the day the body ApnriomgarlanNof.orF5us4e076inwatsheorsitguidnyalalnyd wsaeslecttreedatebdy. theThirsandaonimmiazlatiwaosn Nsoa.criFf5i41c1e2d. on Day 1 due to a broken back and was replaced with 000104 000024 Sex Male Female Male Female Page 18 of 38 IndividualTabClleini2cal Signs HHI 6329-159 AnNuimnbaelr _ Observation 0Ho5ur 2(Day 4 7 3tolDsay. Boz Group1 (Control) - Sterile Water for Injection (0 mo/ka F54105 Appeared normal J 4 4 4 4 * FS4075 Appeared normal J J J 4 4 * FS4081 Appeared normal J J J J J v F54107 Appeared normal 4 4 4/4 v F54104 Appeared normal J J J J J * F54086 Appeared normal J 4 J J 4 * FS4074 Appeared normal 4 J J 4 4 v FS4085 Appeared normal 4 J J 4 4 v Group 2 - T-6246 (5 ma/kg F54083 Appeared normal J 4 J 4 J * F54100 Appeared normal 4/4 J J F54077 Appeared normal J JJ 4 v F54094 Appeared normal 4/4 J 4 v F54091 Appeared normal J J J 4 J * FS4108 Appeared normal J 4 J J 4 * FS54102 Appeared normal J 4 J J 4 v FS4097 Appeared normal J 4 4/4 v /* ACnoinmdailtiosnacreixifsitceedd. on Day 15. 000105 000025 Sex Male Female Male Female Aninal Number FS4071 F54093 FS4095 FS4101 F54110 54096 F54090 F54078 F54088 54106 F54089 FS4087 54099" F54080 54092 5409 S414 Page 19 of 38 IndiTvaibdlueal2 C(lCionnitcianluedS)igns HUT 6329-159 Observation= Hour (Day Day 0.5 2 4 2 3fols 16%5 78 Group 3 - T-6246 (10 ma/ka Appeared normal v4 J * Appeared normal 4 J J 4 J B Appeared normal 4 4 J J J v Appeared normal 4 4 4/4 v Appeared normal 4 4 J J J i Appeared normal v4 J J J * Appeared normal 4 J J 4 J v Appeared normal J J 4 4 4 v Group 4 - T-6246 (100 ma/ka) Appeared normal / / J 4 J * ABrpopkeeanredbacnkormal JSLJ LJ - Moribund sacrifice - - J Appeared normal 4 4 J J J v Appeared normal 4 J J J J v Appeared normal 4 4 J J * Appeared normal 4 J J 4 * Appeared normal 4 J J 4 * Appeared normal 4 J 4 J v Appeared normal 4 J 4 4 v /* ACnoinmdailtiosnacreixfisitceedd. on Day 15. +~ RCeopnldaicteimoenntnotaniemvaildenfto.r Animal No. F54106. 000106 000026 Page 20 of 38 Table 2 (Continued) Individual Clinical Signs HWI 6329-159 Animal Sex. Number Observation Hour (Day 1 Day 0.5 2 4 2 3tol5 16tozs Group 5 - T-6246 (500 mg/kg Male F54076 . ABrpopkeeanredbacnkormal Moribund sacrifice 7-oJ-tv --t F54113 Appeared normal 77 v4 7 * F54082 Appeared normal Vv 7 v 7 7 v F54111 Appeared normal Jv 7 J v J v/ F54112 Appeared normal J4 7 v 7 * Female F54098 Appeared normal 7 J 7 Vv 7 * F54084 Appeared normal vJ7v v/ * F54079 Appeared normal 47 vv v J F54116 Appeared normal 4v7J v v * Animal sacrificed on Day 15. / Condition existed. - Condition not evident. t Condition evident at time of 12-hour bleeding interval. a Replacement animal for Animal No. F54076. 000107 000027 Page 21 of 38 Individual Table 3 Pathology Comments HN 6329-159 Animal Number Sex T Died eSs alcaryift iced Necropsy Observation Group 4 - T-6246 (100 ma/ka F54106 M - 2 Tlhuembasrkelseptianlal mucosrcdle issudrirfofuunsdeilnyg the dark red. F54076 " Group - T-6246 (500 ma/kq - 1 The skeletal muscle surrounding bcootrhd hiisnddifTfiumsbeslyanddarskacrraeld. spinal 000108 00002. 8 Page 22 of 38 HHT 6329-159 Individual Aninal TissTuaebleWei4ghts and Bile Volumes Animal Sacrifice Weight (gq Bile Sex Mumber _D _ ay liver Kidneys Volume (mL Group1 (Control) - Sterile Water for Injection (0 ma/ka! Male F54105 15 86.35 14.92 0.6 F54075 15 60.25 15.97 0.1 F54081 28 80.597 14.764 1.2 54107 28 80.649 15.620 0.4 Female F54104 15 98.40 16.03 0.7 F54086 15 77.30 15.23 0.7 F54074 28 82.895 18.793 16 54085 28 80.950 14.925 13 Group 2 - T-6246 (5 ma/ka Male F083 15 73.15 14.87 07 F54100 15 77.90 17.62 0.7 F54077 28 75.699 17.864 0.3 54094 28 78.689 17.213 0.8 Female F54091 15 84.90 14.92 16 F54108 15 79.90 15.40 0.4 54102 28 101.494 18.308 17 F54097 28 73.620 16.214 Ls 000109 000029 Sex Male Female Page 23 of 38 HT 6329-159 Individual AnimaTlablTeiss4ue(CWoenitgihntused)and Bile Volumes Animal Sacrifice Number __Day Weight (q Liver~~ Kidneys Bile Volume (mL Group 3 - T-6246 (10 ma/ka F54071 15 77.10 15.88 0.5 F54093 15 81.40 15.50 0.8 F54095 28 85.504 15.561 1.2 54101 28 85.668 16.120 1.0 F54110 15 83.68 15.79 1.4 F54096 15 78.10 18.70 11 F54090 28 76.220 15.878 2.1 F54078 28 83.460 18.130 1.0 Group 4 - T-6246 (100 ma/kg! Male F54088 15 84.26 16.98 0.4 F54106 2 82.301 16.550 2.0 54089 28 71.79% 16.434 1.0 F54087 28 83.710 18.591 0.5 54099" 15 82.143 16.038 0.5 Female F54080 15 75.42 18.32 1.2 F54092 15 78.36 16.68 - F54103 28 73.958 16.915 1.6 F54114 28 80.966 19.404 2.0 2 Replacement animal for Animal No. F54106. * This animal had no gallbladder, so no bile was collected. 000110 000030 Sex Male Female Page 24 of 38 HAT 6329-159 Individual AnimaTlablTeiss4ue(CWoenitgihntuesd)and Bile Volumes Animal Sacrifice Number __Day eig(h a) t Liver Kidneys Bile Volume (mL Group 5 - T-6246 (500 ma/kq F54076 1 78.088 14.695 2.0 F54113 15 91.52 16.57 1.0 F54082 28 88.965 19.431 1.6 F54111 28 83.823 19.405 1.5 Fsa112* 15 97.824 21.069 1.4 F54098 15 78.95 14.72 1.2 F54084 15 66.42 13.18 0.8 F54079 28 83.117 14.785 1.8 F54116 2 71.962 15.793 2.0 3 Replacement animal for Animal No. F54076. 000111 00071 Page 25 of 38 APPENDIX A Protocol Deviations Protocol TP8084.PK Protocol Amendment No. 1 HWI 6329-159 000112 000032 - Page 26 of 38 Protocol Deviations WI 6329-159 Protoco PC.ageDos6,ing7. PrEoxcpeedruirmeesn,tal(1)Design, DionsjiencgtioRnoutien.to tIhnetrmaavregnionuasl ear vaepipnrooxfimattheelyrig3h0ttoear60ovseerconds. Actual Procedure TGhreoupratefeomfalie'nje(cNot.ionF54f0o9r8)one wtaisme22forsecdoonsdes.admAilnsios,tratthieon fNoo.r oFn5e408m7a)lewaasnimianlcor(rGercotulpy4, druercaotridoend othfusdostihengexcaanctnot be determined. Tohuetsceomedevoifattihoensstuadrye. not considered to have had an adverse effect on the 000113 . 600033  HAZLETON RaBISBNTN 0 Tae Page 27 of 38 + CORNING Corp Sponsor: st. pa hiMmm esota PROTOCOL TP8084.EXT study Title: Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits nates March 30, 1995 bertoraing Lsbratory: HHa0adzrloent,EononVaWmiasrsconBssoiunr,avaiInnrsc.e sboratory project taent ification: HWI 6329-159 ) 000114 00003% Page 28 of 38 3 PPa8g0e842 EXT STUDY IDENTIFICATION Single-Dose IonftrTa-v6e2n4o6usinPhRaarbmbaictoskinetic Study HW No. 6329-159 Test Material T-6246 Sponsor Sponsor's Representative B Study Director Study Location TMonxicology Service 3MMeCdeinctaelr, DeBlpdagr.tme2n0t-26-02 SP.t0..PaBuolx, 3W3N22055133-3220 JMohn L. Butenhoff, Pho ToMxeidcioclaolgyDeSpearrvtinceent 3PM.0.CenBtoexr,332B2l0dg. 20-26-02 s(t6.12)Pau7l3,3-1M9N6255133-3220 SHtaezvleentonM. WiGslcaoznasin, Inc. MP.a0d.isoBno,x 7WI54553707-7545 (608) 241-7292 3H3a0z1letKoinnsmWainscoBnosuilne,varIdnc. Madison, WI 53704 ProEpxopseerdimeSnttuadly STtiamrettabDlaete DErxapfetrimReenptoarlt TDaetremination Date MAparyil1, 4,19915995 June 12, 1995 ) 000115 00035 Page 29 of 38 ; PPa8g0e843. EXT 1. SStiundgyle-Dose Intravenous Pharmacokinetic Study in Rabbits 2. PTauodrmpiaonssisesestseretdheaslevaelsionfglesysitnetmriacveneoxupsosuirnejewchteinonthtoe rtaebsbtitmsaterial is 3. TRheiasulasttourdyy Laboratory wCiolmlplibaencceonducted in : accordance with the following Practice Regulations/Standards/Guiidnesl with the Good ecoxncceepnttiroantitohna,t conducted: saonlaulbyisliistyo,f thhoemogteensetitmya,terainadl smtiaxbtiulrietsy wfiolrl not be [[X]] 2C1ondCuFcRt58as(FaOAN)onregulated Study [[']] 4400 CCFFRR 716902" ((EEPPAA--TFSICFAR)A) [[]1 C5(981N)o3h0san(FNion.al)385(0OEC(DJ)apanese MAFF) [1 Motification No. 313 (Japanese MOH) dAWieTrlefcaptrroeorc,eAcdtutrheeRsegsutilnuadtytihoidnsose.sprontoIotncoutlhneneaocrpeeisnsiianorniclomyopfldiutaphnleciecSaptwoeintshoarnythaepnrdAenvsiitmouaudlsy work. 4. QThuealiptryotoAcsoslu,ranscteudy conduct, and the final report will the Quality Assurance Unit in accordance with Hazleton Wbiescaoundsiitned by (HH1) Standard Operating Procedures (SPs) and policies. 5. Test Material A. TI-d6e2n8t5ification 8. P(ThoysibcealdoDcuemsecnrtiepdtioinn the raw data) C. PTuhreitySpanansdorStaasbsiulmiets responsibility for purity and stability tdeesttermmiantaetriioanls/ve(hiincclleudmiinxgtuurned(esr)fefsotr ccoonndcietnitornast)i.on,Sasmoplluebsiliotfy, haodmmoigneinsetirtayt,ionandifstraebqiuleisttyedanbaylytsheesSpwoinlslorb.e tTahkeensebesfaomrpeles (if ttaekremni)natwiiolnl. be sent to the Sponsor after experimental ! 0. Storage Room temperature 000116 000036 Page 30 of 38 ; . Roserue Samples ite TPBO84. EXT Reserve sample(s) of each batch/lot of test and control materials will be taken for this study. F. fetention HTNhIe tiensta fanrdeezceonrtrsoelt mtaotmeraiianltairnesearvteempsearmaptluesrewiolfl -b2e0Cst+o1r0edCat for 10 years 10 years for per HWI SOP. The, Sponsor will disposition in accordance with be contacted after the appropriate regulatory Good Laboratory Practices. 6. Safety precautions Acnoympluentuisoend otefstthematienr-ilailfewiplhlasebeofretthuersnetdudtyo. the Sponsor after 6. As Contral rMeaqtueirriedalby HWI SOPs and policies A Lentification 5. Sterile Phusica) water for inj guscription ec t io n Clear, colorless liquid 0. storsae C. Purity and Stability The purity and stability of this USP grade material is considered adequate for the purposes of thisstudy. Refrigerated . Reserve sumles . F. See Section etention 5. E. Reserve Samples 6. Safety precautions Any remaining control material may be used for other testing and will not be discarded after issuance of the final report. As required by HWI SOPs and policies 7. Experimental Design A pais (0) sspteceies R 000117 ) (2) Strain/Source Hra: (NZW)SPF/HRP, Inc. 000037 Page 31 of 38 y (3) AAgdeultst Initiation (4) 2Me5atn0t3.at5 Ikngitiation (5) f20umbmearlesandandSex20 females TPP8a0g8e4.5 EXT (6) IIdnednitviidfuiaclatniuomnbered ear tag (7) Husbandry (a) HIonudsiivnigdually, in screen-botton stainless steel cages (heavy gauge) (6) AFomoedasured (PHI Feeds, amIoncu.n)t. ofThLeaboforoadtoriys RraobubtiitnelDyietanHaFlyz#e5d326by etnhevimraonmuefnatcatlurecrontfaomrinnaunttrsi.tional components and ! (c) 4MdateTribitun from an automatic systen. Samples of the whseapatevecyrifiameerdtealmasin,carloyobzriegodalnoobgpyihcoHasHlpIhacftooenrst,etnotta,anld sdcehillsecostroeildnvaetdeeldesmoelnitdss,, hydrocarbons (4) CThoenrteamiarneantnos known contaminants in the food or water that would interfere with this study. (e) EEnnvviirroonmmeennttal controls for the aninal room will be rseetlattiovemaihnutmaiidnitya tofemp5e0%rat2u0r%e, ofand19aC12to-ho2u3rC, a Tight/12-hour dark cycle. (f) AAtcclTiemaasttio7ndays (8) SBaadeseleqeducattioeonnhneuoafmlbteThrestoafnAdneixbmtoardlayswaeniigmhaltsawciclolrdibnegputorcHhHaIsedSOPsso. thAant nanoimaanlismalwililn boebvpiloauscleyd piontoor hsetauldtyhgriosuppslacuesdingonatest. The ) dsatyrsatioffiesdtudbyodyinwietiigahttionr.andomization program within nine 000118 000038 Page 32 of 38 TPa084. EXT Y Page 6 (9) Justification for Species Selection Htihsetoarniicmaalllyo,f cthheoicNeewbeZceaaluasned oWfhitthee alalrbgienoamroaubnbitt ofhas been background information on this species. B. Dose Aduinistration (1) Test Groups Group 1 (Control) 23 ((LMoewd)ium) 4s ((HHiigghh)) Test/Control Material Sterile water T-T6-2602646 T-T6-6224466 Dose Level (ma/kq) 0 105 150000 Nuomfb Aniemalrs Males Females 4 4 4 44 44 44 a The dose volume will be 0.5 mL/kg of body weight. b TTwhoe arenmiamianlisn/gsexa/ndiomsaelsle(vtewlo wainlilmables/sseaxc/rdiofsiecedlevoenl)Dawyil1l5. be sacrificed on Day 28. , C. Dosing Procedures (1) Dosing Route Intravenous injection into the marginal ear vein of the right ear over approximately 30 to 60 seconds. (2) Reason for Dosing Route sIynsttreamviecnouesxpoisnujreec.tion is an acceptable route to assess (3) Dosing Duration Single dose (4) Dose Preparation The day of treatment will be designated as Day 1. The Group 1 animals will be treated with sterile water at a dose volume of 0.5 mL/kg. The test material will be dciolnucteendtrawtiitohn stfeorrileeacwhatdeorse tolevaeclhieivneGraousppseci2f-5i.c Individual doses will be calculated based on the animal's body weight taken just before test material aadtmirnoiosmtrtaetmipoenr.aturTehe upntrielparaeddmintiessttramtiixotnu.res will be stored 000119 00003 Page 33 of 38 i 0. Observation of Animals PPa8g0e847.EXT (1) ClThieniacnailmOalbssewrivlaltiboensobserved for toxicity before test or control mactleirniicaall asdimginnsistorf ation, a(tDayapp1)r,oxiamndatedlayily0.5t,he2r.e0a,fte4.r0fhoorurcslinpiocsatl-insjiegcnstioannd twice dsaaiclryifi(ca.em. inatnerdvapl.n.()Dayfor1S moorrtaDlaiyty28).untilObstehervastcihoendsulemday be extended when directed by the Study Director. (2) FBoordyrWaenidogmhitzsation, before test or control material i(nDjayect1i5onor(DDaayy 218);), atandtheatscuhnesdcuhleeddulesdacrdiefatihceanidnterval sacrifices (when survival exceeds 1 day) (3) sample Collections (a) 4F-r,eq8u-e,ncy12-, 24-, and 48-hours post-injection, on Dianytserv8a,l 15,22, and (Day 15 or at the scheduled Day 28) sacrifice (b) MBeltohododsoafnplCeoslle(catppiroonx/iNmuamtbeelryof4 mAln)imawlisll be collected aftron4-,the8-,mar1g2i-n,al24-e,ar anvdein48-(hloeufrts eapro)sto-finjalelctiaonni,malasnd DonaysDay158."andAd2d2itfioornatlhesaamnpilmeaslswislclhedbuelecdollfeocrted on sacrifice on Day 28. Adpopcruonxeinntaetdeliyn 2t0hemLraowfdabtlao)odwi(lalctubael obvtoaliumneedtofrobem the psoasctreirfiiocre v(eDnaay c15avaoraDtaythe28)t.ineApopfrotxheimastcehleydul2e0dnl of btlhoeodstwuidlyl, baelsoc,olliefctpeodssifbrloen.moribund animals during tThheenscaemnptlreisfuwgieldl,beandstotrheedseatparraotoem tseemrpuenraatnudrecealnldular tfreamcpteiroantsurestoorfed-20inCa+1f0reCe.zerThseet steopamraaitnetdaisneruam and ctehelluSlpaornsofrracatfitoenrsewxiplelrimbeentsaelnttefrrmoizneantioonn.dryThiece to dSpiosnpsoosritiiosnroefspotnhesibslaemplfeosr the retention and 000120 00030 Page 34 of 38 ) P8084. EXT Page 8 Samples will be shipped tos JManEesE0. BJPohCnson B9l3d5g.Bus2h-36A-v0e9nue St. Paul, MN. 55106 JtaenleesphoD.neJoathns(o6n12)or7h7i8s-52a9l4terpnraitoer wtiollthebesnhoitpimfeintedofby the samples. E. Termination (1) AnUynscahneidnualleddySiancgridfuirceisngantdheDesattuhdsy or sacrificed ina gmroorsisbunndeccroopnsdyitieoxnamwiinlaltiobne sanudbjealcltedabtnooramnaliatbibersevwiialtledbe arenceosrtdheedt.izedAnwiimtahlssoindiaunmopreinbtuobnadrbciotnadliti(ovniawiilnljebcetion in tehxesanmgaurigniantaeld.earTivsesiune)s,, balseddevsiacritbheedveinnasceacvtai,onand 7a.nEi.mal(3)dyiSnagmpdleuriCnoglletchteiosnt,udwyilolr bseacrcioflilcecetdedinfarommorainbyund condition. "After necropsy, the animals will be discarded. (2) OSnchDeadyule1d5, Sathceriffiicresst two animals/sex Tevel (based on the group assignment rasasnidgonmeidzattioone)achwildlose bien athneesmtahregtiinzaeld weairthvesiond)i,umblpeedntovbiaarbtihtealven(aviacavian,jeacntdion Weixlslangbueinaanteesdt.hetiThzeedrweimtahinisnogdiutmwo peanntionbaalrsb/isteaxl/do(sveia level cianvjae,ctiaonnd einxsathnegumianragtiendaloneaDrayve2i8.n),'Abnleadbbrveiaviattheedvegnraass n(eascrdopessycriebxeadminiantisoenctiwoilnl 7.no.t b(e3)doSnaem,plehowCeovlelre,ctitoins)suewsill be collected. (3) TShaemplwheolCeollievcetri,onbile, and both kidneys fron each animal Wbiillle),beancdolliemcmteeddi,atweeliyghpeldace(dvolinunea fonrleyezedretesretminteodmafionrtain athetemanpienraaltsurewilolf b-e20diCsc+a10rdCe.d After sample collection, oTnhedrsyampilceesto(litvheer,Spobnisleo,r aafndterkidenxepyesr)imewnitllal betersmeinntatfiroonz.en J The will samples and be shipped ttoheitrhecoprerresosnponldiisntgedweiinghStesctioornvolumes a7.n0d.(d3i)s.p(ob)s.itionTheofSpthoensosrampilsesr.esponsible for the retention 000121 0041 Page 35 of 38 P8084. EXT Page 9 ) F. Statistical Analyses No statistical analyses are required. 8. Regort A final report including those items listed below will be submitted. Procedures Description of the test and control materials Description of the test system . Dates of experimental initiation and termination Description of any toxic effects Director) Gross pathology findings (if applicable) Gross pathology report (if applicable and requested by the Study Individual animal tissue weights and bile volumes 9. Location of Raw Data, Records,and Final Report Ofraicgiilniatlatedataau,ditoirngcoptihees sttuhdeyreodfu,rinwigllitsbepraovgarielsasbleandat bHeWfIoreto acceptance of the final report. When the final report is completed, all original paper data, including those item listed below will be retained in the archives of HWI according to HWI SOP. | PDroosteocporlepaarnadtipornotorceoclordasmendments In-`Bloidfyeweriegcohrtdss SamDOpoblsseeervcaaodtlmilioenncistsitornatiroencords Shipping records SPtautdhyolocgoyrreRsepcoornddsence Final report (original signed copy) The following supporting records will be retained at HNI but will not be archived with the study data. Animal receipt/acclimation records Water analysis records Animal room temperature and humidity records Refrigerator and freezer temperature records Instrument calibration and maintenance records J) 000122 CeO Page 36 of 38 PPa8g0e841.0EXT PROTOCOL APPROVAL Per 2 Luibff SHJohn L. Butenhoff, PhD Sponsor's Representative . SStteuvdeynDiM.recGtloazra , AHcauztleetoTnoxiWcioscloongsyin, Inc. Teprasentative Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-159.protdsk2) Y-995 Date 330s Date we 3-30-95 ; 0 00123 00004: - @HazLeTon Do IEISSNNRS0N Page 37 of 38 3 CORNINGCorsa PROTOCOL TP8084. EXT Single-Dose IntrTa-v6e2n4o6usinPhRaarbmbaictoskinetic Study of HHT 6329-159 Sponsor 3MMeTdoixciacloloDgeyparStenrevnitce 3PM.0.CenBtoexr,332B2l0dg. 20-26-02 St.Paul, MN 55133-3220 Contractor 3H3a0z1letKoinnsmWainscoBnosuilne,varIdnc. Madison, WI 53704 Sponsor's Representative John -_-- L. Butenhoff, Pho Study Director Steven M. Glaza ) Amendment No. 1 This anendnent modifies the following portions of the protocol: Effective Apri 4, 1995 1. cTPhoaengecetn6ets.rt7atmiaotEnexrpienareliemdemenditxattluoreDuetsifiloairnz:eGroBa.updoDso5esecvoouAlldudmmienniostotfrba0et.i5pornme:ip/a(kr1ge.)d TatestthGeroups. Modify footnote "a" in this section with the following underlined addition: a T1h-e4 daonsde0.v7o5lunmel/kwaillforbeGr0o.5upnL5./kg of body weight for Groups ) 000124 000044 EPXhPoRnEeSSIM0A3I2L01DEaLeIsVERY 3301 KINSMax 8LvO MAGIFSaONx. sw0is2s1s92i2es7 Page 38 of 38 ) Amendment No. 1 Page's HWI 6329-159 Effective April 5, 1995 2 APnaignea were l6, s a No7.. cri f EFx5p4e0r7i6me(nGtraolu iced on Days p De5simgalne;) 1 and 2, B. andDosAneimAadlminNio.strFa5t4i1o0n6; (GroupTes4tmaGlroeu)ps. respectively, due to injuries (apparent broken backs). Add the following paragraph to this section: Due to the sacrifice on Day 1 of one Group 5 male (Animal No. F54076) and on Day 2 of one Group 4 male (Animal No. F54106) because of injuries (apparent broken backs), replacement animals will be treated at the same dose levels in the same manner as for the initial animals in the study. The observations (clinical observations, body weights and sample collections) and the termination of the animals (unscheduled sacrifices and deaths, scheduled sacrifices, and sample collection) will be conducted in the same manner as for the other animals in the study terminating on Day 15. i PROTOCOL AMENDMENT APPROVAL Plm 2 BiBaubly SJophonnsLo.rsButReenphroafsfe,htaPrhsDve 3M Toxicology Service Medical Department caer Date Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. S4-139F Date y HRQeuapalrTeeitstaeynntAasWistsiicrjoaennscien,Unintc (6329-159.Am1 .dsk3) 4.18 9 Date 000125 0000% 9.1.2 Analytical protocol AMDT-042095.1 000126 000046 3M Environmental Laboratory Protocol - Analytical Study Single-dose Intravenous Pharmacokinetic Study ofT-6246 in Rabbits In-Vivo Study Reference Number: HWI#6329-159 Study Number: AMDT-042095.1 Test Substance: FC-95 (T-6246) Name and Address of Sponsor: 3M SCD Division 3St6.7PGaurlo,vMe NStre5e5t106 Name and Address of Testing Facility: 3M Environmental Technology and Services 935 Bush Avenue St. Paul, MN 55106 Proposed Initiation Date: July 25, 1995 Proposed Completion Date: August 25, 1995 Method Numbers and Revisions: AMDT-M-1-0, Thermal Extraction of Fluoride by Means ofa Modified Dohrmann DX2000 Organic Halide Analyzer-Liver AMDT-M-2-0, Fluoride Measurement by Means of an Orion EA940 Expandable Ton Analyzer AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver AMDT-M-8-0, Analysis of Fluoride Using the Skalar Segmented Flow Analyzer with lon Selective Electrode AMDT-M-14-0, Thermal Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum Author: James D. Johnson PRL fames D/Jhnson Study Difector ffts Date GE LE dat) fester John Butenhoff, PhD Date Sponsor 000127 000047 1 10 PURPOSE This study is designed to provide information as to whether the perfluorooctanesulfonate anion does go to the rabbit liver and other tissues when the material wisitahdmtiinmiestaefrteerd dionsaen iinnstrearvuemnoaunsddloisvee,r.and to ascertain the change in concentration 2. EST MATERIALS 2.1 Test, Control, and Reference Substances and Matrices 2e.q1u.i1valAennatl.ytical Reference Substance: FC-95, lot 161 or 171. They are 2.1.2 Analytical Reference Matrix: Bovine liver and bovine serum 22..11..43 AAnnaallyyttiiccaall CCoonnttrrooll SMuabtsrtiaxn:ceB:ovNinoeneliver and bovine serum 2.2 Source of Materials: 3M ICP/PCP Division (2.1.1), grocery store (2.1.2, 2.1.4 liver), Sigma Chemical Company (2.1.2, 2.1.4 serum) 2.3 Number of Test and Control Samples: Liver and serum from 32 test animals and 8 control animals. Two animals were replaced at 24 hours (animal F54076 was replaced with animal F54112, and animal F54106 was replaced with animal F54099). Other biological tissues (kidney, bile, cellular fraction) will be available for analysis if deemed appropriate by the Study Director. 2.4 Identification of Test and Control Samples: The samples are identified using tnhuembHeWr,I palnuismtahle itdiesnstuiefiicdaetnitointynaunmdbedraywhiidecnhtictoyns(issetrsumo).fa letter and five digit 2.5 Purity and Strength ofReference Substance: To be determined by Sponsor. 2.6 Stability of Reference Substance: To be determined by Sponsor. 2.7 Storage Conditions for Test Materials: Room temperature (2.1.1), -20 10C (2.1.2, 2.1.4). Test and Control samples will be received according to AMDT-S-10-0. 000128 UC0as 2 2G.L8PDiRsepgousliattiioonnofforSptehecitmiemnesp:erBiioodlroegqiuciarletdisfsouresstaundidesflluoidnsgewriltlhabne 2r8etdaaiynse.d per 2.9 Safety attire. Use Precautions: caution when Refer to handling akpnpirvoepsrifaotrecuMtStiDnSg.theWesaamrplaepsp.ropriate laboratory 3.0 EXPERIMENTAL - Overview The tissues from animals dosed as described (HWI#6329-159), are available for analysis for fluorine compounds. At the discretionof the Study Director, a series of a(nSaeleytMiectahlotdess-ts-nceaxntbSeecpteirofno)rmisedt.he Tahpperospcrrieaetneinagnafloyrsifslutooripdreesiennltivdeerfivniiaticvoemdbautsatifoorn falpuporroixniemiantithoen ltioverra.bbFiotrlitvheersefoarnmalaytsreisx,sbpoivkiesneanlidverrecaopvpeerayrsdettoebremianagtoioodns. Since pacecrufrlautoeroeoscttiamnaetseuolffotnhaetecoanncieonntriastniootn boifottrheancsofomrpmoeudn,df.luorine content will be an 4.0 EXPERIMENTAL - Methods 4.1 Liver and Serum screening methods: (attached) ` 4.1.1 AMDT-M-1-0, Thermal Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver 4E.x1p.a2ndAaMbDlTe-fMo-n2A-n0al,yFzleuroride Measurement by Means of an Orion EA940 4.1.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver 4.1.4 AMDT-M-8-0, Analysis of Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 4.1.5 AMDT-M-14-0, Thermal Extraction of Fluoride by Meansof a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum 5.0 DATA ANALYSIS 5.1 Data Reporting: fluoride per tissue or Data fluid, will or as be reported FC-95 per taisssauecoonrcfelnutirda.tiSotnat(iswteiicgshuts/ewde,igahttt)heof discretionofthe Study Director, may include regression analysis of serum concentrations with time and averages and standard deviations of concentrations for 000129 000019 3 mdiafyferbeentapdpolsieedgrtooupdse.terIfminneceesstsaatriyst,icsailmdpilfefesrteanticset.icaHlaltfe-sltisfseuofchdiassaSptpuedaernatnscet tferstom serum will be determined from the linear regression of the averages of the concentrations at different time points for the post distribution part ofthe serum concentration versus time curve. 6R 0MA AINWTED NANA CEOTF AANDRECORDS 6.1 Raw approved Data final and Records: Raw report, and electronic data, data approved protocol, will be maintained appropriate specimens, in the AMDT Archives. 7.0 REFERENCES ~~~ 7.1 AMDT-S-10-0, Sample Tracking System 8.0 ATTACHMENTS 8.1 AMDT-M-1-0, Thermal Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver 8.2 AMDT-M-2-0, Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer 8.3 AMDT-M4-0, Extraction of Fluorochemicals from Rabbit Liver 8.4 AMDT-M-8-0, AnalysisofFluoride Using the Skalar Segmented Flow Analyzer with Ton Selective Electrode 8.5 AMDT-M-14-0, Thermal Extraction of Fluoride by Meansof a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum 000130 CO0S0 4 3M Environmental Laboratory Method Thermal ExtractionofFOrlguaonriicdeHablyidMeeaAnnaslyozfaerM-oLdiivfeired Dohrmann DX2000 Method Identification Number: AMDT-M-1 Revision Number: 0 Adoption Date: /o- -15~ Revision Date: None 0 -_-- Author: Rich Youngblom Approved by: - fern Zoup Leads? / { Lip mis 2 Quality Assurance /fof3/3s Dale L0oo-s4-7 Date Software: MS Word 5.12 Affected Documents: AMDTE-xMp-a2ndaFbllueorliodneAMneaalsyuzreerment by Meansofan Orion EA940 AMDT-EP-3 Organic Routine Maintenance Halide Analyzer ofa Modified Dohrmann DX2000 000131 1 Ge00%1 1.0 SCOPE, APPLICABLE COMPOUNDS, AND MATRICES f1l.1uoSricdoepef:roTmhivsarmieotushomdatirsicfeosr.thTehoepefrlautoiriodneoi fatyDpiocharllmyacnonllDeXct2e0d0in0TwIhSeAnBitsiosluutsieodntfoorexatnraalcytsis with an fon selective electrode. 11..32 MAaptprliicceasb:leBiCoolomgpicoaulntdisss:ueFs,lupoarrtoicchuelmarilcyallisveor.r other fluorinated compounds. 2.0 KEYWORDS 2D.X12F0l0u0or,idfel,uofrloucorhienmei,ceaxltsr.action, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, 3.0 PRECAUTIONS 3.1 3.2 Glassware Glassware and exhaust gases can is fragile, broken glass be extremely hot, may cause injuries. 33..34 SPorlesvseunrtizbeadsegdasseasm,pplreospemracyomflparshe,ssmeadygnaesehdantdolailnlgowprtachteimcetsorderqyuidreodw.n before starting run. 3.5 Potential equipment. biohazards due to the biological matrices. Use appropriate personal protective 4.0 ND MATER] 4.1 4.2 Compressed Compressed Oxygen, Helium, HHiygdhroPcuarirtbyoGnrfardeee,, regulatteod regulated to 30 45 PSL. PSL. 4.3 4.4 Quartz Quartz glass glass sample boat combustion with tube, TeflonTM Reliance tubing, Dohrmann Glass G-9405-012 890-097 or equivalent. or equivalent. 4.5 4.6 SOraimopnle94c0ol9l9e9ctTiootnavliallosn,icHDSPtrEe.ngth Adjustment Buffer (TISAB IT) or equivalent. 4.7 Milli-QTM water 4.8 Polystyrene pipettes. 44..910AcHtaimvialtteodnCShyarriconagle,oEr.eMqueirvcalken2t0.05 or equivalent, 4.11 Miscellaneous laboratory glassware 5.0 EQUIPMENT 5.1 5.2 IRoBsMemcoomupnattiDbolher3m8a6nnorD4X8260c0o0mpOurtgearn.ic Halide Analyzer, modified for fluoride extraction. 5.3 5.4 EDxXc2e0l 0S0presaofdtswhaereet,, version version 1.00, modified 5.0 or greater for fluoride extraction. 6.0 INTERFERENCES 6m.a1ySvaamrpylferosimzemaitsrliixmittoedmattoriaxp.proximately 150 mg, depending on sample moisture content. This 000132 2 000035 7.0 SAMPLE HANDLING 7s.a1mpSlae.mplUesseafroerncoetpstoorbeprhoabnedlteodtrwaintshfebratriesshuaensd.s. Fluoride may leach from the skin to the c7l.2eaSnasmcpallepeslofanlidvceurtatriengcbuotarfdr.omTfhreozceunttliinvgerbaoanrddpalnacdesdcailnpaeltasrheoduladndbelacbleelaendewdewiigthh bwoaatt,er,Use a methanol, or methanol-water solution after each liver is cut. 8.0 CALIBRATION AND STANDARDIZATION 8.1 PreparationofCalibration Standards 8R.e1f.e1rTthoeprsottaoncdoalrfdosrtrheqautirpreodjefcotr.cach project will need to be appropriate for that individual project. 88..11..32 TFyoprircaablbliyt 5li0v-e5r0s0tupdipesm,FuCs-eb9e5eifnlmiveetrhaansotlhestmaatnrdiaxr.dsCaurte auspedi.ece of frozenbeefliver (100 150 mg) and weigh it in a labeled and tared weigh boat. 8.2 Calibration - Overview `cTuhrevenoirsmraelquciarleidbrtahteiponroicsetdhuerfeluliosrtieddebceulrovwe (i AuMsDedT.-M-2). However,ifan optional spiked liver 8a.n2d.1coAmbcuasltiibrnagtitohnecmuursvienfgorthtehesaDmXe2m0e0t0hoisdsgeannedramtaetdribxy tsyppiekaisngthseamspalmepslewsittho kbenotewsntesd.tandards 8.2.2 Typically, spiked. three replicatesof cach standard and five concentrationsof standards will be 8R.e2c.o3veSrteadndFar(dugc)uornvethwielyl-abxeisp.loGtteenderaastMeaassreSgpreiskseidonFc(uurgv)e on the x-axis and calculate and the Standard equation Mass for the line: a8.n2d.4thMears2svaSlpuie.ked F (ug) = (Amount spiked in mL) x ( Conc. of standard in ppm) x (0.6004) 8.2.5 Standard"MFaCs-s95Reicso6v0e.r0e4d%FF(tuhge)re=fo(rTeI0S.A6B00v4oilsutmhee finacmtoLr)usxe(dOtroiocnonrveeardtinFgC-i9n5pptom)F. 8.3 Calibration - Procedure 8.3.1 Start Up 8b.e3.u1s.e1diRfutnhe2porrevmioorues sCalmepalneCsyccolnetsaiwnheednhistgahrtcionngceinntsrtartuimoennstoefafclhuordaiyd.e. More clean cycles may 8.32 Blanks 8.3.2.1 8.3.2.2 PFroerpraarbebistasmtpuldieesu,siunsge tbheeesf alimveermaesthtohedsmaatnridx.typPeroefpamraetratixleaasstt3hestaemsptlseasmopfleb.eefliver 8(31.020.3 1P5u0tmsga)mpfloer bilnanDkos.hrmann boat. Combust each sample as described in section 9.0 and analyze sample according to method AMDT-M-2 for the ion selective electrode analysis 000133 3 00005. 8b.e3f.o2r.e4prFoocreredaibnbgi swtiutdhietsh,e tchaleibmreatteiorn.reaBdiunmg fsoarmpalbelsanukntislamthpilselismhiotuilsdrebaec0h.e0d3, oprpumntoirl linowteher judgement hours). of the operator the reading is stable with respect to historical readings (previous 43 83.2.5 before For non-rabbit proceeding with stthuediceasl,ibtrhaetiboln.ank readings should reach a predetermined ion concentration 8.3.2.6 It may combustion. be necessary to mix approximately 50 mgof charcoal with the sample to aid 83.3 Standard Curve 8l.a3b.e3l.e1d wWeeiigghhbooautts.atlFeoarstra1b5bimtatsrtiudxiessa,mpwleeisgh(51s0t0a-n1d5a0rdmsgwbiethef3lrievpelricsaatmepsleeasc.h)Reincotrardedweaingdhts in s8t3u3d.y2dataP.lacSetowreeitghheemdabtereifxlsiavmeprlseasmopnledriyn iDcoehorrmiacnenpascakmspltoekbeoaetp.them frozen until used. q8u3a.nt3i3tySotfartthweistthantdhaerldoowensotrsitnantdhaermdatcroinxc.entFroatriroanb.bitUssitundgieas,Haumseil4tounLsoyfrisntgaen,deajredctanadfeijxeecdt it on or in the beef liver. 8.3.3.4 may be Atleast 3 replicates should used at the discretionofthe baenaluyssetd. for the lowest standard concentration; more replicates 8.3.3.5 83.3.6 RCuonmbaullst15thsetasnadmarpdlse. Iasfdoensecrriebpeldicianteseisctsiiognni9f.i3caanntldyadniaflfyerzeentacfcroormditnhge toothAeMrDtTwo-M-2. roelpdlirceaptleisc,atreunanadnotthahterthseamnpelwerfeoprlitchaatteswtialnldabred.usIenddtiocactaelciunldaatteatthheartetghreesnseiwonrceuprlviec.ate replaces the 8le.a3s.t3.07.95W,hceonnsaullltstwaintdhasrudpserhvaivseorb.een run, calculate the 12. 12 must be at least 0.95. If itis not at 83.3.8 A changed. new New standard standard curve curve mshaoyuladlsboebreunruwnhaet nthtehedicsocrmebtuisotnoifotnhteubaenaolryssta. mple matrix is 8.4 Storage Conditions for Standards 8T.y4p.i1caSltloyr,asgteanrdeaqrudisreamreensttsorfeodr sattarnodoarmdsteamrpeedraetpureenidneponlnatsttihcesicnrdeiwvitdoupalbostttalensd.ards used. 8.4.2 New FC-95 standards should be prepared at least once a month. 9.0 PROCEDURES 9.1 Typical Operating Conditions: 9.1.1 9.1.2 Combustion Oxygen and Htuebleituemmfpleroawt=ur5e0=cc9/5m0inCt.e. 9.1.3 9.1.4 VBaapkoeritziamteio=n3/D0r0ysiencgontdism.e = 240 seconds 9.2 Start Up Procedure: 9.2.1 9.2.2 If the Open ptrhoegSraYmSiTsEnoMt SstEarTtUedP, swtiarntdtohwe.EOX program on the PC. 99..22..34 PCultostehethfeumSaYceSTmoEdMulSeEaTnUdPthweicnedllowi.n the READY mode. 000134 4 00005 9:2.5 found When in the tCheEoLvLenChHaEs CreKacmheendu.the READY temperature, run the CLEAN BOAT program 9.2.6 See AMDT-EP-3 for details of the Dohrmann sofware. m99.3i3.x1SaapOmpppreolnxeitmEhaxetteSrlaAycMt5iP0oLnmEgPorHofcAcehTdaCurrcHeo:aalndwiptlhactehethseamspalmeplteo aiindtchoemBbuOsAtTio.n.It Imfatyhisbeisndeocnees,sacrhyartcooal 9s.ho3u2ldCallossoebeSAmiMxPedLEin HwhAiTleCHes.tablishing the baseline and when generating the standard curve. l9.a3b.e3ledAdsadmpaplperocporllieactteiovnolviuaml.e of TISAB Typically 0s.o6lumtiLontoor151:m1 LTIaSrAeBu:seMdi.llFio-rQrTMabwbaittesrtumdiixest,urueseto1a.0 or 92..034mLPloafce1:t1heTIvSiaAlBs:oMitlhalti-thQePtTMipwoatfetrhemiCxOtuMreB.USTION TUBE is in the TISAB at least 0.25 9i.n3c.h5es.RuGnastehse EreOlXea-sSedOdLuIrDinSgppryoroglryasmisfmouusntd binubtbhleeRtUhrNoumgehntuh.e TISAB. 9.3.6 93.7 IWfhuendniltuhteeEdOTIXSpArBogwraasmuissefdintioshceodl,lercetmtohveesatmhpelceo,llaedcdtiaonn evqiaulalfrvoomluthmeecoofmMbuisltli-oQnTMtuwbae.ter 90.3t.h8eRTiInsSeAtBhetoenmdakoef t1h:e1cToImSbAuBs:tMiiolnltiu-bQeTMw.ith Milli-QTM water and wipe with a KIMWIPE to w9ri.e3tm.ho9aveOcpoateninyontThtIeiSpspAaeBmdpralpeepmhlaaiticcnahtiaooonnrrngd vrtahecemuotuuvbmeee.adnoyutr.emIatimnaiyngbaesnhecfersosmarthyetobosatc.rapAspahrtciaclnesboefrfetmhoeved bbooatttotomawiidtihnathsepaCtluelaanorCyoctlhee.r similar device. A drop of Milli-QTM water may be added to the 9.3.10 Close the hatch. 9.3.11 9.3.12 SRaumnplteheisCrLeEadAyNfoBr OanAalTyspirsobgyraimo.n selective electrode (AMDT-M-2). 9.4 Sample Caleulations 9.4.1 94.2 Use the Sample sMtaasndsaRrdeccouvreveretdo Fcal(cuugl)at=e(tTheISsaAmBplvoelvianlume,L) x(Qrionreading inppm -intercept) (Slope) 10.0 VALIDATION 1100..11.1QuDaailliytyStCaornttrUoplCheck Samples: Once the standard curve is established, each day of caonnaclyesnitsraitsisotnarstpeidkebdy saanmapllyzeisngusQedCtsoagmepnleersa.teTthhee sQtCandsaarmdplcuersvea.re Etoacbhe ctohnecsenatmreataisonthmeulsotwebes.t rdeopnleicaintetriispnloictatneecuenslseasrsy.the first two replicates are within 20% of the standard curve, then a third F1l0u2oriPdreeNciostieobnooaknsd 2A,c3,cuarnadc5y.: PSreeceimsieotnhaondddaecvceulroapcmyevnatriaensalwyhsiesnaanndalsyazmipnlgesaanmapllyessiosfidnifferent matrices and different reference compounds. 10.3 Other Validation Parameters: NA 000135 5 0000: 11.0 DATA ANALYSIS 111 Calculations ILL 11.1.2 TFoorctahlecusltaatnedtahred fcluurovrei,deusceonrtergarcetsisoinoninantahelyssaimspilne,Exscceel,mevterhsoidonA5M.D0To-rMg-re2at.er. 11.2 Analyzing the Data a1r1e.2w.i1thi2nm2u0s%t obfe eataclheaostth0e.r9a5nodrtghreeaotuetrl.ie"rOuistlgireerast"ermathyanb2e0e0x%cloufdethdeiaftvewroaogfetohfeththorseeetrweoploircates nloestsedthiannt5he0F%ionfatlhReepaovretraagleoonfgtwhiotshethtewor.easAonnyitswuacsh ocuotnlsiierdsersehdoualndoubtelipeori.nted out in the data and. 2.0 A ENT: None 13.0 REFERENCES 13.1 349, Rosemount revision B, Dohrmann December DX2000 1993) Organic Halide Analyzer Operator's Manual (Manual 915- 13.2 AMDT-M-2 Analyzer Fluoride Measurement by Means of an Orion EA940 Expandable fon 13.3 AMDT-EP-3 Routine Maintenance ofa Modified Dohrmann DX2000 Organic Halide Analyzer 14.0_REVISIONS Revision Number ReasonforChange Revision Date 000136 & C0003_ 3M Environmental Laboratory Method Fluoride Measurement by Means of an Orion EA940 Expandable Ton Analyzer Method Identification Number: AMDT-M-2 Revision Number: 0 EE Author: Rich Youngblom Approved By: Loner Gro] Leader Adoption Date: /0- 4-55" Revision Date: None , VAY AS Date Le LAL Quality Assurance fot Date Software: MS Word S.1a Affected Documents: AMDT-M-1 Thermal Extraction of Fluoride by Meansof a Modified Dohrmann DX2000 Organic Halide Analyzer 000137 1 00005, APPLICABI D 1. Ton SAnCaOlyPzEe:r. This method is for the calibration and operationof an Orion EA940 Expandable : 1.2 APPLICABLE COMPOUNDS: Fluoride. P1.H3OAfP6.P0L.ICABLE MATRICES: Liquid samples in an appropriate buffer solution. Preferred 2.0 KEYWORDS 2.1 Fluoride, fluorine, ion selective electrode 3.0 PRECAUTIONS 3.1 No hazards identified with this method. 4.0 SUPPLIES AND MATERIALS 4.1 4.2 Orion Orion 940999 Total Model 900001 TeolneicctSrtodreenfgitlhlinAgdsjoulsuttmieonnt(ABguCffle)roIrT (TISABII) equivalent. or equivalent. 44..43 OMriilolni-9Q4TM09w0a7te1r0o0rpeqpumivafllueonrti.de standard or equivalent. 4.5 Magnetic stir bars. 4.6 Lab tissues. 44..87 SPlaamsptilce1c0o0llmecLtiovnolvuiamles.tric flasks. 44..910PoMliysscteylrleanneeopuipsetltaebso.ratory glassware. 5.0 EQUIPMENT 5.1 5:2 Orion Orion Model Model EA940 960900 ESxolpiadndStaabtleeCloomnbiAnnaatlyizoenrFolruoerqiudievaelleenct,trode or equivalent. 5.3 5.4 IMaBgMnectoimcpaSttiirbPlleat3e8.6 or 436 computer (only neededif using Orion 3E software). 55..56 OMirciroonsRofSt23Ex2ceinlte5r.f0a(ceonclayblneee(doenldiyfneuesdiengdiOfruisoinngJEOrsiofotnwa3rEe)s.oftware). 6.0 INTERFERENCES Q6.1MwItaitserrewciolmlmgeenndereadlltyhabtrtihneg psHambpeleattoorpnHeoafr66..00.. A 1:1 mixtureof TISAB and sample/Millia6t.2roSoammptleempteermapteurraetuarsethmeasytaenfdfeacrtdsflwueorriedewhmeeanstuhreemmeetnetr. wIatsiscarleicbroamtemde.nded that the sample be 6.3 The rate the samples are stirred at should be consistent with the rate the standards were stirred. 000138 2 000! 0: u6:n4deArirelbeuctbrboldees. trapped under electrode can give erroneous readings. Make sure no air is trapped 2.0 SAMPLE HANDLING 7.1 No special handling necessary. 8.0 CALIBRATION AND STANDARDIZATION 88..11.1PrMeepaasruarteio5n0 mofLCaolfiTbrIaStAiBonIStinatnoda5r1d0s0 mL plastic volumetric flasks. 8.1.2 8.1.3 PLiapbeetlteth0e.0f5l,as0k.s1,250.05.,051,.0,0.a1n,d0.15.,5m1.L0, aonfd110.05 ppppm fFl-u,oarildoengstwaintdhartdheindtaotethaenadppyroouprriinaitteilalys. labeled flasks 8.1.4 8.1.5 SAhdadkeaptphreoxfliamsaktsetloym3i0xmtLhe of Milli-Q solutions. waterto each flask. 8.1.6 Eliminateair the flasks over the abiurbbbulbebslefs.rom the flasks by tipping the flasks on their sides and rolling the air in 8.1.7 8.1.8 Bring Invert the volume in and shake the the flasks flasks for up the to the 100 mL final mixing, mark with Milli-QTM water. 8.1.9 Record standards in Standards Log Book. 8.2 Calibration 8.2.1 8.2.2 IIfnvneerctespsraorbye, troewmeotvetotpapseealf.rom electrode filling hole. 88..22..43 FEijlelctthae efleewctdrroodpeswoifthfiflillilnigngsoslouluttiioonn.from bottom of electrode to wet lower seal. c8o.r2r.e5cTthioen,meutseirnganstdatnhdearFd-sewlietchtrcoodnecaernetrtaytpiiocnalsolyf cal0i.b05r,a0t.e1d,b0y.5d,ir1e.c0,t maenads1u.r5epmepnmtFw-,itfhonlloobwlianngk 8t.h2e.m6aRneufcaocrtdurteher'ssloipnestirnuctthieonasp.propriate log book. 8.2.7 Clean tissues the electrode by rinsing with Milli-QTM water and wiping the sides down with lab 88.33.1SCtaolriabgreatiCoonndsittanidoanrsdsfoarreStstaonrdeadradtsroom temperature. 9.0 PROCEDURES 99..11.1CaTlhiebrsaatmipolneatnodbeMemaesausrueremdennte,edSsttaonbdearmdixmeedthwoitdh:TISAB using the proportions 9r.e1c.2omPmlaecnedaedstibrybatrheinTIthSeAsBammpalneufaancdtuprlearc.e the sample on the stir plate. e9.l1e.c3trAodleloiswitnhseerstaedm,plmeaktoemsiuxreftohrearefaerwesneocoainrdsbubbebfloersetirnaspepretdinugntdheeretlehceterloedcet.rodWe.hen the 9.1.4 same The rate sample should be as the calibration the same standards. temperature as the calibration standards and stirred at the 9.1.5 When the readings have stabilized, record the reading in the appropriate log book. 000139 3 00007 9.2 Calibration And Measurement, Using Orion 3E Software: 9.2.1 Calibration: 9.2.11 Follow steps 8.2.1 10. 8.2.4. 9.2.1.2 Press Function Key #8 (F8). c9o.n2.c1e.n3trTahteiocnoomfptuhteesrtsancdraeredns,wialnldaswkheytohuertoocronnoftiarmbltahneknis tuo bmeoifnsbctlauneddeadrrdinstthoe bcaeliubsreadt,ion. 9M.a2.k1e.4aPnlyacneectehsesaerleyccthroadnegeisn tthoethfiersitnsftoarnmdaatridononprtehseesnttredplaantde aclnidckcloinckCoOnNCTOINNTUIEN.UE. s9t.a2b.i1l.i5zeOdb,sperrevsestAheCrCeEadPiTngsRoEnADthIeNgGra.phic display on the computer. When the readings have 99..22..11..76 ARfetpeeratthsetfeipna9l.2s.t1a.n4d.aarndd,9t.h2.e1.c5 ompfourtethrewrilelmadiinspilnagystthaendsalrodsp.e of the curve, as well as the intercept and click aonndCcOorNreTlIatNioUn.E.ReTchoerdcatlhiebrsaltoipoen,dianttaericsepatu,toamnadticcoarlrlelyactioopnieidn the appropriate log book to C:\Orion\Data\Calib.txt. 9.2.2 Data Spreadsheet: 9.2.2.1 0 store Select cither data in. NEW or OPEN from the FILE menu to open a new or existing spreadsheet 9.2.2.2 Record the name of the 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 9.2.3.2 Enter the name of the sample in the appropriate place on the screen. 99..22..33..43 WClhiecnk othnetrheeaNdiEngWs hSaAveMsPtLabEilibzuetdt,ocnlick on the RECORD button and write the result in the. appropriate log book. .0_VALIDATI 10.1 Quality Control: 10.2 Precision and Accuracy 103 ppm fOltuohreirdeVuaplitdoataisoantuPraatreadmesotleurtsionAocfcofrludoirindge,to Reference 13.2, the rangeofdetection is 0.02 11.0 DATA ANALYSIS 11.1 Calculations None necessary. 11.2 Analyzing the Data None necessary. 12.0 ATTACHMENTS None 13.0 REFERENCES 000140 4 coee" 13.1 Orion Model Incorporated, 1991 EA940 Expandable lon Analyzer Instruction Manual, Orion Research 13.2 Orion Model 960900 Solid Research Incorporated, 1991 State Combination Fluoride Electrode Instruction Manual, Orion 14.0 REVISIONS Revision Number ReasonforChange Revision Dae 000141 5 0000 3M Environmental Laboratory Method Extraction of Fluorochemicals from Rabbit Livers SOP Identification Number: AMDT-M-4 Adoption Date: /o-7/-55 Revision Number: 0 Revision Date: None r ----e -------------------------- Author: Dave Christenson/Cynthia Weber Approved By: froup Leader Jo--3/-25 Dae "ue Le Fd Quality Assurance lf=re15" Date MSAfoaffsetscwtaSerpdee:cDtorMcouSsmceoWnpoytr.sd:, 6M.-05, AnaloyfRsabibist ExtractforFluorochemicals Using Electrospray 000142 00008: | 1.1 SEtchoyple:acTehtaitsemies tuhseodd tios efxotrrtahcet efxlturoarcotcihoenmiocfafllsuforroocmhtehmeiclaivlesrsfrfoormarnaablbyistislibveyrs. 1.2 Aelpepcltircoaspbrlaey Cmoamssposupencdtsro:scFolpuyo.rochemicals or other fluorinated compounds. 1.3 Matrices: Rabbit Livers. 22.0 1 FluorK ochemicalE s, rabbit lY ivers, elecW trospra~ y mO ass~ spect~ rR ometer, flD uorinatedS compounds, extraction. 3.1 Use gloves when handling the rabbit livers, they may contain pathogens. 4.0 SU4.P1PLISuEpSplAieNs DMATERIALS ~~~ 44..11..12 ESyprpienngdeo,rcfaptaybpleeoorfmdiesapsousrabilnegp1i0pe0tus L 44..11..33 GPllaosvtiecsgrinding tubes 44..11.056 LPlaabsetlisc centrifuge tubes, 15 mL 44..11..87 TNiimweorgen 44..11..910 AFnilatleyrtsi,cTailtpainpentsy:lognlassysrivnogleumfeitltreircs,p0ip.e2rsp.m. 44..11..1112 CDrisipmopsacbalpeapultaosvtiiacls3.cc syringes. 4.2 4R.e2a.g1enAtqsueous Ammonium Acetate (Aldrich), approx. 250 ppm: Prepare a 2500 pacpetmataequ10eo2u1s0s0olmuLtiovnoolfumaemtrmiocnfilausmkanacdedtialtuetbeytaodvdoilnugm2e5w0imthgMailmlmio-Qnium 4.2.2 Swaotdeir.umDiclaurtbeotnhaitses/oSloutdiiounm1B:i1c0afrobronaa2t5e0Bupfpfmers(o1l.uTt.ioBna.ker), a(nNda2,1.C0 OJgNGofHsCoOd,i)um0.bi5caMr:boWneatiegh(N2F6I.5COg,o)fisnotodaiu1mLcavroblounmaetteriPcicflaosk)and. 4.2.3 bDirliuntgetoacveotolnuimteilweistohluMtiilolni,d-iQluwtaetaecre.tonitrile 1:1 with Milli-Q water. 44..22..45 MEtehtyhlanAcoeltate 44..22..76 MIiHl,li1-HQ.w2aHte2rH - perfluorooctanesulfonic acid (Aldrich) 4.2.8 FC'95 (3M Specialty Chemical Division) 000143 000063 2 5.0EQ$5U.1IPUMloEa-NTuTmrax T25 Grinder for grinding live~ r samp~ les.~ 55..32 CVoerntueixfumgiexer 55..54 ASnhaalkyetrical Evaporator 000000 6.1 There are no known interferences at this time. 10 SA7.M1 PLThEeHrabAbiNt lDivLersIarNeGre_ce_ive_d f_ro_ze_n,_and performed. : must bekept~ froze~ n und~ l the extraction is 8S .0CT A8.L1IBA RPrAepTN arIaOtiNoD nAoNf A IDnternR al StaD ndardI s ZATION 8.1.1 pPerretplauroeroaonciinatnecrsnuallpshtoannidcaarcdidoftaopbperoaxdidmeadtetolyea1c2hpLipvme1r sHa,mp1lHe,.2H,2H8.1.2 1W0e0imghLatvloelausmtet0r.i1cgfolafsk1.HR,e1cHo,r2dH,t2hHe-apcetrufalluwoeriogohctt.anesulphonic acid into a 88..11..43 TvBoroilanugm2ei5t0wuimpthL'toMvvioolllluium-emQterwiwacittefhrl.amsCekat,lhacadundloal3t,emttLhhisesaiocsfttuthahelescstotonoccckeknststrtaaantndidaoarnrdod.fatnhdebring to standard. actusaullmphgonpiecrfalcuiodroctane- X 3mL = OIL 350 mL actual concentratipopnm, 8.2 8P.r2e.p1arPereFpCar-e95FC-A9n5iosntanSdtaardnsdaforrdtshe standard curve. 8.2.2 WReeciogrhdatphperaocxtiumaaltweeliygh1t0.0 mg of FC-95 into a100mL volumetric flask. 88..22..43 DBirliuntgeutphetsoovloutliuomnewwiitthhdidliultueteacaecteotnointirtirliele1.:10for asolution of aapsporlouxtiiomnatoeflyap1pr0o0x.ppm1.0 pDiplmu.te this solution 1:10 with dilute acetonitrile for 8.2.5 5U.s0eptphme,101.p0ppmpsmolauntdio5n0t0o make ppb. working standards with values close to 8.3 8P.r3e.p1areWeiBgehef40LigvoefrBoHvoimnoegleivneartinetofao25U0smeLfoNralSgteannedabrodrse containing 8.3.2 A20d0dm1LmsLoMiflltih-eQswoaltuetri.onGrtoinad1t5omaLhocmenotgreifnuoguestsuobleu.tPirone.pare a totalofcight s1umspLenadlisqoulouttsioofn tbhyesshoalkutiinogniitnbe1t5wmeeLncaelnitqruioftusg.e tubes. Be sure to re- 000144 000068: 3 8.3.3 Spike seven standards in of the 1 mL step 9.12 of atlhiequportoscewdiutrhe.thOe nfeoll1 omwiLngalaimqouuotntsserovfeswoarsktihen.g blank. IT wx] Working Standard ra (Approximate Conc.) "Approximate fina concentration of FC-95 in liver 8.4 Calculate the actual value of thestandards: cL.oofsntancdared xntrat(iinoppnm) = final concentration (ppm) 171 mg liver'T/ ml homogenate of FC -95 in liver *Average weight ofbovine liver in solution as determined by weighing F1 Cm-L95hiosmoregpeonratteedsasofe4qu0ivmaglelnitvseroifnF2C0-095mLpootfaMsisliluim-sQaltw.ater. The amount of 8.5 Calibration Spectrometer. 8.5.1 Extract the spiked beef liver `homogenate following 9.13 t0 9.23ofthis method. Use these standards to establish your curve on the mass 8.5.2 Altematively, a standard curve may be generated using ratios of responses coofntcheentpreartfilounoroofocthteanpseurlfflounoartoeocatnainoensualnfdontahteeinatneironna.l standard anion versus 8.6 Storage Conditions for Standards 8.6.1 cNoevwersetdapnldaasrtdisc is performed. caernetprriefupgaeretdubweisthunetialchthaenaalnyasliyss.isSotanntdhaerdmsasasresspteocrterdomienter 8.7 8S.t7o.r1ageBeeCfonlidvietrihoonmsogfeonraStetsanmdaayrbdesfrozen after preparation. 9.1 Obtain frozen liver. samples. In spent tissue, note that the liver has not been 9.2 9.3 Upasceakagdeisdsewcittihngotshcearlpteilssuaensd. cut off approximately 1 gofliver. Weigh the sample dircctly into a tared plastic grinding tube. 9.4 9-5 PRuetcaolradbethle olnivterhewveiiaglhtwiitnhtthhee ssttuuddyynnoutmebbeoro,k.weigh, rabbit ID, date and analyst initials. 000145 e00s s 99-.76 9:8 99 9.10 9.11 9.12 9.13 9.14 9.15 9.16 9.17 9.18 9.19 9-20 9:21 9.22 9-23 9-24 9.25 AGrdidnd2.t5hemsLasmpwlaet.erP.ut the minutes, until the sample igsrainhdoemropgreonbeeoiunsthseolsuatmipolnewiatnhd grind for about 2 no large chunks, TRainkseetthheegprriondbeerofafpairnttao ntdheclseaamnpilt ewiwtihthme2t.h5amnLols awfatetrereaucshinsgamappliep.etF.ollow CAaMpDTt-hEePs-a2m2p.le and vortex for 15 seconds. Pipet I mLintoa `irnefmoarimnaitnigonstaespst.h)e 1m 5 Lcentrifuge tube. Label the grinding tube. (See AMDT-M-4 centrifuge tube Worksheetfor with the identical documenting the. Spike the beef liver homogenates as described in 8.3. with the appropriate amount of FC-95 standard acetate. SApdidke1 mthLeosfamptlheessoadnidbuemecfalribvoenratheo/msoogdeinuamtbeiscawribtohna1t0eb0uufLfoefrianntedrn1 amlLsatamnmdaordn.ium Using an analytical pipet, add 5mLethyl acetate. Cap the oh and vortex 20 to 30 seconds. CPeutnutihfeumgeintfhore2s0htaoke2r5 for 20 min. minutes, until the layers are well separated. Set thepower on the centrifuge to 25. `RmeLmgorvaedu4atmeLdsgolfastshpeip1e0tp. oTrrgaannsifcerltahyeetlroabealftroesthhe15frmesLh centrifuge tube. tube with a 5 Blow the sample down on the analytical evaporator to near dryness withnitrogen, Barpipnrgoxtihmearteemlayi3n0intgo 40 minutes. sample up in 1 mL dilute acetonitrile with an analyticalpipet. TVroarntsefxer1t5hseescaonmdpsl.e to a 3 mL syringe. Attach 2 0.2 ptm nylon mesh filter, and filter the sample intoa fresh centrifuge tube study number and animal number. or a autovial. Label the tube or vial with the CCoamppalnedtehoAlMd DfTor-aMn-al4yswiosrbkysheeleetctarnosdpartaiyacmhastos psapgeectorfossctoupdyy.notebook. VALIDATION 10.1 10.2 PQrueacliistiyonCoanntdrolAc-cunrotacayp-plnioctabalpeplicable 10.3 Other Validation Parameters- not applicable 11~ .0DATAANALYSIS ~ 11.1 None 2.0 ATTACHMENTS 12.1 Worksheet AMDT-M-4 130REFERENCES 13.1 AMDT-EP-22 Routine Maintenance of Ultra-Turrax T-25 4.0 REVISIONS Revision Number Reafosr Choannge Revision Dae 000146 00006 s Sa Simpie TL EF Worksheet AMDT-M-4 FCI C55 FCs Tare nd Ew EeEE] er P r r r r r a y r Tr rr ny rt r t r -- rrr ttt tr rr tt rt rr beebo -- 1 ---- ------ e rrr y PoeIolo iverSalugge rrr ee] _-- Plogex eee 7 e e e -- EE r r r e -- s r ] =--- | Fake pp------------------ mm ---- sm | Feat | e er r r r r r ] [Blowdown10seardrvnessESml wp, errs -------------------------- PAMTmofTTScnonprieig pg A ---- F Por gaite ee ee 77m mmmer ee ee] 000147 0008? 6 3M EnvironmentalLaboratory Method Analysis of Fluoride UsiTnogn tSheeleSctkiavlearElSeecgtmreondeted Flow Analyzer With Method Identification Number: AMDT-M- Adoption Date: /o-5.q Revision Number: 0 -_-- Revision Date: None Author: Deb Wri/gCyhnthtia Weber Approved By: up Leader 4 (2 ` a on Quality Assurance 1/95 Date 7-27-95" Date ASfoffetcwtaerde:DoIcBuMmeMntSs:WorAdM,D6T.-0EP-26, Operation and Maintenance of the Skalar Segmented Flow Analyzer 000148 000088 1 the Bormann 1.1 This methodis the Dohmann DfXor2t0h0e0an(aAlyMsiDsT-foMr-fl1u)o,raidned, ctohlelremcatleldyinexTtIraScAteBd fforroamnaslaymsptslewsituhsianng SloengmSeelnetcetdivFelEolwecAtnraoldeyz(eIrSEw)i.tThhIeSEa.nalysis is performed using the Skalar 1.2 Samples can be tissues, serum, biological material, or other materials extracted on 20KEYWORDS________ 2.1 Skalar, segmented flow, fluoride. 3.1 Follow standard laboratory safety practices. 4.0 SUPPLIES AND MA ERIALS 4.1 Supplies 44..11..21 4.1.3 ASuatmopplipeectusp,so,x4fomrLd oprlaesqtuiicvcaluepnstwwiitthhcpalpasstic Polypropylene volumetric flasks, 100 mL. tips 41.4 Cnaurmubreirdsg,e components, refer to the Skalar Methods for components and part 4.1.5 Sample prefilters, Evergreen 4.2 4R.e2a.g1enBtrsij 35.30% S.F.AS. Detergent 4.2.2 TISAB TISAB II Il buffer solution: Purchase TISAB II from add 2.5 mL or 100 ppm fluoride solution Orion. and 1 To 1 liter mL.Brij. of 4.2.3 4.2.4 Sampler rinsing solution: Dilute TISAB IT Nitric acid solution for decontamination, 1 1:1 with Milli-Q water. N (lab grade): Slowly add 64 tmhLesvoclonucmeenturpatteod1 nLitrwiictahciMdil(lHiN-QO,w)atte0r.250 mLs of Milli-Q water.Bring 4.3 4S.t3a.n1daSrtdosck solution, 100 ppm F: purchased from Orion. 4.3.2 Intermediate with Milli-Q swtaatnedra.rdU,se10poplpymp:roDpiylluetnee1v0omluLmsetorficstfolcaksksso,lution to 100 mLs 4.3.3 Working standard: Make up the following working standards by adding the "vooxlfourmdesoropfuimntpemramtedeiapitpeeorsr,sttooc5k0smtaLnsdaorfdTiIndSiAcaBteadndonditlhuetitnabglet,6 u1s0i0ngsaLs with Milli-Q water. [00p 00m pm7 p 5 eee nted -- -- SBeeemm 08pm p py p 000149 002 006.9 $1 Skalar Segmented Flow Auto Analyzer SansTM Sysiem equipped with ISE 6.1 Hinitgehrfceorenncceenst.rations of alkalinity, chloride, phosphate, sulfate or iron can caves 7-1 Swiatmhpilne3s0sdhaoyusl.d be sored in polyethylene botles. Samples should be analyzed 8.1 8P.r1e.p1araPtriepoanreocfalCiablraitbiroantsitoanndSartdasndasarindssection 4.3, 8.2 C8.a2l.i1braTthieonstandards are analyzed at the beginning of the run. 8.3 8S.t3o.r1a"gesSttaaCnnoddanardrdidsstiaarorenespsrteofproaerrdeSidntacatanapdpmaeriddnspiomluymproofpyelveenrey vsoixlummoenttrhisc,floaskass. nNeceewssary. 9.1 9S:ta1r.t1 9.1.2 UCplaPmrpodcoewdnurtehe pumpdecks, air bars and Put the fluoride electrodes in the electrode schaammpbleerr.-pump tubing. 9-1.3 Tbauthm. on the power of the sampler, pumps, offset potentiometer and heating 99.-11..45 TtPuuutrmnthoeonnretahtgeheenitn-itlenritfneaercsfe,ianccetohmebpeaufptoperrroe,prdtiihasetpelcaboyotmtaplneudst.eprri.nter. Make sure you 9.1.6 Let the system stabilize for approximately 30 minutes. 9.2 S9t2a.rt1inCgreaatRe uansample the name of the ftialbe,leanbdy sperleescstEinNgTFEIRL.ES, TABLE, and CREATE, type in 9.2.2 9.2.3 PDPirRaiInlNttTth,heeGssaRammpOplUleerPtsae1.btltTei,hnigissnstweoirlttlheedpraiipnnpttrhotephresiyeasnttteiernme trauubnl.emobyfbspaumsephliernsg,EnSuCm,ber of 9.2.4 FsFieWlcl/otnRhdeUssNfaoOmrpUslTaemcwpualpyeswwoiantshht,htehaesnadsmtnpanludemarbrdedsro,onsofatmspneleceeosdn,dtswoafboserheftishlelaeSdn.admpdrlief.is. IW and 9.2.5 Set the baseline. 000150 3 0007 . 9:2.5. smIeaSleyelcetbcietngiGnCRtOAhePNHDTaIRtCaOSHL,anPRdAEliNAnELgLTPaIanMneldE..puSIswfhiyitoncugh cFt7ao.ntnhoetAgneatlyrseials-Ptiamnee,lypoyu 9.2.5.b2aUsseelitnhe.e sAmdajlulstsctrheewbdarsievleirnefournttihle the bottom of the screen. iotfifssaetppprootxeinmtaitoemleyte3r/t4o isnecth tfherom 92.5. t3hCehienctekrftahceehsicgrheeswt#s3t.andard and adjust the gain, if necessary, with 9-2.6 GwtoiolrlteontouCtmObteNo TrtuhRne.OA(LnSaelPlyeAscNtisEoLrPa,dneeaslne.ldectot aannaallyyssiisspbaynelp.reDsessinegleEcNttThEeR.an)alByrseissstThaarb 99..22..78 SPerleescsttSheTsApRacTebtaorsttaortbrthienganuaplytshies.local menu. 9.2.9 paTrnyedpsesseEyleoNcutTrSEITRDA(fRionrTitcihAaolNisAc),eLstY)h,SeIcshSao.mopsleertuanbnlienwghwiicthhyooruwictrheoauttedthuendseyrst9e.2m.t1ab(loe.r 9.2. 10s0A!oftt1e0rtshaerrtiignhgttnhuemsboefrtwoafres,amstpalretsthaensdamtphlaert.heMsaakmeplseu/rweatshha/tatihetismaemspalreer is 9.2. 11Select GRAPHICS, REAL TIME (0 view the progress of the analysis. 9.3 9L.o3a.d1inaGgnodtasonedlCecOPtNrLiTOnARtiDOn.Lg SPtehAleeNcEtDLaA,tUaTp-rFOeisClseAtLheCsUpLacAeTbaIrOtNo barnidngenutperthtehelofciallenmaemneu(or 9.3.2 Thioghvliiegwhttthheecialliebrtoatbieonprciunrtveed, CURVE. agnodtporGeRssAEPNHTIECRS),. CALIBRATION 99..33..34 TToo prpriinntt tthhee lhoigwhlelveevellsccurreveen,, SEPTINGS. Chang the max ppuusshh EPSRCINtoTgSeCtRoEutEoNf.graphics. Select 5 valu 10 approximately 900. Go to CAL 9.3.5 pCRreUetsuRsrVnEENtoTaSnEEdRTpTarneIdsNsGPESRSICaN,nTdacSnhdCaREnngEteeErNt.hePtrmoeapsrxsinPvtaRslIuaNempTblaSecCkpRetaEokEs4.0N9,5, go to EDIT, 9.3.6 TOUoTpPriUnTt.theSerleesuclttsPRgoINtoTCERONfoTrRtOheLEpPsAoNnEoLr,PSRPNAfCoErBtAheRL,asOerU.TPUT, 9.4 9S.h4u.t1doPuwtnall the reagent-lines in Milli-Q water. 99..44..23 AoLffefttseertthetphosetyessnyttseitmoemrmeitnhesares.frToirunsamepdporfcofoxmtiphmleaethteeealltyyi,n3tg0ubmmaitnohuftofentsh.weeseakmepnldesr., Lpeuamvpe alniqduid in 99..44..54 Tthaekelintehse.electrode out and soak in 100 ppm F overnight, Release the pump-decks, air bars and samplerpump-tubing. 99..44..76 SOenleFcrtidFaIyL,ESw,mproefsfstAheLcTomFpuatnedr,sedleicstplQaUyIaTntdoienxtietrfatchee pforrogtrhaemw.eekend. VALIDATION. 10.1 Q10u.a1l.itRyuCnonatsrionldard (mid to high concentration) every 10 samples,If a asicgcneipfuicbalnet csthaanndgaerdinwiplelabkehuesiegdh.t Tochceurrse,maonilnyintghesasmapmlpelseswiblelfboerertehaenallayszted. 000151 4 Ov 000071 10.2 P1r0e.c2i.sSioeneaMnedthoAdccVualriadcatyion Report number AMDT-M-8.0.V1 1100..43 ORetfheerrto VMaeltihdaotdiVoanliPdaatriaomneRteeprosrt Number AMDT-M-8.0.v1 1L0D1A1.T1 ACAalNcuAlaLtiYoSnsIS ~~~ 000000 1111..11.. 21ATlhlecsatlacnudlaatridocnusravree better, idsopnleotbtyedthbey StkhaelSakrasloafrtswoafrtew.are.should be 0.995 or 1111..32 `PWwrrieitphtaearltelhpesapScrtkeuaaddgysehnedueomtcbsuemtroensotunsmtmahaenrdpirpzilneatcodeuattisan,. tIihnneictilsautldu,eddyastafeomlptdhleere.pvrMoianltkuomeueta,, cawonepdiygbhoitnfsdtuhtes.oegdetehtec,r osnutmomsatruydyshdeiestkaannddwbpaeckiunptodtihsekss.tudy notebook. Back up all data and spreadsheets 11.4 E1l1e.c4t.ro1niGcLDPatsatudies: Electronic data is copied onto the Study floppy disk for each 11.4.20sttuhdeyr, astnuddiaelss:oAdlaltdaaitsaciospcioepdieodntoontaoflaofplpoypdpiysdkisthkatthiastsitsosrteodreind tinhethleabl.ab, 2.0 ATTA N' None 13.0 REFERENCES 13.1 DAXM2D0T0-0M-O1rg,anTihcerHmaallidEextArnaacltyizoenro-LfiFvleuroride by Means ofa Modified Dohrrmann 1133..32 ASkMaDlTa-rEMPet-h2o6d,s,Op#e3r3a5t,ioSnkaalnadr MMaeitnhtoednsanMcaenoufalthe Skalar Segmented Flow Analyzer 14.0 REVISIONS Revision Number Reafosrchoannge Revision Die 000152 3 000072 3M Environmental Laboratory ~ Method Thermal ExtractionofFOrlguaonriicdeHablyidMeeAannaslyozfear M-oSdeirfuimed Dohrmann DX2000 Method Identification Number: AMDT-M-14 Revision Number: 0 `Adoption Date: /o-3-7 Revision Date: None Author: Rich Youngblom Approved by: rouop rLe)ader lDa0t/e5/ 2S El GIZA Quality Assurance J Date Software: MS Word 5.1a Affected Documents: AMDTE-xMp-an2daFbllueoroindeAMneaalsyuzrerement by Meansofan Orion EA940 AMDT-OErgPa-n3ic RHoaultiidneeAnMaaliynzteernance of2 Modified Dohrmann DX2000 000153 1 0000" 0 SCOPE. APPLICABLE COMPOUNDS, AND MATRICES f1l.u1oSricdoepef:roTmhivsarmieotushomdatrisicfeosr.thTehoepefrlautoiroindeoisfatyDpoicharlmlyacnonllDecXt2e0d 0in0TwIhSeAnBitsiosluutsieodn tfoorexatnraalcytsis `1w.i2thApapnliiocnasbelleecCtiovmepeoleucntrdosde:. Fluorochemicals or other fluorinated compounds. 1.3 Matrices: Biological fluids, particularly serum. 2.0 KEYWORDS 2.1 Fluoride, fluorine, extraction, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, DX2000, fluorochemicals. 3.0 PRECAUTIONS 3.1 3.2 Glassware Glassware aisndfraegxihlae,usbtrogkaseensgclaasnsbmeaeyxtcraeumseeliynjhuorti.es. 3.3 Pressurized gases, proper compressed gas handling practices required. 33..54 PSootlevnetnitalbbaisoehdaszaarmdpsledsuematoythfelasbhi,olmoagiycanlemeadtrtioceasl.loUwsteheapmptroopdrriyatdeopwenrsboenfaolrperostteacrttiinvge run. equipment. 4.0 SUPPLIES AND MATERIALS TMI water 4.1 4.2 Compressed Compressed Oxygen, Helium, HHiygdhroPcuarirtbyonGrfardeee,, regulated regulated to to 30 45 PSL PSI. 4.3 Quartz glass sample boat with TeflonTM tubing, Dohrmann 890-097 or equivalent. 4.4 Quartz glass combustion tube, Reliance Glass G-9405-012 or. equivalent. 4.5 4.6 OSraimopnle94c0ol9l9e9ctTiootnavliaolns,icHDSPtrEe.ngth Adjustment Buffer (TISAB 11 ) or`equivalent. 4.8 Polystyrene pipettes. 4.9 Activated Charcoal, E. Merck 2005 or equivalent. 4.10 4.11 MHiasmcielltloanneSoyursinlgaeboorrateoqruyivgalleansts.ware 50 EQUIPMENT 0000000 5.1 5.2 RIoBsMemcoomupnattiDbolher3m8a6nonrD4X8260c0o0mpOurtgera.nic Halide Analyzer, modified for fluoride extraction. 55..43 EDxXc2e0l 0S0presoafdtswhaereet,, vveerrssiioonn 51..000,ormogdreiaftieerd for fluoride extraction. 6.0 INTERFERENCES 6.1 Sample size is limited to approximately 100 ul, This may vary from matrix to matrix. 000154 00007 2 7 LE HAND 7.1 Samples are to be handled with plastic pipettes. A new pipette i to be used for each sample. 8.0 CALIBRATION AND STANDARDIZATION 8.1 Preparation of Calibration Standards 8R.e1f.e1rTthoeprsottaoncdoalrdfsorrtehqautirpreodjefcotr.each project will need to be appropriate for that individual project. 88..11.32 TFyoprircaablbliyt5s0e-r5u0m0sptupdmiesF,C-us9e5beienfmsetehraunmolassttahnedmaartdrsixa.re used. 82 Calibration - Overview `cTuhrevenoirsmraelquciarleidbrtahteiponroicsetdhuereflluiosrtieddebceulrovwe i(sAuMsDedT.-M-2). However, ifan optional spiked serum 8an2d.1coAmbcuasltiibrnagtitohnecmurusvienfgorthtehesaDmXe2m0e0t0hoisdsgeannerdamtaetdrbixy tsyppiekiasngthseamspalmepslewsittho kbenotewsnteds.tandards s8.p2i.k2edT.ypically, tree replicates of cach standard and five concentrationsofstandards will be R8e2c3oveSrteadnFdar(dugc)uorvnethwielyl-abxeisp.lotGteenderaastMeaassreSgrpeiskseidonFc(uurgv)eoanndthcealxc-ualxaitseatnhde eSqtuaantdiaorndfoMratshse line a8.n2d.4thMears2svaSlpuiek.ed F (ug) = (Amount spiked in mL) x ( Cone. of standard in ppm) x (0.6004) 8.2.5 Standard*MFaCs-s95Reisco6v0e.r0e4d%FF(tuhge)re=fo(rTeI0S.A6B00v4oilsutmhee finacmtoLr)usxe(dOtroiocnonrveeardtinFgC-in95pptom)F 8.3 Calibration - Procedure 8833..11.1StRaurtn U2por more Clean Cycles when starting instrument cach day. More clean cycles may be used if the previous samples contained high concentrations offluoride. 8833.22.1BlParnekpasre sample using the same methods and typeofmatrix as the test sample. 8833..22..23 PFuotr sraebrbuimt sbtluadnikesi,nuDsoehbremefansnerbuomata.s tChoemmbautsrtixs.ample as described in section 9.0 and 8an3a.l2y.z4e Fsoarmprlabebaitccsotruddiiensg,ttohememtehtoedr rAeMaDdiTn-gMf.o-r2a fbolratnhkesiaomnpsleeleschtoiuvledelbeect0r.o0d3epapnamlyosrisl.ower jbuedfogreemeprnotcoefedtihnegopweirtahttohretchaelirberaadtiionng. isBstuamblesawmiptlhersesupneclt tthoishilsitmoirticiaslrreeaacdhiendg,so(rpurnetvilioiunst4h8e h8o3u.r2s.)5. For non-rabbit studies, the blank readings should reach a predetermined ion concentration 8be3f.o2r.e6prItocmeaeydibnegnweictehstshaerycatloibmraitxioan.pproximately S0 mg ofcharcoal with the sample to aid combustion. 000155 00007 3 8.33 Standard Curve d8.a3y.3(.=13I0fbmeLe)f.serum is frozen, thaw at least enough to complete the standard curve analysis for the 88.33..33..32 SPtiaprtetwtieth10th0euLloowfebsetefstsaenrdaurmd cionntcoeDntorhatrimoan.nnUssaimnpglae boat. Hamilton syringe, eject a fixed quantity of the standard or in thebeef serum. on or in the matrix. For rabbit studies, use 4 uLof standard and eject it on 833.4 may be uAsteldeaasttt3herdeipslcicraetteisonsohfotuhlde baenaluyssetd. for the lowest standard concentration; more replicates 8.33.5 8.3.3.6 Combust the sample Runall 15 standards. asIfdoensecrriebpeldicianteseisctsiiognni9f.i3caanntldyadniaflfyerzeentacfcroormditnhge to AMDT-M-2, other two orlepdlirceaptleisc,atreunanadnotthhaterthseamnpelwerfeoprlitchaatteswtainldabred.useIndditcoactaelciunldaatteaththeartetghreesnseiwonrceuprlvieca.te replaces the 8le.a3s3t.07.95W,hceonnsaullltswtiatnhdasrudpserhvaivseorb.een run, calculate the 12. r2 must be at least 0.95. Ifitis not at c8h.a3.n3g.e8d.ANneeww ssttaannddaarrdd ccuurrvvee mshaoyuladlsboebreunruwnhaetnthtehedicsocrmebtuisotnioofnthteubaenaolryssta.mple matrix is 8.4 Storage Conditions for Standards T8y.p4.i1caSltloyr,asgtearnedaqrudisreamreensttsorfeord sattarnodoarmdsteamrpeedraetpuernedeinntploansttihcesicnrdeiwvitdoupalbostttlaensd.ards used. 8.4.2 New FC-95 standards should be prepared at least once a month. 9.0 PROCEDURES 99..11.1TyCpoimcbaulsOtpieornattuibnegtCeomnpderiattiuornes:= 950C. 99..11..23 OVaxpyogreiznatainodnH/Derlyiiunmg flow time = = 50 cc/minute. 240 seconds. 9.1.4 Bake time = 300 seconds. 9.2 Start Up Procedure: 99..22..12IOftpheenptrhoegSrYamSiTsEnoMt sStEarTtUedP, swtiarntdtohwe.EOX program on the PC. 99..22..34 PCultostehetfheumSaYceSTmoEdMulSeEaTnUdPthweicneldloiwn.the READY mode. 9.2.5 When found in the tCheEoLvLenChHaEs CreKacmheendu.the READY temperature, run the CLEAN BOAT program 9.2.6 See AMDT-EP-3 for details of the Dohrmann software. 99..33.1SaOmppelnetEhxetSraAcMtiPoLnEPrHocAeTduCrHe:and pipetie 100uL of sample into the BOAT. It may be dnoencee,sscahrayrctooamlisxhoauplpdroaxlismoabteelmyix5e0dmign wohficlhearecstoaablliwsihtihngthtehesabmaspelleinteo aanidd cwohmebnusgteinoenr.aItifngthtihseis standard curve 9.3.2 Close SAMPLE HATCH 000156 4 00007 9.3.3 Add appropriate volumeof TISAB solution or 1:1 TISAB:Milli-QTM water mixture to a labeled sample collection vial. Typically 0.6 mL to 15 mL are used. For rabbit studies, use 1.0 or 2.0 mL of 1:1 TISAB:Milli-QTM water mixture. 9.3.4 Place the vial so inches. Gases released that the during tip of the pyrolysis COMBUSTION TUBE must bubble through the iTsIiSnAtBh.e TISAB at least 0.25 9.3.5 Run the EOX-WATER program found in the RUN menu. 9.3.6 When the EOX program is finished, remove the collection vial from the combustion tube. 9.3.7 Ifundiluted TISAB was used to collect the sample, add an equal volume of Milli-QTM water to the TISAB to make 1:1 TISAB:Milli-QTM. 9.3.8 Rinse the endof the combustion tube with Milli-QTM water and wipe with a KIMWIPE to remove any TISAB remaining on the tube. 9.3.9 Open the sample hatch and remove any remaining ash from the boat. Ash can be removed * `witha cotton tipped applicator and/or vacuumed out. It may be necessary to scrap particles off the bottom with a spatula or other similar device. A drop of Milli-QTM water may be added to the boat to aid in the Clean Cycle. 59.331.110 CRluonsethteheChLatEchA.NBOAT program. 93.12 Sample is ready for analysis by ion selective electrode (AMDT-M-2). 9.4 Sample Calculations 9.4.1 Use the standard curve to calculate the sample value. 9.4.2 Sample Mass Recovered F (ug) = (TISAB vol in mL) x(Orionreadinginppm-intercept) (Slope) 10.0 VALIDATION 10.1 Quality Control 10.1.1 Daily Start Up Check Samples: Once the standard curve is established, each day of analysis is started by analyzing QC samples. The QC samples are to be the same as the lowest concentration spiked samples usedto generate the standard curve. Each concentration must be done in triplicate unless the first two replicates are within 20%ofthe standard curve, then a third replicate is not necessary. 102 Precision and Accuracy: See method development analysis and sample analysis in Fluoride Notebooks 2,3, and 5. Precision and accuracy varies when analyzing samplesofdifferent `matrices and different reference compounds. 10.3 Other Validation Parameters: NA 11.0 DATA ANALYSIS 11.1 Calculations 11.1.1 For the standard curve, use regression analysis in Excel, version 5.0 or greater. 11.1.2 To calculate the fluoride contraction in the sample, see method AMDT-M-2. 000157 5 0000 11.2 Analyzing the Data a1r1e.2w.i1th2in m2u0s%tobfeeaatclheaostth0e.r95anodr tghreeaotuetrl.ie"rOuits lgireerast"ermtahyanb2e0e0x%colfudtehdeifavtewroagoef otfhetthhorseeetrweoploircates nleostsedthiannt5he0%FinoafltRheepaovretraagleonogfwtihtohsethtewor.easAonnyitswuachs ocuotnlsiiedrsersehdoualndoubtelipero.inted out in the data and 12.0 ATTACHMENTS None 13.0 REFERENCES ~~~ 13.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer Operator's Manual (Manual 915349, revision B, December 1993) 13.2 AMDT-M-2 Fluoride Measurement by Means of an Orion EA940 Expandable lon Analyzer 1A3n.a3lyAzMerDT-EP-3 Routine Maintenance ofa Modified Dohrmann DX2000 Organic Halide 140REVISIONS 00 Revision Number ReaforsChoannge Revision Dae 000158 5 ronnOo 9.3 Quality Assurance Unit Statement 000159 000079 Attachment D GLP Study Quality Assurance Statement Study Title: Single-dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits Study Number: AMDT-042095.1 Name of Auditor: Kari Rambo This study has been inspected by the Quality Assurance Unit as indicated in the following table. The findings were reported to the study director and management. EInospmection DaTtees Phase 11-13-95 11-13-95 Final Report MDautnegIonsmpeecntionSRteupdoyrDtierdectioor 1-13.95 11-1395 J A : J L=1355 QAU Auditor Date 000160 000050 2 9.4 Key Personnel Involved in the Study 000161 000081 3M Environmental Laboratory Key Personnel . Thermal extraction followed by analysis using Orion ion analyzer: Jim Johnson . Deb Wright ! Rich Youngblom Deann Plummer `Thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Documentation and Reporting: Jim Johnson Rich Youngblom Quality Assurance Unit: Gale Van Buskirk Cynthia Weber Kari Rambo 000162 000082 9.11 Data 000163 000083 9.11.1 Summary and raw data; ug F" in whole liver as determined by thermal extraction followed by analysis using Orion ion analyzer. 000164 000054 Summary of Combustion Data - Liver AMDT-042095.1, HWIT 6329-159 As Referenced in Final Report section 6.0 DATA ANALYSIS Total pug Fluoride in Whole Liver Mean per Dose Group per Day Sacrifised* Control Group, Day 15** pg Std. Dev. 27.1 + 3.1 Control Group, Day 28 38.7 + 16.3 100 mg/kg dose (T6246), Day 15 (0.12 mg/kg)*** 37.4 + 7.9 100 mg/kg dose (T6246), Day 28 (0.12 mg/kg)*** 29.6 + 1.3 500 mg/kg dose (T6246), Day 15 123.1 + 14.0 (0.60 mg/kg)*** 500 mg/kg dose (T6246), Day 28 100.5 + 5.9 (0.60 mg/kg)*** *Calculated as a mean triplicate samples from each of two male and two female rabbits. **Only two livers analyzed from this group. ***Test material is a 0.12% solution of FC-95, actual dose in parenthesis. 000265 000085 rovry (WW) (WM) (grams) (grams) (ug) (mglkg) a bor 0 as Torta oo on wa TToorroeer:s oSioon m Sie aaa ReTisaLLS pont spike-1 91% 1.26 oh0.110 spike-4 97% 1.04 0.142 Danza in Sie m0 Pager 000166 00086 Fess pAcotmuFal- ApveermaFg-e liver Wlhiovelre TotwahlolF-ein % infiver inliver bumed weight ory (WM) (WW) (grams) (grams) liver (ug) Dosage (mak) FFssii007799-21 112107 118 0011527 88311 wes EF5s4t07a9t3 113368 00114514 7e3210 Fsns1ie63 11336 135 0011507 772200 e450 FeFst41i122:12 115454 146 00115384 997788 1429 500 Fs4501716-213 116490 0o1t3s9 798718 FF5s4s0o7766..32 118718 176 00111244 77811 134 500 FFesaatt1iaa1z 113234 13 0011435 8911s5 217 50 FF5s4a0t8i4-a1d 11.4528 0o13r4 6o6s4 FF5s4i008844.32 11.7638 185 00112400 6666.44 1104 500 FeFsdd0000ee.21 11.3495 149 O011375 770900 173 500 FFes4i008078.13 01.64010 00113205 7e9a07 FFssii008877.32 0033862 036 00113315 88377 315 100 FsFsii008e6-t2 014.833s 0765 O01s34 88433 ses 100 FsLIiVoEsReBsLK 1 00.245892 00.113644 843 LVVEERRSBPLKK12 89% 0110352 00.115288 LLIVVEERRSBPLAKN2K 111% 012.4303 00..114209 lLiVveEr RblSanPkIKES 111% 013.5275 00111313 FFs5i4008800.-21 00330048 034 00115573 771188 283 100 FF5544008090.-31 00359012 0o1u3r9 872118 F5F450409990.:32 00.540444 0483 00115480 a8221 7 100 FsFasattitaazt 00337327 03% 001135%7 881100 200 100 FFSs4a11010.413 00338637 00113535 784100 FsFaS140100.023 00.348217 0400 00113s4 774400 206 100 FF5S44008800-12 004.7850 03% 0011322 775544 288 100 54080 0132 0103 75.4 Page 2 000167 RPTISILXLS 000087 Foss Actual Average Whole Totailn % pnplmvFe-r pipnmfeFr- bulimveerd weliivgehrt wlhiovelre Dosage F54052-1 ory 0W.W53)7 (WW) (0gr1a1m7s) Gr7a8m4s) (ug) (maka) FF5544008022:.32 00.446333 0478 00114358 7788.44 374 100 FF5544110066--12 0o8s3s9 063 0134 0125 823 823 522 100 BFl5an4k1063 0.5% 0.181 0121 823 0.133 LUvveerrssppiikkee21 8821%% 017.5095 00.111538 UUvveerrssppiikee3d 8756%% 01..84977 00.116548 LLivveerrssppiikee 65 6760%% 11.3460 00114572 fliivveerr bbllaannkk--12 a05s49 00..114373 fliivveerr sbpliaknek633-1 103% 012.7088 00..111403 lliivveerrssppiikkee 6633.-23 19569%% 11..1619 00.113423 lFiv5e4r08s8pi-k1e 63-4 101% 01..8415 00.114058 843 FF5s4d00888e--23 00.645915 054 00112332 884433 458 100 Ifviveerrsbpliaknek-643-1 117% 012.9302 00.112336 ivveerrssppiikkee 6833.32 8888%% 00..992722 00.114379 verspike 634 90% 0.910 0.149 RPTISSLXLS - 000168 000088 9.11.2 Summary and raw data; ug F" in whole liver as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. 000169 000089 RE: 6329-159 LIVER SAMPLES AMDT 42095.1 Date of Analysis: 5-23 and 5-24-95 Analyst: DOW Co "alas Stata Quta. The samples are burned in the Dohrman at 950 C using between 8.1 and 0.2 grams of the liver. The gas is collected in 2.8 mL of 1:1 TISAB/Milli-Q water. The samples are then analyzed on a Skalar Segmented Flow Analyzer using the lon Specific Electrode (SE) Method. TISAB buffer is added to each sample as it proceeds through the system. The sample then goes through a heated mixing coil before the potential between the ion selective electrode and the reference electrode is measured. The signal is amplified and related to the fluoride concentration. The instrument was calibrated in the ranges of 0.815 - 8.15 ppm and 8.15 - 1.50 ppm fluoride. The standard curve for the high range was plotted using the inverse logarithm option. The standard curve for the low range is linear. All standards and stahemspelecsurwveesr.e Atllhernescualltcsublaetleodw b0y.0t6h0e1Spkaplmaraspopfetawraorne using the raw data as #.###%, A quality control standard was analyzed every 10 samples to check for accuracy and drift. Raw data is taken from the appropriate calibrated range of the Skalar printout and summarized on an Excel spreadsheet. The final results are adjusted for the collection volume and any subsequent dilutions. 000170 000090 SUMMARY of 6329-159 LIVER SAMPLES AMDT 42095.1 000 "hi\as Skaler HEa E072 000035e 2200 oODur I o0s3s0 05a 2 002230000 m30306 274 FBwBrsIs 0o0m2 2200 oomrnias 00332 020507 21588%7 FE00BBL32 000022 2200 0o1u0s7l5 00331% 033 swss 22593% 268 EFlSzI 000;2 2200 00105057 003ds 03% 8663355 0au%st 312 DGoRsOeLUePvl1:0 EFlSSsOTaA 004;5 2200 0o1u06n9 08338 06634s9 6n56d7 EE00732 o00m5 2200 oownssL 00745 319 swoese 6403% 2 E Bo2l 00on 2200 oowzs 0033s 03 m2m55s 23100s8 23 FBo0s3s 0002 2200 oouos 002%3 s0e95s 2138967 BEo08s2s 000026 2200 o0u0s 0L0y isl 0S09355 o23d9o0 458 DusGcRLOUe0Pv04emglh:s EBOOME2 000085 EBoowss o00u5 FEsi0ss23 0000s5 EE0092212 000056 BFiunla 000086 EE006632 000046 EBo0s2s 00005s BF0o8a5 000m4 FEB200003 FE099912 000084 FF0l93 0000s BFsllGsz 00003 FEla9s2 000033 F093 003 000171 2200 ooonll 00&519 2200 oousnt oLsn 2200 oOHL 00668 2200 oolles 00% 2200 00118% 0036s 2200 oonmie o0n%l 2200 ooluess 0oms 2200 o0iBs o06s3 2200 oouuse o0s% 2200 0o1n0ss3 0033s 2200 0011hs 003599 2200 0o1l2s o03k% 2200 Oo1n9s 0033% 20 ous 04s HWI1SSLSUM 0s msan 2 oa 47 8B2a 1a6s47 Lol 8s2i%a 5s3m36 853 075 7H3e6 Ssells ss w6il 740637s5 om Bs0a1o 5M8B76 es 075 883266 705607 61 n88m2e6 2593m9 od nMmmee m 281si 0% m82a1n6 046235 4595 We1ss 831219 03 w80s%6 23104 OLS 4030 TTeessss 1247s 299 Bes nn page 1 000091 SUMMARY of 6329-159 LIVER SAMPLES AMDT 42095.1 F SR LSAT Sap AG Arig To Gar ergs F081 003 FFs8i008032 0N0D3 20 20 01322 OLB 046 030 03 7542 saz 3491 3 2- 20 01028 ND 52 ND FFs00R8-2l 001122 FFU0L8L3 0010l9 FFSISlSI2 001008 FF007912 OOIlL FFsSs0l7l9e3l 001130 Fle 012 DoseLGeRvOeUl:PSS0mgkg FFS4L1623 001123 FF182232 00113S FFS0077612 0011S5 FF0U7L6 00123 FFS1I3B23 0011 F861 Ola Fsi0842 016 FFs8i049831 001133 F082 018 F098 013 20 20 01595 0166 Lad 148 88965 1840 146 8895 11s 130 2200 001120s7 1L4a7 88889635 118042s 2200 O01M36T0 11530 Le B88233 110256306 19 20 OISTL 146 wu Dus 20 20 0OM1006 1L4R6 148 BB1177 10160671 13 20 ous 17 ne 128 20 20 00M1003 LLees 16 7791692 L1l2l0s7 12 20 0130 182 1824 177.98 2200 OOIsS8s3 1L7s 1&2 897482 117850245s 178 20 ons 222 78058 17359 20 20 00113463 223453 233 7788005588 118839.6650 182 20 ol L6s oLs2 15108 2200 0o1n2m7 LLSo 181 9911522 117.30276 166 20 0IN8 200 6642 138% 20 0142 220 218 6642 1876 las 2200 oOMw6 21923 67684925 11122033 20 01469 1S8 1s8 7895 1833 136 20 ops 2m 895 16723 000172 HWI-158L.SUM Pag2e Ceeey2 tssns DaiOpatesrssotfoanrhtesAtnesay0s0199o5u.0r5.:23 1s0:4a0 szsar lysFiliNasme. CISKALARDATAVWIOATASLIVSEZ3RAS1 2F sorT ome udom owtire woww Ss 5SSmat mwsz ooonmws 0O00o2 uuwoow 37SSSiaaawwdases oooimns 0o 010s mowmw BSo aSSiwaaese? 0o1%o0 0o1bma lwooowwm Sg2 HW6ammmTaamn lioa alitemo womm 85 6 aoeakka: aaaooos B12o0 ooowwmnne ooowwm n2io eFraesraosrs:s aao6ee ll1oo0 ooowhmnme ooonnl aaammmeee nwnw 2a rrFoeostss 0oo0ee2 Ll1oo ooooieme ooonnx mwmiawrr mmxuwu 2EE2 Fsoawost ae ai1m2 ww Fr 2ZBa reioss & Bra 0ao0s22 L0o oooiwwmss oawnne ssseewse auwsnoe Paget s25u%rip S3 pei 14 sx s25t rFrsoeorrsz aiions 23Bo oochummm ooonnn mmmeeew wuaisnm 55 FFFreeios hirae0 3B30 SocNuuns odonne mmmmeesm nnieun 3F SrSosA s Ppris noe km ne 4pBao easis iioin 23B0 oaoemmms oowmnn mes we 8b"ooosama ooaiinee BBER oouwme- sien oGGmooii aaGmwm aaonss ooomwm wbieemr P ghLa ame ioonnn BBBooooweme iiinnn GGGloeii odGmem aoannn ooonmn iuuemm gBS8au0 oaWna ow oa1onm wm note a 2FP LE oE moeek2 oio0i0io2 2FBB0R oou0-1w6m00 02) iinn oGGlooii aGawmm ooosn omoomn ennwee i3 eFSosaoamss oaorne BB30 oou0wmmm tilann mmmess omea et Gm ans om me 5G8o rrFraaimmmaas iaoon 3BBoodkuhm aiinn mBmames nmmeaen 228Ga eoSnmne ootenn ww 20 ome in men mx 22 8a eranm: aann n3 otowen dakn mRmw oEmaa 3 oo ww isnxs o%8r eFFoosneaess: ooonnn 0TBo oooihums 6ila mRneewms mmmnmn %3 fDpoeeel xooiu I BI oSOEm 4iinn rwWeemahnaw B[[I CrNe ou = ooum om onm ome j 7jita]o ooounu 0IIomonus wiimn mmneoenwm miiawe BB 8 rFDns ooounn BfBoooomom iloanm bNmeEomeibnen o 588 fTFaoamsmesa oonn nBLoooutommm lmnnmoummsaE bweiane gB&EroEm . otne om Boom momo 3Boha85 epToweemns o0oonu0n0 1LBooooowme oauhnm mhmnew awbena 5% frrooeonnms ooonnn BBBooSdtEa ootnnh Waunmnowonme %%EoFTwoss we ioomun om 3noGem o|w wNRm wowu FN - 2 3 g pana mcr a L,g rineAN J He trN womsA iwem n IH I 5SE 0Er H H E Bin f Q pli girgmenmaa aotibe opW 0oe 3i 835nujeaeomml poio5xem 3FiBl omeees 0o[0x 3 re oo SB2zi 3 oEsenK1 LE05o0x8 3 2i:27 o8wau%sh 0o>0x0 g 5% Swe i wBN Boougsmmw wooxcwE oe p5apoe2ony0e 2 BBoossem2 anoo2 m 2B20 os0I11eZ33 31M w ywe w Io owe ranatw ommomoogm6m00 wtyn wmee Go se woatsmooonies w2eoawn ow om tweoemddy: oam o28 0om58 saz a35 nIeennmzaaraciss ooosmms 2000 ooomumera ooownm 553 VSSaeeneesss oooornns BBWoooouomumem iiimmn oGGweo aabmmm 7F 5 SSehsaP om on oem in Go Gm f0Doo rFFreisim:s ooouuu 2BBoouhowmne ooemmn wwwnuk wmheee lFa rsrheera oo0um BBWo ooomoomanm oiinnn oGwhs bomw S fo SSueewss ooommm BBWoo ouodmem in on oGwet obmm Sghh8 oaSomae ae oodonw om oom om 388aas=aaa ooonon B2Booooommme o11nmb oGme obwm FF5ai esas oooion BB3ooouommms ooanm am wm oim ooww 5&5 rFfrrriimom:ms oooooo 3BBooommm ooonnx nammmme bmns 28&8mFoams ae ootont om Boum 0% Bean 2E2E55R rom Hana ooonnnt 2BBooommws ooonnm mwmmwe ennnne ~ Page2 aoarwms ooommm wubwmme ow on mw oaasnk oooomnn uimmem an on um oanns oown mumm oonw oomn ummm [rr & rFFeoonoenr ooomnn 2330 0oommmmee ooowmn mnmaoamw mmmusn T77o Frroeosrrs oooann 3BBooonommnee oaomwn nomaaemo noaunnn B 7HfESous wm o=fm e Bom im an ow 8cg 35 3 33 & rons 995-05-23 15:26 Output of : 950523A1 oftware : version 6.1 c1990,93 perator vow ate of the Analysis : 1995-05-23 10:18 nalysis File Name + C:\SKALAR\DATA\HWIDATA\LIVERS\950523A1 BS etwe luoride 1.5 = teres sogsion 1ope run parern a + = corrected value of the sample esult = 10 s s = Slope of the electrode 2P5=an-20.e0000s%00 lHuhoaritdietLn order = 2 `orrelation : r = 0.99847 esult = a2 * x? + al * x + a0 FSS 2 = ~0.00000 i= 0.00024 ampler TVTSayeaspmhm'pepelre TTiimmee :i|| fS5102A0100ss0eecc. E aSpkeeciawE lp Soinnegle meals Height | Joan Suter TAneaetHdtleoSnKeiFTgaEhcttRoEr 1:| 881000s mm ml RDeosatnapins runs i0i1 uFosperrocies"e | mo me Q Abrk 420\9as51 329-159 Zi 000179 000099 995-05-23 15:26 OutPut of : luoride 1.5 Path number : 3 Signal type : Debubbled Decolor i Yes system Number : 0 diLute i No Resample : No ddiilG Touhtrpeusthold :: 04095 Window event : Off sl sTandard : Ignore s2 sTandard : Ignore ss34 ssTTaannddaarrdd :: IIggnnoorree s5 sTandard : Ignore $6 sTandard : 0.150 s7 sTandard : 0.300 $8 sTandard : 0.600 $9 sTandard : 1.200 O$r1d0er sT: aInndvaerrs:de Loga1r.5i0t0hm Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed StArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 Filter i No Rfoergmeunlearatio: n : No output BRT luoride L Path number : 0 Signal type : Debubbled Dseycsotleomr Number : 0No diLute : No Resample : No dil Threshold : 4095 diG output : 0 Window event : Off 950523A1 000180 000100 995-05-23 15:26 Output of : s2i sirtaannddaarrdd:i 0o.o01m5 3S3i STraennddarad 1| 000.800% 3S3e SSTtaannadaarrad: 0o.n1s2o0 s7 sTandard : Ignore 3l85o srSTtaaannnaddaaarrradd || ITIogonnneeorrrsee ODSttriradmiregeerenesriv:oanlL2uiem:itPPM ||: 1S08000 SDuec PNSteaaarktshiiafgpnheoorree : P102.o0inteSsdeecc Hoasura window | 75. FRfieolgrtenenirearationes. l: eyNRoo cutput. | aren 950523A1 000151 000101 995-05-23 15:26 os Typ tiw wa ss12 ss3 ss56 0 . ss87 23 Ss310 sioaw 5Tuou 33 oouu >Loouu 2 3 ouou i5 uou 57 aow 3Jouou J[a u 2 3 oouu iPouu [sa ou 3Voaw >Cuou D3u [Ja ou Ptouu viuu vaw Duu Ident InitialTrWaacserh DWriafsch Ssttaannddaarrdd 12 sSttaannddaarrdd 43 SSttaannddaarrdd 65 sSttaannddaarrdd 87 stSatnadnadradrd 109 DWraisfht BBLLKK 12 F540B7L5K-13 FF5544007755--32 FF5544008811--21 FF5544108015--31 DWraisfht FF5544110055--32 FFSS44110077--12 FF5544017047--13 FF5544007744--32 FF5544008855--12 DWraisfht F540BL8K5-31 BSLPKK 21 SSPPKK 23 SSPPKKS& SBPLKK 16 DWriafsth SBLPKX 21 output of Fluoride 1.5 PEM Ch Result F Time 33 01..046702 21635 33 01..046705 368257 33 00..006731 971460 33 00..018088 11026940 33 00..112574 11464126 33 00..621815 11976952 33 11..426318 22311440 33 10..503620 22742930 33 00..008746 23081345 33 001.007747 33139616 33 00..006795 33574117 33 00..006653 34809615 33 00..006687 44243185 33 10..056206 44585312 33 00..007700 45914115 3 0.866 5291 33 00..007823 55466450 33 00..007608 55892302 33 00..007608 66314625 33 01..057105 66581912 33 00..008607 750942i5 33 00..007721 77323136 33 00..115112 77578434 33 00..110145 87093158 33 00..113006 88217415 33 01..058303 88769139 33 00..006890 99104310 3 00115 93s : 950523A1 Fluoride L pon Ch Result F Time 44 0s.u5s5a5n9 00 44 0#.w5a5n7e6 00 44 00l.0012s5s5 00 44 00..00851379 o0 44 00..11510819 0 44 00..23581356 o 44 00..55515828 00 44 0#..5a6%6#4s o0 44 00..00532574 00 44 00..00339237 00 44 00..00323017 00 44 00..00220101 00 44 00..00115898 o0 44 0#..5#6#5a4% 00 44 00..00224210 00 4 0.4378 UskWigh see 44 00..00520774 0 44 00..00128330 00 44 00..00212704 00 44 00..05282308 00 44 0#..045#8%6 00 44 00..00224845 0o 44 00..10496844 o0 44 00..10088600 o0 44 00..01920218 00 44 00..05464655 00 44 0#..0#2#1a88 o0 40.1076 GOOANZ Page 1 of 2 tsits cashshs poowz2 995-05-23 15:26 OutPut of": 950523A1 Fluoride 1.5 py Fluoride L Poy os Typ Ident Ch Result F Time 45 oouu SSPPKK 23 33 00..112140 99647905 iu SPK 3 3 0.118 9845 7u F54082-1 3 0.124 10020 33 ouou FF$544008822--32 33 00..112284 1100139760 LJ ouou 2a FF5544111111--12 33 00..100938 Drift 3 1.512 1100574260 10834 FSa ow FS41w11a-s3h 33 00..01640 1111123335 sou F54079-1 3 0.124 11420 5Tuou Pou FF5544007799--32 FS4116-1 333 00.1111195 0.138 1111579750 11944 35 oua FF5544111166--23 33 00..112376 1122121985 IFuY FFS3e4111122--12 33 00.113500 1122644750 Sioau F541D1r2i-f3t 33 01..143373 1122892915 5 5 oowu F340W7a6s-h1 33 00..105804 1133233437 T3 uou FF5S44007766--32 33 00..115319 1133659270 >5 oouu FFS544111133--12 33 00..112283 1143034773 L 2 ouu 3 ou FF5543101834--31 F54084-2 33 3 00..112835 0.157 114323271 14571 5Pouou FF5544008948--31 33 00.113363 1144794271 T5wd DWraisfht 33 01..046500 1155302975 3 oouu F5544009988--23 33 00..113493 1155464271 3 ou FS4087-1 3 0.077 15796 2Louou 3 ou FF5544008877--23 33 00..007753 F54088-1 3 0.123 1156915492 16322 F5a ou FF5544008388--23 33 00.1008812 1166363762 5Toaw Dwraisfht 33 01..0468)5 117304759 ry Runout Wash 3 0.060 17321 Ch Result F Time 44 00..01903153 o0 4 0.1085 0 4 0.1151 0 44 00..11210442 o0 44 00..00875731 00 440.f.5#6#3#3d 0o 4 0.1017 0 4 0.1149 0 44 00..11002842 00 44 00..11311351 00 44 00..11321179 00 44 00..11436060 00 440.h.5#5R3e7t 00 44 00..11456021 0 44 00..11313366 o0 4 0.1134 0 44 00..11313394 4 0.1530 00 0 44 00..112333%0 G0 44 0#..5#3#9#3% 00 44 00..11332870 20 4 0.0384 0 44 00..00334038 00 44 00..10413440 0 44 00..05455276 0a 44 44..248434% Page 2 of 2 000183 0101 BESTCOPY AVAILABLE 000184 000101 BEST COPY AVAILABLE 000185 000107 ------e ee A342 oF GOEIIN PE EE a Fiugride 4.5 ----m-- Ee | 1 pd i i] 0 Iii os vo i i i IH w hl 26 i i od ! ol | i 1eq PP oe i he R<A < Y 1" lg i i Y1 i] Lowha RW aan i |B aT TTT TTT TTT a SEEAL Fisbaln | CreltoRAitpa1s - 000186 000105 REE ee TT gerry Tr ye a -------------- Rau d3t2 oF 3TSIiey "3s Fluoride 1.5 --me------------------ 3! % | i I 1 i | 4| 0 il iT i | Za 1 | It 1 I | :} i | Hi i i 1 "3 w "No i i i ww ne civics EVIE RAY 24 ve aan L} e PVIR R fed fhiii five seen TEC |Freein Caan en TTT ~TCOPY AVAILABLE 000187 000107 a wi 1 Ray data of 95032364 Fluoride 43 ------ w; # ; i 1 li | I!| |Pl| ; i } 2 ele Pa ETATaeall CaGw,atC eattT? T 0 0e T L4La FE ghVaVnUaRIAG GR i UR E TRg L TL EdLh ya cm me TTT TEs SEAIt Fase G3 riieEedmist BESTCOPY AVAILABLE 000188 000105 ee Bau dts oF 80SZE Thride 45 Pi - ! & aw ! i 41 | P=Rs) AL ba Ple waTgE ! &Nw 3 JU el CL i | fijeSea e1h g VT ` FV V ML BT TE A BESTCOPY AVAILABLE 000189 000109 355999555---0050-552-4224 12:48 o utPut of + 9 950s 24A1 00 "fs, |las Amp 4204571 oftwars : version 6.1 c1990,93 329-159 )iven porator cow BESTCOPY AVAILABLE ite of the Analysis : 1995-03-24 07:50 nalysis File Name C:\SKALAR\DATA\HWIDATA\LIVERS\930524AL alluiobrirdaetio1n.5order = Inverse Logarithm i10pe cs = paren f[r=e=i] xG1:= ccoorrrreecctteedd vvaalluues ooff tthhee ssaemmscleentration 1 ssult = 10 s 4 s = Slops of the electrode 2 = 0.00000 1= 0.00065 0 = -1.22861 Luoride alibration order = 2 `orrelation ; r = 0.99392 ssult = az xx ral rx sao 5212=1 0001.00001000320410 angler TNSyiapnmenos Tine |iis50a100s0ec. GRarsh TTiimmee 1| 120 sseec Tsapkeeciaulp :| SRoimnsgle needle Height : 70 mm. Luter AaniisTleaidttliieoonnHeiFvgaohclttuomre:|| 8100 2.5 mm mL ReDislauntoiioen runs ii 1 uRseesrrocfsisls:i No RE 0001390 000110 = 5395-05-24 12:48 Output of : ple ae luoride 1.5 SPiagtnhalnumtbyepre : De3 bubbled disylsutteem Humber :: 0to Resanols : Ho W4di1il6ndooTwhuretesvpheonulttd :i: O04f0f95 s5i2 ssTTaannddaarrdd :: IIggnnoorree 5S3i ssTTaannddaarrdd:: IIggnnoorrse s5 sTandard : Ignore sSS76e sssTTTaaannndddaaarrrddd ::: 000...316050000 55510 csrtaannddaarrdd : 11..250000 ODridmeerns:ionI:nverPsPeM Logarithm sttrairgtgerValLuiemit :: 1505000 SDUec PSteaarktsihgnaorpee:: P60ointaSdec oMNedasureigwnoirnedow:: 17520 S%ec FRielgteenreration :: dNoo ofourmtuplaut: #4.### `luoride L SPaitchnalnumtbyepre :: D0ebubbled Desycsotleomr Number :: f0io iRetsuatmeple : tlioo d4iilG Tohuretsphoultd:: 40095 Window event : Off 950524A1 BEST COPY AVALLABLE 000191 000111 9935-05-24 12:48 Output of : sl sTandard : 9.015 52 sTandard : 0.030 5s34 ssTTaannddaarrdd :: 00..005900 s5 sTandard : 0.120 56 sfandard : 0.150 s578 ssTTaannddaarrdd :: IIggnnoorree $9 sTandard : Ignore s10 sTandard : Ignore order : 2 Dimension : PEM tstrairgtgervalLuiemit :: 5108000 DSUec Peak shape : Pointed start ignore : 60 Sec eNd ignore 120 Sec Measure Filter window :: N75o % Regeneration i: No ofuotrpmuutla :: cF4.:#=#c#3F 950524A1 000192 000112 P 295-05-24 12:48 os Trp Ident i3iw InitialTrDWaracisefhrt "sSzi ssttaanncdaarrWadash2l S3i SSttaannddaarrdd 43 235 sSttaannddaarrdd 55 i0L 33S79e SSStttaaannndddaaarrrdaa 97 5iSoaw$10 standarDdarisf1ht0 THI u 554400902321 osIuu 35F544410009662213 Fta u FS406B-L3K i sFI ou SSSPeKK123 HTEw DWriafsch oosuuu SSSPeeK d65 I3u0 LLIIVVEERR BBLLKK--1Z iSuu LIVESRPCB6L3K-31 isSouuu SSSPPPKKK 666333--432 S5 w4 Darisfht Douu FP5S4i00083s--17 5 Fi 340B8LK334 siuu SsPeKK6633--12 TiSuuu SSPKKL66i33.-l43 osdu bBrizf: Lu Vash E2R ou SSPeK-z1 output of: 9505241 Fluoride 1.5 Fluoride pe een Ch Result F Time Ch Result F 333 10r..i0456s35 333 0001.1000878739 33 0001101900 33 00.1115257 333 100.l.2s23l374 333 011...044589925 33 00.0088s4 333 000..000887s85 33 00..008505 333 000..00190155 33 10.1405345 333 000...111534254 33 00..008756 33 00..016078 333 000...111150008 33 01..045334 33 00..008875 33 00..008890 33 00..112075 333 000...101080555 :3 01..048872 3 0.059 33 00/.111025 3236h15 444 0os.laasdsees: S75i32z56 44i 00#...00421e86s20s 11206825 44 001.00963013 1164132 44 00.l11511604 211197368848 444 000...742577211223 222373128258 444 #00L..e88k37a51R00 320813i8 44 00..00556042 333331465170 444 000...000384671417 33781833 44 00..00413138 4i4204461135 444 000...000778685044 44388300 44 0#.l8e5e1s1s 455912313931 444 000...111423768390 53466333 44 00..0031456 33899123 44 00..008186%2 666531136s23 444 000...100948359108 6562332 44 0#.l8n7a0e7 77201180 44 00..00354271 77536350 44 00..0021187 779714s2 44 00.l018i4672 S6e20a65d822 444 000...00088144579 S6671541 4i 00.102064790 S023 4 seen 99311473 14 00..0089407 Time 0o0 000 00 00 000 000 000 0000 000 00 000 00 00 000 00 00 00 00 000 00 0 000193 page 1 of 2 222 " 000113 " 995-05-24 12:48 OutPut of ': 950524A1 Fluoride 1.5 Fluoride L pon pom os Typ Ident Ch Result F Time Siouu SBLPKK-33 33 00..016174 99646991 T5 ouou F540S8e9K--13 33 00..007847 109803195 35 ouu FF5544008859--23 33 00..007743 1100139638 opuu 3F344009995--12 33 00..007778 1100574148 52aow DWraisfht 33 01..048579 1110180573 4HSE uu FFFSS5i44ll0ll9as9---lz3 333 000...000787224 11111l52s94l44s sTuu F5S4i1ll0s5-13 33 00..007741 1111796480 >5 oouu FS544110099--23 33 00..007761 1122219155 ELR u FS544008800--12 33 00..007745 1122644655 3iaou F340D8r0i-f3t 33 01.4.88A0151228991983 3 oTww RunOut WWaasshh 33 00..005599 1133148658 Ch Result F Time 44 00..00317696 00 44 00..00311688 0o 44 00..00321889 00 44 00..00339819 00 44 04..8454740% 00 444000...000324066313 000 4400..00320585 00 4400..00233454 00 4400..00330361 00 44 #0..#8#5#7#8 00 44 8#. .4p6a8nFe 0 Page 2 of 2 2 2, #Z , 000194 000111 2 Calibration Swrvs of SESE Fluoride L mmm i 7 |i 5.001340 Ea ee neoprse oapan mee n) & [oR ' Cwm BESTCOPY AVAILABLE 000195 000115 ee Salibration curva of S508248: Fluide 15 -- -- -- EERTee ETSI eT gl ii : / | + 0E30528 oe a eee eee} to = dar iwverse Logarithn mr BESTCOPY AVAILABLE 000196 000116 5 ee dg mats of HOE2001 Fluorine 1.5 eee i i i : Poa 31 itl cu i wo ! tii i 2 i 1 |1 HdHE | H 1 :i <n oH Pal oOa ial Pa ; eS Fh \L a 3, i i ;2 ; 3s | | CIE yon 3, Py ' A I A 4 fil `7! ` 0 Tire canny samEl FremainEerisaems BESTCOPY AVAILABLE 000197 000117 5 1. | : ] ! i dae H a o C { m! %% 1 ee [soe 27| of VER UV of WU i 2 oe ee ee = E 582 TeR n E eTTime --_--_--t Sm BESTCOPY AVAILABLE 000: 198 000115 . ---------------------- Ra dziz of 305243 Fluoride 4% -------- wr " 1 | { 1 \ | } Ls 5% SS ! | pases ee Ye agar g BIVOSZAAANT CARAT pl ag CE | alo|CeLl-eRmaeiaila 1 TTT BESTCOPY AVAILABLE 000199 0e119 20 9.11.3 Summary and raw data; ppm F in serum as determined by thermal extraction followed by analysis using Orion ion analyzer. 000200 000120 HAWMIDT6320492-019559.1 Dohrmann Serum Analysis Analysis Dates: 04/28/95 - 05/10/95 All serum samples were thermally extracted by a modified Dohrmann DTIXS2A0B00soOlrugtiaonni.c HTahleidseamApnlaelsyzweerreanmdeacsolulreecdteodninana O1r:1iomnilEliA9Q4w0ater and expandable ion analyzer. The Dohrmann was calibrated using 32ppm, 63ppm, 126ppm, and 253ppm FC-95 standards. The Orion was calibrated by direct measurement with no blank correction using 0.05pprm, 0.1ppm, 0.5ppm, 1.0ppm and 1.5ppm F" standards. The slope, intercept, and correlation were recorded in the appropriate logbook. A summary table is included, showing the ppm F- in each sample (see page 2r)e.adiTnhgeovfableuleoowf 0".N05D." has been entered for any sample with an Orion Pages 3 - 10 show the excel spreadsheet that was generated when the samples were analyzed. Pages 11 - 18 show the same spreadsheet with "ND" inserted where the Orion reading is below 0.05. 000201 Page 1 of 18 000121 reason OoumseOmgty Fosczoena.anttssriGo5n Sample ne Shou t2hes oevew dates Bfaaaess w N No N wNo N m mw p wMw fPfaaaems NN Now NNNoo mmmwow no Bfaizs wNoo MMow mNw M ow OoguSmokg Supe ano ow tates ener fmBeoomrm Nw woo wM oN NMww wNp Bffoeewro N N No N NMwN Mn NMN ffaem NNw NNmMN OowgeifOmgka Sangh dhow her tones Zena fffaoarom owN NO ww NM wM NN owo ffFaeammee N N Nw M Mmw M M Nw o Nho BFaasw wNoo Naos M wo N Oowge 00makG _Sumpe dhew Show rahe iShew ines hows Ons Omz osm fBfeooamor NNNoO owwNo MN wN wNM N Mm N mMw NNM Nw Bffooaawmm N NNoo MNNiNsMNM N1 NMw N mNo w NMw Nw wo FfFaaiwee NNNOoo M M WM w M N mo MN 0 wNmMNNonwo Ouse S0Omso Samp nos how 2b 2ebow dabews Days Seber Ow owas fFmoaomss 13N13o M w is M wm112 1132 iosw w1e3 M mN w FTFiaocnsa 1N1%M3 Mw two wNNw 13312 11r33a 311s 3113 ww BBFaaaem Zdowoee sw ae2 wmaes 31dso 171143 8 riez Haow ww 000202 000122 2 60201595.XLS STUD#Y 6328-150 384 mLGf Cone. Actual * Quantity |DI: 0am 5 z SoFlCutIi5o solFuCtIiSon rMeeadtienrg s(ammpolre TfIiSnaAlB tc%ury = pAoctmuFa-l spMiaksesd Mroavstsd STaerruom Sblaa1nm0ki.2 spiked (ppm) (00pp00m230%F (g00r.1a1m00s) vol22(00mL) luglug)ins50a.7m420ple (ug F) C(0u.0g074F20) sseuummssppikkee 6633-21 00.000048 6633 00007700 001100 2200 9932%% 1144 0O1155 001144 sseurumm ssppiikkee 994455--12 00..000084 9944.55 00004683 001100 2200 4S1e%% 01832 002233 0001932 sseerruumm ssppiikkee 112266.21 00000044 112266 00.10959 001100 2200 16051%% 2340 003300 003210 sseeumm ssppiikkee 112266.34 00..000044 112266 00.1135 001100 pi 0040 010 222000 8958%% 2360 0.80 003300 002360 0.080 FFrrieairosras Fratose 0000322 001100 2200 0025 010 20 00..4685 00..008658 051 0081 rSratones 000713 001100 2200 0.1647 0.e0s87 aao2s Sunes 0033 001100 2200 0084 010 20 7629 00792 17 017 sauoncss 0000338 001100 2200 007759 00..007795 sSaurmsmpotn Sari spk 00000044 663 0000688 001100 2200 7903%% 1114 001155 O01114 0004 6 0073 010 20 9% 15 015 ois SSaramm sset 00000044 112266 001110 001100 2200 774%% 2220 003300 002220 SSemnssoi?s 00000044 112266 001111 001100 2200 7723%% 2222 003300 002222 pSrainon 0000004: 112266 000129 001100 2200 787%% 2230 003300 002230 FSroniiamaSsps 10 Fsione.s 0004 126 0001405 001100 2200 68% 02510 030 00.20180 0027 010 20 054 0.054 FFesairoornas Faaoass 00002215 001100 2200 0020 010 20 00..4429 00..008892 0.40 0.040 FFsreiorsaeoss Frionna 00002210 001100 2200 0018 010 20 00..4412 00..008812 0.38 0.038 om tn 21 oma 1 0000228 001100 2200 00.5488 00..008589 0004 63 0071 010 20 94% 14 015 014 Foramc a 2 Frerons 000s 63 00002722 00100. 2200 95% 01.445 015 00.10445 0020 010 20 0.61 0.081 FFroerron.as 00001176 01i002200 00.3342 00..003324 000203 001001122:3 3 sszsressis STUoY8 sans 2384 | `FmiLvoef Tov `Conc. io `Actual a "Quantity ToDInE"e I abi ed I ions B~n solutio soTlutTion Croeooaadtitensg t(0me1Lo0nr wTf3rin0aEly orcevry p(pr0im5mo2F-y sSpiekesd GrcovrHadd JpTeoannnss o0Goo0nr0 oo0t1lo0 33be0e o0oi%s 0eo0or%m a Tgroes GGcooonrm 00o11l00 3330e0 o0o3sne 0o0.0o08m% pprors" cGGooosml oootllo 333e9e ooosnss 0oo0oo8rm8 pppan-- oocotomsn o0o1ll0a 3336ee o0o3sz8 r0O0oe5i ppTroroens o000o07 000111000 23300 ooin)n o0i.o0x8r1 JoTSitoisnet oGSsoas 00o11v00e 3b10e8 2TI5oH ooolex rTvaioennsst GcWoooww oo0d1lo0 333e90 o5is0s 00F0Y3)8 ppPnind oooooom 0oo1t0to3700 odoioas GGGooosis poonarss o0ooo0tnss o0o1ti0 02b0e a0o33n0 00o.003o300 joan-- ers oGGooonwr ooolil b33ee0 ooBezs 0a000o6%e -- Tpoer 000000si 6&3 oooooomm 00o1d10o0 Z32o00 tSomes aGvneh 0e11p5 0eo0nre6 pJpooo5i40i77-8 0ooG.ooo0t2:rs3 000o.11l1000 2303.000 0o0o.s34o6 o000..o00043m368 oreoernss oGGoootttss 0oo1tl0 d23e0 00=32 0Go0oo2sm Joheosnd occoosne 0oo1ll0 d37ee6 0005%%0 o000o03m30 posoesnss ooooomtms 00o11i00 3Z36o8 oo0s3st2 Go0o0osan 000204 00000123 4 cazmnsssas STUDY # 328.150 3847 Cn 5 rFeeronaness8 rraaoiryns8 Taaaisroeonrnsss FFFeaaarrresss8 JroTeorairsaa arooenres0s -- --1 jmFearsnsesy FaHotoior1ra2ss rliotreensa FeFrlaaoirorns12 Fraaeroneornssz reiirvooernnss FrFeaaorrrrsii1zss rarararoiro:a FFaasnoes: #5407812 sUFoGCoSH. Skea sCoFolCnuSei::on foo rAMecatoduiwanlg (ppm 1) Qsi(aamnmtoirety (rams) TfSDinAaIlRE| Tro%ny vl (n) gla) inoAsouumis:s a(MnaokseFsd (MreagvsEss B00o00a1sr7 0oatl0oo -- 222000 05V3s4 o0o0o3sn4 00000032312 aaalllooo 223000 o008i82s 00000088s83 0a00o02a7 a0ol1lo0 232000 a0037s64e 0a00.880s7 0aa0o20s4 o0ol1i0oo 252000 aoo'ssssa 000000816ss8 oo0os0ds6 00o11o00 222000 aoissaes 0a50808 00o00s2ss 0a01l10o0 2300 20 0asss i 00a00880ss oo01zs7 00a11l00o 322000 ao3sz4ss o000z334s8 00000ss 66 00o00o78m0 0a011l00o 222000 ssxx a11s43e o0i1s5 oo0s1n9s 000000m9 0oo1it0oo 2200 20 000543329 00000053s39 0oo02oo0 l0al1oo0 2200 20 0004331s7 0000008331s7 0a0o00:ss 0a011t00 2200 30 00o57s08s 0000005570s 0000003:ms2 aaollooo 2200 30 0ao3dn8d oo00oo5nt8 oo00ioms oo0l1lo0 2200 20 000442ss9 00000042s89 00o00o1s oa0ll1oo0 2200 20 oo0ssx 000.000838132 o0000e2s0 aa0ll1oo0 222000 0003301 000..000334210 o0o0i72 aalloo 2200 0033s4 00033s 0.014 0.10 2.0 0.28 0.028 000205 000125 = 63201595 XLS STUD#Y 6320-159 38411 [i FTaaoiore1tz FFaadoonnraiizz FFraaaoontz0i1zz FFraaanrnoennzz Frsaanoirae:z FFarnanoinnzz FFsessooss8e1r2z FFeassot70e.n12 Ssoumma2t SSoomm Ssppict2 SSaarruumm SSokkta SSearruumm sSp5kc8s SSaannaamm sspkks? FFarsaorross2z4e FFssastoon.2244 FFaaaaotsoeaz2ss F[ri)oraze sBeac 12 wwhhae 2 ispiekes4 Bkaankss Faekdoss 26 FFsaasro0a02264 FFraasoos7e.22e0 Fssoo.24 | FmCeS5r CFoCnOe5. AMestaelr Qsanmptley TTDIISABE%p| Acwal Mass Mass ssolpuitkieod so(lpuotmi)on (rpepadminF2g) ((gmrLamosr) voflin(amll) (urgcv/rgy) inpspammpFl-e s(puigkFe2d) (ruogvrd4 00.002275 00.1100 22.00 005514 00..005514 00001199 001100 22..00 003378 00..003378 00001178 001100 22.00 00.3336 00..003336 00002216 001100 22.00 00.5421 00..004512 00002272 001100 2200 00..5454 00..004585 00003486 001100 2200 00.9619 00..009619 0000442s 001100 2200 00..8847 00..008874 00004473 001100 2200 009845 00..009845 00003303 001100 2200 00..6608 00..006608 00000044 6633 00004304 001100 2200 4524%% 007687 00.1155 00.006779 00000044 6633 000055 001100 2200 7626%% 01918 001155 00.1110 00000044 6633 00008533 001100 2200 17110%% 1117 001155 O01117 00000044 6633 00005453 001100 2200 7635%% 01913 001155 O01110 0000421 001100 22.00 00.8414 00..008414 00001157 001100 22.00 003305 00..003350 00001168 001100 22.00 003337 00..003337 00003113 001100 2200 006217 00..006217 0004 63 00002323 001100 2200 43% 00.6453 0.15 00..006453 00000044 6633 00004474 001100 2200 6538%% 008385 001155 00.00888 00.000044 6633 0004 63 00.005427 00.1100 0051 010 2200 6682%% 20 67% 01.094 10 00.1155 00..01904 015 010 0000230 001100 22.00 00..3782 00..003732 00002119 001100 2200 0oa3t8 00..008318 00001176 001100 22.00 003332 00..003332 0000185 001100 2200 003311 00.00311 (001226 000206 L 63291595XLS STUDY # 6320-159 3 384 n Feate02n24 FFassio0n7n1..2244 FFsaas0o9952..2246 ssFasraorni.ozse FFssioosn0e-z2e4 FFeidoor2e6 FFsadoe0i90.22e4 Fraasdoomn02zss FFaseo1n092.2244 Fseoamtibaiake1 ssoomm baikk 21 sEariomespnc 2 FFaai0r2i.a2s6 Frsesdoosnaez2e6 FFsaaot79e.2s6 reFssoaniaasne FFeasroiseasss FFasisoornas.asna Fssaarrinaansea 5F4s0a8r2t1n9i2e FFssiaoors9a.i1s92z FFssiacnaiseitonan FFsssaotet2a3a6c0 FFssia0t9r.a30600 Fsso7a.260 mLof Conc. Actual Quantity = DI: "i ig | soFlCut3i5o spiked soFlCu9ti5on (pom) rMeaedtienrg (pom F) s(ammLpolre (grams) TfIiSnaAlB vol (mb) rov%ry (ug/g) pAcptmuaFlin sample sMpaiskesd (ug F) Mfoavsisd (ug F-) 00001134 00.1100 2200 00.2258 00..002285 00001250 001100 2200 00..3400 00..003300 00001143 001100 2200 002276 00.002267 00001112 001100 2200 002234 00..002234 00001235 001100 2200 00.2560 00..002560 00001134 001100 2200 002277 00..002277 0000113s 001100 2200 00..2268 00..002268 00001146 001100 2200 00.2372 00.003227 00001165 001100 2200 003312 00.003312 00001383 001100 2200 003727 00..003727 0004 63 00007218 001100 2200 4% 01545 015 00..01545 0004 63 00006842 001100 2200 108% 1163 015 001135 00006535 001100 2200 1113 o01n3 00006614 001100 2200 1123 001123 00005684 001100 2200 1132 001123 00006868 001100 22.00 1134 0011a3 00007637 001100 2200 5 13 00..1153 00003518 001100 2200 1182 001182 00008850 001100 2200 176 00.1178 00.006733 00.1100 22..00 0067 010 20 11.35 00.1135 13 013 00008787 001100 2200 115a 00115a 00006655 00.1100 2200 133 00.1133 00004633 001100 2200 01.837 00.10837 00005843 001100 2200 013 o0r1t3 000207 Ceo1e7 63291595. XLS STUD#Y 6326-159 384 | io} FFesiaotseeas6t0o Sonam 1 Sonam 2 Sonam pk 1 Serum Sok 2 FFssue01s12.002222 FFsis0i719e.002222 Fse082.028 FFss4a017n9.002288 Fse116.028 F54099.4 F5F6s0409999-.182 Foi000.24 Fests Fsien102 Fsi0s8.360 Faiz Fsaize Fsaiziz Fsa1220 Fsaizas Fsaiz152 Fsati2360 SERUMSPIKE 83.1 SSEERRUUMM SSPPIIKKEES63323 SSEeRRUUMMSSPPIIKEEZMs212 sSerEuRmUbMaSnPkIKEM63 serum bark spsiakrea6b3a1nka spes32 spa633 spa634 spiasss "pin36 spike 637 spagia SeSeurmumbiinke2t Semapie 63-1 Soumspike 63.2 mFlC9o5f CFoCnIeS. AMcettuearl Qsuaanmtpiltey! TIDSIABT % Acwal Mass Mass Ssolpuitkioe.d so(lpuotmi)on (rpepadminFg-) ((grmaomsr) vfoilna(mlL) (rucguiruyg) inpspammpFl-e s(upgikFe)d (rucgurFd-) 0.063 010 2.0 0069 010 20 Ta 0.14 1.4 0.14 0025 0032 010 010 20 20 0.50 0.65 0.050 0.065 0004 63 0004 63 0.069 010 0076 010 20 91% 20 101% 14 15 015 0.14 015 015 0046 0041 010 010 20 20 081 0.82 0.091 0.082 0040 010 2.0 0045 010 2.0 081 0.081 0.89 0.089 0048 010 20 003 010 20 0.96 0.10 071 0.071 0033 003 010 010 20 2.0 0.66 073 0.066 0.073 0028 010 20 0026 010 20 058 051 0.055 0.081 0074 0087 010 010 20 20 15 17 0.15 0.17 0047 0033 010 010 20 20 095 067 0.095 0.067 0029 0038 010 010 20 20 0.57 0.76 0.057 0.076 0059 010 20 0066 010 20 12 0.12 13 0.13 0051 010 20 0086 010 2.0 1.0 0.10 13 0.13 0058 010 20 0041 010 20 1.2 0.82 0.12 0.082 0004 0004 63 63 005 010 0079 010 20 78% 20 105% 12 16 015 012 015 0.16 0004 63 0.004 126 0074 010 0083 010 20 20 97% 55% 15 17 015 015 030 017 0.004 126 0.004 126 042 010 042 010 20 77% 20 79% 23 24 030 023 030 0.24 010 010 20 0040 010 20 2.0 079 0.20 0.079 0004 63 0019 0058 010 010 20 20 77% 033 12 0.15 0.033 0.12 0004 63 0004 63 0058 010 0082 010 20 77% 20 121% 12 1.8 015 012 015 0.18 0004 0004 63 63 0053 005 010 010 20 20 70% 73% 1.1 1.1 015 015 0.11 0.11 0004 0004 63 63 0088 0072 010 010 20 20 77% 95% 1.2 1.4 015 015 0.12 0.14 0004 63 0060 010 0045 010 20 20 79% 1.2 0.0 015 0.12 0.090 0004 6 0017 012 010 010 20 20 153% 033 23 015 0.033 023 0004 6 021 010 20 273% 41 015 0.41 . 000125 000208 3 ssaotsss xs STUD#Y ss29.150 asa | "FmClSo5f CFoCnIc5. MAecttuearl' Qsuaamnptlitey TIDISFAB [11] % Aci wal Mass Mass T Ee sSolputiioessoluo tpioomn (pomroeaatdinhg a ((r0ma1Lr0oer) vfo2oinp0all )bS recv(r)yco&rpsn0epaa4mymg2eiF-e(esepiBgke)d M(0r00oav4Ere2dL FSoont Sa oG00oMoss&a aooommsea oo0l1fo0 2z2o00 sses9%ww 11G383 o0oi1isss oo6n1nss Ta ray ay 0o 000 s6ea GaOo Gos oooddooo 222000 so9om%w 111442 00o11l3s5 oooniness rb orrw n aos 6 cocaaomm oooidloo 2z70o0 sok o014i41s 015 00oo0n0se8s forined fora coowte oor 0o1t0o ono 222000 000435s7a 0o000o33t74 bToiaome o0o0r0 oo 0o1o0 oto 777600 00304 032 00..003460 0032 JToeonsnea rece ccaome oollo 2706 oats ol 20 ooess 000044sd 036 0036 r--t fron ocoo sa cooei sow 001100 oto 7200 20 7708%% 0G32z8 12 ois 060132 ois ons ja r ma: oo e30Go0s oo o0ot1lo0o 277006 ex 00183222 ots 0o0ns6 0032 SSboeinonnma;s aa0oo0ss &6 Gooomgles 00o11l00 22z00o s7ow%k vu1a4 0oo1ir5ss ooeunnree Troemriinoe Jos ocaom 001100 7700 ome odo 70 0o3s0 00005%2 03s 00% Fneardoonn caaos 0a1l0o 00s oto 7200 70 o0s7e 0000%% 08s 006 ToTrooooronrst ooozo 0% ooddoo odo z7o6 10 o0a5r2 0s 00002812 0043 Toroims Ton GGao Gam 0ot1o0 010 2300 70 0043s5 ot 0000004a33 TJoia ez coa ao oats o00d11o00 2260 26 003300 039 000.000348900 rroay od Gcootwe Gone o0t1o0 010 2200 70 00338 039 00.003387 0.039 ro Gos 010 20 038 0.038 F54087.28 000209 0018 0.10 20 0.35 *0.035 00012. 9 6329159S.XLS STUD#Y 6329-159 384 lot7 Cone: Atal QUAY 11 DETTE rr SoFlCutIi5o soFlCuIti5on rMeeatdienrg s(ammLpolee TfIiSnAalB 're%wy Apcowaml Msapiskesd Mreavsss TE Rua ne LD den ID Spiked (ppm) _ (pm F-_(grams) vol (mL) (ug/ug} in sample {ug F-) (ug F-) CCFG5nn4h1wo1m4iw4m2mm8ceeeessss 0oo0ddMdd 855 00o0.000o1%k7 0000.11110000 2333.0000 4ss1an% 0ooG.ee3Fss3 01e13i5 0Soo.o0ni3u3 wSroroeonssecotsns y 004 6 oGGaoomon aooltloo 23266s ohn 001523 oie coooonen wSSwneaanwsmmsss o00o00d0 s56s 00o080o1 a0ol1d0ao 33z6o hmmnww G1133a oooinnees aoenas Troeaa ccoazm ootlo 1z6e o08z 0G0o8i2 00H0210 00012" 0 7 63201555XLS STUD#Y 6329-159 SERUM 384 oFCOf5 CFoCnS5e AMcetutaelr" Qusaanmipltey DI TISAB % Actwal Mass Mass Serum Bl[ian1k ssolpuiteiod so(lpuotmi)on (rpepadminEg-) 0.036 ((gmrLamosr) 0.10 voflin(amli) 20 (rucgv/rgy) inpspammpFl-e Wo s(upgiked (rucgurd) sseerumm sbpaknek263-1 0.004 63 00007200 001100 2200 93% N1D4 015 014 `ssseeeruuumm ssspppikkkeee 699344-..25512 000...000000444 99644355 000.000467630 000.111000 222.000 945216%%% 011..8342 000.221335 0000.118342 sseeuummssppkikee 11226612 00.000044 112266 0G01959 001100 2200 16051%% 3210 003300 003210 sseeruumm ssppiikkee 11226643 00000044 112266 00.1135 001100 F54081-4 0040 010 2200 988%% 20 3206 No 003300 003206 ND F5454017057..44 0000322 001100 2200 NNoD NNDD 7F55441007645--44 00002354 001100 2200 NNDD NNDD BBllaannkk 2| 007314 001100 2200 61sa 0.648 BlBlaannkk 34 Blank 5 000838 001100 2200 0038 010 20 7127 007127 ND ND Blank 6 0035 010 20 ND NO SSSeeerrruuummm SSSpppkki-.c123 000.000000444 666333 000.000675835 000.111000 222000 79930%%% 111.541 000.111555 000.111451 SSeerruumm SSppii 00000084 112266 001110 001100 2200 6774%% 2220 003300 002202 SSeerruumm SSppii? 00000044 112266 OO1111 001100 2200 7723%% 2222 003300 00222 SSeerruumm SSopdk 0000004 112266 0001928 001100 2200 677%% 2203 003300 002230 F5S4e0u8m5S.p4k10 0004 126 000.4105 001100 2200 68% 2N1D 030 0N2D1 F5544100846--44 00002275 001100 2200 NNDD NNDD F554408037.44 00002210 001100 2200 NNDD nNoD F545049140-04 00002210 001100 2200 NNDD NNDD s5e4r0u9m1b-i4k 1 00002199 001100 2200 NNoO NNDD sseerruumm bsipkk21 0004 63 00007218 001100 2200 9% N1O4 015 0N.O14 se5r4u0m9s7p.4k 2 F54102:4 0004 63 00007222 001100 2200 95% 1N4D 015 0N1o4 0020 010 20 ND ND F5F451400781..44 00001167 001100 2200 NNDD NNDo 000211 000131 Ul cmrss ns STUD#Y 8329-159 SERUM A 3 FCS5 FCCoOeS AMatSer OsaRmpEle TEIDSRAB % Acawual Mass Mayss T Toros B SolutioE solution rGeaodilng l(ae mrLeeor fa i3npa6l a|Crocywy pDaeFp - spoeim Dkee)d ranehwS)rd TTFoooeriiioooanrosss GGcooonsn ooollle330 I>] -> TTToooernttiaeoyse oooooorm ooorlle b33eee Ivb3d] I"-]a TFFooorrroovnaoenness ro0or0m a0r1e0 3366 -Io]d --Jos FTTootvooyna0nse 0ooo0ln%s 0ao1rt0o 333666 oIo] I] TeFoorioeearsse GGoooon ooaeit 333600 -- o-~n Tounetieses Goooroo ooorlle 333 2TT3ae oootrnee TFFooorvnotntnesas co0ooemr 0oo1tle0 33388 22w 0o1n% TTTooroowrorrsaa cooooonms o0a1vre0 333666 fo S-Io] BFFooeiioorraeesss GGGoootnss oo0t1io0 373666 IJvoI]] - Tproieeonletn) ooGoonmw a0o1el 333600 II]] +3 TSiurmmoeok2 000000i 653 0000003s%1 oo0t1lo0 333096 1msa% 1oIan oSt1s2 oSiuas FfTooeiracalonieass coGoobn:s oo0v1te0 3136h6 I-~] I- TTTooeioveoenr s Tens ooGoootns ao0n1ie0 333686 o~d I--] oor 010 36 i] I] FTToeiiroonanns-8B 0oo0ol1rss 00o11l00 311888 ->o --I] FFoioooainnesa GGGooonss 0o01t10o0 333688 =- a3 000212 A 000132 12 s3201595.XLS STUD#Y 6329-159 SERUM 384 in F7554300936-058 FF5544018170--88 F5544008898.-88 5544100860.-88 5544100992.. 88 7F5544017164..88 F5F4sa018121--88 F5544101739..88 F5544009884.-88 sFe5r4u1m16bi-k81 sseerruumm bslpk 1k2 se5r4u0m75s1p22k FF55441008511122 F545047140.71122 FF554008865..1122 FF5544007867--1122 5F544008943.1122 F5544019001-.1122 F5544100927:.1122 5451400787-.1122 F545049037.11122 F5F45049150.11122 5F450419160..1122 F75544007980.1122 mFUC3G5r CFoCnIe5: AMcettuearl Qsuaamnpiltey TIDSIAB % HEActual Mass Mass ssopliutkieod so(lpuotmi)on (rpeapdminFg:) ((gmrLamosr) voflin(amll) (uroc/vurgy) ipnpsmamFp-lesp(iukged9.) (ruoovrFd-) 00.001271 00.1100 2200 NNoD NNDG 00002725 001100 2200 N1D5 0N1Os 00003313 001100 2200 NNDD NNDO 00004332 001100 2200 NNoO NNDD 00003274 001100 2200 NNoo NNDD 00004358 001100 2200 NNDD NNDD 0000643s 001100 2200 N1D3 0N1D3 00004428 001100 2200 NNDD NNDO 00004550 001100 2200 N1o0 0N.1D0 0601172 001100 2200 3No4 0N.3D4 0004 63 00006168 001100 2200 &% 1ND3 015 0n1D3 0004 63 00007208 001100 2200 93% N1D4 015 0N1Ds 00002216 001100 2200 NNDD NNOo 00001179 001100 2200 NNDD NNDD 00002108 001100 2200 Nnoo NNDD 00003258 001100 2200 NNOO NNDD 00003325 001100 2200 NNDD NNoD 00002228 001100 2200 NNDD NnDo 0000225 001100 2200 NNDD NnOo 0000416s 001100 2200 NNoD NNDO 0000211s 001100 22.00 NNDo NNDD 00002106 001100 22000 NNOO NNDD 00001126 001100 2200 NNDD NNDD 000011s7 001100 2200 NNDD Nnoo 000213 000133 | 63201585 XLS STUDY # 6326-159 SERUM 3847 508812iD F5F454018096..1122 F75544008870.-1122 F5544019028..1122 FF5544101746-.1122 FF5544018123..1122 FF554a019181..1122 F5544007894..1122 FSe5rau1m16B1l2k1 SSeerruumm BSpiick21 SSeerruumm SSppiki.32 `SSSeeerrruuummm SSSpopk-kc4.. SSeerruumm SSopki.? F5454100755..2244 F5454018017-.2244 F5544018068..2244 FBl5a4n0k741 .24 spBilaknek 12 ssppiikkee 32 ssppiikkee 5 sBpliaknek 63 F5B4l0ak854-24 5544018030.2264 F5F454009747..2244 F54091.24 HFCoSt5 CFoCnIeS. AMgetutaal Qsuaamnptlitey TIDiSfAB % TAE cwal Mass Mass solutio solution reading (mLior final rcvry ppm F- spiked rcvrd spiked (ppm) (0pp0m25F:)_ 0027 (g00r.1a1m00s) vol22(00mL) lug/ug) in sNNaDDmple {ug F) (uNNgODF) 00001189 001100 2200 NNDD NNDD 00001187 001100 2200 NNDo NNDD 00002216 001100 2200 NNDo N3D 00002227 001100 2200 NNoO NNDo 0000436 001100 2200 NNOD NNDD 0000642 001100 2200 NNDD NNDD 00004837 001100 2200 NNDD NNDD 00003330 001100 2200 NNDD NNDD. 00000044 6633 0000430 001100 2200 4542%% 006779 00.1155 00006779 0004 63 0050 010 20 66% 095 015 0.10 000.00000044 666333 000.0005854 000.111000 222.000 1771120%%% 111.171 000.111s55 000.111171 0000004 6633 00004585 001100 2200 6753%% 01819 001155 00.1110 00002412 001100 2200 NNDD NNDD 00001175 001100 2200 NNDD NNDD 00001168 001100 22.00 NNDD NNDD 00003113 001100 2200 NNDD NNDD 0004 63 0000232 001100 2200 43% 0N6D5 0.15 0N0D65 00000044 6633 00004447 001100 2200 6538%% 008985 001155 00008985 00000044 6633 00004572 001100 2200 6628%% 01304 001155 000.3104 0004 63 00008316 001100 2200 67% 1ND0 015 0N1D0 00002210 001100 2200 NNDO NNDD 00001169 001100 2200 NNDD NNDD 00001175 001100 2200 NNDD NNDD 0016 010 20 ND ND 00013. 1 000214 I 63291595 XLS STUD#Y 6329-159 SERUM 384. iD F5544110028:2244 FF5544000771.-2246 FF5544009953-.2244 F5F454110110-.2244 F54096-24 FF5544009708-.2244 FF5544100888--2244 FF5544008897-.2246 FF5544008902.-224 F5F454110194.-2244 serum bk 1 sseerruumm bsipkk21 F5se4r1u1m3s-p2k42 F54082-24 F5F454010181.-2244 FF5544007894--2246 F54116:24 F54113.48 F75544101812.-4488 FF5544009884--4488 F5F454017196--4488 F5F454018123--119922 F5F544101918--119922 F54079-192 F5F454101864--119922 F54113.360 F5454101812--336600 FF5544007998--336600 mFCLOo5f CoFnCcS.5 AMcattuearl Qsuaamnptliety ITIDSIAB % I n Actuala Mass Mass Ssopliutkieod so(lputpimon _riepapdminF2g) ((gmraLmosr) volfin(amlb|) (rucgv/urgy) inpspammpFl-e s(upgikFe)d (rucgurid) 00001134 001100 22.00 NNoD NNDD 00002105 010 010 20 20 No ND ND ND 00001143 001100 2200 NNDD NNDD 00001112 001100 2200 NNDD NNOD 0013 010 20 0025 010 20 no ND ND no 00001134 010 010 2.0 20 NO NO no No 0013 010 20 0014 010 20 NO ND No No 00001164 001100 2200 NNDD NNDD 0016 010 20 0015 010 20 ND ND ND ND 00001369 0.10 010 20 20 ND ND ND NO 0004 63 00007218 001100 2200 94% N1.D4 0.15 0N.1D4 0004 63 00008624 001100 2200 109% 11.36 0.15 00.1163 0085 0.10 2.0 0063 0.10 20 11 011 13 013 0064 0061 010 010 20 2.0 1132 00.1132 00005649 010 010 20 20 12 0.12 13 013 0068 010 20 0066 010 20 14 0.18 13 013 0073 010 20 0067 010 20 15 0.15 13 013 0091 0.10 20 0059 010 20 18 0.18 1.2 012 0085 010 2.0 0080 010 20 17 0.17 16 0.16 0073 010 20 0063 010 2.0 15 0.15 13 013 00006678 001100 2200 1134 001s3 00007675 001100 2200 1153 00.1153 0065 010 20 0043 010 20 13 0.13 ND 0.087 0083 0054 010 0.10 20 20 113 001113 0063 010 20 13 013 000215 rong 1 63201595XLS STUD#Y 6328-159 SERUM 384 | mFiC3o5f CFoCnYc5. AMcettuearl Qsuaanmtpiltey' TIDSFAB % Acta ual Mass Mass 0 Ssopliutkieod so(lpuotmi)on (rpepadminF:g) ((gmrLamosr] volfin(amlb) (ruegv/ruyg) ppm Fin sample spiked (ug F) revid (ug F-) 754084-360 F54116-360 0.063 0063 0.10 010 2.0 20 1T.4a 00.1144 SSeerruumm BBikk 21 00003225 001100 2200 NNOD NNOD SSeerruummSSppkk 21 00000044 6633 00007669 001100 2200 19011%% 1145 001155 00..1154 F5F454018121--002222 00004415 001100 2200 NNDD NNDD F5544101769-.002222 00004450 010 010 20 20 No ND ND ND F5F45048121.10-20828 0000438 001100 2200 NNDD NNDD F54079-028 F54116-028 0033 003 010 010 20 20 NNoO NNDD F5454009999--84 00002268 001100 22.00 NNDD NNDD FF5544009990.-2142 00008774 010 010 20 20 15 015 17 0.17 FF5544009999--41892 000034%7 001100 2200 NNOO NNOO F54099-360 F541124 0020 008 010 010 20 20 NNOO NNDD FFs5e4111122.812 000058%6 010 010 20 20 12 012 13 013 FF5544111122-:4284 00005616 001100 2200 1130 001130 Fs4112192 F54112-360 00s 0041 010 010 20 20 N12D 0N.1O2 SERUMSPIKES3-1 0004 SERUMSPIKE632 0004 63 63 0059 0079 010 010 20 20 17085%% 1162 001155 O01f2 SERUMSPIKES3-3 0004 SERUM SPIKE-126-1 0004 63 126 0074 0083 010 010 20 20 5&7%% 1175 003105 001175 SERUMSPIKE-1262 0004 SERUM SPIKE-126-3 0004 126 126 042 012 010 010 20 20 7779%% 2243 003300 002243 serum blank-1 serum blank-2 010 0040 010 010 20 20 N2o0 0N2O0 sspeirkuem63b-l1ank-3 0004 63 0co0t8s 001100 2200 7% 1N2D 015 0N1D2 spike 632 spike 63-3 0004 63 0004 63 0058 010 0092 010 20 7% 20 121% 12 18 015 012 015 018 ssppiikkee 6633.45 00000044 6633 00005553 001100 2200 7730%% 1111 001155 001111 ssppiikkee 8633-67 00000044 6633 00007528 001100 2200 975%% 1142 001155 001142 Sspeirkuem6b3i-n8k 1 0004 63 0000640s 010 010 20 78% 20 12 I) 015 012 NO SSeeruummspbiinkke2631 0004 63 0001127 010 010 20 20 153% NO 23 NO 015 023 Seumspike 632 0004 63 021 010 20 273% 41 015 041 000216 o0eL3e 1 63291595XLS STUDY # 632-159 SERUM 384 iD "`SSeerruummbBllaanknk21 ``SSeerruumm ssppiikkee 21 `Serum spike 3 `Serum spike 4 Serumspike 5 Serum spike6 Serumspike 7 Serum blank 3 Serumblank F54088-48 4 F54089-48 F54089-48 F54087-48 F54080-48 F54002-48 F54100-48 4114-48 Fs4088 dy8 F54009 dys serum spike 8 sseerruumm ssppiikkee 910 SERUMBLK 1 SERUMBLK SERUMSPK 2 1 SERUM SPK 2 SERUM SPK 3 FF5544008897-.D0YY88 FF5544008002-.00YY88 F54108.0Y8 FF5544018184.-1D5Y8 F54099-15 F54089-15 F54087-15 F54080-15 F54002-15 F54108.15 F54114.15 F54089.022 FF5544100897.-002222 F54114.022 F54089-028 F54087.028 mlof FC95 Cone. FC95 "Actual Quantity. Di Meter sample TISAB % 5 Actual Mass Mass Ssolpuitkioe.d so(lpuptmi)on (rpepadminFg) ((grmaomsr) volfin(amlL) (urge/vruyg) inpspammpFl-e spiked (ug F) rovrd (ug Fo) 0031 0021 010 010 20 20 NO NO ND ND 0004 63 0004 63 007 0068 010 010 20 20 98% 8% 1.5 13 015 0.15 015 0.13 0004 63 0004 63 0065 0.10 0060 0.10 20 8% 20 80% 13 12 015 013 015 012 0004 63 0004 63 0070 010 0069 0.10 20 93% 20 91% 14 1.4 015 0.14 015 0.14 0004 63 0068 0.10 0023 010 20 90% 20 1.4 ND 015 0.14 ND 0020 0019 010 010 20 20 ND ND ND ND 0021 0017 010 010 20 20 ND ND ND ND 0020 010 20 0017 010 20 ND NO ND ND 0016 010 20 002 010 20 ND NO ND NO 002 0018 010 010 20 20 ND ND NO ND 0004 63 0014 010 0059 010 20 20 78% NO 12 ND 015 0.12 0004 63 0004 63 0060 0060 010 010 20 79% 20 79% 12 12 015 012 015 012 0031 010 20 0016 010 20 ND ND ND ND 0004 0004 63 63 005% 010 0069 010 20 20 74% 91% 11 14 015 0.11 015 0.14 0004 63 0072 010 20 95% 0026 010 20 14 ND 015 0.14 ND 0029 010 20 0029 010 20 ND ND NO ND 003 0028 010 010 20 20 ND ND ND NO 0035 0010 010 010 20 20 ND ND ND ND 0026 010 20 002 010 20 ND NO NO ND 0021 010 20 0018 010 20 ND ND ND NO 002 0025 010 010 20 20 NO ND ND ND 0020 0019 010 010 2.0 20 ND NO ND ND 0018 010 20 0019 010 20 NO ND NO ND 0019 010 2.0 0018 010 20 ND ND ND ND 0018 010 20 ND ND 000217 eeLD 63201595 XLS STUD#Y 6329-159 SERUM 384 1 Tmo + Cone AGtial | Quantity' TORI EE ee Em iD solFuCti3o5. solFuCti9o5n spiked (ppm) rMeaadtienrg (ppm F:) s(ammLpolre (grams) TfiInaSlABe%vry vol (mk) (up/ug) ApcptFmain sample Msapisksed (ug F) rMoavsisd (ug F) FFSEtAT1040d-02208 00001270 00.1100 2200 N0D) NNDD SSEERRUUMM SSPPIIKKEE663354 00000044 6633 00003419 001100 2200 4615%% 006928 00.1155 00..01602 SSEERRUUMMSSPPIIKKEEG63376 00000044 6633 00006510 001100 2200 7818%% 1122 001155 001122 SSEERRUUMM BBLLKK 21 00003117 001100 2200 NNDD NNDD SSEERRUUMMSSPPKKE63-312 00000044 6633 00006718 001100 2200 18013%% 1162 001155 001162 FS5E4R10U5M.4S8PK633 0004 63 000m6!2 001100 2200 8% N1D2 015 0[102 F54075.48 0s 010 20 ND no 000218 ~ee13s 1% CFoinotrroel EFssiost FFssasootsss FFssisliooss ess107 FSEssimiogso/k3ng EEssssoo FFssii0i9o70 FFssttii0o2s Elsoamugke FF3sii009935 FsEssioon FFssiso0990s Fssit0 0Famnpgkg FFssiiootns7 pEsrsoy FFs3t9io9s FFssta1n0i9e S3W4m07g6ke FFssaaoonm FFssiaooonss EFsialil EFsaailis HWI 6329-159, AMDT 042095.1 ppmF-inserum h0o6u0r h0o3u04r 10h3ou6r 204h2e8ur so0iss0 o03a1 oosssc o0s3o4 oosszs ooalne oolsess ooias 0os4s oosss oosss 0o3s6 ods 0304 038 0306 Suenmtemrainrg y"NoDf"O.rioTnhsiesrsuummamnaarlyysiiswwietdhtoout diseaterpmineptherrtoeuloastievpveahlarulefswiihfi,cathhaertreefoersesit han the detetin imi 40oh4sou1or o08hu3os2ur I0o2b8eo9ur 240033ho%0ur 0oa318s 0oasms oonsss 003308 o0a4t0s8 003a6m 0oas o0d3o4 0o3dso 0022s8t 0oaa 0o25m0 04h3o%ur Sohsomu 2ohuo2ur h0o3u2r o0s3s6 oosxi4 o0m3e 00a2ms 0o2a%z ooas oodzss 00220s 0033% o0o3s o onsoo os 032 ism 032 ox Sohsosw Soheo:u Z0h3o8u t0h3o20u 4hour 1%2hour 3@Dhour S28hour 672hour 004s9s%2 00468a 00382 00220 0038% oosss 000464 00%302 00335600 o0d3o7 oocmd 0o3s0 0o%m 0o5s2o oossu oLsess 012s5 oss oss osm 001786 O0s6s 0042140 0033026 0o4as o0Sdet 005400 003%40 00430360 04h3o0ur Sohsouor Io2hiowr 24bour 4Shour 192hour 360bour S28hour 672hour 2L3200 o12m8s8 006s6 LLI0U4 L13I1S8 1Le3s6s 012%66 00981028 00696604 033s2 olomw oosmoe 1i2m2e l1masm i1s3e4 1L3o0 0Is7l%s oummee 0Do2o 1tam l1a3m 1L2i%s o12s2 osm omo 0001 2L0S0 o3s4s0 005s0 ilmaet 11376s 112s% 113980 00 07s 000219 does Fue zg ppm Fin Serum (Group ) IE zi WN i TT od < Te a 8 8g3 v - = =TieFrosos-y = = ly 16-384 Fue 1 o - c PomFeinserum(Group 5) e 1 Ez o 1] se S i e Te e Fr : 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. 000222 000142 RE: 6329-159 SERUM SAMPLES AMDT 42095.1 Date of Analysis: May 4, 8, 9, 11, 24 and May 25, 1995 Analyst: DOW @-22-43 WR Skala Qk The samples are burned in the Dohrman at 958 C using 8.18 mL of the serum. The gas is collected in 2.8 mL of 1:1 TISAB/Milli-Q water. The samples are then analyzed on a Skalar Segmented Flow Analyzer using the lon Specific Electrode (ISE) Method. TISAB buffer is added to each sample as it proceeds through the system. The sample then goes through a heated mixing coil before the potential between the ion selective electrode and the reference electrode is measured. The signal is amplified and related to the fluoride concentration. The instrument was calibrated in the ranges of 8.815 - 8.15 ppm and 8.15 - 1.58 ppm fluoride. The standard curve for the high range was plotted using the inverse logarithm option. The standard curve for the low range is linear. Rll standards and samples were then calculated by the Skalar software using these curves. All results below 0.8001 ppm appear on the raw data as #.####. A quality control standard was analyzed every 18 samples to checkfor accuracy and drift. Raw data is taken from the appropriate calibrated range of the Skalar printout and summarized on an Excel spreadsheet. The final results are adjusted for the collection volume and any subsequent dilutions. 000223 000143 =... 4.91 SUMMARYof6329-159 SERUM SAMPLES AMDT 42095.1 ans wa Skagen barroiwa:io BJ as o0sst BEFaiosnmss om BBBaaawso ooomww ooIss l o0a8 oomnL om om oom nobofw o0on%m o0am0e oolmnm ox sFBaoernmr 0m om om om ao1mm om om Dolcorosw:mrs FBEoeuann 0o3w o0nb olmm oooemn FBBinrin oomn oonm oooe oooamm BBaimo oomn oam oosn oonm EBBooonwo oooanm oaoimws ooonwn ooiama . DolGorowosmgts fBBuomoen ooounm ooonmm ooosmo ooommm BBaaom oais asn omoo oanx mFaoowon o00s5% on os em os lm om om on os om on ox om ox DuelGorowosme BHiaowwn o00w%% oo0snp 0oomm ooimmm ooonmn ooommn aooxnn on om BEBaise 05 om o@ 1s im os an om oonk oonm aoan Bmineoo0k% 0o8m oosm om aamR am aamm omm mon me 000224 ore SUMMARY of 6329-159 SERUM SAMPLES AMDT42095.1 roves oBBaasom o2lemo 1L0%6y ao0sm% o1w0 wimso 11mm oxn 110g oomm Ewes im lw in im in lk Ge Dolo mee BB[amEn rri1em 1Lmb ilSwr lIo inm inmm 0l%e ou ow BBaalne oiaan 4i%e owmm w0n% iime disn oinm ae ie Page 2 000225 000145 Hssoans sODnoersiauahreicAernstow195o0u5r0o1t0::1s8 oso Ay il ane CISKALARDATAUIWIDATASERUMOSOS00A1 2PPo ommoSaee ai0k aap0e omom U5o sSSwmaaamm:i mooswo aoomows wloon bS7o ooSSSemaaemusseasooabins deoaahn womomnm g DWhobo SSShhaaases ooimmm ooiam meoumm 5Wi oshmSae dlwm 0la0ob owwm NI uo osmeen apooaer0t 0Bonooww ooWmwm ew em os ow wx o3boo sSShhouenns aao0no 03300 ooommm ollmok tsdomom eoswmm ooomns oooisms ammmik 5nnoo SGSionee o0on5n I10 oooklm 2si6am oowms hbmeemmm ooomnw oooumn iowww u Lo0 SkTihes ao oine os 10 Gl dm ow mem ow on om i3he8 2 5 5m7 eeBWeaesnh a0a0o00 0Bonom ooaw P sSHiEe 5EE ER TH ww 0 on a {#3 a 3 Estos 002 0 ow ox fi 3 Pace Tnisoans nnFE woas 0oo0nns 0BFr omaw wons B%no osmsoaess oaoaino 000 aaonww 1oi5mm ooowmm eoomwm aaosss ooospw ooumwm no sheSmawewo aod0ne ow 00 ow am eo mem ow om ww oa& sSSooeee oooiiis oaommw aa2mmm ooommm mmmwmw ooowww ooommm aasmw B6"o sSeeannsee aopinrs wBo0 ow0okb oamm oomm mmmw oonn oonm owww o aG roTmemuw oa0wos w0o ooonnw o0osw% 8gg %oin mowaaem um w0iww om 0 on os 8 N5 D5 rRmeesswss owwoow B0o aoawwo 0oo%nw %58 Ewomasa owwown 0B0 owooon ooonuw a5 rSwiiaor owonmm nooonnoon ovake 2g@ar oumm ae w0mw o 0 oo on 2 FGEE rem a0o wFoT w ]om 3 en ww won os Paez HasANs EFsutsis wn Este oona on 21 o00o ooox 0 ow ow nao Eessoss no Ess oooo oo 00 ooww oonx 0 ow om E %noY Esos bieoonm am 0 ow om n7o EEsssoe no psn oooooo 00 ooww oomn 0 ow on 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55F50R 8 eiGimfm 150 re ooo14nnn8 ox oom o 0ommwe aos 855 Fi m re ooouonnn nnBnoo oomm eooomwmm oo g 5 mEr rem we oooinn nnnooomuonin 02 8E umSm SE one oooounnn n L0 omowo m w 2 8 Rm os No "3 [  owmen am 0a0s0 Hiss. 58 RB g 3BQ2s Pages 19350611325 outputor: susanhr rsscs RODpaetrnaotfarnFeyilAeNnaaslmysoeisw1+ 9C9I5S.K0A5.L0A8R0D7A:T20AUWIOATAISERUMSSOS08A1 :DTo ombSeamn1s do1eos oomw 55 SScwwuaewmns ooonmms o0b0io lowowww oo7O SSSauaaesss ooonnm 0oo0dm0s liooorww 70fo SSSewmaimmsss o013i3 o1omam nlwkko S S gh u k swTSUaam l1se odiieo oomm orBomsmaa proome L0oew w wow 0 ob le om am af Bo ee Ssae ooowee L0 owoowb ooAaen om am L55o sssaoariios ooaooos 00 ooonww oooemw 2 2meraomsmta aso 010s1 ot oom noon om Sa eum oo 0 aw om 2q Bon wn2 RmF5o4a1m04ma12 00a00c48s 2B 200 ooolw 0ai8nm1 paet oons oomn wwomw Es33ua3y S= ng[3d14 P 2d ses 25 Rmfseoens 3[5 aRReernn io5ao TeRmein we G5"o RRmeinn "ii mHeonw Sg$ BfB8o OTFFTutwman oe 8S5 pnHieennn 855 a rHHeennn me bF o FRe eunnn 2gHg a iGne hn 2 B 3a 6 RRreeemwni oo00usn oWoooennr wm oooiams ooommn ooowxx oofouer om oooisns ooonnn oooann oS ons e ooowra oono B0 ooow mte moeam o FFER o BwBoootewe Book eoowmm om F BoeooT m ooww Bn Booto ue ooons BF 0noott0eeTooonxa BBLooouowme on on BBnooooonooooaa w BF no owT o ]wine voz ar o@s Eosmai ow 10027 om 0 ow im no wes 000 i88 2 3RaB Pages 06010 ouput:ssn Hissoxs DOAapecrpnrbieikAarsesb:oCw9A5S0K5AL.A3R0B:A8TAUIWIDATASERUMOSC509A1 51PtTem 015s I0ar%e ow 5 SsSmhwaadmtsa ooonmms oaonows wwoowwe 5$Do SSShhmaaawadsss ooownn aoabmms odoommm nWoo SSSaammmaaer 0o13r1 oon mm wwmim gg bon hn sN wVaUn W 13 mai0n awem 8 & 6 sSaaanbwaa aooooosnr I 0Bonoo ouwa EE EE 0%Bb SSSohmmaa:m aoo0oor 0I1o aoobl o1Inbn ooommm aoommm oooiwi ooonnm amamim 2BNDo SSSnomemsa aawl00 0100 ooomlw aa1mn6 oooeomsm ooammm oobsti ooonnn bmoewe g0F nIXoO GeTNEmaatma e 0adm0 wm 0 ok oo am os os ww a IBEHS ooBeEeasna Goons 00 0o0w oine STuiiss f3 I0 Roew boi o nnoo oe on f{e26 qa Faot : nse n3NoPDso ooounn 2BFE ooonw oann 3NBoo mHeennan ooiioi NB noooomo aaammm o5 Boetm we ooine om 0 om oa 8 FsEo m nom oooaanr ou nBBoo0om0s 0oWhm oeow aawn oom omem aw Boon dh om om ow on Ds5oommme ooonnu BBBooooddne um ooommm aooeee ooonww ooonnn aaom mm ggounbPA mmetu ow iooInnn wm n Noo ooam R8 E5A mhmean ooonnn wBFooRonEm oonn 5L5 hhmeenn ooonann BBBoooooonnn ooonnn 5a5 mhDueemnna oooian BBBooodkao ooanwn a a Fema PR on on nom a og3 4G 6 mnmeew ooonnn nBBoaoonan aoawne A] on nom 3 Paez Jn oxs PonR nann 5 Fe ooosnn on 0BIo ooomw eooomwm Boom uh ales | 3nDFER oonu nnoowm ue loaded = {ag ren -sagd gs iin Wash 0 0mm 000 Hoag 6n[ nooE+ swmmsone o0oow0nn6 um on BB 2o b0 ooo o iooao mo1a 5omnooimwe ooonnn oe FnBr ooowImanom oom ib oom oan ooo gEeRBB rm nRTmit ow pi ot om nono 0 wm & u5fmF oens e BOTY we 0oool0nwwe8 om nn2L0 ooonmooowmi10 3 DTYo ooonme umIJ0Hhmmam So H H D fTe h m we oooinnn m 0nnoommweowoomn g3 1 ow 3gq ras OJ pestor sow ---- Hisexs D[ ieoftheAnalyst: 198A 5.05.11 08:16 -- 1 FR Toeme i0w aw10rg oe Po Gssawawtzs ooowmms ooammme oooowmn To ScSuuuedssss oo0lns 0oOl%te ollomow 12 SSSwwaamwds7s 0o1on% 1oo2sm3l woowm o8 HTno ousmwme i1k% atiteo wwme 88Sloow rrroiemeae aaoooorsr 0wowaow iinn @ normsas Zaina aoaosir nwoownon diisyn aBWo earFneaieies aaactces wnwoooowwo iiisne 503 o 7iBm FCweomma ue 0at0oa0s owe noo ie 2 Brauer ats 0 ow us 8SE0h7Ya She S923 5%= FrFeleneeme aoa0c0re8 0wnoonwow i1i ms 7 3E Panes misses 2a Nar] anm oooouotn wnBoooowom oonw 555 HHFoowowe ooounr BFERR oon ia 2o BOrEe we ootiae 5 ou om FPR ETE bFra oown aoonhr BB Bo0on0m iiWmn eomm aoem aonm oooonn owwa olo rmremoosnn ooonan BBooooonmom iiinmm os Go B o oFomne on ooTmuE am WER oon am 5 gS 5& mnoSmmmean ouooam Bwoann oiwm g 83a fHFioenomsm ipooHo Bnootooeobn aoien $ 5FemF)a oooonnt BBooooommoo aoinmn Go roms oa 3 ow oa PEEa6a1 F0 ETSOe0 oe0o2 uoTnm 0 we0w1 a00 om 0 Blank 1asaabd in bray no Shs sveksania tags, 2EE8 mm oooo Fnoo RETiw "33 1 Fsatz 006 20 Paoewz 12s misses o & efFesma aooonu FBBEooEooTnn aimm nBRo SeCesst aaaoot BBWooooomnne mrhoeammm aaawme [ BnoO a Sw we xatne om 2 ew am om mm nBBoo OGsEimmse eaohse 0Bo0dke dImn ooomm mmme ER on oooonnm ooaoownn moommm ooaown oaoonnn woowmm 233883 232H 3 a3 misses seen ouatisusiar Ope Dertatorniea AmneiDO9CW8OS4K5A3L4A1R3D9ATAIWIDATASERUMOSS3401 DoVo toem n us evaieo oomm CU5o osSmmnan: ooommms oaomow uoommm DDoo SSSmmeaamddass oaonnm ooonmw dwwoomwm Sh0bo SSSawammmaaayr o0orpc aoiama dwome gS BoN onnsame o1n odamww oomm W B baw on oIromn 0BBooowann ooVnwe em aw nFbi oowesn ooommm BWBooaowwn iimmwo aoomme aaaeme Bo as om Ik Dn oom an g Sn EB1o ShKeEsS bm me oo0mn08. om 2BF0O ol00 1da6 0a04 @a0e0 25B 0%5 aa iwnn 0 aw am pant ooahmn aoonsn uhlmmn oann ooawn hore gey 0o1ns 0o1e wnomk EFWfReAiC S}pHi3At rss 53Nerhnoonwass ooonnn FBnoEooEenT aonn %3No THHooonnwss oooinn LLNoo ooooek oooaonn 3LBo rThoimsns ooonnn BnBoo ooomonaonw 5 meOTmEon edeoonn mm 0 on ou oh65mm Taew ooonnm BBFO ootdeh dVSa domm AGanw ggSkn h mamn oioonn F Booo deTaWnb om aw a PK632 008 20 010 155 004 6300 N 5D i"o T SrPKo63n3 we 0oot0inn8 om 2H0 om010 e16e8 oom 600 N55o Hbmoaosnonnn ooonnu FInR toew a on SERo 835 abomwaonon oioonu on BNBoooooann ooannm Boom on 2 g Ea G E ranosn oooann noooooooo oon Paez Aaosn oown ol+ww es oon om 01s 015 101% 0s on nk nes aa om an mon oownie om a@ bFraaonmoonnn oooann aoTorrhnwo oooaan 3n BHioonseoonn oooannn Sg OnTsNEhim we ootines om 2g 85 nn5on SmmmPKaaa61ssl4 oio0o0nn4 0F BOT sea we ooWnn e FI on BB0ooooommw ooowmn BBBoooooonnn ooonnm F BBoooonHoonn nooo 0 ow BBB20oooodtk0eh10 BER ook ou 0WiI7nn%odo0dm0mm04oaa6G%0en0 on o0ao1onsnme0oono0nk8 osooommw% 2283 a ras 995-05-04 16:44 OutPut of : 950504A1 AMDT 42048S.| eee oftware : version - 6.1 1990,93 2-15 Sean BESTCOPY AVAILAeBxLE 1s ate of the Analysis : 1995-05-04 08:18 nalysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950504A1 `alibration order = Inverse Logarithm lope 1S = R.EREEE [== ] esult = 10Ll s C1 = corrected valu of the concentration 1 s = Slope of the electrode 1= 0.00063 0 = ~1.24458 ampler iluter Type Sample Time : SA1000 : 50 sec. needle Height : 70 mm. needle Height : 80 mm dilution Factor : 10 000244 000164 an s95-05-08 36:88 -- luoride 1.5 SnpPeaettoIheennTumabEeerrr + p3huies |8 Resample EHP EEr eInoNa | E15s diG output : No :0 ---- 33s32 mmsgTahmnmdtaar:d:: diIgamnoeere 2s36 omsooTamomndammr:d 0oso.o1nn5m0 SSS5h1p0iemirseTeasnTdeoaorvrde,rds: ek1i.5i00t en au E IEm,aRn T trigger Limit : 1800 Sec BESTCoAvpa pypg soride fPoriamsmeracion eNd ignore Measure window 2| 2: 120 : 75 Sec A] = formula : BpMSraaEendTlaonwuummerr i0 es Bam, diLute ik+ No BHieTmre.s: dil Threshold : 4095 000245 000165 23 995-05-04 16:44 output of : s1 standard : s2 sTandard : s3 sTandard : s4 sTandard : 0.015 0.030 0.060 0.090 ss56 sstTaannddaarrdd :: 00..115200 $275 SstTaannddaarrdd :: IIggnnoorree 5S9lo SsTTaannddaarrdd : IIggnnoorree Doridmeernsi: on2 : PPM sttrairgtgerValLuiemit :: 1508000 SDUec PSetaAkrtshiagpneore :: 6P0ointeSdec MeNeaasureigwnoirnedow :: 71520 S%ec RFielgteenreration :: NNoo ofourmtuplaut: c#4.:#=#c#3# 950504A1 4%,py (/7A 000246 = 000166 24 995-05-04 16:44 `os Typ `i:w "a ss21 i $3 s4 s$65 .0 $s87 32 ss910 s1wa 75 ouou s8 uu 1 ou 32 wuu 5iuu 75 oaw s8 uu o1uu 32 ouu iSouu 76 ouu 9saow o1ua I2 uu i5 uou 75 ouu 89 uu oaow 2u Iu Ident InitialTraWcaesrh Darisfht SSttaannddaarrdd 21 Standard 3 Standard 4 SSttaannddaarrdd 5 SSttaannddaarrdd 87 sSttanadnadradrd 109 DWriafsth BBllkk 21 SSppkk 6633--21 SSppkk 9944..55--21 SSppkk 112266--21 SSppkk 112266--43 DWraisfht FF5344007851--44 FF5544110057--44 F540B7l4k--14 Spk B6l3k--12 Ssppkk 6633--32 Dwriafsth SSppkk 112266--21 SSppkk 112266--43 SSppkk 112266--65 SFpk54018256--47 FF5344100840--44 Dwraisfht F54077-4 F54083-4 Output of Fluoride 1.5 PH Ch Result F Time 33 10.l0a5z7 21625 33 01..045677 532807 33 00..007635 971450 3 0.090 1087 3 0.109 1263 33 00..115332 11641316 33 00..620838 11976817 33 11..427246 22133173 33 01..045772 22742897 33 001.007707 32081338 33 00..009985 33316848 33 00..008908 33753184 33 00..111721 43086328 33 00..114492 44421348 33 01..045275 44755868 33 00..007793 54191338 33 00..006663 55426951 33 00..802780 55861451 33 00..140002 65196859 33 00..111131 66531359 33 01..045678 66961819 33 00..114476 77201450 33 00..115414 77536960 33 00..115378 77971460 33 00..018239 88206952 33 00..007722 88642432 33 01..045677 98073320 3 0.071 9138 3 0.066 9314 : 950504A1 Fluoride L em Ch Result F 44 1n.a07a5n3 44 #1l.a0e7n7s5 44 00..00310540 4 0.0594 4 0.0891 44 00..11423201 44 00..52283625 44 10..09803810 44 H1..#0#8a1#0 44 00..00328452 44 00..00763866 44 00..00473332 44 00..01963978 44 00..11433413 4 4 1.0452 K.an#d 44 00..00340260 44 00..00212792 44 00..60620585 44 00..03776878 4 4 0.0929 0.0963 44 #1..#0#745%0 44 00..11431977 44 00..11436678 44 00..11523932 44 00..01419729 44 00..00229718 44 #1..e0#74725 4 0.0265 4 0.0163 Page 1 of 2 Time 00 00 00 0 0 00 00 00 00 00 00 00 00 00 0 00 00 o0 0 00 00 00 0 00 00 00 00 0 0 0000247 2 3 -2t ==] = 3 = . 000167 25 995-05-04 16:44 Output of: 950504A1 Page 2 of 2 Fluoride 1.5 Fluoride L Pe. PEM os Typ Ident Ch Result F Time s4 uou F5544100904--44 33 00..006677 99468685 TEI u 540B9l1k-41 33 00..008697 1090814s2 ssuu SBelkk 12 33 00..100634 1100139617 oiuu F540S9p7k-42 33 00..016075 1100751473 5zaow DWriafsth 33 01..045376 1110086951 5iuou FF5S44019022--44 33 00..006647 1111421491 75 ouou FF5544100860--44 33 00..007752 1111756991 8 ux P54089--+ 3 0.067 11942 03 uu o FF554477008878--44 33 00..006770 1122219116 21 uus FSF4S73101906--44 33 00..006641 1122644622 i3 aou F540D9r0i-f4t 33 01..046545 1122989128 55 ouw F5407w8a-s4h 33 00..006577 1133314529 57 uu FF5544019051--44 33 00..006631 1133559126 osuu FF5544007913--44 33 00..006638 1143806482 21au FF5544111068--44 33 00..103652 1144329133 i3 uuw FF5544009884--44 33 00..100981 1144576484 55 au 540D7r9i-f4t 33 10..417582 1154093139 57 uow FS41W1a3-s4h 33 00..102547 1155432342 oiuu FFS544101812--44 33 00..10391 1155671995 izuu FF5S44101746--44 33 00..008802 1156916475 i3a F541D0r9i-f4t 33 10..406685 1166341393 t5 orww RunOut WWaasshh 33 00..005577 1166695608 Ch Result F Time %, 44 00..00119861 44 00..00212e01 44 00..00810300 44 00..00280305 44 #1..#0#5#3#7 44 00..00114912 44 00..00324807 4 0.0181 44 00..00220408 44 00..00102634 44 01..00163881 44 0#..0#1%9#4# 44 00..00017260 44 00..00211138 44 00..01130027 44 00..00682849 44 01..10487517 44 0F..141#1#4k 44 00..10271002 44 00..00446301 44 01..00176800 44 ##..######## 00 % 0 22, 00 00 00 00 00 0 5 0[EESTI 00 00 00 00 00 00 00 00 00 00 00 00 o0 000248 000165 20 ------------------------ Raw data of 95050481 : Flucride 1.5 ----m 4085 Tr a 1 Iii} I it | +] ill 34 i! i}l Ji | "+ | ;i1 ilI 4% il | li i i i il i i ig It i i. ii H HH o i| I i 1i 50 | i I. i iI iI i4 i | i i | i Ii tH iH ii i iHi i| II i i J}| H1Ei |i STqtyenes ius |} 1i | L | s PH LA LAERS ARh S (E Ea R AAERE aa inhe I i a AA a A R VU LweA YY L AeAn] se wAAN | 0. EN NE Tine | ' = SSTEXIt | Fisheln | Cril-PeEdit peaks | 000249 000172 a -------------------- Raw datz of S50S04A! @ Fluoride 4.5 -- feattche a ei EHD Use SRETAEIEDe to EER or rr 4088 - i wir "{ iiii! 1 Iy;1 i iiii i i i # it i Ji i; i iI| ii7ll i!il fIi| 3h I he Ti hIi i i|Ii i [a HIIn l Hi a| J I!ili I| i1 i1]| ii|1 . It i Sa ih Ii|i Iii | iy ! 1|| | || 1 | Ii | i i 0 | I | a An Ad Aa ALl Ge eAe nRAeG MeAE g@eAmaloRwAhTeTme| 8780 Tine 13780 | se=Exit | Fi=Help | Crtl-P=Edit peaks ! 000250 OTT -------------- Rau data of S50504A Fluoride 4.5 -- 1085 1 Pathos 3:2 Vie KRICA. atoss MRERUNNEDott: TEtx ierra | El 4 : iq1| i4 11| iii 1 i To!}! iti i Poi | Tl i i 1| ii"| | I 1 i ii! | 1H ;1 i i i oo | !i ||i i iii? iIt i Il I1 li IIHh l w) Hef ee fe # | | i1 | i i| i ! ifI| I} [1 | I ii |[ Lo i ! | I | Iii | i iH| iI| il I. Il | 1i iHii ' n' a ol oE n MTAR h Hanil W haee| || fl ATR WUT WR ee i 04 Po 12435 | ! ! ine | ! 17435 | sc=Exit | Flshelp | Crti-PsEdit peaks | . 000251 000174 995-05-04 16:48 Output of : 95050481 seestoe - BESTCOPY AVAILABLE oftware : version 6.1 1990,93 ate of the Analysis : 1995-05-04 13:07 A0Cnwo"h4a2l8a9s5 RAu3S, nalysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\95050431 alluiobrirdaetio1n.5order = Inverse Logarithm `1ope BEER [x=-=el] esult = 10l 5 xC1 == ccoorrrreecctteedd vvaalluuee ooff tthhee scaomnpcleentration 1 s = Slope of the electrode 21== -00..0000006020 0 = -1.24651 alluiobrirdaetioILn order = 2 `orrelation : r = 0.99962 es=ual 2*tx3 + a *xl +a0 21== 00..0000000200 0= 0.00460 ampler TNyupmeber :i S1A1000 Wsaasmphle TTiimmee :: 15200 sseecc. Air Time : 1 sec. Tsapkeeciaulp :: sNionngele needle Height : 70 mm iluter ndeieldulteionHeiFgahcttor :: 8100 mm Redsilaumtpiloen Volume :i 21.5 ml. Dilution runs : 1 URseeprrocfeisle :: No Et 000252 000175 - 995-05-04 16:48 Output of : luoride 1.5 SPaitghnalnumtbyepre :: De3bubbled DescysotleomrNumber :: Y0es Rdietsuatmeple : Ntoo da4iilG oTuhtrpeusthold i: 04095 Window event : Off ss21 ssTTaannddaarrdd :: IIggnnoorree 5s3i ssTTaannddaarrdd :: IIggnnoorree $$65 ssTTaannddaarrdd :: Ign0or.e150 s$78 ssTTaannddaarrdd :: 00..630000 5$910 ssTTaannddaarrdd :: 11..520000 ODridmeernsi: onInv: erPsPeM Logarithm tstrairgtgerValLuiemit :: 1580000 SDeUc sPteAarktshiagpneore :: 6P0ointeSdec MeeNadsureigwnionredow :: 71520 S%ec RFielgteenreration :: NNoo ofuotrpmuutla :: ##.### `luoride L SPiagtnhalnumtbyepre :: De0bubbled Descysotleomr Number :: 0No RdieLsuatmeple :: NNoo ddiilG oTuhtrpeusthold :: 04095 Window event : Off 95050481 BEST COPY AVAILABLE 000253 000176 995-05-04 16:48 OutPut of : sl sTandard : $2 sTandard : 00..003105 $3 sTandard : s4 sTandard : 0.060 0.090 s5 sTandard : 0.120 s6 sTandard : 0.150 s7 sTandard : Ignore s8 sTandard : Ignore 59 sTandard : Ignore 510 sTandard : Ignore order : 2 Dimension : PPM start Value : 500 DU PteraikggesrhapLeimit :: 1P8o0i0nteSdec start ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula : c4:=c3 output i #.@### 950504B1 000254 000177 - 995-05-04 16:48 OutPut of : 950504B1 `0s Typ itiw I s1 s2 s3 |0 Ss3347 3iD 33l55o 4P5 Hi ud [sHaH u HHii 3i Saoau aHHii u 30HFai au qua 5a6 a4u3% 3 0DSaau i SIaa 5HSuEou 0FPPa uad eonFluoride 1.5 Ident Ch Result F Time InitialTrBaWrciaetsrth 3 33 101...044576737 65 2a1s2s Vash 3 0.087 620 Standard 1 3 0.065 740 Standard 2 3 0.073 910 Standard 3 3 0.089 1089 SSSGtttiaiannnndadaaarairdraa7dc4 3333 o0o0..ls170ess5i 1TiI2sh76ie34so0 sSSttitanandndadasarara18dso 333 0i1.l760ss5 i2hs1ee3d FFsli0Doa7rs5isofshbt 3333 1000....40004518873 23234703081)083 FFFS5ii40l1o07s7iies0 333 00o..l00c58e15s i3sie1l FFiliocessss 33 0o.l0o7r3 naszi BBBSLirKkai--e1zt 3333 000L...a100168z388 ad440ss26ss17 FseiSoWRia-sSsh 333 000.l00073s31s 4s513a10i2 FFFF5333444i001o700e710a--.s800 3>>3 0000..000088765527 5353i680a411123 7F5344005971-66 >3 00..006810 s3i33d7a FF5i4iD1lar0oi2ssf-hst8 3333 0100...00035653557 6ac63a033s39o4 F5FF5is94li00o079io15c3-0ec8 3333 o000l...o000e686r572 7777039349180354 FF73ii4id0or9sa05s6 333 o00.l00e36ss3 s77o39d41s86 BT3ih4oi0sl8ai7co8ss 33> 000..100076038 8soe4sa4re1s FFsiiD0larneissfoshtb 33> 3 010...400857176 oles o8897i39r80 eas eonFluoride L Ch Result F Time 4 s.asss 44 0o.l8s003e7e 0 00 4 9.9m 0 4 0.0140 0 4 0.0311 0 4 0.0615 0 444 0000.il20ii8a9d70e%7es 000 444 o00.u47e40ns% o00 4444 0o04..i7o0e91s5m8s10s 0o0 444 o00l..000311m38s4 oo 44 00l.0m0m31 00 4444 o000l..0%01s209e10%s1 [00J 0 444 b09..00ar5n06sa8s 50o 444400..00.00l0530230397872 [o04 0 440.00.003071 00 4444 060#..li09oe109e409r6s7 00o0 44440.000.00.0330004083s81 50oo 444 030ll9oo8i%9oesrs o00 444 o00l..0002300n07 0 5 444 60s...68a41s70e63e 503 19.0318 Page 1 of 2 %, 2D, % % 000255 000178 995-05-04 16:48 Output of : 95050481 Fluoride 1.5 Fluoride . Pry BE os Typ Ident Ch Result F Time Ch Result F Time s5HE uou F355444100890025-.888 333 000...000777554 99468964171 444 000...000343633433 000 7suu FF5544101746--88 33 00..008777 1100011553 44 00..00548052 00 55 uwu FF5544018121--88 33 00..100853 1100356473 44 00..00857973 00 Lzua F541D1r3i-f8t 33 01..059142 1100879117 44 00..80274070 00 3FE ow F540W7a9-s8h 33 00..005973 1111029430 44 40..40468841 00 iouu FF5544008948--38 33 00..009959 1111451961 44 00..00771982 00 7sdou F54D1r1i6f-t8 33 01..252182 1111796461 44 00..28223465 00 5 wrw Runout WWaasshh 33 00..005577 1122049176 44 ##..####e### 00 Page 2 of 2 %, 22,Z,e 2, 000256 000179 -------------------- Calibration curve of S5050481 : Fluoride L -- 2.1816088 1 ! [I wed >= J8 1| / / byi === - 1| ~A i = 2 | / bi E / i 5 3 A i / || 31 J I. Hi / I 3d + ! | / i - i1 p' ] ~ 1 A I I 6.004503 3 a Order : 2 Heasured i{1 i by i r 0.22862 i Sno i 000257 000150 ------------ Calibration curve of 95050481 Fluoride 1.5 ------ :}|1 5SiH: H| 1 863943 1 Ad i / | 4 A Ll Z| / | H / [I 5gi /J i | / } 4 / bl 0565535 | + H i / | i Ve id i rd 1 ~~ | 3 oo ot 1| A -- = i| 0 Measurad 4035 | Order Inverse Logarithm 3 63e0 000258 000181 29 Rau data of 95050481 Fluoride 4.8 -- FEET 5 0851 SY Td ONY waar IN0E2 T> e! wPo 1 i bo Ph sol Po Tha Fil ! Phbod [hiti i | [PRihR li ii i | |i | 1PJ iii bl Hi ' |iiiafii to ho Po od 2 LoTi HpHiii nho| on iH it f Hi h hi o | I il H } I bio ol Ll FH {i ! 4RHi fWiii no oo Pi aii I $i Hn TH ha sHo a P{oi i wF AA E e. e a = A phi E A CEES YUE FERRY So finial In AASUYUNN eRa VA IAE AN W, N E hA i el4| 04 | Tine snc|o SSwExil i FisHelp | Crti-PeEdit pezks | 000259 , 000182 -------------------- Raw data of 95050461 : Fluoride 1.5 -------------- E Gi P BIRT TeoRR iTsass | wes J HE I i 7 Byi |i 1| i i y i i it | I 1I iti fid ii iif iIibBi l 4 | i I H I i I i! i i I Pial Iitl Ii! I I ili i i! 1 : i i 1 | I I i |! i | 1 l i I 41 1 i 1 i I I i| Iil HHh 1 iIl ae h3 "1To { ;! 1De jAAie TT2emiien"e es HfEeRnES 2UE5Ara | 0 || SVBAANA] SUVARI VL STUY\ il 4580 Tine E50 sa=Exit | Fi=Help | Crti-P=Edit peaks | 000260 000183 ------------------------ Raw data of 95050481 : Fluoride 1m .5 eee | | 1085 7 [3d ? | i5 J i i | 8 | ! i { i i ! | i | 1 i I | = | = i = 3| l I| I Ii !i: 4 1| iilliit Iil|! I||| il il I! i 1 ii Hil id; 1 | I i 1|| |tH allI" I il hy i ! ii | | > a > L% wl weSduiit iHl ! LT wn" py aba APAU RRAAY MAUAR LPRUweh URUi 4Vag i || 0| sess ' Time 13335 SeeExi|t Fiehalp Orti-PeEdit packs | 000261 000184 995-05-08 12:46 Output of : 950508A1 p aTw "4h2a0l9a5s0 soefrtawtaorre : version 6.o1 wc1990,93 BEST COPY AVAILABLE(329-159 Ser ate of the Analysis : 1995-05-08 07:30 nalysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950508A1 alluiobrirdaetio1n.5order = Inverse Logarithm slope is = goereer x - cl x = corrected value of the esult = 10l 5 s = Slope of the electrode d2 == 00..0000000606 = -1.26708 alluiobrirdaetioLn order = 2 `orrelation : r = 0.99873 es=ual 2*tx* +a *xl +a0 12 == -00..0000000201 0 = -0l00833 ampler NTyupmeber WSaasmhple TTiimmee TAiarke upTime nsepeedcilael Height +taSA1000 :: 51020 sseecc. :: S1inglseec. :: 7N0onemm. iluter ndeieldulteionHeiFgahcttor :: 8100 mm Rdeislaumtpiloen Volume t: 12.5 ml Dilution runs : 1 RUesperrocfeisle:: No TT sample 000262 .. 00018! 995-05-08 12:46 OutPut of : 950508A1 luoride 1.5 SPiagtnhalnumtbyeprs ; De3bubbled DesycsotleonrNumber 0Yes RdeaiisliauntTpehlreasnola : ; YNieoos W4iinGdoowutevpenutt:| o0ff BEstogpy Ay IE luoride L s1 sTandard : Ignore s2 sTandard : Ignore 5Ss5i3 ssstTTaaannndddaaarrrddd ::: IIIgggnnnooorrreee 76 sstTaannddaarrdd ii 00..310500 So sTandard i 0.800 OSs9lroderss:TtaanIndndavreadrrsde:: Loga11..r52i00t00hm SDtiamretnsivoanlu:s PP i 500 DU PferaikgoesrhapLeimit i| P1o8i00nteSdec MNSetdaasrtureiiggwnnioonrreedow :i 716052.0 8SSeecc FRielgteenreration |i Nvoo ofuotrpmuutl.a i #a.enn Path number : 0 DeSSyicsgotnleaonlrNtuympeber f: 0e + Debubbled adRiiellsuatmTephlreeshold |i: 4NNoo035 Waiinsdoowutpeuvtent |i O0ff 000263 000156 995-05-08 12:46 OutPut of : 950508A1 s1 sTandard : 0.015 s2 sTandard : 0.030 Ss3i sSTtaannddaarrdd : 00..006500 sS$7e5 sSStTTaaannndddaaarrrddd :| Igno00t..e112300 s8 sTandard : Ignore s9 sTandard : Ignore O$D1ir0mdeenrss:iToann3da: rdPEM: Ignore SttrairgtgervalLuienit i| 5108000 DSUec Peak shape i Pointed StArt ignore : 60 Sec FeiHNeldatseurriegnwoirnedow :I: v17e250 3Sec Regeneration i No ofuotrpmuutla :i cv4e:=sct3 000264 000157 995-05-08 12:46 OutPut of : 950508A1 JFluoride 1.5 oonFluoride L `os iwTyp IIdmeinvtiel vas 3Ch 0.08 Result F ss Time 4Ch sams Result F 0 Time :a TrDaarciefert ]33 o11..e4446m76 a2318m04 d44 e00..n7733n9230 0 0 GGs1ooooasOStmmnanEEndnnaary1d gl3 ao0e t.0ae63 maa7me3ee4 i1d4 00oo.001an3m9 80 PoD53Tso5oosnGGStmamaenemadmaiirsid:ls5 loo3 odoo0o.o1oms2n4 oLli14nkmm3s6 diso4 ios0o.om1in1ae5sn6 0300 $H3i es10 cE loCbiEa standard 10 3 1.464 2309 EisLk 4 0.7386 0 T7S:75 uY:3uuw mmemEBBSillokkEen21llE}33dooSS00..oiC00mo67nIn31 2835 mBmmeooEjsoocEseem 3010 4 0.0298 4 0.0128 0 0 8 DI$Suua HEEnfhoEEaiE oog o8oe0tNm GMeiinyt dsdisoooooohwmse od 00 ID3YUduy QeeEeaeGtnaEBnor33Gooodsiae:nns gUiHmnEEe ootiafenooaunNmm 00 SPDuUu Rm ERRdEESoeoemsm uEmyke dofoodsnoee 0od 3u PJD uu YEmeE NmNos 4 u F54100-12 3 F54091-12 3 0.071 Nfoeoogmp 0.078 5813 madaue5991 D3:ihUs3 OeBBSIp eEr)I NReEm. mSLRuO DD$PU4uuOEEE BimNI nENh eSoSoooeenmmdaaa ooaHapmznm D6$u0u mBeinad F54096-12 3 oo0.ho0e5gm8 ag80mi88 P39'U4:u eF5o40t8S8Et-1t2lL33bL O0E.eR075 8Gan61men1 Iu F54089-12 2 0.072 9315 4 0.0309 [4 EstdoaSnnosimo0 4 0.0448 0 afolsfeoUtmanA o ffirnrmiammn oO ds4 0o.0oo02m1mo00 dlo4 i0oo.n03ank97 080 4 0.0326 0 Page 1 of 2 2, 22.2, Zz 00018W5G 000265 995-05-08 12:45 outpuc of : S50508A1 eonFluoride 1.5 `os Typ Ident Ch Result F Time PSSUuUEEERslioimTariszO33 so0l.o0gs7s2 asseemems s7 uou FF5544111049--1122 33 00..007742 1100011973 9D3L4au REP54eN07DiT6r-isE1f2r 3333 c0Lcl.llo0aos7ine3s 1lGf0ooo3ad6sss7o Su Wash 3 0l0%s iloss i74D0%Uu BEEFl5ioi4oeo1ns1sie1iiu-ia1ds2s 3333 coo0l.loo0as9m:e2 iii1i1iiz2se4sss2 Su Faller 3 alos ise 9 fodwaw unoutDwwraaissfhht 333 001ll.0o5ss1ii5 1i122s91i91s16 poFluoride L Ch Result F Time s44o0ol.o0ez3s2me0 050 4400..00336222 00 44i4oo00l.l.0o37i4i01 oo50 1 nears o 4144oo00l.oo00es7ms0)s1 o55[J 1 olosss 0 114 30n.ee7r5r9ss7 :0 rage 2 of 2 % 22 Zz = 000266 000184 Calibration curve of 9505091: Fluoride L ------ SETA 3.438500 5 TWO EENTEE | on - ;1 g 1{ SE / == | p/ |BBE g|l 4' ] i i 0.003325 0 Ocdar 2 _-- "7 Hessurad iLl i i | | i ro: p.geer2Tem 000267 00190 o TO = ------------ Czlibration curve of S5IE08A1 Flueridz 1.5 -------------------- SEEREs rE Ie REelee g .; 3y =~ tl / 31 / Pt i / IB / i 3.0540784 eT ) Ordar | | teasurad Inverse Logariths i 2085 s 208 000268 000191 ------------------------ fav dats of 3COSCERY Fluoride 1.5 --0m8m8 ------ TEREST RTT arta Beene 5 wes 1 T*e LbCHoda Ld I F Polill Pl RPEoiL I HOhH Bl| FIi i Hi 1H iy i ri RN MNHiE) Til iPiLoioan d I FInh wiilha VFinh Fi H| i of FH IRIE s adH inT t 8=2 *C | B== CB i fi(bIod.1 bo ilbobodi iloi i ol 1 i| | | !| IRAE 2 Pi Qa a LEPIas Cnh r e fha aea LAoE iFP FAVAA VATRVU RVRMY ET hw aa UPA T Bde| Ly Sy 0! To tive croo GsatExit | FleHelp | Drti-PeEdit pesks | TT 000269 000192 wis i: i 1:| i iSoiul i Ji hi Rau dats of SSE0E! Fluoride s5 -- 5| i | i| ! ilIi i fi ;iit|i | = = T 0| B I]== ||' | iE |i| t| i ii| ii iIi i | ii ii| | i i iil | ! i J pha" an, i{ | iLo i i Patt Te Ta | ow wef Tee | oD EAN] leo wwe ify VY 60 '' Tine ! san | SoeExit | Fishelo | Cril-PeEdit peaks TT 000270 000193 wes A TR SI Se ---------------------- Rau dat: of JELSEEAY ry Ae EES Fluoride 1.5 ------------ i " i wr | 1i {| i1 i ii -| i|; itii || i i i i 1 | I ER i" | i ti l 34 il It ! il i w= | "3 | = os ii -=< l= =BE |LB | 2 Hlii I | | 1h i i i i ]i i| iit i I Hodes! +4pops Poet VP LAAhan { PenAn UUAAUR YAVS V TNae, sasn Trike ' GosExit |Fidhelp | Crtl-PeEdit peaks | il | i | ibi hi Tan ) 000271 00019 1995-05-09 15:09 OutPut of : 950509A1 erator ow joftwar: e version 6.1 1990,93 Jate of the Analysis : 1995-05-03 09:48 BESTCOPY aviapy \nalysis File Name + C:\SKALAR\DATA\HWIDATA\SERUM\950509A1 LL "halas 159 Sl AMDT 42045.) ea `luoride 1.5 `alibration order = Inverse Logarithm s1ope fs = assenr result = 10r{ 0x s=1 -clq Xc5xli === ccSlooorrprreeeccttofeedd tvvheaalluueeeleoocfftrottdhheee scaomnpcleentration 1 1d02 === --010..002004003602109 "`lauloirbirdaetioIn order = 2 Sorrelation : r = 0.99974 tesult = a2 * x* +al * x +a0 2== 00..0000002000 = -0.00554 sampler THyuomeber D1$A1000 WSaasmphle TTiimmee :: 51020 sseecc. ATarke upTime : 1Singlseec. snPeeecdilaelHeight :: N7o0nemm. iluter dnieleudlteionHeiFgahcttor :: 8100 mm Redislaumtpiloen Volume : 21.5 ml Dilution runs 11 URseeprrocfeisle : lo TXT 000272 000197 995-03-09 15:09 Output of : luoride 1.5 DPSaeitcghonatlonrumtbyeers :: vDae3sbubbied aiRSyetssutatememslewumber :iDN0oe Wi4iiG1ndoTowhurtepssavhteonltd:|i o04f0f95 uoride L S35si152 SSSstTrTaaaannnnaddaaaaarrrradda |::: IIIIggognnnnoooorrrreeee 2$657 SSSTrraaannnadaaasrrrada ii; Iono00r..e310300 5SS5olo SSCrmTaaamnnaadaarraaardi| 110...536000000 DOsritdmaerertn's:iVoanlIunev1 erPsPe |: Lo50g0aritDuen SPtereaihksreesrhiaapnLeoirmeit ||| 6P108o0i0nteSsdeecc FRiFeNoeldsgtseeunrreeriagtwnioironendow |i;: No71o520 3Sec ofuotrpmuutla |: anaes Spiaotnhalnutmyepre |: De0bubbled DRdSeieyctssouatltmeosomrlsNumber || NY0Nooo WaaiiislndooTwuhtrpveusethnotld |1| 0o4f0f9 950509A1 8StegPryVi 000273 000196 995-05-09 15:09 OutPut of : $s21 SsrTaannadaarrad 00..001350 Ssi3 SsTTaanndaaarrda i: 00..005600 55s765 sSsTtTaaannndddaaarrrddd : Igno00r..e113200 Se sTandard | Ignore OSs91r0derSs:TTaann2ddaarrda :: IIggnnoorree DSitmareensivoalnue: PPM : 500 DU PferaikesrhapLeimit i| P18o0i0nteSdec eSNtdart iiggnnoorree :| 1602.0 SSeecc MFeialsteurre window f| o75 & Regeneration : No ofuotrpmuutl.a |: #c4.:rie=ece3 950509A1 000274 0001Y 395-05-09 15:09 OutPut of -: 950509A1 Fluoride 1.5 Fluoride pon pon os Typ Ident Ch Result F Time Ch Result Time iw InitialTrwaacesrh 33 01..045774 20655 44 0r.7n917 00 a" Si standaDWrriadfstn1 33 3 001.1.005580752 367823464 444 0H0..o80w013s41e1 00 0 S5z3 SSttaannddaarrdd 23 33 00.1007848 1098120 44 00..00331888 0 S35i SSttaanndaaarrda 45 33 00.112087 11248305 44 00..01310983 0 0 S5e7 SSctaannadaraar7ds 33 00..125330 11571807 44 00..12570238 00 Ls Standar8d 3 0.606 1960 40.4494 0 23Hi sS910 stSatnadnardDarairf19 dt0 333 111...245364388 222134318504 444000...786829928243 000 sSUw serumbwlaksln 33 00..005870 22782369 440.H0e4s5s1 00 7u Serum blk 2 3 0.073 3011 4 0.0312 0 8OSuuu SsSeeerrruumummsspspkpk1 kz3 333 000...000888051 333313386791 44400.0..000445338764 [04 Lou serumspk 4 3 0.051 3712 40.0650 0 2u Serum spk 5 3 0.115 3887 4 0.1016 0 3iSouuu SSseeerrruuummmsssppp kkk6 e7 333 000...00095950 444024631283 44400.0..000877312338 00 sTuwa 5410DW5rai2fs4th 33 3 010...030508785 444589813733 44400.p.8a00s36e40s1 0 souu r534s007851--2244 33 00..007608 53121312 4400..00223062 o0 1zua F3344110047--2244 33 00..003750 53643854 440.0.00213563 00 Siuu r7a3s10087s4--2244 33 00..006775 33891944 4400..00137552 00 HTiEuu FF533444001890530---222444 333 000...00033555A 5 666135630258 444 ###...##wasarknsse 00 ssaw DWriafsth 33 10..045772 08962378 440.# 7t9e0e5s 00 o1Zuau BBSllokk k112 33 3 000...000787358 777023319801 444000...000318130072 0 0 EFE SSookk 23 33 00..008875 77576353 4400..00585738 0 57aua SSSppekk k645 333 000...000899832 785902156213 444 000...000566975248 000 0Esa ua 5408BD5ri-ik2f4t3 333 001l..005760858 888157318555 444 000l..o08d10i55z50 00 1zSuuow F3siil00003W--a22s4h4 333 000...000588786 939013241823 444 0#0...0p02a10n58s3 00o Page 1 of 2 2 7 2 ~Z Zz 9s 00019 000275 < 395-05-09 15:09 om mrs ons Wy OutPut of ": 950509A1 Fluoride 1.5 Fluoride 1 pon on Page 2 of 2 "ny, 3 ou 3109424 3 0.085 9863 4 0.0133 672 aua F35344i101090182--222444 333 000...000688553 11009018184282 444 000...000110339930 000 soHE uu 77F3S35411000579317-222444 33 3 000...000786044 111000357641406 4440.00..000121042621 0 HFE I DWraisfth 33 01..045973 1101803908 44 0#..7#3s9s7% FE 5409324 3 0.088 11236 4 0.020 iJouu FF5344110110--2244 33 00..006876 1111543104 44 00..00117511 0o 7zsuuu FF53341i000937608---222444 333 000...000687441 111112791741233 444 000...000152116701 o00 ou 3108924 3 0.070 12282 4 0.0242 1ZS1auu SIE FFF553444100008657---222444 3333 000S...E845037N554 I 111222846385005 444D00.0O..345833N344678 p000eriedad HiuH F54080a-s2h4 33 00.008h9 n13a33y5 44 0a.08e06 o f SEHR 7 ou Fss092-24 3 0.084 13517 4 0.0310 8Souuu FF5s4i1101B94l--k22144 333 000...000776748 111343068498233 444000...000323208249 [00J HHEE BSlekk z1 33 00..026055 11l4i3z9i2s 44 00..00184753 00 HSE u Fs411S3o-k2:2 33 00..101888 1144376483 4400..01308643 0 Hsi ou F3408B2r-i2t4t 33 01..047293 1143901982 4400..07473103 00 TsSouuw 5F34i10193W--a22s:4h 333 000...000588758 111533346341128 44400..#.00o35Re49E92N 00 Eo1E ua F5F3S44i00l78l94e---22a44i 33 3 000...000888438 11I33s79i38448s 444000...000533910340 FE Drift 3 1.457 18318 40.7849 0 How Wash 3 0.057 16558 4 #.aEn 0 torw RunOut Wash 3 0.057 16793 4 #. aan o 000276 000194 BE! Calibration curve of SSUSOSAY Fluaride L BESTCOPY AVAILABLE Emm weyrE TES 3.4307187 1 | py l / i A } oy I Order 2 wren r:o0.ssena wl | 000277 000200 [RZ SI a = -------------- Calibration curve of 35050941 Fluoride 1.6 --4 09 ------ ------ : ; I Ss / i 1 Joo C=2 1 J | = i J / /J i|1 -= :i 31 // OIl / I. : i // iI : J #/ i} : re i | | ey 4.0570885 _ ] 0 Heasurad | Crder Inverse Logarithm i| Pl ' "ages | s 5848 000278 000201 " SETSTARTn mewnwesw | 2 -------------------- Rau zt: of 5605093 Fiveride 1.5 --m8------ 1 ngE | BR = R { g{ 14 : 1 I] i 1ih]i J paan do Ri l 2F1ih H ti Pin +LiH Tl - iefd i ii a "i Chl iPiha ffH|it "i bb a f dahHih n! Hil < PPi r ii Li I S~S I1 I- i = bo i tl = fibo | hhHoE ob hi ii iI Hohi no In to HIEoEH ho Ci a HT] oh I e Ha Ho pLHh wdwpffi o loaws 1|] COL ead (BT a] 1pe 3W H A UE E EE CAeMRYAUL YNA SA Y Ai 0. aEN| Tine Ea asExat | FasCrtilPieEdnit pesks | = TT 000279 000202 BESTCOPY AVAILABLE ERTS TE Re TTT IEE Smee --_-------- Raw data oF SENSE Fludride 1.5 ------------------ 4085 - 4od Ed i i i il 4Lo I I | fl it :iiih |! Hy! iJ | | Ed i i| i ii | i i i bi it | iIl Le | PEO 1R Fn I cs"P 7J U PTY LAEfin i 2a7%$4 | LE Pn : So I Tie SeeEKt Fishelp|OrtloeeEdit seme | - oe od Ts 000280 000207 ---------------------- Rau data of 9503324! Fluoride 1.5 ----mm-------- FEES finn,3 riotERRali TheMCC hoton BEI i is,di] 2 2s f "I w 8 ; ; i =2 = . E J \ | => Pi E ! 3 : } i < | ~3 i 3 i] <i0 : || ; | i i i = = | ] i T5e A bi Ra | J TR J ty i 1 { o LTS Tey powa np Ly foe ga Sevier |fte|Gaeicibemndl mes dT 000281 000204 ,- ee Faw fi2a of GE0SC36! Fluoride 4.5 eee ag BE ReRe TEESE eT ITgem en = ag Ss sv o> = i ; y : == : { i E 4 i iI bo 3 i | 31 | i|| qp 1 @ dio FFOaAfALtNpASg tiv hl HNN I Sng wf AL NYY a4 wn me Exit | Fhanals | CebLPeEdi masks 000282 ae 0002075 = 995-0509 15:55 oust ot + ssooms fg oftware : version 5.1 199,33 "yy, serator + Dow Wage ate of the Analysis : 1995-05-09 15:08 nalysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\95050981 "allu%obrriadteio1n.5original time-date stamp : 1995-05-09 09:48 SUIBEAIOR SRIRE Divarvs Sogarions 110pe is = rasan x= x = corrected value of the sample esutt = 10l Ts fC Sloe oi the eledtrode o12I2= 2 -T0.0009 000000 aLluiobrirdaetio1.n original time-date stamp : 1995-05-03 09:48 ststzation order = 2 `orrelation : r = 0.99974 sult = a2 * x2 + al * x + a0 213=21 00ol.lc0oo0os02s00e0 anplor VSyaopmbepelre Time i|+ is50a00s0ec. Masrh TTiimmee [11200se5c. PTReaeekcdeilaeulpHeight : | NSTooi"nneglnem. siuter GaniieTleaudtlteiioonnHeiVFgeahictutmoer |: | 8100 205 mm ml. DReislaumtpiioon runs I|E 1 Ruesperrocfesi.le:| to Cm 000283 000206 . 595-05-09 15:59 output of : luoride 1.5 DSPaeitlghonlaolnrutmyepre :f Dee3busbled SaRiyetsautmeespmlenumber ih0Noo 451 Threshold 4095 dViiGnooutpGuwtent | o0ff s2i sTtaannaasarraa :| Iiggnnoorree sSS33is stSSTTmaaaannnnaaadsssarrrradad :: i IIIgggnnnoooorrrieeeiso 73$5 SSilmraaannnaaasaarrraaa i1 0011.232000000 ODSrlidomeern'sS:iToannIansv+rearEsoei Loga1r3i0t0hm stare value : 500 bu PtiSeriaaiakgrgce'srihioagnpLneoiormreiet :|i1P102o800i0nteSSsdeeaccc HFfoiealgsteeunrzesrswtiinodnow | | hN7eo5 ofourmtuplaut:T we.ees luoride b DpSeiacgtonhtaolnruTmyepre a: Dse0bubbiea aSlytaatceen number |C0o Resample : No Window event | off dil Threshold : 4095 diG output 0 95050981 5,D, 7,2, 000284 000207 A 995-05-09 15:59 OutPut of : sl sTandard : 0.015 s2 sTandard : 0.030 $3 sTandard : 0.060 s4 sTandard : 0.030 s5 sTandard : 0.120 s6 sTandard : 0.150 s7 sTandard : Ignore s8 sTandard : Ignore s9 sTandard : Ignore $10 sTandard : Ignore Order : 2 Dimension : PPM start Value + 500 DU trigger Limit : 1800 Sec Peak shape : Pointed StArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula : c4:=c3 output EER 95050981 000285 000208 bl 995-05-09 15:59 Output of : 95050981 Fluoride 1.5 Pluoride 1 Pen pon os Typ Ident Ch Result P Time tiw InitialTrWaacsehr 33 01..055079 21625 a Drift 3 1.520 386 wUUU FFFSS544400188057W6-a-22s2444h 333 3 0000....000057887803 1067995443154 av DWriafsth 33 10..601597 11520632 Tw Run out wash 3 0.057 1731 Ch Result F Time 4 4 #0..840e6e2n 0 0 4 0.8106 0 444 0#0...0n04n40e34n 000 44 00..08450480 00 44 s#.448s8e8s 00 Page 1 of 1 %, (3) 2 2% %, 000286 nono: ~ 995-05-09 15:44 OutPut of 95050981 Fluoride 1.5 Fluoride PPM PeM os Typ Ident Ch Result F Time Ch Result F Time t Tracer 3 1.490 212 40.7985 wa DWriafsth 33 01..045872 358561 44 0#..7#9#5#1d 00 u F54106-2 3 0.113 751 4 0.0989 0 uua FF55440088D97r--i22f44t 333 001...116500983 11129062126 444 000...108594719763 000 wrw Run out Wwaasshh 33 00..005577 11570337 44 #4..#4#4k4E 00 Page 1 of 1 2%, Ly # ZZz low nebeameL n pdonpao Gun Spokes fraser jets 4a1nd0"raMpartmveSiatlyasrQeonm|sreecd 000287 000210 / SEEses Teme prey ------------------------------ Raw data of S505098t Fluoride 4.5 as + Sn - 1| iy | i i Til : -------------- Sees SEs 1 = CE= = = i - = Pi = hii I 1 i i H Iii 3 Hit it II E IEl | JmHyi i!i I| i HHiH i i :Tih 1 i | Id 5 | Fa Te I it lA y rAFNh A ltoawnvaodlyusmies L--aaite4SESaunkgalsaosxfe | o d | VE = eST oRndelrStrenAtmS omSaA Herat) 0 Tine Sao SC |FieE Rals | Crti-peait peaks | TT 000288 000211 Le 995-05-11 13:19 OutPut of ': 950511A1 LAorwpT"lHarzolaass. esas oftware : version oon 6.1 1990,93 29-159 Sew BEST COPYAVAILABLEg luoride 1.5 alibration order = 2 Wash Time : 120 sec. User file : . TXT 000289 000212 9395-05-11 13:19 OutPut of : lnoride 1.5 PSaitghnalnumtbypeer DeSycsotleonr Number aiLute dRielsamTphlreeshold 4iG output Window event i: De3bubbled :: Y0es to i: N4o095 i0 : Off 5512 ssTTaannddaarrdd :: IIggnnoorree sS53i5 sssTTTaaannndddaaarrrddd ::: IIIgggnnnooorrreee 5567 ssTTaannddaarrdd :: 00..310500 58 sTandard 0.500 s5910 ssTtaannddaradr:d: 11..250000 ODsrtidamerertnsiv: oanlIunsv: erPsPeM Logarithm 500 DU trigger Limit : 1800 Sec PSeNtedAarktshiiaggpnneoorree ::: P6102o0inteSSdeecc Measure window : 75 1 ` FRfieolgrtemeunrlearation :: Nfioo output: #4.### luoride L PSaitghnalnumtbyepre : De0bubbled SDeycsotleomrNumber `: N0o dilute : No aRielsamTphlreeshold :: N4o095 diG output i0 Window event : Off 950511A1 ; WeyPy, Ay, Ugg 000290 . 000213 ~ 995-05-11 13:19 Output of : ssi2 ssTTaannddaarrdd :: 00..001350 $3 sTandard : 0.060 s4 sTandard : 0.090 s$56 ssTTaannddaarrdd : 00..112500 57 sTandard : Ignore $8 sTandard : Ignore $$910 ssTTaannddaarrdd :: IIggnnoorree order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed eSNtdart iiggnnoorree i: 16020 SSeecc Measure window : 75 % RFeigletenreration :: NNoo formula : c4:=c3 output i #.###H 950511A1 000291 000211 995-05-11 13:19 Output of : 950511A1 sexFluoride 1.5 oonFluoride L `os Typ Ident Ch Result F Time t tiw InitialTrWacaesrh 33 10..407527 20685 wd DWraisfht 33 01..045772 832842 s521 SSttaannddaarrdd 21 33 00..006740 791239 53 Standard 3 3 0.085 1085 s4 Standar4 d 3 0.107 1260 s5 Standard 5 3 0.127 1435 ` 0 ss76 . $8 SSttaannddaradr7d6 33 00..218593 Standar8d 3 0.607 11671804 1959 32 5910 StSatnadnarddar1d00 33 11..426383 22310393 54 oawd DWraisfht 33 01..055079 22742854 75 oouu FF5544018123--4488 33 00..009914 32083161 su F54111-48 3 0.095 3185 9 wu F54098-48 3 0.091 3359 I0 uwu 2 ou FF5544007894--4488 FS4116-48 333 000...101080123 333875813717 3 u FS54113-192 3 0.097 4061 554 auuw FFS554401812D1-r-1i19f92t2 33 3 00..008819 1.435 44421337 4585 87 uow F54098W-a1s92h 33 00..005878 44973362 09 uu FF5544007894--119922 33 00..008894 55121815 zLuu F5544111136--3159620 33 00..008855 55643588 3u F54082-360 3 0.073 5810 4u F54111-360 3 0.087 5984 76Jouuu FFF555444000987849---333665000 333 000...000998122 666513163346 35 wa DWraisfht 33 01..045871 66962865 i0 uu F54116B-LK3601 33 00..016059 77201339 32 ouu 3u SSPPXK 21 F54082-D22 33 3 o0i.109142 0.085 77336857 7739 SSuu FFS5544017191--DD2222 33 00..008834 87098152 37 uou F75544101352--DD2228 33 00..008873 88423683 05 au F54111D-rDi2f8t 33 01..057189 98768147 1 ow Wash 3 0.057 9027 25 uuw FF5544101769--002238 33 00..008717 99311348 Ch Result F Time 44 0w..7a3w4s2n 00 44 0#..7#3#4#2# 00 44 00..00218674 00 4 0.0586 0 4 0.0921 0 4 0.1194 0 44 00..12469482 o0 44 00..46258257 00 4 0.7329 0 44 0#..7##4#746 00 44 00..00678252 00 4 0.0758 0 4 0.0685 0 444 000...000588264334 000 4 0.0775 0 44 00..00561329 00 44 #0..#7#2#1#3 00 4 0.0626 0 44 00..00673299 00 44 00..00558753 00 4 0.0353 0 4 0.0611 0 444 000...000566198991 000 44 #0..47F3#7E4 00 44 00..00915898 00 44 00..10070272 o0 44 00..00556896 00 440.0.00553460 00 34 00..00464077 0o 44 40..7454140 00 44 00..00540321 00 Page 1 of 2 2 2, 22 Z 0002175453 000292 995-05-11 13:19 OutPut of Fluoride 1.5 Pry. os Typ Ident Ch Result F Time 45 ouu FF5544009999-84 33 00..007710 99469602 3a F54099-12 3 0.105 9840 357 oouuu FFF555444000999999---1243482 333 000...100187844 111000013136502 O1? uu F5544605999--336604--336-00.05770 T1O0T5E38S z5 aow DWriafsth 33 10..055074 1110182808 iu Ps4112-4 3 0.082 11237 sTsuuu FFSF545414l111212-2-2-1482 333 000...010908408 111111457196515 33 ouu FFS544111122--14982 33 00..100904 1112914115 Oiuu F54SP1K12-6336-01 33 00..007985 1122248695 z5 uou SSPPKK 6633--23 33 00..101902 1122861871 15 aow DWriafsth 33 10..045673 1132196859 5ou SP 126-1 3 0.117 13342 7 uw SPK 126-2 3 0.149 13518 o33 wua SPK D1Wr2ia6f-st3h 333 010...104559776 111433168096725 rw Runout Wash 3 0.057 14340 : 950511A1 Page 2 of 2 Fluoride L PEM BESTcopy, Ch Result F Time Vilage 44 00..00239187 00 4 0.0834 0 444 000...010305776153 000 4 0.p03r06t------00-- Bglant plingaw sbed h=na e Sn ue 44 0#.47448547 00 4 0.0527 0 444 000...000786322574 000 44 00..00871255 00 44 00..00475520 00 44 00..00967833 00 44 0#..47#3E3H4 00 4 0.1061 0 4 0.1452 0 444 00#...17#544328774 000 4 #.#### o 000293 000214 EER enemy ome | BY ---------------- Calibration curve of 3E0S1HA Fluoride L ee 1729504 1 "4 : = :# =BE= | cose _ order 2 ; .33887 000294 000217 Calibration curve of SOSH Fluoride 4.5 ------------ rasrss WERTHTeer ee i a=3 ||| / i = = = } / ii -=< i / | il / | ! / ' sessed | i o Grdar Inverse 'LosarithnHessurad ' FRE 033 000295 000215 eu Sits oF BEES Flori tf BETESI ues TERpms saeSs | Va >a w FH fg =i i: IE ih | [I i ii i [i1] B 3 To hi i 1 Ti 3i : il Hi Hitil i wHihIliEih HE dh h hin ol Po HH iifi||o |i| i | HE J| e aLi dil i i 1 HilSaga v Ygmn <i} s Apia RECA RS E yy AAA RRRE $e PPSaA JAAU YURY INE ULMYNVYUV WUU UYEEl LI ) Tine zone Securit Fishain|OrtibpeeEwsa1it TTT 000296 000219 1 ! lo ------------------ R2u data of 35051181 Fluoride 1.5 -- EEReE yBakes Tir IRIE armsass vs |* 2% i | : 4 i |! !| iI |I If i } 1 i | I| if| I | 4 iil i iH H1i i| 1 il I i i ||| Il| I| i| || 1 1! 1Te Tan am ww wa0 || || DE i FI) Tet si) F aAR ATU R f{ al ITA P usSail lei Rws U nAG ARRA wS R | -_-- 45E0 Tina sasExit | Fiohelp | Crii-Peedit peaks | BE } 35680 I 0002997 00022i] 0 = flay datz of 5054434 Fluoride 4.5 --0no aaeys 2 etSS SHRTTDeerSea a "A "| i1 | ii i i Jl II |i ii i il i itl 1i1 ii} | i iii i [| HE i io] i ii Hnhi :i ii iti fli 3 I I Ii | 1 I i ! Li 1} ;| | i adih J| 5} HH RE ale 3 #1 Ds wate Sell HAH Pow ahdde 001 11H C FPAA a UNRs AR v TaLg Be eH MNl M Y LU I e N VphYA d Y In Yii I s 3305 tee "tip netS e grerees eeetereteet r 3935 | se=Exit | FisHelp ! Crtl-P=Edit peaks ! 3'EST COPY AVAILABLE 000298 | 000221 Py a =-------------------------- Rzu dats of 9505i1A! : Fluoride 1.5 ------------ BEETR r S Re TEA Em ws eee i fi ii | ot | it it il il | | ! il | i it La a FIRE Ae i 1 le ii Hid RATAUERNAURESAN] --_-- o = 12426 Tine i I: | |Li I i | I| } 1 I 174351 BEST COPY AVAILABLE 000299 000224 Oow "lazlas 995 05 24 16:00 OutPut of : 950524A1 ATnadTaT-s4a2095. Jitwars : version 6.1 <19$90,93 perenne cow BEST COPY AVAILABLE ate of the Analysis : 1995-05-26 11:39 nalvsis File Name C:\SKALAR\DATA\WIDATA\SERUM\35052:A1 BLuortidei1.5enter = turers Togsrinin ope cs = marron ameh aty] RB cl = = cooorrreeecutieedd vave valus of of the the sample concentration 1 asult = 10 s s = Slope of the electrode 5i21= -D0T20.ai000s00e006s04 sLluiobrirdaetion order = 2 orrelation : r = 0.99786 sult = a2 * x + al * x + 20 51212- -0T00o.0l0o0000800280s smpier MTyepmeber irsa1000 VmSTaaaeskmehpleupTTTiiemmeee :}iV5S102i0E msseeecc. RSerseaclieatHaight || NJoonamm Lurer ldnieuletuditleioonnHaiFvgaohcrttuomrs|:| 3210605 am ma. Driolpeteenn runs 0i1i Ueserr ofile:i 1 Ce 000300 00000:227) 995-05-24 16:01 output of : luoride 1.5 Path number : 3 SBSHeivElgsonctaeelnr Ntuympbeer + Debubbied vfoes aRe1sanTshiresenota | 4408s diG output i0 lurid L Window event : Off 521 sSttaannddaarrdd |: ITgonnoerres sS553i55 SSsSTrTTaaaannnnddddaaaarrrrdada :| IffogonnnoooTrrrseee o 5573 Traanndmaa i 00.38030 sS9lo SsTtaannddaradrai: 11.520000 ODStriadmreetrnsiV:oanlIunev: |erpso:ei Lo50g0ariotuhm tPSSheriaaiAkgrgcesirhisadhpnLoeoirrmeei|t :| 1Pg1o28o.00i0nteSssdeeeccc HFFeelasismeumrreerawtiinodnow | | i7e5 Ne 8 cfaorpnuutia | wnans pSaltohnalnumTbreers : D0obubbied DSRieevcsseeatelmoeopnrleHumber +i| hN0looe adiilc Touhtrpeusthold 1| 0409s Window event : Off 950524A1 BES, TCopy py, 000301 000224 9595-05-24 16:01 OutPut of : 51 sTandard s2 sTandard : 00..003105 $s43 ssTTaannddaarrdd :: 00..006900 $5 sTandard : $6 sTandard : 0.120 0.150 $7 58 sTandard sTandard :: IIggnnoorree $9 $10 ssTTaannddaarrdd :: IIggnnoorree oDridmeernsi: on2 : PEM sttrairgtgervalLuiemit : 500 DU : 1300 Sec Peak shape StArt ignore : Pointed : 80 Sec eNd ignore Measure window : : 120 75 Sec 3 Filter Regeneration : NNoo formUla : c4:=c3 output: #.##4# 950524A1 000302 0002275 595-05-24 16:01 outPut of : 950524A1 Fluoride 1.5 Fluoride oo eon os Typ Ident Ch Result F Time :iw InitialTraWcaesrh 33 01..403529 21555 waSi standaDrWrdiafsth1 333 001...004856399 763428109 5S3z Si Ssttaannddaarrda 3z Standard 33 3 001.008773 0.109 1093106 1269 33 Stanaara 5 3 0.127 1442 5 ss37e 3S sSStttaaannnddaadrraadr7de6 333 o000...21e85145z Standar9a 3 1.242 111976831508 2140 P5FE wS10 StandarDdrif1t0 Wash 333 011...044554982 22433104 2128 iiaa su BBLLKK12 33 00..007858 SPE 63-1 3 00103 3Z0a1451 3191 3a. 1a SSPPKK 6533..32 33 00..003388 SPK 63-4 3 0.093 33333635 3115 EFR N ia SSPPKK 6633--55 33 00..110024 SPK 63-7 3 0.101 43088371 4243 Ts5 qwu DBWLrKiafsth3 333 010...040537952 84434331115 353 ooouuu I 53430089B89L-.K445458333 000...000778138 55089.43 3 0.085 43539214331e90ss zu 31007-45 3 0.085 3644 6i3 uuoau FFF3354s4100098920.--444588 Fsilli-ds 333 3 000...000786057 0.070 s3598i81e82 e344 s37 waou 5u F54088DW-raDiYsf8th 333 010...040586358 8e6955331027 54099-D78 3 0.085 704i FF1RI u I SPSSXPPK6386-331--083 333 000...011900901 BLsi 3 0.073 7777733249811338 ssau Ta Se BL8K3-12 SP 63-2 33 3 00..005858 0.058 878290613753 su SPK 53-3 3 0.102 8413 HD3[Ia Uou F54039D-rWDiaYfs8th esi0s7-Ds 333 3 001...004577520 0.075 2898176043123393 u F54080-D3 3 0.075 9319 Ch Result F Time 440.n4.e0a8n7e 0 4 44 0#0...0#41n08k50E4 0 0o 44 00..00257689 4 0.0945 0 0 4 01182 0 4440.00..251444139718 I 44 00..44003576 00 440.#.4404548% 0 4400..00138380 00 440.0.00788413 0 444000...000687888308 00 440.0.00882219 00 44400.#..40402956084% 00 444000...000212337383 40.0110 00 40.0102 0 44400.0..000201134851 o0 40.0328 0 44400.#..40a01r55a28r 00 40.0170 0 444 000...000778791402 000 4440.00..000082897378 00 0 43 00..00373683 44440.00#..0.043#2093458n74 o0 4 0.0323 o Page 1 of 2 %. 2 2= ", =z 000226 000303 395-05-24 16:01 OurPut of : 950524A1 Page 2 of 2 oo son AVAlLapLE Fluoride 1.5 Fluoride BESTCOPY 5 Tp tdent Ch Result 7 Time DLuu Brsiesazosve 33 o0.n07s7 s84a9r3 DD3 U0ou ERF53S45i10o1%s4aa-0bDiiis3 333 000.l0007137 G1o09t08s54:33 [J J0u U PBF53i40o088i09o-0Dii1ss5 333 oo0.ll06ss8 il1o0os3ni7ss0 D2Ua- esernDaaraisifhtt 333 001l..004%56039 i11i10s184393i3 Pa F54109-D15 3 0.072 11418 JD75 u4Wou EPEF5Ni4a0o18s1l47a-o0bD3a1s2s5 3333 oo0o.lll0oe6srs8 ai1i1iti5se9ssr4 Ju F54114-D22 3 0.069 12296 Lu F54089-D28 3 0.066 12466 HPD2 du-uw HF5o40b8D7ari-iDsfe2ht8 3333 05o0..d00s6e%8 111s233a63340s32s DU rsawetdis 3 ols iasis ITu PKSPK 86331-54 33 o0.l0a7y8 13s 13515 1Pjiudu EssLeSSlPIoKksT-e6s3Esa-m6 3333 0oo0.lo0de9ns8 1iI h3i8sa7s2 DH3i dL nunoutGWWraaafssthh 333 00a..i00i9% illisasostsso Oh Result F Time 440o.l0o3m6s5s 3o 444 00#.00973669s5 o0o 444 000.000110i675 ooo 444 003l..044n08i58e48 o50 4 0.0249 0 4444 o00ol.lo0loi1ios4sre6e a3a0 4 0.0170 0 4 0.0114 0 1444 no00.4.i0c0n1e16es08 aao0 4 0.0160 a 44 00l.00%31709 a0 ii14 o0o0l.n0o0i7:%s6o3 :ao0 14 pio.idb3e9ess :oa 000304 000227 I BESTCOPY AVAILABLE 000305 000225 mere S2EliprEationECurr e 37 HSEe R | FIUBAIa OE 1.5 mmr mroennn ; 4 Fl i / tbih // Li o 1.655575 enti bo ] epee epee me eee penne ee. | | siessurad ' ams BESTCOPY Ayal gp, 000306 000229 ee | ASAT bg L8 osoBnEnT ATFRmnOeMnErFeE1.t 8p mee o et m rt enaeo Ia I. PE| i i tah > vo Hi w HHit i i w | Co | bo i|i Bi i i HH wo HH bi il |} .a ii | i IRR 21% ] CL i i} wha eae Rea Io || i 14| 2 SEY F|l Gelieeda attmes TTT BESTCOPY AVAILABLE 000307 000230 ---- IEE War Eers -------------------- Rau data of $50524=1 Fluorios 4.5 ---- -- Ey - ke i ! i :1 i |i| LoI! i 3 i i !" | ra Loh oe Tam patina i . T2E sasEat |Flake a |OrbipgeeEmsmic TT BESTCOPY AVAILABLE 000308 0002711 me a 7313 3 BHR Frieris 18 me oomm w EX i SINS Qa Sap A A pT Tw NER " at BESTCOPY AvaILgpy 000309 00207 0 kd 9 se 5 ee Ae % oF? ddd * SANE Ag x ve oaothnlas BEST COPY AVAILABLE 000310 0002 BHAZLETEN MARLEEN Sponsor: 3m St. Paul, Minnesota FINAL REPORT +CORNING Conon FECEIR Cee SEP 121935 2M Qeicor0? Study Title: Single-SDtousdey IonftrTa-v6e2n4o6usinPhRaarbmbaictoskinetic Author: Steven M. Glaza Study Completion Date: September 14, 1995 Performing Laboratory: Ha33z0l1etoKninsWmiascnonBsoiunl,evaIrndc. Madison, Wisconsin 53704 Laboratory Project Identification: HHI 6329-159 EPXhPoRnEeSS-6M0A3I:LisiDiELsIaVlEiR_Y Page 1 of 38 3301 KINSMAN BLVD 000311 MADFIgSaOoNxs. :2waii|:152372074 Page 2 of 38 HHT 6329-159 QUALITY ASSURANCE STATEMENT TW(hiFisDAsc)onrseGiponoo,drt LIahnacb.so,rabteioenrnyarcecPvoriraedcwatenidccee bwyRietgthuhleatthQieuoanlFsio,tody21AasnsdCuFrRDarnu5cg8e.A3d5Umnii(ntbi) sotf(r6a)Htaiz(ol7ne).ton TSanhtdeudyfFoilDnlidoirwneigcnstgorarieannsdpiesmcsatuneiadognestmoewneHtra.ezlectWoonrndiutctmteaenndagsetamnadetnutsfinmrdoeinpntogrhstlsyreoapfcocroitrnedsdipnegctotitootnhse standard operating procedures. FInrsopnection DTaotes Phase RepoDratteed to Date to Study Director Management 0038//2120//9955 0034//2120//9955 PPrroottooccooll ARmeevnidemwent 0047//2075//9955 0047//1201//9955 ADnaitmaa/lReOpborstervRaetviioenw 09/12/95 09/12/35 Report Rereview 0043//1202//9955 0074//1207//9955 09/12/95 0053//1100//9955 0058//1100//9955 10/10/95 Represeftative, Quality Assurance Unit F.14-95 Date 000312 Page 3 of 38 STUDY IDENTIFICATION Single-SDtousdey IonftrTa-v6e2n4o6usinPhRaarbmbaictoskinetic HHT 6329-159 Test Material Sponsor Sponsor's Representative Study Director Study Location StuSdtyudyTimIentiatbilaetion Date IExnp-eTriifmeenEtnadl Da(tIen-Tife) Start Date SEtxupdeyrimCeonmtpalletiToenrmiDnaatetion Date T-6246 TMonxicology Service 3MMeCdeintcearl, DeBlpdagr.tme2n2t0-2-02 SPt..0. PaBuolx, 3M3N22055133-3220 J3oMhn L. Butenhoff, Php ToMxeidcioclaolgyDeSpearrvtimceent 3PM.0.CenBtoexr,332B2ld0g. 220-26-02 S(t6.12)Pau7l3,3-1M9N62 55133-3220 HStaezvleentonM.WiGslcaoznasin, Inc. MPa.0d.isoBno,x 7545 WI 53707-7545 (608) 241-7292) H33a0z1letKoinnsmWainscoBnosuilne,varIdnc. Madison, WI 53704 AMparriclh 34,0, 19199595 Staeypte1m,be1r99514, 1995 September 14, 1995 000313 Page 4 of 38 Acute Toxicology Steven M. Glaza Study Director Manager Francis (Bud) W. McDonald Study Coordinator Patricia Padgham In-life Supervisor RReopsoertM.. SuBrpiedrgveisor Toxicology Support Kathy Myers Manager Calvin L. Horton Supervisor KEY PERSONNEL HWI 6329-159 uality Assurance Sherry R. W. Petsel Manager Laboratory Anima Medicine Cindy J. Cary, DVM Diplomate, ACLAM Supervisor AnatomicalPatholoay Thomas E. Palmer, PhD Anatomical Pathologist Jack Serfort/ Deborah L. Pirkel Supervisors Necropsy ASnunpeerMvoisshoerr Pathology Data 000314 Page 5 of 38 CONTENTS QSutauldiytyIdAesnstuirfainccaetioSntatement SuKemymaPreyrsonnel ObRjegeucltaitvoery Compliance TTeesstt SaynsdteCmontrol Materials PRreoscueldtusres DSiisgcnuastsuiroen Pathology Report Table 21 IInnddiivviidduuaall CBloidnyicWaelighStisgns(g) 34 IInnddiivviidduuaal) APnaitmhaollogTyissCuoemmeWnetisghts and Bile Volumes AppPernotdoicxolA Deviations PPrroottooccoo]lTPABm0e8nd4m.ePnKt No. 1 HUI 6329-159 Page 32 46 88 8 109 1133 114s 1186 221 225 3277 000315 Page 6 of 38 SUMMARY HHI 6329-159 aThdimsinissttuedryedwasbydoanesintgoleassienstsravtehneoulseveilnjoefctisoynstetmoicrabebxiptoss.ure of T-6246 when tThhee Hsrtau:d(yNZwWa)sSPcFondsutcrtaeidn ufosringeacfhourtrmeaaltemenatndgrfoouupr afsemafloelloawcsc:limated rabbits of Group 1 (Control) 23 ((LMoewd)ium) 45 ((HHiigghh)) Test Material StTe-ri6l2e46water TT--66224466 T-6246 Do_s(ema/kLae)vel 0 105 510000 MNaulmebser of AFneimmalaelss 44 44 a4 44 a 4 * Opnoessiabnliemalbroskaecnrifbiaccekdanadftreerpltarceeadtmweintthduaenotthoera male aninal Tthhee ainnidmiaclastedredcoesievedTevaelsinignlteo tihnetrmaavregnionuasl ienajrectvieoinn dose volume was 0.5 ml/kg of body weight for Groups ooff tthhee rtiegshttmaeatre.rialTheat 1 through 4 and was Da0an.yd75t28hm.el/krgemafionrinGgrouapnim5a.ls"T(wtowoanainnailmsa/lsse/xs/edxo/sdeoselevleelvelw)erewerseacrsiafcirciefdiceodn Donay 15 Ct4lwiihncoieucrasal daaofybtseemrrovriatntatilroiantvsyenowcuhesercekiscnojnwedecrtuiecotnec.dondpuArdcedtdieotdsieodnaaainldlyactltihnaeiprcpearalofxtieomrbasteuenrtlviyalti0o.t5nh,se 2a,ndand oasndcmhieDndayiulset-dr7atsfioaorcnrirfa(inDcadeyomii1z)n,atteiraovnandl patu(rDpatoyhsee1s5s,cohrebdeufDloaeyrde28ts)ea.sctrifoBirocdeyconWitenritoeglrhvtasmlatwee(rrDieaaylde1t5eromrined Deaayr v2e8)i.n (Alefbtlooedar)saompflethe(aapnpirmoaxlismatately4-,4 m8L-), post-injection, and on Day 8. An approximate w1a2s-, co24l-l,ectaendd 4f8r-omhouarsmarginal 4-nl blood sample was also cIonllaedcdtietidono,n Daatysthe15tiamned 2o2f tfhoer tshcehedaunliemdalssacsrcihfeidcueled(Dfayor15sacorriDfaiyce28o)n, Day 28. caepnptrroixfiumgaetd,e"ly se20panrlateodf Sponsor. ' On Day 15 or bilnotoodswearsumobantdainceedlluflraorm efarcahctiaonnism,al.andAlslentsamtopletshewere 28, the animals were anesthetized with sodium pacoeblnbltreoecbvtaieradbt.ietdalTg,rhoesbswlheodnleecvriaolpisvteyhre,expaboimslitene,artiioaornnd vwebanosath cnoaktviad,dnoenyaesn,d fheroxowsmeavneegaruc,hinaattnieisdmsa.uleswAenwreere cSpoolnlseocrt.ed, weighed (volume only determined for bile), and sent frozen to the 000316 Page 7 of 38 HIT 6329-159 ctIhhnartnorguaegvsheonuoituns tbhoiednyjsewtceutidigyohntwiotfgahiTn-.t6he246AelxlcdiedpatniinonontalsorfesaupolnpteearGierndouapcnlyi4 ntieascntdallmonyaetenroGrirmaoaullp-re5lamtaelde aa(nnpiiommsaasllissbletahpapbteraorkweeednrecblasicakncsir)ci.aflilcyeTdhensoorenmaDalanyismtahlr2souawgnehdroeu1,t rterhpeelsapcescettduidyva.enldy,theduereptloacienmjeunrty 000317 Page 8 of 38 HHT 6329-159 OBJECTIVE Tthhee otbejstectmiavteerioafl,thiTs-62s4t6u,dywhweans taodmiansisesstseretdheaslevaelsinofglesysitnetmriacveneoxupsosuirnejecttoion to rabbits. REGULATORY COMPLIANCE ATdhimsinistsutdryatwiaosnscoGnodoudcteLdaboirnataocrcyordParnaccetiwciethRegthuelatU.iSo.ns Fofoodr and Drug Nonclinical mLiaxbtourraetsoryforStucdoinecse,ntr2a1tiCoFnR, 58h,omowgitehneitthye/seoxlcuebpitliiotny,thaatndansatlaybsiilsityofwatshenottest AcnoinmdaulcteWde.lfaArlel AcptrocReedguurleastiounsse.d inInthtihse osptiundiyonweroef tihnecoSmppolnisaonrceandwitshtudtyhe director, the study did not unnecessarily duplicate any previous work. TEST AND CONTROL MATERIALS Identification Tlhiequitde.st mTahteercioanltrowlasmaitdeernitailfiewdasasSteTr-i6l2e46WaatnedrdefsocrriIbnejdectaisona, clUeSaPr,(Acboboltotrless Laabcloeraart,oriceosl,orlLeostsNol.iqu8i6d-.748-DM-02; Exp. April 1, 1996), and was described as Purity and Stability TdheeterSmpionnastoironasssu(miensclurdeisnpgonsuinbdierlittyestforcontdeisttiomnast).erialAnapluyrsiitsy and stability of the test mnoattercioanlducmtiexdturoersrefqouresctoendcenbtyratthieonS,ponhsoomro.geneTihtey/psuorliutbyilaintdy,staanbdilisttyaboiflittyhewas csotnutdryo.l material were considered to be adequate for the purposes of this Storage and Retention The test material was stored refrigerated. Asntyoreudnusaetd roteosmt tmeamtpeerriaatlurew.as rTheeturcnoendtrotlo tmhaeterSipaolnswoars SatfatnedrarcdomOplpeetriaotningof Pralolcediunr-eTif(eSOPt)e.stinAgny acrceomradiinnigng tovehHiaczlleetomnayWibsecounsseidn fo(rHWI) other testing and will not be discarded after issuance of the final report. 000318 Page 9 of 38 Safety Precautions HWI 6229-159 Tahned ptoelsitciaensd. control material handling procedures were according to HWI SOPs Test Animal TEST SYSTEM Adult albino rabbits of the Hra:(NZW)SPF strain were received from HRP, Inc., Kalamazoo, Michigan on March 8, 1995 and maintained at the Hazleton Wisconsin facility at 3301 Kinsman Boulevard, Madison, Wisconsin. Housing After receipt, the animals were acclimated for a period of at least 7 days. During acclimation and throughout the study, the animals were individually housed in screen-bottom stainless steel cages in temperature- and humiditycontrolled quarters. Environmental controls for the animal room were set to maintain a temperature of 19 to 23C, a relative humidity of 50% +20%, and a 12-hour 1ight/12-hour dark lighting cycle. In cases where variations.from athdevseerseconedfifteicotnson extihsetesdt,udytheoyutcwoemree. documented and considered to have had no Animal Diet TLhaeboarnaitmoarlysRawbebreit prDioevtideHdF a#5c3c2e6s,s tPoMIwaFteeedrs,adInlci.bitTuhme anfdeeda miesasruoruetdinealmyount of analyzed by the manufacturer for nutritional components andenvironmental contaminants. Samples of the water are periodically analyzed by HWI. There were no known contaminants in the feed or water at levels that would have interfered with or affected the results of the study. SeleofcTetstiAniomanls The animals were identified by animal number and corresponding ear tag and were placed into study groups using a stratified body weight randomization program. The randomization body weights were determined on Day -7. The weight variation of the animals for each group of each sex selected for the study did not exceed +2 standard deviations of the mean weight, and the mean body weights for each group of each sex were not statistically different at the 5% probability level. 000319 Page 10 of 38 Study Desian HHI 6329-159 Ainnitomaltshewefioglhlionwgingfrosmtud2y,52g3routpos:3,276 g at initiation of treatment were placed Group 21 ((LCoown)trol) 43 ((HMiegdhi)um) 5 (High) MTaetste/rCioanltrol StTe-ri6l2e46water TT--66224466 T-6246 Dosnea/kLgevel 0 105 150000 VoDlosueme mi/ka) 0.5 0055 00.575 NMaulmebser of AFneimmaalless* 4 4 44 44 aa 44 * Obnreokeannimbaalcksaancdrifriecpeldaceadftweirthtreaantomtehnetr mduaeletoaniamaplo.ssible a Trweomaianniinmgalasn/isneaxl/sdos(etwolevaenlimwaelrse/sesxa/cdroisfeiceldeveoln) Dawyere15. The sacrificed on Day 28. Justification for Species Selection cHhiositcoericbaelclayu,setohef Ntheew ZlaeragleandamoWuhnitteofalbbiancokgrroaubnbdit ihnafsormbaeetniontheonantihmiasl spoefcies. PROCEDURES Dose Preparation and Administration cThoencetnetstratmiaotenrifaolr weaaschdidlosuetedlevweilth insteGrrioluepswa2tetor to 5. acAnhieivnedivaidsupaelcidfoisce of oenachitsresbpoedcytiwveeighttestonsothleutidoany oorf ctornetartomlentw.as Tchaelcurleastpeedctifvoer teeascth asnoilmuatlionbawsaesd eaadrmionviesrteraepdprobxyimiantterlayven2o2ustoi6n0jecsteicoonndsi.ntoThteheprmearpgairneadl teesatr vseoilnutioofnsthweerreight rsetmoraeidninagt rteosotm tsoelmupteiroantsureweruentidliscaadrmdiendi.stered. After administration, any Reason for Route of Administration Intravenous injection is an acceptable route to assess systemic exposure. 000320 Page 11 of 38 Observations of Animals HUI 6329-159 4Clihnoiucrasl afotbesrervianttiroanvsenowuesreicnjoencdtuicotn.ed pArdeddiotsieonaanld caltinaipcparloxiombasteerlvyati0o.5n,s twice a day mortality checks were conducted daily thereafter until the a2,nd and scheduled sacrifice interval (Day 15 or Day 28). Bteosdty woerigchonttsrowlerematdeertiearlminaeddminonistDrayati-o7n fo(rDayran1)d.omiAzdadtiitoinonpaulrpobsoedsy determined at the scheduled sacrifice interval (Day 15 or Day 28) waenidghbtesforweere or at unscheduled sacrifices (when survival exceeded 1 day). sample Collections A(lebfltoodears)ampolfeth(eapapnriomxailmsateatly4-4, mi8)-, wa1s2-,col24l-e,cteadndfr48o-mhotuhresmaprogsitn-ailnjeecatriovne,in aanndd o2n2 Dfaoyr 8t.he Aannimaaplpsroxsicmhaetdeule4d'mLforblosoadcrisfaimcpeleonwasDayals28o. colInlecatdedditioonn,Dayast t1h5e tviemnea ocfavanecorfopesayc,h aanpipmraolx.imaAtlellysa2m0plmlesowferbeloosdtowraesd oatbtarionoemdtefmrpoemratthuerep,ostanedrior wtehreen ctheenntrisftuogreedd, insepaarfarteeedzerintsoetsetorumnaianntdaicnellaultaermpefrraatctuiroenso.f -T2h0eCse1s0amCples until shipped to the Sponsor. Pathology Tnheecroapnsiimeadls insacthreifiscaemde moannnDearysas1 tahned a2nidmuaelstosuarpvpiavrienngt tboroktehne bsacchkesduwleerde esxaacnriinfiacteisonwiatnhd tahney eaxbcneoprtmiaolnititehsatwetrheeyrerceocredievde.d an abbreviated gross necropsy OgnrouDpayas15s,igntmheentfirrsatndotmwiozaatniiomna)ls/wseerxe aasnseisgtnheedtizteodeawcihthdossoediuTmeveplent(obbaasrebditoanl the a(nvdiaexisnajnegcutiinoanteidn. thAenmaarbgbirneavliateeadr gvreoisn)s, nebclerdopsvyiaextaheminpaotsitoenriowarsvennoat dcoanvea,, heaocwhevearn,imatliswsueerse wcoelrleecctoeldl,ectweedi.ghedThe(vwohloulmee olnilvyer,detbeilrem,ineadndfobrothbilKei)d,neyasnd from Aifmtmeerditaitsesluye/pblialceed coilnlaectfiroene,zerthesetanitomalmsainwetraeindiasctaermdpeedr.aturTehe ofrem-a2i0nCing10twCo. athneimaslasme/semxa/ndnoesreaslevtehle awneirmealasnesstahcertiifziecde,d bonledD,ayan1d5. exsanguinated on Day 28 in 000321 Page 12 of 38 Shipment of Tissues HUT 6329-159 Affrtaecrtiocnosm)p,letliiovnerso,f tbihlee,ina-nldifekidpnheasyes,wetrhee bselnotodfrsoazmepnles(on(sderryumiacen)d ctoellthuelar PSapuoln,sorMN,(J5a5m1e0s6)D,. Jaolhonnsgonw,ith3MthE.eEi.r c&orP.rCe.s,ponBdlidgn.g w2e-i3g-h0t9s, or935volBuumsehs.AvenTuhee, St. HSHpIondsooers isnotresapcocenpstiblaenyforrespthoensribeitleinttyionforandthedisapnoasliytsiiosnofofthtehesasmapmlpelses collected in this study nor are these results presented in this report. Statistical Analyses No statistical analyses were required by the protocol. ocation of Raw Data, Records. and Final Report Trheetairnaewddaitna,therecaorrcdhsi,vesandofanHUIoriigninaaclcorsdiagnnceed cwiotphy oHAfI thSOeP.final report will be 000322 Page 13 of 38 RESULTS HHT 6329-159 Body Weights bIonddyiviwdeuiaglhtbogdaiynwediugrhitnsg atrhee isntudTya.ble 1. There was no meaningful effect on Clinical Observations tIhnrdoivuigdhuoault cthleinisctauldyswiigtnhs tahree finolTlaobwlieng2.exceAplltioannsi:mals appeared normal OhnaeveGrionujpure4dmailtes b(aNoc.k oFn541D0a6y) 2tranedatweadswistahcriTf-i6c2e4d6, atnec1r0o0psmige/dk,g aanpdpeared to rtehprloaucgehdoutwitthheNos.tudFy5.4099. The replacement animal appeared normal + OhanveeGrionujpure5dmailtes b(aNoc.k Fon540t7h6e)datyreoaftedtrweiatthmenTt-62a4n6d waats 50s0acmrgi/fikcgeda,ppeared to nnoercmraolpsitehdr,ougahnodutretphleacesdtudwyi.th No. F54112. The replacement animal appeared Pathology aInnidniavlidutailssuaeninwaelighptasthoalndogybilceommveonltusmesarearepreisnenTtaebdle i4n. TabTlhee n3.ecroIpnsdyiviodfuaalnimal dNoi.ffuFs5e4l1y06darrekvearelde.d thIen asnkiemlaeltalNo.musFc5l4e076s,urrtoheundsikneglettahle mluusmcblaer sspuirnraolundcionrgd was rbeotmhainhiinngd Tainnibmsalsandsurtvheivesdacrtaol thspeiinralscchoerddulewdassdaicfrfiufisceelyadndarkwerreed.notTheexamined grossly, although tissues were saved. Page 15 contains a pathology report by the study pathologist. DISCUSSION TahlebinloeverlabboiftssywshteenmicadmeixnpiosstuerreedofasT-6a24s6inwgalse eivnatlruaavteendousin imnajleectiaonnd fatemalleevels oafny5,of10t,he 1a00n,imaalnsd f50o0llmogw/ikngg. admTihenriestwrearteionnooftestthismamtaetreiraila-lr.elated effects in 000323 Steven M. Glaza Study Director Acute Toxicology Page 14 of 38 SIGNATURE HWI 6329-159 Aas Date 000324 Page 15 of 38 PATHOLOGY REPORT HHT 6329-159 Tnehcerroepswye,rethtewosmkaelleetarlabmbuitssclesacsruirfriocuenddinogn tDhaey l1 uomrbarDayspi2naalndconercdroipnsiAendi.mal At NNoo.. FF5544100766 ((510000 mngg//kkgg ooff bbooddyy wweeiigghhtt)), watshedisfkfeluesteally mduasrcklereds.urroIunndAniinmgalboth htihnedseTiannbismalasndartehe isnadcircaaltivsepinoafl icnojrudrywaosf duinfdfeutseerlmyineddarketireodl.ogyThteo fthiendicnaugdsalin rceogliloencteodf tasherebaqcuki.redThbey lpriovteorc,ol.bile,Aftaendr bneoctrhopksiyd,neytshefraonmimatlhsesweeraenimals were discarded Fo Cor TPhaotthioalsogiE." st Palmer (605312299-5159. 1m) F-/4- FS mate 000325 Page 16 of 38 Sex. Group Male Female Individual Table Body 1 Weights Animal Number Riaznadtoino-n Day -7 Initial {Day 1) 1 (Control) - Sterile Water for FF5544100755 22,,650331 F54081 F54107 2,373 2,544 2,761 2,539 2,526 2,731 FF5544018064 22,,560665 F54074 F54085 2,540 2,553 22,,869302 2,703 2,667 HAT 6329-159 (g) Ternina Day15 Day28 Injection (0 mg/kg 2,933 2,703 - - 2,874 3,086 3,2,173705 -- - 2,99 3,014 Group 2 - T-6246 (5 ma/ka Male FF5544018030 22,,368715 22,,580689 23,,503380 -- F54077 F54004 2,459 2,466 2,592 2,601 - 2,859 2,916 Female F54091 F54108 2,717 2,412 2,785 2,622 2,99 2,790 ~ F54102 F58007 2,690 2,439 2,839 2,533 - 3,29 2,928 Group 3 - T-6246 (10 ma/ka Male F54071 2,487 F54003 2,652 2,2,572736 22,,979039 - F54005 Fs4l0l 2,662 2,622 2,2,674722 -- 33,,116226 Female F54110 F54096 2,719 2,772 2,802 2,959 2,914 3,022 - FF5544009708 22,,574120 22,,878420 - 3,022 3208 - Not required. 000326 . Sex Male Female NAnuimmbaelr F54088 F5544018096 F5344009897 F5544009820 F5a4l1l0s9 Page 17 of 38 IndivTiadbulael 1 Bo(dCyonWteiinguehdt)s (g) HHI 6329-159 Riaznadtoimo-n (Day -7; IDnaiytia1l Terminal Davis paves Group 4 - T-6246 (100 maka! 2,494 2,582 2,771 - 22,,569777 22,,686323 2,803 - - 2,911 22,,356708 22,,676654 3,0-08 3,0-60 22,,440908 22,,654202 22,,686375 -- 22,,570726 22,,963537 -- 32,,299769 Group 5 - T-6246 (500 ma/kg Male 5F544101736 22,,666575 22,,677538 2,8-9 -- FF5S40O82 22,,754374 2,2,788843 -- 33,,222186 Feliz 2,99 3,276 3,530 - Female FF5544008068 22,,556150 22,,667090 22,,88251 -- F5a4l0l7s9 22,,444628 22,,562161 -- 23,,905092 Not required. a pAnriomgarlamNof.orFu5s4e106inwatsheorsitgudiynalalndy wsaesletcrteeadtebdy. theThirsandaonimmiazlatiwoans Nsoa.criFf54i0c9e9d. on Day 2 due to 2 broken back and was replaced with () wNeuimgbhetr wianssutapkeerns.cripted parentheses indicates the day the body b ApnriomgarlamNof.orF5u4se076inwatsheorsitugdiynalalnyd wsaeslecttreedatebdy. theThirsanadnoimmiazlatiwoans Nsoa.criFfs4ilcle2d. on Day 1 due to a broken back and was replaced with 000327 Sex Male Female Male Female Animal Mumber Group 1 F54105 F54075 F54081 FS54107 F54104 F54086 F54074 F54085 F54083 FS4100 FS4077 54094 F54091 F54108 FS4102 F54097 Page 18 of 38 IndividualTabClleini2cal Signs HHI 6329-159 Hour (Dav1) __ Day _Observation~~ 0.5 2 4 2 3tols (Control) - Sterile Water for Injection (0 ma/ka Appeared normal J 4 J J J Appeared normal J J 4 4 4 Appeared normal J J J J v Appeared normal J J J J , Appeared normal 4 J J J J Appeared normal J J J J v Appeared normal J J J J v Appeared normal / / J J v Group 2 - T-6246 (5 ma/ka Appeared normal J 4 4 J v Appeared normal J J J J v Appeared normal J 4 J Js pppeared normal J J 4 4 4 Appeared normal J 4 J J 4 Appeared normal J J J J v Appeared normal J 4 J J v Appeared normal J J J J v 16to28 * * v v * * v v * H / v * * v v *7 ACnoinmdai]tiosnacreixifsitceedd. on Day 15. 000328 Page 19 of 38 Table 2 (Continued) Individual Clinical Signs HWI 6329-159 Sex ANnuimmbaelr Observation 0Ho.u5r 2(Day 41 2 3folsay 15178 Group 3 - T6246 (10 ma/kg Male F54071 Appeared normal v7 7J J * F54093 Appeared normal 7v77 v/ * F54095 Appeared normal 7 7 J v/ 7 7 F54101 Appeared normal vJ4 v v 7 Female F54110 Appeared normal vJ v 7 / * F54096 Appeared normal 7J J 7 7 * F54090 Appeared normal JJ 7 7 J J F54078 Appeared normal J7 v v J v Group 4 - T-6246 (100 ma/kg Male F54088 Appeared normal v7v7 v * F54106 Appeared normal Jv J v - Broken back - - - v Moribund sacrifice - - - 7 F54089 Appeared normal 77JJ J J F54087 Appeared normal 777J v v F54099* Appeared normal '7JJ J * Female F54080 Appeared normal 74 7v J * F54092 Appeared normal +7 vv J * F54109 Appeared normal 7 7 Vv Vv Vv v F54114 Appeared normal vvJJ 7 v * Animal sacrificed on Day 15. /~ CCoonnddiittiioonn enoxtisteevdi.dent. 2 Replacement animal for Animal No. F54106. 000329 Page 20 of 38 IndiTvaibdlueal2 C(lCionnitcianluedS)igns HHI 6329-159 Sex Animal Number _ Observation Hour 05 (Day 2 4 Z Day 3tols 6te Group 5 - T-6246 (500 ma/kq Male F54076 BArpopkeeanredbacnkormal J4J St Moribund sacrifice - - F54113 Appeared normal J J J J / * F54082 Appeared normal J J J J v v FS4111 Appeared normal J J J 4 v v F54112 Appeared normal J J J J v * Female F54098 Appeared normal J J J 4 v * F54084 Appeared normal J J J J v * F54079 Appeared normal J J J J v v F54116 Appeared normal J J J J v v * / ACnoinmdailtiosnacreixfisitceedd. on Day 15. t- CCoonnddiittiioonn not evident. evident at time of 12-hour bleeding interval a Replacement animal for Animal No. F54076. . 000330 ANnuimmbaelr 54106 F54076 Page 21 of 38 Individual TPaabtlheolo3gy Comments HHT 6329-159 Sex --DT i--ed eSs aDcariyft _ic_e_d Necropsy Observation Group 4 - T-6246 (100 ma/ka "- 2 Tlhuembasrkelseptianlal mucosrcdleissudrirfofuunsdeilnyg the dark red. Group 5 - 1-6246 (500 ma/ka: "- 1 Tbhoeth shkielnedta1limbmsuscalned ssaucrrraolundsipnignal cord is diffusely dark red. 000331 Page 22 of 38 HWI 6329-159 Table 4 Individual Animal Tissue Weights and Bile Volumes Sex AMnuimmbaelr _SDacarifyi= ce iveWeright(gqKidnevs VoluBmiele (mi Group 1 (Control) - Sterile Water for Injection (0 mg/kg. Male F54105 15 86.35 14.92 0.6 F54075 15 60.25 15.97 0.1 F54081 28 80.597 14.764 1.2 Fs4107 28 80.649 15.620 0.4 Female F54104 15 98.40 16.03 0.7 F54086 15 77.30 15.23 0.7 F54074 28 82.895 18.793 1.6 F54085 28 80.950 14.925 1.3 Group 2 - T-6246(5 ma/kq Male F54083 15 73.15 14.87 0.7 F54100 15 77.90 17.62 0.7 F54077 28 75.699 17.864 0.3 F54094 28 78.689 17.213 0.8 Female F54091 15 84.90 14.92 1.6 F54108 15 79.90 15.40 0.4 F54102 28 101.494 18.308 17 F54097 28 73.621 16.214 1.4 000332 Sex Male Female Male Female Page 23 of 38 HHI 6329-159 Individual AnnetTiseus Neldhes. nd Bile Values ANnuimmbaelr SacDraiyfice _iW __ve erig(Kgqih )dnet vs GrT oup36 - 24 (10na6 /ke) VoluBmiele (ml. F54071 15 77.10 15.88 0.5 F54093 15 81.40 15.50 0.8 F54095 28 85.504 15.561 1.2 F54101 28 85.668 16.120 1.0 F54110 15 83.68 15.79 1.4 F54096 15 78.10 18.70 1.1 F54090 28 76.220 15.878 2.1 F54078 28 83.460 18.130 1.0 Group 4 - T-6246 (100 ma/ka F54088 15 84.26 16.98 0.4 F54106 2 82.301 16.550 2.0 F54089 28 71.796 16.434 1.0 F54087 28 83.710 18.591 0.6 F54099 15 82.143 16.038 0.5 F54080 15 75.42 18.32 1.2 F54092 15 78.36 16.68 * F54109 28 73.958 16.915 1.6 F54114 28 80.966 19.404 2.0 2 Replacement animal for Animal No. F54106. * This animal had no gallbladder, so no bile was collected. 000333 Sex Male Female Page 24 of 38 HHI 6329-159 Individual Amina]Tiseul Reisheeond Bile Voluses MANnaimmbaelr Day Sacrifice LivWeeright K(gidweys VoluBmiel(enl) GrT oup56 - 2(4 500ma6 /ka) F54076 1 78.058 14.695 2.0 F54113 15 91.52 16.57 Lo F54082 28 88.965 19.431 i.6 F54111 28 83.823 19.405 1.5 F54112* 15 97.824 21.069 1.4 F54098 15 78.95 14.72 1.2 F54084 15 66.42 13.18 0.8 F54079 28 83.117 14.785 1.8 F54116 28 71.962 15.793 2.0 2 Replacement animal for Animal No. F54076. 000334 Page 25 of 38 APPENDIX A Protocol Deviations Protocol TP8084.PK Protocol Amendment No. 1 HWI 6329-159 000335 Page 26 of 38 Protocol Deviations HHT 6329-159 Protocol CP.ageDos6,ing7. PrEoxcpeedruirmeesn,tal(1)Design, DionsjiencgtioRnoutien.to tIhnetrmaavregnionuasl ear vaepipnroxoifmattheelyrig3h0t toear60 ovseerconds. Actual Procedure TGhreouprate feomfalie"nje(cNot.ionF54f0o9r8)one wtaisme 22forsedcoonsdes.admAilnsios,tratthieon Nfoo.r Fon5e408m7a)lewaasnimianlcor(rGercotulpy4, druercaotridoend otfhusdostihengexcaanctnot be determined. Touhtecsoemedevoifattihoensstuadrye. not considered to have had an adverse effect on the 000336  WISCONSIN : RREISRNNG AR Page 27 of 38 + CORNINGCanons Sponsor: wo St. Paul, Minnesota PROTOCOL TP8084. EXT Stuy Title: Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits Date: March 30, 1995 Perforning Laboratory: MHaa3d3zi0ls1eotnoK,ninWsWinissaccnoonnBssoiiunnl,ev5aI3rn7dc04 aboratory Project Identification: HWI 6329-159 } 000337 PErVoRnEeSS sMoAsIL241OEsIaVnEsAT 11 KimswAN 8Y0 MADISFOaNx. sowinneias7i2m2 Page 28 of 38 3 TPP8ag0e842. EXT STUDY IDENTIFICATION Single-Dose IonftrTa-v6e2n4o6usinPhRaarbmbaictoskinetic Study HUT No. 6329-159 Test Material T-6246 Sponsor Sponsors Representative : TMoxicology Service 3MMCeednictaelr, DeB8p1adrg.tme2n0t-26-02 SPt..0. PaBuolx, 3M3N22055133-3220 MJonhn L. Butenhoff, PhD ToMxeidcioclaolgyDeSpearrvtimceent 3P.M0.CenBtoexr,332B2l0dg. 220-2-02 S(t6.12)Pau7l3,3-1MN96255133-3220 Study Director HSatzelveetnonH.WiGslcaoznasin, Inc. MPa.0d.isoBno,x W7I54553707-7545 (608) 241-7292 Study Location H33a0z1letKoinnsmWainscoBnosuilne,varIdnc. Madison, WI 53704 ProEpxopseerdimeSnttuadly STtiamrettabDlaete DErxapfetrimReenptoarlt DTaetremination Date April 4, 1.995 MJuanye 1,12,19915995 000338 Page 29 of 38 TPPaag0e8s3. EXT 1. Study Single-Dose Intravenous Pharmacokinetic Study in Rabbits 2. Purpose To assess the level of systemic exposure when the test material is administered as a single intravenous injection to rabbits 3. RegulatoryCompliance . This study will be conducted in accordance with the following Good eLxacbeoprtaitoonrythPartactaincaelysRiesguolfattihoenst/eSsttanmdaatredrsi/aGluidmeilxitunreesswiftohr the conducted concentration, solubility, homogeneity, and stability will not be [[x1] 2C1onCdFuRct58as (aFOAN)onregulated Study [] 40 CFR 160 (EPA-FIFRA) [ ] 40 CFR 792 (EPA-TSCA) [ ] c(81)30 (Final) (OECD) [] 59 Nohsan No. 3850 (Japanese MAFF) [J Notification No. 313 (JapaneseMOH) work. Al] procedures in this protocol are in compliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study does not unnecessarily duplicate any previous 4. Quality Assurance The the protocol, study conduct, and the final report will be audited Quality Assurance Unit in accordance with HazletonMisconsin by (HWI) Standard Operating Procedures (SOPs) andpolicies. 5. TestMaterial A T-I5de2n4t5ification 8. Physical Description (To be documented in the raw data) C. Purity and Stability dTteehsetterSmmpiaontnaestorriioanlas/svseu(hmiiencsclleurdeimsnipgxotnusurinebd(ieslr)ittyfeosrtfoccroonnpdcuiertniittorynasta)in.odn,stSaasbmoiplllueibstilyiotfy, homogeneity, and stability analyses will be taken before administration if requested by the Sponsor. These samples (if ttaekremni)natwiiolln. be sent to the Sponsor after experimental ! D. SRtooormagteemperature 000339 Page 30 of 38 } TPP8a0g8e4.4 EXT E. ReRseseerrvvee Ssaammppllee(ss) materials will be otafkeenachforbattchhis/losttudoyf. test and control THhIe for tiensta afnrdeezceonrtrsoelt 10 years per HI mtaotemraiianltairnesearvteempsearmaplteusrewiolfl -b2e0Cstor1e0dCat SOP. The, Sponsor will be contacted after r1e0gyuelaartsoryforGooddispLoasbiotriaotnoryinPraaccctoircdeasn.ce with the appropriate F. RAneytenutniuosend completion tofestthmeatienr-iialfewiplhlasebeofretthuernesdtudtyo. the Sponsor after 6. ASsaferteyquiPrreedcabuytioHHnIs SOPs and policies 6. Control Materia A. SItdeernitliefiwcaatteironfor injection b B. PhClyesairc,alcDoelosrclreispstilioqnuid C. PTuheritpyuriatndy considered Santdabisltiatbiylity of adequate for the ptuhrisposUeSsP gorfadtehismatsetruidayl. is 0. ReSftroiraggeerated . E. RSeeeserSveectiSoanmpl5.esE. Reserve Samples F. ARanneydtenrwitelimloaninniotngbecondtirsoclardmeadteraifatlermaiyssbueancuesedoffotrheotfihnearl treesptoirntg. G. SAasferteyquiPrreedcaubytionWIs SOPs and policies 7. Experimental Desian A. Animals (1) SRapbebciites ) (2) HSrtar:ai(nN/ZS)oSuPrFc/eHRP, Inc. 000340 Page 31 of 38 y (3) AsIAednutaitttiation TPP8ag0e845. EXT (4) 2Mei5ahtt 3at5kInitiation (5) N20ummbaelresandandSex20 females (6) Idnednitviidfuiaclatinounmbered ear tag (7) Husbandry (2) HIonudsiivnigdually, (heavy gauge] in screen-bottom stainless steel cages (6) FAoomdeasured (PHI Feeds, amIonuc.n)t. ofThLeaboforoadtor1y3 RraobubtiitnelDyietanHaFlyz#e5d326by tehnevirmoannmuefnatcatlurecrontfaomrinnaunttrsi.tional components and ! (c) 4wadattee1rribairteunanfarloymzeadn bayutoWWmIatifcorstyostteaml. disSsaomlpvleeds osoflitdhse, hhseypaedvcryiofcimaeerdtbaolmnsis,croobrigoalnoogpihcoaslphactoenst,enta,nd scehlleocrtiendateeldements, (4) CTthoheantrteawmoiaunrlaedntnsoinktneorwfnerecownittahmintahnitssstiundyt.he food or water (2) EEsnentvviirrtooonnmmmeaenintntatlainconatrtoelmsperfaotrurteheofani1m9alC troaom23wCi,ll abe rTeilgahtti/v1e2-hhouunridditayrkocfyc5l0e%.520%, and a 12-hour (7) AAcEclTiemaasttio7ndays (8) SBaadeseleqeducattioeonnhneuofamTbteThrestaofndAnexibtomrdaaylswaeniigmhaltsawciclolrdibnegputorcWhWaIsedSOPsso. thAant )) a7s0ntirmaaatnliinsfailewdililnboodbybeviwpeoliuagschletyd rpioanontrodomhsietzauladttyhiognriosupprpsolagcrueasdimngwointahtienst.nineThe days of study initiation 000341 Page 32 of 38 ) PPa8g0e84.6 EXT (9) JHiussttoirfiiccaaltliyo,n the aninal of tfhoer SNpeewciZeesalaSnedleWchtiitoen choice because of the alalrbgienoamroaubnbtitofhas been background information on this species. 8. Dose Adninistration (1) TestGroups Group. TesMta/tCeornitarlol Dosmeo/kLee)vel_ MNaleus omfAbFneimemaallerss 21 ((LCoown)trol) SteTr-i6le24w6ater 05 3 (Hedium) T-6246 10 44 4 44 4 54 ((HHiigghh)) TT--66224465 150000 44 44 ab TTwhoe The daroensmieamianlviosnl/gusneexa/nwdiimolaslles lbee(vte0wl.o5 waninlLil/nkagbleso/sfseaxcb/orddiyofsiecweedilgehvoten.l)Daywil1l5. be sacrificed on Day 28. ' C. DosingProcedures (1) rDIoinsgtihrntagveenRaooruustoveeir'njeacptpiroonximianttoelyth3e0matrogi6n0alseceoanrdsv.ein of the (2) RIenatsroanvenfoours DoisnijnegctiRoonute1s systenic exposure. an acceptable route to assess (3) DSoisnignlegDduorsaetion (4) DTohesedaPyrepoafrattrieoantment Group 1 animals will wbielltrebaeteddeswiigntahtesdterasileDaywat1.er Tahtea ddoisleutevdolwuittehofst0e.r5ileml/wkagt.er Tthoeatcehsitevemataersipaelciwfiilcl be cIonndcievindturaaltiodnosefsorwielalchbdeoscealcTeuvlealtedinbaGsreodupson2-t5h.e animal's abdomdiyniwsetirgahttiont.akenTjhuestprebpeafroerde tteesstt mmaitxetruiraels at room temperature until administration. will be stored J 000342 Page 33 of 38 i 0. ObservoafAtniimoalns PTaPg8e0847 EXT (1) Clinical Observations The animals will be observed for clinical signs of toxicity before at approximately test 0.5, or control 2.0, 4.0 material administration, hours post-injection ds(aaDicalryyif1i)(,ca.ema.nidntaednrdavialply.m.t()hDaeyrfoera1f5mtooerrrtaDflaoyirtyc28l)iu.nntiiclaOlbstehsreivgsanctshieoadnnusdlemtdawyice (2) Body Weiants be extended when directed by the Study Director. injection (Day 1). at the scheduled sacrifice mterval For randomization, before test or control material (Day 15 or Day 28), and at unscheduled death and sacrifices (when survival exceeds 1 day) (3) Sample Collections (3) Frequency 4-, 8-, 12-, 24-, and 48-hours post-injection, on Days 8, 15, 22, and at the scheduled sacrifice interval (Day 15 or Day 28) (b) Meithood ofnCol/lecNt unb of Aenimrals Blood samples (approximately 4 nL) will be collected from the marginal ear vein (left ear) of all animals at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8. Additional samples will be collected on sacrifice on Day 78. Days 15 and 22 for the animals scheduled for Approximately 20 mL of blood (actual volume to be dpoocsutmeerniotredveinna thceavarawatdatthae) twiimlel ofbe thoebtasicnheeddulferdom the sacrifice (Day 15 or Day 28). Approximately 20 mL of blood will be collected from moribund animals during the study, also, if possible. The samples will be stored at room temperature and then centrifuged, and the separate serum and cellular fractions stored in a freezer set to maintain a ctteheelmlpueSlrpaaotnrusroferracoatffitoe-nr2s0eCwxiple+lr1i0mbCee.ntsaelnTthetefrrsmoeigpneaanrtaitooennd.drsyerTuhimeceantdo dSipsopnossoirtiiosnreofspotnhesibslaemplfeosr. the retention and ) 000343 Page 34 of 38 ) PTaPgae0848 EXT Samples will be shipped to: JMamEesE0. BJPohCnson B9l3d5g.Bus2h-3A-v0e9nue St. Paul, HN. 55106 JtaenleesphoD.neJoathns(o6n12)or77hi8s-52a9l4terpnraitoer the samples. wtiollthebeshnioptmiefnitedofby E. Termination (1) AUnnyschaenidmualleddySiancgrifdiucreisngatndheDesattuhdsy or moribund condition will be subjected stoacranifiacbebdreviniataed grarenocesossrtdheendte.iczreodApnswiyimtaehlxsamsioinndaituainomnopreinatbnoudbnadarlblciotnaadblintoir(omvnaialiwitilinlejsebcwetiiloln be in e7t.hxEes.anmga(ur3ig)nianStaaeldm.pleearCTovilesliseunec)s,t,iobnla,sedwdievlsilcaribtbeheedcvoelinlnaecstceeacvdtai,ofnroamndany caonnimdailtiodny.ing"Adfutreirngnectrhoepssyt,udytheoransiamcarlisficwieldl inbeadimsocrairbduendd. (2) OSncheDdayuleIdS, Sathceriffiicrsets two animals/sex Tevel (based on the group assignment rasasnidgonmeidzattioone)achwildlose bine tahneesmtahregtiinzaeld exsanguinated. weiatrhvesiond)i,unblpeedntovbiaarbtihtealven(aviacavian,jeacntdion The remaining two animals/sex/dose level wciianlvjlae,ctbieaonndaneeisnxtshatehnetgiuzimenadragtiewndiatlhonesaDoradyivuem28i.np)e,nAtnobblaeardbbbirvtieaavliatth(eevdiavgernoass n(eascrdoepsscyriebxeadminiantisoenctiwoilnl 7.nEo.t be collected b(e3) doSnaem,plehowCeovlelre,ctitoins)suewisll (3) TshaemplweholCeollTeivcetri,onbile, and both kidneys from each animal 2bwiilltlee)m,pbeeraacntdoulrlieemcmtoeefddi,a-t2ew0leyCighp1el0dacCe.(dvoliuAnmfetaerofnrlesyeazmepdlreetescertomlilnteeocdtmiafooirnn.,tain the animals will be discarded TTohnheedrssyaammppillceeesstoa(n1ditvheetrh,eSiprobinlsceoo,rrreaasnfdptoenrkdiidennxgepyesrw)iemiegwnhitltlasl obertevrsomelniutnmaetfsi.roonzen J w7ain:ld0l3d)ib,espos0sh0i)i,tpipoendThetofoSpttohhneesaspraemrpsilsoensr.elsipsotnesdibilneSefcortiotnhe retention 000344 Page 35 of 38 \ PPa8g0e883. EXT FL Statistical Analuses No statistical analyses are required. 8. Report A final report including those items listed below will be submitted. PDDreeosscccerrdiiupprtteiisoonn ooff tthhee tteesstt sanydstceomntrol materials Dates of experimental initiation and termination Description of any toxic effects Gross pathology findings (if applicable) Gross pathology report (if applicable and requested by theStudy Director) 9. Location of Raw Data. Records, and Final Report Individual animal tissue weights and bile volumes Original data, or copies thereof, will be available at HWI to facilitate auditing the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data, including those item listed below will be retained in the archives of HWI according to HWI SOP. , DPIornso-teToicfpoerlerpeaacrnoadrtdipsornotorceoclordasmendments BDOboosdseyervwaaedtinigiohnntissstration SSahmipplpeingcolrleeccotrdison records PSattuhdoylocgoyrreRsecpoornddsence Final report (original signed copy) Tnhoet bfeolalorwcihnigvedsuwpiptohrtitnhge rsetcuodrydsdatwai.ll be retained at HWI but will Animal receipt/acclimation records Water analysis records Animal room temperature and humidity records Refrigerator and freezer temperature records Instrument calibration and maintenance records J 000345 Page 36 of 38 |. PPa8g0e841.0EXT PROTOCOL APPROVAL ohFn eL. B2uteKnnhiofl,oiPLRDaff MSponsors Representative Steven H. Glaza . ASctuutdey TDiorxeicctoolrogy , Hazleton Wisconsin, Inc. RQeuparleisteyntAastsiuvreance Unit Hazleton Wisconsin, Inc. (6329-159.protdskz) Date Y-9 Date 330.9% 3-30-95 ate ) 000346 @ HAZLETON Yo LSelSNISN Page 37 of 38 +commer PROTOCOL TP8084.EXT Single-Dose IntrTa6v2en4o6usinPRhaabrbmiactoskinetic Study of HHI 6329-159 Sponsor 3MMeTdoixciacloloDgeyparStenrevnitce 37.M0.CenBtoexr,332B2ld0g. 220-2-02 St. Paul, MN 55133-3220 Soonsor's Representative Contractor H3a30z1letKoinnsnWainscoBnosuilne,varIdnc. Madison, WI 53704 Study Director -_John L. Butenhoff, PhD ) nnn Steven M. Glaza Anendnent No. 1 This amendment modifies the following portions of the protocol: I. Page 6. 7. Experisental Desian; B. Dose Adinistration: (1) Test Groups Effective April 4, 1995 The test material mixture for Group 5 could not be prepared at the concentration needed to utilize a dose volume of 0.5 mL/kg. Modify footnote "a" in this section with the following underlined addition: a Th2e% adnodse0.v7o5lummel/kwiqllforbeG0r.ow5smL5./kg of body weight for Groups J 000347 Page 38 of 38 NV Amendment No. 1 + HWI 6329-159 Page 2 2. Page 6. 7. Experinenta)EffDeecstiiavnes A6.priDlos5e, A1d9m9i5nistration: (1) Test Groups. Animal No. F54076 (Group 5 male) and Animal No. F54106 (Group 4 male) were sacrificed on Days 1 and 2, respectively, due to injuries (apparent broken backs). Add the following paragraph to this section: D(uAenimtaol thNoe. sFa5c4r0i7f6i)ceaonnd oDnayDa1 yo2f oonfeonGerouGprou5pma4lemale (Animal No. F54106) because of injuries (apparent broken backs), rseapmelamcaenmneenrt aasnimfaorlstwhiellinbietiatlreaatneidmalast itnhethseamestuddoys.e lTehveels in the observations (clinical observations, body weights and sample collections) and the termination of the animals (unscheduled sacrifices and deaths, scheduled sacrifices, and sample collection) will be conducted fn the same manner as for the other animals in the study terminating on Day 15. PROTOCOL AMENDMENT APPROVAL Plo 2. BiB SJophonnsLo.rsButReenphrofefs,enPhtDvate 3M Toxicology Service Medical Department Fes Date SSttuedvyenDMi.reGcltaozra Acute Toxicology Hazleton Wisconsin, Inc 4-185 Date | QHRuaeanplrieettsyaennAtshaistsiierjoaemnscien,UniTtne (6329-159. Aml.dsk3) ate 4.18 9 000348 Calibration curve of 95050481 Fluoride L ---- EER 5I RrnenE aEene Tr nr rea ge | 21851733 { Al | Ad : ; | So I 1||1 // /7 : Iii I 1 / I 3 / H A Hl fl / I 5)1 7/ : ib. i gi Hy 4] JS i 1 py || 4 /# I 11 | / 1 1| 7 iH I TA Lo 0.001438 Tomer a Order 2 |i i pe Heasuree d red500i r: o.seee } 000349 000169 2 -------------- Calibraticn curve of S50504A1 Fluoride $.5 -------------- HERTEES O YIN N GE T | .7141803 A i / I !1 Ww : { 1| // / i|| | / i i| 3! /A{/ iiPil iii |Hl 41 ol // Ii Hi / | / I i he // ii | J i | Sf IN p/ iIi 1 i| Ee | 1 ii Lo ot 1fi 4.0563468 =---------------------= | a Measured 40es | Order: Inverse Logariths s 8174 | 000350 000170 28 Raw data of 3505041 : Fluoride 1.5 ------------ TTuE asTT TNT s SERRE | 4085 Ti T[a d RE Ld wl Po P Lhla 1Lhil 1 Pi ah Ju Hl Lh im A in Leiai bHlH Hi 0g I Hl 1 i | bi o bSoEl. od| i ho h| oPi Hil Hilt i. HH] TRiIE iH | Hi li oii vod Ei H HPhh { Hi 1 TiHli 2 {I flaith T-eallii 1: Hid s i atl i Til 4 dE ts 1 | DCofeUis BR L 4 PhEI U Tid weRe wA AiAtaTeR a B TE eh LEANER TE Te IN VY 0| | T yn epee | o Tine 000| so=Exit | Fishelp | Crti-P=Edit peaks | 000351 000171 2