Document gb3n8gVzMrMnm9EeQ7MpoJQNe

. AR22G-0283 T1N-6E1I5FOo,Su5Et7zB3inl0se 3M MEDICAL DEPARTMENT, CORPORATETOXICOLOGY Protocol for Study No. T-6316.15; ST-30 N-EFOSEBileMethodDevelopmentin Rats Study Objective: `The objectiveofthis EtFOSE treated rats study is to for method collect urine and bile `specimens from control and Ndevelopment. This study will support the N-EtFOSE ADME studyin Cynomologusmonkeys currentlyunderwayatCovance Madison. Research Client: 3M Specialty Chemicals Division Biodegradable Surfactants Team 3M Center, Building 236 Saint Paul, MN 55133-3220 Sponsor: 3M Specialty Chemicals Division 3M Center, Building 236 Saint Paul, MN 55133-3220 Study Location: 3M Strategic ToxicologyLaboratory 3M Center, Building 270-35-06 room SB314 Saint Paul, MN 55133-3220 Study Director: Andrew M. Seacat, Ph.D. Sr. 3M MReedsiecaarlchDeTpotx.ic/oCloogripsotrate Toxicology 3M Center, Building 220-2E-02 Saint Paul, MN 55133-3220 Ph. 651-575-3161 FAX:651-733-1773 Study Taxicologist: Deanna Luebker, MS A3dMvaMnecdiecdaRleDseepatr.ch/ CToorxpicoorlaotgeisTtoxicology 3M Center, Building 220-2E-02 Saint Paul, MN 55133-3220 Ph: 651-737-1374 FAX: 651-733-1 773 ProposedStudy Timeline: In-Life Start Date: 8/12/1999 In-Life End Date: 8/20/1999 TRheigsusltautdoyrwyiClolbmpelpiearnfcoer:med inthe 3M Strategi|cToxicologyLaboratory under adefined pTrooxtioccoolloagnydLcalabssGiLfiPedParsogar"aCmlaPsrsocBeSdutrued.y"asexplained in TOXSOP0950,Strategic 004336 ! T4316.15o;a/S1T33109 N-EWFOSE Bile Test Material: Dan Hakes, Product purity N-EtFOSE. Responsibility Liaison 3M Chemicals Division, will furnish high IdentifiNcaamtieo:n:Narrow Range N-Ethy! perfluorooctanesulfonamido ethanol, FMMolecul3a92r3F.ormula: CSF17S02N(CH2CH3)CH2CH20H Lot Number: Lots 30035, 30037, 30039mixedandanalyzed as one sample. Purity: 98.14% Stability: Storage`DCoocnudmietnitoantsi:on will be kept on ile. Utpeompnerreacteuirpet.,test material willbestoredtightlysealedat room Characteristics: Idenffionrematthieotnesotnmastyenrtihaelsiwsilmletbheokdesp,tcoonmpfoles.itionorothercharacteristics that Animals: SSptercaiiens:: RSaptrague Dawley Source: Harlan ASpgeeciafticiantiitioantsi:oBnoilfetCraenantumleantte:d --Fre6e-8Enwdeeks `Weight at initiationoftreatment: approximately 150-250g Number and sex: 20 males, 20 females Table 1- Dose Group [Grp-- Those Bo Tomgkg TN 1s Tse mae | PB T Tomeo kg m [I5S gIMFk emmeie e|| BO Toomgie 15 [Femse J Identification: AUA Number: eartag withanimal numberorunique tailmark. 2246 Husbandry: HousinUgp:onarrivalat 3Mallanimalswilbeindividuallyhousedinstandard tcraagness.feArrleld rtaotsifnrdiovmidguraolumpesta2boalnidsm4caangdes1efaoclhlfowriongmgdroosuinpgsa1nadnhdou3sweidllbe 004337 2 T3N-1E6U5F0O,8S/ES1T3B/i39l0e9 hthoeurseetdhirnoustgahnddaaryd4-cpaogsets dtohsreo.ugAhloluotthtehrersatusdyw.ill remain individually Diet/WaHtaerrl:an Teklad LM-485 Mouse/Rat Sterilizable Diet, supplied by Harlan `Teklad, Madison, WI, throughout the study. and tapwaterwill be provided to al raad ltibistum EnvirEonnvmiernotn:mentalcontrols fortheanimal roomwillbeset tomaintain a tofemrpoeormaatiurrepeorf7ho2u+ranFd, ahu1m2ihdoiutryloifgh3t0/-d7a0rk%c,ycalme.inimum of 10 exchanges DoseanMdeDtohsoidnogfPardomciendiusrtersa:tion/Dose preparation: toAsriantgslieng2r0omugp/skg2daonsde o4fonNd-EatyzFeOrSoEowfitlhlebsetuaddmyi.rTihsteedroesdivinagsoorlaultgiaonvawiglel b1e5pmrieptaisrseudeagrsin2d0e.r0.4%Av(o4lmgu/mmeio)fun5imfiorsmussopleuntsiioonni/kn2g%bodTyweweenig8h0tuwsiilnlgbea astdromkeisonftihsetotteiesascurehegrraditn.deRr peestel-befsoreoeufascoshlisdaspmwpilleelbiesnpderrasfwon-riumpeidonwtihtneh5 wsilylberafdiomrindnoisgsitneegr.edAvsiiangorlael g5amvlag/kegtobroadtysiweniggrhotudposs1eoanfd23%oTnwdeaeynze8r0o ofthe study (see table 2). ObservationofAnimas: ClinicEaal cObhsaenrivamtailownisl:lbeobserveddailyformortalityandmorbidityandnotable finwidllbiernecogrdesd. Additionalfindingswillberecordedas theyare observed. Body EWeaicghhtasn:imalwillbeweighedimmediatelypriortodosingand immediately prior to euthanasia Specimen ColleFcrteiqoun:ency (See table 2): doBislienwgilwliblecsoelrlveectasedcodnatirlyolubpiolen.)arrUirvianleatco3llMe.ct(iAolnlsbwiillelcboellmecatdeed prior to on days 1 -4 postdose. Necropsieswillbeperformedonday4-postdose. 00433g 5 : Tot615ou:i5s7n3s Ta 2 b - Scl hedue le N-E{FOSE Bile [Sa] Mon] Tus] Wed] Tuugm] AFwdl] Amesnatr] BAireacFoi@.| Biles| Bitsoe Rbepal] Bicol| Ar"iBoel BecDoO.W|| Am"iB]a Bpiocnool|| e"gBa Bpiocdoel|| Aiuas] Bpaisetcieosn]| Reugmi Bhpeoesaens Uroinn]e| Uroinn]e| Ueare|| Uri hoen,m Method of Specimen Collection: Bile willbecollected fromeachratdaily upon arrivalat 3M.The exteriorized portionof thecatheter willberemovedfromthe pouchofthe protective jacket andthewire plugintheFreeEndcatheterwillbe removed. Anewly opened syringe wibelusl ed to withdrawn bile from the catheterofeach rat. The bilefromeach ratwillbeequally splitand placed into 2labeled tubes. Thewireplugwillbeinserted backintothe catheter ifnoltlhoewpinogucbiholefctolhleecptriootnecatnidvtehjeaecxkteetrtioorpirzeevdenptorctaitohnetoeftr hcheewcaitnhge.teErapclhace `tubeofbile willbe frozen (-70C)untilanalysis. Urine willbecollected fromeachmetabco agleoin s dam ys 1-4 postdose. Foreachurine. `specimen,theinitialvolumewillberecordedand the sides ofthe urine collectionapparatuswillbewashedwith 10-20 mldeionizedwater.The `urineandwashfromeachcagewill be collectedin a 50mltubeandthe finalvolumewillbebroughtto 45 mlwithadditionaldeionizedwater. Each urinespecimenwillbe frozen (-70 C) until analysis. On da4y post doseanimalswillbeeuthanized by CO,andgrossnecropsyperformed. Duringnecropsy,blood (~ 6mi)willbe collectedvia the abdominalaorta and transfetorbrleoodd collectiontubeswithout anticoagulant. Blood tseammppelreastwuirlel,baenadltlhoewceldottowcillotbfeosrpuanpedroiwondoifn1a5cetnotr3i0fmuigneuattes1a10t0roxogmfor 5minutes. `Theserumwillbetransferredtolabeled 1.5ml microfugetubes andcentrifugedagainat 2000 x gtoremoveany remainingredbloodcells. `p Eacho seralsamy pmliecwp riolflutr gheetnuo bbeetrap annds(fy -er7r0el dCt)oue natseipln aanraalte yesilsa.bEealecdh liverwill be removed, rinsedin saline, weighed, placed into a labeled sterilesample bagandfrozen (-70 C)untilanalysis. Specimen Handling and Analysis: S8p0ecCimienntshweiSltlratteemgpiocraTroixliycboelosgtyorLeadbionraatofrrye.ezer set to maintain-60 to - 004339 ` Te316.15,5T 30 08/13/99 For metabolite analysis, urine,liver and N-EtFOSE Bile sera will be packed indry ice and `shipped to: Kris Hansen, Ph.D. 3M Environmental Technologyand Safety Services 935 Bush Avenue St. Paul, MN 55133-3331 Ph: 612-778-6081, FAX: 612-778-6176. For analyticalmethoddevelopment iced and shipped to: purposes, bile will be packed in dry Karsten Levsen NFirkaoulnahiofFeurchIsnssttirtu1t.fDu-er30T6o2x5ikologieundAerosolforschung Hannover GERMANY `The Strategic Toxicology laboratory will retain onehalfof all the bile `specimenscollectedforin-house method development, verificationanalysis `and/or as a retain for possible future analysis. Allresultswillbeprovidedforinclusioninthe finalreport. `The number,type anddateof collectionofspecimenstobe generatedfor analysis are as follows: a Tab3l- e a Specimens -4 =3 =. PF] Ee Fraunhofer El [er0[rT aayz Tay Tooy4 [Tom 2 -1 PD PD PD PD ll ll ll house is [[T T SUaeem T TTTuT | {5|T2#0 |& Jw ve 1 {1 T -1 T20 Ta Data Analysis: Data collectedon N-EtFOSEmetaboliteconcentrations inserum,liver andurine using validated methods willbeanalyzed for toxicokineticparameters andforstatistically significantdifferencesbetween groups using Students T-test and/or ANOVA.Data mceotlalbeoctleidtdeuarnialnygstihswemilelthbeoadnadleyvzeeldofpomrepnrteacnisdivoanliadnadtiaocncpuhraasceysaosfnebicleesasanrdyutroidneevelop a. : validatedmethod. Providedthat enough oftheretained bile andurine samplesare . 004340 s T316.15o;uS1T3-9390 available EtFOSE following the development metabolitelevelsunder the voafvlaildaitdeadtemdetmheotdhsotdothgeeynemraayte then data be anNaNl-yEzteFdOfSoEBriNl-e for usein a. definitivereport Responsibilities: DcoelalencntianLguicn-blkiefersapnecdiAmnednsr,epweSrefaocramtinwigtllhbeenreecsrpoopnssiiebsl,ecfoollredcotsiinngagtnhdesaennidmianlgs, fsoprecmiemtehnosdfdoreanavlyesislanoodrpmveamryipfeiecrafntoirotnm.additionalexperiments onthecollectedtissues KtirsissueHsa,nasnednt,he3uMriEnnevisraomnpmleenstfaoll,lwoiwlilnbgedreevspeolnospimbelentfoofraanvaallysiidsaotefdsmeertuhmoadnfdorluirviernary metabolitesofN-Et FOSEifpossible. rKeasrpsotnesnibLleevfsoernt,hFerdaeuvnheolfoeprmIennsttiotfutafvuaelridTaotxeidkmoelotghioedufnodrbAielrioasroylmfetoarbsoclhiutnegswainldbteh:e + sAunbdsreqeuweSnetaacnaatlyasnidsoDfetahnenraeLtauienbekdebirwleilslamdpralfets,apfrionvalidreepdotrhtearnediesnassuurfeftichieerntespuoprptly. teccivesappropriate 3Mreviewbefore afina reportis issued. 004341 6 : Signatures: TNeeoro08/13/99 --CndDd N Soi vry~zie s fas 7 Dr. Andrew Seacat Date Senior Research Toxicologist Study Director DAdevaannncaeLdueTbokxeirco,liogzist Date Study Toxicologist Selo tr- Tw3lk.2 I1FSOSELRMEE ii) ' 004342 ,