Document 3m3E0rOJGX2dyJRadvxGwwV3

3M MEEDICAL DEPARTMENT, CORPORATE TOXICOLOGY Protocol for Study No. T-7098.1 PRARMACOKINETIC STUDY OF POSF IN RATS 01/07/99 T-7098.1 POSF PK Study Objecdve: The objective of this study isto assess the potentialfor oral absorptioa, urinary and fecal clearanceand biologicalpersistenceof perfluorooetanesulfonylfluoride(POSF) inmale Sprague Dawley ratsaftera singleoraldose. The POSF compound isthe startingmaterial forthe synthesisof a wide varietyof perfluorooetaneslffonate(PFOS) based materials. The purpose of thisstudy isto understand the rateof metabolism of POSF to PFOS by the liver.This studywillprovide data forproper riskcharacterizatioonf POSF. Research Cfient. 3M SpecialtyChemicals Division 3M Center,Building236 SaintPauL MN 55133-3220 Sponsor.- 3M SpecialtyChemicals Division 3M Center,Building236 SaintPad, MN 55133-3220 Study Location: 3M StrategicToxicology Laboratory 3M Center,Building270-3 S-06 room SB314 SaintPauL MN 55133-3220 Study Director. Andrew M. Seacat,Ph.D. Sr. Research To)dcologist 3M Medical Dept. /Corporate Toxicology 3M Center,Building220-2E-02 SaintPauL MN 55133-3220 Ph.:651-575-3161 FAX: 651-733-1773 Study Ta-deologist: Deanna Nabbefeld,MS Advanced Research Toxicologist 3M Medical Dept. / Corporate Toxicology 3M CenterB,ufldin2g20-2E-02 SaintPauL MN 55133-3220 Ph: 651-737-1374 FAX: 651-733-1773 J"IiwposeSdtudy 7"smeline(Assuming EHS&R approval on JarL5'@.): In-LifeStart Date: January IIth,1999 In-LifeEnd Date: February 9th, 1999 AnalyticalCompletion Date: March 22nd, 1999 FinalReport Completion Date: April19th,1999 01/07/99 T-7098.1 POSFPK RegulatOryCOMpli=": Thisstudywillbe performedinthe3M StrategiTcoxicologyLaboratoryundera defined protocoland classifieads a "ClassB Study"as explainedinTOX SOP 0950, Strategic ToxicologyLab GLP Program Procedure. TestMaterial-. Dan Hakes,ProductResponsibiliLtWywn 3M C hemicalsDivisionw,illfin-nishhighpurityPOSF. identirication: Name: PerfluorooctanSeulfonylFluoride Molecular Formula: To be provided. Lot Number: The lotnumberswillbe maintainedintheraw data. Purity: Documentationwillbe keptinon file. Stability: Documentationwillbe kepton file. StorageConditions: Upon receiptt,estmateriawlillbe storedtightlsyealedatroom temperature. Characteristics: hforinationon Wathesismethods,compositionor othercharacteristitchsat definethetestmateriawlillbe kepton file. Animals.- Species: Rat Strain: Sprague Dawley Source: Harlan Age atinitiatioonftreatment: 6-8 weeks Weight atinitiationftreatment:approximately150-250g Number and sex: 30 males Table 1 -Dose Groups Group Dose N Euthanasia *1 0 5 day I postdose *2 0 mgft 5 day4 postdose *3 0 mg/kg 5 day29 postdose 4 5 mp,/kg 5 day 1 postdose 5 5 mgft 5 day 4 post dose 6 5 me/ke .5 day 29 post dose *Rats ingroups 1-3 willbe used concomitantly as conirolanimals in sm&es T-70 71.2, Phwwwolane,ftc Study ofM5.56 in Rats,and T-7099.1,Pharmacolanetic Study OfFX- 845 inRals. Identirication: AUA Number: eartagwitharunw number or umque t" mark. 2154 2 01/07/99 T-7098.1 POSF PK Husbm&y.l Housing: Three specifircatsfrom groups 3 and 6 willbe housed individualilny metabolismcagesforportionsofthestudy(seeTable2). When notin metabolismcages,theseratswillbe group housed instandardcages.AN otherratswillbe group housed instandardcagesthroughoutthestudy. Diet/Water. HarlanTekladLM-485 mouse/Rat StmilizablDeie@ suppliedby Harian Teklad,Madison,WL and taPwater willbe providedto allmts ad libitum throughoutthestudy. Environment: Environmentalcontrolsfortheanimalroomwdl be setto maintaina temperatureof 72 3*F,humidityof 30-700/ca,minimum of 10 exchanges ofroom airperhour and a 12 hour light/darckycle. Dose and Dosing Procedures., Method of administration/Doseprepantion: A single5m&g dose ofPOSF willbe administerevdiaoralgavage to rats ingroups 4-6 on day zeroof thestudy.The POSF willbe preparedas a i% (i mg/mi)uniformsuspensionin2*/oTween 80 usinga 15 ni tissue grinder.A volume of 5 nilsuspension/kg body weightwillbe administeretdo eachrat.Re-suspensionof solidswillbe performedwith5 strokesofthetissuegrinderpestelbeforeeach smple isdrawn-up inthe syringefordosing. A single5 nil/kg body weight dose of 210/Toween 80 willbe administered viaoralgavageto mts ingroups 1-3on day zeroofthestudy. observationofanimals.ClinicalObservations: Each aninw willbe observeddailyformortalitaynd morbidityand notable findingwsillbe recorded.Addifionalfindingwsillbe recordedas theyare observed. Body Weights: Each animalwillbe weighed immediatelypriorto dosing,weejdy thereafter and immediatelypriorto euthanasia. Specimen Coftecdon: Frequency (seeTable2): Urineand fecescollectionwsillbe made on days 1.2,4, 14 and 29 post dose. Necropsieswillbe performedon days 1,4 and 29 postdose. 3 Table 2 - Schedule Jan 10 Jan 11 day 0 DOSING Jan 17 -gu 6 PD Jan 24 day 13 PD Jan 18 day 7 PD Jan 25 day 14 PD Switch to met cages. Jan 31 _gnlo PD Feb 7 day 27 PD Feb 1 _Iffll PD Feb 8 day 28 PD switchto met cages Jan 12 day I PD Collection Dy i PD sac Jan 13 day 2 PD Collection Switchto reg cages- Jan 19 day 8 PD Jan 26 day 15 PD Collection Jan 20 day 9 PD Jan 27 day 16 PD Switch tD reg.cages. Feb 2 day 22 PD Feb 3 day 23 PD Feb 9 day 29 PD collection Dy29PD sac Jan 14 day 3 PD Switch to met cages. Jan 21 day 10 Jan 28 day 17 PD Feb 4 da!v24 PD Jan 15 day 4 PD Collection Switch to reg cages- Dy 4 PD sac. Jan 22 day 11 PD Jan 29 day 18 PD Feb 5 day 25 PD Jan 16 day 5 PD Jan 23 day 12 PD Jan 30 day 19 PD Feb 6 day 26 PD 01/07/99 T-7098.1 POSFPK Method of Specimen Collection: Urine and feces will be collected from each metabolism cage at the designated times. The initiavlolume of urinewillbe recorded, the sidesof the urine collectionapparatus willbe washed with 10-20 ml deionized water and the fmal volume of urine willbe brought to 45 ml with additional deionized water. Daily fecesweight willbe recorded for each animal. At the designated times, aninws willbe euthanized by CO@ and gross necropsy performed. During necropsy, blood (m 6 ml) willbe collectedvia the abdominal aorta and transferredto blood collectiontubes without anficoagulant.Blood samples willbe allowed to clotfor a period of 15 to 30 minutes at room temperature, and the clotwillbe spun down in a centrifugeat 1100 x g for 5 minutes. The serum willbe transferredto labeled 1.5 rnlmicrofuge tubes and centrifugedagain at 2000 x g to removeanyrem&bg redbloodceus.Eachserasmplewillthen be transferretdo a separatelabeledpolypropylenemicrofugetube and flashfrozeninfiquidnitrogen.Liver,Iddneysand subcutaneousfatfrom each animal willbe removed, weighed, flashfrozen inliquidnitrogen and placed individuallyinto labeledsterilesample bags. The remainder of each carcass willbe placed ina labeledziplock bag and frozen (-70 'C) untilanalysis. Specimen Handling: Specimens willtemporarilybe stored in a freezersetto maintain -60 to 80"C. For metabolite analysis,these specimens willbe packed in dry ice and shipped to: 4 KrisHansen, Ph.D. 3M EnvirormentalTechnologyand SafetyServices 935 Bush Avenue St.Paul,NN 55133-3331 Ph:612-778-6081,FAX: 612-778-6176. 01/07/99 T-7098.1 POSF PK The 3M EnvironmentalLaboratorywillmanage theextractioannd analysis of sera,liveru,rineand fecesfortheparentcompound, POSF, and its presumed metaboliteP,FOS. AU tissuesampleswillbe retainedforpossible futureanalysiosf totalorganicfluorin(eTOF) ifdeemed necessaryby the StudyDirector.3M EnvironmentalLaboratoryor itsdesigneewould performthisanalysisA.U resultwsillbe providedforinclusioinnthefinal report. The number,type and dateofcollectioonf specimensto be generatedfor analysisareas follows: Table 3 - Specimens Specimen day I post dose Serum 10 Liver 10 Kidneys 10 Subcutaneousfat 10 Carcass 10 Urine 6 I Feces 16 day 2 post dose 6 16 day.4 post dose 10 10 10 10 10 6 6 day 15 post dose 6 6 day 29 post dose 1 10 10 10 10 10 16 Total 30 30 30 30 30 30 30 Data Analysis: Data collectedon tissuelevelsofparentcompound and identifiabmletaboliteswillbe analyzedforto3dcoldnetipcarametersand forstatisticaslilgynificadnitfferencebsetween groups usingStudentsT-testand/orANOVA. Responsibilides: Deanna Nabbefeldand Andrew Seacatwillbe responsiblfeordosingtheanimals, collectinign-lifsepecimens,performingthenecropsyand collectinagnd sendingtissue samplesforanalysis. o KrisHansen,3M EnvirommentaLwillbe responsiblfeoranalyticalnalysis. Andrew Seacatwilldrafta finarleportand ensurethereportreceivesappropriat3eM reviewbeforea finalreportisissued. 5 Signatures. Dr.@ddrew Seacat SeniorResearch To3dcologist Study Director L7 Deanna NabbefeII4MS Advanced To3dcologist .StudyToidcologist Sponsor Representative ate Date Date 01/07/99 T-7098.1 POSFPK 6