Document 5bYKjxB6G23qQ3K2z0qG8Bgy8

Argus PK Outline 5/03/2000 One Generation -R-eproductioSntudy of PFOS in Itats Pharmacoldnetic Analy Study Outline Study Objectives:This studywillserveasa follow-upstudyto Combined Oral(gavagg) EtrlW. Developmentaland Perinatal/Postna-tRa-lQroductioTnoxicityStudyof PFOS i Rats(ArgusProtocol418-008,3M studynumber T-6295.9).The objectivoef thestudy isto gainadditionaplharmacokinetc(PK) dataon PFOS inpregnantratsand theirpups. GLP Status:A GLP studywithappropriateQA audits;a signedQUA statementtobe includedinthefmal report. Aninuds: Female Harlan CD rats. Dose Groups: Six:VehicleControl;0.1 mg/kg PFOS; 0.4mglkg PFOS; 0.8mg/kg PFOS; and 1.6mg/kg PFOS. Number Per Group: 36 pergroup = 150 total Test Article:Compound willbe furnishedby theSponsorwho isresponsiblfeor compound identitaynd purity.Reservesample tobe retained. Compound Adndnistration:Females only,dailyoralintubationd,osevolume 5ml/kg. The vehicle(0.5% Tween 80 indeionizedwater)win be preparedweekly.PFOS WM be prepareddailyas a suspensionindeionizedwater with0.5 % Tween 80.Compound administratiownillstar6t weeks priortomating and continuethroughday 21 of lactation. Samples ofdosingpreparationwsin be collecteodnce duringthesixweeks priorto mating,onceduringgestatioannd onceduringthepostnataplerioclA portionof eachof thesesampleswillbe frozenand shippedtotheSponsorforpossibleanalysis.A backup sample of eachwillbe retainedby thetestinfgacilityR.ecordswillbe maintainedon how dosingpreparationwsere made. ClinicalObservations:Twice daily-approximatelyone hour afterdosingand again4-6 hours later. Body Weights: Weekly exceptwhen femalesarecohabitingwithmales duringmating. Food Consumption: Weekly exceptwhen femalesarecohabitingwithmales during mating. Mating: After6 weeks of compound administrationo,ne male (Argus breedercolony) willbe co-inhabitewditheachfemale.A femalewillbe determinedasmated upon the presenceof sperm positivevaginalsmear. The day ofmatingshallbe consideredday 0 of gestation. Page I of4 D.L Luebker Argus PK Outline 5/03/2000 Day Zero of GestationSacrificeand Examination: On day zeroof gestation6, pregnantfemalesineachgroupwillbe sacrificebdy CO, asphyxiationA. terminalbody weightforeachdam willbe obtainedand recorded.Animals willbe necropsiedand any grosslesionswillbe describedand recorded.Serum and liversampleswillbe takenfrom each dain.Allspecimenswillbe frozen(-20*C)and shippedon dryiceto KrisJ. Hansen, Ph.D.,3M EnvironmentalTechnology and SafetyServices9,35 Bush Avenue, Building2-3E-09,SL PauL MN 55133-3331.Upon receipts,erawillbe analyzedfor PFOS and liverswillbe storedforpossiblefutureanalysis. Day 15 of GestationSacrificeand Examination: On day 15 of gestation6,pregnant femalesin eachgroup willbe sacrificebdy CO. asphyxiatioannd cesarean-sectiwoinllbe performed.A terminalbody weightforeachdam and a pooledterminalbody weightfor each littewrillbe obtainedand recorded.Dams willbe necropsiedand any grosslesions willbe describedand recorded.Serum, liverand milk sampleswillbe takenfrom each dam. Dam sera,liverm,ilkand whole pups willbe frozen(-20*C)and shippedon dry ice to KrisJ.Hansen,Ph.D.,3M EnvironmentalTechnology and SafetyServices9,35 Bush Avenue, Building2-3E-09,SL Paul,MN 55133-3331.Upon receipts,era,milk and whole pups willbe analyzedforPFOS and liverswillbe storedforpossiblefutureanalysis. Pre-BirthSacrificeand Examination: Justpriortodelivery(morningof day 21 of gestation6)pregnantfemalesin eachgroup willbe sacrificebdy CO, asphyxiatioannd cesarean-sectiownillbe performed.A terminalbody weightforeach dam and a pooled terminalbody weightforeachlittewrillbe obtainedand recorded.Animals willbe necropsiedand any grosslesionswillbe describedand recorded.Serum and liversamples willbe takenfrom eachdam and herpups (pupsamplespooledby litterM)i.lk samples willbe collectefdrom each dam. Allspecimenswillbe frozen(-20*C)and shippedon dry icetoKrisJ.Hansen,Ph.D.,3M EnvironmentalTechnology and SafetyServices9,35 Bush Avenue, Building2-3E-09,SL Paul,MN 55133-3331.Upon receipts,eraand milk willbe analyzedforPFOS and liverswillbe storedforpossiblefutureanalysisM.AY ADD ADDMONAL DAMS IN SOME DOSE GROUPS SO WHOLE PUP PFOS LEVELS CAN BE EYAMINED OR MAY SAVE REMAINDER OF EACH PUP AFRER LIVER AND SERA ARE COLLECTED. Female ParturitionR:emaining ratswillbe allowedtodelivertheirlitters. Pup Exandnations:Pups willbe counted,sexed,weighed and observedfor moribundity/mortaliotny postnatadlays 0, 1,2,3 and 4,7, 14 and 21. Rats Found Dead or Moribund: Ratsthattheor aresacrificebdecauseof moribund conditionor abortionwillbe examined forcause ofdeathormoribund conditionon the day theobservationismade. Abortedfetusesand/orin-uteropups willbe examined tothe extentpossibleW.hen possiblet,hefollowingspecimenswillbe colloctoadnd analysis performed: Rats willbe necropsiedand any grosslesionswillbe describedand recorded.Serum and liversampleswillbe taken(individuallfyoreach dam and herpups - pup samplespooled by litterM)i.lk sampleswillbe collectefdrom eachdain.AR specimenswillbe frozen(- Page 2 of 4 D.J.Luebker Argas PK Outline 5/03/2000 2ooC) and shippedon dryicetoKrisJ.Hansen,Ph.D.,3M EnvironmentalTechnology and SafetyServices9,35 Bush Avenue, Building2-3E-09,SL Paul,MN 55133-3331. Upon receipts,eraand milk willbe analyzedforPFOS and liverswillbe storedfor possiblefutureanalysis. LactationDay 7 Sacrificeand ExandnatiOn: Sixdams ineach dose group and allof theirpups willbe sacrificebdy Co, asphyxiatioonn lactatiodnay 7. A terminalbody weightforeach dam and a pooledterminalbody weightforeachlittewrillbe obtained and recorded.Animals willbe necropsiedand any grosslesionswillbe describedand recorded.Sermn and liversampleswillbe takenfrom eachdam and herpups (pup samplespooledby sexforeachEtter)M.ilk sampleswillbe collectefdrom each darn. Allspecimenswillbe frozen (-20'C)and shippedon dry iceto KrisJ.Hansen,Ph.D.,3M EnvironmentalTechnology and SafetyServices9,35 Bush Avenue, Building2-3E-09,St. Paul,MN 55133-3331.Upon receipts,eraand milk willbe analyzedforPFOS and livers willbe storedforpossiblefutureanalysis. LactationDay 14 Sacrificeand Exandnation: Sixdams ineachdose group and allof theirpups willbe sacrificebdy CO,,asphyxiatioonn lactatiodnay 14. A terminalbody weightforeachdam and a pooledterminalbody weightforeach littewrillbe obtained and recorded.Animals willbe necropsiedand any grosslesionswillbe describedand recorded.Senun and liversampleswillbe takenfrom eachdam and herpups (pup samplespooledby sexforeachlitterM)i.lk sampleswillbe collectefdrom each dam. Allspecimenswillbe frozen(-20'C)and shippedon dry icetoKrisJ.Hansen,Ph.D.,3M EnvironmentalTechnology and SafetyServices9,35 Bush Avenue, Building2-3E-09,St Paul,MN 55133-3331.Upon receipts,eraand milkwillbe analyzedforPFOS and livers willbe storedforpossiblefutureanalysis. LactationDay 21 Sacrificaend Examination: Sixdams ineachdose group and allof theirpups willbe sacrificebdy CO, asphyxiatioonn lactatiodnay 2 1. A terminalbody weightforeach dam and a pooledterminalbody weightforeach littewrillbe obtained and recorded.Annnals willbe necropsiedand any grosslesionswillbe describedand recorded.Serum and liversampleswillbe takenfrom eachdam and herpups (pup samplespooledby sexforeachlitterM)i.lk sampleswillbe collectefdrom each dam. Allspecimenswillbe frozen(-20PC)and shippedon dryiceto KrisJ.Hansen,Ph.D.,3M EnviroranentaTlechnology and SafetyServices9,35 Bush Avenue, Building2-3E-09,St. Paul,NN 55133-3331.Upon receipts,eraand milk willbe analyzedforPFOS and livers willbe storedforpossiblefutureanalysis. Page 3 of4 D.J.Luebker rA oz @-3rA %o tz CL qQ tz 0 r co > PO ZJ'4 KV -(47-%o4,,U-) @7, twz CD F- Fr t:f 00 CrQ 0 > U.) W W En t2o too U ow. CD I m- Co 0 rA c7N CD tA 0 0 PO co U3 c7A L,) Zi !:@3 qQ U) 0 ts o C7\ 0 t-i Zi -3 Cr L, qQ 0 - -4 U) 0 w4 4 0 co) 'net- o co@ all0 w (71 0\ 0% 0% (7,0