Document wr9LdJRwR1o3OaJ73p7LZqLpV

! -3 Evian|Technology pPeO Box 33331 yr Study Title Oral (Gavage) Pharmacokinetic Study of PFOS in Rats PROTOCOL Author Lisa Clemen Date: June 8, 1999 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 JRE Laboratory Project Identification FACT-TOX-110 U2849 2 3M Environmental Laboratory 0977 Page 10f 10 Protocol #FACT-TOX-110 Study Identification Oral (Gavage) Pharmacokinetic Study of PFOS in Rats Test Material Perfluorooctane sulfonic acid potassium salt (1-6295) sponsor 3M Toxicology Services - Medical Department 3M Center, Building 220-2E-02 St. Paul, MN 55144-1000 Sponsor Representative Marvin T. Case, D.V.M., PhD. 3M Toxicology Services Telephone: 651-733-5180 Facsimile: 651-733-1773 Study Director Kristen J. Hansen, Ph.D. 3M Environmental Technology : and Safety Services Building 2-3E-09 651-778-6018 Study ILnocvaitvoioTne(sst)ing Facility Analytical Testing Laboratory Rg : ig Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA. 19044 3M Environmental Laboratory `Building 2-36-09 935 Bush Avenue St. Paul, MN 55106 . 60978 3M Environmental Laboratory Page 201 10 Sub-Contract Laboratory ProposSetuddSytIundityiaTtiiomneDtaatbele Study Completion Date Protocol #FACT-TOX-110 `Advanced Bioanalytical Services, Inc. 15 Catherwood Road Ithaca, NY 14850 Battelle Memorial Institute 505 King Avenue Columbus, Ohio 43201-2693 June 8, 1999 `August 8, 2000 1. Stupy Oral (gavage) pharmacokinetic studyof potassium perfluorooctane sulfonic acid (PFOS) in rats. 2. PurPOSE "This analytical study is (PFOS) in specimensof desig liver ned to determine levels and serumofrats. The of pota in-life ssium portio perflucrooctanesulfonate nofthis study was condiicte d at Argus Research Laboratories,study #418-013. All serum sampleswillbe extracted and analyzed at Advanced Bioanalytical Services, Inc. and all liver samples extracted and analyzed at `Battelle Memorial Institute. Additional analyses may be performed at the 3M Environmental Laboratory as methods are developed and validated. Ifadditional analyses are performed an amendment to this protocol will be written. 3. REGULATORY COMPLIANCE This study will be conducted in accordance withthe United States FoodandDrug. Atdhamtinainsatlryasitsioofn,thGeootedstLmaabtoerraitaolrmyiPxrtaucrteicfeosrSctoanncdeanrtdrsa,tiFoinn,alsoRluublieli2t1y,ChFomRog5e8,newiittyh,tahneedxcsteapbitliiotyn will not be conducted, and is the responsibilityofthe Sponsor. - R35 g <2 gE3 4. QUALITY ASSURANCE The 3M Environmental Laboratory Quality Assurance Unit will review the protocol and audit `study conduct, data, and final report to determine compliance with Good Laboratory Practice Standards and with 3M Environmental Laboratory Standard Operating Procedures. The QA Unit at the sub-contract laboratory will audit their study conduct, data, and results report priorto submittitnog the 3M Environmental Laboratory. 0097"9 3M Environmental Laboratory Page 30f 10 Protocol #FACT-TOX-110 5. TEST MATERIAL 5.1 Refer to Argus Research Laboratory protocol for study #418-013. 6. CONTROL MATRICES 6.1 Identification Rat liver and serum and/or rabbit liver and serum, traceability numbers will be recorded in the raw data and included in the final report 6.2 Source Argus Research and/or Sigma Chemical 6.3 Physical Description Rat liverand serum and/or rabbit liver and serum 6.4 Purity and Stability Not applicable 6.5 Storage Conditions Frozen at -20C+10Cor -50C10C 6.6 Reserve Matrix for as long as the A portionofthe control quality of the preparation matrix will be retained affords evaluation, but in the 3M not longer archives than ten years following the effective dateofthe final test rule (if applicable). 6.7 Disposition Matrices will be retained at the 3M Environmental Laboratory per GLP regulation. Certain matrices (feces, urine, and blood) may be disposed after QAU verification. 6.8 Safety Precautions Refer to MSDS for chemicals laboratory attire, and follow adequate precautions for used. `Wear appropriate handling biological materials and preparing samples for analysis. 7. s|oF EXE 2 psSs REFERENCE MATERIAL 7.1 Identification Potassium perfluorooctanesulfonate (PFOS), lot #s 171, 215, or 217 (equivalent lots) 7.2 Source 3M Specialty Chemicals 7.3 Physical Description White powder 7.4 Purity and Stability Puity of PFOS is 99% or greater. Stability has not been. determined. 7.5 Storage Conditions Room temperature 7.6 bReesreetraivneedMaastelroingalasAthreesqeuravlietsyoafmtphleefprroempaeraactihobnaatfcfhoordfsPeFvOalSuautsioend,ibnutthinsotstluodnywgeirll than ten years following the effective dateofthefinaltest rule (if applicable). 7.7 Disposition Unused reference material will be retained for use by the 3M Environmental Laboratory and will be discarded when the qualityofpreparation no longer affords evaluation. 00980 3M Environmental Laboratory Pago 40f 10 Protocol #FACT-TOX-110 7.8 SlaabfoerattyorPyraetctiarue,tianodnsfolRleofweradteoqMuaStDeSprfeocraucthieomniscaflorshuasneddl.inWgebairolaopgpircoaplrimaatteerials and preparing samples for analysis. 8. TesTSysTEm RRaetsseawrecrhepursoetodcaoslt#h4e18te-s0t1s3y.stTehmeafnedmwaleereramtasiwnitlalibneedgiavnedndtohseetdeastsmdaetsecrriailbeodricnontthreoAlrognucse daily beginning 42 days prior to cohabitation and continue through day 14 or 20ofpresumed. gestation. See table 1 for more dosage information. Table 1 Dosage Levels, Concentration, and Volumes Dosage Group Number of female rats Dosage mg/kg/day Concentration | Dosage volume mg/kg/day ml/kg ES TS Ewe [S J E for ee ls B Joa e Tess Is] fr ne Gwe qe [32 tem fee fs] Js | 9. SPECIMEN AND SAMPLE RECEIPT Tshpeec3imMenEsnvoifrtohnemfeonltlaolwiLnagbobroadtyortyiswsiulels raencdeifvleuihdosmforgoemnetihetyinsdaimcpalteeds pfooirndtossien atnhaelsytsuidsya.nSdee table 2 for specimen information. All specimens will be packed on dry ice for shipping. Table 2 Specimen Information Body tissue/fluid Collected Expected # of specimens "Seru--m Dam and Fetus animals |Dam-Predose, Days 7, 15, and 21 | 320 Dam and Fetus-A terminationofthe study _| 320 Fetus (pooled) Urine and feces -- Dam animals | Predose, Days 7, 15, and 21 320 urine and 320 feces EN Tiver -- Dam and Fetus animals | Dam and Fefus-At termination of | 80 Dam and the study 80 Fetus (pooled) 2158 [Milk Secreting Glands --Dam "At the termination of the study FINE" | animals 3 [Amniotic Fluid -Dam animals _|Day Sanabwy2i gig so | 00951 3M Environmental Laboratory Pago 50/10 Protocol #FACT-TOX-110 "Table 2 Cont. `Specimen Information Body tissue/fluid Collected Expected # of `specimens Embryo's with Placentae ~ Dam Day 15 and Day21 animals [CaPrlaccaenstsa--sF--etFuetsuasnainmiamlal | DAtayth1e5tearnmdinDaatyio2n1ofthe study Tiver/lung -- Fetus animal "At the termination of the study Total numberofexpected specimens: 2000 `Total numberof test animals: 64 `Total numberofcontrol animals: 16 I 160| [80 | Specimens sent to 3M Environmental Laboratories will be received and tracked according to applicable Standard Operating Procedures. 10. PREPARATORYMETHODS 10.1 FACT-M-1.1, Extraction of Potassium Perfluorooctanesulfonate or Other Anionic Fluorochemical Surfactant from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry . Electrospray/Mass Spectrometry 10.2 ETS-8-4.1, Extractionof Potassium Perfluorooctanesulfonaotre Other Fluorochemical Compounds from Serum or Other Fluid for Analysis Using HPLC- 10.3 If preparatory methods other than those listed above are used, anamendment to this protocol will be written. Any deviations from these methods will be documented and included with the study data. 10.4 Ifanalyses are sub-contracted to other laboratories, an amendment will be written to include their methods and copiesof each method will be attached to this protocol. g El 2 g] 'S $f g 11. ANALYTICAL METHODS 11.1 FACT-M-2.1, Analysisof Fluorochemicals in Liver Extracts Using HPLC- `Electrospray/Mass Spectrometry 11.2 ETS-8-5.1, Analysis ofPotassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum or Other Fluid Extracts Using HPLC-Electrospray/Mass Spectrometry 11.3 Ifanalytical methodsother than those listed above are used, an amendment to this. `protocol will be written. Any deviations from these methods will be documented and included with the study data. 11.4 Ifanalyses are sub-contracted to other laboratories, an amendment will be written to include their methods and copiesof each method will be attached to this protocol. 00982 3M Environmental Laboratory Page sof 10 Protocol #FACT-TOX-110, 12. DATA QUALITY OBJECTIVES `The number of spikes/duplicates, use of indicators are includedinthe analytical mseurtrhoogdast.esI,naandddiitnifoonr,mtahteiofnololnowoitnhgercdriatteariqauwailliltybe met: 12.1 Linearity 20.98 12.2 Limitsof detection / quantitation a) Sem: 175 ppb 12.2.1 Method Detection Limit (MDL) for PFOS. b) Liver: 15 ppb 12.2.2 tLhiemictaloifbrQautainotnictuartvieo:n (LOQ) ~ Equal to the lowest acceptable standard in 12.3 Duplicate acceptable precision <30% for the method 12.4 Spike acceptable recoveries 70% - 130% 12.5 cUosnefiromfactoornyfmiertmhaotdo.ryIfmaectohnfoidrsmatIonrdyetmeertmhinoadteissuasmepdl,easnwailmlebaedmree-natnatloytzheids using a protocol will be written. 12.6 Demonstrationofspecificity Chromatographic retention time, mass spectral daughter ion characterization. 13. SUB-CONTRACTED ANALYSIS 13.1 All analyses as detailed in this protocolwill be performed at 3M Environmental Laboratories, Building 2-3E-09, 935 Bush Avenue, St. Paul, MN 55106, at Advanced Bioanalytical Services, Inc., 15 Catherwood Road, Ithaca, NY 14850, or at Battelle `Memorial Institute, 505 King Avenue, Columbus, Ohio 43201-2693. 13.2 An amendment to this protocol will be writtenifanalyses are performed at laboratories other than 3M Environmental Laboratories, Advanced Bioanalytical Services, Inc., or Battelle Memorial Institute. 14. STATISTICAL ANALYSIS Averages and standard deviations will be calculated.`Thestatistical methods that will be used are. described below: 5[28 141 Data transformations and analysis Data will be reported as the concentration 5 (weight/weight or weight/vol) of PFOS or metabolite per tissue or fluid. Fi = aM Environmental Laboratory C020983 Page 70110 Protocol HFACT-TOX-110 14.2 Statistical analysis Statistics used may include regression analysis of cwointcheinnteraatcihodnossoevgerroutpi.me,Iafnnedcesstsanadrayr,dsdiemvpilaetisotnatsisctailccaulltaetstesd, fsourcthheascoSntcuednetnratt'itontesst, may be applied to evaluate statistical difference. 15.RepoRT A report containing all the resultsof the study will be prepared by the 3M Environmental rLeapboorrtataonrdy.suIbfmaintaliysteostahree3sMub-EcnovnitrroancmteendttaoloLtahberorlaatboorraytofroiresi,ncclaucshiolnabinortahteor3yMwiElnlvpirreopnamreenatal Laboratory report. Each report will include, butnotbe limited to, the following, when applicable: 15.1 Name and addressofthe facility performing the study 15.2 Dates upon which the study was initiated and completed 15.3 A statement of compliance by the Study Director addressing any exceptions to Good Laboratory Practice Standards 15.4 Objectives and procedures as stated in the approved protocol, including any changes in the original protocol 15.5 The test substance identification by name, chemical abstracts number or code number, strength, purity, the Sponsor and composition or other appropriate characteristics, if provided by 15.6 Stability and the solubilityofthe test substances under the conditions of administration, ifprovided by the Sponsor 15.7 A descriptionofthe methods used to conduct the test(s) 15.8 A descriptionofthe test system 15.9 A of description the data ofany circumstances that may have affectedthe qualiotry the integrity 15.10 The nameofthe Study Director supervisory personnel involved and the namesofother in the study scientists, professionals, and 15.11 A descriptionofthe transformations, calculations, or operations performed on the data, a summary and analysisofthe analytical chemistry data, and a statement of the conclusions drawn from the analyses go 15:12 Statistical methods used to evaluate the dats,ifspplicable 23S 15.13 The signed and dated reportsofeachofthe individual scientists of other professionals ERE involved in the study, ifplicable 15.14 The location where raw data and the final report are to be stored g 00334 `3M Environmental Laboratory Page sof 10 Protocol #FACT-TOX-110 15.15 A statement prepared by the Quality Assurance Unit listing the dates that study DinisrpeeccttoiroansndanMdanauadgietsmewnetre made, and the datesofany findings reported to theStudy Ifit is necessary to make corrections or additions toareport after it has been accepted, the changes will be made in the formofan amendment issued by the Study Director. The amendment will clearly identify the parotf the report that is being amended, the reasons for the `amendment, and will be signed by the Study Director. 16.LOCATIONOFRawDATA, RECORDS, AND FINAL REPORT Original data, or copies thereof, will be available at the 3M Environmental Laboratory to facilitate audits of the study during its progress and before acceptanceofthe final report. When the final report is completed, all original paper data, including those items listed below, will be retained in the archives of 3M Environmental Laboratory for at least a periodoftime asspecified by regulation, and as established by 3M Environmental Laboratory StandardOperating Procedures. 16.1 The following raw data and records will be retained in the study folder in the studylproject archives according to 3M Environmental Laboratory Standard Operating Procedures: 16.1.1 Approved protocol and amendments . 16.1.2 Study correspondence 16.1.3 Shipping records 16.1.4 Raw data 16.1.5 Approved final report (original signed copy) 16.1.6 Electronic copies ofdata 16.2 The following supporting records will be retained separately from the study folder in the archives according to 3M Environmental Laboratory Standard Operating Procedures: 16.2.1 Training records 16.2.2 Calibration records 16.2.3 Instrument maintenance logs 5 16.2.4 Standard Operating Procedures, Equipment Procedures, and Methods a 00985 3M Environmental Laboratory Page 9of 10 Protocol #FACT-TOX-110. 17. SPECIMEN `Specimens will be RETENTION maintained in the 3M Environmental Laboratory specimen archives for a `period of ime as specified by regulation or as long as the quality ofthe preparation affords evaluation, but not longer than ten years following the effective dateofthe final test rule (if applicable), and as established by 3M Environmental Laboratory Standard Operating Procedures. 18.PROTOCOL AMENDMENTS AND DEVIATIONS `Planned changes to the protocol will be in the form of written amendments signed by the Study pDriorteocctoolr aanndd wtihlelSbpeonastotra'csheRdeptoretsheentfaitnailvep.roAtomceoln.dmAellntcshawniglelsbteoctohnesipdreotroecdoalswpialrltboeftihndeicated in the final report. Any other changes will be in the formofwritten deviations, signed by the Study Director and filed with the raw data. 19. ATTACHMENTS 19.1 AttachmenAt Preparatory and analytical methods 20. SIGNATURES Marvin T. Case, D.V.M.,, Ph.D, Sponsor Representative 7 Date 4 for i = L/r8/39 Kristen J. Handen, Ph.D, 3M Environmental Laboratory Study Director _ Date gs 5g 3M Environmental Laboratory 00986 Page 1001 10 Oral (Gavage) PharmaSctokuidnyetTiictlSetudy ofPFOS in Rats PROTOCOL AMENDMENT NO. 1 AmAeugnudsmte1n2,t1D9a9t9e: 3M EnvironPmeerntfaolrTmeicnhgnoLlaobgoyr&atSoarfyety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification ET&SS FACT-TOX110 LIRN U2849 = ggg = 3M Environmental Laboratory 0987 ProtocAomleFnAdCmTe-nTtOXNo1.101 This amendment modifies the following portion(s) of the protocol: 1. PROTOCOL READS: Section 2.0 states this study is designed to determine potassium perfluorooctane sulfonic acid (PFOS) in specimensofliver and serumofrats. AMEND TO READ: This study is designed to determine PFOS in specimensofrat liver, serum, and urine. All Day -1 and Gestation Day 21 rat urine specimens will be extracted and analyzed by the 3M Environmental Laboratory. REASON: The urine extraction and analytical methods were not validated and approved prior to protocol approval. 2. PROTOCOL READS: Section 6.0 listsrator rabbit liver and serum. AMEND TO READ: Rat or rabbit urine from 3M Toxicology with a physical descriptionofrat or rabbit urine. REASON: The rat urine matrix was added af theptrotorcol was approved. _|s hZK 2 42'SS "B'S 2 3. PROTOCOL READS: Section 10.0 and 11.0 list the following methods to use for extraction and analysis: FACT-M-1.1 "Extractionof Potassium Perfluorooctanesulfonate or Other Anionic Fluorochemical Surfactant from Liver for Analysis Using HPLC-Electrospray/Mass FSpAeCcTt-rMom-e2t.r1y""Analysisof Fluorochemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry" AMEND TO READ: The extraction and analytical methods to follow at the 3M Environmental Laboratory are: ETS-8-6.0 "ExtractionofPotassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry" ETS-8-96.0 "Extractionof Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Urine for Analysis Using HPLC-Electrospray/Mass Spectrometry/Mass ESpTeSc-t8r-o7m.e0tr"yA"nalysisof Potassium Perfluorooctanesulfonate or Other Fluorochemnical ECToSm-p8o-u9n7.d0s i"nAnLailvyesriEsxotfraPcotstaUsssiinugmHPPeLrCfl-uEolreocotcrtoansepsrualyf/oMnaatses oSrpeOctthreormFelturoyr"ochemical Compounds in Urine Extracts Using HPLC-Electrospray/Mass Spectrometry/Mass Spectrometry" 3M Environmental Laboratory 00958 Protoc`oAlmFeAnCdTme-nTtOXNo1.101 REASON: The extraction and analytical methods FACT-M-1.1 and FACT-M-2.1, respectively, were updated on 07/22/99 to ETS-8-6.0 and ETS-8-7.0. Methods ETS-8-96.0 and ETS-8-97.0 were not validated and approved until after the protocol `was approved. 4. PROTOCOL READS: Section 10.4 and 11.4 state thatifthe analyses are sub-contractteod `other laboratories an amendment will be written to include these methods. AMEND TO READ: The extraction and analytical methods to follow at Advanced Bioanalytical Services will beattachedto the protocol. `The extraction and analytical method to follow at Battelle Memorial Institute is: "Method for Analysisof Perfluorooctane Sulfonate (PFOS) in RatSera by LC/MS/MS, Version "1p 0 O Liver "REASON: The analytical methods at the sub-contract laboratories were not included in the original protocol. 5. PROTOCOL READS: Section 12.2.1 b) Liver method detection limit is 15 ppb. AMENDTO READ; 12.2.1 b) Liver method detection limit is 8.50 ppb (ng/g). 12.2.1 ) Urine method detection limit is 1.5 ppb (ng/g). REASON: The validation supporting methods ETS-8-6.0 and ETS-8-7.0 included a lower `method detection limit for PFOS in liver. A validation inurinewas performed,afterprotocol `approval, to support this method detection limit. O@ue sly ig kn 125g #= aM Environmental Laboratory 0989 Amendment Approval qd Now Tour Marvin Case, D.V.M., Ph.D., Sponsor Representative Protoc"oAlmFeAnCdTm-enTtOXNo1.101 [1 Auge 1727 Date bi g Kris J. Hansen, Ph.D., Study Director )i8)99 Date sofg hs s1%3 3MEnvironmental Laboratory 00990 Study Title Oral (Gavage) Pharmacokinetic Study of PFOSinRats PROTOCOL AMENDMENT NO. 2 Amendment Date: September 28, 1999 Performing Laboratory 3M Environmental Technology& Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 BsbR 8 Laboratory Project Identification ET&SS FACT-TOX110 LIRN U2849 Fhe 3M Environmental Laboratory 00991 Protoc"oAlmFeAnCdTm-enTtOXNo1.102 This amendment modifies the following portion(s) of the protocol: 1. PROTOCOL READS: Section 2 states that all liver samples will be extracted and analyzed at Battelle Memorial Institute. AMEND TO READ: `All dam liver specimens (female mother) and 27 fetus (pooled) liver specimens will be analyzed at Battelle Memorial Institute. REASON: The fetal liver/lung is noat target matrixandwill not beextractedor analyzed. 2. PROTOCOL READS: : Section 9 states that 80 dam liver specimens (female mother) and 80 fetus (pooled) liver specimens will be sent to the Environmental Laboratory. AMEND TO READ: Seventy-three dam liver specimens (female mother), 27 fetus (pooled) liver specimens, and 81 fetal liver/lung specimens (27 liver/lung specimens sampled in triplicate) were sent to the `Environmental Laboratory. REASON: `The numberofanimals changed during the courseofthe study. 3. PROTOCOL READS: : Section 12.2.1 a) serum method detection limit is 1.75 ppb b) Liver method detection limit is 15 ppb. AMEND TO READ: The method detection limits for all compounds and matrices will be taken from the methods used for extraction and analysis. IE Bas) "REASON: "The method detection limits listed are specific to the 3M Environmental Laboratory. Statement wasaddedto allow for sub-contracted analyses and/or revised methods. i g aM Environmental Laboratory 00992 Protoc"oAlmFeAnCdTm-enTtOXNo1.102 4. PSeRcOtiToOnC1O6LsRtaEtAesDSth:at the original data,o copies thereof, wil be available at the 3M Environmental Laboratory to facilitate auditsofthe study during its progress and before acceptanceofthe final report. When the final report is completed, all original paper data, diantcal,udaipnpgr:oavpepdrofivneadl rperpootrotc,oellaecntdroanmiecncdompeinetsosf,dsattuad,yrcaoirnriensgpornedceorndcse,,csahliipbpriatnigornerceocrodrsd,sr,aw imnestthroudmsenwtilmlabientreentaaninceedliongsthaenadrcshtiavnedsarodfotpheer3atMinEgnpvriorcoendmureenst,aleqLuaiboprmaetnotryp.rocedures, and AMEND Section TO 16 READ: states that the original data, or copies thereof, will be available at the 3M EacncveiprtoannmceenotfalthLeabfoinraaltroerpyortto. fWachielintatteheaufdiintaslorfetphoert sits ucdoympdluertiendg,itasllproroiggriensasl apnadpebrefdoartea, idantcal,udaipnpgr:oavpepdrfoivneadl rperpootrotc,oalnadnedlaecmternodnimcenctopsi,essotfuddyactoarwrielslpobnedreentcaei,nesdhiipnptihnegarrecchoirvdess,orfas the 3M Environmental Laboratory. All comesponding training records, calibration records, `imnestthroudmsenwtimlaibnetreentaaninceedliongst,hestaarncdhairvdesoopfetrhateinfgacpirloitcyedpuerrefso,remiqnugipcmaecnhtapnarloycseidsu.res, and RCleaarisfoicna:tionofthe disposition of archived records ifanalyses are performed ata sub-contract laboratory. 5. PSeRcOtiToOnC1O7LsRtaEtAesDSth:at specimens will be maintained in the 3M Environmental Laboratory specimen archives. ASpMeEcNimDeTnsO wRiElAlDb:e maintained in the 3M Environmental Laboratory specimen archives. All Lsapbeocriamteonrsysuepatonocsoumbp-lceotnitornaoctfalnaabolryastiosriaensdwislulbbmeisrseitounnoeftdhtoetshueb-3cMontErnavcitrolanbmoernattaorly(s) ofriniaglinraelpocrhta.inThofecsupsetcoidmyeansndwirlelcobredsreotfursmteodrawgietchotnhdeitfioolnlsowwihnigledoactutmheenstuabt-icoonn:tratchte fsaicginlietdy. RCleaarisfoinca:tionofthe dispositionofdocumentation when shipping specimens for analyses 5 of performed by a sub-contract laboratory. Fe 3M Environmental Laboratory 00992 Amendment Approval Protoc"oAlmFeAnCdTm-enTtOXNo1.102 Marvin Case, D.V.M., Ph.D., Sponsor Representative fit Me Kristen J. Hansen, Ph.D., Study Director Date So 1997 . Date Bsh gs #i aM Environmental Laboratory 60991 " Study Title Analytical Laboratory Report on the DeterminationofPerfluorooctanesulfonate (PFOS) Presence and Concentrationin Serum, Liver,andUrine from the Gavage Study of T-6295.12 PROTOCOL AMENDMENT NO. 3 Amendment Date: 20 January 2000 3M EnviroUnrmiennetaAlnTaleycshensology and. SFalfueotryinSeeArnvailcyetsical Chemistry Team B9u3i5ldBiunsgh2A.v3e.n0u9e St.Paul, MN 55106 PerformLiivnergALnaalbyosresatories B5a0t5teKlilengMeAmvoerniuael Institute Columbus, OH 43201-2653 AdvancedSBeiroaunmalAyutisclaylseSservices, I1n5c,Catherwood Road Ithaca, NY 14850 sk ERE Ne s Laboratory Project Identification ET&SS LRN-U2849, ArguFsACStTudTy:OX-41181-0013 3M Medical Department Study: T-6295.12 g 3M Environmental Laboratory 00995 APmreontdocmoelntLRNNu-Um2b8e43r9 This amendment modifies the following portion(s) of the protocol: 1. PROTOCOL READS: The study director Hansen, Ph.D. for the present study was identified in the protocol as Kristen J. AMEND TO READ: TD.hVe.Mr.o,lePohf.sDt.udaysdoifre2c0toJrafnouratrhye 2p0r0e0s.enTthsetupdryevwiaosusrsetausdsyigdnireedcttoor,MaKrrviistnenT.J.Case, Hansen, REASON: has been reassigned to the role of Principle Analytical Investigator. TGhoeodroLlaeboofrsattuodryy dPirraeccttiocrewSatsanrdeaarsdssigtnheadt oiuntalinneefsfotrutdtyopeenrssuorneneclomrpeqluiiarnecmeewnittsh (refer to 21 CFR Part 58), 2. PROTOCOL READS: The sponsor for AMEND 70 READ: the present study was identified as Marvin T. Case, D.V.M., Ph.D. The role of sponsor for the present study was Ph.D., as of 20 January 2000. reassigned to John L. Butenhoff, REASON: Tspooennssourrreoltehawtatsheresatsusdiygndeidr.ectIonrtdhiosemsannonteral,sopecrasrornyntehlerdeustpioenssiobfilsittuiedsy sponsor, and the workload are more evenly balanced. =e she gi: 3M Environmental Laboratory 60996 Amendment Approval JPlroitoncoel pLAiNm-Us2a04d9 Flan X. Bali John L. Butenhoff, Ph.D., Sponsor Representative Febpuny 22000 Date bite for -- KristeJn. Hansen, Ph.D., Outgoing Study Director =Feb --2000 Date 7 0/2 : Marvin T. Case, D.V.M,, Ph.D., Incoming Study Director ate HBRET 3M Environmental Laboratory 000997 Study Title Oral (Gavage) Pharmacokinetic Study of PFOS in Rats PROTOCOL AMENDMENT NO. 4 Amendment Date: 20 April 2000 Performing Laboratories < 2g rs e528 IE Laboratory Project Identification ET&SS LRN-U2849 FACT TOX-110 Argus Study: 418-013 3M Medical Department Study: T-6295.12 600998 APmreontdocmoelntLRNNu-mU2b8e4r49 This amendment modifies the following portion(s) of the protocol: 1. `PTRheOTamOeCnOdLedREsAeDctSi:on 2.0 text states that this study is designed to determine PFOS in specimensofrat liver, serum, and urine. AMEND TO READ: This study is designed to determine PFOS in specimensofrat liver, serum, feces, and urine. REASON: oTfhteheansatluysdiysofoflfleocwailngtitshseueisfsouratnhceeotfartghetecphreomtioccoall.aFnedc/eosr eitxstarnaactliyotnesanwdasanaadldyteidcatlo mtehthsocdospe were not validated and approvedpriorto protocol approval. 2. PROTOCOL READS: `The amended section 6.0 lists rat or rabbit liver, serum, and urine. AMEND TO READ: Add: rat or rabbit feces with a physical descriptionof rato rabbit feces. REASON: Analysisoffecal tissue the study following the for the target chemical issuance of the original and/or its protocol. analytes was added to the scope of 3. PROTOCOL READS: Section 13.1 lists allofthe laboratories that will be conducting analyses for this study. AMEND TO READ: Add: Centre Analytical Laboratories, Inc., 3048 Research Drive, State College, PA 16801 REASON: Feces analyses were added to the scopeofthis study. The sub-contract laboratory performing analyses was not in the original protocol. 4. PROTOCOL READS: Sections 10.4 and 11.4 state thatifthe analyses are sub~contracted to other laboratories an `amendment will be writen to include these methods. AMEND TO READ: The feces extraction and analytical method used by Centre Analytical Laboratories will be; 00M-023-003 (Revision Feces by LC/MS/MS." 2), "Determinationof Fluorochemical Residues in Monkey/Rat Sp Ressow: & The sub-contract laboratory performing feces analyses was added to the scopeofthis study; # NE this method was not validated and approved prior to protocol approval. 5 EPs 3M Environmental Laboratory 20999 rPirotsocsoleLrANi-nuzo84o0 Amendment Approval Gln BFE John L. Butenhoff, Ph.D., Sponsor Representative Spe! (3, 2000 Date ae toe Marvin T. Case, D.V.M., Ph.D., Study Director femdom Date iS5 ofthe Sire ab Environmental Laboratory 01000