Document 4aoz6gXB80xqeN4Jp1mGwarrx

#601 --_ ~ @ ~ AR226- 3429 33 000272 | ARAR6 = 3429 D> EXPOSURE ASSESSMENT PROTOCOL Work RequestNo.: Service Code No.: Performing Institution: Study Title: 15076 1361 Haskell Laboratory for Health and Environmental Sciences OEcxtpaonsouirceAAcsisdeAsnsimoenntinotfhMeeMaasnuurfeadctSuerreuomf APemrmflounoiriunmated Perfluorooctanoate (APFO) at Fayetteville, NC | | |Ji 3), | 2) || | | | || Study Director: Coinvestigators: SJeonsieoprhIFn.duVsitsrkioaolilH,ygJir.enCiIsHt H1a0s9k0elEllkLtaobnorRaotaodry for Health and Environmental Sciences Newark, DE 191714 S Hh-- e-- ------ Robin C. Leonard, Ph.D. HPraisnkceilplalLaEbpoirdaetmoiroylofgoirsHtealth and Environmental Sciences EDlutPoonntRoCaodm,pPa.nOy. Box 50 Newark, DE 19714 Mary A. Kaiser, Ph.D. Research Fellow Corporate Center for Analytical Sciences -- HSa.sMkaelrlk LKaebnonreadtyor,yPfho.rDH.ealth and Environmental Sciences 1090 Elkton Road Newark, DE 191714 pD Company Saniized.Doosnotcontain TSCACB 0002773 \ )) qd || "o Study Sponsor: | | Prk Jennifer Locklear Fayetteville Works DuPont Company 22828 NC Highway 87 West Fayetteville, NC 28306 EE Bonnie G. Wilson, RN Fayetteville Site Nurse Fayetteville Works 2D2u8P2o8ntNCComHipgahnwyay 87 West ay -- NC 28306 ------ SDaHvEidMMa.naRguerrak DuPont Fluoroproducts ---- BMP CRP 711-2216-C `Company Sanitized. DossnotcontainTSCACB 2 000274 @) ' bmg i) } || 3 || || BD Por David. Fura `SHE Manager Fluoroproducts AUTHORIZATION FOR SERVICES FroJmoseph F. -- viso--ct, Gi -- `Senior Industrial Hygienist Haskell Laboratory Requested By: Date: December 12, 2008 Tiler Exposure Assessmenotf Measured Serum Perfluorinated Octanoic Acid Anion in the Manufacture of Ammonium Perfluorooctanoate (APFO) at Fayetteville, NC Authoritiys requested to undte her stut dyia denk tife ied above and described as follows: Ey ------ APFO in Fayetteville, NC. for assessment of the effectiveness of the industrial hygiene `controls measures in pltoa minc imieze exposureto APFO. Thesecondary purpoofsheiscczupatonal exposure rect tocharacerAiPzFeO `exposureinworkersengaged inthemanufactureofAPFOandtodetermine 1)whether the majorrouteof exposureisvia inhalation or dermal, 2) the lengthoftime required for the compountdo reach steady-state, and 3)ifthere isa sufficient population, determine the length of time for serum levels to return to baseline following cessation of exposure. `This will be accomplished by longitudinal sampling and analysis of serum PFOA anion levels according to a repeated samples design, air monitoring, and estimating dermal `and oral contributotoiveornalsl exposure. Correlations between external and internal measures of exposure will be calculated. The data will be examined for within- and between-subject variation associated with specific tasks on the process line. `AUTHORIZATION FOR SERVICES Additional Support Required from the Plant: Personnel ii -adtioal IH esos to concur af Samping an vcsiate Air sampling equipment including pumps, calibrators, battery eliminators, ..... process upsets, data anomalies IapNpIrTovIaAlT-IEOsNtiDmAatTeEd:apIpmrmoevdailatdealtyeupon|DEeScTeImMbeArT3E1D, 2C0O00MPLETION: December 15,2003 ToeDe200m s mo a | 5BONSE SS AS GG0L ESTIMATED COST: December 15, ee `GENERAL LEDGER/SUBACCOUNT: [S-------- 000275 |@ beIfltoiws,raenqudersettmurene.tswithyourapproval,enterthegeneral ledgerandsubaccount,sign AUTHORIZEDamon nedZP(G odwhaores2fis/o 3 . -& "ow 00027 . TabloefContents Page No. L REVIEW BN APPIOVAL cc... Principal Investigator and CO-NVESHGaIONS .........vrrrs + LB pmesmmmom---------- nT Definitions OEPINES usin] + BEKGI erin] UL SUVEUNANCE MEOUS...cocosssssns + PRCIPENSOBsrscmssmmmmemmosmssennn3 DoRCONBR msmmmimmmmmmmninmlommsdosammemrnd DR ANGIYSIS rrr, 11 | Ey EU | | 3 Factors That May Influence the SurveilanceDesign | * HUM@NSUDJECE PIOECHON....r.errrrssrmrssessmrmris1n2 1 Protocol Amendments and DEVANONS .........rmmcn 12 | + UE ASSEIRIOR ommmismmimminmommmmngagm 18 | V. ReqUIred RESOUICES ......vurvsssnnns 13 VI. ArchiOvfSitndgy DOCUMENS.......crerrrrs 13 Vile SHOGAPNY wsmmmsssssmmsissmmssnsiimmpnsdsmaoinn 18 ExygenBlood Method ClaAyirtMeothnod Wipe TestValidation BD) \ 4 000277; @) | | 1 | 9 | VII Appendix Appendix 1: ~ Standard Operating Procedure: Blood Collection + Appendix 2: Standard Biomonitoring Samples Operating Procedure: Process for Analysis of Blood + Appendix 3: Standard Operating Procedure: `Study-Specific Standard COopenrsaetnitngFoPrrmosc,edaunrde:CoDlolceucmteionnta&tiPorno,ceAscscoiunngtoafbiSlietryu,manSdamPprloecsedures for * Appendix 4: Standard Operating Procedure: Documentation,`Accountability, and Sample Chain of Custody Requirements for the `Shipping of Serum Samples * Appendix 5: Blood Collection Checklist Appendix 6: Air Sampling Strategy + Appendix Industrial H7:ygSiteannedaLradboOrpaetroartyinagndPrFoiceeldduWroer: kQuality Assurance Procedures: + Appendix 8: Standard Operating Procedure: Sample Handling, Chain-of- Custody , Appendix Preservation and 9: Analytical Meth Shi odfor pp--inAigr Samples Air Samples Appendix 10: Air Monitoring Data Form Appendix 11: Perfluorooctanoic Acid Wipe Test Analysis BD) 5 000275 &) REVIEW AND APPROVAL STUDY DIRECTOR AND COINVESTIGATORS | attest that,to the best of my knowledge, this protocol is scientifically sound, describes the HstyugdiyenaendPrpacrtoicceesd,utrhteeosrebseearucshedprwoittohcoalpparnodparinaytearmeegnuldamteonrytsretqhueirreeomfe.nts, Good Industrial `Submitted: LZ orSoyVBioor Git owe_sshshis Approved: -- YOupkino,FefHLoaLasbit D Date:_/-o/s I) | tf 6 000279 &) INTRODUCTION Abstract This occupational exposure study will characterize APFQexposurebymonitoring PFOA anion in workers engagedinthe manufacture of APFO and to determine 1) whether the major routeof exposure is via inhalation or dermal, 2) the length of time required for the. compound to reach steady-state, and 3)ifthere is a sufficient population, determine the length of time for serum levels to returntobaseline following cessationof exposure. This will be accomplished by sampling and analysis of serum PFOA anion levels according to a repeated samples design, air monitoring, and estimating dermal and oral contributions to overall exposure. Correlations between extemal and internal measures ofexposurewill be cwiatlhcuslpaetceidf.icThtaeskdsatoanwitlhlebpereocxeasmsinliendef.or within- and between-subject variation associated 1D) Definitions: i 'APFO --Ammonium Perfluorooctanoate | Biomonitoring -Industrial Hygiene serum samples analyzed for PFOA anion used as an indication of occupational exposure LC - Liquid Chromatography MS -- Mass Spectrometry PFOA ~ Perfluorooctanoic Acid PFOAAnion --perfiucrooctanoate anion, the analyteof both PFOA and APFO by LC/MS/MS Ap ) Objective: The objecotfitvhies study is to monitor serum levels of PFOA anion levels in manufacturing and manufacturing hygiene controls. support workers of APFO as a measure the effectiveness of industrial BACKGROUND Despite the absence of any persuasive scientific evidence establishing acausal association between exposure to PFOA and specific health effects, there continues to be 7 000280 | @) questions raised on the health affects of PFOA. Epidemiological studies on the health effects PFOA exposure have focused on occupational populations. A curently validated analytical method (with a limit of quantiication as low as 0.5 ppb) allows increased understanding of background levels. In Fayetteville APFO production began in the fall of 2003. Prior to start-up baseline blood serun samples were taken. With this background, we have a unique opportunity to monitor `exposure as afunction of processtime. This PFOA anion in serum levels will be measured routinely as an indication of the effectiveness of industrial hygiene controls. Itis our goal to minimize employee exposure to APF in the AFPO manufacturing `process and to assure compliance with the DuPont Acceptable Exposure Limit (AL)of 0.01 mgm'. D} || BD E 000281 8) SURVEILLANCE METHODS Pi ICIPANT SELECTION 1 i | lo D | { AD Biological monitoring evaluates the absorption of PFOA/APFO by measuring the chemical or its metabolites inbody fluids. Serum sampling is the preferred method for biological monitoring because there is avalidated analytical method for measuring PFOA anion, rather than a metaboliteorthefluoride fon in this medium. Workers in the APFO manufacturing process are chosen because they are likely to have highest exposuresatthe Fayetteville Works,wewereableto get a pre-start up baseline, and wecan track serum levels with time from the process start-up. DuPont's goalisto minimize exposure. Workers employed in the new APFO production unit at Fayetteville Works and for `whom a baceline level of serum PFOA anion was determined before beginning work in the unit will be included in this surveillance study. Participants will be all APFO workers at the DuPont Fayetteville Works sitewhovolunteer for participation in this study, and sign a consent form. DATA COLLECTION : Every employee assigned to the APFO manufacturing area will asked to participate in the [PFOA anion blood monitoring program. The employee will be asked to sign aconsent form `and a baseline blood sample will be taken before the employee begins work in the APFO manufacturing area.Afterthe baseline sample is collected, the employee will participate in the normal 6-month sampling program. Once in the blood monitoring program, the employee isexpectedto have blood drawn and analyzed every 6-monthsforthe durationofthe study -- `whether or not they remain in the APFO manufacturing area. For each participant, date of birth and date of hire will be collected. For each task with potential for exposure to APFO, job title, work area and begin and end dates will be recorded. 5 000282 | Atotal of 9 job titles and 5work areas have been identified as having potential for exposure o ") to APFOatthe Fayetteville Plant. The number of unique task/work area combinations with potential for APFO exposure vill be determined. D0 i | | i BH ) BloodSampling `Standard Operating Procedure: Collecting and Processing Blood Sample--s Appendix - 1 Standard Operating Appendix 2 Procedure: Process for Analysis of Blood Biomonitoring Samples `Standard Operating Procedure: Study-Specific Standard Operating Procedure: DProoccuemsesnitnagtioofnS,eArcucomuSntaambpilletys,~aAnpdpPernocdeid-xur3es for Consent Forms, and Collection & C`SutsatnoddayrdROepqeuriarteimnegnPtrsofcoerdtuhree;ShDiopcpuimnegnotfatSieornu,mAcScaomupntlaebsil--tyA,ppaennddSixam--p4le Chain of Blood Collection Checklist ~ Appendix -- AiSrampling Air Sampling Strategy - Appendix - 6 `LSatbaonrdaatrodryOpaenrdatFiinegldPW roceo d~urAepr :pQeunakdliixty A-s7surance Procedures: Industrial Hygiene `Standard Operating Procedure: Sample Handing, Chain-of-Custod,y Preservation and Shipping -- Ar Samples -- Appendix -- 8 Analytical Methodfor AirSam-pApplendeixs--9 Air Monitoring Data Form -- Appendix -- 10 Wipe Test Perfluorooctanoic Acid Wipe Test Analysis -- Appendix - 11 o 000283 . BD DATA ANALYSIS mee exposure data will be analyzed as follows: 1. The general statistical procedures described below will be used in the analysis of the. data. Analytical results of the regular samples will be analyzed using appropriate `statistical methods. 2. Descriptive statistics will be generated on each exposuremetricwith all measurements pooled. 3. Where there is sufficient data, regression of serum PFOA anionforeach individual will be done on gender, age, most frequent task , 4. Analysis of variance for each time point measuremenotf serum PFOA anion will determine the between- and within-individual variation, and 5. Where there is sufficient data, estimate blood clearance rates of PFOA anion in workers by continued surveillance following cessation of exposure | |@ ~D } | | EXPOSURE ASSESSMENT Exposure will be measured by the following parameters: 1. Serum level of PFOA anion, priotor beginning work in the APFO Manufacturing Area and measured every six months (per the unit schedule) 2. Personal air monitoring of each participant, selected to cover all tasks and all shifts with potential for exposure to APFO 3. Area air monitoringforthe laboratories, the manufacturing lines, and the packing and shipping of APFO 4. Estimate the dermal contributiontothe biomonitoring result for each worker, and 5. If necessary estimate the contributionof other routes of exposure. ) The investigators will compile a document for each area that will summarize data related to APFO exposure: (1) descriptions of processes where exposure to APFO may occur, including natureof and datesof engineering controls, changes in operating procedures and potential exposure points, (2) specific tasks performed in work areas where exposure to APFO may occur (3) work area names, job titles and time periods with potential for exposure, (4) documentation of personal protective equipment use, (5) air sampling data and analytical methods employed, (6) plant production records and (7) details on work conditions and practices gained from interviews and observations. u 000284 L 4B) Because dermal contact may be a significant route of exposure, estimatesofthe amount and frequencyof skin contact wil be used to estimate a dermal dose using the suriace area of the body covered, frequency of contact, length of contact and results of the surface wipe tests. Personal air monitoring data wil be used as a direct measure of inhalation exposure. FACTORS THAT MAY INFLUENCE THE SURVEILLANCE DESIGN AND ANALYTIC METHODS Analytic methods are new and recently validated. Only one laboratory is GLP certified to do the blood analysis and only one laboratory is currently AIHA accreditedforthe air sample analysis. Timing of results could become an issue. `The sizeofthe surveillance population will notbe large thus limiting the statistical power. | `uTsheedwaispeatqeusticaknaclhyesciks ifsorccoomnptlaemxi.naStaimopnloerttuhreneafrfoeuctnidvteinemseswiolfl anodtecaolnlotwamwiinpaetitoesnts to be | procedure. ~~); HUMAN SUBJECT PROTECTION Haskell Quality Directive QDO07-C-001 requires compliance with the U.S. Federal regulation known as the Health Insurance Probabiity and Accountability Act of2003 (HIPAA) and is supported by standard operating procedures that specily privacy and confidentiality requirements. PROTOCOL AMENDMENTS AND DEVIATIONS Intended (planned) changes to the protocol will be made by protocol amendment. Unintended or unexpected divergence from this protocol will be documented in the field raw data as a deviation. The Study Director will be notifiedofall protocol amendments and deviations as s00n as possible. All changes in of revisionsofthe approved protocol and the reasons for the changes will be formally preparedbythe Study Director, signed and dated by the Study Director and Sponsor, and maintained with the protocol. AD |" QUALITY CONTROL dFaietlad tsotaHfafswkielll lreLcaobrodraotbosreyrvwietdhienxspeosvuerneddaaytsaoofncosltlaecntdiaornd.iHzaesdkfeolrlmsstaaffnwdilslernedvitehwe foobrsmesrvaendd `rceocnotnaccitleedmiif snseicnegssorardyistcorreepsaonltviendfiosrcmraetpiaonntw,hmeirsseinagp,proorpriinactoen.siSsatmenptledadtaw.orDkaetraswwiillllbbee transmitted electronically along with selected hardcopy data to the Study Director. n 000285  | ~le~, |g REPORTING OF RESULTS S`mtoantiutsorrienpgorrtessuwlitlsl abnedreuvpioenweredcbeiyptthoefSbtluodoydDirreseuclttosr.fAolflionwailnsgumtmhearryeceriepptoorfttwihlel bqeuaritsesruleydaaitr htyhgeiceonnicstluwshioonwiolfltchoemsmtuundyi.caAtceotmhmeumnitcoatthieosntoufdythpearftiinaclipraenstusl.ts will be to the site industrial RESOURCES TO CONDUCT THE STUDY `Sampling Equipment `maintenance log ~ 10 Air Sampling Pumps, chargers, battery eliminator, calibrator, and PfieerldsoonbaselrAvsastiiosntsa,nceen--sAurneeaxdpheerrieenncceedfointdhuisstrpiraoltohcyoglieannisdtitnovecsotnidguactet daiervisaatmipolnisngf,ropmrotvhiede protocol. See Air Sampling Stra--tApepengdiyx 6 ARCHIVING OF STUDY DOCUMENTS "cTohmeppurtoetrocporli,ntaomuetsn,damnedntfisnatlortehpeorptrwotiolcoble,arracwhidvaetda,thcroonusgehntthfeorsmtsa,ndaanraldytpircoaclerdeusruletss,of the Haskell Laboratory Information Services Group. BIBLIOGRAPHY Reference Exygen Blood Method Reference Clayton Air Method Wipe Test Validation AD) n 00028 | D! || | | | D, ---- Appendix-1 1. BLOOD COLLECTION SCHEDULE: confi tPherdsoantaelstcoobmemugniincsaatmpiloinnsgw.iBlllobeodmasianmtpaliensediwlitbheedarcahwsntuedvyeprayr6t-imcoipnatnhts(,0To cwiolvlerbealclohlilfetcst,edbplroioodrs0amapnleesmpwliolybeee cwoolrlkeictnegditnhthienar:ea.10T-hdaeyepmepriloody.eBe awisllitnhesnajmopilnes the normal6month rotation. L1. Field Laboratory for Blood Collection Blood samples will be collected at Fayetevill Site Medical bythesite nurse. 12. Aborted Blood Collection: sIafmapnlianngicvpaaltbeed rs-asmcphlodiunilgsedcaansc-esloeodm,aw5oprokesrisbweilflcbre tnhoetiforiiegdipnarlolmypslcyhendudled bdalyo.odTfstahmeprleeaonne maesdcihceadluolpehdydsaiyc,atlhrnetahsaotnssttuhdayt csuobnjtecitn'dsiscaamptlhinegdwrialwlibneg of rescheduled 0occuronthe extdayha hesameorsimilartaskswil be performed. 2. PROCESSING BLOOD SAMPLES: SAetrthuemfseepdarItbioorna.tory,bloodsamples il nositambientformorethan 2horsprio 0 3. TRAVEL BLANKS: Fink "bTerasvheilpBpleadnwkistwhilcollbleecatneudnbulsooeddspaomlpylpersoptyoliennestuurbteshuaptphleiaemdpblyeEsxywgeerne.nTohticsobnatlaamiknwaitleld ~uring sipping. 4. SAMPLE IDENTIFICATION: "iTdehnetisiftieesnuhresfsiinteduasntdipaelrhsyogniesnaimsptlewidl.lTrheestcehains-aomfpcluisntgoiddyenftiofrimcawtiiolnrseycsotredmtthheat shipping date. 6. PROCESSING SAMPLES FOR FROZENSTORAGE AND SHIPMENT: 6.1. SAMPLE STORAGE: Appendi3x- Standard Operating Procedure: Documentation, Accountability, and Procedures for ConsentForms,and Collection &ProcessingofSerum Samples. 000287 | | ~ | | i ApD) 6AS.a2.pmppSleAiCMh:PaLiSnEaoSfaHCIraPMoEOsNpTRt:iangsPecnao:r DhoecuSmiepnitantgioofn,uAraccSoaumpnltes, nd 7. SAMPLE ANALYSIS: T-- hedete proceduresforalysis of studyspe orPFOAar presen 5. DATA ANAALNDYSTASTISITICSAL METHODS: r_at--o! otof throu slo will sly wing ppt sion 9. CONTROFL BIAS: iSesaiptitesnlaoserFreomsthfovrotlyaprrnilagtantoyfsiudafitns.leCntoomnpaosfoonsdof te demographics, workhistories, andusualtasksperformedofthe voluntaendetrhse abstentions will be made in order to identify any systematic differences in the two groups. 10. REQUIRED FIELD DATA RECORDS: iAnlkdlfbityneldsednaotepsearensmdoodnatnftea,esAtheyetaaastgmdueisgmtroubaendfibolelieodlnogukitd.perIobmm pryttley,papcn seccunrtatcedloayt,ra.dnAdiihnirdnede.slibdTtlhet FltsTwtill iin omg oars fl veri sos of so Following 1. rA doesvcriepticoinopfmtehnettawseksdperformed by the sampled participant, including personal 2. Arete aSiog ag na semge 5. Avopyoitn ssempehs r em go iversr ot. 4. PevnrcseeonsaaslnwioenrakgpobatsllipooorafoatxacalirwooeronkegxrodpivoiyngsiShSheomeusiiboetusachteatw:healnowasrhincg, 11. PROTOCOL AMENDMENTS AND DEVIATIONS: UniteIndteonrdeTad ( rEapgl Dceha earncgn eoswtin oiotvnhe nerporid ootgoi) cnolttorwuiaslcllbretamcaedoesbyerpmroeetnoctomlsanmaeehnddSmooenmtsw.etodnests v00288 L &) | ~B || PD) sCaohnoadnnmagaesispnwotisasliilnbbeledef.woAirtlmhlaltchlheyanppgreostriocnoeolrbpreyvatihsreiSoentsuoddfythDieraepcptroro,vseidgnpedanrddao atneddttbhyertoehaesScotnusdo fyoDritlrheector 12. REPORTING OF DATA: reports wUilplobn cpormepplaerteidonbyoftthheePrainnacliyptailcanlvaentidgfaiteodrsp,oUrpoonnscoofmtphleesttiuodny,oafntahleysteicraelpsonrtd,iacdlrdaft {fiinnall rreepporrtt wwiilllbbe psreedp.are for rview and approval byth Sud Director. Upon approval. a practices",Thweorrekpeorrtbawciklgrcoonutnadinanndatexipvereiednecsec,riapntdiobnseof pheertcianevnitrofnomremaatatlioconndciotliloencst,edwdourrking Each replicate (camps) ofthe sd, 1 addition, the inal epoct will clude: 1. Nianimtetaedndaandddcroemspsleotfehdefacilitiespecformingthe studyandthdataon which thestudywas 2. aObmjeencdtmiveenstsanidnptrhoecoerdguirneaslsptraotetdocolth approved protocol, inching any devon andor 3. Identifaindcdeasctriiptoionnof thetestandreference substance(s) 4. Adescriptionof llmethodsused 5. Arasnusmfomramraytioofn,thcealdcautlaagteinoenr,asteadswhsi,leocrolndeucrtoipnegrtathiesontsupdeyrafnodrmaeddeoscnritphteidoantoafall 6. "Thenameanddatedsigur oftheStyDirector 7. A descriptionofall conclsion drawn fom analysis ofthe samples 8. Complete descriptionofthtet system 9. Locationof aw da,Goareport, ndalspecimens 10. A verified copyofthe sud protocl including protocol deviations and amendments 11. Locoafttheitesotsinte Uponacceptofathnefcinealreport,all studyspecificoriginalrawdatawillbesubmitted tWoiltlhee tSphonpsroorpeanltoynogfwtihtheSepxoacntsocro.pSieusbosfenqoune-nsttsuudpyposrptecriefciocrsduspsphoarltldbaetac.opOireigdi,ncaerltdioficeudmaenstse:xact copies, and included with the original raw data and submitted to the Sponsor for archival. 13. QUALITY ASSURANCE: "Thefieldphase of hisprotocolwil beconductedanddocumentedascompletelyas possibletoensurethequalityandintegrityofthe datageneratedforthisportionofthe study. 000289 &) ELEorih,BaPre oltaylwGpbosrtonorfds tesotdyoowiloObspoifonTode incyoimppteivi he Gomdelt 14 RECORDS RETENTION: ReSporEt, d, cssin istyil ive adel onion 9) || . 000290 | BH ) L | ? Di || | 8. Appendix2 ProcessforAnalysisof Blood BiomonitoringSamples Preparation Phase: 1. Palllanotthseirtephraen-dslaempalpipnrgoparriraatnegceommemnutnsications, schedulingofphicbotomy, and 2. Plant site requesis supplies from Exygenatle4awseekts in advance; estimated number ofsamples is needed at this time. 3. Exygen amanges for supplies to be shipped and schedules samples or analysis 4. Supplies are shipped to plant site with instructions on processing and shipment. `Sampling Phase: 5. Blood is samanpdplroceessded into serum atthe plant ste: 6. Serum samples are shipped directly from the plant ite to Exygen laboratory `KweitnhnaesdaymopnlethleisstainmgpalnedliCshtainign.-of-Custody records. The plant will cc S. Mark AnA alyn sisaa ndlytRi eporc tinga Phasl e: 7. Exygen analyzes the samples and prepares a draft report to review. The draft reportandcopiesoftherawdataaresent toS. MarkKennedyforreview 8. dSr.aMfta,rakndKegnivneesdtyhercoentrvactthielaabenpalewyrtmiicssaslidoantat,omfiankaeliszeantyhecaonrarleycttiicoalnsr/euppogrrtadestto he 9. The contract lab finalizes the analytical report and sends ito S. Mark Kennedy. S. Mark Kennedy sends a copyofthe repart to Study Director Joseph F. Viskoeil who archives the originalreportat Haskell and informs the plant site industrial hygienistofthe results Completion Phase: 10. The ite handles appropriate communications at the site 11. Upon receipt of the invoice from the contract lab, S. Mark Kennedy will provide the appropriate (plant-site-specfic) charge code to have th bill paid againsta standing purchase order 12. Samples willberetained for 12 monthsperthe method validation. Samples willbediscarded afer 12 months upon the Study Director's approval. 000291 8) D} | | Appendix-3 Documentation, Acco`SupnetciafbiciSltiaatnnydd,arPdroOcpeedruarteisngfoPrrCoocnesdeunrte:Forms,and Collection `& ProcessingofSerum Samples I. SCOPE AND APPLICABILITY `This SOP governs the documentation, accountability, and procedures for consent forms, and collection & processing of serum samples. 2. REPONSIBILITIES 2.1 The Study Director is responsible for conducting the necessary training, covering proper documentation, accountability, and procedures for consent forms, and collection & processing of serum samples. 22The Phlebotomist who performs the blood collection is responsible. for ensuring that the Consent Form is completely and correctly filled out by each `participant prior 0 venipuncture. 2.3TfhoerPmshlaendbothteosmaims&ptleSsiatereInpdruospterrilaly cHoylglieecntiesdtaanrde lraebseploends.ible for ensuring that 3. Names Study Director: Joseph F. Viskocil, JrCIH Phlebotomist: Bonnie Wilson, RN- Site Nurse Fayetteville Site IndustrialHygienist; Jennifer Locklear 4. MATERIALS AND EQUIPMENT 4.1. ConsentForms (duplicate or triplicate) 42. Serum Separator tubes (Red top) ~(Supplied by Exygen) 43. Vacutainers, Needle holders and Needles--(Supplied by Exygen) 44. Centrifuges 45. Transfer Pipettes and Polypropylene Tubes--(Supplied by Exygen) 000292 ) _ 7 D. | i 1 ] 2D) 5. PROCEDURE 5.1. Hveanvieputnhcetupraer.ticipant complete the Consent form prior to initiation of 52. sEnasmuerceotdheatntuhemcboneasrentthfeobrlmoocdomspalmeptleedfbryomcatchhatppaarrttiicciippaanntt.receives the. 53. Original consent for each participant will be sent to Study Director for storage. 54. Accesstoconsent forms tobedecided 5.5. Collect blood specimen using usual venipuncture technique. Fill tube p------ 5.6. Gently invert the scrum separator tube 5timesto mix. 57. Allow bloodtoclot a minimumof 30 minutes but no longer than 1 hour. 58. Centrifuge for 15 minutesat2500to 3500rpm 59. Remove from centrifuge 5.10. Serum portionofthecollected sample in the SS tube is now ready to be. `pipetted into the specially provided polypropylene tubes, 5.12. Trheeadpyolfoyrprtorpanyslpeonret ttuobtehse(llaabbeled withspecimenID numbareenro)w 5.13. Pleasesec Appendix 2fordetailson howto completethe chainof custody form and prepare the sample for shipping. 5.14. Sampleswillneedtobefrozenimmediately at ~20Candstoredfrozen. 5.15. Samples willalsoneedtobe shipped frozen. See Appenxix-4. 6. REFERENCES - Appendix-4 7. FIGURES Figure 1. Consent form 000293  -- i || || || | PCORNOSGERNATMTO PARTICIPATE IN PFOA EXPOSURE ASSESSMENT Study tie: AEnxipoonsuinrethAessMeasnsumfeancttoufreMeoafsAumrmeodnSieurmumPePrefrlfluuoorroioncattaendoaOtceta(nAoPiFcOA)caitd Fayetteville, NC Ipnatrrtoidcuicptaitoino:n iYsovoulaunrteabrey.ing asked to participate in a research study. Your tYhoeusredrveicciessioynouwhreetcehievreothnrootuogh pDaurPtiocnitp'atseOwciclulphaatvieonnaol eHfefaelctthonPryoogurracmoamrpIlnotyemgernatteodr `HSepaelctiahliSsetrovricNeusr,soerBifetnheefrietsisAadnmiyntihsitnrgaytiooun.doPnlietasuendaesrkstqauneds.tions of your site's OH `WThheapturisptohseepoufrpthoesperoofgrthaims sitstuodlye?ar aboutth relationshipbetweenspecifictasks iadnednttihfeyewxapyossutroerleedvuecleotfhiPsFeOxpAosthuartemay25oclcouwrainlewvoelrkaesrpsaossitbhlies.plan ite, and to ATrheebtehneerfeiatnoyfbepnaretfiictispfatriongmipnatritsicsitupdaytiiisnntghtahtiysosutumdayy?gain important knowledge a"bThoiustkhnoowwleexdpgoesuwirlelshmealpyovuarryOHdeSppeecniadliiosntnsg, tihneduwstaryisalcherytgaiiennitsatssk,saanrdeceangriineederosutf.ind ways to reduce these polniial exposures. PRA WYhoaurtdpaoretsictihpiastisotnudiytihnivsosltvued?y may last up to 5 years. "PFOA. Nootherbloodlevewlilslbedetermined. W"Tihelmtoherespteuodpylbeeatfhfeacptaerdtiicfiypoatuedionnthoitspsatrutdiyc,tihpaetmeo?reaccurateand representattihve orteshuelrtssiwtilslifbew.eIhnaavdedietniooun,ghwepamrtaiycibpaentasblteo0pruosveidteheasgeoroedsusltasmtpolecosnitzreo.l exposures at Wh aretaherisk itsokfsinvolpartivcpatineg dwithibnetihnigs enroluilnltdebhiedstshteusdmya?ll risk of a (bruise) where the blooids drawn, Otherimportant temsyou shouldknow: Withdrawal fromthestudy:Youmay chootosteop your participation in 000294 |@ | dD | | efhfiescsttoundyyoaturaneymtpilmoey.meYnoturordtehceisseiornvitcoesstyoopuyoruercepiavretitchirpaotuigohnDwuilPlonhta'vse no Occupational Health Program or Integrated Health Services, or Benefits. Administration. New information: To the best of our ability, any significant new findings during this research study will be made knotowyonu. + Confidentiality: Every effort will be taken to protect the names ofthe participants in this study. However, there is no guarantee thattheinformation cannot be obtained by legal process or court order. Funding: Haskell Laboratory has been provided funding to conductthis study by the DuPont Fluoroproducts business. Number of participants: We expect Approximately 30 participants. Who shouldyou callwithquestions aboutthisstudy? Questions about this study may be directed to the site OH Specialist, Jennifer Locklear; oVirsktahoecisli,teaNtuHrassek,elBlonLnaiboerWaitlorsyo:n,WRN.SYouNmayduarlisongconnotracmtalthbeuSsitnuedsysDhioruercst,or, Joe `Wilyou be paid to participate in this study? No. Blood samples will be drawn during the work shift. CONSENT T`ohpapvoreturneiatdy tthoe aasbkoqvueesitnifoonrs.mat1iaognraebeotuot p(anrtaimceipoaftestiundtyh)isasntduhdayvaenbdeIenhagvievebneann given acoop fthyis consent document for my own records. Prnthame: Pass Number: Yousigawre Dues Witness: Date: _ | B) CompanySanitized.Doesnotcontaln TSCACBE 000295 | Appendix-4 &) TITLE PAGE `Standard Operating Procedure: Documentation, Accountability, and Sample ChainofCustody Requirements fS orthhe ipop fSeriumSnampg les Analytical Facility: + 30E5x8ygReenseRaersceharDrcihve Sue Cole BALcans D Analytical Facility Management: John Flaherty, Vice Presideat, Operations `Company Sanitized. Donote contsainTSCACB ||i Page 1 of7 000296  1 | || a) | | | I SCOPEANDAPPLICABILITY `rTehqiusiSreOmPengts foor ttvhheesdheoicpummmeennttonaftsiesornu,macscaomunptlaebsiltiotyt,heaanndalsyatmipclalecfhacailiintyo.fcustody 2. REPONSIBILITIES 2.1 TenhseuripnhgictbhoatotmhiestChwaihnoofpCeursfotromdsy AtnhaelybsliosodReqcoulelsetctFioonrmis(CrOeCsp)on(sFiibgluere for 1) `ispcaomcplakentaedlsgyhieapntdhecsoerrreucmtlysamfipllleesd.out and signed bystaffmembers who 222 Tthhaet tPhreincchiapailnoIfnvceussttigoadtyorraetmatihnesanianltyatcitcaalfefractihlietysaixurpelsepsoanrseibrleecfeoirveednsbuyritnhge analytical facility. 3. DEFINITIONS Analytical Facility Management; JEoxhyngeFnlaRheersteya,rcVhic3e05P8reRseisdeenatrcohfDOrpievrea,tiSotnast,e ry Principal Investigator:EExmyigleynDeRceskeearr,cShc,ie3n0ti5s8t Research Drive, State College, PA 16501 | | 8 Company Sanitized. DossnotconTtSaCAiCBnI Page 20f7 000297 @ | -~> 4 | | 2D 4. MATERIALS AND EQUIPMENT 4.1 ChainofCustody/Analysis Request Forms, triplicate design. 42. Aprorteeacdtiilvye eaqvauiilpambelnetsaopuprrcoeporfiadtrefyorhicaenpdellleitdsnr.gy Also, ice. necessary personal 43 Coolers project. numbered and They must be coded with a unique purchased from Uline identification assigned for this Shipping Supplies Specialists. `TheUlinepart corresponds to naupmabcekragfeorwthhiecrheqiunicrleuddecsooolneres is S-7888. 19" x 12" (This part number x 12.5" cooler and `one shipping box). 44 wPlialstiaclsboagbsefoorbiuzsieneads sfercoomndaUrliynse,amppalret pnauckmabgeirngSp-r1i7o0r7to3s"hipxpin5g%., These 4 mil reclosable poly bags, 10perc0arto0n). 5. PROCEDURE 5.1Bveerfiofrieed.colWlhecetnincgaacnhysbalmopolde, itshecoclolemcptleedtaedunainqduesiaglnpehdacnounmseernitc fiodernmtimfiucsattiboen c"oSdaempwliellIbdeentaisfsiicganteidont"occoalcuhmsnamopflet.he TChOisCnsuhmeebte.r wLiilklewbiesee,ntetrheeddaitnteoatnhde tthiamteotfhecascehrubmlosoadmspalmeplperecpoalrlaetcitoinon will be recorded, and is completed will be the date entered and time into the appropriate columnonthe COCsheet. 5.2.`Enaucmhbers,amwphliechcowoilllerbewiwlrlitbteenaosnsitghneecdooaleru.nique alphanumeric identification 5.3Biendfiovrieduaplalcykaignitnogs,matlhle plparsitmiacrbyagsserausmsescaomnpdlaery containers packaging. must The be placed secondary" packaging must ink. havetheunique sample code writtenon it in permanent black 5.4Bwielflorveertihfeystahamtptlheessaarmeppleasckdeodciumnednrtyeidcoen,tahpeeCrsOoCnofotrhmermtahtacnhtehxeacptalcykwaigtehr th"oCosoelbeerCionngtpeantcskeVedriifnitehd"epcrooolmeprt.. Theverifier theninitialsanddatesthe 5.5.aSwaampylaessartetoombienidmiiszpeertshedeipnosacsioboilletryfoulfloifmdpraycitc,eapenldlestus,chartrhaantgetdhienyasurceh w`ciotmhpdlreytielcyepceolvleertesd. wNiotmhotrhee ptehlalents2.0 sTahmeplceosolaerre tmousbetplbaecceodmpinleotneelycofoillelre.d mTihneimciozoelleorswsiolfldbrey iccoevteoresdubplrimoamtpitoln.y after packing with dry ice pellets to Page3 of7 000295 &) |> | | | | | | || | |B ) | 5.6.sThhiepmteinmte mbuesttwbeeenabthleteocobmepalcectoiuonnteodffosreorunmthpereCpaOrCatipoagne.andThpearcekamguisntg nfootr be any unexplained gap in the chainofcustody for the samples. 5.7.`Acfolmeprlettheed.seSrecutmiosna2mp(l"ePserasroennpelacIknavgoeldv,edthine SChoioplmeerntIDo#f Sparmopmlpets")wiollf tbhee C`TOheCprfionrtmedanlasomwei,lslibgneatcuormep,lientiteidalastatnhdidattiemoe,fisfiitghnaastnnouetrebdeeteonbpererveicooursdleyd. for each person who entered information anywhere on the form. The page number prompts also needtobe filled in. 5.8.The person who packaged the samples then completes the fist line in the "Relinquished by" section, and verifies that any unused prompts are lined out. 5.9f.aTxhenCuOmbCefroartmthiestthoepnfofatxheedftoorEm.xygAfetneRrietsiesafracxheSda,mtphelpeiRnekcceoipviynigsurseimngovtehde and becomes part of the clients records. The white original and yellow copiesaretobeplacedinanenvelope, whichisthenplacedinto the shipping cartonwiththecooler that contains thesamplesto whichthe COCform corresponds. 5.10R.ecTehiveisnegruatm tshaempaldedsraersesregqiuviernedattotbheeshtiopppeodfttoheExCyOgeCn Rfeosremarbcyh SFaemdpElxe s`covceerpnaigbhltedelivery. This is the only carrier and method of shipment that is 5.11. Upon procedure receipt will of the follow samples Exygen bSyOEPxyVg4e0nl,Res"cCaorncthr,actthoRcehsaeianrcohf cSuasmipoldey seHcatnidolni,nga".copAyfotferthEexCygOeCnwiSlalmbpelfeaxReedcebiavcikngtoctohmepclleiteenst.the "Received by" 5.12`.ExyAgneny wiplrlobbleemdsocuwmietnhtedde.liveTrhyeoErxycognednitPiroinncoipfalseImnpvlesetsiguatpoornwialrlrinvoaltifayt the clientofanyproblems immediately. 5.13. Correction and Clarification ofRawData 5.13.1. Absseoocnoarsamawndeyaitsnaf.orAmtatthiiosnphoainstb,etehnisefnotrermediosnnotthteobCeOdCifsocramrd,eidt tfroarnsacnryiberdeaosnotno.anoItfheirt sbheeecto,mbeusttdhaemtagread,nisnfcorrmparitoicpoenstsmwiailyolbbnee shdeoceutmmeunsttebdevaitatfaocothnteotode tohnettrhaensnceriwbepdapgea.ge.The original damaged 5.13.2.Iferrors are made in handwritten entries, orifclarifications are to `lbienem,a5d0ea,5 tnhoet etroroanbtsciunrfeotrhmeatoiroingiwnialllebnieryc.rosTsheedn,oudtepweintdhinagsuinpgolne Page4of 7 000299 PD 3 Ftihgeunraet2u)r,e oanfdtthheeeerrrroorr,caondeaipspirnoiptriiaalteedearnrdordcatoedde.iIsfcthhoeseernr(orscies. dmeotraielecdoemxpplleaxnatthiaonnwihlaltbiespcroovveirdeeddbbyytfhoeotneortmeorocnotdheleiCstO,Ca page. 6. REFERENCES Exygen Research SOP V401,"ContractResearchSampleHandling" 7. FIGURES Figure 1. ChainofCustody/Analysis Request Form Figure 2. LiofsErrtor Codes 8 SIGNATURE PD i an 7 Date Analytical Facility Management 8 Page Sof 7 000300 ) |1 | Lo { || | i i { | I) Figur1e. Chain ofCustody/ARneaqulesytsFoirms Exygewnh, CSHHAAINNOEOF CCUUSTSOTDYODY RCOLLENCTIONSSITaINFORMATION |a STAFF VOLVEE D I SAMPLE SHPIVENT TT ||=me ese |-- Paux LT=e= Tr __ ------ ---- tm 7 r r r r ] r rr r _-- . tr tt rt r rr y i [A Te SCE (= re 2 `CompanySanitzed.DoesnotcontainTscacar Page6of7 000301 &} | L| D | | `Figure2. ListofErrorCodes EXPLANATION OF ERROR CODING aCnodrrleocrtwirointeo-orvfae, adnadalisimnagdienbtyhedcroarrweicndagatsai.ngClearneemtursoubgehtathkeeeorrtoneeoeusndasta the the original entry. correction, and The correction is initialed and dated by an explanation is given for the correction. the individualmaking `The following error codes shall be used to explain corrections: (6) Late Bnry (Wl) Inadvertently recorded in Wrong Location (cc) Changed for greaterClarity (wo) Write Over (Technician inadvertently wrote over the errorinstead (sp) oSfpedlrlaiwnginegrraorline through the error) (ir) Inadvertently not Recorded at time.ofinitial observation ((c%e)) RCealccourldaitnigonEeErrorror ((ne)) NEuamteryrincoatlintirtainaslcerdiapntidodnaEtrerdorat ime ofcntcy ((twed)) WTrraonsncgriDpattieonenEtrrryor (om). StrayMarks appearing on data with no other feasible explanation (1) Technical error odCneottdaehisle,edCoatOthCtehrfeotthriammn.eotEfhxcomosorerrdesecfttihinaoetnd.caabmonvoe.tmbaeyebxpelauisneedd, uisfienxgplaanineerdroorr dceofdienmedusbtybfeooftunlotley: Errorcodesareto becircled ftehaesbiboltet,otmhoefterhreorpsahgeo.uld be andwrittenaboveorclosetothecorrection. Ifthisis not footnoted `with an asterisk`orother symbol, and explained at BD) Pawagee? 7of7 000302 L) Example COC Sheet andUse ofError Codes Exygeenn:, CHAIN OF CCUUSSTTOODDYY cai r Ear Es E REATE EIE EE EX e ey gts SeeERs EE ee EE cir > Ac|2._ZJohLe RDoociStEPP nr ets ne ---- 3.3. Smith _ Poov es.eue m y WY Tigres Amol Touhy wysm| shih dime fm | vb - | { | 9 har [A Tollszasem] ellis a'atm ER lui --aSaen ols Zap] = Tr "1 = |o -- mme m ctm-- m------ ------ ------p ------i-- 2 e] rred r= ------ aS-- Te FF [wa------ rer} = 1 SACH WANT SSCA S,W, A | eee ------ tf ieee} remem He D eeetmteeeeeee | re-- m--a e SPETETe Orrine S--a e --Po-- [EERE CONN SS as,Toe s, _IA_______ OFN orn] SS CSTE | 000303 L& | ~e)$ | | | | 3 Arps BLOODCOLLECTING CHECKLIST Eesane Epoesibtie Daten Tme Blood sampler Coletng Bind Samp FCie lemTsoefmoe e1r5 msiStss2400---35--0 7--0 v=e a!wo:rrl ~ a Sop(miliary time), A PipetSerumintoSPpeocilmeyn pNtrubmoebwepirtyhspleceimnenIeD . sTGPiehrmmeapmiscCtehodaonnaetyCyorwoendsyeCepirOnOedForupm21 YoeeCo(sOC3ror3thne0weNiyvope_ess_ > ae Serum Sales mkbesipped ren Ses vn YES NO Shiodeuscor"inis SDHiigo3s0e5p8RpeevDove State College, PA 16801 -- PisScoatwmomnii A(pipeoNndse)8)Name pri) ei ASchciopupnetdaFbeildtEyx,oavnedrnSiagmhptlOeNCLhYa.in(opefrCSutsatnoddayrdReOqpueirraetimnegntPsfroocretdhuereS:hiDpopciunmgenotfaSteiornu,m owe Fs Sedthrg BodCling Cress 0 Soph Vio3. CT NPOeBvoax,0DTE11R09701l4o05nc0Rorws `Company Sanitized.DoesnotcontainTSCACBI 000304  3) | | | ~D | | | || H) |\ Appendix 6 AIR SAMPLING STRATEGY Talhl eexgpooasluorfeesx.pWoosrukreersasmsaeyssbmeenetxpiossteodrewchoiglneipzeeraflolremxipnogstuarsekss,abnyditnocimdiennitmalize `bcyonottahcetrsf.roAmltbhaocukgghrtohuendhicgohnetsatmienxaptoisounr,eosrubseuaclaluysreesoufltnferaormbyatawsokrksepre'rsfoowrmned task, exposures significant can occur from theother two sources. `Serum levels of PFOA anion are an indication of overall exposure. Air sampling results are an indication of airborne exposure. aTihtebsoimgeniefxipcoanscueroefiesxcpoontsruorlelebd.ySiinncicdeentthaelncounmtbaecrtocfanjnobotasbseigansmseenstssedatuntil the aFasyseitgtnevmielnltesisesxmtaelnld, winetoweilvlesnaimnpgsleanedacwheejkobenadsss,igancmceonmtmqoudaratteirloyn.s Wwihllebreemjoabde to monitor workers at these times. tTewcohnAicPiFanOmoFniietlodr/sCtChReTCeocnhtnriocliaRnosoamreaonnd dtuhteyfootrheeratcehchn1i2c-ihaonurdosheisft.thOene fmiaejldowroirkto.yfSAinPcFeOt/hCeCfRieltdewcohrnkictieacnhnsiacmipanleiss mionrtehelifkieellyd.to have an exposure, the ProposedAirSamplingPlanforthefirst12-Months / C(oTlWleAc)t sqiuxar(t6e)rpleyrfsoornaelacsham1p2l-ehsouTrismhei-ftWescihgehdtuelde-(AvAePrFaOge Field/CCR technician) 24 samples C(oTllWeAct)sqiuxa(r6t)erpleyrsfoonra8l-hsoaumrpslheisftTismcehedWueliegh(tAePdF-OALvaerbage and Mechanical Technician) 12 samples C(olTlWeAct)tqhuraeret(e3r)lypefrosro8n-ahlosuarmsphlifetssTcihmeed-ulWeeifgohrtAePdF-AOverage Supervisors 3 samples C(olTlWeAct)tqhuraeret(e3r)lypefrosro8n-ahlosuarmsphlifetssTcihmeed-ulWeeifgorhtAePd-FAOveFrLaSge 3 samples Cfoolrletcecthsniixca(l6l)ppreorfsesosniaolnsalamppelresson(ndeulracthieonmoifsttaasnkd)eqnugairnteeerlrys) 6 samples 000305 I ) "Total 48regularly scheduled personal samples per quarter Sampling Strategy to be re-assessed after 2 quarters r`eSparmepsleinntgewdill be scheduled to ensure that offshifts and weekends will be proportionally WForAan hour shift; a minimum of 6- hours sampling is required to qualify as a 8-hour WForAa 12-hour shift; a minimum of 10- hours sampling is required to qualify as a 12-hour | Db | | ||| ~D 000306 ) APPENDIX 7 | | ~~ | | | | D1 QUALITY ASSURANCE PROCEDURES IN ) ID FIELD WORK Twhoirsk.procedure specifies quality assurance procedures for industrial hygiene field Laboratory Standard Operating Procedures Ttuobimnigniamnidzethtehaeirpsosasmipblilitnygofpucomnptsamsihnoautlidonb,e wexitpeemdadloswunrfwaictehsaof50ad/a5p0tseorlsu,tion of diisospproospeydl oaflcporhoople.rlTyh.eMwiinpiemsuhmoupledrsboenaalspsruomteecdtitvoebeequciopnmteanmtinsahtoeudldanidnclude safety glasses with side shields and nitrile gloves. "This proceduremustbe followed to assure equipment calibration and maintenance is done properly. Relevant quality control procedures are incorporated into this procedure. `Sample Numbering System Ewiallchasfsiiegldnsaanmuplmebemrusfotrbeeacahsssiugrnveeyd.aTuhneiqhuygeinenuimsbtewri.lTahdedianduunstirqiuael hsyegqiueennitsital number for each sample taken on a survey. Example: 82-24-01 82 Year of Survey 24 Survey number 01 Sequential sample number Field Sampling sStaremaanplslpeto.hretOCtaoVlaiSnbdrcaatfsesreostmatmeupssleoersipotute.hmepCrsasliwimibitrlhaatrceosusanamtmpeplrilsn,egopdruermvopitcsoemsbeetifenorprsleaasattnildceaabsfategbrsefefooarrce.h and after each survey. Before and after flow rates shouldbewithin 10%. cInosmtprluicatntceest. sVuobijdecatnsynostatmoplteasmwpheircwhitahrepesrussopneaclt.samplers and observe for Place sampler inlet in downward orientation to provent contamination Immediately after a sample is taken, place caps into fitter cassettes. 000307 @) cPruesasuare asnadmptleempveorlautmuerse 1corcraelcotuiaotnse tpop2b5coCncaenndtr7a6t0iomnsmoHfggamsuesstabnedmvaapdoerst,o for comparison with exposure limits. vPorlesusmuerseaarnedutseemdpteoractaulcruelactoerrmecgti/omns oafrpearntoitcumlaadtees, fbourtcaocmtpuaarliasironsawmitphle exposure limits. Control, Known and Duplicate Samples Cloanbtorraotlo,rky,noalwonnganwditdhuapilricsaatmeplseasm.ples mustbe submittedtothe analytical i`Cmomnetdriolatsealmypclleossaerdeacnodllseecatlioend,daevnidceassswihgincehd haasveequbeenteinalopneunmebedra.t Tthheeysitpsr,ovide a check on the analytical zero, qKunaontwintysoafmaplcehsemairceaclolaltecHtaisokneldlevLiacbeosr,awtohriych(ohraavtethbeeesintespuinkdeedrwtihteh haygkineonwisnts D dpirroevcitdieona),chcaercrikedononantahleytsiucravleayc,ouarnadcya.ssIingnseodmae sceaqsueesn,tiitalmanyumnboetrb.eTfheaesyible to - tphreepSatruedyreDliiraebclteokr.nown samples and they may be omitted upon concurrenocef aDsuspliigcnaetde sseaqmupelnteisalarneuamrbeearsa.irTshaemyplperosvtiadkeean cfhreocmksioxntso atmwpellivnegiannchdeasnaalpyatritcaalnd precision. cRheesmuilsstoifn vcoinetwroolf, tkhneoswpnecaifnidc daunpallyitciactaelsmaemtphloedsumsuesdttboedeevtaelrumaitneedawdietqhutahcey. dGuepnleircaaltley,sakmnpolwenvsaalmupelseshroesuulldtsbesh2o5ul%d obrelweistshaipna2rt5.%oRfetahseonsspifkoerddeavmioatuinotnsand should be determined and corrective actions taken. D| > 000308 I@ ) | | LD | | | (Ccowonhltilrcoohtlieovrneersduioslvtislcesoshsaotsutltrdhinebgeaenvtle)e.rsasgSteahsmaunprl5vee%yroesfsautlmhtpselseehxovpuoollsduumrbeee ofcifomrintroedncotsseoedlfooonrstchtoen.etrol sample actions traeskuelnt.s. Reasons for deviations should be determined and corrective bMeinsiubmmuimttreedq,uiarreedgniuvmenbeirn sthoeffcoolnltorwoil,ngktnaobwlnaa;nd du~plic~ ate samples which must NumberofSamples Field Control Known Duplicate Pairs <10 1120 2140 >41 1 3 3 4 > 2 a 4 0 1 2 2 Two Two at at oeancehleovfeltw*o* levels *** OAdndeitpiaoinralatpaeiaschmoafytwbeo lneeveeldsed**i*f*a wide range of concentrations is expected. INDUSTRIAL HYGIENE SURVEY RECORDS - Quantitative Survey Data AFiorrSmamwphliichngmuDasttab-eAselnatitrosatmhpelSintguddyatDairmeuctsotrbfoerepnetremraendeonntArierteMnotniiotno,ring Data feOoqtruhcieaprlmcQeuulnaatnttidioitnamste,inmvseuisoSntusrb,voeeryeontDtheaerterad-doaOnttahtewhrehidAacithraM,wiolsnluibtceohruaisnsgevdeDnaitntilaaatFqiouoranm,n.iriotaoimveanmdanner Survey Field Notes(otherthan quantitative data) diTnihfseocsrameradtneiodotnewshwieallrnebnetoasklueomnnmgoaenrrliynzeaeesddaeidnn.tihnHefoosrwumeravlvekerye,yrifetpoionrtmt.ehmeTohirenydsu.estfrSiiiaegldnihnfyoigtciaeensntimstasy. be. sjhuoduglmdenbte, aotrtigaicnhaeldnoton-AqiuraMnointiattoirviengnoDtaetsanFeoerdmtofobreperretmaainneedn,tttheenitnifoonr.mation 000309  Analysis and Analytical Reporting Phase: Crlepaoyrttoannadnacloypzieess tohfetshearmapwledsaatandarperespearnettso aMadrrayftA.reKpaoirstetro, rPehv.iDe.w.fTorheredvriaefwt. 1. cMoarrryecAt.ioKaniss/eurp,grPahdeDsrteovtiheewsdtrahfet,aananldytgiicvaelsdtahtea,comnatkraecstalnabypermission to finalize the analytical report 2. The coniract Kaiser, PhD. lab finalizes the analytical report and sends it to Mary A. 3. MF.arViyskAo.ciKlaiwshero, aPrhchDivseesndthseaorciogipnyaolfrtehpoerrteaptorHtastkoeSltluadnydDiirnefcotromr,s tJhoeseph plant industrial hygienist of the results - 000310  D | }E1 N | Appendix -3 ASIARMSPALMEPHLAENSDLING, CHAIN-OF-CUSTODY, PRESERVATION and SHIPPING -- s"Thhoeulpderlsiomnitciolslehcticnogntthaectsawmiptlhetshheosualmdplweesa.r nite disposableLa orexam gloves and `ichnSiaclmllpelda,etnhu,obwdeersyv(elOroVcsSaat)mopnwl.iellStbabemeppsrloaevnitdudbceeodsllbaeyncttdheecdoslclaoemnctptrleaedcstsesadhmopaulbleodsranatoraterboyeatsnrudebsqhcuotiureleddd bttooebesored temperature extremes. Ifnololrodweirngtopmroitnoicmoilzsehtohueldpobsesifboilliltoywoedf:introducing APFO contamination nto samples the Q Avoid polyttativcoethyene (PTFE), Q Avoid aluminum foil. Q Donotuseself-msemtoinoctesk. Q Avoid blue ice (the OVS tubes are shipped at ambient temperature). Q Avoid pre-viapped foods or suacks. Q Wear clothing tha has been washeda leat six ims. Q Useonly containers supplidbycontract laboratory. tFhoellzoiwpi-nogc sbaamgple collection, the OVS tubes are capped and placed ina zipJoc bag. Sel saCpomrmpopllceedtbteargt.oheugsahmtpthlbeleab.elTahnedcchoamipnl-eotfedcussatmopdlye(ClaObCel) ifoartma,cihen{0ctohltuheu tvosdluiomiefdonhefea.gir fPoarcmk.the samples in: small box or cooler with bubble wrap. Include the completed COC Complete mailing labelandaiebill andshipbyovernightcarr(0: Clayton Group Services 22345 Roethel Drive Novi, MI 46375 Contactatthe laboratoryisKarenCoonan(ERED SampleCssEhouldbe toedandshippedrefrigerated to thelaboratory. `CompanySantized. DossnotcontainTSCACBE 000311 X) Appendix-9 December 4, 2003 RMeaseyarAc.hKFaeilslero,wPLD. DCourppoonrtatCeenCternatleRrefsoeaArncahl&ytiDcealveSleorvpimceenst EDxUpePrOimNeTntal Station WP.iOl.liBnogxto8n0,3D02E 19880-0302 : CCllaayyttoonn PWraorpoksOarldNeor.N0o3.D0E3T0L10A0B8O0IS REVI Dear Dr Kaiser: pf`roWoleplaoorsweaeldpidfaaotrseetddheJtuomlepyrt1oh5vo,id2d0vea0tl3ih.deaftoilolnofwoirnSg EDrSaGfStGrEeSpoSrStSoSnStNheSsNaSpSlNiNnSga.ndThsinasldytriocsrlempeortthoisdbcaosnedditoinonosur >' p"rTheasammpslinugplairnameterssmeapycahanvgeeabsewneplroygrevsswieththemethodvalidation. Notll ofthe you have any questions, leas call me at (248) 344-1770. Sincerely, PKreomgrCahmaMrarnoangerSpecial Projets Laboratory Services |! EKncclosure 000312 K ETe-- )CwlCalavatyotnrotn CClayotonvGrooupnSeGrvicres,oIncw. Servisstae. pBmN vpmBreEEEmme Iseoi =FToPeerrfnluorop Spmoi cteaenRoic Ace idaS(nPFOHA) J r EEanan, e ns pr t PC hi fcE rR1e atseett, SO ---- D --CS TS=A iRmA--CcTRR tE--SeI--r0.r3.--1m--ea--TRe den--tm 0--3A5--e--i---- | Shipping: `Samplescanbeshippedat ambienttemperature Sample Stability:60 daysatsefioragmbieentrtetmpereaturdes Equipment ||i | 6. 7. of ter 4point 2 co td 4mLamberglassvialwithscrewcapandpolytetrafiuorethylene/rulibnbeerr.`Thepolytetrafluorethyleneside Ts ple Waters totalrecovery glassautosamplervialswithpolytetrafluorethylene/rubber liner. The | oD) S rsm 8. 5. DSyirginageb:le10g,a2s5, o5s0t&,ie1,0160uL 2 isc Emironmenta Series + Occupational Health and Sve + Laboratory Seniess 000313 | ) I D em CleWaorkyOndter:0o307e 0n834 Perfluorooe Scetatnp Aoclied ine e Clayton -- e Feweins e eteRgeo eeeeaed agenedesSeS tnasnsdtmartt sdgseiu e gedid lotew emees emess m--n 1. PefAoc (PrFOA)oAdco171s4683a,LoiO47c0MA 2. 126CPerorocteicAce (*C2FOA)nealStandard, pon, te ofysisTay 2,2003 5 Var(50) Dist 4 Methanol QMeOR),HPLCrade,Ba Ct. 4230.4 5. Ammonium Aces, Resgntgrade,LT.BakoCat #0555.08 6 2mMArmoniteAcest:isolve 0138 Amon Aciein 500ml 0 7. Exton Sovak 10K MeOH 5.DCStiockolA5e09n mn"gS3.6a5eds 0aamrnpnSo.y ofFoRnOAsS(ASc):cdfo mailpari)in 10 lofMOTT DT ion ofe --SASinMOHsuingi PFOAconcctrnion of 11.4,284,45.5, 569, 16, and 227n/a. [Er ---- DCissoolmve5e6.n8me2g3n605.i01pmoogno.fPFOA(correctedformaterialpurity)in 10mLof MeOH 11. SDpiikoinnSta5d5e5 SMoeiOnHs:resini PFOAconsetaios f 56 0d68pf. 12. SocknersSandrdSlt: CDoncientsra7ts.io2no w:g~4l702v .01mpegg/m1l.,2-di-"C-PeAcridf(clorureoctredfooromactetriaalpnuroitiy)icn 10mLof MeOH. 13.DILiniutb!eSolftheeSdoScckinoenr:sAdSadnodecSeiolninaMaeOHretdosnpln+ FrCioPbRoOsN lssoniof LB) rere 000314 | HD | Srmveomoner te ClWaorkyOvetr: 3o070n834 PerfluorooctanAcciicd 3Claytori e --= ew -- s| Qa `Technique: HighPeformanceLiquid Chromatography(HPLC)coupledwithTandemMassSpectrometry oSerEEBfcmnSrroE:nioEEpTpvT a oEmSwi EReeT--mSr E--r EEPEa t oo--ernS-- m,-- edr Esd --ix8 0 90 10 - E iE EE [Eee 9 10 90 - Capillary Voltage(kV): ES-lon 15 Sseimatasaiss w iSi Ek Eeeer Entrance(V): 0 | tEBR ocramsossroR ee1y eS ----: -- AnalyteSpecificSettings: Transition(atu)__Dwell 5) ConeVoltage(V) CollisionEnergy(eV) | ) Ce = 000315 ClaytonWork Order: 03070834 PerfluorooetanAociied yCayton &) Sampling | Supers(SKC26-30-16) asecivedfomthemame (SKC) 2. Caliatecachsnping pupwith egreseastivolctiondevice nie. 3. Dependingontheapplication,thessr twoflow tepins: (1)chFaorisnesasopxplersvonoalmuontofr4i9n0,1.sample ta flow te of 1.0Linforupto490mitesto (2)SaFomrpfleevncliangnmoonfiTtoOriLng:,sapataflow teof 15Linfor pto 430minesto bins 4. Lubethosamplewithsppropeistesampleienifcton. 5. Aftytecompletionofsampling,ocalacachpumpwith rpreseativcollection device 6 Copthecadsofcchsample. 7. Sumplsaosoredatambient pers. 5. ATchasibnoofcrufsoairostdnyrfloy.rmisfiledotadsetwithtesamplesunderambientcodions oe D `SamplePreparation Not: Samplesr extractedjstpri tosnl 1. dtnitscapcghOioVssStessaatplblieng ob int erstesions,Placeesc scionspre SSSeecettiieonn12:::XXGiIAAFDDF..nl22BeFaoxcntkporroionnooffmmeeddiiapas foplm . ` 2. ASdtdatd-m2cyocflMeepthrasneoclonod.cctesto ndhk oe6iusan echnicalfed sake 3. TGeolnmaen a1p3pmronxiGmRaEtPelAycrmisL.ofEstchsexhiocnt10bos4omrLeadmatbergerestevalebey lcetistr(o2u1g0h60) 4 1F0oWrLLoOfMaSI1M6SsamyLsteanntsfand2e0r0s.ocuaicohns(e1m3e-c0 tCsPetcotholraosossctanoosesAmpcl)er.vial. Add [) sherri | 000316 1 "Clayton WorkOde:03070834 PerfluoraoctanAocied Clayton ) |~~ I || | ! (Calibration and Quality Control 1 Cured wit origsandsover herangeof 11.410 22 fn.fo BOA. a) Calibrationstandardsarepreparedbytransferring200kL eachPFOAstandardsolutionto aglassautosampvliealr.Add 10 iL of a 1.4pg/mLinternalstandardsolution (1,2-di-"C-PerfluorooctaAcniodi)c. oi don ofan a Coming Carin Vesa (COV)Sambnds b) Analyzestandardstogetherwithsamplesandblanks.Acalicubrvr eisaanatlyziedaottn he `analyzed aminimumof everyten samples. Accce ritep riaft oraa CCVnisc +20e%. Goecnrtdanatrealssoedeiinn, vtgio1nXivfhoint.hTsosheedipnkscouat epons(3 forthecalibrationcurvemustbe > 0.992 (R*>0.985). 2 DetezminetheExtractionEfficiency(EE)atleastoncewitheachlotofsamplers. Prepaatlreaest onelevelinduplicatewithintheexpectedsampling range.Includeonemediablankperlot. 1) ct amountsaf PFOAsii snd atonote GFFportionof n OVSab. 1) Allowthetiotidyataintsemper ) Capthetubeandstoreatrefrigeratedtemperatures(2to 6 C)untilextraction. 0)Eatth PFOApk ndblk be1 eseinSleParton.Avysn `lalaiquottfraomepacehsvaram,plAeadlon1g0withosftaa1nd4ardgs.LTr.annsfemr 20S0uuLdeaachseoxtoranctsolutionto a (1,2-di-C-PerfluorooctaAnciodi)c. ) Cla aleidtTtho teecotFooFOsce RhtfoosngdfooufnPdEOaAohnodf ntheemeetcaitmteebshmoerrt aamoounnt5 ohesathi EfoPtEheOcNoveryofPFOAfs hei.Thsmean veyofreplicates `Measurement 1 `Soentptahgeeli3,qouirdcwhirtohsmuaittoagbrlaepadhjaunsdtmmeanstsstspoeocpttriommiezteecrhcroonmdiattioognrsaipnhaycacnodrdsaenuscietwviittyh.thosespecified 2 otthepied itofxhsndd odsel. 3. RecordthepeakarearesponsefactorforPFOAandIntemalStandardpeaks.The observed `retentiontimeis6.4 minutesforbothPFOAand "C-PFOA. BD I 000317 | } ClaytonWorkOrder 03070834 PerfluorooctanAcoifde " yClayton Calculations 1 Colettetheresponse ctor(RE) ox PFOA RE = (PFOAAwa/"CPFAOtA) 2 DveetrescmPiOntAhecmoansesnt(a3t)oonf.FFOAcollect usin istnde inearrgresioncurveplotingRF 9) Anyfound: ng = (sgn xEV) Where: sgn. = cbisnedbyinterpolation of bePFOApeckRFonthe calibrationcurve FV = theExtFainaclvotlumoe unl) ) Con(c3e)t= faamte Kiin)nIV Where: tas = 1 movatfound themaple (og ound/1000g/g) bo = bgamountfoundinte lanksumple og ound 1000 gle) Vo = sirsmplevoum0)e Thisreportpreparedby: :aKa+B. LSuapbeorvritsrory,SMxevtibocdsValiains Ne) SeVvicoiPreos0id,ren5t 70 Dic,Laboratory Services | February 8,2000 | BD) EEADRCTAF6oTf6 Desi 000318 $' D i |! BD Clayton Group Services, Inc. f -- p SE (ioSCASsmNumbYYeree:ee `i a sige pegr---- S SESsGHiD -- TfSigorno: ----e------ won an m r Te D e s R `Shipping: `Samplesshouldbeshippedatrefrigerated temperatures (2-6 C) pro ------ te 8 Cp S----------So--Cips AE ame A a SmseseTn TL Cl pn SAIS A rt. TE 5. Aptrd mE, Ta S IT T iTaN nal ofthelinri positionedinthecaptowardsthesample 5 13. Disposableglasstesttube, 16 x 125mmwithcaps TRGORAFT Ted Denese 000319 | ClaytonWork Onder: 03070534 WEENIE Sr rtrne:Technique: ton tvorrte--tm ----err it HighPerformanceLiquidChromatography(HPLC)coupledwithTandemMassSpectrometry E ieccmE oe--m --m--t--Sm--er--en------ EcgpEaglmnee 2c p E BaE oee sne o to ' rErrn E gEBRtmwETooee er fem ae - Pi9 E810 R3%90 oim mme --f[A EE--eER meEeL1 CapillaryVoltage(kV): 1s ee Sse mSS=maeeeilm : | |I-- MassSpectrometer Method -- . FEET m E oEER iec irpe--i t--s sis y HQ) 05 10 10 InternalStandard ENN WN (0) 0.5 10 10 (Q)=Tranussedfiortquaintiotatnion 12/04/03 DRAFT 20f4 TT LaboratoryServices 000320 | |? ||| | | 5; ClaytonWorkOder 03070634 IEEE. `Sampling 1 Samplers (SKC 226.3016) rereceive fromthemanufacurer(SKC). 2. Calibratecachsamplingpupwith epresntvecaldelvicinoen. 3. Dependingonthe aplication, eresr twoflowrtoptions: (1Foorbsicaaosapmopsleervsolummoenoitfor96i0n1g. pest flow ste of 1.0Lin fr up 1450minutetso @)Faocrmepnlecvlioerhoomtfao2r1im6n0g0es1a.mpleat lwrteof 5 Limi forup 01440min0ebsi 4 Labelthe samplewithsppoprisesampledenificaton. 5. Afiunet,hcompletionofsapling, <alibrtecachpumpwith rpeescntivcollectiondevice i 6. Caopdsotfcacbhsampele 7. Samplesaresoredatefgertedtemperatures(210 6 C). 6 hAcehLaabionroaftocruystfoodryanfaoluynsisfilledotsndsetwiththesamplesunde eiprated conditionsto `Sample Preparation Mote Samples areextractedjst prio alysis 1 EdxrictsepacoghaOssVaSebssalupbleei.ng ube i re separatesections. Placeexch ection naspare SSecetico2In.:tGXFAiFDF-oi2lFnerrontportionofmdis pusfo pl Section3: XAD-2Backportofimeodnia 2 Atadt2mcoyfMceptehleasneeoclonsod.cachtesttbe ndshakefor 0minutes ona schaicalfedshaker 3. GTealmmeanr1s3pmparmovGimEtlAyc3r.5daie..ofExctarcachtesxtsrahct o0b:3u.stlmorLdedarmebfreirgegraatsedvitelmbpelriatneg(2 r1o.u6g0h)a 4. FBolrofLO1M4SIkMgS/amn.aIlntyer,matlaSsadtaer2ds0o0fociaocnh(sampQleeUxtracEto EglassaNsaSmp vil. Add 10 Calibration and Quality Control 1 Calibratedaily withsixworkingsandads ov therangeof 11.41227ng. fr RL. 2) CaslhiesbrasttiooncsasndmaprvdlisaelaeApdre1pa0reiLd.boyf8an1s4eruingg.2I0n0tpeL.lcSatcahnWdarDdsoslautnidonrd(sGoEoEnSEos ----) TREAT Se iene 000321 | B 8 Clayton Work Onder: 370036 1b) Aegvianyiangsaanndducdosdtfogectahcehrrwui,thssdamCpolnetsinsudinbgaCnaklsb.atAcioanrVteorintcounrv(eCsCVas)alsyzedd st abe lye miofneveryi ehspml.Adepatterafno cCCeV = 20%. ) G{aectnorestanadsttanrdaarld csoacncdaetdreissc.alation sve Fon he observed pereasrespk 2. DoentenTemeilnintdheElxmcctiwoniEfifitcihenxcpyc(tEeE)d asafmpsltinogncaegwei.thIexcchh loonfesamleess.baPkreppraoret.a est 5) nject iL. amounts ofQRspiking standard solution onto the GFFpertionofan OVS tube. 1) Allow thetbe osesdtmeicys tempers. ) Capt be ndstore refrigerated pets (216C) nl extction. 9) ElxutcotohemPeRaOcAhesplikeedalnodnblvinikthbssdaddessc,rTirbeodeia2S0a0mpLleePsrepareaxtitocn.tsAoslliyozne0a3n sssuasmplveiarl Add 10.of 1.4ppm. erm Sindeeslain (REE HE. ) Csapilkeed.tTththeecronactaotroarttihonsooufdMeEchdof ohenreeosecetiosnsmus0tibeehsuomsmeedl0aamoisnthe total amountoWf EI found. Theratoels herecoveryof fom hemedia. Theme recoveryofhe pices qual the EEforWR. Measurement 1 `Saetntheali,quoid wcihlrhomsauittoagbrlaepahjasndimmaesnsssopeactproimestercchoonmdistoiognrsipnhayccnodrdasnecenwiit.hthose specified 2. thespecie lqot fcc sandardadspl. 3. RecordthepeakarearesponsefactorforWM andInternalStandardpeaks. Theobserved. retention fimeis 6.4minutesforbothJRand InternalStandard. lo | TORR Aa Games 000322 | QUPIND Appendix-10 & AirMonitoDartaiFnorgm = Sm TE ] So -- a Lrlp Ech = i 2 PPE -RespiratorAir Supplied Full Face Air Purifying wioviacidgas/high Efficiency Particulate Filter, Titan ool Medlin: __NettVast - = I [rom JRw [mr [ee i il [TT viremia TREE Ce e o eaea r nr [oEese S[o me WB) mmmmm--m---- 000323 &) EAYETEVILLEWIONRFKOSRPMEARTSIOONNNEWLOARTKRSMHOENEITTORINGACTIVITY Date: il Employee Name: Wark Area: Sample Number: Pump Number: Time 1 Comments I-- | er -- a -------- -- EE -- -- - 1 -_-- -- 7 = 1 | BH) - eae ------ TT be -------------- 000324  1 D 9 | i| | | i}| 8 Appendix - 11 Wipe Test Procedure Materials: WYPALL L40 disposable wipers; Kimberly-Clark #05701, VWR #21908-000 = Cheesecloth wipes; VWR PK-200 4X4 #21910-107 * Cotton Pads; VWR PK-100 4X4 #21902-985 Templates for Wipe Sampling 10 X 10cm paper; SKC Inc. #225-2415 = Isopropyl alcohol Omnisolv 4L 99.9%; VWR #EM-PX1834-1 * Water Omnisolv 4L; VWR #EM-WX0004-1 = Scintillation vials, 20 mL, disposable, glass w/polyethylene cap VWR #66020-326 Safety: + "The minimum PPE for this procedure s safety eyewear with sideshiclds and nitrile gloves. All generated waste is tobedisposed in an appropriate manner. Procedure: 1. Make 1 L of mopping solvent (MS) by mixing 500 mLofisopropyl alcohol (IPA) `with 500 mL of water (MILLI-Q or other water purification system water may be `substituted for EM Omnisolv water). MS is stored in a 1 L Nalgene volumetric flask. 2. Identifythe areato bewipe tested, Using the 10 X 10cm Templatefor Wipe 3. S Fillam trapntsofledrepfiiipnenettteg hewi1t0h0sMqSuafrreomcthee 1nmitn.airsecam itntoe iblleattteioe sntevrdia.l. Evenly driphalfof thecontents (~0.75 mL) on a folded cheesecloth wipe and return the balanceofthe MS to the vial. Remove the paper template and mop the test area with the wipe. Insert `wipe intothevial,gentlypushthe wipe tothebottom with thetransferpipette and : close the cap tightly. Dispose of the pipette in the waste jug. 4. Tn order to minimize the possibilityofintroducing APFO contamination into samples, the following protocol should be followed: Avoid polytetrafluoroethylene (PTFE). ADvooniodtaulsuemsienlfu-mstfiocilk.memo notes. Avoid blue ice (the samples are shipped at ambient temperature). `WAevaoridclporteh-iwnrgatphpaetdhafsobodesenorwsansahceksd.at leastsixtimes. 5. Following sample collection, the vial are capped and place in a zip-loc bag. Seal the zip-loc bag. 6. Complete the sample label .The completed sample label is attached to the outside of 7. Packthtehezispa-mlpolcebsagi.n a small boorcooxler with bubble wrap. 000325 @) t | 1| | | I ~BD : `Method of Analysis fortheDet`earnmdiPnlaatsimoanobfyPLeCrf/lMuSor/oMocStanoicAcid (PFOA) in Serum AUTHORS `Emily Decker and Paul Connolly DATEISSUED . Decomber2,2003 SPONSORS `Haskell Laboratory for DuPont Health & Environmental Sciences ~ `DuPont StudyNNeuwamrbke,r:DEDuPont-12445 3MEnvironmental Laboratory St Paul, MN 3MStudy Number: 'E03-0188 ' PERFORMINGLABORATORY `Exygen Research St3a0te58CoRlelseegea,rcPhADr1i6v8e01 METHODNUMBER `ExM-008-211 revision 1 `N TOTAUL MBOFEPAGRES wo 000326 B ) MANAGEMENTAPPROVAL |y | |i || TaeuchlnCiocnanloLlelayder, LOMS pute | Exygen Research 0 Aes fohnFlaherty pate V`iEcxeygPernesRiedseenatrch ee DSrp.onSs.oMraRrekprKeseennnteatdiyve Due . ou Dr.William K.Reagen pee `Sponsor Representative. : 3M 8) ---- Page20027 000327 ) . < | : `TABLE OF CONTENTS TETEORFCTONTERNTSBr ROrr rV r AL a nnddh fErOCHIc GURES----r-- r-- TI ALSAND SUPPLIES cm ol 34. SOLUTIONS.o..nS OFSTS ANANDDFOARTIRFICDATISON SOLUTIONS A 1 TD TT ERANr oe r 12 RS SwornANDOPERATING CONDITIONS 12 14 PE PARAMETERS. BAe 15 ) [-- 00035s8ot ~~ &) - | | | d | | ~ ' LIST OF TABLES ale, Recovery of PFOA fom Fortifications In RabbSera fable RecoveryofPFOA from ForificuionsInRabbit Plaster 8 19 LIST OF FIGURES : Figure 1 Calibration curvefroormaPtFoOgAriamnoCofntarCoolntRraoblbi`tRaSbbeituSec rum Samplen for PFOA20b FFiigguurireet23..h=ReRpe1rpnerseesLnetnattaOitfvieDveOCChBhRrOomr logram mof 4s 05 72 0 Standard for nin PFOA in Control ms 2 2 Figu`Rraeb,bitReSperreusmenwtiatthiv~e 1Chnrgo/mmaLloofgrBaCmPoFf OCoAnc c! `r Rabbie t Sers um Fs ortified r ot 05 ppb enn _Figawriot5h.PCFalOiAbrmatdiownictahv~e1frnoormPaLFtoOgOArfaiPnmCCoofPnatFCeoO!Rnc Ataeb!RiabPbEiStTPlar sma Sramplre for:PFOA2 Figaro Figare 6. 7. Representative Representative Ch Chromalogram of 40.5280 `Standard for PFOA e in Control rsrmrenin 26 Figu`rReab8b.itRPeplraessmeantwaittihve~C1 nhg/nmoLfoIBmCPRCaoOnAttrom loRar bgbn itrPlai asmanFm orotififed st 0.r5 ep2n0Zl eb PFOA and with ~ 1 ng/mL Of PCPFOA Bug Research 0003Pa2g9el] @) ~ = D | | 1. SUMMARY This report deals method of analysis for perfuorooci"tanic acid (PFOA) in = - PsBFeyOrumAmeaaninssed.xptlorafascmtaae.drforbootmicselriuumiadndspamlpalsemahbanydplremro.teinQuparenctifpiitcaatiitoornnyinofscPcMtFSoOnIiAMtiSli)es P-oFoOmyAmepilssguhiseviednngbiysnslleeicqttuiiiddonceh2sorcofimtoaintsommgoeantpihhtsoo/drt.ianngde(SRmaMs,s sTpehcetrcohmeemical(LfCormula of Tsheerulmowaensdlpilmaistmoa.fquaniation (LLfOorQth)is meits 0h.5pdpb for PFOiAn Qiunatnetmiaflicsattainodnardi.s performed using extracted calibration sundards containing a1 pTehricsewnmtetrheco0od5v%ewrays=&ds8ea.vn5ed%laorp(edTddaebuvlsieiantIgi).ornafbbToihrtesPeForvOeuArmalailnndrpaerbrabcbiebtnitsteprrleuacmsomvsaet.r0y5T,h3e,sutoanvneddr4aalr0ld BBD tion for PFOA in rabbit plasma at 0.5, 3, and 40 ppb was 99% + 63% (Table TD. i Areprvesene tatpirveecshecarnlotimibartatocigaorlniacbmursfravtoeifrooncrPuFrPOvFAeOAinfoirraPbrbaFibtObisAtesireunrmurmaarteipslhsaosmhaiionnsFsFhwgiogruwnernesi1n2. Tignre 5. Represctaive chromatograms for PFOA in rat plasma are shown ia Figures 6to8. | | | Begpen Reseach 000330 Pagesoz? ) = msD | . 2. EXPERIMENTALCOMPOUNDS Tho structureofpefluorooctancic acid (PFOA) and the intemal standard axe . ww given below. ey= FHA ChMoelmeiscuallarNwaemigeht~~ = = perfiuorooctancic acid 414 E. oH FlOF FLFFF : . BC- PFOA Chemical Name ~~ Molecular weight == perfluorooctanoic acid,[1.24i-C) 416 Tried F FCPFET >on FF `ExygenResearch Pago 6of27 000331 9) a D . 3. CHEMICALS AND SUPPLIES 31. CHEMICALS z " Grade Source" EM Science Catalog No. JT9093-2 Chemical "Methanol (MeOH) Acetonitrile (ACN) um Acetate. HPLC HPLC `Reagent HPLC `EM Science Sigma-Aldrich EM Science AX0145-1 A330 'WX0004-1 `Ammoni OmniSolv Water 3.2. STANDARDS `Standard peduomoctmoio sd (`pPeFrOflAu)orooctanciaccid, 0.2:iC) Tot Number 21002 NA Puri)ty Ed 9635 `Source Oakwood Products' `DuPont | | FcvnenReseach 000332 Page70027 @) = | {| P || || 3:3. EQUIPMENTANDSUPPLIES analytical (Ediqsupilapymeatnetast 00001 8) `Supplier Metter . BDailsapnocsea,ble micropipets (50-100, 100-2000)' __ Drummond - FClyapsesrcAarpbipdertsopa-nind vgoulaurmdectorilcumflnas(k4smm) = vario(uYsWsuRp)pliers Keystone: (Sptaarntd#-a8l4o4n0e1d7r-o4p0-0i)n guard cartridge holder A Keystone Nalgene 122m5L-mcLleLarDHPPELnCarvrioalw-kimtouth bottles `StandardIab equipment (graduated cylinders, VWR various suppliers MdMiaBslpiPotspDariboslbpeeotsIuaIbbEelxse,cCiRcoo.n)bdoutcitcilviequTiidpsha(n1dlmiLn)g -system PPaacckkaarrdd A(pmamraty#p6r0o0t0e6i5n5p)recipitation columns, 2mL.(pert Argonaut #T1so2l0u.e2a0r0r-aTy)collection plate, | mL(part #121-5202) 12x75 mm disposable culture tubes. Argonaut As desVcWriRbed in LC/MS/MSandHPLC systems section 45. iNfotthee:ycEaqnubiveaslhenotwnmatteorpiraolsdumcaeysabtiessfuabcsttoirtyruetseudlflosr.those specified in this method Notes: 1 Irneocomrmetdnodaeevdor(itdubces,opinpett, ac)m-tiheunseaotfdiispoosnab,le labware is highly 2. HPPILFCEvioarlsPfToFrtEthoeliHcnhPeedLcCckaotnuthtaeoisnaseormslpvleoenrrtmseu(qsmuteiptnmhoeatnbntoe,lu)sifenodcl.rutdihnegprPTeFsEe-nlcienoefd 3. Ttcoinstanmeicneasnstasrbyy LC/MS/MS before use. Certain lot numbers have been 4. Ufosuedndistpoobseabunlseumiitcarbolpeipfeortsuseo.r pets to aliquot standard solutions to make 5. Tccaholneitbarmahtiyinpoanenrtsstcaa(nbedlaervcdasotreadtnbddagsseeaslmipnlese)hofmuoorltdviefibctaoetiwoinctshh.ianng1emdinuwtheenfromthtehe eslyustiieomn of PFOA. : Ergon Research 333 Page 8of27 | ) | | | fm > |= DH 34. SoLuTions () 20.15mgMofamammomnoiniuummacetwaettetaend dsioslsuotlivoinng iin's prepared by L of water. weighing . - `Nmoatyeb:epTrheepaarfoerdaemselnotniognaesdtehxearmaptiloesoisftprhoevsiodlevdenftotr ogusiodlauntceea,realmtaeirnntataiivneevd.olumes 3.5. PREPAROFASTTANIDAORDNSANDFORTIFICATION SOLUTIONS Analytical standards are used for three purposes: 1. CalibrationStandards ~Thesestandardsarepreipncaontrrolerabdbit dseetreucmtoorrursaebdbiintpthleaasnamlayasinsd. areusedtocalibratetheresponseofthe 2. cLoanbcoernattroartyioCnosnctorrorlesSppoinkedsin--g1T0hetsheeLfoLrtOifQicaatnidons10axreLuLsOuaQllaynpdraerpaeruesdeadt 10 in cdoentterromlirnaebbaintalsyetricuamlorrercaobvbeirty.plaLsambao.ratory control spikes are prepared 3. sMaamtprliexsSpaitkkesno--wTnhecsonecfeonrttriaftiicoantsi.onsMaarterpirxesppairkeeds baryesupsiekidntgoinetvoatluhaetfeiethled: effectof thesample matrix onanalyticalrecovery. "Tcohreraebcstoplruotpoervtoilounmsoefssoofluttheetsotasnodlavrednstmaarye bmaeinvtaariineeddb.ythe analystas longasthe 3.5.1. Stocksolution bPryepwaerieghiindnigvioduutal10stmocgk soofluatnialoyntsiocafl1s0t0anpdga/rmdL.(ocofrrPeFctOeAd faonrdpu*rCit-yP)FaOnAd disloulutteitoon 1(0in01m2L5w-miLthLmDePthEabnootltilens)ai1s 0t0o-bmeLsvtoolruemdeitnriacfrlefarsikg.eEraatcorhatst2ocCk 106Cand isstablefor a maximumperiodof 1 yearfromthedateof preparation. 3.5.2. FortificationSolutions a. bPryepdailruetiafnogrt1i.f0icmaLtioonfstthaenfdoratoriffdi1ca.t0iongs/omlu.ti(o1n0d0e0sncgri/bmeLd)fionr3.P5.F1OtAo 100mL with scetonitile in a volumetric flask. b. Prepare a fortification standard of 1.0 pg/ml for "C-PFOA by dmiLlutwiintgh 1ac.e0tomnLitorfiltehinea"vCo-luPmFetOrAicsfollaustki.on described in 3.5.1 to 100 . Pbryedpialruetainfgor1t0i.f0icmaLtioonfsttahendfaorrtdiofifca0t.io1npsgol/umtiLon(1d0e0scnribged/info3mr.P5L.F2O)tAo 100mLwith acetonitrile in a volumetric flask. d. Prepare a fortification standardof0.005 pg/mL for PFOA containing 0.01 pg/mL of *C-PFOA by diluting 5.0 mL of the PFOA solution Exygen Research 000334 Page9or27 &) oY a | | d3es:c5ri2betdoi1n003.m5.2L. waitnhda0c.e1tmoniLtroifletihnea"vCo-luPmFeOtAriscolfultasiko.n describiend . Prepare a fortification standardof 0.002 pg/mL for PFOA containing 0.001 pg/mLof "C-PFOA by diluting 20 mLofthe PFOAsolution . d3es5c.r2i.betdoi1n030.5m.2L. wiatnhda0c.e1tomniLtroiflethine2'vCo-lPumFeOtAriscolfulatsiko.n describedin "7 . Prepare afortification stanodfa0.r00d1 pg/mL. for PFOA containing 0.001 pg/mL of *C-PFOA described in 3.5.2.c and 0.1 by diluting 1.0 mL of the. PFOA solution mL of the "C-PFOA solution described in P3r.e5p2a.retao f1o0r0timfiLcatwiiotnhsatcaentdoanridtorifle0i.n00a0v5olpugm/emtLr.icfofrlaPskF.OA containing 03d.e50s0c21r)itpbegod/1im0nL03.mo5f.L2".wCi-atnhPdaF0O.cA1mebLytodiflouittnhinenagv"io0Cl.-5utmPmeFrtLOrAiiocfsfoltllhauesetikPo.Fn OdeAscsroilbuetdioinn h. Prepare a fortification standardof 0.0001 pg/mL for PFOA containing 0d.e0s0c1ribpegd/imnL3.o5f.2*.Cc -anPdF0O.A1 mbyLdoiflutthieng*0C.1-mPFLOoAfsotlhuetiPonFOdeAscsroilbuetdioinn i' P3r:e5p2ar)etoa100fomrtLifiwciattihoancesttoannidtairlde inofa v0o.lu0m0e0t0r5icpfgla/smkl.. for PFOA . PcsooFlnutOtaAiionnisnoglduet0si.co0rn0i1bdeedpsgcir/nimbLe3d.ofi5n2C3-.5tPo.F2.O10Aa0ndbmy0L.d1ilwmuittLihngosf0c.et0th5oenimt"iLCl-eoPfiFntOhAea j. volumetric flask. Prepare a fortification standard of 0.001 pg/mL of 3CPFOA by `dimlLutwiintgh0a.c1emtoLniotrfiltehien 'a*vCo-luPmFeOtAriscoflluatsiko.n described in 3.5.2 to 100 oStttolreesa)lal tfor2tiCfitcaoti6onCsftoanrdaardmasoxliutmiuonmspienraiordeforfige1ryateoarr(finro1m2t5h-medLaLtDePoEf preparation,aftrwhichtimeitisnecessary to makenewstandardsusing the stock solution. | | ExygenResearch 600335 Page 100127  = D | | | ! ) 353.Calibrati"nToihneSctcaaallnidbra0artdiostnhestasnadmaprldess.aro Tprhoecesfsoerdiitcharioougnh otfhetehxetrascttainodnarpdrsocebdeufroer,e _ extraction doneaccordtiotnheg following tbl:AB os CMoixnedOf VFoorltuifmieca(tiio)n Vo_Clounmweolof Ctornacc.teofd FoSrtoilfuitciaotinon Sample (mL) CaSltiabnrdaatridon 00015 E50n0 000055 01s 100s 550000 000055 150 5200 5500 000055 250 4. METHOD T4h.1e. FflroowwdDiuagsrcaRmAoMfthemethodis given below, followedby adetsled descriptionofcach scp. Mea0.0s5minLtuoop`frrsoaeMminepe lpFerlet (couiwpsDih itniagatodigiornsdpacomoslaublmenmicropipete) 4 tein piptaion n+cn ingMolto LC/MS/1MS analysis 42. SAMPLEPROCESING SSNaaommppsllaeemssplmesutsportroecdbeesrseaiflnrlgiogiwesrenadeteetddoescdhofomouprlldesteearlsyuomthobera.wp,laaluslnmo-awiedsdeadm,tpolaetseq.7ui0lH0iobmwreattveeemrpt,oefrrraootzoee.mn temperature. All samples must be thoroughly mixed before being sampled for extraction. Bxypn Research 000336 Pugs 110f27 = | | || | me o || | || }| J : 4+3.BSaAcMhPbLaEcPhRoEfPsAaRmApTleIssONanoalyzbedan()yp,icalnleym35aoir1ecso9nFicwmeintchlouddebaltalnoekac)se,tduo0rnael +_ ETreeacgotevnetrysCforrotalhmepoblaietceihcbdatcah1 shJowuolwdaboccoxnttceedrinn s-u-p.1U0ERvCtey LpErTown A b A eeretiindesicakemxmiaeacdyibetdoopushrgofhuoldmtehbedoapsehrpoasscploeynsorr0'is efvbeelstrecovery. 4+4.CoEnxfTiRgAuCtTrhIeeOMNuliprob deck withthe folowing iDWer: 22 1.11203007L5mdmistpeosstwaibblheepr9o5acnk recipation columns ad 1 mL colecion rf L4.eT(ipplucghauontyefuintuhssedaidmsphpoollseaslimobiinaiegthl:ee:acSpoPnpEtramoianpnirinfigorlhdoe)tspeprBoprEiaEeco)ncceonltruamtnio.n of a PBlaeceeatalnleadispntr1etoaheblfeo'slulsomwdciaonrngdosnltteospof8adtiwa3srpe0o,sWa0ibnl:per1e2p,x7e5imsmters OuBbe=.that L1.O8fGetoclsean pyip5e0t0euipvl(eso)f.1A2C7N5msomluttiosn fobleltow)esdnbdyarsainsctBhoaereSPaaEirDmpleu8gr. le 4l.an0de tdoritsehpseebbntsyiepcthhueteAaCneNd dsirolounpttihponce.itp(iSa)t-ionclolAuCmNns) unshavebeen. bReee mnalilprreqbuiormepdarporncoeitduirnveapsrceeucguispmhio1tsa-yut4sigutohennmchoiolnup1mr0no1st0.ei2n0psreceocni7o,Scoulnutmiln A only 1i01o0n1a3 cheayusemsb)e.eondNodetroaefw:nhAdeiWoinnparle vmaactuhuomd0ndsslinbge i1nitiated evh acuumo step "Praenviaew.o" savismitph lpipeet cfo aly: . 45. QUANTITATION I4n5s.t1r,umeLntC:MSMS Sys"BtPeaEmSeaCnkIdmEOXpaAeorPaItli4on0ng0SC0porBnaidyiotmLioiolqneusciudlIanrtrMoadsucstiAonnilJytaeeftice Computer: Dell OptiPlex GX 110 000337 BogenReseach Page 12612] ) = | JengD z i|i |! BD) Software: PESciex Analyst 12 `HPLCEquipment: He`wQlueatttpPumpaGcI3(k1HP1)aASerrides 1100 a ,- Vacuum DegasserGI3224 = ACuotlouinmjnecCtoomrpGaIr3t1me3nAt GI316A NSoetee:io7T.w4p0oon0t)ta4orexar1ta0tpaamcnhmyeTd ishndy-plueiernaecatofdnetdaromtpihnioannptugsrughaaertvdmaciaavryeibaidengdienbe(tfhKeoermeyotabhniel,sepipemhtpislee aHnPdlLoCrCHoPlLuCmns:ystem.GenesisCp GonesChromtogeaphy), 2-1 mmx S0mm4, CIonljeucmtinoTneVmopleurmaet:ure: MobilePhase (A): 30 C 52mHLMAmmoniumAcetate in Water `Mobile Phase (B): Methanol Tine 2A %B Flow. (mLimin) 030 3s "0 @ 100 0033 03 3577 00 110000 0035 03 955 2 "0 @ 0053 03 ItmaybenecessarytoadjusttheHPLCgradientinoder ooptinize insiurient performance. Ionsmonitored: PFOA Negative BCPFOA Negative anstion Mi . 413-5369 415-5370 Approximate 28 min. 2.8 min. The reetTenwtiromon,t2pepshliaonsmgeal,yacsecvta.hreyD,iroiinfftaciodnnatyir-netuoee-nsdtaityohnbotauisgimshe,stdiheespeeannctdciirenepgtaaonbanllyetshwieistcahonildnumtnah,e Sotmadnidayridaslare interspersed throughotuhet analytical run. Note:Analternative LC/MS/MS systemmay beusedonce,d- emonstzatedtobe equivalent. ExvaeaResearch 400338 Page 130027  "4STo5hle2um.tiaosnosTfsunPepFFOieAle(cPaatrr1ai0mseoittLnemmerisdnef,orutseiaencghanainnfusaiboynilnpfuyumspit)ngvaei-2a 1"pTg"/nmLf.osatasntdraearmd ~: etooeeant t0.ao3nheamdsLef/ocmroitnnhteafpilnroiowndgrua6ct0et,%foTmn.hetehOaannncaoelltythaeenidisni4sn0itt%irao2llmymesMnusnatemudfnmeoodrn,ittuhhemeoppaactreietmnattiezieoidnn paraar msaveed ts aetnrefisle. This tun file isthenusedduringroutine " analysis. 45.3. CalibrationProcedures o Jojectthe same.aliquot (5 KL) ofeach calivration standard ino the LOMSIMS. b. UtSsieevlmei.eneraprfseosattkaaonscddrahoraadfcmuenrmvabelaysflsoitlnraenadrqyaruraendgtrtieptsaestaieiokonn.aurLseiianngevUaerrxssuwstentdihagerhdsctaotuifironvthegoesfraattrhieoe | OCOoerimsataaecuanilvtaooultnelni)oefrbsaoynfatluwysatirenelgcstoyhnsecteaenptmpr.rao_tpiAroinnayotfecoiaunlttilebirmeaartliteossntta(snetd.aagn,rddFaruudsgiefngoOuuAntnldailetyrosTtbees1?.)a2 | ~~ naybrrbaetieoaxntsctlsautndadenaddrafdrrsdtsohmaitntjcheeocutcleadldibbarenadteixoacntlculuderaevsdet.mounsHetownceoavtleeibrr,axttihocn2os0ett%aalnoednfaurdtdmhbeaettrottohafle |s LeLOQ must be retained. . T2Zhp0oe9r9oc2po5tmsell(eaRt?iso2ten0p9s8co5me)uf.sictiIebFnetcat(laiRbk)reanftoitrooncaraedlsijubulrstatstiifoannlsltcruourumtvesenistdegoeptnehererastaeitoelndi,mimtausns,dtthtbehene daorrthd ereles vantstofsamplesshouldbe reanalyzed. | 4.5.4. SampleAnalysis a Injtheescamte aliquot (5 uL)of eachstandard, sumple, recovery,contro, ic ino the LCIMS/MS system. | b. Standards corresponding to at least five ormore concentration levels must be included in an analytical set. c. `AASencoefunontlitlrfoeowsreedtaosbfeyccaoclanilbdirsbaretatitooinofnsststatanandndadarardrdsdsss).ihnAtesorbsaopiunenrasjleedlctdeetvdereayt mt5ha-ea1nb0eetngstiaiinrmnepivlnsegcestoo(,f21f0 SSealmibprlaetisoents.taInndaeridthsemracyasbe,ecailnicblruadteidoan tstthanedbaergdisnmnuisntg abnedtahte tfhirestennddoflasat injection in a sample sel BD 4. TToehnsepdocnoasnecsencstuhrrovauetl,idonbbarosafecckdeatcohznessatpmhopenlsepe/fesoaorktfitfahireceaarteisooifnd/ucceoanfctohruolnadniasilndyteeeta,ecrhmsiTanhmeepdlfesrtsaoenm.datrhde Boygen Research Page 14of27 000339 5 Jip 3 D : || | If necessary, dilute the samples in 50:50 methanol: water containing 1 ng/mL. of PC-PROA to gi a revspoe nsewithinthestancudrvaerrandge e. tFhoertLificaLtioanrOereccoonQ vseirdiee)rsefdelalcicnegptwaibtlhei.n 85 to 115% (--80 to 120% for levels at ~~ f. Extracted samplesmustbestoredrefrigeratedbetween analysis. 2Cto 6Cuntil . & cSoarmrpelsepsonidniwnhgiacnhalnyoteperaetkesntairoendteitmeecstewdil(li.b.esrigenpaolrt:endoaisseNrDati(on<ot3d:e1t)ecattetdh)e. `tSiammepslbesutinarwehliecssh tpheaanksthaerelodweetsetctceodncaetnttrheatcioonrroefsptohnedicanlgibarnaatliyotne srteatnednatridosn (0.5 ppb whereLLOQ = 0.5ppb)willbereportedas< 0.5 ppb. 46. ACCEPTANCE CRITERIA TPhFeOAf:ollowing criteria must be met to ensure the presence of PFOA and C- 1. Thechromatogramsmustshowthe following peaks: PFOA: a daghter ionat369amufrom aparentof 413amu 13C.PFOA: a daughter ionat 370 amufrom apareofn41t5 amu 2. tAhneyLLanOaQl.gte Aprneyseanntalinytmeeptrheosedntbliannktshemruesatgebnet abtlalenakstmu3s-tolbdelaotwleerasttha5nfoldlower thantheLLOQ. 3. R115e% c(80o-o1fv2l0a%ebfcoorrntlreiovleslespiakstesthaenLdLmOatQr)ixosfpithkeeisrmkunsotwbnevbaeltuwese.en8A5n-y lmiemtithsodwarfroartnitfsictaet-ieoxntr(alcatbioncoonfttrhoelenstpiikree)anafallyltiincgalousetts.idAentyhemaatcrciexptsapbilkee outside the acceptable range should be cvalusted by the analyst to determineifre-extraction is warranted. 4. Aanppyrocparliiabtreatoiuotnlisetratnedsta,rdmfaoyunbedtexocbluedeadstfartoimstitchaelocuurtvlei.erbHoywuesvienr,g tahne extoctealednu2m0b%erofotfhcealtiobtraatlionnumsbtearndoafrdssttahnadtarcdosuilndjebceteedxacnldudaetdlmeaussttonnoet calibrationstandarda theLLOQmustberetained. : 5. `The correlation coefficient (R) for calibration curves generated must be 2a0pp.r9o9p2r5ia(tRe?s2t0e.p9s85m)u.stIfbcealtiabkreantitoon ardejsuulsttsifnaslltrouumtesnitdeoptehreasteiolni,mitasn,dthtehne standardsor therelevantsetof samples shboerueanallyzded. styeson s00o0n340 ) ~ = me D | | | || | || | 4.7.PERFORMANCE CRITERIA t`hTehecofmolmleonwciengmetnwto corfitaenriaalymsuiss,twbheenpeursfionrgmeadn isnss2tsryusmetnetmatisouintasbeitl-iutpythteast,habesfonroet been usfoerthdismethod. . k aRnudnobatasitnanadsarid gsolnutaionlo-nratLtCioo/oM-fSan/tMloeSaisctos9r1ree,spcondoingmtotpothaeraeesatgriemnattebeldadLnkL.OQIf tphairsamectrietresr.ion. cannot be met, optimize and change instrument. operating cr RstuarntiangasettoorfbsetlaonwdtarhdesLoLfOfiQvueportomtoherehicgohnecsetnctornacteinotnralteivoenlls,evseplatnonbinegairncalnudgeed in the analysis. Generate a calibration curve for the analyte and obiain a linear rcergitreersisoinoinsmweitt,h saacmopelfefsicmiaenytboefandeatleyrzmeindawtiitohnst(aRn?d)aorfdsatinlteearsstp0er.s9e8d5.. Once this 48. TOMEREQTOURAINARLYESIDS + lAasbeortaotfor3y5csoantmrpollesspi(k1esr,eaagnednt3c0onstarmopll,es1m)actarnibxeconttarkoenl,th1lraoubgohratthoeryesxptirakcet,io2n p(rsotcaenddaurrdesiannadp3p5rosxaimmpalteesl)wy i2lhtoaukresapbpyornoexpeirmsoa9nt.hoeu`lrTshy.eLC/MS/MS analysis 5. CALCULATIONS a. pUesaekBqauraetai)uosni1ngtotchaelsctualnadtaetrhdecaumroveu(nltionefaarnraelgyrteessfioounnpda(rianmentge/rmsL),gbeanseerdatoend bytheMass Lynx software program. Equatio1n: `Analytefound(ppb) =(AnalytePenkarea peakaca).-Intercept)xISconc.(ppt) . b. FEoqruastaimopnl2estofocrtaifliedcwtuihtehlpkearnctoewnent aremcoouvnetrsy.of analyte prior to extraction, use REecqouvaetriyo2n(.%)= . Analytefound(ppb)=avg.A`nAamlovuenftoAnuanldyitneathdedceodrr(epsppbo)ndingsample.(ppb) x 100% Bogen Research 000341 Page 160127 i | 6. SAFETY aogeeantsy`bsefohreopderfoadrmfineg tthisimlethiodt:y dUasme hunisversaolr p4recO autioS ns when + -_ rm Ee `nanddgloovges. wUrsei`tbloof d-beomebpoathogen`handling lparbeocraauttoiroynscwohatesn, shaafnetdyingglassseersu,m and plasma. | -- : 000342 purer ) ) I=. |e D || | || Table L. Recovery ofPFOAfrom FortificationsInRabbit Serum `SummaryofPOA Recoveries for 0.5 ppb Fortification in Rabbit Serum - BogDen TERT Spousor DBaattaecion ADnaulyesis FortotLe)vel TAPE Om oni 03 Recovery 5os GCOAO0SM00SFRoFtt32 LLoottoSoSHHBSAANN)) 0055022/0033 Res LeRRR 005520330033 003 ------maes 1 `RELATIVE`SSTTAANNDDAARRDDDDEEVVIIAATTIIOONN:: 5511 `Summaryof PFOA Recoveries for 3 ppb Fortification inRabbitSerum Bogen TERA Spoor EDimaccion DAuwmleyis FortoLnevel ToOE00 Gs Gs 3 %Recovery T1o DO08R08FFoorrtt 56 LLootto9S9HHBIAAG0O0 005/2S203 00S522330033 33 1607 -- psRa-- re m-- B w `RELATIVESSTm TAANNDDAARRDDDDEEVu VIIAATTIIOONN:: e3320e `Summaryof PFOARecoveriesfor40ppb Fortificationin RabbitSerum Exygen RTT SpDoor BDiamtceton ADwamtlyesis ForptLsevel TAM oe Ga 4 Recovery wo O0O00R0ON8SFFoot8ro LLoottooSSHHBMOO0D 0O57S20S3 00552330033 "" waa s%u ey `RELATIVESSTTAANNDDAARRDDDDEEVVIIAATTIIOONN:: 0055 OVERALL STOAVNEDRAARDLLDEAVVIEARTAIGOEN:: OVERALL RELATIVE STANDARD DEVIATION: 855 8 ExygenResearch 000343 Page 180127 ) - - ~D 3 { | | | Table IL. Recovery of PFOA from Fortifications In Rabbit Plasma `SumomfaPFrOyA Recoveries for 0.5 ppbFortification inRabbitPlasma - BayDgen Spoor BiDctwion ADmbueis Fort Leivel %Recovery T Coracs | LadiT n Gms GEM 0B 5 100 mid 0300922Fort. 3 Lot40K7400 05/22/03 ST0A5/N23D/A03RDDEVmI0AT5aIONs: RELATIVESTANDARDDEVIATION: m1o0Ws0 ow SummaryofPFOA Recoveriesfor 3 ppb FortifiniRacba bittPilaosmna ByD TT SpBoror xcCiheoen TOG Gms ADsws FontGLmew! oa 3 Recovery Tor CCoooppprtoanss LLawdiinn oowms oonRy 33 11006s S mind Ja Es RELATIVE SSTTAANNDDAARRDDDDEEVVIIAATTIIOONN:: a10 `Summaryof PFOARecoveriesfor40ppb FortificationinRabbit Plasma JBiey SpBDm BuDmucieon ADuws Fo Lowel JE Sa Recovery DE Coamprearso aLa0 dr i SosEm EenSum io a em RELATIVESSTTAANNDDAARRDD DDEEVVIIAATTIIOONN:: o6s OVERALL STAONVDEARRADLLDEAVVIEARTAIGOEN:: OVERALLRELATIVESTANDARD DEVIATION: aa . Ep Reserch 000344 Pugs 190127 ) Figure1. CalicurbvefroraPFOtAiinCoontrnolRabbitSerum nt ei OR msern080550 | / | 1| 1w8] A 2D) CEFTT ET ew BoypenReseach EEA ER SEA IEEE ER] : 000345 Paget) : ) Figur2e. ReprmepsleentfoartiPvFe OCAhrwoimtaht~o1grnagm/moLfaofCoBnCt.roPlFROaAbbitSerum `Sa ESET . Co = ERE og ii Dd = =. EFT: oo || E Be E BE J i Fuvaen Research Page210(27 000346 L 2) ) Figure 3. RPepHrOesAenitnaCtoinvteroClhrRoabmbaittoSgerraummofwait0h.5~p1p`bngS/tmaLndoafr"dCf-oPrFOA E =Es =3 CL- ~ ii 2 i . EEE a a Eide | i . | BaygenReseash Page220077 0003477 d, - Figure 4. Representative Chromatogrart, OA of Control Rabbit Serum andwith ~ 1 ng/mLof BC- Fortified at 0.5 ppb witPh F FOA . - P a Tn+ Exiz= iIE == - ] EExE.E F P ii ! FxvgenResearch Page 230f27 000348 Figure 5. Calibration curve for PFOA in Control Rabbit Plasma ref Rew TD) 4 - | ~ |i ETT R EERE TEETER BD -- nate 000349 ) Figure 6.Representative Chromatogram of a Control Rabbit Plasma [Em . Sample for PFOA with 1 ng/mL of BC.PFOA | EB D> | EponResear Page25l27 000350 ) ; Figure. Representative Chromatogram oaf0.5 ppb Standard for PPFFOOAAin Control Rabbit Plasma with + 1 niyg/mL of Be. jE hms | Ds | | Er || i BxpgenResearch | : Page260027 000351 FTigiuree8. aRte0p.r5espepnbtawtiitvhePCFhOroAmaantdogwritahm~o1f Cnogn/trmoLolfERsab*bCi-tPPFlOaAsma =) . IEE = 7 || | | | | ! t i E w= B FE=R= BEE E BBe-FHsE 3 i id `Exygen Research s wr EEEEEE . Page27of27 000352 |: | Jom D | || | | December 4,2003 RMeasreyarAc.hKFaeilselro,wPRD. `CDourppoonrtatCeenCternatlerRefsoeraArncahly&tiDcealveSleorvpimceenst DExUpePrOimNeTntal Station PW.iOl.miBnogxto8n0,3D02E 19880-0302 GCilaayyttoonn PWoopraksOarldNeor.NGo.30D30E1T00L80ABROE1VS Dear Dr. Kaiser: WIeoalroemepdleoarsteh tmotprboovdivdaltihdeatfioolnlfoowrinGgDrUaftEsporSt onSth EsumSplin.g nTdsimlyriacfarlempeortthosdbcaosnedditoinonose prodaptedolys15a,20l03 Tpahreamseatmeprlsinog uprnsoeienrtshismraeyocrhtahnagveeabseween uprloygrveaslsiwdiatthedhe method validation. Not all ofthe 1yo have anyquestions,please call mst 24) 344-1770. Sincerely, PKreogmraCmhaMmanoangerSpecial Projects Laboratory Services Exncclosure 000353 | | | LD ' : | Clayton Group Services, Inc. Eprom Molecular Weight: CAS Number: ---- NosOitSk:E NNoonreo pAE: r0t36pmb i ---- ClaytonWork OrerNo:_a3tnogss Draft ReportDate: Decem4,b2e00r3 PApopperenscs; Whi Powder MSiiengPoin:_ sm Sampling and Storage Staple: Oba VerseSample SKCCot No2263016 Stage: Tobedetermined `Shipping: Samplesshouldbeshia prefprigeeradted tempecatures Sample Sabi: Tobedetermined Equipment L. SKCOSHAVest Sam(OVpS)CearNo. 226 30-16 2.HdPeLeCrSystem:LCpupithstmtgait onl, osamplrapoaa UVbrace 1 grretme 6. MSVaisaksErSpcltomectrp:roTr,iepleegqsaatirvrponldepmiyseseroation 2de1a00payma,masndeGnug sciiaoma,heAPtLinoince arent - 6. 6mumber hssvi wih re cap ndTeleor 5 4 amberhss i with cecapandTeonbbenre 10. Water on recovery hs stovmilwpithTellmer 12. Synge:10,25,50.8 10041 13. Dispoglasssatebsttlubee, 16 x 125mm withcaps Te ee 000354 ) | D- Clayton Work Ordere: 0m30m7m08m3a4 smtPeermfslruoeroomcteanmoioceAmcid eee e ee ee r eMreaasusrs eemernt peamt opmer lee ee sas Technique: HighPerformanceLiquid Chromatography (HPLC) coupled withTandem MassSpectrometry Detectors: TripleQuadpolMaseSectontr(imsQuatreLC, civalen) Eximcton: mLofextractionsolventfor60 mints Columf: rAgiiletntPiRLydpEesCoODiSn,s2.1 x 100m, im iSpeeroler Conditions {onizaionMa: ctNer gsive CIonFletiuimoanntTVeonlgu:me: S030l3.mCite SDNeeecsbDcuGkaGlvssT(aa(ULBeeie:)o:m69n60(pC1e20 RTeoennTioomnesTine: 614 mmiineess DCetvsoioGnaTleremapro esr(smn Can3y50163 Miler 60 MobHiTOPuSs W6Gradienat am Eton Sono ow Sones Stings W9 owo1w0 2onw C- RECxaFpticLllteaorry 1(V)oV:l:tage(kV): oi1s5 BAa=no!moni Ace iNNGSSLClRMmMaBGoerRyoeOsonn0: 51154 WaveleoghUV): NotAppice EaM2neosmB(oVr:y(Vy: vis Baty o {MteSrFCuhnacnonenl:DMelaaylflorMReRsDto(n0M.1ori(iMIgD) ESP`SShparntDaoltoonn: 0.5Endtine: 14min `AI = mlytenSpSeocinftdicSetWr tingsD :a`po1To4n-93a-603m56@a59)D00l35 @ Trani ed orqt Conpei1ola 0. CalsiIn Eos(41) BE 000355 ) [D | | coeveromeomm reeleuormaicadd SamplSiangmple (SKC 2263010 me sivdfmte mac (KO. 2 Cate ch pling pap vith rset lion doris oe. 5 Depminonte pplcsion,br ew low i pons: (1 Forno penmorn,ample fwtoL0iin 0 0400mits0 oy oievlee lAigLof9a6ie a fo to 1 ino 10440 mits 0 i 4 5 Labbe plewithpop pledesifcsion nubeconminionfmpl cbr xhpupvith eprseniecolton evi [Ss PRS-- 7. Samplessesd fg pes. Achinofcuttyfos led tdsenwilh hesample de gedcondos 0 Tabooamis SamplePreparation Nets empl ecco nis.Semper bleor XdostX pers 1 EGxitoaS cpghOaVaSosaGplhRlie,ngebrontepct ection. ls ihsection 345 SSimm3:SXAADD. Focnktppironoefmmmdisph fom 2 Add mafiotto ch stn ndsakefoc 0 ies on chic ok + meTsopitAciml.ofrachneshxout.sorleademeirghns.ilby Brio gh 4 FrLOMSMSmi tirn20 iixohspectta spp il.Ad10 Calibration and Quality Control 1. Calibratedailywithsxworkingstandardsove therangeof 11.4t0 227 ng/mL.for WRENS. 0 Cagllaisbsraauttioosnasmtpalndearrvidasla.reApdrdep1a0reudLbyotfaran1s.f4eprgr/imnlg.2I0n0teprLnaelaScthaJndlardsosltuatnidoanr(dsoElutEionEto 3 irrTpeeaser----pp------p = ep re000356 I----. @ | | rE || | Clin WorkOnder: 3070834 Perfuoroactanofe Acid 1) Aegrieuyeidntm anddeni ogfceacvehem rriiy,tehnndo CppollentasnidAgbfaCspkrasa.ntAocn aVserlifficiatcibCiornOvrVe(aCCa4V.y)ts2ed0i.aoatnhe Gcenoerratseon standi cnrt . clr onsva e om het observede peak rshs espose: 2. DoeetmvintetShpBxecltoinifnfheixp(EdE)at psitngaronwigt.hoclhdetoofsendla.barPkrpepearreot st 3) Inject iLamountsofJ spikingstandardsolutionontotheGFFportionofanOVS tube. 1) Allowthotbeto idoatamitrps, Cupthe be dsr t efgrated imps nlxcion. d) Exdtroarcttohe cN-spikledeanodnblawnikltusbeosadsd.escTribeadinS0am0plePeraecpahreantiaocn.tAsnaolynzesan passautosamplervial. Add10kL of 2 1.4 ug/ml.InternalStandardsolution GREENE ------ ) Caalbcleua dla.tTehto heocroaftaicu oefnorrttoit hoennnsg.foafnJiMnRefcohunofdonthresmedeicatiponikeottothbeetmheaormeteicalacmoaunnte CThaevrsm hEflofcosrhere of WRfom heme.Themes sve of eels `Measurement 1. Seatptogee 3,o cihlromsaitboglreaphaadsmsspptttecchonodmitoigonasphsycaodneceywi.th hosespecifi 2 intospecieallotofexchsada andsample 3. Recoirodnhiehee 1s4rispsonsoe cothofo MAeRndandotoeerSaSnanhd.a pss.Theobserved TA eT mn 000357;