Document Qgb6db1BaxbOD9Qy0X7db6Na8

| ARGOS p55 | BEST COPY AVAILABLE i Laboratory Composite Report : Analytical Reports of Data for Fluorochemical Analysis in Human Sera | LIMS No. 1623 | ! | | i | | TT | 1 | | : Testing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory Fluorine Analytical2C3hEem0i9stry Team (FACT) 935 Bush Avenue, St Paul, MN 55106 Laterson Comm Kris Hansen, PhD. Bldg 2-38-09 P.0.Box3331 St Paul, MN 55133-3331 Phone; (651) 778-6018. FAX:(651) 778-6176 Requesters Larry Zo3bMelM,eJdeifcfaMlanDdepealr,tmGeenatr.y Olsen Bldg. 220 St Paul, MN f 004345 Page I [I :i M"Taiseco2m,po1s9i9t8eornedpoArgtenlu2d4,es19e9n.eEcahcihcraeproepodrotcsuametnwtesrteheirseedlootf eslSMyMedoifcbaalmDepasre tmenurtbplwees en cdelioverlyoffrltohmeevaarciotutstsheourcdes.estIenrmIontsetrcprsectsat,iocnaochftheechriecsaulrsepohrtenrcehsdpeosnsinbfolrymaotfiohne b3M Moetlhieuadaltteof | Der: PArlotcheoduugrhersiagonrdomuestqhuaoldistwyecroentfroollmloewaesduwrheesnanpdas3sbMlEen,vtihreoamcequnitsaitlioLnaboofrahtiosrdyaSttawanadsarndoOtnpetciesnsgary | hcoelllpecctheadraaccceoirzdietnghe(p0oatpepnltiicalabfloeGhouomdaLnabeoartathocroynPcearcnstsit.vDaasntaawaalsvcaoylslpeocstseidbtloemteoefotillmomwedlaibaotreanteorsyds0 pSraofc,edbuurtewsatshatowterreeviinepwlsabc yathethQeutailmiet.yDsautraawnacseteUcrhniicianllpylarceevaitew3eMd.by 3M EnvironmentalLabechmical i eee taiipit {echnicalreports | `Serum forAnalysisUsingHPLCElectrosprayMassSpesromerywiththefolowingexcepi:ly!cctate "Wasaddoeaqueousexact asicadof MTBE. I E`xwtarsaccotlslUcsitndgusRinPgL ESEMlSe,twrohsiprdaaytMaafsosrSspuebcsreqoumeenttpyh.aDsaetswaeiorctcuodleldecitedheuPshiangsEeSIaSnMdPSh.aDseetIaTilrsecpoarntbse: 1 foundincachtechnical report,includedin Appendics BtroughK. i Chromatographicdatafo cachtechnicalepisoarrchitveatthe3MEnvironmenLtaabl. ; 2 Ses OEE hE al ] rms Ee || . A `AppppeBA:n:e PAhtnaalisnyetx2icdalMi ethoxds. | ``AAppppeeCD.n:nPPddhhaaiisseexx3 ``AApppenpdeiEFx::nPPhdahisaex6s ```AApAppepnpdpeiHGIx::.nePPhPhdanhassiaeeds9x78,epirtx| | ``ApApepndpiKx:ePShuanmsmdarpiyaoxftPOAAlevelsforPhases 2.9 ! Page2 i 004346 | 3M ENVIRONMENTAL LABORATORY. | METHOD | EXTRACTION OF POTASSIUM PERFLUOROOCTANESULFONATE OR OTHER . FLUOROCHEMICAL COMPOUNDS FROM SERUM FOR ANALYSIS USING HPLC- | ELECTROSPRAY/MASS SPECTROMETRY | Method Number: ETS-8-4.1 Adoption Date: 03/01/99 1 | Author: Lisa Clemen, Glenn Langenburg Revision Date: | Approved By: ' Laboratory Manager Date | Group Leader Date ] Technical Reviewer Date ' 1.0 SCOPE AND APPLICATION 11 Scope: This method is for the extractionofpotassium perfluorooctanesulfonate (PFOS) `or other fluorochemical compounds from serum. 12 Applicable compounds: Fluorochemical surfactantsor other fluorinated compounds. 13 Matrices: Rabbit, rat, bovine, monkey, and human serum or other fluids as designated in the validation report. . 004347 Word 6/95 ExtracoEtfTSiP-Fo8O4nS.1from Serum Page lof 14 2.0 SUMMARY OF METHOD 2.1 This method describes the procedure for extracting potassium perfiuorooctanesulfonate (FOS) or other fluorochemical surfactants from serum, or other fluids, using an ion pairing reagent and methyl-fert-butyl ether (MBE). Inthis method, seven fluorochemicals were extracted: PFOS, PFOSA, PFOSAA, E{FOSE-OH, PFOSEA, M556, and surrogate: standard (see 3.0 Definitions). An ion pairing reagent is added to the sample and the analyte fon pair i partitioned into MBE. The MIBE extract is removed and put onto a } Fniilttreoregdenthervoaupogrhaato3rcucntpilladsrtyi.c sEyraicnhgeexattrtaacctheisdrteocoans0t.2itwutmedniynlo1n.0fimltLeor ifnmtoetghlaasnsola,uttohvieanls 22 These sample extracts are analyzed following method ETS-8-5.1 or other appropriate ] method. 1 3.0 DEFINITIONS 31 PFOS: perfluorooctanesulfonate (anion ofpotassium salt) CFyrSO5TM | 32 PFOSA: perfluorooctane sulfonylamide CsFirSONH: 33 PFOSAA: perfluorooctane sulfonylamido (ethyDacetate CyFirSOsN(CH:CHs)CH:CO: 34 EFOSE-OH: 2(N-ethylperfluorooctane sulfonamido)-ethyl alcohol CsF17S02N(CH,CH;)CH,CH,0H 3.5 PFOSEA: perfluorooctane sulfonyl ethylamide CsF17SO;N(CH:CHs)H : I 3.6 M556: CsF1;SO:N(H)(CH;COOH) | 37 Surrogate standard: 1H-1H-2H-2H perfluorooctane sulfonic acid 4.0 WARNINGS AND CAUTIONS | 41 Health and safety warnings 4.11 Use universal precautions, especiallylaboratorycoats,goggles, and gloves when ] `handling animal tissue, which may contain pathogens. 5.0 INTERFERENCES | 5.1 Therearenointerfkneowrneatnthicsteimse. | 60 EQUIPMENT t 6.1 aTchceepftoalblloew.ing equipment is used while performing this method. Equivalent equipment is 6.1.1 Vortex mixer, VWR, Vortex Genie 2 6.1.2 Centrifuge, Mistral 1000 or IEC i 6.1.3 Shaker, Eberbach or VWR 6.1.4 Nitrogen evaporator, Organomation 004348 d ExtractionoEfTPSF8O4S.1from Serum Page2of14 | 1 6.1.5 Balance (+ 0.100 g) 7.0 SUPPLIES AND MATERIALS 7.1 Gloves 7.2 Eppendorfor disposable pipettes 7.3 Nalgene bottles, capableof holding 250 mL and 1 L | 7.4 Volumetric flasks, glass, type A 1 7.5 I-CHEM vials, glass, 40mLglass 7.6 Centrifuge tubes, polypropylene, 15 mL 1 7.7 Labels . 7.8 Oxford Dispenser ~3.0 to 10.0 mL. | 7.9 Syringes, capable ofmeasuring 5pL to 50 pL 7.10 Graduated pipettes | 7.41 Syringes, disposable plastic, 3 cc: 7.12 Syringe filters, nylon, 0.2 pm, 25 mm | 7.13 Timer | me : 7.14 Crimp cap autovials and caps | N7.o1t5e: PCriroirmtpoerussing glassware and bottles, rinse 3 times with methanol and 3 times with . Milli-QTMwater. Rinsesyringes aminimumof 9timeswith methanol, 3rinses from 3 8.0 REAGENTS AND STANDARDS | 8.1 Type reagent grade water, Mill-QPTM or equivalent; all water usedinthis method should beMilli-QTM waterandmaybeprovidedby aMilli-Q TOCPlusTM system ] 82 Sodium hydroxide (NaOH), J.T Baker or equivalent 83 Tetrabutylammonium hydrogen sulfate(TBA), Kodak or equivalent | 84 Sodium carbonate (Na;CO3), J.T. Baker or equivalent 85 Sodium bicarbonate (NaHCO), 1 T. Baker or equivalent | 86 Methyl-T-Butyl Ether, Omnisolv, glass distilled or HPLC grade 87 Methanol, Omnisolv, glass distilled or HPLC grade 88 Serum or blood, frozen from supplier 8.9 Fluorochemical standards 8.9.1 PFOS (3M Specialty Chemical Division), molecular weight = 538 8.9.2 PFOSA (3M Specialty Chemical Division), molecular weight = 499 04349 8.9.3 PFOSAA (3M Specialty Chemical Division), molecular weight = 585 ExtractionoEfTSF-O84S.1from Serum Page3of 14 } 8.9.4 EtFOSE-OH (3M Specialty Chemical Division), molecular weight = 570 8.9.5 PFOSEA (3M Specialty Chemical Division), molecular weight = 527 i 8.9.6 MSS6 (3M Specialty Chemical Division), molecular weight = 557 | 89.7 Surrogate standard: 4-H, perfluorooctane sulfonic acid (1-H, 1-H, 2-H, 2-H C4F13SO;H) molecular weight = 428 8.9.8 Other fluorochemicals, as appropriate | 8.10 Reagent preparation NOTE: When preparing larger volumes than listed in reagent, standard, or surrogate `preparation, adjust accordingly. ] 8.10.1 11000N0msoLdbieuamkheyrdrcooxnitdaien(inNgaO5H0)0:mWLeiMiglhl-aQpTMproxwiamtaetre,lmyi2x0u0ntgilNaallOHso.lidPsouarreinto a dissolved. Store ina 1 L Nalgene bottle. | 8.10.2 N1 NNsaoOdiHumsohlyutdiroonxiindteo(aN1a0O0H)m:L Dviolluutmeet1r0icNfNlaak OanHd di1:l10u. teMtoevolaum1se0umusLinrogfM1ei0ll | QTM water. Store in a 125 mL Nalgene bottle. | 8.103 0.5 M tetrabuyltylammonium hydrogen sulfate (TBA): Weigh approximately 169 g of TBA into a 1 Lvolumetric containing 500mLMilli-Q" water. Adjustto pH 10 | NusaiOnHg,apapdrdoxsilmoawtleylybe4c4autsoe5t4hemLpoHfch1 angNeNs0 aabOruHpt(lyW)h.ilDeialdutdeintgo tvhoelluamset mwiLthofMilli QTM water. Storeina 1 L Nalgene bottle. . 1 8.10.3.1 TneBeAderdequusiirnegs1aNchNecakOpHrisoorltutoieona.chuse to ensure pH = 10. Adjust as 8.10.4 0.25 M sodium carbonate/sodium bicarbonate buffer (Na:COyNaHCO,): Weigh | `baipcparrobxoinmaatteel(yN2a6H.5COgo)fisnotodiau1mLcavroblounmaetteri(cNfal;aCskOasn)danbdri2n1gt.0o.vgooflsuomdeiwuimth Milli QTM water. Storeina 1 L Nalgene bottle. | 811 Standards preparation . . 8.11.1 Prepare PFOS standards for the standard curve. | 8.11.2 Prepare other fluorochemical standards, fluorochemical standards are acceptable (afsoarpepxraopmrpilaet,e.onMeuwlotrikcionmgposnteanndtard solution containing 1.00 ppm PFOS, 1.02ppm PFOSA, 0.987 ppm PFOSAA, and | 1.10 ppm EtFOSE-OH.) 8.11.3 Weigh approximately 100 mgof PFOS into a 100mLvolumetric flask and record | . the actual weight. 8.11.4 Bring to volume with methanol for a stock standardofapproximately 1000 ppm (ng/mL). ] 8.11.5 Dilute the stock solution with methanolfor & working standard 1 solution of `approximately 50 ppm. ! 8.11.6 Dilute working standard 1 with methanol for a working standard 2 solution of approx. 5.0 ppm. 004350 i ETS841 Page sor1s Extractionof FOS from Serum | 8.11.7 Dilute working standard 1 with methanol for a working standard 3 solution of `approx. 0.50 ppm. 1 812 Surrogate stock standard preparation 8.12.1 Weigh approximately 50-60 mgofsurrogate standard 1-H,1-H, 2-H, 2-H, CF13S0;H into a 50 mL volumetric flask and record the actual weight. 8.12.2 Bprpimn.g to volume with methanol for a surrogate stockof approximately 1000-1200 8.12.3 Prepare a surrogate working standard. Transfer approximately 1 mLofsurrogate stock to a 10 mL volumetric flask and bring to volume with methanol for a `working standard of100 ppm. Record the actual volume transferred. 9.0 SAMPLE HANDLING ] 9.1 All samples are received frozen and must bekept frozen untiltheextraction is performed. 92 Allow samples tothaw to room temperature prior to extraction. | 10.0 QUALITY CONTROL 10.1 Solvent Blanks, Method blanks and matrix blanks | 10.1.1 An aliquot of 1.0 mL methanol is used as a solvent blank. 10.1.2 Extract two1.0mL aliquotsof Milli-QTM water following this procedure and use 1 `as method blanks. ! 10.1.3 Extract two 1.0 mL aliquotsoftheserum following this procedure and use as `matrix blanks. See 11.1.4, 10.2 Matrix spikes - ; 10.2.1 tPheracecuarpnacdayaonfartlhyeezeexmtartarcitxiosnp.ike and matrix spike duplicate samples to determine: 10.22 Prerceepiavreedeiatchhsepaickhe suasimnpglea sseat.mple chosen by the analyst, usuallythe control matrix J 10.2.3 EAxddpietcitoendalcosnpickeenstrmaatyiobnesiwniclllufadlelidnatnhdemmaiyd-fraallnignetohftehleowi-nritainalgecoalfitbhraetiinointicaulrve. | calibration curve. 10.2.4 Prepare one matrixspikeand matrix spike duplicate per 40 samples, with a `minimumof 2 matrix spikes per batch. | 103 Continuing calibration checks 10.3.1 Prepare continuing calibration check samples to ensure the accuracyofthe initial | calibration curve. 10.3.2 Perxeapmaprlee,,aitfaa msiamnpilmeums,eto=ne3c4o,nftoiunruicnhgecchksecakrepperregparroeudpoafnd1e0xtsraamcptleesd. For 10.3.3 PTrroepppeaitrtsealeaccuhrvceo.ntinuing calibration check from the same matrix used to prepare 004351 i! ExtractionoEfTSP-F8O-4S.1from Serum Page Sof 14 10.3.4 Tcuhreve.expAedcdtietdiocnoanlcesnptirkaetsimonasywiblelifnalcllwudietdhitnhatthefamllidi-nrtahnegleoofw-trhaengienoitfitalhcealiinbirtiaatlion calibration curve. This is necessaryifthe analyst must quantitate using only the low endofthe calibration curve (for example, 5 ppb -- 100 ppb, rather than 5 ppb -- 1000 ppb). 11.0 CALIBRATION AND STANDARDIZATION 111 Prepare matrix calibration standards } 11.1.1 Transfer 1mLof serutmo a 15mLcentrifuge tube. 11.12 Ifmost sample volumes are less than 1.0 mi, extract standards with matrix `volumes equal to the sample volumes. Do not extract less than 0.50 mLofmatrix. ] Record each sample volume on the extraction sheet. . 11.1.3 While prepaging a totaloftwentyaliquotsin 15 mL centrifuge tubes, mix or shake i `between aliquots. 11.1.4 Two 1mLaliquots, or other appropriate volume, serve asmatrix blanks. Typically : use the standard concentratiaonnds spiking amounts listedin Table 1, at the end of | this section, to spike,induplicate, twostandardcurves, foar totalof eighteen standards, two matrix blanks, and two method blanks. 11.15 Refer to validation report ETS-8-4,0 & ETS-8-5.0-V-1, which liststheworking. | ranges and the Linear Calibration Range (LCR) for calibration curves. 11.16 Use Attachment D as an aid in calculating the concentrationsof theworking. | standards. See Section 13.0 to calculate actual concentrations of PFOS in calibration standards. 112 To each standard, blank, or continuing check, add appropriate amountof surrogate | `working standard for the concentration to fall within the calibration curve range 5 ppb - 1000 ppb. : | 113 Extract spikedmatrix standards following 12.6-12.16ofthismethod. Use these standards i to establish each initial curve on the mass spectrometer. ExtractionoEfTSP8F4O.S1 from Serum 004352 Page oie 1 Table 1 Approximate spiking amounts for standards and spikes Using 1.0 mL of matrix `Wo(arpkpironxg. stcoanncd.ard uo "Apapnraloytxe. fiinnamlactornci.x of TT Bak | [0s00 |p2p0 m | 0010ppm . I| [s0[0s pp0m 0| |pp 2105 m | 1 0750pm 1.00pm | 2.0 PROCEDURE | 12.1 Owabttearibnatfhr.ozen samples and allow to thaw at room temperatureorin a lukewarm i 12.2 Vortex mix for 15 seconds,then transfer 1.0 mL or other appropriate volume toa 1S mL polypropylene centrifuge tube. y 12.3 Retumunused samples to freezer after extraction amounts have been removed. ; 12.4 Record the initial volume on the extraction worksheet. E 12.5 Label the tube with the study number, sample ID, date and analyst initials. See attached | `worksheet for documenting the remaining steps. | 12.6 sStpainkdeaarldl assamdpelsecsr,ibiendcliundi11n.g2.blanks and standards, ready for extraction with surrogate 12.7 SLinptikheaeatcshemctaitorni,xfwoirtthhethcealiabprpartoiporniactuervaemsotuanndtoarfdss.tanAdlasrodparsepdaersecrmiabterdixinsp11i.k1e,s oarndTable | continuing calibration standards. 12.8 Vortexmixthe standard curve samples, matrix spike samples, and continuing calibration `samples for 15 seconds. | 12.9 Check to ensurethe 0.5 M TBA reagentisat pH 10. Ifnot, adjust accordingly. } 12.10 bTiocaerabcohnastaempblufef,era.dd 1 mL 0.5 M TBA and 2 mLof 0.25M sodium carbonate/sodium 12.11 Using an Oxford Dispenser, add 5 mL methyl-fert-butyl ether. | 12.12 Cap each sample and put on the shaker ata setting of 300 rpm, for 20 minutes. 12.13 Centrifuge for 20 to 25 minutes at a setting of 3500 rpm,oruntil layers are well separated. i 004353 ! ExtractioEnToSf-P8F-O4S.1from Serum Page 7of 14 i 12.14 Labelafresh 15mLcentrifugetubewith thesame informationasin 12.5 . 12.15 Remove 4.0 mL ofthe organic layer to this clean 15 mL centrifuge tube. | 12.16 Phouutrse.ach sample on the analytical nitrogen evaporator until dry, approximately 1 to 2 1 12.17 Add 1.0 mL ofmethanol to each centrifuge tube using a graduated pipette. : 12.18 Vortexmixfor 30 seconds. | 1219 Attach a 0.2 umnylonmeshfiter to a 3ccsyringeandtransferthesampletothissyringe. i Filter into a 1.5 mL glass autovial or low-volume autovial when necessary. 12.20 Label the autovial with the study number, animal number and gender, sample timepoint, ] `matrix, final solvent, extraction date, and analyst(s) performing the extraction. 1221 Cap and store extracts at room temperature or at approximatel4y C until analysis. i 12.22 Completthee extrafion worksheet, attached to this document, and tape in the study i notebook or include in study binder, as appropriate. 13.0 DATA ANALYSIS AND CALCULATIONS ' 13.1 Calculations | 13.1.1 cCaallicburlaattieonacsttuaanldacrodnsceunstirnagtitohnesofoflPloFwOiSng,eoqruaottihoenr: applicable fluorochemical, in I mmLLoofsftasntadnadradrd+xmcLonocfesnutrrraotgiaotneofstsatnadnadradr+d(inuigti/almmlat)r_ ix _ vol_ ume_(m_ L) - = | Final Concentration (ug/mL) of PFOS in matrix 14.0 METHOD PERFORMANCE . | 14.1 The method detection limit (MDL) is analyte and matrix specific. Refer to MDL report C for specific MDL and limitofquantitation (LOQ) values (see Attachments B and C). | 142 tThheeqfuoalliltoywoifntghqeualeixttyraccotnitornolansdamapnalleyssiasre extracted witheachbatchofsamples to evaluste 14.2.1 Method blanks and matrix blanks. | 14.2.2 Matrix spikeand matrix spike duplicatesamplesto determine accuracy and precisionofthe extraction. | + 142.3 Continuing calibration check samples todetermine the continued accuracyofthe initial calibration curve. | 143 Refer to section 14ofETS--5.1 for method performance criteria . 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT i 15.1 Sample waste is disposed in biohazard containers, flammable solvent wastei disposed in high BTU containers, and used glass pipette waste is disposed in broken glass containers ' located in the laboratory. 004354 | ExtractioEnoTfSF8O4S1from Serum Page8ar 1s 16.0 RECORDS | 16.1 nCootmepbloeotkeotrheinecxlturdaecitinotnhweor3-krsihnegesttuatdtyabcihneddert,oatshiaspmpreotphroidat,e.and tape in the study | 17.0 ATTACHMENTS 17.1 Attachment A, Extraction worksheet | 17.2 Attachment B, MDL/LOQ values and summary 17.3 Attachment C, Calibration standard concentration worksheet | 8.0 REFERENCES 1 18.1 The validation report associated iththis method is ETS-8-4.0 & 5.0-V-1. | 182 F'HAPCLTC--ME-le3c.t1r,os"pArnaaylyMsaissosfSSpeecrturmomoertOrty"her Fluid Extracts for Fluorochemicals using 19.0 AFFECTED DocuMENTs 191 ETS-8-5.1, "Analysis of Serum orOtherFluid Extracts for Fluorochemicals using HPLC | Electrospray Mass Spectrometry" 20.0 REVISIONS 1 Revision -- -- Revis-- ion Number `Reason For Revision Date 1 Section 12.21 Changed to include sample storage at room temperature. 04/02/99 | Section 12.13 Addedtheshakerspeed. Section 12,17 Final volume is 1.0 mL; not adjusted for initial volumes less | than 1.0 mL. | ETS84.1 ExtractionofPFOS from Serum 004355 Page alls 3M ENVIRONMENTAL LABORATORY METHOD ANALYSIS OF POTASSIUM PERFLUOROOCTANESULFONATE OR OTHER FLUOROCHEMICALS IN SERUM EXTRACTS USING i HPLC-ELECTROSPRAY/MASS SPECTROMETRY 1 Method Number: ETS-8-5.1 ! Author: Lisa Clemen, Robert Wynne | Approved By: Adoption Date: 03/01/99 Revision Date: | Laboratory Manager Date ) Group Leader Date 1 CE----------------------_ Technical Reviewer Date 1 10 SCOPE AND APPLICATION i , 1.1 uSscionpge:HPTLhCi-selmeectthroodspdreasycr/imbaessstshpeeacntarloymseitsroyf.serumextractsfor fluorochemical surfactants i 1.2 Applicable Compounds: Fluorochemical surfactants or other fluorinated compounds, or other ionizable compounds. 1.3 Matrices: Rabbit, rat, bovine, monkey, and human serum, orotherfluids as designatedin the validation report. 004356 `Word 6/95 AnalysisofSeErTuSm$E-x5tr.a1ct Using ESMS Page 1of9 2.0 SUMMARY OF METHOD 2.1 Tothhiesrmfeitihdso,dudseisncgriHbPesLtCh-eelaencatlyrsoissporfayf/lmuaosrsocshpeemcitcraolmesturryf,acotranstismielxatrrsacytsetdemfraosmapsperrouprmioarte. `The analysis is performed by monitoring a single ion characteristic ofa particular fAdlduiotrioocnhaelmliyc,als,amspulcehsamsatyhebpeearnfaluloyrzoeodcutsainnesgualtfaonnadteem(PmFasOsS)spaencitonr,omme/tzer= the identity ofa compound by detecting daughter ions ofthe parent ion. t4o99f.urther verify | 3.0 DEFINITIONS ] 3.1 syAsttemmossaplhleorwifcorPrvearsisouursemIeotnhizoadtsioofnio(nAiPzTa)t:ioTnhebyMiutcirliozmiangssvaQruiaotutsrsoouIrIctersi,plperqoubaedsr,uapnodle Pirnteesrsfuarceesc.heTmhiecsaeliInocnliuzdaetibount(aArPeGn)o,t Tlihmeirtmeodstpo:raEyl,ecettrc.ospTrhaey iIoonniizzaattiioonnp(rEoScTe)s,siAnttmhoesspeh.eric 1 techniques occurs at atmospheric pressure (i.e. not under a vacuum). 32 presEsulreec,trwohseprreabyyIioonniszaintisoonlu(tEiSon, aErSeI)t:raansmfeertrheoddtoofithoenigzaastpihoanspeevrfiotrimneydcahtaragtemdosdprhoeplreitcs. | These charged droplets are produced by the application ofa strong electrical field 1 33 (MSM/aMsSs):Sp`eTchterAomPeItQruya,tMtraosIsI tSrpieplcetrquoamdertueprol(eMSs)y,stTeamsndareemeqMuaispspeSdpweictthrqoumaedtreurpole mass sseulbescetqiuveendteltyecdteotresc.tedI.onsAarseinsgelleeMctSivemlayydibsecreimmpinlaotyeeddbfyomraisosn dteotcehcatirogneorrataiose(rmi/ezs)(aMnSd/MS) 1 for more specific fragmentation information . : 3.4 triplCeoqnuvaednrtuipoonlaelsvyss.teZm-ssp(proasytp1r9o9b8e) iuntitleirzfeaace":Z-Tshprealya"tecsotnmfoordmealtsioonf.MiTchreomsapsrasyQeumaittttreodIT fdriormecatapltryotbheeicsoonrethaopgeorntaurlet,otafhteercopansesianpgertthurreo.ughInattohretcuoonuvsenptaitonhawlaycoinnftohremactoiuonnteirt is aimed electrode. Though the configuration is different, the methodsofoperation, cleaning, and i `nmoaticnotmepnaantcieblaerewitthhe osanmee.anoHtohewre,veburt,oZn-lsyprwaiythcosimmpiolanrensytsstaemnsd (cio.n.v,eZn-tsipornaaly ccoommppoonneennttss aarree compatible with some other Z-spray systems, etc.) ] 35 QuatMtarsoIs LtyrinpxleSqoufatdwraurpeo:leSsyysstteemms.sofCtuwrarreentdleysMigansesdLfyonrxthheasspWeciinfdicoowpser9a5tiaonnod fWtihnedsoe.wsNT | i4n.s0tvreurmseinonts.(MiAcllrovmearssisoQnsuaatrtersoimIiTlatrr.iplFeoqruamdorruepodleetaMilasssseLeytnhxe moranMuaaslssLpyencixfiNcTtoUtsheer's Guide). 4.0 WARNINGS AND CAUTIONS | 4.1 Health and Safety Warnings: -- ' 4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe |1 employs a voltageof approximately 5000 Volts. 004357 || ETSS51 Page20r9 `Analysis of Serum Extract Using ES/MS | 4.1.2 Wanhdecnlohtahnidnlging samples orsolventswear appropriate protective gloves, eyewear, 4.2 Cautions: 4.2.1 DIfothneotbaocpkerparteesssuorleveenxtcepeumdpss4a0b0obvare, ctahpeaPci1ty1o0f400w0ilbainrit(i5at8e00auptsoim)abtaicckpsrheustsduorwen.. 42.2 Do not run solvent pumps to dryness. i 5.0 INTERFERENCES 1 5.1 Tstoormaigneiomrizaenyinptaerrtfoerfeinncesstwrhuemneanntaalytztiihonagtnscaommpleess,intceofnlotnacsthowuiltdh ntohtesbaempusleedorforexstaramcptl.e 6.0 EQUIPMENT ~ ] 6.1 E`mqoudiifpimceantitonlsisitnedtbheelraoww m ataba aesmmoedity fhioedddienvioartdieonrst.o optimize the system. Document any | 6.1.1 MelieccrtormosapsrsayQiuoantitzraotIilotnrsiopulrecqeuadrupole Mass Spectrometer equippedwithan 6.1.2 HP1100 low pulse solvent pumping system, solvent degasser, column compartment, | and autosampler , 7.0 SUPPLIES ANDMATERIALS . | 7.1 Supplies 7.1.1 High purity grade nitrogengasregulated to approximately 100 psi (House air 7.12 syHsPtLeCm)analytical column, specifics to be determined bytheanalya.st and documented inthe raw data. | 7.1.3 Capped autovials or capped 15 mL centrifuge tubes 8.0 REAGENTS AND STANDARDS ] 81 Reagents 8.1.1 Methanol, HPLC grade or equivalent I 8.1.2 eMqiulilvail-eQnTMt,waatnedr,maalylwbaeteprrouvsieddeidnbtyhiasMmieltlh-oQdTsOhoCulPdlubsesMyisltleim-oQrTMowtahteerrvoerador | 8.1.3 Ammonium acetate, reagent grade or equivalent 82 Standards 8.2.1 Typically prepared dtuwroinmgetthheoedxtbrlaacntkiso,ntpwroomceadturriex.bSlaeneksE,TaSn-d8-e4i.g1h.teen matrix standards are ! 9.0 SAMPLE HANDLING | ETS 851 AnalysisofSerum Extract Using ESS 358 Page3ary 9.1 aFrreesshtomraetdriixn csatapnpdeadrdasutaorveiaplrseoprarceadpwpietdh1e5acmhLancaelnytsrisi.fugEexttruabcetseudntsitlanandaalrydssisa.nd samples 9.2 Ifanalysis will be delayed, extracted standards and samples can be refrigerated at approximately 4 C, or at room temperature, until analysis can be performed. 10.0 QUALITY ConTROL 10.1 Solvent Blanks, Method Blanks and Matrix Blanks 10.1.1 aSoclhvebnattbclhantkosd,emteetrmhiondebcloanntkasmiannadtimaotnroirx cbalrarnykosvearr.e prepared and analyzed with 10.1.2 Analyze a method blank and a matrix blank prior to each calibration curve. ] 10.2 Matrix Spikes . 10.2.1 Matrixspikesare prepared and analyzed to determine the matrix effect on the 1 recovery efficiency. | 10.2.2 Mraetcorviexrsypfiokre edaucphliacnaatleystea.re prepared and analyzed to measure the precision and the 102.3 Analyzae matrix spike and matrix spike duplicate per forty samples, with a `minimum of2 spikes per batch. 10.2.4 Matrix spikeand matrix spike duplicate concentrations will fall in the mid-range of | rthaengienoitfitalhcealiinbirtaiatliocnalciubrrvaet.ionAdcduirtvei.onal spike concentrations may fall in the low- . 10.3 Continuing Calibration Verifications 10.3.1 Continuing calibration verifications are analyzed toverifythe continued accuracy of the calibration curve. " | 103.2 SAnpaleyzeeaaimiad-range calibration standard ; aftreverytenth sample,with a minimum ] 11.0 CALIBRATION AND STANDARDIZATION 11.1 Aanvaelryazgeetohfetewxotrsatcatneddamradcturrivxestsawnidlalrbdespprliootrttedobaynldifnoealrlroewgirnegsesaiocnhs(eyt=ofmexy+trabc),tsw.eiTghheted | 1x, not forced through zero, using MassLynx orothersuitable software. 11:2 Ifthe curve does not meet requirements, perform routine maintenanceorreextract the | "standard curve(ifnecessary) and reanalyze. 11.3 For purposesofaccuracy when quantitating low levelsofanalyte, it may be necessary to I i1 uEcsaxelatimbhprealteil:oonwwechunredvnoeafcttotnhesemipcstatliiinnbggortafottiqohuneacnusttriavtnaedtraearatdphspefrrortxohimamn5attpheeplfbyutl1ol0r1pa0pn0gbepoopffbantrahaletyhtseetr,atnghdeaannredrtcahuterefvuaell. rangeof the curve (5 ppb to 1000 ppb). This will reduce inaccuracy attributed to linear f regression weighting ofhigh concentration standards. 004359 AnalysisofSerEuTmSEx8t5ra1ct Using ESAS Page dors 12.0 PROCEDURES 12.1 Acquisition Set up 12.1.1 fClliecnkamoen ustsairntgbMuOtt-oDnAiYn-tlhaesAtcqduiigsitiotifoyneCaorn-tsraomlplPeanneul.mbSeert,uapssaigsnamapmleetlihsto,d A(sMsSi)gna for acquiring, and type in sample descriptions. | 12.1.2 TSIoRcr(eSaitneglae mIeonthRoedcocrldicikngo)nosrcaMnRbMu.ttonSienttIhoeniAzactqiuoinsiMtoiodnecoanstarpoplrpoapnreilataendansdelmeacsts. 1S0IR4s9.9Soarvoethaecrquaipspirtoipornimaettehmoads.seIsf. MAS/fMulSl sicnasntrisumuesnutasllayrceoellmepcltoeydeadl,oandgdwitiitohnatlhe 1 `product ion fragmentation information may be collected. See Micromass MassLynx `ReGaUcItDiEonTMOotDoAriTnAg)A.CQUISITION for additional information and MRM (Multiple ] 12.1.3 Typically the analytical batch run sequence begins withasetofextracted matrix standards andendswith a setofextracted matrix standards. | I 12.1.4 Samples sample. Saroelvaennatlbylzaendkwsisthhoaulcdonbteinaunianlgyzceadlipberraitoiodniccahlleycktoinmjoencittedoarftpoesrseivbelreyatneanlytthe carryoverandarenot considered samples butmaybe included as such. | 12.2 Using the Autosampler 12.2.1 Set up sample tray according to the sample list prepared in Section 12.1.1. 1 1222 aSneatl-yuspttchoensHiPd1e1r0s0a/papurtoporsiaamtpelfeorraotptthiemaflolrleoswpoinnsge.conRdeictoirondsaocrtuaatlccoonnddiittiioonnssitnhethe | instrument logbook: . 12.2.2.1 Sample size = 10pL injection - | 122.22 Inject/sample = 1 12.223 Cycle time = 13.5 minutes ] 12.2.2.4 Solventramp = Time 20mM | | | Ammonium acetate [00min | "a0% | 60% | [Bs0min [00% [10%| [Lomin [00% [10% | 020min| a0% [60%| | 12.225 Press the "Start" button. i 123 Instrument Set-up i 12.3.1 Refer to ETS-9-24.0 for more details. ! 123.2 Check the solvent level in reservoirs and refillif necessary. 004360 | AnalysisofSeErTuSmSt-r5ac1t Using ESMS Page Sof9 ! 12.3.3 Check the tip. thTehsetatiinplseshsousltedelbecafplialtlwairythatntohjeaegngdedofetdhgees.proIbfet.heUtsipe iasnfoeuyenpditeocebet.o check unsatisfactory, disassemble the probe and replace the stainless steel capillary. 12.3.4 OSbetseHrPveLCdrpopulmetpstcoo"mOirn"g. oSuettofthtehefliopw tooft1h0e-pSro0b0e.uLAl/lomowritaonseqaupiplriobprraitaetef.or approximately 10 minutes. | 12.3.5 Taruorunnodntthheetinpiotfrtogheen.prAobef.ineRmeiasdtjusshtotuhledtbiepeoxftpehlelepdrwoibtehifnnoonimtirsotgeisnloebaskeirvnegd. | 12.3.6 cThhaenignestinruomrendteutorseosptthiemsiezepatrhaemreetseprosnastet:he following settings. These settings may ] 12.3.6.1 Drying gas 250-400liters/hour 12.3.6.2 ESInebulizing gas 10-15 liters/hour 12.3.6.3 HPLC constant flow mode, flow rate 10 -- 500 uL/min 1 12.3.6.4 HPrPeLssCuries <op4e0r0atbianrg(cTohrirsecptlayr.)ametiesr not set, it is a guide to ensure the | 12.3.7 fCuarrtehfeur.llCygounindeectthteheprvoolbteaignetocatbhleesopteontihneg.proIbnes.ert probe until it will not go any 1 12.3.8 tPraipnetdtihnettoutnheepiangset,ruwmietnhttlosg.parameters, and stoirt in the study binder with acopy | 12.3.9 aUnsailnygsitsohefcbriooslso-gfilcoawl counter electrode matrices in the ES/MS source is recommendedforthe. 1 12.3.10CMlaiscksLonynstvaerrsbiuotntso,nsienetahpeprAocpqruiiastietiMoansCsoLntyrnoxlUPaSnEelR(SthiGsUmIDaEy)v.aryPraemsosntghe start i button. Ensure start and end sample number includes all samples to be analyzed. | 13.0 DATA ANALYSIS AND CALCULATIONS 13.1 Calculations: 1 13.1.4 Calculate matrix spike percent recoveries using the following equation: | % Recovery = OBbsearvecdReksulgt- rounResdult x 100 Expected Result 13.15 Calculate percent difference using the following equation: | % Difference = Exp CoEnxce p.e-cCtc aeldcCut olnact,e edCod nc, x 100 ' 13.1.6 Calculate actual concentrationofPFOS, or other fluorochemical, in matrix (ug/mL): i foros _---- . (itilVolumeofmattix (mL)+mLofSumrogateStandard) 1000 ng Final Volume ha(mL) 004361 ETSS.51 AnalysisofSerum Extract Using ESMS Page6ars | | 14.0 METHOD PERFORMANCE. . 14.1 M`meattrhioxdspDeceitfeicc.tioPnleLaismeitse(eMEDTLS)-8a-n4d.1L,imAitttoafchQumaennttitBa,tifoonr(LlOisQt)ingaroefcmuertrheondt,vaanlaildyattee,dand ] MDL and LOQ values. 142 Solvent Blanks, Method Blanks, and Matrix Blanks | 14.2.1 lSoowlevsetntstblaanndkasr,dminetthheodcablliabnrkast,ioanncdurmvaterix blanks values aremustbe below the | 143 Calibration Curves 14.3.1 The r* value for the calibration curve must be 0.980 or better. 1 14.4 Matrix Spikes | Somtoing | 14.4.1 cMoantcreinxtrsaptiikoen.percent recoveries are must be within + 30%ofthe spiked ! 145 Continuing Calibration Verifications 14.5.1 Continuing calibration verification percent recoveries must be 30%ofthe spiked ] 14.6 Ipfecrrfitoerrmieadliosntetdihnetshyisstemmetahnoddspaemprlfeosrmraenacnealsyezcetdioonriosnt'htermeatc,timoanisnatsendaentceermmianyedbbey the analyst. Document al actions in the appropriate logbook. 1 14.7 Iffodoattnaotaerdeotno btaeblreespoarntdeddiwshceusnsepderifnotrhematnecxet corfitthereiarehpaovretnot been met, the data must be : | 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT - | 15.1 Sample pipette extract waste s wdaisstpeosaenddifnlbarmomkaebnleglsaoslsvceonnttiasidneirssplooscaeitdnedhiighthBeTlUabocroanttoariy.ners, and glass 16.0 Recorps | 16.1 Eheaacdherpaogrehgaennderwartietdenfoornatshteudpyagmeu:ststhuadvyeotrheprfoojlelcotwniunmgbienrf,oramcaqtuiiosnitiinocnlmuedtedhoedi,thenr the | ainnatleygsrta.tion method, sample name, extraction date, dilution factor(iapplicable), and 16.2 Parpipnrtoptrhieattuensetupadgyef,olsdearm.plCeolpsty, athnedsaecpqaugiseistiaonndmteatpheoidntfortohmeMinasstsruLmyennttrounilnocgl,udeinthe 16:3 Pstloortetihnetchaelisbtruatdiyonfoclduerrv.e by linear regression, weighted 1/x, then print these graphs and 164 Parnidntstdoarteaiinnttehgersattiuodnysfoulmdemra.ry, integration method, and chromatograms, from MassLyn, 16.5 ASutmmtaraiczehdAmatfaeounrsaitnng esuxiatmapblleesooffatwsauremm(aErxcyeslp5r.e0a)dsahnedets.tore in the study folder, see 004362 AnalysisofSeErTumSESxt5ra1ct sing ES/MS Page 7019 | 16.6 Banadckloucpateiloenoctfrobnaicckduaptaetloecatprpornoicprdiaattae medium. Record in study notebook thefilename 1 17.0 TABLES, DIAGRAMS, FLOWCHARTS, AND VALIDATION DATA. I 17.1 Attachment A: ETS-8-5.1 Data summary spreadsheet 18.0 REFERENCES | 18.1 `FcAoCmTp-oMu-n4d.s1f,fo"EmxStrearcutmiofnoorfPAnoatlayssissiuUsmiPnegrfHlPuLoCr-oEolcteacntersouslpfroanayt/eMaosrsOtShpeerctFrloumoertorcyhemical ] 18.2 EToTnSi-z9a-t2i4o.n0/,Ma"sOspeSrpaetcitornomaentderMaQiunattetnraonIceotrfiptlheequMaidcrruopmoalsesSAytsmteomssp"heric Pressure 18.3 The validation report associated with this method is ETS-8-4.0 & 5.0-V-1. 19.0 AFFECTED DOCUMENTS | 19.1 ECToS-m8p-o4u.n1,ds"EfxrtormacSteiornumoffPoortAansasliyusmisPUesriflnugoHroPoLcCt-aEnleescultfroonsaptreaoyr/OMtahsesrSFpleucotrroocmheetmriyc"al | 20.0 REVISIONS Revision Revision | SSeeccttiioonn plotting 61l1.i.11ne.Aa2r rCvelgarrieefsiscoiaroftnitoaanwnoodcfgauHrdPvd1eee1sd,0tn0hoetsy1s/stxtaenwmediacgrohdmtvpiaonlngueenosft,tsah.reecuusreved.fo r 04/02/99 | SSeeccttiioonn 1172..12.C2.h4aCnlgaerdifficraotmioantotfsaoclhvmeentnBtratmopA.. . 1 , . AnalysisofSerEuTmSEx8t5ra1ct Using ESMS 004363 Pages ar Laboratory Stud#y lw | MTeastsMxaFtinelrSlvat: . MetbodRevision: Analytical Equipment System Number: | RFIinelsSetqnrauummieee:ndt Softwa Value: re/ Vers ion : 1 SlDYoiupneeto:efsEexpv:acton/Aadlyst: 1 Date of Analysis/Analyst: EE ra aEl a | SGirpaeu:p/TDatkee:nTfarkoemnifnreosm rheepsesuionfoeldnert.i SCoanmcpelnet:ratTiaokne(ngf/ommL)t:hTsatkudeynfrooldmerheMas yx gration summary. | Intal Volume(mL: Takenfromth study folder i Dilution Factor: Taken from the study folder. : Final Cone. (ug/ml): Calculated by dividing the initial volume from the concentration I | ! AttachmenAt:SummarySpreadsheet ETS-8-5.1 Analysis of Serum Extract Using ESMS 004364 Page 9of 9 | BEST COPY AVAILABLE|.Fl.uo.rine Analytiical Chemisi try Team \ 0Coe [rm . | QuDaenttietraBtriount:ion of PFOS LmterDemimaon in sera by Electrospray Lope MaSpsescifSipcietcy:trometry RettonTime: (ESMS) 73min. . LiRmaingteooffQuCaanltiibtraattiioonn(CLur0vQe):: LL0opppeb- 100pp Moleolarloe: 459emu | Commision Contin @ 09 | PHASEIAIv:erAsngaelcyosnies.ofpoSoilded se% Ur ReScs o1v9e, r9y8 Let PEOSGoW De MS MSD Tvnpone Sd Le EEOSGob' Dex Rewer MS MSD T0o0s7 "44 62 Jaermae wwuose 4oa 38 sw e m@ | Sn0s9 2436 28 eun we2 wre 3 as ose si 7 ve me ] swooiitz 4sse 132 1E00 E)108 | wwoou a50 56 005 1920 [RanogfAeverags ConcenoftPFrOSa1tLiotSoerna: __fpprb1o:5m4p6pb | Resultsof QuantitationofCoMnecetnhtroadtBiolnanks Blank REOS(ooh) | {MethodBleak, F20-1 <LoD ethod Blank, H20-2 <LoD [MethodBlank, sera-1 (1) | MMeettihooddBBllaannkk,,sseerraa--11 G(32)) | [MethodBlank, sera-2 (1) 7 <<opp <LoD Sample' Qc Qe Qs Qs [ros] recovered 49 49 4s9t 53 [FOS % expected recovery 49 100 49 100 449 100 49 108 SReypsrtoedmucRlepyr'osducibility and Precision Checks ] Sioa0msp:3le2) EEO4Ss@pb oRSmD | w50o0z9--33(22)) | wwoorisr:3s G2y) : 1514 o0u4s% 33 o1s8% | = S10a0m7-p1l(e1) T1000700-771-11 3(26))) 1007-1 5) Te Std.Dev. RSD RES"(7ob) 5o5 1 2% | 0"RCeposratmepdlevsalauees eaxrteratchtesaovferbagaesnscerkosnpickeed wniPtthFPOrRSaOoSftsaimdpoalenrsdextractiend triplicate. Reproducibility samples are repeat injectionsof lot sera extracts, perfoafrtermeveerdy 15 samples. "Precision samples are 5 consecutive injectionsof the same lotseraextract. 1 3E*MQouEenstviiornosnbmleenDtaaltaL.abSource oferrorcurrently undetermined. | 004365 PHASED ] BESTUUPY AVAILABLE :; rruorine Analytical Chemistry Team RiConse: .Je.cemid _onK in sera by Electrospray Mass Spectrometry (ESMS) | QLuiamnittiatarDfeht.ton omy: 105 Re`ScpeociifiocintTy:ime: Limitof Quantitation (LOQ) : 1.0ppb Molecular fon: `RangeofCalibration Curve: 10ppb - 100ppb 1 CurveCorrelationCoefficient(): 0991 60min, 413 amu' 369 amu? 86018] | PhaseII:AvAenrapeicsonocfpooSlIeLd sera% (POAA) Recovery Kversgroone Sid % Recovery 1L0o6t POA<AoGpoDY De0v. Ms*S MS%D LWeoIts POA<ALGoDON' Deov M0s0 M1s01p | 10078 <<Loopp 1 s%e 79 Wwoolrs <<poD o0 5Bo9 soo <op 1 e+ + BOI <LOD 88 | sS0o10 <<wLoopD 0 47s e57r0 BBosz aODD 00 01080 11000 wor <op 1% mW BOA COD 0 oro 1 wwoole3 <<LwoopD 0 0ER0 100 BOOS <OD 0 9% 2 [Range ofAverage ConcenotfPrOAaItaLiotSoerna:_ allvaueswerebelothwe LOD/LOQ (10950 [-- `Quantitation ofMethod Blanks 1 MBeltahsokd Blank,F20-1 `Concentration BO<AoApooh) | p[Metthod Bslank, eH2m0-i2 r <<oLopD [Method Blank,sera-1 @) <wop [MethodBlank sera-1 3) <op | [eotBank may <LoD Sumole', caL2 oc BA Qcs [reFopOvAerMcd 994 01 2 9 PexOmAecAteld reso%ven 9o%s o9s8 os %. 9o%s 97 97 SystemReproducibility andPrecisionChecks | [eric S0a0m6.p3l2e) POATAoGrea oESmD | fsa 300 70% W012-3 (2) 6500 0.0% | pw[Wooor1irs-s-33o@(n2)) 4wa70o00 as0.e0on% | recon" S1a00m7p-l10e) 1007-12) 1007-13) Pea3k8A0r0ea 3600 6300 T1000077-11 5(4)) Tl66330000 RSSwDope. 3%00 "lon m/s=413used forpurposesof quantitation. | "Reopnormt/es=3v6u9eussedrfeortphuecrpeorsaegsoefscveornfciecnattiroant.ioofncPoOmApoAounfdsaidmepnltietsy.exacteidn pice "QCsamplesareextractsof blankseraspikedwith POAAstandards. | Reproduciblesamplesare reptinjectofibootsnersaestrcts, perfofrermeevedry 15 samples. 66 | P*rQeuceisstioonnasblaempDlaetsa.arSeo5urccoenosfeceurtriovreciurnrjcaetclytuoinfodtenthseersmianmeedl.otsera extract. - 0043! ! ssMEnovironmentat Lab | PHASE 11 POAA) KJ. Hansen Phase INT SMEavirnneaallsd Ouanitative Analysis of PFOS in Individual's Sera DeterminationofPFOS in sera by Electrospray Mass Spectrometry (ESMS) | LL`ioQnutainottriotfeaQtsuiasonntii:otantiogno()LO:G): RangeofCalibration Curve: 1L00pmh M`SoRpelecoimfoiilcirntylT:omwe 74339maimsn Lop oop | Curve Correlation Coefficient (7): ~~ 1 Results of Quantitation of Individual Sera Samples | LotB`PAs vReOraSg@emcoonbc. %R"aSD Lemts PvEerOa4Sg6e@cuonbc. %R15SD 1 m ma 4 "2 os 2 Na mm8 smw 130 ws w 2 | 5o m 98 s a67 N70A m4 350 21rn9a8sw48 B91 use 3s 2 | o so s3 ns ume n 2w8 sa m1e ne 5 15 | 1Lu29s8ma347s N1a 2 2W 8898 ge3s N7A se ] m88 % 21 Fr 1 | onNlAyenNouoghsaamplceflori1nantalayssisews - 3m A - . 004367 Quantitative Analysis of PFOS and POAA in Individual's Sera 411195 DQueatnetirtmaitnioant:ion of PFOS/POAinA serabyElect`Srpoesctpircatyy:Mass Spectrometry(ESMS) ! !|| `LoifDemteictiton (LOD) LCRioammmiotgloeifooQnfuaCCntaioltabdtrioinnCLueOrv:Qe): : 1.0 ppb/10 ppb `Retention Time: 10.L505p90pb/-/011p9009p09ppppbb./b10 pp`Mbo-l1e0c0uplparbloa: 7.35 min/6.9 min. 499emweP/mru,43e61l9 omi3mminary | ResultsofTQuapntitatSioLn of IndAivpideuasl SeraSSiamp Df PEOSGubY Dev 'POAAGEY Dev) If Tropes PROSGobY Bi Den Avene SE POAA(MmM De i 129%8 43%5 oadd <4400Q ods nd| 1798 4s1 adnd <<00QQ amdd i 398 45 ad 18 3 1998 mn ad <Q nd' 4S98 8732 madd <100Q nda ad| 2098 445 p1e <<00QQ andd | 698 53 4 <0Q nd| 2298 54 ad <0Q ad 798 42 ad .. 4oQ nd| 2398 n ad 12 nd 898 49 17 a0Q nd| 2498 28 ad <0Q nd | 9w9s8 3s7m aadd <d00QQ aadd|e25m98 34 0 1ad <400Q Q andd | 1n19s8 | 339 aadd <<00QQ andd| z27e9e8 501 nndd <<0QQ andd 1398 45 ad <0Q ad| 2998 41 nd <Q ad ] 11549988 2%86 ad3 <010Q n1d|| 33098 4b1] aadd <<QQ andd 1698 kl ad <Q nd | Resultsof Quantitationof Method Blanks | B[Mletahokd Black120-1 [REOLSOIDook) [MethodBlank, sera-1 (1) 4 | [Method Blank,sera-1 (2) <LOD bod Bunk120-1 0D [ROAAL(ooh) ] [Method Blank, sers-1 (1) [Method Blank sers-1 (2) <LOD _<LOD Samp [rsFeOovSar]ed Ql 99% Qc 9% Qs 0% _ -rePsOovAeAr}ed 93% 7% 2% | Ewa: tem Reproducibility Checks Sample PROS RSD || 0i10099.5188-:222.(G)2)) F3Y 431386%% 698-1 (2) a 3.0% roAAGra) RSD 21W850o00 000.000%%% 200 61% i"nRehpaorvtedavavlsue3saarelfrromoafsisnagmlpelaenwaalryseiswnleosnsdaansdtd.vdeevr.giseindicated; \ QC samples are extracts ofblank sera spiked with PFOS/POAAstandards. : Reproducibility samplesarerepeatinjectionsoflotseraextracts,performed ev15esarmplyes. 004368 1 | Xi Bama dl Phase II MEmvirouneaal 45 (Quantitative Analysis of PFOSinIndividual's Sera | Determinationof PFOSinserabyElectrospray Mass Spectrometry (ESMS) `Quantitation: | LimitofDefection(LOD) : Lop ReStpeecnitfiicointyT:ime: ~~7.35min. LimitofQuantitation (LOQ) : 1.0ppb Molecular lon: 499amu `RangeofCalibrationCurve: 1.0ppb - 100ppb | CurveCorrelationCoefficient (7): 1 Averageconc. ResultsofQuantoifItnda ivitduialo Sen ra Samples ro | Le PROoSmGoM %EaSD LTeo PRO4S6Gon %31E5SD | m ma 4 u2 urmo 56 130 osm m6 NoA, n umw. 25 | om 7 om @ 5 m4 798 "2 aNA | 0980s39 n5 us 6 u 1 nw 3 1 nm 4s NA um 2398 73s 1 w29s8 451 21 Er 15 8% 3 NA 298 6s 7 | 11o 489588 n391% 1212 3a098 637 01% NA. =No RSD calculatedastherewas . | only caoughsamplefor 1 analysis. mrss : . | . 004369 | FluorineAnalytical Chemistry Team | Determinationof Quantitation: PFOS in seraby Electrospray Mass Spectrometry Specificity: (ESMS) | LimLiimtiotfaQruDaenstciitoatnicono(nL)O:Q) : 10ppb RetestionTime: 10p Molecularlon: 7.3Smin 499 amu RangeofCalibration Curve: 1.0ppb - 100pp | Curve Correlation Coefficient (7): ~~ 1 PHASEIII: Analysis ofindivisedraual samples, 1998 US "Average conc. "Average conc. | Let13P8EOsS(ob) %aReSD LmetsEFOS4(6 ob) % 1R5SD a 1 18% 36 10 1 | mos 52 ow on ss 6 NA, [I 008 40 2m 4 2 9 | & 5 3 moe "a NA |} 88. 49 35 98 39 1 1098 54 ns 6 5 n ] ns 3 1 BB 4s uss 31 NA n ] 15% 68 79% 21 28 72 1 8 38 2 298 a1 330 85 1 298 57 15 28 3 NA 23909988 66s7 s7o 33 1 | NA =o ASD culatedas there was - `only enoughsamplefor 1 analysis. o 6018 . . . | | I smite srs 004370 PHASET | 3M Environmental Laboratory- Fluorine Analytical Chemistry Team KrisHansen -Sr.AnalytCihecmaislt ] BFuliulodriinngeAn2a3lyEt0ic9al Chemistry Team B 127786018 | Phase IV: 'PFOS analysis of US human sera samples collected in the 1970s i-- . `Twelve frozenserasamples of various volumesweresupplied to theEnvironmentalLabbyJeff | us`inMg aan(i3onnM-Mpdeadirieicnagllr)e.aOgennetm. TLhoerexntoramcotrsewtehraenanoanleyhzaeldfqoufatnhteitvatoilvuemlyefoofrePacFhOsSabmyplHePwLaCs-eExStrMaSc.ted Analyteidentificationwasverifiedbycomparisonofmolecularion-HPLCretentiontimeofthe extracted `analyteandstandardmaterial.Sampleswerequantitativelyevaluatedagainst a specially prepared | `extracted curve. `The PFO"STlheevtewleslwveerseasmipglniefsiccoannttlayinloewdaesntahvaenriangtechoenc3e1nstarmaptlioens ocofll2e7cptpebdfPrFomOiSnrdainvgiidnugaflsrionm022-/9488apnpdb. `sbusdanceofthebranchedchainisomerwas37.3+/- 4.8%compared t 3.7+.-7% inthe1998samples. | IdentsatialneddsarudmsmolaurtyioonfstohfePrFeOuSl,stihseibnrcalnudcehdedchainaccountsfor 217+/-3.1%ofthetotalconcentration. A `TheidentityofPFOSinseveralrandomlyselectedsamplesinthis phasewasconfirmedusing 'HPLC-ESMSMS.Inallcases,theanalyteexhibited acharacteristicretentiontime, acharacteristic ] primaryion,andfivecharacteristicsecondaryions. | Experimental summary: 5 Samplepreparation: lon-pairingextraction - In a pH controlledenvironment, an jon-pairingreagent,tetrabutylammonium(TBA),is usedto | `extractthe analytefromthematrix.Thecationicreagentselectivelytargetsanioniccompounds,likePFOS and POAA.SubsequenttotheformationoftheTBA-anionpair,theanalyteistransferredto anon-polar `organicsolvent (ethylacetate),dried,andreconstitutedinmethanolforMSanalysis. ] HPLC:CharacrettenteiorntiimessfotrPiFOcS In HPLC,analiquotoftheextract sinjectedandpassedthrough achromatographiccolumn. ] `pBhaaasmseepdpolan,sitsihnneagastftfhairnnoidtuaygrhodtfshtoelhuectaoinloaunlm,yPnt,FefOtohSretmahnaeaysltyacttleiuosenraaetrtya9-i.pn1hmeaidsnfeuoitnetaschh.eacrRoacltueemrnisrtelitlcaiatmmieaovsebutenottttwohefeeltiniiqmaueios.dtaFmnoodnrbairlde- | + [HPPFLOSCssoylusttieomn.andtheanalyteextracted fromserainthisanalysiswerematchedtowithin1%onthe ESAS: Detectionandmonitoringofthe molecular lon | Analysis ofPFOSstandardsindicatesthattheprimaryion characteristicofPFOSism/z =499 1 `amu, correspondingtothemassoftheanionicsurfactant(CyF;SOy-). `Thisionwasmonitoredselectively tomaximizesensitivity. Ascanofm/z=100to 1210 (negativeonly)wasalso collected. HPLCESE-SM-SM/SMMSS:isDveerystimeialancrdttmooEniiStMooSri,nnegxoefpcthtahraacttietridsdtsicasencaodndidtairoynailodnismension ofcertainty to `compoundidentification. AsinESMS, a charactioenrisisseltecitecd. Afer secttheiESo-MnSM,S 004371 Eraigdertectmedaet thadettectsor. | characterizestheion furtherby smashingitapartwithhighenergygas. As result ofthesmashing, i `secondaryionicfragments (daughterions),characteristicofthemolecule, arecreatedand detected. 1 Forexamplefo,r PFOSanalysis,ion 499isselectedasthecharacteristicprimaryion.Thisionis `smashedinto otherions suchas 80 amu (correspondingto SO), 99 amu (corresptooFSnOdy)i,1n3g0 i amu(correspondingtoCF3S0y), 180amu (CFS05),and230amu (CsFeSOy). Eachofthesesecondary 1| Q`umaatlriitxysDcpioukneetsrtcooolsusalmudpmplmoealbrieymei:tvaatiiounast,sdeorrtshaemspalmepslweesr.eInnsotteeaxdt,rtawctoesdaimnpdluepsliocfaptoeo.lFeodrsteriaspsuarmceharseesdofnr,om '| cSaicghmsaawmeprleeutsoedcfnuorrsducploimcapteematriaxtsiopnikoefatnhaelsyasmisp.lMee.thAamniodleblvalnkqusawleirtyeasnoalmyezedbepflorweaasnsdma after. ] `abQfrutaaecnrktteihtteiasntiigcxountorhvsfeetsrh.eassSpaaemmcppillfeeis.c`QiTChbepafasoerudarompenotitenhrtselaxitenreaaacvtireldeaecbuslrsevoieonwnaetsqhueaaanptapileoynnzddeeedrdbirevefseoudrlfetoasnmtdabtalhfete,earvtehreagseaompfltehse.two | poten 1 HTaPsLtrCusmyesnttealmspecifics: HemetPackardSeries 1100LiquidChromatograph 1 Column:KeystoneB2exta1s0il0Cmyms ym particlesize | FlSSoooilwvverenanttteA::: 2M3.e00t0hmaLMn/aAmlimnm.onium Acetate 1 Salven Gr4adi5ent:%1Bi0n9.950mi0ns. RFeotludrantto904%5588fiorn31..550m0miinnss.. Holdat 45%Bfor 3. mins. Injoc/tsaimoplne:s1 - R .: | fren Electrospraymassspectrometer . | Micromass Platform IIAPIMassSpectrometer " Mass2.L1 Syofn twaxre ] SCoMouondreec:veotllteeamcgtpereso:rsa-pt2rea0y:vn,eg6a0tviv1e15C. . Analyzervacuumpress0.0u00r04e3msB:,0.000079mBar Electrode: cros low 004372 i Fluorine Analytical id Chemistry Team 778-6018 SSaammpplleess RExetcreaicvteedd:: 33/16/9988 SamplesAnalyzed: 3/11-3/12/98 ACanlailbyrsaitsiboyn:Curve Rasge: ~~ E1SMp-S4p(3-)pbpb LooQfiCLaloiDb'ra:tionCurve: 0I.9m98 Call PHASEIV: Analysisofhistoric samples, 1970-805 Samp ppY+B4 s |! PrG7o-sSsEe1n3|] 13%3 20`.G0R1.A0H00809.0031] ] Esetr 30 30.0W1i.l0004000] apsosl] %2 30D-5e2m.0m510400011 . %RSD | Average (ppb) Std. Dev. | cress | [castbeation Croc cacy 110005%% EB | 5D 105% ] Branched Isomer Abundance Percentages ] `BrAanbcuhenddIasnomcoeerfs|||BraAnbchuenddIamnceoefs ampere| Avene 4. Dev) [@5eampiapeo)d_] | [isimvisduas Sampled in 1998 3%% % % 3MCotageGrovePlant Workers a " PoionldievdidSuearlsa SamSapmlpleeds in 1970's 31%% 5"% | 1-LOD/LOQ =LimitofDetectioonfQ/uaLntiitmatiiotn | stminamenao sos 004373 PHASEIV ] [= Poerv Ff j ar -- Se sT t Pafk Ooro = Prt2.0 Jn ar Bos ] E EIfemnae), =mnooouuom ow ows frIomommr Bm==eowBow ows | fn =2 I J 2=% = 1 =FEormati=n. = J 2= x H) wun , pIS jmrayn k m O E=E R ==z ow owes Nyen @mrTEmrn | fF jjoomnrsek E =B= E EE= wow | ffrrey FmA = ] ffIrood, reenns] == FBzA : E= wow | Rea LBiE: wows LJJIoaEeeEneE. A= 2m 8iou . : fH EJr] EEE3 wou Jo et [A [F Joamane B 2= E 2on wow: In 2B oa ou | 3SJ Lf1 aoaeMmeoa HutMk B3E==2 oE88 fJommsre as FEaa E wow an mr fI SNPooemewe1v31e99ops p"b "-2=--oo8wss ou. 8Poulet | Imm mu [ Te m-- ---n . . 004374 J ET / t i | 1 i| | | | | i || | |i KrisHaEnnsveinr-oSnrm.AenntaallytCLihacebmaoirsltatory- Fluoris FlBuuiolrdiinnegA2na3lEyt0i9calChemistry Team 612-778-6018 if emis Team Quantitative analysisofPPFhOVSaainnsidsVteIor:ic human sera samples and in human sera samples from rural Northern China a Summary: sMampales2rTi0w,nc1go9l9ashs8es.tvsFioaoflrssba(omMpt-lehecsstt),s,t,whseearfmierpsslutpewpvilotilehud1mt2eosvtaahmreipEeladevfsiirrnoopmnlm0aes.tn2itc0avl1iLa.la0sbm(bZLy-oJrfe)hfunMmadantndhseeeslre(ac3.oMnAMdvewodilituchamle) on. db`ceiotdmwnbeoietnnce0odn.at4nad-i1en.ext0nrmoaLuc.gtehodfvaeosalocuhnmeessfaaommrppellxeetw.raaTsctheixeotenrx;atncrtatechdteusssweiecnargseaeasnn,atilowynoz-pesadaiqrmuipanlngetriset(aaetg.iegvnetZl.ySAfioGxraspenrdafZiuZmoAriopSno)ctwlthaeenreeZe-sct A`snaunalaflloyynttaetaiend(edPnstRtiOafSnicd)abatryidomhnawigtahes-rvpierarelis.fsiuSerademblpiylqceuoismdwcpearrheisqruoanonotfmitmaaotlietvceulolyeagvrailrounaatHePpdLamhCgaarsiyenstss-epaaetlcsiptoeerncoticmaeelttlryyorpfr(teoHhpePaseLrxCeptd-r,EarfcSitaMveSedy:). aponianltyzeedexverty c$rsuarvmaep(lec+s2,t=we0re.e9w9di9t)h.iEna6c%hosfaemxppleectexetdrvaacltwueass.analyzedtwice.Calibrationchecks, c`haraccteori`sTutnhiseciipdnfregintmHiiaPtryLyroCifo-nmE,PSFaMOenSSdiMfndSisv.eecvIhenarraaallctrceaarsniedssot,iatclshyeesacenolaneldctyaetrdeyseaoxmhnpislb.ietseidnatchhiarpachtaesreis(tMi-csreetteoantliyo)nwtaimse, a PFOScon`cTehnetsreartaisoanmopfle3s.i4nptphbe;tMhesesta,mcpollleescrtaendfgreodmindoPnFoOrSsiconntcheenUtrSatiinotn hfre1o9m60151,.h81a0da59n.4a,verage CFhoirneaiganns`daTwhmeeprl9esedasemtpelremsiinnetdhtoZhasevte (aimnculcudhilnogw3ecromcopnocseinttreastaimopnolefsP)wFeOrSetchoallneacntyehdfurmoamndosneoarsdiantarsuertal . ```aqnumaalnteyiztetad(tp1ihorpneop.vbiWd)oi.utsThlhyte.hSreeicmxxaosicanpmtipinlogenso3dsfiaodmnnpeolhteucseomvnaitndaeisnnecPreaFdsOaPzSFaapOblSoecvcoeoltnlcheecentdteerdtaeitcnittoinhosenl1el9si8mi0ttsho(afWntihliplspa0bn0,a0lt2yh)te,itcmahleisteoaf7e theoalyhumansamplesdeterminedtocontainlessthan 10ppbPROS. Experimentalsummary: S.amplepIrenpaapraHtcioonnt:rIoolnl-epdaeiarivnigreoxntmreanctt,ioann ion-paiiagreagea,terabutylammoniumsulfate(TBA),is: atuaasarelgdeytttseoiPesxFttOrrSaaancstntfhderepraenedarl0fytlaenufoorn-ropmootolhacertmoara(tgnPracniOixaAc.tAs)one.liSvouenbnitscecqeotumhepynlotuatcnoedttasht,ee)lfiokdrermtiahetedi,coaanntdioofrnteihcceorneTsatgBieAtn-utat,ensdieiolnencmpteaitvieh,latynh,ael forMS analysis BHaPseLd:onCIthnahHreaPacLtfeCnr,iisaytniocaflrteihtqeeuanonttoiaolfnytttheiefmeoexrstftroharecsPtFasitOniSojneacrtye-dpahnadsepiansstehdtchorlouumgnhraeclahtriovemtatotoghrealpihquiiccdmoolubminl.e ehxaasmeplpaes,isinnagsttharnoduagrhdtsohleuctoiloun,mPn,FtOhSemanaayleyltuetseraetta1i0n.e5mdionrutaecsha.rRaectteenrtiisotnictaimmoeusbntoefttimwae.setFaoenrdanrd 004375 [ZY 1 i} HPPFOLSCssoylsutteimo.nandtheanalyte extractedfromsea nthisanalysiswerematchedtowithin 1%onthe | aEmSM,Sc:orADrueeatslepycotsniidosinonagfntPdotFmhOoeSnmsitataasnrsdioanfrgtdohsfeitnahdenicimaootnleiescctsuhularatfratchiteoapnnrti(zaCruyiSoOn)ch.arTahcitesriiosntiwcoafsPmoFnOiSliormedzsel=ec4t9iv9ely | tomaximizescositivity. Ascanof m/z=100to1210 (negativeonly)wasalsocollected. | `HcoPmLpoCuEEnSdS-i-MdeMSn/StMiMfSiS:ciaDtseivtoeenr.cytAsisioimnainlnEadSrmtMooSn,EitSaoMcrSi,hnegaxocfrecpahatcrtahtcaitteeoirntriisaidtdisssceslateenccitoaencdddd.aiArtfyotineaorlnsdsimeenlsieotnchoetfEcieSr-aoiMnnSyM',tSo - 1 scehcaorancdtaerryizeiosnticheriaognmfeuortthse(rdbauygshmtaesrhiionng),icthaapraarcttweirihshtiicgohf-tehneemroglyegcausl.eA,aSrearcerseualtteodaftnhdedeStmeacstheidn.g, For example,forPFOSanalysis, on 499isselecteda th characteristicprimaryion.Thisfon is sammaush(ecdiontrrotehesr iptoonsCnsFud3cSih0na,s)g,801a8m0aum(uco(rresCpondFingat.nodS2OS3)0,a0m9a9a5(mCyuF)e(ScOo,yrr)e.sEpoancdhinogfttohFesSeOs)ec,on1d3a0ry | fragmentsis detectedat the detector. || tQhuiaslsiatmyDecruoensatstrooonls,samumamptmlreaixrLsiympi:itkaetsiocnosu,lsdenrosbamepelveaslwuaetreednfootetxhtrsaacmtpeldeis.n.dEuaplcihcsaatem,pwlietehxt1reaxcctewpatisoann.alFyozred | ed ifnodutphliiscsaatem.plOeanreesbaamspleo,nMd0uOpl1i2c7a8tceaonnatlayisniesdoefncoaucghhomfattehretiawlofeoxtrtawcoteixotnrs.actMieotnsh,aTnhoelpbrleasneknstweedrreesults ono i be `spaomopleldeswearsapaurnchaaseedlvferyromyaS$zisgaemmaplwdeesr.eBperec-aeuxsterahcutmeda,nspsiekreadwwiitthhouatstPaFnOdSairsdcnoontcaevnatirlaatibolne,osfamPpFlOeSso,af d 1 rtpeho-eielxnittnreaexcattrearcdet.gTerdhecisuisropvrneeew-qeuaxasttarinaoacnltdeyedzrseiedvrbeaediffsoruronedamentrdhstaafovtoeedrttaohgbeeeosfavtemrhpyelsetiswm.oilbQarratcukoetthaiensgnacmuortpfvlteheisem(satta2rmipaxl=.eT0sth.i9is9sib9f)ai.soveednon | SpecificQCparametersar avaiableontheappendedresultstable. . | fLrionmitseorfa"Q;Tuthahenectmuirtearttehiaootndainalmyottiocpaltmiemtizheoddfboarsprbeeceinsuesloewd-tloeevveallu(<at5epp3ta)rqguearnatintgaetoiofn,PbFuOtSilnesvteelasdefxotrract-ed scaumrvpel,ewshiinletthheerraengieoasfa2i0s-t6ic0aplpdbi.ffGeirveeacaotfhoercsoanmcpelnetrsaetviaolnuscatheodsfernofmor5t-h1i0s0oppptbi,mitzhaetreion5onfotshteastitsatnicdaalrd -] deddesiffiefgennrdaeatnbecldeebu<enLttwiOelQeSanprspabm;epstltheiumssqatutiaendstvtiatclaoutneetdrafpirrnoesm1-e15o-pt5pspbthpebPl.FOiTSht.eIoqffqfuuuarntthiaetraatinodncofteocsrntiaohuintsbtdmrepecrateoachmioestdicso.tnaitySiiasntmoipclan elsly | 3 quantitationoftheIow-lvel2-stsamples (sample lessthan 3ppti required, a speciallow-level ] pLeakisohafm"pDTeehtiereeclstiutimloittninogffdretoemctPiFonOSfoarstahliyssainsa.lyPtFiOcaSlmsteatnhdoadrdimbaatseeidaolnaanndaPlyFsOtSreexcotgraaicttieodnforfotmhseedrisatinct raiansatolomyeozrfe.tdTbhheybetrhaaepncecoxhnoedfdicptaeicaohnkptreoeatpkhoiertnleidtnehhaeerHepP,eLsaCh-kotwrfaaceieralorycbccauosrenslsaiintteearasetpsdeoocliuwtfniiotcnorotehfpero1lbdourwcaisnbtclahenrededataernndt,di1o.1nl0ipinpeeba.raTPnhFdoOtshSee dLsiiasmmtipitlnoecfspqteuhaaankttasih)taabtpaieod,nwaPenFrOdeSdtlheeavesbl)isdegovannloua<taLttreOeedQacdt.tlStehaesmutpnhliaeqnstuhteehPaFltiOmaS)ihtpaoevfaqekuPsaFnhtaOipStela,etwivoeenlrasenedvdeasbli)ugaeatvaeiiddeaedntlc<eeLsdsOttDhh.eanthe `Thesesamplesare estimated to containfrom 0-1ppbPFOS. 004376 osiorss 2 i | FIPnsLtrCumenttealmspecifics: HewatPCacokalnudSmviae:sK11e0B0yeLsaiodnCeCshromatogaph | S2ux1mp00amrm csieze Fowrus: 300ymin | SoSSlovvleveneatntGA:r:atin:2M.e0thmaMnoAlmmoniumAcetate 1 4Ho5l%d5200%84 2.o9.c530.m5i0nmsi.ns. . FRoledn3.44555480oir13.55mi0.i 1 IInnjjeeccttiioonnvsoslaummpel:e1:02) | MEileccrtormoaspsMrsaPylmaatafsorssm2pI.eT4cSAstorPfotmMwyeaatrseersSpec"oChi,ck" | SCooneve alages: 2y 0,c6o0ve Soustemperature.+ 115C. | ATnoanlsy:ze36r5v,a4c1u3u,m4p9r9e,s4su2r7es: 0.006043 Bar, 0.000079mB Electrode! coseflow | | . . . B | i | i osonse 004377 3 } Fluorine Analytical -- | Chemistry Team 778-6018 | `Samples Received: 306198 `SSaammpplleess EAxntarlayczteedd:: J/11- 3/31029/09988 Analysis by: CaolfiCbraaltiibornatCiuornvCeaRravneg:e: LLooDQ*z:: BSMS (9) 1ppb- 49ppb. 039% S1pppbb PHASEV and VI: Analysis offoreign (rural Northern Chins) and historic sera samples (1960) |sper [GnomeJ sp|e "> r| p ppb+B40 | ZzZaAl|| L<OoDp 1 zzaa o<0qQ zas| za ma 8| oopD p Mm0o0o1r20s3]| MMooozn21o| Moo1208| 2 2448.44 4+--104 2112844--0021 59.4 +-0.1 | 2za7| <LoD ZZAZAu9maandn]10] <o0pQ ] QC Results [QC Checic | | Catbration Ce heck QC1) aRecovery 104% : ); J To| AT Branched Isomer Abundance Percentages |= -- Brasched lsomers| Branched Iiomers| : GiiDe) [@5ampitapeood] [StandardMaterial 2% | [[rInediCvoidtuaaglseSGarmopvleePd aint1r998ks a34s% | [iPnodoiievdidSuearlsaSamSapmlpleeds in 1970's 331%% | 21--LL0OQD==LiLmiimittooffQDueatnetcittiaotnion OR S198 : 004378 PHASE Vand VI { | Emme SOMME 1 | | yo tee 3603 | 008879 | [ioe , | /eighting: Null,Axistrans: None 1 ] I | ; | | - a| . Lo J | a/ | |i || !! 4 004380 4 !| KrisHaEnnsvenir-Sorn.mAennatlayltCihecmaislt tory- Fluoris ical i i FBluuiolrdiinnegAn2a-l3yEt-i0c9alCheaistry Team 127786018 Quantitative analysis oPfPFhOSVaainnsdhisVetIor:ic human sera samples and in human sera samples from rural Northern China ] SsuammpmlaersiyTn:wgloassestvsioaflssa(mMp-lsees),,twheerfeirssutpwpiltihed1t2osathmeplEeasviirnopnlmaestnitcavliLalasb(bZy-5Jte)fafMnadntdhesle(c3oMnMdewdiitchalS) on J Mbiaetdrwnceohetnc2o00n,.t41a9i-19n8.e.0nFmooLur.gbohfovtaohcslhtsse,afmsoparelmxpetlwreaavscotelixuotnmr;aeicvntaterhdiueessdiefcnrgaoamsnsi0to2nw-o1pa1mi.rp0imlnglreoa( fgheautmZ.aZSnAisSxrsasn.adZAmZvAiopSln)utwlmheeerZee-ssct | . sAcnuoalmlbfyiotnneiedd(sePnntFdiOfeSixc)tabrtaiycothneiwdgaha-sspvareenrsiesfsuiareedmpbllyieqc.uoiTmdhpcaerhiesxrtornoaocftmmwoaelretecauonlaalgryizoredaHqPapLtmCkiatsryasestc-pacetlci<yotnrlfoomieremptecerorytff(otHrohPreoLoeCcxe-tirEapaSncMstaEeSdy). i panoailnytteeanxdsttancrduarrvade(mcat2ert=ia0le.,9S9d9a)m.plEeascwhesraempqlueanetxittraaticvtewlay sevaanlaulaytzeeddatgwaiicaes.tCaaslpiebcriaaltliyopnrecphaercekds,, five: | a`nalcyzeod`eTvnhueesriiydfnegntsFiiaItmPyproLlCfe-smP,EwFSOeeMSrSieMdnwSis.etIvheinrnal6ll%rcaonafdsoesmxltpyheecsateneldaevlcaytletudeescsax.mipblietseid atchhairsapchtaesries(tiMc-rseetteontliyo)nwianse,8. characteristicprimaryion,andfivecharacteisicsecondary ons. | Fctrcsamples | PFOScon"cTehaestreartaiosnaompfl3es.i4nptbh;etMhessa,mpclolelserctaendgferdoimndoPnFaOrSscionntcheentUrSatiinonthero1m960131,.h8.a0d5i9n4a.verago ! Foreign"sTahmepl9seasmples ath Chinaandweredetermined 0Z-haevte nmcluucdhinlgow3ecrcoomncpelntiratsiaomnpolefsF)FweOrSetchoallneacntyehdfurmoamdnosneoardsaitrsuertal i analyzedpreviously.Six samplesdidnotcota : | mquaentitta(th1ioponp.bdW)i.tThhtehreeemxaciepntiinogno3sfaomnpelehsuemvaindesaecrPaesFdaPOmSRapOblSoeccvooelntlcheecentdteerdtaietcnittoihnoesn ile1ns9si8t0th8(aofnWti$hpilspa0bn0,a0tl2hy)et,fitchiaeltsooafrs theonlyumansampisdeterminedtoconta lessthan 10 gbFFOS. | Experimental summary: k 1 1S5a2mdpl1epeIxtnrraacptetHhpceoanantraIoorllnyl-peaadirerotaimvnitgirheoxenotmmraeantnctrt,ii:oxan.nAfnoino-npiacrcionmgproeaugnendt,,LtiektertabeutyClaatimomnoicnrieaugmenstu,lfsaetleeci(vTeBlAy),,is tfaanorarglMeyttSseaPniaFtlOryaSsnaissfn.edrrpeedtroflauooa-rpooloacotr(gaPanOnAoAas)eo.lSvuenbtse(qtuheynta1cttah)ef,odrimaetdi,oannofretchoenTstBiAt-uatnedioinnmpeart,hatnhcet BaHsPeLdCo:nCIhtanhrHeacaPtfLefCrini,isattyiaocafrletitheqcnutaointoaonlfyttthefoierxttfmrhoaercsPtetFiatOsiinSojneacrtye-dpahnasdepianstsheedcthorlouumgnhrealcahtriovmeattootghrealpihqiucicdomloubminl.e `Pehxaasmepplaes,isinngasttharnoduagrhdtsholeuctoiloun,mnP,FtOhSemanaaylyetleutie raetta1i0.n5emdfionre8sc.haraRctereisttictamieocusnbnteotwfteteineia.stFoaonrdnard osnmss 004381 ! V/ HPPFOLSCssoylsutteimo.nandtheanaly extractedfromserainthi analysiswerematchedtowithin1%onthe ESAS:ADneatleycstiisonoafnPdFmOonSisttoarnidnagrodsfitnhdeimcoalteecsuthlaatrtihoenprimaryioncharacteristicof PFOSism/z =499 taomym,axciormriezsepsoennsdiitnigvittoyt.heAmascsasnoofftmhe/za=ni1o0n0itcosu1r2f1a0c(tnaengta(tCiuveFoSnOlyy))w.aTshailssooconlwlaecstmeodn.itoredselectively 1 HPLCEESS--MMSSMMSS:Diesvteerctyisoinmailnadr m1o0nEitSoMrSi,negxocfecphtatrhacatteirtiasdtidcssaencaodnddiartyoinoanldsimensionof ertaintyto - | `chcaormapcotuenrdiziedsentthiefiiocnatfiuotn.hAersbiynsEmSasMhSi,ngaicthapaarrtwaitchthiiegohnr-ieisnseserlgteycitgeacds..AfAtserasreesullt oef thctehstemEaisSh-oiMnnSgM,,S 1 secondaFryoironiecxfarmpalgem,enfotrsF(dFaOuSghantaelryisoinss,)i,ocnha4ra9c9tiesrissetlieccotfedthasetmhoelcehcaurlaec,tearriesctriecaptreidmaanrdydieotne.cTthedi.sfon is s`maamsu(hceodriretsopootndhienrgitonosCsFu5cSh0a5s)8,01a80mam(cuo(rCr;eFsp.oSn0d5i)n,gatnodS20350)a,m99ua(mCFue(cSoOr5r)e.sEpoancdhinogftthFesSeOs)ec,on1d30ary ] fragmentsisdetectedatthe detector. | em tess metwr cris ptswit pen Ko QualityDcuonsttroolssaummpmlearliymi:tations, serasampleswerenotextractedinduplicate,with 1 exception. For ifnordtuhpilsiscaatmep.lOenareesbaamspeldeo,nMOduOp1l2ic7a8tceaonnatlayisniesdoefncoaucghhomfattehreitawlofeoxrttrwacoteixotnrsa.ctMieotnsh.aTnheepbrleasnekastweedrreesults | s`aamnapllyezewdpaesrainodailcyazleldytevoereyas$usraecmopmlpelse.teBiescolaautsieohnuomfatnhessearmapwliet.hoAumt PFiOSidsn(o2-t4apvplabi)laeqbulael,vistaymecpolnetlsroolf 1 r`peo-oelxetdascetreadp.uTrhcihsasperedf-erxotmrSaicigdmsaewreriseupnrdee-resxttroaocdtetdo,bsepviekreydsiwmiitlhaarsttoatnhdeasradcmopnlceenmtartartiixo.nTohfiPsRfOiSve,sid - | t`hpoeilienxetararcetgerdecsusriovneewqauastainoanldyezreidvbeedfforroematnhdeaafvteerrtahgeesoafmtphleetsw.obQracuketainngcoutrfvtehises(at+m2pal=0e.sti99sib9a)s.oedonn y SpecificQCparametersareavailableontheappendedresults abl. - ] fri Limit ofQuantitation `Thecurrentanalyticalmethodhasbeenused to evaluate a large ragofPFOSlovesextract-ed | fsarmopmlseerssi;ntthheemreatnhgoedoif 2n0o-t6o0pptpibm.izGeidvfeonrptrheecicosnecleoawt-rlaetvieolns(c<hoSpspebn)foqutahnitsiotpattiiomni,zbauttiionnsotfeatdhfeostrandard ] dciufrfveer,enwcheibleetthweereenissampstlaetissqtuiacanltdiitfafteerdenrcoefmo1r-s5apmpplbe.sTheveaqluatuedfaromn3d-ot1e0s0nipoptbtb,etchaeormeetisstniaotsistotaitciansltlyical ddeesfiegnndaatbeldeu<nLtOilQSaprpebe;stthiumsattheidsvtoaclounetraeipnr1e-s5aptsptbheFlOimSi.toIfffquutanhteirtaaticon foctauhnidsrmpreetachioscdi.onySianmples quantitationofthelow-levelZ-stsamples (sampleslesthan $ppb)is required,aspeciallow-level .LiofmDetiecttion | apneaalkyszhead`pbTeyhreetlshiuemlcitoitnondgifftdireotonemsctrPieFopOnofSrotarentdahlbiyesrsaies,n.aslhPyotFiwOcnaSlesmateratbnhadsoaedrliidsnmebaartesesreoidlaoultnaiaonnndaolPfyFs1OtbrSreeacxnotcgrnhaiectdtiaeodnnfodrf1otlmhiensdeeiarsratPinFcOt S riastomoeorf,tThehberaapnecxhoefdcpaecahkpteoathkeilnitnheaerHpPeLaCk-irsafcaierolcyccounrssiastteanstpdecoiwfinct,ortehpreoldouwcsibtlaenrdeatreda,ti1o.n0ptipmbe.aTnhdosthee dsiasmtipnlcetsptehaaktash)aaped,PwFeOreSdleeveslisegvanla<uaLtiOeedQad.tleSsamtphlaensthtehlaitma)ihtoafvqeuPanFtOiStalteivoenlasnevdabl)ueavtieddaencleedtsthheanthe l`iTmhietseosfaqmuapnlteistaatrieoensatnimdattheadttbo)cdoontnaoitnrreocmtt0-h1epupnbiPquFeOPSF.OSpeakshape,weredesignated <LOD, 05/07/98 004382 , | IHnPsLtrCumetnetaml specifics: `HewlettPackardSeries 1100LiquidChromatograph ] `Column:Ke2Byexts1ast0iol0Cnmiemg. FSloovwernattAe:: 3$200u0mnppMLairAmtmiicnml.oesniizuem Acca | SoSlovveennttGBr:adient:Meno! 1 45f109l0 %B.y125.e50minrs. nse. . [Hold at'$%Bf3o .5mir ns. 1|. InIjnecjtieoncv/otlsuaimmpoel:en1:0s2uL. | So Ser pte Electrospraymassspectrometer | `Micromas`sPlaMtfora2mI.4TASlaPtTwMyaarsesSpectrometer, "Chick" Conevolta--g20ve, -s60:v Sourcetemperature:+ 115C. Electrode! crossflow Analyzervacuumpressures:0.000043mBar, 0.000079mBar ] Tons: 369, 413, 499,427 . . . } TM- || | ose 004383 ! FluorineAnalytical jo Chemistry Team 778.6018 Samples Received: 36/98 Samples Extracted: 39198 SamplesAnalyzed: 3/11-3/12/98 | Analysis by: Calibration Curve Range: ofCalibration Curve: LoD': L0Qz: ESMS (9) 1ppb- 49ppb 0.998 1ppb Spb ! PHASE V and VI: Analysisof foreign (rural Northern China) and historic sera samples (1960) |co|nn| 1 zn] oo p ZA (ppob+DB40) | za 200 24z3m)| <aLo0qQ | ' ZZA6 and 3 <LOD sToOOoToS]] MMoMooOot1za22o01s2|]} e24v42i+1l-0s% 312513482+444--/-000112 | 2zZAvmzaztao7|| <<LLo0DQ ZZA11 and 12] <LOD QCResults ] QC Check: Catbaton Check (C1) | CalibrationCheck (QC2) % Recovery 100% 106% ; _ ; "x | | BranchedIsomerAbundance Percentages : | Adameof| 5Avuaduncoef Eampieror Branched Isomers|BranchedInomers 1 erage) Gabe) JGaremptispoeod--] | [StandardMaterial [3inMdiviCduoatlsageGSraovmePplialnn1te9W9od8rkers % a34% | PInodoilveidduSaelrsaSampSlampeleds in 1970's 3%% 21--LLOOQD==LLiimmiittooffQuDaentteicttaitoinon : 3M Environmental srs Lab 004384ppv umavi | EEmmEmae ETOMAAME ] Co | abe 3643. | 004385 | ---------- TT Esrnian, iGim i g--J-- ationca oefcint =0.999912, 42. 0.999823 CT Fic gi: cite,Welghting Null, Ais rant:None 2.0985. i i ! , || /Joe 1 | | |: |i / | | ! i% 5 To fae . TTR | | | | | 004386 | 1i Kr3iMsHaEnnsvenir-Sorn.mAzeanlyttacall Laboratory: Chemist Fluorine Analytical Chemistry Team BFuliuolrdiinnegA2n-a3lEy.t0ic9al Chemistry Team 612.778.5018 i' : US Geographical PhaseVII: Distribution Stud-y PFOS levels in human sera | SummarFiyf:ty-fivesamplesof oldhumanserainplasticcentrifugetubesweresuppliedtothe ] `EcwaoevlrlioerrcoetnemmdoefvaretoamdlsfiLrtoaemsbacbrayocJuhensdfatMmhapenlUdenaeinltde(3edMSxttMraeatcdetie,cdcawolin)ttoahinanMneadsireocnvh-ep1raa9ilraimnngLdreMoaafgrescnertha;.n2e0Oa,rn1le9y9m8a.lLTaohflethsiasqmapumolpefolsset,esrswa:ere 1 (eBxFtOraSc)tbedyihnidugphlpirceastseu.rTehleiqYuiad cexthrarctsowmeraatnaolygzerdquaanptihtmaatysivs-eslp<yefcoltrpeoemrecfturtyorr(oEoocPtLsaCnp-esEruSlMafSoy)na.teAnalyte iadenendtxisftiatcnadctariurordnmvawaeat(esc+rvi2era=tilf0.i.eeS9da9b6md)yp.cloeAmspmwaaertriresiqoxusnanpotifikmteoacltaeilcvieubllryaatreiivoanolncuhaHtePecdLka,Cgaraneiatnlesyntztaeisodpinencttiiahmleelmyafiptrdhedeplaeerxoetdfr,atfchiteveaedn-aapnloayilnstyitse | sequence,waswithin1%ofexpectedvalues. | bFrpancthheeTdscahhmeapslieenrsiarssaoanmmgeperlwdeaissinnP4tF1hO+e/Sg-ceo4on%gcreanItpnrhaistctiaoinnsdtfairrbdousmtoil9ouptnpsibotnusdo1yfh5aP9dFpaOpnSb,.atvTheehrbearpgaeenPrcoFhcOeaSdtcachobanicuennnatcdrcaoatouinnfotctnshofeefo2r8 217+/3.1%ofthetotalconcentration. Adetailed summaryoftheresults iatached. : | Te tyo EOS resin cst plein is ae(en c`haraccteorisuntsicipfnrgiHmiaPrLryCi-onmE,SaMenSdMdfSi.veIchnaaralcltcearsisetsictsheeacnoanldyatreyexahnisb.ited acharacteristicretention time,a Enma : | SamplepIrenpaarpaHtcioonnt:rIoolnl-epdaeiarviingroexntmreaactt,iaonioo-pairingreagent,tetrabutylammoniumsulfte(TBA),is | e (`PuOseAdA1)0e,xatrreascetltehcetaivneallyyttaerfgreotmetdbhyetmhatrciaxt.iAonniicorneiacgecaot.mpSouubnsdesq,uleinktePoFtOhSeafonrdmpaetrifolnuoofrtoohcetTanBaAs-taenion a eget ermineTA `mefortHFhLC-aESMnSanoalylsis. | BHaPsLeCd:onCIthnahrHeaaPcftLfeiCrn,iistatynoiacflriteqhtueeaonnttaoiolfnyttthieemeeoxsrtftroharcePsitFsaOtiSinojneacrtyed-aphnaspeianstshedectohlruoumghraeclhatriovmeatttoghrealpihqiucicdmoolbuimlne.- | PpChFaaOusSeppslaoelsusntiinsogntsahanrnotudghehedtashnelaclaoyitleunme,nxPt,rFtaOchtSeeamdnfaarylodymtsetiesrari1ent1ta0hini5emsdiafnotarleyassci.swheR arermaatcci htiaeemdnotsrouwbiniete tstohwftieinei1mne%c.on anFontrhie d ' HPLCsystem. `EamSyA,Sco:rArDneeastlpeyocnstdiioisnnaogftnoPdFtmOhoSenmstaiastnsdooafrrtdoifhstinehnagedniimcooanltieecctsuhularatfrtachotenapnrti(mCaerFyirfSoOn)ch.arTahctiesriiosntiwcoafsmFonOiStiorsemd/szele=c4t9iv9ely tomaximizesensitivity. Ascanofm/z=100 t 1210(negativeonly)wasalsocollected. | 004387 05798 1 | HPLCES-MS/MS: Detection andmonitoringofcharacteristicsecondary fons i `compounEdSi-dMenStiMfSicaitsivoen.ryAssiiminlEaSrtMoSE,SaMcSh,eaxcrepatcthtaiteointraiiddsseslatenciatdecddi.tAifotnearl deimlenesiocnthotefcEieSrot-aMinnStMy,Sto chsaercaocntdearriyizoensitchfrfaognmfeunrtthse(rdbayusgmhtaesrhionnsg)i,tcahpaarrtawcittherhiigoshft-t<ihnceermgygoas.lAsareecrreecssutletudoaf nthldedsemetaescht,eidn.g, smashedFinotrocoxtahmepriieo,nsfsorucPhFOaSsa0naalymsuis,(cioornre4s9p9onidsienlgec0tSedOa),th99cahamruact(ecroirsrteiscppornidmianrgyitooFn.STOh)i,si1o3n0is afmur(caoirgdreteemcstepdetaootnCtnFhdeSdiOte5nt)egc,sto1r80amu (GSO0)d2,30 amu (CSFeSOy). Eachofthesesecondary `Quality cIonnmtorosltscuasmemsa,rthye:serasampleswereextractedinduplicate;forthosesamplesthatwere,standard | deIviaavatiiloanblvea,ltuheesmaarteriinxclsupidkeedirnectohveertiaesblaereofirnecsluuldte.Mianttrhiexrsespuilktesswtaebrlee. pArempariedfdor-s(e4vl9epreaplbsv)amqupealleislt,y oo. sceoznatwriotlshaomuptlPeFwOaSsiasnnaloyzaevda;ilraebceo,vsearmyfploerstohfempaotlreidxsspeirakpeuwracshdaesteedrmfrionmeSdtigorbmea w1e0r1%p.reB-eecxtaruascteodh,uman ] sbpeivkeerdywsiimtihlaasrttaontdhaersdacomnpcleenmtraattriioxn.oTfhiPsFOfiSv,eapnoidartee-xetxrtaracctteedd.cuTrhviewsapsranaely-zeedbxesfeortreiras nuanddaecfrtsetrttoahodetdo | Osfatmhpeltesw.oQburaanctkiettaitnigoncourfvtehse s(a2mpl=0es.i9s9b)a.sSepdeocniftihceQlCinpeaarrarmeegtreesrssiaorneeaqvuaatiiloanbedoernitvheedfaprpoemntdheedsrveesrualgtes: table. | LiofQmuantiitattion "Thecurrentanalyticalmethodbasbee use toevalua alargerangeofPFOSlevelsextracted f`sraommpsleersai;ntthheemreatnhgoedoifs2n0o-t6o0pptpibm.izGeidvfeonrtphreecciosnecleoawt-rlaetvieolns(c<hopspenbf)oquratnhtiistoapttiiomn,izbauttioinnstofeatdhfesotrandard ] cduirfvfeer,ewnhciebleettwheereen issaamsptlateissqtuicaanltdiitfafteeredfnrceofmor1s-5apmppble.sTehvaeluquaatnetiftratoimon 5d-o1c0s0npoptbb,tehceormeeisstnatoissttiactailsltyical dedesfiegansaitbeldcu<ntLiOl Q5appebe,stthiumsattheidsvtaolcuoenrtepares1e.n5ptsptbePFmOiSt,oIffqfuuarntthietraatiocn focthauinsdmrpertehcaoisdic.onSyainmples | qQuantuitaationnsoyifstthteemlwoaiwllltebveeirleZq-ousierneds.amples (sampleslessthan ppb)isrequired, speciallow-level ] pLeiamiktshoaf"pDTeehtreeecsltuiilmotinitnogffdertoemctFioOnSfaontaliyssainsa.lPytRiOcaSlmsettanhdoadrdimabtaesreidaolnaanndaPlyFsOtSreecxotgraaictitoendofrfotmhseedriastinct ] ainsaolmyezre.dTbhyetahpeecoxnodfitciaocnhspreeapokritnetdhheerHeI,PsLChHorwanceeaorcbcausreslainte resspoelciuftiico,nroepfro1 dburcainblceherdetaenndti1olnitniemaeraPnFdOtShe `astaimopolefstthheabtraa)nbcahdedPpFeOaSkleovtehleselvianleuarapteedaakisefsisrtlhyacnotnhseisltiemnittdofoqwuaanttoitthatlioonwanstdanbd)aervdi,d1e.n0cpepdbt,hTehose. | dliismtiitnocftpqueaantkisthaatpieo,n waenrdetdhaetsb)idgonnoa<ttLeOleeQdc.ttShaemupnlieqsutehPaFO2)ShpaevaekPsFhOapSel,ewveerlesedveasliugantaetdeadtl<eLsOstDh.anthe , Thesesamplesareestimated 0contaia from0-1ppbPFOS. i HIPnsLtrCusmyesntteamlspecifics: Hewlen CPaoclkuamrdnSKeeryiessto1n1e0B0eLtiaqsuliCdiCshromatograph 2x100 mm $ jum panicle size: 004388 oss " || FSolvoewnetAr: 32000MMmAimam.onium Accte + SoSlovveenmtGBr:adient:Methanol H4old5at5%108%iB0nfo9r.9530.m5i00nmsi. s. RHeotludrnatto434%584fBor133.18m0inmsi.ns. . Injectionvolume: 101. Injec/tsaimpolen:1s `EMilcroemacsstPlmartafsoosrssmpepc1AtrrPoImaMetayetr Spectrometer,"Chick" | C`onMevaolst2.aL4gSyeo6fs0tn:vwaxre MSoodues:ep egutviesc. ] ATEnolauelscy:rzo4ed9er9v:,ac4rc2ou7susmporewssures: 0.000043mBar,0.000079mBar | . . oss 004389 3 Emm Fluoriine Analyti&cal Chemistry Team SR [ om BE I ets mi-- 3a8 PEASEVIReus:USgeographical diiron dy - PFOSlevel CPAaleCi:naCavcaansge Lon 100% bums era E Naa oxo nosw | oFNwo En boam oomm Bomo oonu 1 GSSaomoe mMuosrop EI | Teboomes 3A2 m00a1 | Won 8 01 r muE 42 ] btheoee sB BB n1s | oa=Wso on ow m ra mn0s YwYoooorw u w3u a8es | eso 3s 83 QCResults ] fE eos ee] oo] [IE aanRxom Teas W2Naowio a318 NNmoonon 8da wNNeemu m n 9 5 o20 WwWoien m ua o2a0 Wwee 3noeor mmBosonG8nooosz i= prm :x 5is wDTmemx oaiuars WAseo3 or | rCirooleSGreomtPt198at, 1998 =Toon F~=H =53 | [Ff oiieesSe ei,a5re s 350 prnppriy fi~t ~ pF-oo -ii x 3 Sop oe ho po 5 31+L01D LLik0nk f0enii"nin ss _-- Traeats mFineabyudam Tob Er . - .: . 04390 PHASE VI EL ee BESTCOPY AVAILABLE. ~~ | EEE EEL pLi feos 004391 !: E--E---- pEm--pm--E BESTCOPYAVAILABLE = | | || E EEEE | |i ! ~ 1 | . | . |[Fee N| | } . Ii - Po / | / lCl er . . | 00439. | | 3M EnvironmentalLaboratory- Fluorine Analytical Chemistry Team Paeisn s33805 KrisHansen -Sr.AnalytCihecmaislt FluorineAnalyticalChemistry Team `Qua`nhtuitmaatnivseeraanaslaymsipsleosfPfFroOmSSiwnehdiesntoric i | SNuaamnmdaerdm yC:oogeeess.rsampltoestrTospaiecoovnfiolpwpiees(iseppplosd5.oi1hSoE5dv5ei.n1o10mten1nSaaEl1lL1e-abSE,by2J)Tle 7 eiet. Tacs we syedately tps volumevariedfrom0.5to 1.0mLofhuman sera.Anexactvolume,usuallytheentiresamplewas lfosate(PFO)by highpressureiid chromatograpmahsys-pe<ctlromeetcryt(HrFoLCs-EpSMaSy). | "Analyt identification wasverifiedbycomparison ofmolecularionEPLCretentiontimeoftheextracted | eesti pointextractedcurve(2 =0.992). `Eachsampleextractwasanalyzedtwice.Calibrationchecks, | Descripti`oSenroafssaammpplleessSgEr-o1uptsh:roSuEg-h1StEo-S1E0-w1e0raencodlSlecEte-dtf1roo2m10Swedish donorsmore recently than `samplesSE-11throughSE-20.`TheaveragePFOSconcentration inthefirst groups wasdetermined to be 1.3ppb;therewerethreesamplesbelowthelimitofdetection. "Thesecondsetofsampleshadanaverage ,| SEE xapmeprliemperinepteraaotmoasnm:maoryv:paoafrinieosn.eoes A xoewecotovcnapc ep,lF.OScPi oHOn%csstaco: nad wi. cpcb.mT; 1iw bpcosi seto ] TctfhorMaSnanaoislis | Ina pH controlledenvironment,aniou-pairing reagent,tetrabutylammoniumsulfate(TBA),is: used toextract theanalyte fromthematrix. Anioniccompounds,like PFOSand perfluorooctancate (POAA),areselectivelytargetedbythecationicreagent. SubsequenttotheformationoftheTBA-anion pair,the analyteistransferredto anon-polarorganicsolvent (ethyl acetate),dried, and reconstituted in | PLC: raceres entonteestfo PtFyOScid paseo chromsiograpiccls. `Basedontheaffinityoftheanalyteforthestationary-phaseinthecolumnrelativetotheliquidmobile- `phasepassingthrough thecolumn,theanalyteisretainedfor acharactaemorunitsofttiimce. For | example,in a standardsolution,PFOSmayeluteat 10.5minutes.Retentiontimesbetween astandard. a rtEo srsayswerahd wii 190 HPLCsystem. ,; . - 'ES/MS:D"eAtneacltyisiosnoafnPdFmoOnSisttoarnidnagrodfsitnhdeimcaotleescutlhaartihoenprimaryfoncharacteristic of PFOSism/z = 499 | ano 13 51 peo wssl coll. I hosesie ! `am,correspondingtothemassoftheanionic surfactant (C4Fy;SOy-). Thisionwasmonitoredselectively | awnoms 004394 1 thatweredetermined tcontainPFOSat concentrationsgreat than 10ppb, is possible toverify the | identityoftheanalyteusingHPLC-ESMSMSto monitorfour characteristicfragmentdaughterions. QualityDcounetrtoolssaummpmlaelriymi:tations, serasampleswerenotextractedinduplicate.Forthissame reason, ``mMaettrhiaxnsoplibkleascnoksuwledrbeoatnbaeleyvzaeldupaetreidofdoircatlhleystaomepnlseusr.eEcaocmhplseamtpliseoelxattiroanctowfathseaszaamlpylzee.diTnwduoplmiicda-tlee.vel s2er.a98wpitphboauntdP4F.O9S5 pipsdn)oqtauvaaliiltaybcloen,tsraomlpslaemspolfepsowoelreedasnearlaypzuerdcbheatswedeferneovmeSriygmsaawmeplreesp.reB-eecxtaruascetheduyman |i bspeivkeerdywsiimtihlaasrtatntdharsdacmonpcleeatmraattrioinx.TofhiPsFOfoSu,rapodotre-eexxttrraacctteeddc.uTrhviewspasranealy-zeedbxesfeortreiraunanddaecfrtseitrotoehdetdo | osfatmhpleetsw.oQubarnatcikteattiinogncourfvtehs s(a2mp=l0e.9i9s2b)a.sSepdecoinftihcQlCinpeaarrarmeegrteesrssioanr eaqvuaaltiaobnidoernivthedafprpomentdheedarevseurlatgse. able. 1 LiofTQmhuaenstieiltaottwiolnevels~ampleswereanalyzedagainst aspeciallyprepare lowlove curve.Asaresults, `econvfidaenlcaetlui0maaitnsdt1caelpcpudbl.ateFdoarrtohuinsdtanhaelycsuirsv,tehvealluiestinodfiqcuaatnteiataqtuiaonntiafinabtlheedilfifmeirtenofcedbeetetcwteieonnasraemples I Liofn"DTehtdeelcittmointofdetectionforthisanalyticalmethodisbasedonanalystrecogaitionofthe distinct | p`aenaaklsyzheadpbeyrtehseulctoinndgiftrioonmsPeFpOoSrtaendalhyesries,.shPaFwOnSesatrabnadsaerldimnaetreersoilaultainondPofFO1bSreaxntcrhacetdeadnfdro1mlsienreaarFFOS 1 raistoimoearf,tThehberaapnecxhoefdpceaachkpteotahkeilnitnheearHpPeLakCi-srfaaierloycccounrssiastentspdeociwfitcortephreoldouwcibsltearnedtaredn,t1io.n0tpipb.gTohdotshee i stahmepulneisqtuheaPtF)OShpaveeaPkFshOaSplee,vweelrscedvaelsuiagtnoadtaetdlbe.ds.tL.h(abnetlohwdleitmeicttoiofnqulainmtiitt.aTthioensaensdthaatmb)adproenelsottiermeaftlseedc.t 0coen ssthatn1pipb Pn FOS. ouzorss 004395 2 HIPnsLtCrusmyesntteamlspecifics. `HewlettCPaoclkuamrdSeKreiyessto1n1e0B0aLsiqiuliCdiCshromatograph 2S yxm1p0a0rtmicmlesize 1 FSloiovwenstteA:: 320.00miLMiAmmimn.oniumAcetate | SoSlovlveeanttGBr:adicat: Methanal | 4H5o%015Bi0nd9.5530mi00a nmsi. FRoetludrant4o54%5%8fBora31..55m0imnis.ns. 1 IInnjjeeccttiioonnsvolsuammep:le1;024 | MEiclroemacsstPlamrtafsaosrsmspeIpcTAtrrPoImaMetayesrs Spectrom"eCtheirck," ``MCaosnsevLoylntxag2e.s4:So-f20tvw,a6re0v . | MSoodurec:eetleemcpterroastpurraeyn:egativ1e15C. `Analyzervacuumpressure:0.000043mBas, 0.000079mBar | Tons: 369,413, 499,427 | MEileccrtormoaspsrsaQyuMatSrMo ISl APLMSMS,"MadeliTM C`onMevaolsta3sg.es1L:SoSyf0tnvwaxre : | MCooldlies:ionelgeacstreonseprrgayyn:e4g0ative Y4 Sourcetemperature: 115C. : | APnarlyizemrJvoaua:cr4u9uym9pressures: 0.000043mBar,0.000079 mBar : 1 El`ectDrodeai:oZnusa:og0u,5s9h,13i0,1e80, r230 | unos 004396 3 i Fluorine Analytical go. Chemistry Team 778.6018 SumplesRecelved: ass SSaummpplleessEAxutlryascetde:d: 44075440385588 prema CaPlflrCaatlilobnraCiuornvCeuRravseg:e: Lon': 10Q* `Study of PFOSlevelsinhistorical Swedish humansera samples Es) 01352010 100350 mb mb | Ee | me gr | awe SEET2 T12540011 S= ET T29144-0070 sSBE4s 10b4a-l01 | sEe1i4s 228944--0000 | ss8E76 218244.-0000 sEe1167 212744-0020 | SsEEerSsr. 26ba4av-l09 ssFeeiss 12bw4au-l00 QCResults bbrraattiioonn CChheecc24.99kk85p7,,3p0b i bbrraattiioonn CChheecc42..99kk58pp,,ppbb : bration Check, 2.98ppb | bration Check. 495 pp 10To0o%% 10906%% : ssoa ] `Branched IsomerAbundancePercentages : :2 : . [Sfnivaicualns daSrdMeatemriapl iln15e58 oo%e | f[nividunl rSeaGroc mipo itn1l9wm7o08keseed,d10n558 aasm | [[FPoocrteedigmnSeSirearSStamepleesa,firo1pm98149e:s97d.s159854 ao~am US Geographical StdySamples, 1598 an | 21-ILTooD=QLiLnaitkofoDteutnecittioinon ED wh E"A 33 PEAY 2n : " 5 | Pagel | 004397 . Sheet! . filo number 4009807 sam0p.l9e98dpepsbcirnijpt1ion pe4a6k5a4rea [ana0l8yte] 44009999880089 229988ppppbbiinnjj21 88204976 3312 44009099881101 44.9955 ppppbbiinn12jj 1182881 6645 4099812 9.8ppbinj1 us 102 144000099881138 9.8 ppb inj 2 Mech blank mee 99 4466 05 [PFOSJavg St.Dev. | 44009099881290 SSEE--1111HHSSMMOOTT88iinnjj12 99266559 1329 a 02 | 44009099862212 SSEE-1122HHSSOM4O0T7B8iinnjj12 77878964 2390 29 [3 4009823 SEI3HSMOBinj1 6507 20 ] 44009009882254 SSEE1I43HHSSMMOOBTi8nmjj21z 67763864 2218 20 01 ] 44009090882276 SSEE--1I4SHHSSOO0U7T88iinnjj12 77687048 .2289 28 00 4009628 SEISHSMOBinj2 7808 4099829 Mech blank 4183 2093 29 00 %Rec,Q:C | 44000099883301 2.4998ppppbbccaallcchheecckk 180177231 6802 108 100 44009099883323 SE-MeIcHhSMbOlaBniknj1 87874646 2090 | 4009834 4009835 SSEE-I167HHSSMMOOT88iinnjj21 76451319 2168 27 02 : ] 44009099883367 SSEE--1I8THHSMS4M0O788iimnjj12 55435800 1122 12 00 44009099883398 SSEE--I19BHHSSOM0OT"88iinnjj12 54547599 o1s2 12 00 - | 4400999934401 SEI9HSOA0TBin2 SE-20HSO0T8inj1 4335 4018 04 oz 05 or % 4099342 SE20HSOA0TBinjZ 8971 01 02 i 00 | 4049090884434 29M8eppobhcballacnhkeck 87596892 03.01 : 100 ] 4009845 4009846 A9ppbealcheck Mech blank 18014873 0483 % 4009847 SE1HSOA0TBimj1 630 11 44000099884489 SSEEI2HHSSMMU0TT88iimnjj12 55473768 1124 12 01 ] 4009850 4009851 SE-2HSO0T8inj2 SE-QHSOA0T8inj1 5G18987 1105 15 01 . | 44009990885532 SSEE--4QHHSSOMO4A007T8Biinnj12 G581347 1110 10 00 44009999885554 SSEE--S4HHSSOMAO0TT88iinnjj12 6127 4807 10 01 10 01 |! 44009990885576 ~~ SE-5HSOM078imj2 Mech blank 4987 a6 08 08 08 o1 : 44000099885580 429958ppppbbccaallcchheecckk 7479 9904 27 11 8 FE] 4099860 `Meoh blank 219 04 004398 Page 1 Sheet! 44000990886612 SSEE6GHHSSOM00TT88iinnjj12 4542810 1122 12 00 44009990886634 SSEE-7THHSSMOO0TT8iinnjj12 77617289 2288 28 00 4099865 SE-BHSM0T8inj1 4566 08 4090866 SE-SHSO078inj2 4682 Ig [3 01 i 44009999886678 SSEEOOHHSSMO00TT8Biinnjj12 68419873 3220 26 09 . t 44009909887609 SSEE--0IH0SHMS0OTM88iinnjj12 44371120 0041 0s oz . | 44009099887712 watMeercbhlabnlkainknjl 13685153 0000 44009999887734 wsaetrearbbllaannkkinijnj12 4881931 0014 1 44009099887756 s0e.r9a8b8lpapnbkiidnjj12 54419%8 0150 4009877 0.998 ppb 2 5054 09 i 4000878 2.98ppbinj 1 m2. 28 - `44009090888709 24.9968 ppppbbiinnjj 21 70129927 2479 | 44000999888821 94..8905 ppppbbinnjj 21 01680576 4994 4099883 9.80ppb nj 2 1638 94 i . 1 004399 |i Page2 1 Kr3iMs HEannsveinr-Sorn.AmnenatlayltCLihaecbmaoirsltatory: FluorineAnalytical Chemistry Team FBluuiolrdiinnegA2s3a5ly.i0c9alChemistry Team 612.778.6018 , | humanQsuearnatistaamtpilveesafnarloymsitshoefUBniFvOeSrsiitnyhoifstMoirincnesota .| | |... | | Sommar"yT:ensamplesweresupplied theEnvironmentalLabby JefMandel(3MMedica)onApi 14, l61eA9v9,e8ls.7.ATT,hh8ee,ts9eeAnt,easnnasdmapml1pe0lsAwe.sTewoeepr-aeorcntloeafsmsLaig.fripeoodrutspi"oolfnosswoa-flmephvleeelss"tamh(pa<l1,e0spfewbre)r;exttchrxayicrtaaicrotene,duwmesbrieenrpgreaed-1si,co2rneA-e,on3iesAdif,nog4r.,P5,EOS prreeasgseunrte. lTihqueiedxctrhactrswoermeaanatlyozegd qruanitiptamthaivsyelsys-fpoe<cptlerrocfmletotrryoro(coHtaPsnLeCps-uErfSoaMnaSyt)s.(APnaFiOySt)euisdienagifhiicgahtion `mwaatsrvieli.fieSdabmypcloemspwaerirseoqnuoafntmioslievceullyacrailonu-aHePdLaCgarietnesnttaiopnetciimaleolyfptrheepeaxrtrea,cftievdeapnoailnytteexatnradcstteadncduarrvde (20.987).Calibrationchecks,analyzedevery samples,werebetween80-108%ofexpectedvalues. SEDescIrniaolpflsttaemnipfloetsnh:esepre-screenedsamples,PFOSconcentrationwasdetermined obelessthanthe detection mitofthemethod, 1pps | uSsaemdptloepcxirracethcpeaonaatloryntpeaafierrotimngtiaheexotmratatncrtiix:o.nAoninoenircicnogmpaaugnednst,tikebPFlOSsandpierfloriooctanCcaOtAe , (pPaOiA,Ath)e,anaarleysteelecsttirvealnystfaerrgeede0db2yntohne-pcoaltaironiocrrgeaangiecsnotl.veSnutbs(eeqthuyelnatctao it)h,e fdorremdat,iaonnodfretchonTstBiAt-utaesdifonn metfhorMaSnanaolyslis. | HPLC:CIhanrHaPctLeCr,isatincarleitqeunottioonftm heextefroarctPsFiOnSjectedandpassedthrough chromatographiccolumn. Base ontheaffityoftheanalytefothestationary-phaseinth columnrelative to theliquidmobile- - | phasepassingthroughth columa,theanalyte ireainedforcharacteristicamountoftime. For `example,in a standardsolution,PFOSmayeluteat10.5minutes. Rettimeesbnetwteeniastoandnatd PHFPOLSCssoylsuttieomn.andth analyteextractedfromserainthisanalysisweremalchedtowithin 1%oathe .ESAS:ADnetaelcytsiionsoafndPRmOoSnisttoarnidnagrodsfitnhdeimcaatleesctuhlaatrtlhonprimaryioncharacteristicofFFOSism/z = 499 | `a0mmya,xciomrriezsepsoennsdiitnigvit1yt.hAe msacsanoofftmhz=a1ni0o0nitcosu1r2f1a0c(tnaengta(tiCveoonOly).vaTshailssioocnolwlaesctmeodn.itored selectively Q`ucoamlpilteytecSoiesnrotalrastoailmonpmolaefmstmwhaeerrsyea:mepxtlreaactnedditnwdoumpildi-claetvee.lM(e2t.h98apnpoblbalnadn4k.sw95epreta)naqulaylzietdycpeornitordoilcaslalmypltoeesnwseurree apnoaollyezdesdebaeptuwrechnaesveerfyro$smamSpilgemsa.wBeerceapursee-ehxuramcaends,ersapiwkietdhowiutth PaFsOtSanidnarod acovnacielnbtlrea,tsioanmoplfeFsFoOfS,and reextraciod.Thspre-esractedser sunderstood tbevery similar thesamplematrix.Thisive 0422/98 004400 1 i | | | |. | | | | | | ptohienlitneexatrrraecgtreedscsuirovneewqausaatinoanldyezreidvbeedfforrosmatnhdefaeverrtahgeeosafmptlhest.wobQracuketaingncoutrfvtehise(sat2mpal=e0st.i9s8ib7)a.soedonn SpecificQCparamtersaeavailableonth appendedresults table. Limitof`QThuesaelnowtleivLetilmsiaatmotpflDieestwoeecrntei!aonnalyzedagainst asociallypreparedlowlevelcurve. As aresults, ceonfvidaenlceatul0imaaitnstdca1elcpupdlba.teFd oarrotuhnisdatnahlycsuirs,vtehvealluiestinofdiqcuaatnetiataqtuiaonntifainabtlheedilfifmeirteoncfebdeettocwteieonnasraemples equivalent. HInPsLtrCusmyesntteamlspecifics: HewlettP`aCcoklaurmnd:SKeeryissto1n1e0B0eLtiaqsiuliCd Cyhromatograph p2mx10pa0rtmicmle size FSoobwvenattAs: 23.00mmMAimnmoniumAcetate SaSloviveeaattGBr:adient:Methanol : 45%1B10n 9.950mi0ns. H`Roeltdurantt9o04%54%BfBoira3.15500mmiinsn.s. `Toldat 45 % for3.5mins. IInjnecjtieocnvt/osliaumompenl:es:102.iL. 4 . . `EMilcroemacsstPlmaratfsoosrssmpeIpcAtrrPoImaMetaeysrsSpectrome"Ctheicrk," `MConaevsoltsa2g.Le4sS:yo-f2tn0wva,xr6e0v MSooudrec:eteelemcpterroastpurraey:negativ1e15C. y `TAosnasl:y3z6e9rv,a4c1u3,um4p9r9essares: 0.000043mBar, 0000079mBar i| Electrode:crossflow unas 004401 2 Fluorine Analytical Chemistry Team `Samples Received: SamplesExtracted: 44s 4nsm8 ! Samples Analyzed: ~~ : 4/164/21/98 Analysis by: CalibrationCurveRange: Loopf*C:alibration Curve: i AnalysisofPFOS in historic samples of human sera(ULoofQM*) aE Samp ppb ] 21 t<LooD A 4 <<1 Lop lols <10D e . T=" ] wA <oLooD osA |. <<L1op EN <10D Hansen 7786018 ESMS () 1ppb 1024.9ppb 0L.o98m7 ppb . | QCReus . o I 3 [Cm ea eg] |CaliCbhercka(QtC?2iato4.n95 ppb) 108% - | 1-LOD = LimitofDetection * i 2-LOQ = Limit ofQuantitation anus 004402 KrisHaEnnsveinr-Sorn.AmnenatlayltLCiahbecmoairslattory-FluorineAnalytical m FBuliolrdiinnegA2s3al5y0ic9alChemistry Team 61778.6018 PFOS levels in individual's humanPsheraXaIs,aPsmaprltee2s:, a blind repeatability experiment | SummarFyi:fteensamplesofhumanseain plasticcentrifugetubesweresupplied 0theEnvironmentalLab tbhyaJ,eaffMeraenxdteralct3iMonM,ewdeirceaplr)e-onscArpereinled1f5o,r 1P9R9O8.STlehveeflisf.tTeehnessaemfpliefs ewesraemppalretswoefraegcrloauspsioffiesdaamspl"emsid- ] le12v,ealnd1130.pSpabmaplnedo<f1s0e0rpapfbro;mtthheeysaerdeonrourmsbhearvedeb1Ae,e2nBa,na2l,y4zAep,reAv,iou5Bs,lyGbBy,t7hBe,F8AAC,9TB.,OSnCe,m10L8, 11, | 0 oie aalmiqpuloetssowferseerawterraectreedmionvdeudpfrloimcetaech,sampleandextractwedwiithanon-pairingreagent; alloftho ! pressure lTihqueisdechrroemxtartaoctgsrwaephrye<alneaclryozsepdrqauyamntaistastsipveelcytfroormpeetfriyv(oHrPoLoCct-aEnSeMsSul)f.onAantael(yPFiOdSe)nbtiyfihcaitgihon | `mwaatsevreirailf.ieSdabmypcloemspwaerriseoqunaonftimtoalteicvuellyaerviaolnua-tPeLdaCgraeitnesnttiaosnpetciimaelolyfptrheepeaxrterda,cSiveed-apnoailnytteexatnrdacstteadncdaarrvde. wi(t2h.i=n0.1909%6)o.fMeatxrpixescpvkatelcueaelsdi.bration check,analyzedinthe middleofthe analysissequence,were | 'ESMSMTS.hTehpirsetseecnhcneoiqfuPeRprOoSviidnesseaveraadldoifttohneaslameplgeos (of1Acterhtraoinutgyht6oBt)hweaansavleyrtiefiieddeuntsiifnigcaHtiPoLnbCy-. | sppecirficifalolnymm(ownaiizto=rr4i9ny9g)f.ragmeatsdaughte ous (m/z-80,99,130,180, 230)characteristicofthePFOS | previousl`yT.hSepPeFcOifSivcarleusuelstisantrheaitstbaicnhderdetportodiuscirbeiploirtty.analysisarewithin 11% ofvaluey scollected Experimental summary: | SamplepIrnepaapraHticoonn:trJoolne-pdaeinrvinigreonxmternatc,toannfoopaiingreagent etrabuty:lammoniumsulfate(TBA) is ] (`PusOeAdAto)e,xatrreasceltehcetainvaellyytteafrrgeotmedtbhyemtahterciaxt.iAonniicorneiacgecaotm.pSouubnsdesq,ueknet tPoFtOheSafonrdmapteirofnluoofrtohoectTaBnAo-satneion. p`airmtheaefnaolrtyHtPehLCt-raaEnSsMfneSrarneoadltyolsainso.n-polarorganicsolvent (ethylacetate),rod,andreconstitutedin I HPLC:ChrstrtatetreseoertPoROnS | + BasedonItnhHePafLfCin,iatynoaflithqeuoatnoafltyhteefeoxrttrhaectstiaitinojneacrtye-dpahnasdepianstsheedtchorlouumgnhrealcahtirvoemattotghralpihqiucicdomlobuimlne.phasepassingthroughthcolumns,the analyteis retainedfo a characeisicamountoftime. For 'ePxFaOmSpsloel,uitnioanstaannddtahredasnoalltyitoene,xPtFraOctSemd faryoemlsteeraaitn1t0h.i5miannaultyessi.swReetreentmiaoincthiemdefsobweittwheienn1a%sotanntdhaerd. | HPLCsystem. | 004403 oss 1 || ESAS:ADnaleysitsoaefndPFmcoOnSitsttoarinidnagroodsfintnhdeimcoatleesctuhlaartihonprimaryloncharacteristicofPFOSism/z =499 {`a0mmua,xciomrirezsepsoenndsiitnigvtiotyt.heAmsacsasnoofftmhe/za=ni1o0n0itcosu1r2f1a0c(tanbt(gCaF tSOi)w.vaTsehaiclssiaocnollweaycstme)odn.itoredselectively ESMSMS`:SevCeraolsnampflesiionfrthnisamsteetaiwdeternetiiafniaoclaytznieondbyESMSMStoverify theideattyoftheFOSanalyte coomnp.EouSn-dMiSdMenStiifsivcaetriyosni.mAilsarintoEESSMMSS,,aexccocmppttohuandtisptaecdidfsiacnoanddistsieolneacltdeidm.eAnfstieorsn eoflceetacitnthtyeitseoolenct,ed , ofrnagimecnhtasr,accthearriazceterifsutribcoerfbhyesmmolaeschuilneg,aitraepcarretawtietdhahnidgdhectneccrtgeyd.gas.As resultofthesmashinongi,c i ion smaFsohreedxaatmopotlhefeo,rroPnFssOScahna2l5ysi30s,aimoun 4(9c9oirresseploencdtiendgastotSh0e5c)o,m9po9uanmduspe(ccifoicrprreimsarpyootonnd.FSiTOnhi)gs, 130amu (corresptoCnFdSiO5n)g,180am (CoFand.23S0amOu(5CyF)SO,y). I(PisFprOeseSnt 1 inthesamples, cachofthes secondaryfragments sdetectedat thedetector. i QualityEcaoncthroslesrua msammaprleyw?as extracted induplicate; standarddeviation valuesareincluded n thetable | ++ ot Seasmptless.;reTcwovoemriydf-olrevtehle2i5xpmpabtrainxdp4i5kepapbn)alquyasliistwyecroendterotlesrmaimnpeldefwoebreewaintahliynze1d0b%eotfweeexnpeevcetredyvfailvuees. | p`rBee-ccaxutsreachtuemda,nsspeikreadwwiitthhouasttPaFnOdSarisdcnoontcaevnatirlaatbiloen,osfamPpFlOesS,oafpnodorlee-desxetraacptuedr.cThhasiesdpfrroem-Seitgmraawsceetrreeid uannddearfstteorotdhtesoabmepvleersy.siQmuialnatrittaottihoensoafmtphleesmaamtprliexs.iTshbiassefivoenpotihnetleixnteraacrteegdrceusrsvieonweaqusaatniaolnydzeerdibveefdofrreom 1 tahpepaevnedreadgereosfultthseatblweo.bracketing curves (r*2 = 0.996).SpecificQC parametersareavailable on the . | L~iofQnAusanttihietsaettsiaomnpleswerepre-screened, itwasdeterminedtha optimizinginstrumentaldetection. | Liqmuitaswnatsiotwtaansct1ce0ispsoparnby.fAorlalcscaumrpalteesawnaelryesiqsuaonfttihtastee"dmwiedl-loavbeolv"etshaimsplleevse.lA. saFetuellimi,tof | ion ae | HInPsLtrCumsenetaml specifics: : `HewlettP`acCkoarldSuemriae:s K11e0By0eLtsiaqtsuoidnCCieharomatograph. | 2S xum1p0a0rmtmicslizee FSloolvwerntteA:: 32.000mlMLiAmminm.oniumAcetate ] SoSloviveenattGBr:adicat:Methanol | `4Hold52.9%10B%Bi0nf9o.r9503m.5i00nmsi.ss. HRoeltudm24to54%5%BfBoirn31..5m5i0mnisn.s. Injection volume:10 iL Electmrasossspepcrromaetyer | `MicromasMsPalatsforsm2IL.T4SAyoPfItnMwaaxrsesSpectrometer,"ChickTM 004404 oss 2 i Conevoltages: 60v SM`ooAundraelcy:ezeetsleevmcpaterrooamstpumrrpaery:nesesguarteisv:1e105.0C0.0043mBar, 0.000079mB ETloencstr:o4d99e,:4c13o,ss36l5ow ElectrosprayMSMS. ; MicromasC`soQnMueaavtotsrlogiIe3I.s1LA:SPoSyfI0tMvnwaaserseSpectrometer,"MadelineTM | Collgasiensergiy:o45nV/ Mode: electrospraynegative 1 ``SAonuarlcyezteesmvpaecruautmurper:essures1:105.0C0.0043mBar,0.000079mBar . "Primlona:4r99y, Dau ionsg :80,h 99,1t 30,1e 80,r 230 || osm 004405 3 | Fluorine Analytical Chemistry Team 7 Contac: sous SSaammpplleessERxetcreaicvteedd:: Samples Analyzed: aannsiss 4/164122/98 AsCalyasislby:CiurbveRranga1E0tSppMbiSto9o6pnpb 00% loofpCealibrationCurve: 0.99 ~~ * 00: 10pm PHASE IX, part 2: Blind repeatability experiment } | Ee [Twos| Pos faa] 2 1 1BAs 2[8 o+/e-0s.3| 2a 8 a2c | o[srsveoo2s|| uwsi 40 e3sa || essarr2| 54 @ | ures | a sa | mens |e fouls 9MoBs.c 3|4aw+s/]-|1 3w0 B1om || aseirea f| aoew 25 || moersa]| naaa 1 | wera) ae n [0+] ] QC Results | [c[uQiCbCmrteoatnecook, 24.9pp [Catibration Check, 49 ppb | [limes [Calibration Check, 24.9 ppb |Calibration Check, 24.9 ppb | |CalibrationCheck, 49 ppb 2 We R5ec6o%very 10% 2% 0% 98% : P+ 3 : . | | 21--1L0OQD == LLiiooffmDmQeutuieaicttitiaottnion | s3MsEsaviroomental Lab 004406 PHASpEaItX2 3KrMisEHannvseinr-oSrn.mAennatallyLCachebmaoirsltatory-Fluorine AnalyticalChemistry Team FBluuiolrdiinnegA2ua3l5y-t0i9calChemistyTeam 127786018 Preliminaryresults: Analysis of POAAin humanserasamples Summary: . g `Todate,datafrom nine phasesof a studyof fluorochemicalsin humanserahasbee collected. i PPHHAASSEEII::SeQmuia-aqtuaanttiivteaatnivaelyisnivseostfiPgFatOiSonaonfdPPFOOASiAnisnasmapmlpesleosfopfoploeodlheudhmuamnasneUraS serasamples,1997-1998 1 } #4 nnn PHASEIII: Quantitativeanalysisof PFOSandPOAAinsamplesofindividual's sera, 1998 - PPPHHHAAASSSEEEIVVV:I::QQQuuuaaannntttiiitttaaatttiiivvveeeaaannnaaalllyyysssiiisssooofffPPPFFFOOOSSSIiinnnhihssiatsmotporrliicecasaollUfUSsSessreaerrfaarssoaammmippnlldeeissv,,i1d91u69a07ls0.sin-8r0usral. `northernChina, 1985-1997 | PHASEVII:Geographicaldistribution: aquantitativeanalysisofPFOSinseracollected. d"aaPrtHoeuAnSndoEttVhsIeupIUpn:liiQteuedadnt0SitEtanattevisiv,era1on99an8lmyesinstoLafbPlOSinhistoricalsamplesfrom Sweden,specific 1 PHASEIX,part1:QuantitativeanlyssofPFOS historicalUSsamples,1948-1951 "EPxHpAeSrEimIeXn,tpart2: PFOSlevelsinindividual'shuman serasamples;Blind Repeatability | `Theanalysisofperfluorooctanesulfonate (PFOS)hasbeentheprimaryfocusinthesephases; quantitativedata for perfluorooctonoate(POAA)has also been collected for several phasesofthe study. | Dp`uaarnetaltoyosfitts.ihim`sTeohceonnfsgtoroaisilnttnusl,dgrwye.oephRoarewtpvdoeeirsntcosnrtdidebegtevasoiotlbiesndegttrhhveaetsPiaFomnOesSoataftneaPnlOtyiAsoAinsltieovttelhdseiPanbOsoAovAemhaeasovfwetbeheheeasnvaemiptslouetoshdaetnoPaJFleyOlzS.e.das | `Mandelinthe3MMedical Department. LOQfor POAAis 10ppb. 1 `NNuommbbeerrooffssapmipleesswwiitthhPPOOAAAA >>LLOOQD ==04 . J Comments: `Totalnum ofsab mple es=r 12 ] |. NuTomtableurmobfesraompflsesawmitphl=Pe3Os1AA >LOQ=4 Comments:Datahasnotbeen examinedcarefully : ] Number ofsampleswithPOAA > LOQ = 0 `CToomtuamln eatsu :Dam oaf shb aampnle oetsb=r e1n0precisely quantitated | 004407 ounuss 1 NNuumzbbeerrooff ssunnpplleesswwiitthhPPOOAAAA >>LLOODQ ==06 i `TCoomlmeunmtbs:eDraotfashaamspnloets b=ee6nprecisely quantitated - | `nNourmthbeerrnofGsianmgpleswithPOAA >LOQ =0 TNoutamlbnerouf suommfplsebasmwpeiltehsP=rO9AA >LOD = 3 | `Comments:Datahasnotbeen preciselyquantitated 1 fee | ] . SicoslfrlousndehecwUintltAdeSAitdes10021 2008, 1 250) `TNoutmablenruomfbaermopflseasmpiltehsP=5O5AA >LOD =o20 e `Comments: Datahasnotbenexaminedcarefully PHASEVI:QuantitativeanalysisofPROSInistorica samples fromSweden. NTNuoumtmabbleesrruoomffessrauommppfllseessawwmiittphhl=PPeOO2sAA0AA >>LLOODQ == 00 `Comments: Datahasnot beenprecisely quantitated | `NTuotnableurofmsamopfblsemswpeiltehs=rPO20AA >LOQ= 0 Commeots: Datahas not beenpreciselyquantitated | Experimentalsummary: ;| | SamplepIrenp.arpalticoonn:tlroolnlpeadieraivnigreoxntmreaactt,ieonn on-paiingreageat, od 0extracttheanalytefromthematrix.Anioniccompounds, : itkeerPaRbOylSaamnmdopneriluomosruolofcittaen(cTeBtAe), is | (pPaOiA,tAh)e,anaarleysteeleicsttirvealnystfaerrgeedtteodbaynothn-cpaotliaornoircgraenaigcesnotl.vSeunbts(eeqtuhyelnatceottahteef,orrimeadt,aionndorfetchoensTtBiAtu-taezdinon - `metfohrMaSannaloyslis I PE -- i | HPLC:CIhanrEaPctLeCri,satniacrleitqeunottioofntthiemeexsftorracPiOsnAeActedandpasethrough chromatographiccolumn. i | Bphaasseedpoanstshienaftfhirnoiutygohftthheecaonlaolmyat,etfhoetahneasltyatteiiosnarreyt-apihnaesdfe oi t8hccholaumraraelcataeimvroetuointtshoeftHtiiiqnucei,dmFoorbll`example,ina standardsolution, POAAmayclueat 10.5minutes. Retetion timesbetweea astandard PHOPALCAssoylsutteimo.nandtheanalyteextractedfromserinthisanalysiswerematchedtowithin 1%oathe. | taomum,axciomrAirnezaselpsyoennssdiiistnoigvftitoPytO.AhFAmraasgtsmaennodtfaatrtdihsoannndoiifoctnahieecsstPuhrOfaAaAtcthaaenpnitr(oiCnmFacriaynCibOoezni)cnh.daurTcahecitdes;roiosnntiw3ca6o5sf,mPoOhnaiArtAaoircseimditlseizlceo=c4fti1v3ely | ose 004408 |) dSeccaanrboofxmy/lazt=i1o0n0pr0o1du2c1t0,r(aeesgualtti.vBeoontlhyi)ownasswealrs:omcoonliletcotreedd.inmostof thestudyphaseslistedabove. A ESMSMS: Confirmationofanalyte identification | ion. ES-`MSeSvMeSrailssvaemprlyessiiminltahritssoeEtSwMeSr,eaenxacleyptztehdabtyiEt aSdMdSsaMnSatddoivteiroinfaylthdiemiedaesaitoitny ooffctehrteaPiFntOyStaonalyte. | icoonmpisocuhnardaicdtenetriifziecdaftuirotn.heArsbiynsmEaSsMhSi,ng2cioampaprtawintshpheicgifhiecnoerngiyssgaesl.ecAtesd.8Aefsueltsofethecstmtaihssohelinencotn,geid,c i fragmentFs,ocrhearxaactmerpisltefioc,orfPtFhOeSmoalneacluyslies,,arieocnr4e9a9tiesdsaenldecdteetdeactetdh.ecompoundspecificprimary on. This _ | io13n0iassmmuas(choerdrienstpooontdhinegr toonCsFs:uSc0h5)a,s 81080amamuu((coCr,rFe.sSpOoyn)d,inagntdo2S3O05)a,m9u9(aCmFSuO(5c)o.rreIsfpoPnFdOiSngistopFreSsOca)t, inthesamples,cachofthesesecondaryfragmentsisdetected at thedetector. 3 1 QwuaaslintoytpWcoohsnsetibrnloepl,ossausmbslmaeam,rpslyee:rssszaemapnlde/sowretriemeextwrearcteeldiimnitdeudp.lMiceattheaannodlanballanykzsedwierneduapnlailcyazteed,pOefrieonditchailsly i tthorcoausguhreecacohmapnlaelytiiscoallatsieoqnueonfcteh,estypaicpall.yMeivdelreyvfievleqsuaamlpilteysc,otnotrmoolnsiatmoprilnesswtreurmeeanntasltaybzileidtpye.rBieodciacuaslley - ehxturmaactnesde,rspwiiktehdowuittFhFaOsStiasndnaortdacvoaniclebnltera,tsiaomnpolfesFFofOpSo,oalneddrsoe-reaxpturracchtaesde.dThriosmprSieg-meaxwterraespcertreae-ids : || fuonrdmerossttopohdas0beseovfetrhyssismtiuldayr,PtoOthAeAswaampslaelsmastpriikxe.1dannadlelcxatrsaecstePdFtOoSwoarsmevasatlaunadtaerdvdceursruvs.aInnetxthreasceted; | cathseetsw,oqcuuarnvtietsatbiroancokefttinhgetshaempslaemsplisebsa.sedom thelinearregression equation derived romtheaverageof w_ Limitof"QTuhaenctuirtarteinotnanalyticalmethodhasbeenusedtoevaluate alargerangeGf PFOSlevels - extracd, | f`sraommpsleesrw;itthhePmFeOtShcodoi nnot copteiminiznetdtfhoerrrparanegcetisoefil2o0wo--l6e0nvpelps..(<A5psppebc)iqaula,ntoiwt-alteivoenl,cbuurtvienhsatseabdefeonr :: | Gpbrseepravreeddifnornporne-cpilsaenqtuawnotrikteatrisveereaavarleuvaetriyolnoofw,PiFnOfSulteuvreelasbnaeltywsese,nP1-O1A0Appwbil.lAbse tevhaeluPaOtAedAalgeavieolssta special o`wSlamepvleelscduersvieg,nsaitmeidl<arLoOQthcotnptraeipnareesdsftohaPnF1O0S.ppb POAA.Iffurtheraccaundrpreaciscionyin | vqeuarnstuistaatsipoencoifalthleosw-elreveslamqpualnetsit(astaimvpelceusrlvees. than 10pp) i required,sapswillbere-evaluated cs ] Liofm"DTehtieecltitimoinofdetectionfo thisanalyticalmethodisbasedonanalyst recogaitionofthedistinct paenaalkyszheadpbeyretshuelctoinndgiftrioomnsrPeOpAorAtaendahlyesri,ss.hPoOwAneAasrtbaansdelairndemraetseorliuatilonnodfP1ObAraAnecxhterdaactnedd 1frloimnesaerrPaOAA | iUsnpolmaiekrer,.tP"iFThOceaSaut,pllaeonxwadalrtefvhelealcyrsa.htipoPeaoOkfitAhn,etbwhrheaenHcnPhpLerCed-sptceraaatk,cesotoectcchuaerlstiaonbteeaarpsprpeeecasiekfniicts,aDrtoespturncoehdcluecosiswballreeivlreyelctsoentnshtiaisottnaectncitum,rea.te A: | Characterizationofthesmaller peak(the branchedisomes)isverydifficult. * ' Qualific"aTthiiosndsa:tawillbeconsideredpreliminary nil th analyteextractedfromthesesamplescanbe | vcelreiafrileydiadesnPiOfyAtAheuasnianlgytLeCr-esMpSonMsSet.eTchhneimqueetshaondduusnetdifloritnshtraunmaelnytsailssoafstihteisvietyscaamnpbleesohpatsibmiezeendto more i vsatliydabtaesdbnoytLbCe-eMnpSeMrSfoprrmeveido.usly,butconfirmation of POAAforspecificsamples nthiscightphase 3 | i ounuss 004409 L+ 3% | Jastramental specifics: H`PHeLwCetstysPatcekmardSeries 1100LiquidChromatogragh l Column:Keysioe B2exta1s0l0Cmism | FSloovwennttsA:: 32500u0mmpMamAnimnim.coslniiezuemActaie SoSlovlveennttBG:radient:Methanol ] F45o1l0d.92109%0B4iBnf9o.r503.m5i0nmsi.ns. 1 FRoeltudmatt4o54%58%48fiorn3.15.m5i0nmsi.n. IInjnecjtieoncvt/olsiuamomen:ps.:102uL Tw or `E Micromc assPlt amtafsosrmso pIeTcAtPrIor mMeatesrsSpectromeer,"Chick" | ``CoMnevaolsta2sge4sLS:oy-f2t0nwv,axr6e0 SMooudrec:eetleemcpterroastpurraey:negativ1e15C. - 1. `TAonnasl:yz3e6r9v,a4c1u3u;4m9p9r,es4s2u1res: 0.000043mBar, 0.000079mBar Electrode!crosslow | E`MlieccrroomsapsrsaQyuMatStrSo ITAPTMassSpectrometer,"Madeline" | ``MCaosneeLvoylntxa3g.e1:So2f0wvare -- Colisiongas nergy:45 V LE | SMooudrec:e etleemcpterroastpurrayen:ega tiv1e13C. 33 | P`rAinamlayrzyeTrovma:cu4u1m3presares:0.000043mBar, 0.000079mBar `Daiuonsg:11h9,i16e9,3r69 ] Eleciode: Z-spray : . : ` : Lod . | ! on 004416 oa I