Document 4JpzmEBOa2RkJgYDD50bXKpJR
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ARRLG_ 1264 DuDPuPoonnttEEMMSSEEReRpeoporrttNNoo..1155:-0033
Study Tide
ACCELERATED BIODEGRADATION OF 8-2 TELOMER B ALCOHOL -- A PRELIMINARY SCREENING STUDY
Test Guideline `BTihoidsesgtruaddyabuisleistyth(e19c9o2n)ceptof OECD Guideline for TestingofChemicals; Section 3: Inherent Author Ning Wang, Ph.D. Study Completion Date 20 March 2003 Test Facilites CELeLntdraulPRoenstedarecNhe&moDuervselaonpdmCeonmtpany `ECnovriproornatmeenCteanltaendfoMriEcnrgoibnieoelroignigcaRleSsceiaerncches & Engineering. NGelwaasrgko,wDBuEil1d9i7n1g4-30601,01P,.OU.SBAox 6101 and HEa.sLkdelulPLoanbtodraetNoermyoourrsHeaanldthCoanmdpaEnnvyironmental Sciences Newark, DE 19714, USA Submitter DEuLPodnutPCohnetmdiecaNlemSooluurtsioannsdECntoemrpprainsey Wilmingion, DE 19898, USA EMSE Study/Project Number 1503/4842 Report Number EMSER 15:03
E eEMSeIESREIR1ee 155--00e3/4e8542------eeeeseeeoreNTeAIeNseNemeOnsPsPeaeTm ean
000033
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DvDuPPoomnEtMESMESREeRpeoprorttNNoo..1155:-0033
PAGE RESERVED FOR SPECIFIC COUNTRY REQUIREMENTS
EMSERI5-03/4842
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- OuPoMEMSERgoDm uPonN t EMSoE Reiports No. 1 5-03
CERTIFICATION OF AUTHENTICITY
ACCELERATED BIODEGRADATION OF 8-2 TELOMER B ALCOHOL -- A
PRELIMINARY SCREENING STUDY
Wsuep,ertvhiesiuonnd,erasnidgntehda,tdheicslraerpeortthaptrtohveidweosrakndcesucrraibteed rinethcisoorferptohdrt pwraoscepdeurrfeosramnedd urensduelrtsour Report by:
-- PTiRr thal 03/30/03
Senor Research Biologist Approved by:
aTr.chGaMnannoanT g,eFrAD. Horr
Bu0e3/30/03
SLteuhdiylyI2ni0t0i2ation Date: D2a0tMeaSrtcuhd2y00C3ompleted: SDEuuLbPmodinutttPCeorhn:temdiecNaemSoouurtsonnsdEnCtoemrpprainsey Wilningion, DE 19898, USA
SRS
mmo 000035
DuPont EMSE Report No. 15-03
TABLE OF CONTENTS CPOaTgHeEIRCeAsteirovneOdffAorESPCECi.fi.c C.OUNTY REQUITETIENS......crecceosvsssrcnnrnnnnrrns 2 TIE OF COMEIS.vrtsssnessnnsescscsetod
2.0 3.0
GMEOnEErriaallsStGU1dYMIEtOhMOUNS.AIr ON...e .vene rrr n r tn es r ]
3.2 3.3
TSeasmtpCleOMEUXICT.A.CHcOeNr8a0ArRrBssImYssSsIsSss.s.m.msvsveessvnrsmrsrnrssnnissnsesnsnionnsnnninneseccc012.
33.1" 332
ASnaamlpyltiecCaollMleectthioodnsafnodrETXesUtTSauCbtstOaNnce andr PIOGUo CIS..v ..cocee ceocer cecrcs rernr
12 13
4.0 ReSults and DISCUSSION...
A
SB CTO wommmmmsmt----
mm
--
6Fi0gureRe1fer8e-n2ceT.e..lomeBr Alcoh-- ol (8-2-- TBA)-- concentration ds urinr g then28-dray ts est.-- ....._ .....s ...19
FTaibgluer21:e EsFtliumoartiedde cCooncenntcraetinodnutsroirfngaftluhtoer2ii8na-otdeandyactieds.met.a.bo.li.t.esca.tvdavyv0r(e1r9-nJuclr- onn 20
`Tabl2e02:02D)aialnydtdemapyer2a8tu(re1re6ad-ingAs Uin Gthe-la2b w0he0re2th)e .tes.t w.asccocndvuctredr. Tmhie nns 21
temperature was recorded by a calibrated Dickson Recorder (Model THDx, serial#
ADPEDdiX A...
A --------
Table(A2-lA.ug-A2na0l0y2t)i,caalndredsualyt2so8f(81-26T-BAAUcGon-ce2n0tr0at2io)n.at.dvayr0r(r19r-nJsul-2002), day 14 24
a
Table Ack (SOME).
nis
26
`Table(AT-r2e.atmAennalty3t,icaadlaprteseudltbsacotferSipailkceulrteucroeveprlyusogfr8o-w2tTh BmAedfiruommitnhecosaatmepdlgelamsastrsiexrum
`TableboAt-3t.les)Anaatdlyatyic0al(r1e9s-uJlutls-2o0f0f2l)u,ordiadye c1o4n(c2en-tAruagt-i2o0n02a)t,daaynd0da(1y9-2J8ul(-1260-02A)U,g-d2a0y021)4..(.2.-.....27
TableAAU-g3 -(2C0O02Ra)nUd,IdUaEy2D8)(.1..6r-vArUrGn -2s00s2).....vwmrsonmr--en--_28
TableaAc-i4d.(2A-nPalFyOtEiAca)lartedsualyts0of(1f9l-uJourli-n2a0t0e2d),acainddmdeatayb2ol8i(t1e62--Apuegr-f2l0u0o2r)o.o.c.t.y.l ethanoicmoni
TabledAe-cSe.noiAcnaalyctii(dca2lHr-esHuDltFs-o2f-fDluAo)riantadtaeyd a0ci(1d9m-eJtualb-o2l0i02t)e,2aHn-dhedxaayde2c8af(l1u6o-rAou-g2--2002).......31
`Table A-6. Analytical (PFOA)atday 0
results of fluorinated acid (19-Jul-2002), andday 28
(m1et6a-bAoulgi-t2e0p0e2r)fl.u.o..r. ooctanoic
acid smn
Table(AP-F7H.A)Aantaldyatiyca0l (r1e9s-uJlutsl-o2f0f0l2u)o,riannadteddaayc2i8d (m1e6ta-bAoulgi-t2e0p0e2r)fluorohexanoicmacidms)
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DuPont EMSE Report No. 15-03
Table A-8. Figure Al
Componofebanctetrisal growth A fluorideStandard Calibration
medium
used for he est... CUIVE...........c.ccevcvrneerenen oo
34. 35
FigurTe A-A2AGcalciboramtmiosn curve used for 8-- -2 TB-- A quantifit cationofm samplesn TA 50-t 1 to
Figure A-3 Figure A-4
A A
chromatographof sample calibration curve used for
TAS0-3 usedfor 8-2 TBA 8-2 TBA quantification of
analysis samples
..................37 TA 51-1 to
FigureTA-AS SAcI8h0rdo-TmaA1tSo2g74ra1p1h0ToAfSsZam1pTle TAc 52-3 useo dfor 8-2n TBA anals ysis......e .............3389
FiguremiAx-6maCdheroinmastaomgprlaepmhastroifxsaTmApSl0e-s15T,ASs2am-p3leanmdatTrAiSx2T-A6,S23-016u,gaL"nodafmsettahnadnarodl acid
solventblank .........
A --------
Figure`mAix-6maCdheroinmastaomgprlaepmhastroifxsTaAmSpl0e-s15T,ASs2am-p3leanmdatTrAi5x2T-A6,S23-016u,g aLn"dofasmteatnhdaanrodl acid
FigureSOAI-Ve7ntAblGanCk/M=SCcOhroNmaItoFNgIrOUaMphPEoAfGdmEat42e.rial cr haracterizatr ionof 8-2 Tr BA test ]
SUBSIANGEUSCA I thiS UY APPENiX Be
red --------------
TablePBF-O1.EAA)naaltytdiacyal0r(e1s7u-lStseopf-2s0p0i2k)e,raencdovdearyyo2f(21-9p-e5r6fpl-u2o0r0oo2c)t.yloretchsanoic acid (2-
`Table2BH--2H.DAFn-a2ly-tDiAc)alartdesauylts0o(f1s7p-iSkeep-r2e0c0o2v)e,ryaonfdd2aHy-h2ex(a1d9e-cSaefpl-u2o0ro0-22)-dcecrenoeic aecidr( 4S
Table0(B-137.-SAenpa-l2y0t0i2c)al, arensddulasoyf2s(p1i9k-e5repc-o2v0e0r2y)o.f..p.er.fclcu.ocrvoco.ctanoic acsid (i PFOAm) atndayit
Table0B-41. 7Ana-lyt8ica6larne9dsud-lasoy22fs(p01i9k0-e5r92e-c2o)0ve0r2,y)o.fperfluorohexanoic acid (PFHA) at day 1
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DuPont ESE Report No. 15-03
ACCELERATED BIODEGRADATION OF 8-2 TELOMER B ALCOHO--L A PRELIMINARY SCREENING STUDY Author Ning Wang, PhD. 10 Summary
Rationaleof theStudy h"Tehsestetstsugbesntearnactee.s eWnhveirnoanmcehnetmailcaflaeenitnefrosrtmhaetieonnviprotoennmteinatl,lybrieoldeevgarnatdattoitohneipserosniesotfentcheeof 8m-a2joTrelrooumteers BthaAtldceotheorlm(i8n:e2sTtBheAe,nvCiArSonHme6n7t8a-l39f-a7e)ofmatyh bechselmoicwa,lb.ecTahuseeboinoldyeg2radation of tHhyedr8opcearrfblounosroinfatthedecmaorlbeocnuslearmeaeyxpbeectreedadfilybeacdcieffsiicbullteoforbemimcertoabbioallimzeetdabbyolism, therst of cmhiecmrioocraglamniasymsa.dapHtypoothheatviecaahlilyg,hemircmreotoarbgoalniicscmaspathcaittyhatvoetrbaenesnfoprrme-aenxdpopsoteedn0tiatlhley test cduelftluuroeriunnadeer8-f2avTorBaAbleungdreorwtfhavcoornadbilteigonrsomwalycognidvietiaonlse.aAertiensdtiwciatthiopnroef-tahdeapted bacterial cboinodtirtainosnfsorfmoartpiootnenptoitaelntmiiaclrfoobriaalgmievtenabtoesltiscmheomfic8a2l.TBSuAc.h Fotresetxmaampylepr,ofviadechoepmtiicmaall i not imnethaeboelnivzierdonumnednetr.suTchheofpetsitmsiyzsetdecmonisdiatlisoonuss,eifutli lersapliidkellyyitdoenbteifmyeptoatbeonltiiazled/biodegraded ibdieonrtainfiscfaotrimoantainodnqpuraondtuicftiscaatinodnhoifstrwailnlsfeonrambalteiounsporfodauutchtenitnitchsetsaunbdsaerqduseontfdaceifliintiattieve biodegradability sudics. Test System: "mTehdeipurmimpalruysbbiaocttrearinaslfocrumlattuiroenhpaottehnatsiaplroef-tghreowtnetins0ub-s1t0a0ncemg8/-2LTofB8A.2inTbBacAtewraiasl growth "dTehteertmeistnesdy.stTemhecobnascitsetreiadolfcuilntduirveidouraiglilnyatcerdimrpoemd taenstivndeusssterilal(lwaassstesewrautemrbtorueeast)m.entThfeuc(lietty. w`waass icnotnrdoudcutceeddaitntrootohmetbeamctpeerriaatlugrreo(w~t2h4mCe)d.iTuhme(8T-a2blTeBAA-8s)twoictkhsotlhuetbiaocnte(rmiaaldeinionceutlhaunmola)nd s`waamspkleepbtoitntlcelsowseedrebsoarcreifiinctehfeodraerxktraatctriooonmatnedmpaenarlaytsuirse.ofPtehreiotdeisctaclhleym(idcaayls, f0lu1o4r,inaantded28), acid metabolites, and loride (Fion). Findings: d`Tahye2p8r)icmoamrypabrioetdrwainstfhotrahmbatiiootniocf8c-o2ntTolB.A wSiagsnisfpiciadnt(>de7f0lu%oraitndaatyion14ofan8d-2neTaBrA10wa0s%. at aobnsdewrevreed idduernitnigfitehdetthersto.ugFhoutranodefhmemapostsenstpieacltbriotmreatnrsyfaonramlaytsiisonanprdocdoumctpsawriesroenmwointihtored for CanAalSyHti2ca7l65s4t-a3n1d-a5r)d.s:21))22H--pheerxfaldueocraofolcutoyrtoh-2a-ndoeiccenaociidc (ac2i-dP(FO2EHA-,HDFF(-C2F.:D)A,,CH:COOH, F(CFCF-CHCOOH, CAS# 161094-76-4)3)perfluoroocianoic acid (PFOA,
EMSERI5-05/4842
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_ DuDPuPoonnttEEMMSSEEReReppoorrtNNoo..1155:-0033 `FC(ACSFH#3C07O-O2H44,).CAOSt#he3r3p5o-t6e7n-t1i)a;l atrnan4ds)fopremraftliuoonrophroedxuacntosicathcaitdfo(rPmFeHdA,duFri(nCgFth:eCtOesOtHw,ere. not determined.
Conclusions: Uotnhdeerruntihdeetnetsitfcioenddtirtiaonnssf,or8m-a2tiToBn Apriosdbuecitn.g rAalptihdoluygthraPnFsOfoArmiesdotnoeoffltuohreinaitdeedntaicfiiedds and mmeettaabboolliitzees,d ittoafcocromunPtFedHAfora<nd2m%oayfmnaotssbebaalnanuclet.imaPteFO(sAtaablpep)armeenttalbyolmiateyubnedefurrtthheertest conditions.
20 GENERAL STUDY INFORMATION
Study Objectives Dectoencremnitrnaetitohnecbhiaontgreasnsdformuatiaobrniopdoietgerntanidaaltoigfon8-e2sTtBwiAthbyopmtoinmiitzoerdincgonidtistions Determine the degreeofdefluorinationof8.2 TBA Identify potential metabolitessuchas Muorinated acids durinthg test Assess whether perfluorinated acids can be further metabolized Determine possible biodegradation pathwaysof 8-2 TBA Co1n0fpoitrenmtieaxlpaebcitoattiicornetmhoavtamlasmsecbahlaannciesmwi-sllvboleatdiilfiftiycualntdtoadascohripetvioen.in cold studies due:
TestSystem Justification "The test system is modified from OECD 302 guidelines and was requested by the submitter.
Study Personnel
EL
du +
PCoennttdrealNReemsoeaurrcsha&ndDCeovemlpoapnmyent
-
Corporate
Cente
for
Engineering
Researc-h
HaEsnkveirlolnmLeanbtoarlataonrdyfMoircHreoabilotlhogaincdalEnSvciireoncnemsen&taElngSicnieeenrciensg.
Management:
JELo.hndTu. PGoanntndoen,NePmh.oDu.rs and Company
`CCeonrtproarlatReesCeeanrtceh afonrdEDnegvienleoerpimnegntResearch
GElnvaisrgoonwmeBnutialldianngd3M0i0c,rPo.bOi.olBoogixca6l10S1ciences & Engineering
Nanedwark, DE 19714-6101, USA
SH.asMkaelrlkLKaebnonreadtyo,ryPfhoDr.Health and Environmental Sciences
Newark, DE 19714, USA
EMSER15-03/4842
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DuPont EMSE Report No. 15-03
Study Director: Amslytcal Chemists: Technical Personnel:
Ning Wang, Ph.D. (ECedntrualPRoenstedaercNheamnoduDresvaenldopCmoemnptany CEonrvpiorroantmeenCteanltearndorMiEcnrgoibnieoelroignicgaRlesSceiaerncches & Engineering NGelwaasrgko,wDBuEild1i9n7g143-0601,0P1.,0.UBSoAx 6101 EBodgduanPSonztstdcekN,ePmhoDu.rs and Company NHeaswkaerlkl,LDabEorUaStoAry1f9o7r14H,eaUltShAand Environmental Sciences VEiLaddiumiPronCtapdkea,NePmh.oDu.rs and Company NHeaswkaerlkl,LDabEorUaStoAry19fo7r14H,eaUlSthAand Environmental Sciences KPeairtihckB.W.PriFcokletsto,mD,uDPuoPnotntHaCseknetlrlalLaRbeosreaatrocrhy& Development Richard F. Rossi, DuPont Haskell Laboratory
Study Execution Dates
Experimental Start Date: 19101-2002
Experimental Completion Date: 19-Sep-2002
Study Completion Date:
20March2003
30 MATERIALS AND METHODS
31 TestSystem 311 Test Substance
Name: Synonym: Active substance(s) CAS Name:
Molecular weight CAS Number(s):
82 Telomer B Alcohol (PerfluorooctyDethanol, 6.2 TBA 82 Telomer B Alcohol, 9% 313-D4e4c,a5n5cl,,6677,88.99,10,10,10heptadecafluoro. 464.12 g mole" 78307
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DuPont EMSE Report No. 15-03
Structure:
rF
on
.FF
F
FF
F
"F
F
3F
Lot Number: EMSE Sample Number: Concentration ofas.,nominal: Concentration of as. analyzed: Major impurity CertificateofAnalysis Date: Date Received: Solubilitya 25C: Vapor pressure: Stability: Appearance/Calor: Storage Conditions: Safety Precautions:
P.00/001 E03386.80 99% 99.2% 0.8%asCoF1sCF=CHCH;OH (Figure A) 13-Sept-2001 26:Mar2002 "140g? 0023 mm Hg Stable at ambient room temperature White solid Room temperature; keep tightly closed g`lWaesasrelsab coat, protective gloves, and safety
3.02 Reference Substance None
3.03 PreparationofBacterial Growth Medium a`uTthoec2la0v%edy.eaTstheexbtarcatcetriaanldgdriofwfetrhenmtemdiinuermalwasstopcrkespoalruetdiobnys dwielruteiopnreopfatrheed dainfdfewreenrtestock sstoelruitlieofnislt(esreeedTianbtoleNaAlgenfeor|deltiatierlsfiolferthuenimtesdainudmwacosmsptoonreednas).roTohmetgermpoewrtahtumreed.ium was
3.04 Test vessel Coating with 8-2 TBA solution wOante1r5-a1n4d-21000u2L, oGlfa8s-s2sTeBruAmsbtooctksso(lu1t2i0onmi(.3 vmogl/ummeL)iwnecrtehafniolll)edwwaisthin1j0e0ctmedLoinftsoteeraiclheof 3tdheaybosttaltesrofooma tfeimnpaelrcaotnucreentwriatthiaonboouft320500uRg/PL.M sThhaekibnog.tleTshweebroettsleeaslewdearndriinnsceudbwaittehd for sterile deionized water and capped for the test the next day.
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DuPont EMSE Report No. 15-03
3.15 Adapted bacterial culture preparation
Afapcpirliotxyiwmaastecloyll5e0ct0emd Lfroofmianedruisattriioanl twaansktseosnlu1d3g-eJufnreo-m2a00n2inadnudstwriearlewsaesnttetwoattheerttesrtealtabm.ent
U`Tphoenslaurdrgieviwnagsatmtihxeetdesbtylabbritehfelyssahmaekidnagy,thtehejusglutdogseuwsapsenadsstihgenmeidcarnooIrDgannuimsmbse.r EA9ft3e3r86-87.
stheetlsilnugdgtehewsalsudtgreanfsofrerarpepdrtooxiam2a5te0lmy L15stmeirinletocerllemcoulvteurceoafrlsaeskmtahtattercso,ntaabionuetd2abmoLuta5li0qumogt/oLf
orfo8o-m2tTemBpAeriantu1r0e0bmyLpebraicotdeirciaalllygrtorawntshfemrerdiniguamb.ouTth0e.5bamctLeroifalthceulctuulrteuwreasinmtaoianntoatihneerd5a0t mL.
p5o0l-y1p0r0ompygl/eLnoeftu8b-e2 tThBatAc.onTthaienceudlatubroeutwasmmLixoeftdhbeybcaocntsertiaanlt gsrhoawktinhgmaetdaibuomutp2lu1s0 RPM.
`The day before transferred into
tah2e5i0nimtiLatifolnasofktthheatteasltre(a1d8y-Jcuoln-t2a0i0n2e)d,
0.5 100
mLof mLof
the the
bbaacctteerriiaall
cgurlotuwrteh
was
`cumletduiruemwpalsusce6n0t0riufgug/eLdoafn8d-2thTe BpeAlleatndwaisncruebdaitsesdolovveedrninigthhte agrroowtohm mteemdpieruamtuarfet.er Tdihsecacredlling
tasheisnuopceurmlautmafnot.rth`eThtiesswt.asThenstmeLp woafstrheepewaatsehdeodncbaec.teTrhiael wcaulsthuerde wbaacstetrriaanlscfuelrrteudrewwiiltlahserve.
prleafsetrircedpitpoetatsektiollaed20bamctLerisaclinctuilltluatrieoannvdilwaiallndbewausseadutfoocrlaabvieodt.icTchonetraoultotcrleaavtemdenctu.lture was
3.1.6 Activated Sludge Collection
AMupnpircoixpiamlatWealsyt4e lTirteeastomfeancttiFvaacitleidtysl(uPdgOeTWwa)s-cAoelrlieacttieodnfBraosminth#e C2iotnyo1f6WSielpmtienmgbteorn20(0D2E.)
`wUapsonmiaxreidvibnygabtrtiehfelytesshtalkaibn,tghtehesjluugdgteo wsaussapsesnidgtnheedamincrIoDornguamnibsemrs.E9A3f3t8e6r-1se0t0l.iTnghethseludge
sludge sludge
for approximately 15 min to was filtered througah nylon
remove net with
coarse apore
matters, the upper aqueous sizeof 85 um. The filtrate
phaseofthe was inoculated
iinntooctuhleumbaictnearisaelpagrraotweteh mxepdieumraitnodmcoienncdnuubctattesdpiokveerrneicgohvte.ryTohfifsobuacrtesrpieaclifciucltfulrueorwianasteudseadciads:
standards, which representative of
wfleuroeriqnuaatnetdifaiceiddsinretchoevsetruydyf.roTmhtehseeasdpaipkteerdebcaotveerriiaelscaurlteuraes.sumed
to
be
also
3.7 Test Units
Test vessels crimp caps.
were 120 mL borosilicate glass serum The pre-cleaning was done by rinsing
bottles with pre-cleaned aluminum-lined the aluminum foil and septa with methanol
once and then with sterile deionized water three times.
32 Test Conduct tFhievealtyupmeisonufmexfpoielriamnednsteapltatrweiatthmemnettshawneorle ocnocneduacntdedt.heTnhweitphres-tcelrielaenidnegiownaiszeddownaetebry trhirnees:ing times.
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320
Treamen|t - 600 ug/L of8-2 TA `ICultunre ociunColateduBottmles
in
Bacterial
Growth
Medium
plus
the
Bacterial
Foofroaftthoetaglroofwt1h5 cmoeadteidugmla,ss0.s66e3rmuLm bofothte w(a5srheepdlibcaactteesrial3 csualmtpulrei,ngantdim&eApLooinfts)8,.229T:3A4mL.
bstooctkssowleurtieocnr(i3mmpegd/wmiLthinpreet-h<alnoela)newdearleuamdidneudmtfooielacahnodftPhTeFEgllassiscsoenrusmepboat.tles. The
322 Ineamem2- of8:2 TBinA BacterialGrowthMedium plus Kill
oFB fotrhaa etogtrac olwCotufhlt 9tmuecrdoe eaituiemndr ,Cgo0l.aas6i ts6e3sdema BrLoumuol lfbteohsetksil(l3edrebpalcitceartieasl cu3lseam,pl0i3ngmtLimoefp0oi2ntMs),N2a9C.N04,maLn.d b6ouulLoe.f 8T-h2eTbBoAtesstowcekrseolcurtiimopne(d3 wmigth/mpLrei-ncletahnaendola)luwemrienuamddfeodl taonedaPcThFoEftshieligcloanses sseeprtau.m This treatment serve s an abiotic contol
323 Treatmen3t ~ Growth
ls
i
lum ir
tles for
8oF:fo2trhTaeBtgoArtaSolwpotifhk9emceRadenitceuodmvgealrnasdys0s.e6r6u3mmLbootfetshe(3wraesphliecdatbeasct*er3iaslacmuplltiunreg wtiamseapdodiendt)h, 2e9a.c3h4ofmhL.e PgTlaFssEsseilriucmonbeotstelpe.a The bores were crimped with pre-<leaned ahuminum fol and 8At2cTacBhAsasmtopclkinsgoliutmionp(o3inmtgs/(mDLa.yi0n,e1t4h,anaonld) 2f8o,3ibnaoltcsoncweenrterastpiiokneodf(d0os0edK)EwLi.thT6heuL of sboatmlpelseweextrreacsthioank.en at 200-300 RPM for approximately 30 min before sample sllcion and
324 TFroreaattomtaelnoft49-cGorstoedtghlMasesdsieurummplbuosreBsac(t3erreipallicCautelstu*r3eIsnamopcluilngumtiimneCpooiantte)d,B2o9r.t3l4emsL golfatshsesgerrouwmthbotmteledsi.umThaendo0t.6e63wmeLreocfrtihmepweadswhietdh pbarcet-e<rlieaalnceudltaulreumwiasnaadfdoelda1n6dcach ofthe HPaTsFEbseinliacdosnoerbseedpao.tThhisgltarsesawtamlelntofwtahseutsesetdv{e0sperlosviwdaesaanvaiinldaibclaetifonowbhieottrhaenrst8o:r2maTtiBoAn.that
3.25 Ireamen5t - Growth Medium plus Bacterial Culture Inoculum innon-Coated
Boies
For a total of9 non-coated glass serum bottles (3 replicates x 3 sampling time points),
ac2a9nc.dh3Po4TfmFtLEhosefigtilchaosesngesreosrewputtmah.bmoeTtdhsii.sumtrTeahanetdmbe0on.tt66ew3silmweLsrereofvtcehraiesmswpaeamdsphwleiedthmbpaartctr.eircxilaceloannctuelodtluhrfeourwans iaddfeoidl to quantification of fluorinated cid metabolites by LOMS/MS.
326 SpOink1e7RSeecpotveemrbeyor f20F0l2u,oraisneapatreadteAcexipdesrifmreonmtSwaamspcloenMduacttreidxto determine the pike recovery ofifllReudowriitnhat3e0dmaLc.idosfbraoctmerbiaacltegrriaolwtchulmuerdisuammpplleusmtahtreibxa.cteEraicahl oifntohceulsuammphlaetbhodlbeeewnas pprree-pcalreeadntehd hduamyibneufomrefo(ilSeancdonPT3F.E1/.6s)iicFoonrsdsaeypt2asaanmdplweesr,etkeepbtotsthlaeskewnerate crimped with
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w~e2r5e0sRpPikMedatwiotoh mMutoreinamtepdeacrid0aamltilxuoswortlhueetiobnac(tmearidaeofrgormowi.ndiFvoirdudaalyst0oscakmspolleust,ionbiontes eptehrafnloulo)rtooocatfyilneatlhcaonnocienatcriadti(o2n-oPfFO2E0A0;pgC/ALSf#or2e7a8c5h4-a3c1i-d.5,TDhuePofonutr,ac2i)ds2Hu-sheedxaarde:ec1a)fl2u-oro-
2-decenoic acid (2H-HDF-2-DA; CAS# 161094-76-4, DuPont);3)perfluorooctanoic acid
((mCPaAFtSOiA#;3c0oC7ntA:r2So4l#-.43,T3h5+-e968b7o%-t1,i,lTe9sC7Iw%e,ArmeOeasrkhiawek)oe.ondAatPnro2ot0dh0ue-crt3s03)0;bRoatnPtdlMe4s)fwopreesrrfplpnuorotorxsoiphmietkxeeadnloy0ic3s0aecrmividen(abPeFsfHoarAme;ple sS3oaclmurptilimeopnecoidnlbleetochttainosonlwa)entrdoeasasfpmiinpkalleedceowxnitctrehancfttlirouanot.riionAnattoefdd2ac0yi02dgsmaiLmpxlfsoionlgucttaiicomhnes(pemoiadi.dnteA(f1rn9omSoeipnt3tdeibhvmiobedeutrarsl20s0t2o)c,k wToerrppnrootxsipmikaetdetyo3s0ermvienasbesfaomrpelseammpalteicoclolnetrcotli.on Tnhde sbaomtplseweextrreascthiaoknen at 200-300 RPM 327 TCesot nditanidSoampnlisng
3.27.1 S"TahmepclreimIpnecdugblaatsisosnerum botles wer incubated with 200-300 RPMofshaking at oom \hermopuegrhaotuutrethientchoeurdsaerok.ftThheesrtuodoy:m temperature ofthe st lb was monitored and recoded
3272 Sampling Interval 5 crimped serum bots rom Trearment and bots from the restofweatments were
sampled for extractionof 8-2 TBA, fluoride, and fluorinated acids at day 0 (19-Jul-2002), day
14 (2-Aug-2002), and day 28 (16-Aug-2002).
3.27.3 SAnasmyptliecaSltosarmapglees were stored at 10 to 20C.
33 SampleExtraacndtAnialoysnis 33.0 SCampolellecantdEixtroactnion
3.3.1.1 Sample Collection
wiAettheddatyrusawr0,nd1w4u,iptsahinadded218o0,w-cnmrL.impApoleytdoptsraaolmpopyf1llee0nbemostyLtrlieosnftgwheeaerentedrsetwmsodveiindujmefcrftoermdomtiheeoaschha1k5oefrmtLah.nbdottheebsotwtleerse polypropylene tube that contained 100 kL of SN sodium hycronide for fluoride extraction.
33.02 SFlaumoprlideeEexxttrraaccttiioon:n The 15 mL polypropylene tubes containing the 10 mL test medium plus N53a.3OHuLweore6fNincHuubSaOte,dwaatsoaodmdetdetmopecraacthuroeftfhoer 3ub-e4s towinth e250v- 3tt0h0etaRePtlMmoefdiishuzamk,inTgh.eThen sample tubes were sored at approximately ~20C for fluoride analysis 3f.lu2orTiBdAe aannaldyfsisl, 0u.3o4 mraLciiondf6exaNtrtaHcteSiOodn4:aAnfder30wmitLhdofrcahwiilnlgead 1M0TmBLEowfeterstimnejedciteudmifnotro the s(a2m5p0l-e3b00otRsE.M)AfatterroMoTmBtEempaenrdatHu,rSe0fo4 waeprperionxjiemcaitede,lyte2.samApfleebsottliensgwetrhee sMhTakBeEn phase,
ESERGOASZ
Faizotar
000044
DuPont EMSE Report No. 15.03
bthoetcrsimwpaesdtsraamnpslfeerbeodtttloes50wmrLe dpeoclaypprpoepdyalnendetcheenMtrTifBugEepchuabsees wriotmh eagclahsosfttrhaenssfaermppliepet T2ohf0temhMe.TMgBTlEaBspsEhsacpishneatislwelaartsioocmnenvctaarlcifhwouigftethdhaaetfcaiepnpitrnroiexfidumgacetatepulbayens2d0wt0ah0se RstirPnaiMnlslffaeotrr1ieod0nmwviiinat.lshwAgelra2se0smSpoiLpreaetltieaqtu1o0t. r20anCf.erFerdomweiatchha gfltahssepgilapsestsc0i.3laGtCiovnialviaalnsd, w2amsLseaallieqduowtitohftahleuMmiTnBuEm ipnhadseewrasp cap iafnolri1qG.5uCom/oLMfotShfeqmueaMntTthiaBfniEocalpthifoaonsroeMfwuao8.rs2nsaTetqBeuAde.nsteiFiadrlloaynmarleiyasecidsh.oinfathGe CglvaisslsucnidnteirllNastioanndvawla,s 6rmedli.ssolved
3.3.1.3 "sTTeuhmdep.teermpaetruarteurMeeofatshueretmeetnstysstem was monitored an recorded throughout th courseofthe
332 AnalticalMethodsforTes SubstanceandProducts 33.210 Analysis ofTest Substance
8-2 TBA Analysis:
Analytical standards: PTrhoedu1cHt,s,1HW,es2Ht,C2o.lpumebfilau,orSoCd)ecwaans-u1-soela(s3-t2hTBanAa,lytCiAcaSlHst6a7n8da-r3d9.7,Th97e.16H%,, 1OHa,lo2w,ood
2H-perfluoro-9-methyldecan-1-ol (C-11 Iso; CAS# 31200-98-3,98%, Oakwood Products,)
dh`weaapyslu1as4deeadcnaadlsudaaanryoi2nd8teecsmaaanmlopllste(aaMnnd+aalr)ydsi(sfD.or-d5Sat2yoTc0BksAsa,omlpuDtluiePosonsannt(d,10tw0ha0esm1ugDs,/eLd1)Da,o5f2ttDhh, i2anDnta,elmy3ta-il3casClt-ainndadradrfdor and the intemal standards were prepare in methanol and refgersted. Th calibration
standards were freshly prepared for each calibration in MTBE by dilution of the freshly made
2U550/.-L1m0og0f/0LCg-st1Lo1cok1s5f0in3f-om2retdThaBayAn.o0l.sCaomTnpylspeiascnatalnlvyd,eDlt-h3eo-fc2ailTnitBberAmaatlifoosnrtdsatanaydnadr1ad4rwdnsadswedaraeeyd:m2a8daspeapmrinpotlxhieem.raatFneoglrey d3oa0fy0 8905S2a(mT8-pB2lAeTa,BnAtdh)ienactnaedlmipbarelaatskistoarnncdcaurfrdo.vresAiwnoneremexizac.mo9pn5lse(ioCcf-t1ec1dal1u5i0sb)irnaagntidthotenhaecuirraovcieocwfoafttsheghipveceaonkncaiernnetFariafgtouriroeinoAsno2mfz c(fuoSrravqmeupsalnweteiTfrAiecSac0to-in3osn)trowufacsstaegdmipuvlseeinsngiTnhAFSeuOgr-ua1trietoooAfTS.AtheF5o0p-ed2a6ak.yaAr1a4chonrrodmfaodtnaoy1g/r22a3p3sh1aom(fp1ls2ea,mTpBtlhAee)sceiapbnardraaptteiiaoonkn airnatefmoarl sotnandmarzd.33A(nD-e8x.a2mTplBeAo)fancdaltihberaattioonocfutrhveecwoanscegnitvreantiionnsFiogfur8e-3A-T4BAfoarndthe oqfuasnatimfpilceatsieopnaorfastiaomnp(lSeasmTpAleS1T-A1S2t-o3T)Awa5s1g-i1v7aenndinTAFuSg2u-r1e tAo STA 52-17. A chromatograph
4Qu.a0n.t5ifmiLcatailoinquootf iss GC via (1.7
8-2 TBA:
omftLhevoMlTuBmEe)3p,h
a6se
LfroomftSh0e
GmCLvaolf(DS-e8c.t3ionT3A.1.i2n)erwmals
palnadcoerddiwnaas
Saadmdpeldetwhaseavniaallyuzsedint3wgicGeCbysyariGngCe/,MthS ivnisltrwuamsenctaapcpceodrdainndgstuobjtehctfeodlltoowainnalgysciosn.ditEiaoncsh:
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GOMS system: Column:
DHePte6c8t9or0 (PAlguisleGntC),(AMgPSi2l-eM)u,lHtPiP5u9r7p3osMeaSsasmpSleelrective (Gerstal) DB-SMS, 30m x 0.25 mm, | um film (Agilent)
Temp. ramp: Initial: Flow rate: Spit Inlet temp.
80C for 2 min 20C/minto 120C 50C/min to 300C and hold for 3 min 1.0 mL/min; He; constant low mode 51 250C
Injection volume: MSD transfer line temp. Tonization:
2uL 280C EL70eV
SIM ions monitored:
mz: 31,95, 131; dwell ime: 50 ms for each ion
(day 0 samples)
mlz: 31,33, 95,98; dwell time: 25 ms for each ion
(days 14 and 28 samples)
Retention time:
8:2 Telomer B Alcohol (C8-24):
497 min
Internal standard (D-8-2 TBA):
493-495 min
Internal Standard (C-11 Iso)
542min
Fluorinated acid Analysis: Analytical standards: The perfluorohexanoic acid (PFHA; CAS # 307-24-4, +98%, TCI America), hpeerxfaldueocraofolcutoarnoo-i2c-adceicden(oPiFcOaAc;idC(A2SHH-H3D3F5--26-7D1A,;97C%A,S#Oa1k6w1o0o94d-7Pr6o-d4u,cDtsu)P,on2tH)-, and 2perfluorooctyl ethanoic acid (2-PFOEA; CAS# 27854-31-5, DuPont) were used as standards for analysis of fluorinated acids in test samples. The analytical standard stock solutions for achofthe above acids were prepared at approx. 200 pg/mL concentration by dissolving appropriate amountoftheacidsin methanol. Asingle, mixed analytical stock solution at `approx. 200 ug/Lfoeach acid was prepared by dilutionofeachofthe above primary stock ssoolluuttiioonns(w~i2t0h0muegt/hLa)noalndnanasailnigqlueotvooflaumneetgraitcivfleascko.ntAronl aslaimqpuolteomfatthreixm(iTxreedaramceidntst5o)ckw.ere diluted together in asingle vial to produce approx. 30 pg concentrationof each acid in 10% diluted sample matrix in methanol. A separate 30 pg/L solution was prepared with a day 0 `nTehgeasteivtewocosnotlruotli(onSsamwpelreeTdeAsi5g0n-a1t5e)daansd30a duagy/L28stnaengdaatridvseacnodntaroplur(eSammeptlheanToAl w5a2-s15). designated as a0 pg/L standard. For fluorinated acid spike recovery experiment (quSaencitiifoinca3t.i2o.n6o),faincdailivbirdautailonusotrainndaatreddcuarcvide iwnatshecoancsitdr-uscptiekdedinsathmeplreasn.ge of 10- 50 pg/L for
EMSERIS-3@82
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Sample Analysis:
a`Alliiqquuoottssooffalllssaammpplleesswweerreetdrialnustfeedrr1e0dxtowiHtPhLmCetvhiaanloslfobrefdoirreectthaenaanlaylsyissisw.ithIonuatdddiiltuitoino,n. All s1p0exctdirloumteetdryan(dLCal/lMuSn/dMiSlu)t.edAssaemptloefsstwaenrdearadnsal(yszeeedabbyovlei)qwuiadscrhurnomaatttohgerbaepghiyn-ntianngdeamndmaatss cthuerevnedfoorfetahcehaancaildysciosnssterquucetnecde.usQiunagnsttiatnadtairodnswfarsomdotnhee buegsinanisiningngaglned ttwhoe-epnodiontfctahleibration analysis sequence. For maximal accuracyofresults, the results for undiluted samples for PFHA, PFOA, and 2-PFOEA were considered final. For 2H-HDF-2-DA, the results from 10x diluted samples were considered final. This was done to ensure that the peak area data valuesofanalytes in samples wereasclose as possible tothoseof 30 ug/L standards. For fluorinated acid spike recovery experiment (Section 3.2.6), a calibration standard curve was. `constructed in the range of 10 - 50 ug/L for quantificationof individual fluorinated acid in the acid-spiked samples. Chromatographsofsamples TAS2-3 and TAS2-6, , 30 pg/L of standard acid mix made in sample matrix TAS0-15, sample matrix TAS2-16, and a methanol solvent blank were given in Figure A-6, Each sample was analyzed twice according to the. following conditions:
HPLC instrument:
`Waters 2795 HPLC
MS instrument:
Micromass Quattro Micro
HPLCColumn:
Phenomenex" particle size
LunaC18(2), 20 mum
x
2.0mm,
3
pum
Mobile Phase: Mobile Phase Gradient:
A: 2 mM ammonium acetate in deionized water (18.2 MQ cm) C: methanol purge solvent; deionized water (18.2 M2 cm) needlewash solvent: isopropanol
Flow Rate `Time (min) %A %C (mUmin)
0
9 10
03
[5}
9%
10
03
15
0 100 03
45
0 100 03
46
9
10
04
65
%
10
03
Flow Direction Switching: 0t01.0 min ~ flow to waste
104.5 min- flow toMS
4506.5min -flowto waste:
Column Temperature:
35C
Injection Volume:
10uL
Tonization mode:
electrospray ionization in negative mode
Capillary volage:
350kV
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Ton source temperature: 140C
Desolvation temperature: 350C
Desolvation Gas Flow: Ny, 500 Uhr
Cone Gas Flow:
Na, 100 Uhr
Quadrupole Resolution: ~~ QI low and high mass resolution 10
Q2- low and high mass resolution 15.
Collision Gs, Pressure: argon, 3.5 x 10 mbar
Data acquisition mode: dmeulltaiyp0l.e0r5eascetcion monitoring (0-4.5 min), inter-channel
Dwell Time Cone Voltage Collision
Reaction (sec)
[4] Energy (eV)
PFHA
313>269 or
is
15
PFOA
413>369 ol
15
15
2HHDF-2.DA
457>393 ot
1s
15
2PFOEA
477>393 ol
EF
0
Fluoride Analysis Analytical standard Centified standard of 100 mg/L of fluoride in water (Thermo Orion ) was usedfor standard cdailliubtriaotnioonf. thTehe10c0almigb/raLtoiofnflsutoanrdiadredsstnaondramradllwyitwherTeISmAaBdIeIin t(thoetarlainogneicos5ftreng1t0h0audgj/usLtbmyent b=uff1e6r-I1;8)T. hHearlmfostOrreinognt)haTnIdSdAeBiIonIize(donweatpearrtforfomTIBSaAmBsIteIadpElu-sPuornee spyasrttoefm (deMieognoizhemd-cm water) was usedforthedilution. A 10 mi aliquotofeachofthe standard solution was used for fluoride analysis to constructa standard curve for quantificationoffluoride in samples.
`Quantificationof Fluoride: o`Tfhtehe1015mLmLtesptolmyepdrioupmyltehnaet twuabsestrweaastetdhwaiwtehdNaat OroHomantdeHmpieSrOautu(rSeecatnidonwa3.s3.c1e.n2t)riffruogmedc.ach Five milliliter from each ofthe tubes was transferred (0 a 50 mL polypropylene tube that contained 5 mLof TISABII solution. After mixing, the medium was analyzed for fluoride usian71g0 A Plus pH/ISE meter (Thermo Orion, serial # 066814) and a Ionplus fluoride. selective electrode (Thermo Orion, model 96-09, lot # GX1). After filling the reference. `icnthoamebaecrhowiftthheresfaermepnlcee meleedcituromdeanfidllfilnugosroilduetsitoann(daTrhdersmoloutOiroinona)n,dtthheeecloencdturcotdievwiatys iinnserted millivolts was recorded after minof incubation. Each sample was measured 1 - 2 times. A standard curve was generated by ploting the standard fluoride concentration in LOG scale: versus millivolts (Figure A-1) for quantification of fluoride.
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DuPont EMSE Report No. 15-03 40 RESULTS AND Discussion
41 Under the test conditions, 8-2 TBA was rapidly transformed (Figure 1 and Table A-1). Sp6i0k%eartedcaoyver1y4o,afn8d-214T4B=Af2r%omattdhaeyt2e8stm(TeadbileumA-a2)v,eriangdeicdat1i2n3g+tha%t MaTtdBaEy 0, 12=0 extraction method obtained a good recovery of 8.2 TBA from the test medium. Sp(iTakbelerBe-c1o,veBr-y2,oBf-f3l,uoarnidnatB-e4d).aciSdpsifoke rdeacyo0vesraymopfl2es-PwFasOEmAodearnadte2,H-raHnDgFe-d26.2D-A76% awbaosutpo3o6r%foarndday342%s,armepslpeesct(iSveelcyt;iosnpi3k.2e.r6,ecoolvle-rgyorofwPnFbOacAterainadl cPuFltHuArew,aasvemroadgeerdate, aivndeircaagteedd atbhaotutful1l %groawnnd 6ba%ct,erieaslpceuclttvureelyma(tTraibxlemaB-y1c,aBu-s2e,sBi-gn3i,fiacnadntBd). This wuenldle.restimationof 2-PFOEA and 2H-HDF-2-DA in day 14 and day 28 samples as + cAotmdpaayre1d4,wi5t7h0a%bioofti8c-2coTntBrAolhvaesssbeelesn(bTiroeraatnmsefnotr2m)e.d in test vessel(s Treatment I) + dAuteatyoa2c8o,m1b0i8n-a2tiTonBoAfwbiaostrdaetnescftoerdmaitnitoenstavnedssselbsi.otcThreemloovsasolfparent chemical was + lIonstshiesapboisostiibclcyocnatruosle,d bthyevcoolnacteilnitzraattiioonndofur8i-n2g tThBAincduebcarteiaosnedorcobnytiinnucooumspllye.teThe eixntorcaucltuimo.n due 10 adsorption to the lass wall ofthe est vessels and/or the sludge + Tfuhlely8a.c2ceTsBsiAbltehaftoradmseotrabbeodlitsomt,hweiwtahllmsorfetthheante9st0v%eosfseblisotdruarninsgfotrhmeacloiaotninatg dwaays 14 and 99%atday 28 (Table A-1).
4.2 hUnadtetrhteheadatpesttedcomnidictriooonrsg,asinginsimfsicwaenrtedeafbuloeritonamteitoanboolciczuer8r.e2d TduBrAing(0tlhueoersin,astuegdgaecsitdisng and other metabolites. + T64heugf/luLoratiddeacyon1c4e,natnradti7o0niung/tLesattvdeasyse2l8s,iwnhcirleeasteedfrreowmas~1n0o pingcr/eaaLtsedafoyr0t1he0biotic controls (Table A).
43 tFhoeurtesftlusoyrsitneatmedanadcihdemiertmabaoslsitbeasl(abnicoetrcaonnstfroirbmuattiioonns aptrdoadyuc2t8s)ahreavreoubgehelnyiedsetnitmiafiteeddfrom based in Table | and Table A-1: + The F(CF,),CF=CHCOOH (2H-HDF-2-DA)isthe most abundant metabolitesnd accounted fo at cat 10%of otal mass present. + ~F2(%CoFf)oCtHa,CmOaOsHpr(e2s-enPtF.OEAY i the second abundant metabolite, accounted for + F(CF)<COOH (PFOA) accounted for <2% oftotal mass present. + F(CF),COOH (PFHA) accounted for 0.4%of otal mass present. + TBhAelcfoohrmoalt[ioFn(oCfF.P)F.HCHA;CiHn,o0tHd,uCeAtSo #im6p4u7r-i4t2y.o7f)t,hwhtiecsthcchoeumlidcalle,adsitnocPeF6H-2ATelomer formation,wasnot detected in the test chemical (Figure A-7). The concentration of
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bPaFcHkgAroinucnrdealseevdelsoigfnPifFicHaAntlwyafsroprmedsaenyt 0in10thdeaaybi2o8tiicnctoensttrvoelsssedlusr,inwghitlhee otneslty. Defluorination of PFOA may lead to the formationof PFHA during the test. 4.4 Based on the metabolites identified, the following is a potential pathway for 8-2 TBA bbiioottrraannssffoorrmmaattiioonn p(arosdiumcptlsi)f:ied pathway which does not include all potential
RCEMCHON FR moos REY potion +E F(CF:;COOH +FF(CF:)syCOOH +4F-
A notable featureofthis pathway is that PFOA may be defluorinated (metabolized) to form PFHA.
50 CONCLUSIONS oUtnhdeerruntihedetnetsitfcioenddtirtaionnssf,or8m-a2tiToBnAproisdubcetisn.g AralptihdoluygthraPnFsfOoArmiesdotnoeoflfutohreinaitdeendtaicfiiedds and mmeettaabboolliitzeesd, oit afcocromunPtFedHAforan<d2%moayfnmoatsbsebaanlanuclet.imaPtFe O(sAtabalpep)armeenttalbyolmiatey ubnedefurrtthheertest conditions.
6.0 REFERENCE 6.1 3O0E2CC:D IGnuhiedreelnitneBifoodreTgerasdtaibnigloiftyC:heMmoidciaflise,d3M0I2TB:IZTaehsnt-(WIe)l(l1e98n1s).EMPA Test (1992);
EMSERIS-03/4842
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DuPont EMSE Report No. 15-03
FIGURE 1 8-2 TELOMER
B
ALCOHOL
(8-2
TBA)
CONCENTRATION
DURING
THE28-DAY
TEST"
cm
2
ii:LE
2
5
0
52
000051
_ DuDPuoPonnttEEMMSSEREeRpeoponrttNNoo..1155-0033
FIGURE 2
FLUORIDE CONCENTRATION DURING THE 28-DAY TEST*
--e-- --o--
AKidlalpedtebadctbearcitaelricaulltcuurletu+re2+m8M-2
TBA NaCN
+
8:2
TBA
& a
2g
S 2T 2
o 5 wo sw 3 ow Days after the Initiation
* dTehreitveesdtwfarosmitnhietioartiegdionanl19daJtuaolyf 2Ta0b0l2eaAn-d3finished on 16August 2002. The graph is
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------------ RIB DuPont EMSE Report No. 15-03
TABLE 1:
(E1S9T~IJMUALT-E2D00CA 2O)NACNEDNTDRAAYT2I8OS N(S16O-AFAUFGL-U2O0NR0I2YN)A.TED ACID METABOLITEeS ATeDAYe0
HoTsemearr
seroms _awiorzon pron ern
o Tosntts asgos oo hoy
wo ot wo
0
Tm
wr
"
TsoSimaeorrtstanaaecotnwonnaolh 00 ho
wo
wo
acs
oj0oey Tani comet on ovwmn ww wew oww aasws
+ "This table i derived from the orignal dstaofTable A, A-5, A-6 A, B1 B-2,B-3, and B4.
+ =SEs,t=imi%antSelpdicckooennccreeenncttorrvaaettriiyooonnf,otfCh,ein=diinCvdiiSdvu,iadwluhafellruelo:orriniantaetdeadciadcimde.tablies a day 0or 28 oDPfuFheOoA(r0iatanhdetelPdoFwHaAsip,diskteihnr%eteceotsvpseiarkmyepolrfeesc2.o-vPeFryOEvaAluaendwa2sHs-HeDdFfo-r2-cDorAecatnidnmgopdoesrsaitbelesuinkderresciovmeariyoonf
9 ND: Not detected above sample matrix background evel
BSERSOAsE
waren
000053
_ DuDPuoPonnttEEMMSSEREeRpeoporrttNNoo..1155:-0033 TABLE 2: `DTAHIELTYETMEPMEPREARTAUTRUERWEARSEARDEICNOGRSDIENDTHBEY LAACBAWLHIBERRAETETDHEDTIECSKTSOWNASRECCOONRDDUCETRED. (MODEL THDX, SERIAL# 0118247)
Time ony
Bl 2 3 . s s ' s 0 " 2 " 1 w 1s " a z 2 = = z Standard Deviation
[2 192002 202002 212002 2002 202 202002 2532002 2wa002 2r2002 22002 2202 02002 312002 hog 2002 2a02002 Shug2002 "av2002 Sa 2002 san Tavg.2002 sag 2002 Savm2002 10a2002 Aug2002 12852002 aga Teg2002 150200 16Au 2002
Temparaurs 20 2s as 21s 20 an 50 215 zs 20 250 0 as 2s 20 20
2m 202s 20 ns 20 20 zs 27s 2s us 202s us 0 os
EMSERI5-03/4842
~
Page 22047
000054
-_--
DuDPuoPnonttEMESMESREeRpeoponrttNNoo..1155:-0033
APPENDIX A:
EMSERI5-03/4842
Page 23 of47 000055
_--
DuDPuoPonnEtMESMESREepReoprotrtNNoo.. 11550-033
TABLE A-1.
ANALYTICAL RESULTS OF 8-2 TBA CONCENTRATION AT DAY0 (19-JuL-2002),
DAY 14 (2-AuG-2002), AND DAY 28 (16-AUG-2002)
TT No
Tmrovemant
We Time
coAnmvcseinytAriactailon
mAAvaeiryAlacgael
caoncamntateiont
T TAS SToosctamlenctutl-arAdnsgieeswe 0 1 w war omsr. aors
Tho IEmeTAdBiAunmpcoswe6d0se0w,no9fs1- 0 2T4 omoesm oe
Tout
ioa
os wm
=)
03
F--
Tasos
o me
E
2
os
om
os mo
"
076
Average &StandardDeviation (3 8)
som
seers
TASS mRTaardecseitrmpecnstr2-o2KdegrKoOuNtand. o 1] a5.1
no
2s
Ther BoonudseaBmIrTtBeAR 0 2Ima
Ed
1o6
eos Average & StandardDeviation 1 3)
TAZ Tnreascdmmnainnstdc-erAddasgGaerosiwtnh Tas013 bore Taso-14 Av&eStarndaardDegviseton (n=) ASIA SMTaardcshreammaoapntstur5e0A0dnyapotoeodfn Ts1z BiTAmcomsdmnn
bose sia asta srs Average &StandardDovition(n= )
1m 3 es
01 6 oz' whomt
2 ne a ns
3 nm 4 1: naeo 2 us
2 ma wos m2
a me wos aes
+ am wos ms
som
=
El
10m 158
wr
=
109
0
3
186
20250
8
27
no
10s
as
=
as
51
9
"
smassre
EMSERI5-03/4842
Page 24 of47
000056
_--
DuDPuPoomnEtMESMESREeRpeoporrtt NNoo. 1155:0033
TABLE A-1 (CONTINUED)
EE No
Trsmant Time coAnciaevsaton Arvaeygiecn cancantatont
r TS mma #1 r 9 sor = --
Tot SmoetoduumoranImKroNweS 1 2 aorm
0oug or62ToAR
;=
on
ar
sta condsaubt wom
=
ns
Average StandardDeviation (0-3)
sm
wozer
Tez nopmcrernccnaou-srpesdgsaersnn meta bo stra Avorsge&Standard Doviton (n=3)
61HE si woe ous
em woe aes
se
I
ost
a
a
I
1
reser
HA mez
fTmiSaaioemluannnodcnsg6sA30sioeipeedsnof
Soe
ees
mea
mes
[TCR------
1| o. Bm 2 0
: 5
se 24"0 ms oo
se
o
o
o
o
o
o
o
.
HS mez BO ws
Scrctrin ron
me
2
=
Ter moeudsmoprasI2MTKanCnNMW 2 :2 =a
w
-
Tike SUsgdem
Average StandardOmiaton (n+)
3 32m500
12
768.
srs
TsEMSEEmRI5Ee -03/e4842metreeergmrEprPra2z5g04er7
000057
-
DDuuPPoonmtEEMMSSEERReepporntNNoo.l1s5.00y3
TABLE A-1 (CONTINUED)
o TCona Tne cAroayma endritgtal concomraiont
TR mmementasipes B10wr w a ro --
ars msceasmmenncroemnGionn 3 310as a
maze butts
2s3 awo39m6
or
I.
Aeveeraeg--e S--andr--a e--ven------------------------as--a --
+ Standard Deviationwas calculated usin the Final concentration of TBA.
+ CFiunal8c.o2nTceBnAtraatnialoyntoifc8a-l 2avTerBaAg,e Cv;al=e,C, * (Vamee/V,) where: Va=30rmL (MTBE used 0 extract the test medium), Vi20mL (Test medium usd ro extraction).
EMSER15-03/4842
Page 26 of47
000058
-_
DuDPuoPonnttEEMMSSEERRseppoorttNNoo. 1155.-0033
TasLe A-2.
AMANTARLIYXTI(CTARLEARTEMSEUNLTTS3,OFADSAPPITKEEDREBACCOTVEERRIYALOFCU8L-2TUTRBEAPLFURSOGMRTOHWETSHAMMPEDLIEUM IN
COATED GLASS SERUM BOTTLES) AT DAY 0 (19-JuL-2002), DAY 14
(2-AUG-2002), AND DAY 28 (16-AUG-2002)
TT no
Tee Re CoAmnricvaRincnslon veAneairRyceal E ConcanT iatont wRoceortsyt
T wr m wer er
mes 0 1 eewn PA a
"
mow oz a os : er
1
meno 3 em 2 ee
as
ns
Average: StandardDeviation
mas
mes ow 1 am Te
"
ao
n
meow 2 somw = 2)
ww 2 2
wwmm
-
a
=
Average Standard Doviation
nose
mes aw 1 wmee P os
we
maw wz om PE
0
mo
ws
man aw 3s om wr
aso
Average
sm StandardDeviation
az
+ Final 8-2 TBA concentration, C; = C, (Vyrae/V.) where:
C= 8-2 TBA analytical average
Var = 30 mL (MTBE used to extrac the test medium) Vo= 20 mL (Test medium used ro extraction) 1%ofspike recovery, R = (C-CVC.] x 100, where: Ce= Final 8.2 TBA concentration in th spiked bottles C= Final 82 TBA concentrationin th coated bottles without adding or spiking with
8-2 TBA stock solution (Treatment 4)
C= Concentrationof 8-2 TBA spiked into the sample bottles (600 gL; Treatment 3)
EMSERGOA8Z
Pa2g7 eof47
000059
DuPont EMSE Report No. 15-03
TABLE A-3.
DANAAYL1Y4TI(C2A-LAURGE-S2U0L0T2S),OFANFDLUDOARYID2E8 C(O1N6C-EANUTGR-A2T00I2O)N
TTT o
Trosmamnet
Te We coAnFmcaeuincveaatlon
wm
wr
TSO STeawdcsemtnenctes6-n0a0gg3ge1soof
0
1 1
sa se
Tea ITeAncvedsn 0 2 4s
EY
PE
eos
0 3 as
3a
Tasos
0 4 as
AT DAY 0
wFApaoaivgoicelal wr se a a a
(19-JuL-2002),
Fconcuamiraetont wr 08 0 a os
0s Average + SandardDeviston(n=
TASS SrTreasettmeanotu2n-ZirtgdRsand mer EComneudasasnmaoTteem hens
Average Standard vation n=) HL TSnusunmimeln5ctstra50d0gayreowdotofh5 sta IoTeoAnewessan mrs
mre
0 sar ss
o 1) ra 0 2: 1s ose
3
1| wz: ms wos ome
3 w oa me
0
00
aston
"
ws
78
ws
76
2
11208
ut
we
2s
0
En
@z
EY
se
is
Average StandardDeviation (n+ TASKS TnSeraeemnsioc2muiauKnordtgArowad sir GCooevdgsoosmaoToram
AvsteeStrandaardDegviateion (n+)
wos ome s
wo. re Ww :2 es wos es
2
as
seten
6
ns
6
6
arsBees
EMSERI5-05/4842
--
Page 28 of47
000060
DuPont EMSE Report No. 15-03
TABLE A-3 (CONTINUED)
-- No.
TTryopmemoanlt
TASZY bTarceieartamlecntur1-nAdrgtoedw Tazz 2meTdoiAummcpouswe60d0sJenLmof 8-
poe 23 Ths24 Ths25 Average +StandardDeviston (n=5)
Rap Time. wm 28 1y
28 2 2
EE 3
EY
El 5
TASZS mbTaroceidaeutnsmpecunu2str-2eKiMilneraKOoNwand. EI . TAT cSoOatgedLsoonIu2m TboBttAe M 2 2.
Ths28
23
3
Average + StandardDeviation (n =3)
AFmliugotricdael Concentration"
wr h6i0d u3 us u3 0 1 74 as 20
os se 50
FAniauioyriicdael Avarage mr 88 13 uz 6s ar
6s 54 so
coFnicneanltFrlauteiniodne; mr ne os 4 ns 74
nazs0 no 08 100
nase
+ Standard Deviation was calculated using the final fluoride concentration. + Final fluoride concentration, Cr = Cy x [(ViHVO/V,] where:
C.= Fluoride analytical average value, V,= 5 mL (TISABII buffer added to the test medium for fluoride measurement), Ve=$ mL (Test medium used for fluoride measurement).
EMSERI5-03/4842
Page 29of 47 000061
_ DuDPuoPnotntEMESMESREeRpeoponrtt NNoo. 11559-0033
TABLE A4.
AETNHAALNYOTIICCAALCIRDES(U2L-TPSFOOEFAF)LUAOTRDIANYAT0ED(1A9C-IJDULM-E2T0A0B2O)L,IATNED2-DPAEYR2F8LUOROOCTYL
(16-Auc-2002)
TT No
Trmosemant Time ew coAnIncseinmytriamctailon iAAnvaweirgotaigBceal cFonacanmtrestornt
TAZ
iTartmsentrsowhAdmspaisaubapcuessal
wm 0
1
wr no
wr no
mr -- rot
TAD 0Sa0ru4m0Sooree 2TEA costed 2: wwoo no
Average (n42)
2 "0 wo
TASS cMTrtaKtrmCei ntra2on-dwK6lm0o0s0eb1acag1dru6ss.l2 01: no
TAB TBARmcowadsenmece 0 2 no
Avwrage (102)
oz o
no
o
no
no
"o
TASZ2 u S0VTroinsoofe15tu2ptTdmBamApateneidcmoncaleoattsal 28 11 p4y 15s se
TAS23 sen boti
2 21 24 ae
Aveage (0)
2 zs 2
TASS cTrieKsamCeNntagr2no-dwKt6ihlme0abms0c1osp18aols2 28 15 " no
wo
TAsz7 TeAmcossdsnmbote 2 2 o
wo
no
AAwvworaggra(ener2)d
2 no owwo
++ NFiDn:alN2o-tPdFetOeEcAtecdoancbeonvterastaimopn,leCymat=riCxyba[c(kVgargoruend/Vle/v(elV.u/V)] where:
CVaur=ry2e-=P3F0OEmALa(naMlTytBicEalusaevdetroaegxetvraalcute,the test medium),
VV,==260mmLL((TTeBstEmephdaisuemdruiseedd ffoorr meexttarabcotliiotne),analysis),
Va = 1.5 mL (Methanol used to redissolve the dried MTBE phase for metabolite analysis).
EMSERI5-03/4842
Page 30 of47
000062
_--
DuDPuPoonnttEEMMSSEEReReppoorttNNoo..i155-0033
TABLE A-6.
AAcNiADLY(TPIFCOAAL)RAETSUDLATYS0OF(1F9L-UJOURL-I2N0A0T2E)D,AACNIDD MDEATYA2B8OL(I1T6E-APuEGR-F2L0U0O2R)OOCTANOIC
I No Trost Time coAnncaantiavnion_nAvrryeiegael concamant
TASO2 Trre ruo1utAtsmampooaddtuancoartsa6l00 wm ;1
wNro o>
anro
wr ot
TasoSSSoLera S TEA ncseom zw ro wo
Average (102)
: ow wo
TASSTmaeimeenntgr2oKubtoacodssols 2m TAO CKoCNeadnsdo60m0pboleof 82 TBAin Avrsge (02)
0 1 wo z 1 w" oz ow
"o
wo
wo
~o
TAS22 iLTBmLroerneE2T1tiEsApdmnamgcraoadstanocsoaotsria$lr00 28 1Tr oaors as 03
The2a Boe
woz om ss os
Average (1-2)
2: me "00
HLS Tmt ses 31 ho
TAS27 cKooumdsaerumbiee an a 2Towow
Average 1-2)
: we
"o
no
wo
"o
wo
+ND: Notdetesbcovetsaempdlematrixbackground level + Final PFOA concentration, Cy = C, * (Vase VV, Va) where:
Cy= PFOA analytical average value, Var= 3m0meL (TBE used(0 extract the test medium), Vo= 20mL (Test medium usedfor extraction), VV,=a=61m.5Lm(LT(BMeEthpahnaosleudsreidedto froerdmiestsaoblovleitdtherainaeldysiMs)T,BE phase for metabolite analysis).
EMSERISONARZ
Pageirord 000065
DuPont EMSE Report No. 15-03
TABLE A-8. (COMPONENTS OF BACTERIAL GROWTH MEDIUM USED FOR THE TEST.
KHyPOy NaHPO*H20 Kal MgS047H20
CaCLy*2H0 NaCl FeS04*TH0
CuS045H0 CoClp*6Hy0 MaCl*4Hp0 NagMo0#220 CaHsNaz07*2Hz0 (Sodium citrate dihydrate tribasic) NiClp*6H 0
ZnCl `Yeast Extract
Final Concentration 07081 03951 0581 0sg1 002591 10g 05mg 005 mg 01mg 0.01 mL. 0.025 mg Img
0.1 mg 0.005 mL 201
PH=72
EMSERIS-O3Asaz
Pageddorar
000064
_ DuDPuoPomnMtEEMMSSEERRseppoorttNNoo.I1s5-0033 FIGURE A-1 A FLUORIDE STANDARD CALIBRATION CURVE
2 20 = F2 s . a3 u 2. 23S os os wo
F~Standard Curve (for TAS1, 8/2/02)
=46403 001811 (=0.998)
IY =Log(ppXb=m)v;]
wo
To = 0
Millivolts
EMSERSOASZ
Fawera
000065
- OuonEMSERgmDuPNont Eo MSEl ReportsNo. e 15-03 FiGUREA-2 ATAA'C5A0L-I1BR7AT0ITONAC5U0R-V2E6 USED FOR 8-2 TBA QUANTIFICATION OF SAMPLES
[J -- --1
|
|
SAR TBA: 25.1 4020 pg. Calibration curve used for samples TA 50 - 1:26. The concentrationofthe
intemal standard: 300 ug/Lof C11-iso; the rangeofconcentrationsof8-2
[EMSER15-03/4842
Page 36 of47
000066
m CL mseemoe ut ESs E Repos1508
FIGURE A-3 A CHROMATOGRAPH OF SAMPLE TA50-3 USED FOR 8-2 TBA ANALYSIS
po
---a- ||
mass
=-- i"
-
1200 ||
a=80 |
|iA
- Ty
Tei TH TTRE a
`Sample TAS0-3. Quantification was done using peak areaofion mz 95; retention
time 4.97 min = 8-2 TBA; retention time 5.42 min = Cl1-iso.
TE
000067
_--
DuDPuoPonnttEEMMSSEEReReppoorrtNNoo..i1s5-0033
FIGURE A4 A CALIBRATION CURVE USED FOR 8-2 TBA QUANTIFICATION OF SAMPLES
TA 51-1 70 TA51-17 AND TA 52-1 To TA52-17
yd
||red
i
| re
|
3
1
i
i
{od
~
|
]
|
:
i
nT
J
Calibration curve usedforsamples TA 51 -- 1:17 and TA 52-1:17. The
ccoonncceennttrraattiioonnosfotfh8e-2inTtBerAn:al2s5t.a1nd--a1r0d0:551pg4/Lu.g/Lof D-8-2 TBA; the range of
EMSER15-03/4842
Page 38 of 47
000065
_ DuDPuPoonnttEEMMSSEERReeppoorrttNNoo..1155--0033 FIGURE A-5 AA CHROMATOGRAPH OF SAMPLE TA 52-3 USED FOR 8-2 TBA ANALYSIS
Abundance
12000 | 10000 8000. 6000 | 00 | 2000
on31.00(30.701031.70): 091602690
hy ime-->
4r 0 45 r 50 55 60 65 70 75 80 85 90 95 100
Abundance
"9 on33.00 (3323707)090 16020590
12000 10000 ao00 | 6000. 4000 2000
0% 40 4+5 50 55 60 65 70 75 80 85 90 95 100 ime-->
Sample RT 4.93
TAS2-3; lon 31, retention time 4.95 min min = D-8-2 TBA (internal standard).
=
8-2
TBA;
lon
33,
EMSER15-03/4842
Page 39 of47 000069
_ DuDuPPoonmt EEMMSSEERRepoortt NNoo. I1s5-03
OFFIGSUTRAENAD-A6RDCAHCRIODMMAIXTOMGARDAEPIHNSSAOMFPSLAEMMPALTERSITXAT5A2S-03-1A5N,DSTAAMSP2L-E6,MA3T0RIiXc L" TA52-16, AND A METHANOL SOLVENT BLANK. TAtr$e2.s6 wArbeintscontra, Trem 2) oro, w--T--A $--23--(Test sample, TreaJtmenta 1) --
mere
ee
he . I ven RE Wa------n
py
tl
STE
me
laeranemnla] gp]FAIA ne
Jr mT TEE wa 1 aHm mm ner]Eaan
1 TE
|
FE&He]EE_
Standard:TASD1S Sample mats, Teamens)
+30 pugL* acids.
.
=
-
=
TA S116 (Sample mates,Teme
J
7
PIE re aon a
-
=
1.
mE
CsnRFWR PenCamaemA
.
Foy |
1EVw ian7y3p nn
BERGA
Fader 000070
- ormoseeneESEs Beprn . 153
FIGURE A-6 CHROMATOGRAPHS OF SAMPLES TAS2-3 AND TA52-6, 30 poL
OF STANDARD ACID MIX MADE IN SAMPLE MATRIX TA50-15, SAMPLE MATRIX TA52-16, AND A METHANOL SOLVENT BLAN--K CONTINUED FROM PAGE 42,
Het vk
A go wd ws
] wn renner
ER
fe
000071
_--
DuDPuoPonnttEEMMSSEERRpeopomrNt oNo..I1S50-033
F8-I2GUTRBEAA-T7ESAT SGUCB/STMASNCCEHRUOSMEADTIONGTRHIASPSHTOUFDYMATERIAL CHARACTERIZATION OF
10c0re. hd
a
3
sso
wa 8278 0020)
se
CFL(CR=CHCH,O0H (03%)
.
+
i
ESERGOAS2
Pega 000072
_--
DuDuPPoonntEEMMSSEERRegpeormt NNoo. I1S5-0D3
APPENDIX B:
EMERSAsON
Faederd
wou 73
-_
DuDPuPooMntEMEMSSEERgReppoorttNNoo.1s15-303
TANAABLLYETBI-C1A.L RESULTS OF SPIKE RECOVERY OF 2-PERFLUOROOCTYL ETHANOIC
ACID E (2-NPoFOEAT)imAeTDAYRem0 (1c7oA-ncSeeEmPvne-t2on002a ),hwAeeNrnDe DAYo2n(e1n9a-nSEtP-20e02n)o.n
E mes 5 .a 0i57 C To - n
mes 0 22 wnro ws
a
"
mes osa wwrn on
2
`Av & Ste andr ard a Devigatie on?(n=3)
wre za Py ws
7622
ws
wis 2 2: wwos as
ws
n
mee zs> saarss sas wa
a
Ae vro StandDeviation (n=e ) eres r sess eer
+ Standard Deviation was calculated using the fina2-PFOEAconcentration.
1 FinCal=2-2P.FPOFEOAEcAoanncaleynttircaaltaivoCenr;,ag=e vCa,luxe,(Varse /V,) where:
Var= 30mL(MTBE used t0 extrac he test medium),
V.= 20 mL (Test medium used for extraction).
% ofspikerecovery,R= (CC) * 100, where:
C= Final 2-PROEA concentration in the spiked bottles,
= Concentorfa2-tPiFOoEAn spiked into the sample battles (200 ug/L).
EMSERI5-03/4842
Page ddof 47
000074
-_
DDuuPPoonNt EEMMSSEERRegpoortNNoo.1s150-033
TADENACABELLNYEOTBII-CC2A.ALCIRDES( U2LHT-SHODFF-S2PI-KDEAR)EACTODVAEYRY0O(F172-HS-EHPE-X2A00D2E)C,AFALNUDODRAOY-22-
(19-SEP-2002)
RC
A SriEmi2ia
concent
ST
pocorn
T meso a wnra w wAr mpra o
wes 0 2'
ws wo
as
p
"
wes os2
poss ass
ss
w
n
5 50
Average StandardDeviation? (n=3)
813
wes za. saan ws se
wis 2 2: "w0ss Py ons
n
mes 2s5 spass so
ar
"
Aev Standd ards Dois n (+3) --------eeerertuese es
1 Standard Deviation was calculatedusing thefina 2HHDF-2-DA concentration.
+ Final 2H-HDF-2-DA concentration, Cy = C, x(Vag/V,) where:
C= 2H-HDF-2-DA analytical sversge value,
Vira=30 mL (MTBEusedt0 extratcht test medium),
y= 20 mL (Test medium used or extraction).
C%=ofFisnpalik2eHre-cHoDvFe-ry2,-DRA=co(Cnc/eCn,t)rati1o0n0,ntwhheeres:piked botes,
C, = Concentrationof 2H-HDF-2-DA spiked into the sample bottles (200 ug/L).
EMSERS OAR
madera
VOLVO'S
DuPont EMSEReport No. 15-03
TABLE B-3. ANALYTICAL
RESULTS
OF
SPIKE
RECOVERY
OF
PERFLUOROOCTANOIC
ACID
(PFDOAA)AAT DA0Y (17-A SEP-200s2p)r,ioAxeAsND DAYAFr2AoaR(i19e-SeEcPFa-2cs o0a0Ro2) s Kars
e No Tmomer nw cownsmremten e mvnrwo wT econ}
wea 0 1' (wAr ws
nn
o
wes oz2 w0es B
ws
o
mes oo: was
us
n
AvansandardDoin 103)
wiz
wea 2 ' spoe 0
ws 2 2: ss0s ws
wo
wee 2a5 py PY
ns
Avwage StandOoatn (1-3)
nen
+ -- Sandar-- A d Deviae tion wase clculr ted usi-- n thefie nal2-Pe FOA cone centratie on. es
4 Final PFOA concentration, C = C, (Vame/Vo) where:
C= PFOAanalyticalaveragevale,
Vers=e 30
Vo 20mL
mL (MTBE used to
(Test mediumused
oexrtreaxcttrtahcetitoens).t
medi
um),
% ofspike recoRv=e(rCyC,, x 100, where:
C=Final PFOA concentration inthespiked bottles,
= ConcentrationofPFOA spiked ito the sample bots 200 g/L).
--EMS--ERI--S0g3--E/48m82 ---------------S------------Pea46rgofe4z7
voLL76
_
DuDuPPoonNtEEMMSSEERRpepoorntNNoo.I1s50-033
TABLE B4.
A(NPAFLHYAT)IACATLDRAEYS0UL(T1S7-OSFEPS-P2I0K0E2R)E,CAONVDERDYAYOF2 P(E1R9F-SLU?O-R2O0H0E2X)ANOIC ACID
TT e w No Tm ew r cmwcreen m aeme m e e oad
wes 0 . T7oss ma
i
wes oa2 waor oe
w
Bl
wes oaa anrr 2
o
Average StandardDeviation (n=3)
6223
wre 2a' 0nss ne
wo
wis 2a2 waer ws
ar
"
mes zs5 a0ne wor
"
_Averoge SansDntin (n=)
war
Standard Deviwaasctaliculoatned using the inal 2-PFHA concentration
1 FiCna=l PPFHHAA acnoanlcyteinctalraatvieorCan;g,e=valCu,e,x(Vaqrae Vi) where:
Va=r30mnL (MTB used 0 extractthe test medium,
Vi 20mL. (Test medium used for extraction).
% ofspike esovery, R = (CG) * 100, wheres
= Final PHA concentration nth spiked bot,
= ConcentrationofPEHA spiked ito the sample bes (200 pg),
EMSERISOVASZ
hamden 000077
