Document wrKekee68RRMDK45ggnp0N4gJ

: AR 236 - 1263 _-- DuDPuoPonnttEEMMSSEERReeppoornttNNoo..1133:-0033 Study Title READY BIODEGRADATION OF 8-2 TELOMER B ALCOHOL (MODIFIED OECD 301D CLoseD BOTTLE TesT) Test Guideline COrhgeamniiczaaltsi;onSefcotrioEnco3:noRmeiacdyanBdioCdoeogpreardaatbiilvtey:Dev30e1loDpmCelnotse(dOEBCotDt)eTGeuisdtel(1i9n9e2)f.or Testing of Author Ning Wang, Ph.D. Study Completion Date 20-March-2003 Test Facilities (ECLentdraulPRoensteadrecNhe&moDuervselaondpmCeonmtpany. `ECnovriproornatmeenCteanltearndfoMriEcnrgoibnieoelroignicgaRleSsceiaerncches & Engineering NGelwaasrgko,w DBuEil1d9i7n1g4-36001,01P,.OU.SBAox 6101 and HE.LaskdelulPLoanbtodreatNoerymofuorrsHeaanldthCaonmdpaEnnvyironmental Sciences Newark, DE. 19714 Submitter `E.D1u.PdonutPCohnetmdiecaNlemSooluurtsioannsdECntoemrpprainsey Wilmington, DE 19898, USA EMSE Study Project Number 1303/4842 Report Number EMSER 1303 EMSER 3-03/4842 Co Pagelor Co. AlN No csi 000005 DuPont EMSE Report No. 13-03 PAGE RESERVED FOR SPECIFIC COUNTRY REQUIREMENTS EMSER13-03/4842 Page 20128 000006 DuPont EMSE Report No. 13-03 (CERTIFICATION OF AUTHENTICITY READY BIODEGRADATION OF 8-2 TELOMER B ALCOHOL (MODIFIED OECD 301D CLOSED BOTTLE TEST) We, the undersigned, declare that the work described in this report was performedunderour supervision, and that this report provides an accurate record of the procedures and results. Report by: 20 cnr SeniorNoRgewsaegarBchDBiolog bu 03/30/03 Approved by: ] ye AET. Gannon, Ph.D. 03/30/03 Date 2S6tuDdey cIneitmiabt2ie0o0nr2Date: 20D-aMtaerScthud2y00C3ompleted: SB1uu.bbmdoiutntPCeohrne:tmdiecaNl SeoumonaosndCEunormrppraisnsey Wilmingion, DE 19898, USA EMSER3-03/4842 Page 3of 28 000007 DuPont ESE Report No. 13-03 TABLE OF CONTENTS CPaEgMeCAReUsieOrNveOdFfoArULSNpeEcRiIfCicILCYour ntry Rv EQUIs TEMENS...........cunmnonssssnsnricndd Tableof Contents........... C--O spoon) 2.0 3.0 GMenaertaleSaHrnAdiYMIEatNlhOOsGMS.McBccIceOcNec.e.ve.rscrorsrrssnrsnssenssrsnmssmssssssnssnsnnsnnnsssnsssoss] 3.333S.a1mplSeamEpXlTeACCHoOllNec1tionAnRdBIEYXISTIASC.O.N.v ..rve rrsrr rsnns ssnsssnnnn0 s]0 332 Analytical Methodsfor Test Substance and Products ...........rerecl| 4.0 RESUSaN DISCUSSION... |2 30 Condisliummmmmmmmmmmmmmmm----------_3 6.0 TB. LBimAitRatiUons OG f the 1m et.s c.ssssnsrsvsvsnssosms-- nsn-- nnno-- non-- ssis3 c 13 Figure | 8:2 Telomer B Alcohol (8-2 TBA) concentration during the 28-day (est*......|4 Figur2e. Fluoride concentrationduring the 28-day teSt.....ovvvromnininrarinnnn 1S Table(1| 4-CJoan0c-e20n0t3r)a,tiaonndodfa8y-228TB(2A8-atJada1y-200(0331)-Dec-20c 02), day 7 (v 7-Jan-2003), s day 14 16 Table2 Daily temperature readings with a calibrated digital thermometerinside a chemical hood where the test VesselsWere INCUBBIEd ......rorovvvrorornnnec|1 TableJAa-n1-2.0A0n3a)l,ydtiacyal14re(s1u4l-tsJaonf-82-0203T),BaAndcodnacyen2t8ra(t2i8o-nJaatnd-2a0y003)(.31.-cDecc-r2c00r2c),rdcacyn7r(r7-sn 19 Table(AT-r2e.atAmneanltyt3i,caacltrievsautletds solfuSdpgiekfeilrtreactoevpelruys omfin8e-r2aTl BmAedfiruomm)tahtedsaaymp0l(e3m1a-tDreicx- 2002), day 7 (7-Jan-2003), day 14 (14-Jan-2003), and day 28 (28-Jan-2003) .........21 Table A-3. Analytical resultsoffluoride concentration at day 0 (31-Dec-2002), day 7 (7- Jan-2003), day14 (14-Jan-200a3n)d, day 28 (28-Ja1-2003) c.crrcrrsnin 2d Table A-4. Preparationofthe mineral Medium fOr th (e5t.....vvrvrororvenrrnnnin25 Figure A-1 A fluoride standard calibration curve used for fluoride quantification of SAMPIES EIIIBAEI-T10 EIIIBEI64 crs 26 Figure A-2 A calibration curve usedfor 8-2 TBA quantification of samples E93384EJ-1 Figure0AB-3OSATcEhRrDomEa.t.og.r.a.phcoofossammpolnesEs9e3n3e84sEcJo-2s9msusmeednfmorm8m-m2 eTmBmAmmanmalmymsimse.m.s.s.s.s.s.s.s.d28l EMSER13-03/4842 Page 4028 000008 DuPont EMSE Report No. 13-03 READY BIODEGRADATION OF 8-2 TELOMER B ALCOHOL (MODIFIED `OECD 301D CLoseD BOTTLE TEST) Author Ning Wang, Ph.D. 10 SummaARY Rationaleofthe Study "eTnhviisrtoentmegnetnaelrapteerssiesntveinrcoeonmfeanttealtfsatuebsitnafnocrem.atWihoernelaevcahnetmtiocaaslseentsertshethpeoteennvtiiarlonfmoernt, cbhieomdiecgarla.datTihoen"isOoEnCeoDft3h01e DmaCjloorserodutBeostthtatTdeestt"eirsmianewsidtehleyeancvcierpotnemdentteasltfoataesosfestshtehe "thriesatdeys"tabgiiodveegrnadcahbeimliictayloisauftnelsitkeclhyemoicable. peDrsuiestteontthien stthreienngevnitrotnsmtencto.ndiBtieocnasu,sie atctpiavsasteesd smlauydgoe cwilclifbuethruissecdheamsitchaeliennotcerusluamdfoomretshteictesse,wtahgeeoturtecaotmmeeontftphlantte(sPtOwTilWl)i.ndiTchaetelwohwawtater s`oTlBuAb,ilCitAy,Svo#la6t7i8li-t3y9,-a7n)dtosbteronugsesudrifnactehiasdtseosrtprteiqonuiprreoapsermtoideisfiocfa8t-i2onToefltoheme3B0r1ADltceoshtol (8-2 e`gxupiedceltiendest.o bBeecualutismeattehelydumreattaiboonloifztedh,ettiesstsipsecriefliactisvteluydyshwolr (o2n8lydapyrso)viadnedi8n-f2orTmBatAioins onnot primary biodegradation potentialof8-2 TBA, through measurement of parent loss and bfioordmeagtriaodnoatfioflnupoortiednet,ianlotofthfeluuolrtiinmaatteedecnhveimriocnamlesnstuaclhfaase8o-f2tTheBAte,stachteesmtiscayls,temTowiatshsess full madeadpituedmmpilcursoaodrdgiatinoinsamlcsatrobtohnestoeustrccehe)mmicaaylbaendneaedfeavdorable growth medium (mineral Test System: "ATlhceohbiooldeingrmaidnaebriallitmyeadnidubmioptlruasnascftoirvmaatteidonslpuodtgenetiianloocfultuhme (t5estmsLubascttainvactee8d-s2lTuedgleompeerr lBiter of cmriniemrpaeldmteesdtivuemss)elfsro(mglaasPs OseTrWumwbaosttdleetse)ramnidnetd.h tTeshtewtaesstcsoynsdtuecmtecdonastisrtoeodomfteimnpdeirvaitduuarlely (W=i2t2hCa)c.tivTatheedssaltuudrgaeteadnsdolkuetpitoniocfl8o-s2edTeboltotmleesri8n tAhlecdoahrokl aitn rmoinoemratlemmpeerdaituurme.waPseriinoodciuclaaltleyd (exdtaryacst,io7n,a1n4d, aannadly2s8i)s,. saPomtpelnetibaolttdleegsorafddaitfifoenreofnttheexptesrtimcehnetmailcatlreaantdmefnotrsmwaetiroensoafcproitfeincetdiaflor `temsettacbhoelmitiecsal(tarnadnsbfyoramnaatliyosnisprfoodrucfltuso)riwdaes(fFoollno)weddubryinagntahleys2i8so-dfstthuedyc.oncentrationofthe Findings: a`Unndde1r3t%hemtoerstecaotnddiatyio1n4s,ctohmpa10rsesodfwi8t-h2tTheBaAbifotriocmctohnetrtoelstvveesssseellss. wAatsd~ay8%28,motrhee at day 7 ccoonntcreonlst.ratNioonsdoefflu8o-r2inTaBtiAonwoefre8-n2oTt BdiAstwinagsuiosbhsaebrlveebdedtuwreienngtthhetteets.system and the abiotic EMSER13-03/4842 Page 5 of28 000009 ---------- SEMIN-- p Ne,10 Conclusion: Underthe test conditions, 8-2 TBA is not readily biodegradable. + Confirmed expectation that abioic removal mechanism--s volatility andlor awdistohropbtjieocnti~vewoilflabcehifeacvtionrgs tmhaastsnebealdacnocnesideration when designing future studies 2.0 GENERAL STUDY INFORMATION Study Objectives Dectoencremnitrnaetitohnedbuiroitnrgantshfeoersmtation potential of8-2 TBA by monitoring its Determine the degreeofdefluorination of 8-2 TBA Deftaectromrsitnheaitfmaabyioatfifcecrteamboivlaitlymteocahcahniiesvemsma~vsoslabtaillainctey.and/or adsorption - will be: TestSystem Justification "The test system is outlined by the OECD 301D guideline and readily accepted. Study Personnel E.L. +duPoCennttdaelNReemsoeaurrcshaannddCDoemvpealnoypment- Corporate Center forEngineering Research - HaEsnkveilrolnmLeanbtoarlataonrdy MfiocrrHoebailotlhogaincdalEnSvciireonnmcee&nstaElnSgciineenecreisng Management: Study Director: JElo.hndTu.PoGnatnndoen,NePmho.Du.rs and Company CCeonrtproarlatReesCeeanrtcehr afonrdEDnegvienleoerpimnegntResearch EGnlvaisrgoonwmeBnutialldianngd3M0i0c.rPo.bOi.olBoogixca6l10S1ciences & Engineering Newark, DE 19714-6101 USA aS.ndMark Kennedy, Ph.D. EH.aI.skdelulPoLnatbdoreatNoermyofuorrsHeaanldthCaonmdpaEnnvyironmental Sciences Newark, DE 19714 USA NE.iLngduWaPnongt,dPeh.NDe.mours and Company CCoernptoarlatReesCeeanrtcehr afnord EDnegvienleeorpimnegntResearch GElnvaisrgoonwmeBnutialldianngd3M0i0c.rPo.bOi.olBogoixca6l10S1ciences & Engineering Newark, DE 19714-6101 USA EMSER13-03/4842 Page 6028 000010 DuPont EMSE Report No. 13-03 Avil Chemis: Techical Persone FBio1sgkdedalunlPoaSnuhtosdoiehr,NePamAfoDou.rsHeantdhCaonmdpaEnmyromenal Sciences Newik, OF 19714, USA aKetitih.Wc.rcFkoelns,om,DuDpuoPnotntHaCsekneaLRabeorsa&eDeaverlopcmehnt SteuxdpyerExiemceuntiSonrDaDtaets: 31-Decamber:2002 EStxupdeyriCmoemnpalletCiomopnlDeattieo:nDt: 2208-JMaarcyh22000033 3.0 MATERIALS AND METHODS 1 Tetsysem 31 TNSesyatnmSoeun:bysmtance (0F2eTreutoormoeroc8yAeltcaonhoel, 2 T5A CAS Name: Active substance(s) $N8D-3e2e4cT8pen5lee5om6,ae6rn1B1:8A8lc9o9ho.l1,09.91%0.10- SCMoAtlSeecNutul:amrbweerisgh:t oman 464.12 gmole" LR . F OH FAX CF FC NNT . F.U. N C. e TF LEoMtSNEumSbaemrple Numb: rE9o3v3e8r60 CCoonncceenoofnfaaa3sst,.naiiolmyiooznaenlnd:: 9999.%3% Mor impurity 08Ci%fC0F-C8HCH8OM EMSER13-03/4842 Page 7of28 000011 DuPont EMSE Report No. 13-03 Certificate ofAnalysis Date: Date Received: Solubility at 25C. Vapor pressure: Stability: Appearance/Color: Storage Conditions: Safety Precautions: 13-5ept-2001 26-Mar-2002 ~140 gL" 0.023 mm Hg Stable at ambient room temperature White solid Room temperature; keep tightly closed Wear lab coat, protective gloves, and safety glasses 3.02 Reference Substance None 313 PreparationofMineral Medium (OSneeedTaayblbeefAo-r4e ftohre miininteiraatliomnoefdithuemtsestto,ck2 msoLluteiaocnhporfepmairnaetriaoln)stwoecrkesaodludteidontsoA2,liBt,erCs,oafnd D sftielreirleundietis.onized water and the mineral medium was sterile filtered into two Nalgene 1 iter 3.04 Container Coating with 8-2 TBA Solution SsteevreirlaizledpoulnydperropUyVlelnieghgtalolvoenrnjiugghstwiintha Bliidosh(oPordt.iuFmivpeadcakyasgibnegf,oCreasteheIiDnFitYiaZt2ioQn0o7)tfhwetreest, (onMeeogfohthme-jcumgs=wa1s7.5f)i.lleAd w0i.4tmh4Lliatelrisquooftsotefri8l-e2wTatBeAr fsrtoocmkBsaorlmusttioenad(3E-mPgu/rem.syisnteetmhanol) wrianssiandgdweidthtosttehreiljeuwgfaotrer,aftihneajlucgowncaesntcraaptpieodn foofr3l0a0tergu/sLe taondmawkaes8t-i2rTedBfAorsa4tudraayst.ed mAifneerral medium. 315 PreparationofSaturated 8-2 TBA Solution FfirvoemdBaaymsstbeefaodreEt-hPeurienistiyasttieonmo(Mtfhegothetm,-acnmoth=er17j.u5g) waansd wfialsledauwtiotchla3velidt.ersAfoifsetrecroiolleinwga.ter dToBwAnI,L0.a6n0d4t2hegsooflu8t-i2onTwBaAswsatisrraedddoendatosthiepljautgeffoorra4fdianyasl.coTncheentsroaltutiioononfin2t0h1ejmugg 8w-a2s "tTrhaenssfuerpreemdattoanstterwialescternatnrsiffeurgreedbattottlhees apnred-wcoaastecdenjturgifpurgeepdarteodrienmSoevcetitohne 3pa.r1t.i4caulnadrwmaatster. referredassaturated 8-2 TBA solution. 3.16 Preparation of8-2 TBA Saturated Mineral Medium TThBrAeesmoillultiiliotner(3ealictherosf) aMnidnewraas smtioxcekdsoblyutiinovnesrsAe,lyBs.hCa,kianngdtDhewjaugs.adTdheedm1i0xtehde ssoaltuutriaotnedw8a-s2 refered as 8-2 TBA saturated mineral medium and was used the next day 3.07 Activated Sludge Collection MAupnpircoixpiamlatWealsyte4 TlrterasotmfeancttiFvacaitleidtysl(uPdgOeTwWa)s-cAoelrlieacttieodnfBraosminth#e2CiotnyDoefcWeimlbmeirng3t1o,n20(0D2E.) Awaesrmiarxreidvibnyg abtritehfelyessthalkaibn,gthtehesljuudggteowsaussapsesnidgtnheedmaincrIoDornguamnbisemrs.E93A3ft8e6r-s1e0t5l.inTghtehesludge sludge for approximately 15 min to remove coarse matters, the upper aqueous phaseof the EMSER13-03/4842 Page 8or28 000012 DuPont EMSE Report No. 13-03 sacltuidvgaetewdasslfudegreefdlrtahtreo.ugAh 2a0NmylLonalnieqtuowtiotfh tahpofrlersaitzeeowfas85truamn.sfeTrheidstoiasgrlefaesrrsedsc1in0tailltahteion vrieaflerwrietdhtao apslaksitlilcetdrsanlsufdegrepfiipletrtattee.and was autoclaved. The autoclaved sludge filtrate was 318 Test Units Tcersitmpvecsaspesl.s wTehreep6r0e-mclLeabnoirnogsiwlaiscadteongelabssy sreinrsuimngbotthteleasluwimtihnupmre-focilleaanneddsaelputmaiwniutmh-mleitnheadnol once and then with sterile deionizedwate tree times. 32 Test Conduct Fanoduratlyupmeisonufmexfpoiel.rimTehnetaplret-rleeaatnmeinntgswwaesredocnoendbuyctreidnswiingththperem-cwlietahnemdetghlaansoslwaorneceanadndsetphtean with sterile deionizedwater3 times 321 Treament]-8:2T84 ral Medium sivated Sludge Filrate 8F-o2raTBotAalsaotfur1a6tegdlamsisnserearlummebadtitulems a(4ndre0p.l1i7ca5temsLxo4fascatmipvlaitnegd tsilmuedgpeoifniils)r,ae34w.a6s2amdLdeodfto epraec-hocifeatnheedgallasusmisneruummfobiolttalneds wPiTtFhEsplialsitciconteranssefpetrapipettes. The bottles were crimped with 322 Treatme2n-t 8-2 TBA Saturate i il 8F-o2raTBtoAtalsaotfur1a6tegdlamsisnserearlummebodtitulems a(4ndre0p.l1ic7a5temsLxof4ksialmlepdlisnlgutdigmeefpilotirnaitse)w,a3s4.a8d2demdLtoofcach ocfletahenegdlaaslsusmeirnuumm bfootiltlaensdwiPtThFEp!lsasitliicctornaenssfeeptrap. ipTehtiess.trTehatemebontttsleesrvweedreascarnimapbieodtiwcitchontprreo-l. 323 Treatment3- Mi i Filtrate for8-2 TBA Spike RFeocroavteotrayl of 16 glass serum botles (4 replicates x 4 sampling time points), 34.82 mL of the smeinreurmalbomtetldeisuwmitahnpdla0s.t1ic75trmanLsfoefr apcitpievtaets.ed sTlhuedgbeoftlrsatweerweascraidmdpeeddtoweiathchproef-ctlheeagnleadss aluminum foil and PTFEsilicone septa. `1A4t0eaacLhosfam8-p2liTnBgAtismteopcokinsotlsu(tDioanys(101,2.7,5 1m4g,/aLndin28e)t,ha4noblo)ttfloers wefirnealspcioknceedn(tdroasteido)n woifth 4wi4t8huagn/Lo.rbiTthoerybosthtalkeesrw(eNreewinBcruubnatsewdicfkorSacpipenrtoixfiimcaCtoemlypa3n0ym,iMnowdietlh G2-001-0,30Se0riRaPl Mnusmhbaekring 880511137) before sample collection and sample extraction 324 FTorreaattotmaelno4tf8-gMlianssersaerluMmedbioutmsp(l2urseAplcitciavtaestexd4SlsuamdpgleiFniglttirmaetpeoinis), 34.82 mL of msienreurmalbomteldeisuwmitahnpdla0s.t1i7c5trmanLsfoefarpcitpievtatteesd. sTlhuedgbeotftilletsrawteerweascraidmdpeeddtwoietahcphroef-tchleeangleadss aluminum foil and PTFE/siicone septa. These samples served as sample mairix controls during quantification of 8-2 TBA and is possible transformation products. EMSERI3-03/4842 Page 9128 000013 DuPont EMSE Report No. 13-03 325 Test Conditions andSampling 3251 Sample Incubation o"Trhbeitcorriymspheadkegrla(sNseswerBurmunbsowtiecskwSecrieentiinfciucbCatoemdpwaintyh,2M0o0d-e30l0GR1P0,MoSfesrihaalkniunmgbewirth an t8e8s0t5s1y1s1t3e7m)waatsrmooonmittoermepderaantdurreecionrtdheeddtahrrkoiungshioduetatchehecmoiucraslehoofotdh.e sTthudey.temperatureofthe 3252 Sampling Interval Four crimped serum bottle for Treatment 1-3 and 2 crimped serum botles for Treatment 4 wDeecr-e2s00a2m)p,leddayfo7r e(x7t-rJaacnt-i2o0n0o3f),8-d2ayTB14A,(1f4l-uJoarni-d2e0(0F3~)i,ona)n,dadnadyo2t8he(r28m-eJtaanb-o2l0i0t3e)s.at day 0 (313253 Sample Storage Analytical samples werestoredatapproximately -20C. 33 Sample ExtractionandAnalysis 33.1 SCampolellecantdExitraoctinon 33.01 Sample Collection bAottdtlaeyssw0e,r7e,t1u4r,neadndup2s8i,dcerdiomwpne.d sAamfpolaelobfo1t0smwLeorfe trhemtoevsetdmefrdoimumthfersohmakeearchaonfdtthhee b2o.t3t0lemsLwagslawssitshedrruamwnbwotittlehaand10c-omveLrpeodlwyiptrhoppyrlee-ncelesaynrcidngael.umFiivneummilfoliilliatnerdwwaassifnojercetdedininato fproeleyzperrofpoyrlefnurethteurbeantahlaytsicso.naAinnoetdhe0r.055mmLLloeffinNthseodsyiruimngheydwraosxiidnejefcotredfliunotroiade1e5xtmrLac.tion. 3312 Sample Extraction NFlauOorHidweeerxetriancctuiboan:tedThateo1o5 mmLtemppeoraltuyrepforrot3u-bp4eshycowlnittehai2nn5i0neg 3h0e0$RmPiM.otefstsmhaekdiingu.m Tplhuesn 0s.a0m4p2lemtLuboesf6wNerHe,sStOo,rewdaisn aa-dd2e0dCtefarecehzeorftfhoreltatuebresfltuoornieduetraanlailzyesitsh test medium. The 8a-d2diTnBgA5.e5x6trmacLtoiofn:6NBeHfo,rSeOexitnra5c0ti0onm,LMoTfBcEh-ilHl,edSOM,TBsEo.lveAnt 3s0ysmtLemawliaqsuoptroefptahreedcbhyilled MteTstBmEe-dHi;uSmO,wawsaswitthhednrianwjnecftredomintthoe ebaoctthleosf.tAhRcerritmhpeedMTsaBmEp-leHbyoSttOl4eswaafserintjehcete1d0, mthLe of s2a5m0p-l3e0b0oRtPsMwsehraekiinngc.ubAatfeedrsaftrionogmttheempMerTatBuErepfhoarsea,ptphreoxcirmiamtepleyd 2sahmpwliethbottles were dpiepceatptepetdo a5n0dmtLhepMolTyBprEoppyhlaesneefcreontmreifaucghoe ftutbhees saanmdpkleeptboitntlaesfrweeazsert.raAns2fe0rrmeLd wailtihqauotgloafss MpoTlyBpErowpaylseandedceednttroitfuhgeestaemspl.e bTohreessamafptlretbhoettfliersstwMerTeBrEecpahpapseedwwaisthttrhaensofreirgriendalto the a1lhumwiintuhm2-5li0n--ed30T0efRloPnMd ssheapktianag.ndTwheereMiTnBcuEbaptheadsaetwoaosmthteenmtprearnastfuerrerefdorwiatphparogxliamsastpeilpyette M10TtBheEpoplhyapsreowpaylsecneencternitfruigfeudgaettaupbpersotxhimaactoenltyai2n0e0d0thRePfMirsftoMr T10BmEinp.hasAe.20TmheLcaolmibquionteodf t2h0emMLTBglEassphsacisnetiflrloatmioenacvihalofwtihteh cefnotirli-fluigneedtucabpe.s wFarsomtraancsfheorfredthewigtlhaassglscaisnstiplilpaettitoentvoiaals, EMSER13-03/4842 Page 100728 000014 DuPont EMSE Report No. 13-03 2ClmeaLneadliaqluuomtoinfutmh-eliMnTedBcEripmhpasceapwafosrtGraCn/sfMeSrreqduatnot.iafGicCativioanloafn8d-2waTsBsAe.aled with pre33.13 Temperature Measurements `The temperatureofthe test system was monitored and recorded throughout the course ofthe study. 332 Analytical Methods for Test Substance and Products 33.21 Analysisof Test Substance 8-2 TBA Analysis: Analytical standards: "PTrhoeduIcHt,s,1HW,e2sHt,C2oHl-upmebrifal,uoSrCo)dewcaansu-1s-eodla8s-t2heTBanAa,iytCiAcaSlHst6a7n8d-ar3d9.-7T,h9e7.16D%,,1ODs,k2wDo,od2.D, S3.t1o5cCk-shoelputtaidoencsa(f1lu0o0r0omdge/cLa)noolf(tDh-e8a-n2alTyBtiAc,alDsutPaonndtar)dwaansdutsheedianstetmhaelisnttearnndaalrdstwanedraerd. eaprcehpcaarleidbrinatmieotnhainnoMlTaBndErbeyfrdigielruattieodno. fTthhee cfarleisbhrlaytimoandseta5n0da-rmdgs/wLesrteocpkrsepianrmeedthfarneoslh,ly for CTyopnisctaalnlty,fetvheelocfaliibnrtaetrinoanl ssttaannddaarrddswwaesruesmeda:de~3in00thuegr/aLnogfeoDf-28-52-1T0B0A0.g/ThLe8c-a2liTbBrAat.ion c(uDr-v8e-s2wTeBrAe)coannsdtrtuhcetreadtuoosfintghtehecornatcieoonifratthieonpseoafk a8-r2eaTfBorAioanndm/tzhe3i1nt(e8r-n2alTsBtAan)daarndd. vAzn 33 oefxsaammplpeleo(faE9c3a3l8ib4rEa1t-io2n9)cusrevpearwaatisogniwvaesn ignivFeinguireFAi-gu2reanAd-3a.n example ofa chromatograph gAla0.s5 s mGLCavliiaqlu(o1t.o7 fmLthevMolTuBmEe),ph3aisLeforfo5m0 tphge/GmCLovfiDal-s8(-S2ecTtiBoAn 3i.n3t.e1m.a2l) wstaasndpalradcewdaisnaadded w10asthaenvailaylzuedstwiaicnGeCgbysayrGinCg/e,MStheinvsiatlrwumaesntcaapcpcoerddianngdtso tuhebfjoltleoowcainnatglycesoindsd.itioEnasc:h sample GC/MS system: HDePte6c8t9or0 (PAlguisleGntC),(AMgPiSle2n-tM)u,lHtPiP5u9r7p3osMeaSsasmpSleelerctive (Gerstal) Column: 'DB-5MS, 30 m x 0.25 mm, | wm film (Agilent) Temp. camp: Initial: 80C for 2 min 20Cimin to 120C 50C/min to 300C and hold for 3 min Flow rate: 1.0 mLmin; He; constant flow mode split 51 Inlet temp.: 250 Injection volume: 20 MSD transfer line temp.: 280 Tonization: ELT0eV SIM ions monitored: mlz: 31,33, 95, 98; dwell time: 25 ms for each ion EMSER13-03/4842 Page 11 0f 28 000015 DuPont EMSE Report No. 13-03 Retention time: 82 Telomer B Alcohol (8:2 TBA): 497 min Intemal standard (D-8-2 TBA): 495 min Fluoride Analysis: Analytical standard: Ccaelritbirfaiteidons.taTndhaerdcaolfib1r0at0imong/stLaonfdafrldusorwiedreeinmawdateern(tThherramnogeOroif5on~) w1a00s pugs/eLd fbyodsitlauntdiaorndof h"Tehe1r0m0oOmrgi/oLn)oafnfdluodreiidoensitzaenddwaradtwrifthroTmIBSaAmBsIiIead(oEt-aPluorneiscyssttreemng(tMheagdojuhsmt-mcenmt b=uf1f6e-r I18l). HfaolrfthsetdrielnugttihonT.ISAABImIL (aolinqeuoptaortfofcaTcIhSoAfBtIheIsapnldusarodnesoplaurttioofndweaisonuiszeeddfwoartfelru)orwiadse used analysis to consiruacsttandardcurve for quantification of fluoride in samples Quanifcaion of Fluoride: "pTohleyp$rmopLylteesntemteudbeisuwmtahsatthwaawsedtraetaoteod mwittehmNpearOatHuraenadnHdiwSaOs4cfernoimreiafucghedo.ftFhoeur15mimLler `niLqouoftTfIrSoAmBeIaIch osoltufhtiont.ubeAsftwearsmtirxainnsgf,ertrheedm10e.d2i0ummwi.aspoalnyaplryozpeydlfeonreftluuboeritdheatuscionngtaai7ne1d04 e'AlePcltursodpeH(ITShEermmeoteOrri(oTnh,emromdoeOlr9i6o.n0,9s,erlioatl##G0X616)8.14A)faenrdfaillionngpltuhse rfleufoerriedneceseclhecatmibveer with rtheefesraemnpceleelmeectdrioudem fainlldinfglusoorliudteiosnt(anTdhaerrdmsoolOurtiioonn),antdhetheeleccotnrdoudcetiwvaistyinisnermtieldliivonlotseawcahsof r`egcenoerrdaetdeadfbeyrpSlomttiinngofthienscutbaantdiaornd. flEuaocrihdesacmopncleentwraastimoenasinurLeOdGtwsiccea.le Aversstuasndmairldlicvuorlvtse was (Figure A-1) for quantificationof fluoride. 40 RESULTSAND DISCUSSION 4.1 UTanbdleerAtDhe).est conditions, 82 TBA is no readily biodegradable (Figure 1, Table 1,and + 9Sp1i2ke3r%ecaotvdearyyo7f, 984-2+T6BAtfrdoamyt1h4e,taesntdm7e4d+iu5m%aavterdaagye2d81(0T6a+bl5e%A-t2),daiynd0i.cating mheadtiMumT,BeExceexpttraoctriodnayme2t8hsoadmopblteasined a good recoveryof 8-2 TBA from the test Acotmapyar7e,dtwhietlhosasboiofti8c-2coTnBtrAolivnetsessetlsve(sTsreelast(mTernetat2)m.ent 1) was ~8% more(p<0.05) + (A9t<d0a.y011)4c,otmhpealrosesdowift.h2 aTbiBoAticincotnesttrovlesvseeslssel(sTr(eTartemaetnmetn1t)2w.as ~13% more AStysatyem 2a8n,dtthheecoanbcioetnitcractonitornoslso.f 8Mo2sTtBkAelw,erheenloosdsisotfin8g-u2iTshBabAledubertiwneethnisth2e8-tedsaty b{eisottwraanssfdourema1t0iaobniotic removal mechanisms ~ volatility and/or adsorption athr than Inltoshseisapboisostiibclcyonctaruosles,d tbhy vcoolnatcielniizrataitoonnodufr8i-n2g tThBe Aincduebcarteiaosnedorcobnytiinnucooumslpyl.eteThe E EMSERI3-R 03/4842eee eee Page 12028 000016 DuPont EMSE Report No. 13-03 extraction due to adsorption to the glass wallsof the test vessels and/or the sludge inoculum. 4.2 Under the test conditions, no defluorination of 8-2 TBA occurred during the 28-d test The fluoride concentration was the same in the test medium during the test period, averaged 17.9 + 2.6 pg/L at day0, 17.1 08 ug/Lat day 7, 17.4+ 1.4 pg/Lat day 14,and 175+ 1.0 pg/Lat day 28 (Table A-3). Under the test conditions, perfluorooctanoic acid (CAS# 335-67-1) is unlikely to be a `major metabolite because defluorination was not observed during the test. On a `molar basis, for one mole of 8-2 TBA to be converted to one mole of perfluorooctanoic acid, two molesoffluoride would be released. No increase in fluoride concentration was observed during the test. As a result, the absence of fluoride concentration increase during the test indicated that perfluorooctanoic acid is unlikely a major metabolite (>10%of otal mass present) of 8-2 TBA `biotransformation under these test conditions. 5.0 (CONCLUSION Under the test conditions at room temperature (~22C), 8-2 TBA is not readily biodegradable. Confirmed expectations that abiotic removal mechanisms -- volatility and/or adsorption will be factors that need consideration when designing future studies with objective of achieving mass balance. 6.0 LIMITATIONS OF THE TEST Due to the low water solubility, volatility, and strong surface adsorption properties of 82 TBA, its difficult to conducta test with a good mass balance (90%) for the abiotic control, whereas, the mass balance achieved in this study was only 67%. During the test, the concentration of 8-2 TBA continued to decrease in the abiotic controls, complicating the experimental data interpretation. `Taken together, the conclusion drawn from this testis indicative, not conclusive. Further studies willbe conductedtodetermine the potential major biotransformation products of 82TBA. 7.0 REFERENCE 7.1 OECD Guideline for Testingof Chemicals, Section 3: Ready Biodegradability: 301 D. Closed Botle Test. (1992). EMSER13-03/4842 Page 130f28 000017 DuPont EMSE Report No. 13-03 FIGURE 1 8-2 TELOMER B ALCOHOL (8-2 TBA) CONCENTRATION DURING THE 28-DAY TEST ! i.23Ed :< 2 i3g > Daysafterthe initiation EMSER13-03/4842 Page 14 of 28 000018 DuPont EMSE Report No. 13-03 FIGURE 2 FLUORIDE CONCENTRATION DURING THE 28-DAY TEST* "Zeo-- AKcitlievdatseldudsgieudigreatfietr+at8e-+28T:B2ATsBaAlusraattuerdatmeidnemrianlermaeldmieudmium = =| 0 by3 2 fFR 3[ 5 os ww mw Days after the initiation + Trhaephteiss wdearsivienditifaotemd otnhe3o1riDgeincaelmdbaetra 2of00T2abalnedAf-i3.nished on 28 January 2003. The EMSER13-03/4842 Page 15 of 28 000019 DuPont EMSE Report No. 13-03 TABLE 1 CONCENTRATION OF 8-2 TBA AT DAY 0 (31-DEC-2002), DAY 7 (7-JAN-2003), DAY 14 (14-JAN-2003), AND DAY 28 (28-JAN-2003)* Trpeot Trostments Time. oor sTruopaeomMemnt1e-pAcntBua2tTedEA 70 Sanat mire magn Wo one dtowaxe wTHen2z00m8 maeaw Bata Concentraton we wfeist eseno wan TestSubstance remaining gaCyom7pdaaryed14t,oDaanydd0a2y28 x 00 n TratmKaondtsa2g.e 0 310ncae 427 0 BSaarsaiepdsie2rTBmA om 7 wo Tan tena wise sso a " Bm suman ess o + "Thistable is derived from the original dataof Table A-1. +4 TMeesatnsubssttaanncdearrdedmeaviinaitniogna:tnD=a4y.7s, 14, and 28 compared to Day 0,%= (C:/Co) x 100, where: Cy=8-2 C=82 TBA TBA concentration concentration at at Day 0, Days 7, I4,and 28. EMSER13-03/4842 Page 160128 000020 DuPont EMSE Report No. 13-03 TABLE 2 DAILY TEMPERATURE READINGS WITH A CALIBRATED DIGITAL THERMOMETER INSIDAE CHEMICAL HOOD WHERE THE TEST VESSELS WERE INCUBATED Time. E3 1 2 3 . 5 5 7 10 " 2 8 " 1s " 1 1 2 2 2 a x Average, n= 20 `Standard Deviation oats 310002002 14a 2003 24a 2003 an2003 an2003 S023 un2003 72003 Suan2003 San2003 104002003 11402003 124002003 13402003 144an2003 164an2003 16-Jan-2003 174202003 184802003 169302003 20402003 2102003 2402003 20am2003 2a2008 254an2003 28-Jan-2003 27-4an2003 28402003 Temperature 28 2281 28 a 2st 2st 24 23 29 21 20 28 2a8t 20 22 20 23 29 201 201 29 21 21 23 29 2501 231 a7 20 20 [x] + Temperature reading in the test lab recorded by a calibrated Dickson rreecaodridnegrs (bMyotdheelrTecHoDrXde,rsweerirael nnoutmubseerd0f1o1r1c8a-l2c4u7l)at.ioTnohfettheempedraailtyure: average temperature. EMSER13-03/4842 Page 170f28 000021 APPENDIX A: DuPont EMSE Report No. 13-03 EMSER13-03/4842 Page 18028 000022 DuPont EMSE Report No. 13-03 TANAABLLYETAI-C1A.L RESULTS OF 8-2 TBA CONCENTRATION AT DAY 0 (31-DEC-2002), DAY 7 (7-JAN-2003), DAY 14 (14-JAN-2003), AND DAY 28 (28~JAN-2003) m no Teens d me chiWsyitAes e Am`vwiAgoen cEoncTamesAtont wo ot Contec Tomause asc5heTon 01 nt 0s oT mn amsasz uplmnnn 0 20 womm wo on emia oviwwm Ps Po eames 4smow wr a Averages Sanda Dviations Ow oss MESSTrameanni2zTeedxcgume 1 o2o on we emacs 0 2: wmer oe wo emer 0 aw @ 0 sem eamesa Average: sandarsDoviton 0 o Cw w Is m 027 COIMELS TfTarameesrnsActoanegs |7 1 9o9e9 0s " [rer TaFE ome ns wr emacs Ta5 emwe ws " comme J---- ToeCmme se " uso CRIMESTm ammae Taloasnsniae T1Tom984 emis Tz ws uz ws wo 1 camnacsan PEEPw Ir 0s mein soe omme ws - Average:Standard Deviation vm wae EMSER13-03/4842 Page 190 28 000023 DuPont EMSE Report No. 13-03 TABLE A-1 (CONTINUED) ne Teroest NUE ranches cones wmmeedmsmenpsaasmzetTuInA emma snes Average Standard wvation EONMEL ramantz totsge foro ps 82TBA comea Ese mesmenimetn eneesss esmesse Ww Twa cm oAnncaenyviA satlon 7 ED HT 1 185 87 14 2 2 783 om wos me PE woo me ms 1 98 wr 14 2 ga a wos me a ws woo wo AAvvmaeiRsl i or n a ns sas os 0 on Average SancadDeviation com RUSTraiment 1 Acres BTS 5 Comaacsseswuemmmaepousaa0i2mTeesAn 28 21mTea ne zm mess Boa es ese a ee smi Aversa SandadDeviation wa as os om CONES! ramet kmasisge BT 82 "2 ropa 8278 Conesesume mosanetn 23 Te 2 919 we 2 ar eos wa se = a wes Eonmson wa ee 9 PE Average StandardOwain C+F.i=cn8a:2oTBnA acnalyeticoafln6av2earTaBgtAe,vCaloyue,=Cnx (Versa Vo where: VV+,Srt2as5nd=amr5dL0Dm(eTvLeisat(timToeBndEiwuamsuscuesdae1eldxlfieordxatucasichneigohense) fmineadlicuomn)c.entrationof3-2 TBAT = 4 econacanimatont Ch 0 1 ww 1. wen " 0 ES "2 meas 0 0 " " ws Ea 0 een -- EMSER13-03/4842 Page 200 28 000024 DuPont EMSE Report No. 13-03 TBLEA-2. ANALYTICAL RESULTS OF SPIKE RECOVERY OF 8-2 TBA FROM THE SAMPLE MATRIX (TREATMENT 3, DAY 0 (31-DEC-2002), DAACTYI7VA(T7E-DJASNL-U20D0G3E),FIDLATYRA1T4E P(1L4U~SJMAINN-2E0R0A3L)M,EADNIUDM) AT DAY 28 (28-JAN-2003) " TTT Time Rep G5oam1cvn8tiiAtson_ BAv2vaTatBeAn FCionntea8miinTiBoAnt %oRlocSopmkyte EmuE ms 0 1 om wr Pa on rr om = PA ws ems 0 D 3 aom = PA wo 3m [rE = mn or Average:StandardOevitP on E ex [Et Vm PA " o wus 1:pomom we " w [ee 3m wo " " women 7 4m mn PA w Arnage SantanaOoiation womes wo 1 an m a nes " asses wo: aomm mn " ws mse Ww 3mom xe w " [Er 3m m PA us AveagesSanta oaton sz EMSER13-03/4842 Page 21 of 28 000025 DuPont EMSE Report No. 13-03 TABLE A-2 (CONTINUED) --_-- No Time ep a5naIlyTtBicAal AmaSiAyTtBcAal Concentration _ Average E22 we Wr eonmesss 28 1 1 182 . 100 eonmEss Bz 108 168 2 168 eames 2 3 164 n eonmesss 2 34 1i8"1 18 Average 2 Standard Deviat+ion 184 + Final 8-2 TBA concentration, Cy = C, x (Varae/V) where: C= 8:2 TBA analytical average value, Varu=e 50 mL (MTBE used to extractthe test medium). V,=25 mL (Test medium usedfro extraction). 3of% spike recovery, R = (C//C) = 100, where: C= final 8:2 TBA concentration, C,= Concentration of 8-2 TBA spiked into the sample bottles (448 CoFnRcweinteaitTiBoAnt mWTr 24 3% 2 2s pg/L) Re%cooivSemrhye? n " i J nz - EMSER13-03/4842 Page 220128 000026 DuPont EMSE Report No. 13-03 TABLEA-3. ANALYTICAL RESULTS OF FLUORIDE CONCENTRATION AT DAY 0 (31-DEC-2002), DAY 7 (7~JAN-2003), DAY 14 (14-JAN-2003), AND DAY 28 (28~JAN-2003) No Trmosomeent Tine ep cFoSnnecaaiwnetailon FAAvvleayriaocgaels Fcuooncnaanierfadoomnt -- EOMUEN rosment1 Acted ww0 1 w1r07 w10r8 wnrz sige ale plus 62TOA Ts orsaues2 sedmncoimedm 0 2 93 oe 10 2 oe comes 0 3m 9 158 se eomssaess 0 4s 158 Avarge Standard vitor: a nssze ESAELS rraaument2- oadsicge 1 98 01 102 rae pos 82 TBA 1 oases nmesmosaimedm 0 2 as 89 os wz 2 m2 esses 05 a a 02 som esssaess 0 a re 158 on Average Standard Deviation seas EOAELS rresumant1- Acres TT a we supefaepus 62 TEA s " Cosas SUITOR, gy a 102 osm PE Ts nm 8s 0 a es Esme Tee ar 102 Average Standard Devin ee nazos CUMENS rgmentz-masiige TT 100 mn ne feat pn 82 TEA ' 7s Essai uumamneninedn 7 2 8 0 60 x nz eonmesa Ta as on 102 as mes Tew "7 56 Average Standard Govition 1eazos EMTSEERE1T3-0E5/P4842eee eee Page 23 0128 000027 DuPont EMSE Report No. 13-03 TABLE A-3 (CONTINUED) No Tmrvsiwment Fe Te fcoAnoncaeymmiicmatlon FAAonvaneyidnceasl Fucoonicadntsrtatioint -- CONESraaument1-Actus #1 & wir awer m1l08 sudge fate pis 62 TOA ' 7 EonmuEs0 sumedmneimedm 14 2 83 a 02 ze omsaesst w oa ee os 0 a ee comes woe or 104 Average StandardDeviton es asia EORUEIDrrsetment2-Ktadsicge M1 os os 0 te pus 2 To ' Conespumameeaineton 14 2 a 78 a 0 ze Ee ws ow a 102 sm eommesss wom a 02 ow Average+ Standard Deviation ESIMELST rreagment-Acoed 21 0 16az0s 03 186 Sucge sepus 52 T8A 1 03 onset dummmewsmetin 73 2 88 90 0 omens PR 3 a0 82 04 Eomiesen se moa 8 os 0 Average Standard Govition PE 5210 CEI ramen losocge sie pus 82734 BT ' 83 oe 90 100 Eonsierer Huamoasnen 2 2 8s 0 1s eomieses 2 wo 3 es or Ex Er a ee oa 10 os nz Average+ Standard Deviation PE szer +=FinFalluNouroidreidaenacloyntciecnatlraatvieorna,geCv=alueC. x[(V.+V, VJ where: VVS,t=8an8dammriLd D((TeTevIsiStaAtmiBeoIdnIiwuabmsufucfsaeelrdcuafloadrteuddtoourestiiddhneg tmehseetasimnueardleimfeulnumo)r.oirdeucoornciednetrmaetaiosnusrenm=en4t. ). -E--MSER13-03/4842 Page 24 028 000028 DuPont EMSE Report No. 13-03 TABLE A-4. PREPARATION OF THE MINERAL MEDIUM FOR THE TEST - the following stock tical ot OECD Guidelines) (@ Potassium dihydrogen orthophosphate, KH: PO, Dipotassium hydrogen orthophosphate, Ks HPOX ADimsmoodniuimumhycdhrloogreidne,orNthHoCphIosphate, NasHPOX Dissolve in water and make up to 1 liter. The pHofth solution should be 7.4. 301 D Test 850g 21758 2839 030g (8) Calcium chloride, anhydrous, CaCly 27.508 or Calcium chloride ditydrate, CaCl 2H2 0 36408 Dissolve in water and make up o 1 ltr. () Magnesium sulfate heptahydrate, MgSO.TH: 2508 Dissolve in water and make up 101ltr. (@ tron (1 chloride hexahydrate, FeCly6H:0 025g Dissolve in water and make up to1 lier (6) Filter each stock solution througha sterilized filtration unt with 0.2 um size pore. aNdodt:onIendorrodpeorftcooanvcoeindtrhaatveidngHtCo ppreerpalirteerr. onIf(a11pr)ecchilpiotraitdeefsotromcsk sionlutsitoonckimsmoeldutiiaotne,lryepbleafcoeretuwsiet.h a fresh-made solution. a EMSER13-03/4842 ee Page 250128 000029 DuPont EMSE Report No. 13-03 AFIFGLUUROERAI-D1E STANDARD CALIBRATION CURVE USED FOR FLUORIDE QUANTIFICATION OF SAMPLES ES3384EJ-1 TO E93384E-64 2 ow Y= 4g016 001(9R%1=0386) 2 a5 oe So 2gsSnw T os ~o os EE EE millivolts A REE enea eeen EE EMSER13-03/4842 Page 26of28 000030 DuPont EMSE Report No. 13-03 FIGURE A-2 AE9C3A3L8I4BERJA-TI1OTNoCEU9RV3E38U4SEEJD-F2O8R 8-2 TBA QUANTIFICATION OF SAMPLES Taw |Response Ratio ] , || | as / | // / yd | | i He os || - - -- || P Resoy macio +A3.3C 10001 + r duc <3 t 470-000 iAntcealmiablrasttiaonndacrudr:ve3u08sgdfLo osfaDm-p8l-e2s TEB9A3.384ThEeJ-r1atnogeEo9f33co8n4ceEnJt-r2a8t.ioTnhsoefco8n-c2enTtBrAat:ionofthe 25.5 1020 gL. EMSER13-03/4842 Page 27 of 28 000031 DuPont EMSE Report No. 13-03 FAICGUHRREOAM-A3TOGRAPH OF SAMPLE E93384E J-29 USED FOR 8-2 TBA ANALYSIS Abundance a000 00 | 4000 on 31.0000.7010 31.70) 012203230 4.97 = 10745l5l 0 55 60 65 70 75 80 85 90 9S 5 10e 0 ime--> [Abundance a0 495 lon 3003270103270)012203230 6000 | wn| 2000 | 0 70 45 50 55 60 65 70 75 80 85 50 95 100 Time--> Sample E93384 EJ-29; lon 31, retention time 4.97 min = 8-2 TBA; lon 33, RT4.95 min = D-8-2 TBA (internal standard). TTTTTIE--------------------------------Tr EMSER13-03/4842 Page 28 of 28 00003%