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ARaz6-- 3303 ~D [Swd| yNmoai0.1] ARAE - 3363 Study Report Volume 31 a i (fon-GLP study) October 16, 2003 Chemicals Evaluation and Research Institute, Japan Kurume Laboratory `Company Sanftized. Doss not contain TSCA CBI Tables L Table 1. Table for calculating percentage biodegradation by BOD Figures Fig. 1 Chart ofBOD Fig. 2 Mass fragmentograms of LC-MS analysis for calibration curve, and calibration curve Fig. 3 Mass spectra oftest substances ja) Study No. 14101 2 Company Saniized. Doss not contain TSCACB ~) Table of Contents COMISSONEA BY. TESHOG FACHLEY cvssmh J SCBA. A APPIOVal Of SHAY REPOTLcsosennnemsssssmsssesssssnd 1. TOS SUBSIANCE rvs ~~) 2. ACHVaIEd SIGE corinne] 3. CONUCHING the STAYvrs 4. Confirmingthe Study COMONS crv l2 =] Study No. 14101 3 Company Sanitized. Does not contain TSCA CBT i) Title: Ssttuuddyy) Asin by Microorganisms (non-GLP Commissioned by: DuPont KK 1-8-1 Shimo-meguro, Meguro-ku, Tokyo 153-0064 Testing Facility: Kurume Laboratory C19h-e1m4icCahluso-Emvaalcuhait,ioKnuraunmde-Rsehseia,rFcuhkuInosktaitPuree,fJeacptaurne(CE3R0I-)0023 Purposeof Study: Tbyo mdiectreoromrignaentihsemsd.egree to ic ERY degradable Procedures: `Tphrioscesdtuurdeys.was carried out in accordance with the following test (1) i"nTeTsetsitnMgetthheoDdesgrfaodraNbielwitCyohefmCihceamlisc(aNlostibfyicMatiicornoNoor.ga5noifstmhs"e PNoltainfniicnatgiaonndNCoo.o6r1di5noaftitohne BPuhraeramua,ceEuntviicarlonAmfefnaitrsAgBeunrccya;n, MthieniBsatsriycofInHdeuastlrtihesaBnudrWeealuf,aMrien;is1t9r7y4oNfIontitfeimcaattiioonnaNlo.Tr3a9d3eof ~) (2) "aRnedaIdnyduBsitordye;gdraatdaebdiJluiltyy:13M,od19i7f4i)ed MITI Test (I) (Guideline 301C, July 17,1992)," in OECD Guidelinefor Testing of Chemicals `oCxaylgceulnatdieomnasnodf.biodegradability were carried out using only Schedule: Study began on: TSetsutdiyncgopmeprlieodt:ed on: April 30, 2003 OJcultyob2e,r20160,32t0o0J3uly 30, 2003 Personnel: Coordinator: Researchers: T`eTsotmionhgiSkeocYtiOoSnH1IDA" Tomohiko YOSHIDA MYaassaunkooriMATATRSUUKNIOBE Approval of Study Report: [signed] Tomohiko YOSHIDA Study Coordinator October 16, 2003 ~~ * rsssor'sNoe: Spelingsofpesos misray ned beverific StudyNNeo. 14101 4 Company Sanitzad. Dos nt conteln TSCA i ~ ~~) Summary TiotfSltuedy StudyofSD cscnion by Microorganisms (non-GLP study) Stdy Conditions (1) @) ACcotnicveantterdatsiloundogfetecosntcesnutbrsattainocne:: 100 mg/L. 30=o.(I (3) Amount of test liquid: 300mL (4) Test liquid incubation temperature: 251C (5) Test liquid incubation time: 28 days Measurement `The biochemical demand analyzer. oxygen demand (BOD) was measured usinga closed oxygen TestResuls Percent degradation, based on BOD: 18%, 16%, 17% Average: 17% ~9 Study No. 14101 5 Company Sanitized. Does not contain TSCA CE! " 9 Nome 1. Test Substance Lot No. nN TPueristStyubstance:(SND ) ) ) Th est bse va gaars hdineptdy off) ~3 Study No. 14101 6 Company Sanitized. Doss not contain TSCA CET J SY 5 --~ 2. Activated Sludge (21.)1PPllaacceesos fanCdolTliemcteisoonf: SludgeCollection SludFgueshwiaksoRciovlleercTtredeaftrmoemntthePlfaonltl(oCwiitnygotfenSaplpapcoersotihnrHouogkhkoauitdJoa)pan. KNiatkaakhaamimaRiTvreerat(mCietnytoPflaInsthi(Cniotmyaokf iOnsMaikyaaigniOPsraekfeacPtruerfee)cture) HYiorsohsihniomRaiBvaery((CCiittyyofofTHoikruosshhiimmaa iinn HTiorkousshhiimmaaPPrreeffeeccttuurree)) FOcuhkiaasihiTbraeaTtrmeeanttmePnltanPtla(nSthi(nKjauskhui-kmua,-gTuonk,yIob)araki Prefecture) LSahkineaBniowRaiv(eOrts(uNiCiigtaytainCiSthyi,gNaiPirgeafteacPtruerfee)cture) Dokai Bay (CityofKitakyushu in Fukuoka Prefecture) `cWoansttaecwtaSwtaietmrhptlrbeeosatthtmatekhneetnspuflrarfnoatmsceswtoerfreetahmrese,wtaultranekerssalaunnddgdett.hheesatemaowseprheerteo.psSoailmapletshetwaakteenrf'rseodmge in (2) Time of Collection: March 2003 22m2ixPturIrenooofrdtehcreotafoebAmocavtsieinvtssaaltiuendidgtSehslneucuodgnlgilfeecotremditfyrooftmhvearaicotuisvsaitteeds swlausdgme,ix5elditweirsthof fliiltterrasteoffrom pfarideljftuirsalttteeerdf.rto0ma apcHtoivfat7e.d0sl1u.0d,getshtehnataewrearteedciunlttuhreecduflotruraibngotua3tnkmobnytphas.s"siTnahieg `tmhirxotuugreh waas *1l:iteArcstoifvaftiletdrastleufdrgoemoabtmaiixnteudrbeoyfctuhletuarbinogv,einslaucdcgoersdacnoclleewcittedh fsrecotmiovnar2i.o3usbeslitoews,, 10 s2u3peCmualtAtaeunrrtaitnaigcocnouonfttihnegfcoulrtaubroeuttanoknew-atshsitrdoopfptedhefotortaalboaumtou3n0tmiwnausterse,mfoovleldo.wiTngo wthhiiscwhas aadedraetdidonecwhalsoraignaaitnedcawrartieedr osuutch(faosraoblreiasntg3t0hemitnoutatlesv)o,laufmteeruwphitoch1050ltgr/sL. sNynetxhte,tic wwahdaidscethdewdtehacetheaibroorwivnaeasotpeaeddrdwaetaditoesnros1a0ws.e01r.01eswreeitp%te.haeteAcdcotnoicnvecanettreaadtdsialoyun.dogTfehsweyancstuhloetbtuitrcaiiwnngaestdtebemwypaectruaaltrtuuwrrieitnhwgianisntsheet 102542C. *2oc:fon7Ge.le0un4ct1or0sa.ei,opnespotfon5e0 agnLd. piontpausrsiifiuemd dwiahtyedr,roagnedn apdhjoussptheadtweitwherseoddiiusmsohyldvrteoodxirdeespteoc8tipveH Study No. 14101 7 Company Sanltized. Doss not contain TSCA cal ~ 3. Conducting the Study 3(1.)1 IMnPeiatrisaulreempenat orf SausftpoerintdheoedSnStouldsiyds in Activated Sludge: amounTthoefsaucstpiveantdeeddsslouldigdes tcoonbceenatdrdaetdi.on was measured inordertodeterminate the. Measurement Measurement method: date: ~~ MCeartrhioeddsoutSiunsapcecnodreddanScoeliwdist"h June 30,2003 "Plant (IS K Wastewater Test 0102-1998, section 14.1) Measurement results: OTfh4e,a6c0t0ivmagt/eLd.sludge had a suspended solids concentration (J2a)pParne)pPaurwraaitsfiioaenddodwfeadBttaeosra(alpmCriuoxldttuuucrreeedoMbfeyd3TimauLkma:esuagcihoPhfalrimqauciedustAi,caBl,; CThaendPhDardemsaccroipboeediianof s"ePcltainotnW2a1s.t)e5w0atasetrobTersitngMetthheotdotsal--vBiooclheuumimpcteaol 1Olxiytegre,annDdetmhaenpdH"w(aIsSaKdj0u1s0te2d-1t9o978.,0, (3) Reference Substance: dsuefgfriacdiAaentniitloilnneovefewlaanoisflauicsnteeidvaiaftsyttte7horecdaraeryfresyroaeunntdcetahsfetuebsrstut1da4yn.dcaeyTtsho,evaersreidffeeyrtteehnracmteitnvheaedlfusrelsoumdfgoterhhpeaesrBcOaenDt, `were setatmore than 40% and more than 65%, respectively. PS) Study No. 14101 8 Company Sanltzed. DossrotcontainTSCA CBY ~ 3.2 Prepaorf TaesttLiiqouinds 9 s`muebtsheoqSduseinxdtetlseycsritinvbesesedcstebileoslnoww3e..r3.eTphreosveidteesdt alinqdutidhsetweesrtelciuqlutiudrsewdeurnedperreptahreecdounsdiintgiotnhsegiven (1) (a) (AWdadtiteiro+notfestTessutbsStuabnsctea)nscyesatnedm Aniline: (one: test vessel [6]) fumisPhuerdifwiaesd wparetceirs(e3l0y0wmeiLg)hewdasouptloanceadnienlaetcetsrtonviecssbeall,antcheeann3d0madgdoefdtshoe2satmoplgei:ve a test substance concentration of 100 mg/L. (b) (Sl`uTdheg+ebasteasltcsuulbtsutraenmcee)dsiyusmte[misn a(nthraemeo:utnetstobvetsasienlesd[b2]y,s[u3b]traancdti[n4g])the amount of ofactthiveatseadmsplluedfguemaidsdheedd w(1a.s96pmreLc)isferlyomwe3i0g0hmedLo]uwtaosnpalnaceeldecitnroanticesbavlesasnecle, atnhdena3d0demdg 50.5 to give atestsubstanceconcentration of 100 mL. () (Sl`uTdheg+ebaasnaillciunel)tusryesmteemdi(ounme:[intatnvaemsoseulnt[1o]b)tained by subtractingtheamount of activated sludge added (1.96 mL) from 300 mL] was placed in2 test vessel, then 29.5 a#dLde[dam5o0uan5t1a0dgdievdeoafn3a0nimligne=c2o9n.c5enutrLatxio1n.0o2f21g0/0emm*g/(Ld.ensity)] was collected and ~~) (d ) Slud`gTehebblaasnakl scyuslttuerme (moened:iutmest[ivnesasne l [5]) amount obtained by subtracting the amount of activated sludge added (1.96 mL)from300mL] was placedin a test vessel. (2) Inoc`uTlhaetiaocntiovfatAecdtisvlautdegde Sludge: prepared under the conditions in section 2. above was sionloicduslactoendceinnttroaatbioonvseotfes3t0limqgu/iLds. (b), (c) and (d) in such amounts as to give suspended ~~ Study No. 14101 9 Company Senitized. Does rotcontain TSCA CBY . J (31.)3TCelossteLdiOquxiydgCeunltDuerminagnAdppAanraaltyzuesr:andAmbientConditions `Constant temperature Data processor: tank and measurement unit: ~ Okura Electric Asahi Techneion CTaesrtbovenssdeilo:xi3d0e0ambsLorbienrc:uSbaotdioanlibomtet,leN(om.odI(iffoiercdairnbcoubnatdiioonxibdoettalbes)orption; `made by Wako Pure Chemical Industry) (2) AmbiTeesnttlCioqnudiidtciuolntsu:ring temperature: STteisrtrilnigqumiedtchuoldt:uring period: 2541C 28 days rotary stirring with a magnetic stirrer (3) Conducted at: 511 Kuuro [phonetic] Room 3(1.)4OObbsseerrvvaattiioon:n,Measurement,Etc. during tThheecugletnuerrianlgcpoenrdiiotdi.onOopfetrhateiotenstoflitqhueideqwuaispmviesnutalwlaysexiansmpiencetdedoans apdpariolpyribaasties, ~) (2) Meas`CuhraenmgeenstinofthBeioBcOheDmoifctalheOxtyesgelniqDuiedmwaenrde a(uBtOoDm)at:ically recorded and measured with adataprocessor during the culturing period. The temperature within the tank was measured and recorded on daily basis. ~~ Study No. 14101 10 Company Sanltized. Does not contalnTSCACBF ~ 3.5Establishmentof Conditionsfor Quantitative DeterminationofTestSubstanceby 4 inv`eLstiiqgu`aiCtodeCnddhiurtsoiimonnagstlfoiogqrurtiahdpechhqyur-aonMmtaaisttasotSgiprveaecpdtheryto-emmreamtsirsnyastpieocntorfomtehtertyes(tLsCu-bMsSt)an.ceHowwereever, breecmaauisneintghiisnstthuedtyeswtalsiqtueirdmsiwneatreedonontJcualrryi2e8d,oduettfeorlmlionwaitnigontsheofentdhoeftteshtesucbuslttuarnicneg period. (1) QuIannsttirtuamteinvteaDteitoenr:minationLiCqounidditcihornos:magraph-mass spectrometer HMiagshspseprecftorrommaentceer:liquZidMcDhro(mWaatteorgsr)aph: Alliance 2690 (Waters) `Column: Column temperature: `3L5oClumn ODS (CERI; 16 cm x 2.1mm ILD. Eluant: A: B: 10 mmol/L 10 mmol/L acetic acetic aacciidd//aammmmoonniiuumm//wmaettehransoollustoilountion Flow rate: AB (82 VIV) 0.2 mImin Injection rate: 3uL ~~) MasTsonSipzeacttiroonmmeettehroCdo:nditions `Electrospray (EST) - Detected ions: Detection method: Negative fons Selected ion monitoring (SIM) STeolnescotuerdcieotnesm(pme/rza)t:ure: 514190,5C69, 649, 669, 749, 769 (see Fig. 3) CDeosnoelvvaotlitaogne:system temperature: ~~ 400C 30V (2) PrepAara1t,i0o0n0omfgC/aLlisborlauttiioonnCouftrvhe:tet specimen was prepared by precisely weighing owuitth1m0e0tmhagnoolf tahnedsmpeetchiamneonl/fpuumriisfiheedd,wattheern (d7i/s3soVl/vVin)gtiot ifnormmet1h.a5n0o,l3..0T0hiasndwa6s.0d0ilmugt/eLd. s(1t)anadbaorvde,soaluntdioancsa.liQburaanttiiotnactuirvveedewtaesrmpirneaptairoendsuwseirnegctahrercioendcoeunttruantdieornsthaencdotnhdeittiootanls in `peak areas inthe respective mass fragmentograms (see Fig. 2). ~ Study No. 14101 1 Company Sanlized. Doce rot contain TSCA CBI --~ 3.6 ComtphePeurcetntDiegrandatgion 9 nfeoarrmeuslta.`inTthTeehgpeeer.rrecseunlttsdewgerraedaetxipornesosfetdheasteisnttseguebrsstabnycerowuansdicnogmpthuetfeidrstfrdoecmitmhaefloplllaocweitnogshe. Percent degradation as determined from the biochemical oxygen demand (0D) Degradation (%) = [(BOD - BYTOD"] x 100 where BOD: `Bmieoacshuermeidcvaallouexyignemnildleimgraanmdso(fm(gs)ludge + test substance) system; B: Biochemical oxygen demandofsludge blank system; measured value in milligrams (mg) TOD" tTehsetorseubtsitcaanlcoe;xycgaelcnudleamteadnvdarleuqeuiinremditlloigcroammpsle(tmegl)y oxidize the "2 Culcuined suming aest, roc tyof) nis mm the respective components in the test subinedstfanrceo. mthemolecular formulasandcontenotsf 4. Confirming the Study Conditions Y the BODT,hewedreeregspoerfctaeinvieelliysn5e9d%eagndr76a%d,aftatheetrr7eibdyoacynosnfainrdmi14nd g thaat tahsey detetses trcmoin, ndeitdiofnrsom used in thisstudywere appropriate. A TN: Due to poor legiblyoftheJapaneseeign, subscripts in formula may need 0 be verified, Study No. 14101 2 Company Senitized. Dossnotcontaln TSCA CEL C 5. Study Results 5.1 CondofiTetstLiiqouinds `The conditoifotnhes test liquids are shown below. ElI Asarol -- CT -- cuiarte ---- 52 BMODeasuRresuletsfmorTeestLniqutids BOD and degradation af2t8edayrs areshown below (see Table 1 and Fig. 1). wat+eterst substance)| (sludge + test substance) - [(DBeBgOrDaDd"TMatiT on |Tmm%g|| = 03 [|611s]| 16 [|1576 133=970] TT *4: tVhaelvuaelsuefofrorthtehe(sslluuddggee+bltaesntkssuybsstteamn.ce) systems are values obtained aftersubtracting Study No. 14101 13 Company Sanilized. Ross not contain TSCA nt 2 ~~ Taibtledi1ation table oNr the percentage biodegradation by 300. E|T[oEve |i a [oFos| a[aEsr |pe [Fores| [[a8[ewsa]]ws1 owoslwTr]adaosl] wo|] osre]l_a-]] 77] [T8EToeoo]ass]]emollwwrs] eeaalwsro]] seaall aare]] Deg. : Percentage biodegradetion : Vessel =D no. [I : eg Sludge + Aniline Vessel no, @ @: Sludge + Test substance TVeesstseslubnos.tan8ce of 3+ 0Waxtsevras +saTteesdt. substance }( ~~ Chere of 30D : Pig. 1 | Deg. = [BOD - 3) / [ TOD ) x 100 (%) TOD ofTest substance : $3.9 (mg) 1 FosTonci300%Le1a13 ent 3 589.57 / 610.47 =: 2.13 Ev Yatete 8.75 0a / CaBnN = 278.89 / 83.13 = 3.01 TOD = 80.0 x 3.01 = 80.3 (mg) 2003.07.51 Nome I.Dobe 2 Jly--No. 1--4 1 Gomoanv Senitized. DonotecontsainTsca or. ~ 9 Flea Test No. chart of 1410 sop ( Test substance y ACpupClaptretasatntuiisrnigtico oonnddrioe ns: s No, ONeLZ . etsciyated sludge oi TReefsetresnucbestasnucbestah:c:ei(ioe.niiiliinoeeo)i.s-Activated sludge ":oooiooeoieeos 30 Ged 100(w/2) 100 (ag/2) 30 (mg/2) boistin III B Ra vd a30,2000) *_Beguler contttion Safmkploedesdcsrseirpttptiioon [CO [sides matte |[7t6h6d.a3y|[147t0h2c6e0y0|[(7a2g)412s||i2oTdvheoacaoyy|] [8TeiSluuddcgee =+TTeossttssuubbssttamnmceea]l [0 Tstedee = Test substance] 55:.50|| r6.o6|| 30v.i0s | adogor]] 5.5 | 5.6] 5.7| F057] [ [8 8TTwoaomtterrol+rbeenntisub3)stmme|sl 35.0||oziss ] 40.05|| 05.3]7] Too ~ k3l E ors i {i5 a 3 i = 20 rere reer QED o ERRTT 7 Cutetretc1h4 Tne ( o~ad2t1hier) ' 28 2003.07.31Name 7 Yodisds 2 Study No. 14101 15 Cotmpany Sanitized. Does not contalnTSCA CBY > aTo oe (Solvent blank rept Study No. 14101 one 25a-Jumnr-o2,n0%0s3) `Peak position. Peak position. L" EET os [Standardsolution 1.50 mg/L. L EEE EE TE TM ert -- 25. 2003 has 1.20 [|. araz a| T5a5 ah eEdenn Te Peuk|d Toag ses foaess x 0 857 y-- ES e TE Standardsolution3.00 eh aE ~ EE 25 2003 a rot | oa rakz 1.000 way T5iTm8o STeHont wTTeRnardEea z Th an FA \\.T,e E s i i [Standardsolution6.00 mg p : &1 rat 14101j028. Peak! ot . 800 1400 Too je \ ee 25-Jun-2003 SIRof ChanneESl-s| roe iT4oeooTpEt eSwEdEpmDe ATTHNE Be Tu eRe Ta Rs as (ol lls Tgff S Fig.2-1 Massfragmertograms of LC-MS enalysis forcalibractuirvoen. StudyNo. 14101 16 Company Sanitized. Doesnot contaln TSCACBY 2 oo | Study No. 1410] |. ~ Concentration(mg/L) r=y=30891x lll 300 Toi | [eo Tem] Fig.2.2 Calibration curveoftest item. A October , 2003 Name EVER 4 he . StudyNo. 14101 Company Sanitized. Dos notcontainTSCACBU 17 ~ so tere Instrument MS Waters2D. HPLO :Witars Allarce2880 Servle__Standard soetn $00ma/l. 1010 Contin et om tm Semplesize 2 ul. (Solvent ethene!/purifiedweter (2/3VAD) ore 88. ie Column_LgoOlDSu(OmRDn|Gok elze 18em x20yom1D, Bont A10nmei/ Amorimseststs bomathand 8:[oer Amorke sos amenv byvoter _ 5 coins keinpote ES rn 48 Dorot_toapine_ ProbeESI Copllary _3KV, Desoltvemap.t4i00o'Cn, Dasolgvasat43i0 oL/nh --_ 40d Car_o=niVa, APel prebetam_=,0 ) SewnCom _0_V, BeLVa ,RFr Laos 02 V - `Sours blocktemp._110_C NST lon energy 02V, Mukpl5i5e0 Vr Massrangs(m/e) 400-1000 Sethe_ nm= iz Fig. 3-1 Vir Ti SmeoadFotrd oeno Mess spectra of test item (analytioal conditions). owed ownsTlhelii 5 Study No. 14101 18 (Company Sanitized. Coesnot cc-ontaiinn TSCACEE J [Staianodeard sciution6a0s.s0mg/L / Fakt Stady No. L4101 To-dan 4SanE20s0.3 oeSse Solr % F-- ass o/pakz TM of 1044 ~~ [Stae ndardsolution63a .0mg/L ah a ae wm Tau 2003 1p4101145 480.548)Cm(4750-0384100105) "Soenes | Mampeimalpak1 | i - 1 14101045 78 (5.508) Cm (76:77-{50:70486: 4: Scan ES- Masspofpcec2 4141011045 144 (10.588) Cm (144-{100:140+160:160)) EY 4;Sea2nE2S0. Mat esmofpasks 4a 71 I] x0 108" 1400] 2007.8.13 =. a Fig,3-2 Massspeocftetstri3team, Study No. 14101 19 Company Saniized. Boe net contaln TSCACBY 2 StudyNo. 14101 [1G4l1a0n1d0a4r5d solution 60.0 mall Teusnane, " Tm Totalion chromatogram 3.0808] us re 2Juiz SuPeak | w:o1ts as . Tangssa.l 3 tmnt 0 53% 141019045 L [hr ~ bh = 4SeanES- | 18508] Mam domoef 569 prsa) | Mandrmtnpan f=640 `e826| ei . Eoanes=.. Mas mmmof we =669 fumiae | || tp 2He sI iioioes P SE re Teams + Mssmmmemorairaes2 am 11 . " anes ie |Mass eee oftm 769 ST ah aba 12% kn Tie - 510] 20028, 13 T.thd ~~ Fig. 3-3 Mess specotftresatitem (mass chromatograms). SdsNa. aot 2 Company Sanitized. Does not contain TSCA CBI > Peak1 -- m/z 549 569 ~ Peak2 MW. 570 Fragment ion (M-HF-H)" (M-H)" mz 649 669 MW. 670 Fragment ion (M-HF-H)" (M-H)" Fig. 3-4 Massspectraoftest tem(presumedfregmentsofpeak 1aadpeal 2). = Due: wel. fo. 2 wee: TT. Mudid Study No. 14101 21 Company Senitizsd. Does not conteln 8c cBI d Peak 3 StudyNo. 14101 MW. 770 m/z Fragment ion 749 (M-HF-H)" 769 (M-H)" ~ - Fig, 3-5 Mass speciraof test item (presumed fragmentsof peak 3). wei zeolite 2 vem; Iteil Translation: Language Services F. Metreaud December 3, 2003 StudyNo. 14101 22 Goftipany Senitized. Doss not contaln TSCATEY