Document vBe3ZJLoB5GBpnRDwgy22BVoE

R236- 3317 499 AR 226-3317 . DuPont-13970 ~ TRADE SECRET Study Title H-24616: Hydrolytic Stability of H-24616 as a Functionof pH TEST GUIDELINES: OECD Guideline 111 AUTHOR: Bogdan Szostek, Ph.D. STUDY COMPLETED ON: June 9, 2004 PERFORMING LABORATORY: HE.aLskdeulPloLnabtodreat`oNreymfoourrsHeaanldthCaonmdpEannvyironmental Sciences Elkton Road, P.O. Box 50 ~~ Newark, Delaware 19714-0050 LABORATORY PROJECT ID: DuPont-13970 wok Request Novae {I SERVICE CODE rowel _--_--m---- `Company Sanitized. Doss not conan TSCA CEE 2Hi4yd6r1ol6ytc Stabilityof 24616 as a Function ofpil. DuPont-13970 ~~ GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT L"TahbiosrsattuodryywParsacctiocneduScttaenddairndsc,omwphliicahnacreewicotnhsUi.sSt.entEwPiAthTtSheCOAE(C40DCPFriRncpiaprlte7so92f)GGoooodd Laboratory Practice (as revised in 1997) published in ENV/MC/CHEM(98)17and MAFF Japan Good Laboratory Practice Staridards (59 NohSan Number 3850. "cAThlhaterhatoceutsetgrhsiuztbahsteticoahnnacwreaacwstaedsroinczhaeatriboayncatnweraiIszSenOdo9tb0yp0et0rhcfeeorsrtpmioefndiseoudrndlpaerbrioorGraottooordtyh,LeaaibnnodirttaiahtteoiraoynccoPufrtrahacitcisyocesfttSutdhyae.nddaartdas,isthe considered sufficient for the purposesofthis study. Applicant / Sponsor: `EW.iLldmiunPgtoonnt,dDeeNleamwaoruers19a8n9d8Company USA. --~ `Study Director: SLT oy ~~ SeBnoigodraRnesSezoasrtcehk,ChPehmDi.st of-3Duatne -- 2004 Applicant / Sponsor: DuPont Representative Date --~ EIr E `ompany Sanitized. Does not contain TSCA CBI RHy2d1ro6l:ytc Sability ofH-24616 as 3 Functionofptf -- QUALITY ASSURANCE STATEMENT Dupont13970 Haskell Sample Number(s): 24616 DatesofInspections: Conduct: 29-Jan-2004 Records, Reports: 29-30-Mar-2004; 2-Apr-2004 Dates Findings Reported to: Study Director: 29-Jan-2004; 7-Apr-2004 Management: 29-Jan-2004; 7-Apr-2004 --_ Reported by: re `Matthew Eoefil Quality Assurance Auditor 27-JuDaiea-zof Company Santized. Doss not contain TSCA CBI BHyedsr:ol Subily of24616 as a Function ofpi. --~ CERTIFICATION DuPont13970 We, the undersigned, declare that this report provides an accurate evaluationofdata obtained from this study. Approvedby: Eg 5NikKMe ienayger, -- ntsmcs otheFophSn SouCt, PLD. SenoResearchChemist 08=ToDuaed=Zoo o1-Dgauew~20 - -- =A -- Company Sanilized. Does not contain TSCA CEi acteletic Sublyof 2461614 Fusionoft. 7 `TABLE OF CONTENTS Denis Page `GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT cccvssesemsmsmsnininnrensd ETT PRRBORIEL mmrmimmmmssmismisssmisrpmsmermsmomnssssssrse) - 5xFE foPrepare ns------I--I--I pTI --r"ry PT -- ---- 5L Rrasii orgra l II IISI ----I -- S Cl so FT ------e ---- RESULTS ANDDISCUSSION cvssssssssnssssosonnnn18 RECORDS AND SAMPLE STORAGE w.cccocuvmusmsmsmmmmmmmsmmmmmssssssssssssssssssssssssssssssmsssssssssssssossi|d ~ espneet pe ------ Company Saniizad. Doss nt contain TSCA CBE HHy6d1ra6te Sabiliy of24616 a Functionofpif --_ LI OFTSABLTES Dupont13970 Page 1. o Analytical rese ults or aqueous se abilsyeof2e 4616, pH 1.2, 37C, Days 0 12-Feb-2004) nd S 23 2. Analytical r0esf5our(sl1quFteeobsns30s0a8b)ilityofr 24616pi,4.r 0pias buter , SUC, Day0s 2 3. Analyticalerensul2t0s0f4o)aqueous stabilie ty of 24616, pHr 4.0 iratebuffer,r SOC, Days 0 (5-F: eb-2004) 33 4. (An0a3l-yFteibc-a2r0e0s8ults frorraqrueous sail ofH-24616, pH 7.0, SC, DAayAs A0 29-Jan2004) anTdS -- 5. (AOnFalaytbic2a00r8e)s.ultr s or aiqueougs sabiltyofr H24616r , pH 9.0, SO, Day0s (29-rJoan-200w a4n)d S -- 6. Measured concentrationos 7. Measured concanratioonsf or pH 1.2, 37C,Days 0(12-Feb-2008) nd 5 (17-Fe5-2004) 28 PH.40, C, pH 4.0 phatevbruffer, SOm, Daayt0s on M(e5-s7a300c4)o0n5 c(10n-Feb-l2004r) 140,57 140 i S76 D0 --_ 5. PAREN rr Meanred conenuion RRB p70 SC,Day 025-10 2008) 0d (BF 10, Messrsconcenrions GEREN:190, 5, Des 0 09-1an2000) 1 04). 51 035209. 11. Dailytemperaturereadings oftheSHAKINGWATBah. 12 PH ICSU forDAY 0 B00 DRY S.rrros 33 34 --_ J = ---------"-"--m Company Sanitized. Does not contain TSCA CBI Hyd4r6o1l6y:tc Stabilityof24616 a a Function sof pH Duron 13570 -- LOIF FS IGUT RES Page 1. TEuxoarmipnlaete*dFacNtiMveRinsgpreedcisenotf1re0c5o0vemregd/iLnTflHuForienxattreadcatc(tBi),vainndgraecdeiteonntitcrailliebreaxttiroanctsi(aCn)dfaorrdD(Aa)y, , PH 40 phthalatebuffer... a ee asm-- 2. PEMHoxoarm1ip2nlbteuef"dfsFecrNi.vM.e.Risnpgreecdiientofr2e1coe0vmegr/eeLd.iunrTiHnateextdraacct(i3v)e,inngdAresditenocnaleraesxotrnacstta(nCd)arfdor(DAn )a,y 5, S] 3. Eacxtaimvepliengr"eFdNieMntRrescopveecresdoifnTa HblEanxkiccoanttro(l3)saampmlefpofrolDraDeyay5,0p, ipH1212buBfUfeTr (4() nBd thce flcurinaied 38 4. DEHxa4mp1leCr"aFteNbuMfRfesrp(eAct)rsanodfDthaeyf,luopriin4a.t0edCaHcltieveBiUnTgrEed(iBe)n.t.recoveredeicn THE extract for Day 0,en) 5. pExHa7m.p0lbeuf"fFerN(MA)RasnpdeDcaty5o,fpthie7l.u0obrilnaetd activeB).ingredientpreacovered in TH exace t for DaA y 0, ) 6. PExPa8m.p0le "TFe N(MA)R30s0peDct8rao5f,tPhH 9f.l0uBoriUnatTedEsive(iBngrrredrient recovered in THE extract forDay 0, a 7. ReprecalsibreationncutrvaefotrfiRRvY..e..... sns---- stg 5. inRtepermeaslesnttaatnidvaerdc.hromatogramsof 102 aQEtiEot n standaamrd nd 520 1RY i) ~~ 5. 0 Represen8 ative 5 chromatog3 rams r0 sfo]rDa, y5,pH 4 caStaenb.uffer hydrolytic samilse sample e ts 10. Representative chromatograms or Day5, pH 4.0 ciate ber blak sample and 520 yo {SNINRY. --_ Company Saniized. Doss not contain TSCACBU pLastes osm re STUDY INFORMATION Synonyms/Codes: + H-24616 n Composin} Haskell Number: 24616 -- Known Impurities in Total SolidsT : E. scl Chasis: Opecta iqsid Sponsor: WLE.Lii)dunPoont dSe Nwemoeurs98an9d5Company Suds nieCompltsls Decsbr 3, 003 se repr over pags) Experimental StartCompletion; December 22, 2003/ March 2, 2004 = Cory et. boss notTGAaCal HHy2d4r6ol1y6tc Stability ofH-2461a6s a Functionof pH ~~ STUDY PERSONNEL Study Director: Bogdan Szostek, Ph.D. Analytical Associate: Keith B. Prickett, B.S. Management: S. Mark Kennedy, Ph.D. Report Preparation: Wanda F. Dinbokowitz Management: Kim D. Birkmeyer, M.S. DuPont 13970 --~ ~~ -- any Sanitized. Does notcontainTSCA CBI Hydrolytic Subility of1.24616as a Function ofi Dupo 13970 ~~ SUMMARY Hydrolytic stability of H-24616 was tested in buffered aqueous solutions at pH 4.0, 7.0, 9.0 at 50C and pH 1.2 at 37C for five days. Concentrationsoffluorinated active ingredient of 'DHa-y24061a6ndweDrae $ymearsepulriecadtebysa"mpFlNesMfRorsppHect1.r2o,sc4.o0pyphatnhdalpaetrec,e4n.t0rceictorvateer,ie7.s0,we9r.e0 bcuaflfceurls.ateTdhfeor average percent recoveries for the Day0 and Day 5, pH 1.2 replicate samples were 104 3.6% and 86 = 4.0%, respectively. The average percent recoveries for the Da0y and Day 5, pH 4.0 phthalate buffer replicate samples were 93 = 2.9% and 82 + 6.9%, respectively. The average percent recoveries for the Day 0 and Day5, pH 4.0 citrate buffer replicate samples were 92.4 3.2% and 89 3.1%, respectively. The average percent recoveries for the Day 0 and Day 5, DH 7.0 replicate samples were 97: 2.0% and 91 + 1.8%, respectively. The average percent recoveries for the Day 0and Day 5, pH 9.0 replicate samples were 98+ 4.2% and 92 = 0.81%, respectively. The fluorinated active ingredient ofH-24616 was demonstrated to be hydrolytically stable at pH 9.0, pH 7.0, and pH 4.0 citrate buffer witht >one year using the criterion ofDay 0 and Day 5 fluorinated active ingredient recoveries differing by no more than 10 percent. The data obtained for hydrolytic stability offluorinated active ingredientofH-24616 at pH 1.2 and pH 4.0-phthalate buffers is not conclusive. The difference of recoveries for fluorinated active ingredient between Day 0 and Day 5 samples was larger than 10 percent. The data indicates that a significant portionoffluorinated active ingredient stays dissolved after additionof acetonitrile to the samples that were exposedtotest conditions in pH 1.2 and pH 4.0phthalate buffer. Accurate quantitationofthe dissolved fraction of fluorinated active ingredient ~~ `was not achievable for acetonitrile extract. Therefore, accurate mass balance of fluorinated atchteisveepiHnsgrdeiddiennottcionudlidcantoetabneyacchhainegveesd.ofTthhee 'f*lFuoNriMnaRtesdpaeccttirvaeofitnhgreedaiceenttonmiattreirlieale.xtracts for `The measured concentration of id not indicate any hydrolytic degradation of the fluorinated active ingredient material t for pH 4.0 and pH 7.0 buffers. The observed concentrations o---: `be explaimed by the residual resent in the test substance (H-24616). Fof pH 1.2 and 9.0 an increaseofthel oncentration beyond that accounted for byreo se in the test substance was observed. It was estimated that this increase would represent hydrolysisof 0.06 and 0.03% the test substance for pH 1.2 and pH 9.0 buffer, respectively. --~ --- SDLDESE.-- ESP--P-ENI D .:Fbr Company Sanitized. Doesnot contain TSCA CBI 1Hy2d4r6o1ly6tc Stability of24616 a Function ofp DuPont 13970 INTRODUCTION `The purpose of the study was to investigate the hydrolytic stability of H-24616 in pH 1.2, 4.0, 7.0, and 9.0 aqueous buffer solutions. The study was conducted according to the protocol: "Hydrolytic stability of H-24616 as a functionof pH", which was patterned after the OEC] Guideline 111." Additionally, the gencentration ofthe as monitored to investigate if s released from the test substance during the test. STUDY DESIGN A preliminary test was performed on the test substance at 50% 0.1Cat pH 4.0, 7.0, 9.0 and at 370.1C for pH 1.2. Ifless than 10% test substance hydrolysis was observed afe5r days, the test substance was considered hydrolytically stable and no additional testing was performed. In `addition to the test substance determination during the preliminary test, the samples were analyzedfor[J `The study design complies with the following test guidelines: OECD Guideline 111%"with the exception that the OECD Guideline 111 requires that the amountofthe test substance used is the smaller of 0.01 M orhalfthe saturation concentration. The test substance fluorinated active --~ ingredient [AMBRE is csscnially not soluble in water. Therefore, the study was conducted With excessofundissolved fluorinated active ingredient and the amountofthe test substance was sufficient to satisfy the sensitivity requirements ofanalytical method. The molar concentration of fluorinated active ingredient was estimated to be 0.002 M (in 5. Calculations). MATERIALS AND METHODS A. Materials i Test Substance Name: 21616 =[ \ Haskell number: 24616 The test substance was characterized to determine % fluorine, % solids, pH, Mn, Mw, and `polydispersity with the following results: -_ otipany Barliized. Docs not containTSCATE 2Hy4d6r1o6ly:ie SiabilityofH-24616 as a Function.ofpil om\ %%FShoolridisne(w(/wu)i) 2033%% oi 63 MiMaw: 11750000 aammuu polydispersity 113 2 ResiduaYln 24616 DuPon13970 rr in the test substance (H-24616) was determined by LC/MS using the OsfamHe-2a4n6al1y6ti(caaplpmreotxhimoadteaslyde4s5cmrgib)ewdeirneAnianltyrsoidsucoef{d tlo gliassnvialMs,etwheoidgsh.ed,Fiv1e0 rmeLpliocfatTeHsFamples (tetrabydrofurane) was added, vials crimped, and vortexed for 10 minutes on a mulfitube vortexer. A 0.1 mL aliquotofdissolved sample was transferred to a glass LC vial and 0.9 mL acetonitrile and intemal ee added. Calibration standards were made in 10/90% THF acetonitrile in the range from100 to 1020 ng/mL[RSNNEY 3. Prepoafrbufafetrsoiluotionns The following buffer solutions were prepared as described in the Annex to OECD guideline 111.0) a pHIZ ~ `STihgem0a.)2iMn 2p5o0tamsLsioufmwachtleorr.idTeh(eK0C.I2) Mwashypdrreopcahrleodribcyadciisds(olHvCiIn)gw3a.7s8pgreopfarKeCdIb(ypudriiltyut9i9n.g0%4.,93 g opfrehpyadrreodchblyorpilcacaicnigd 1(2356.m5L-3o8f%,0.S2igMnaK)Ciln a2n5d0m16L1.w3amteLr.ofT2he0.p2HM1.H2CbIufsfoelrutsioolnutiinotno waa5s00-mL fhlyadsrkocahnldorbircinagciindg the flask to volume with water. The pH of the buffer was adjusted to 1.2 with bo pH4 P1h0t.h2a1lgaotefbpuoftfaers:si0.u1mMhydsorlougteinonpohfthpaoltaatses(ipuumrihtyyd9r9o+g3e%,n pShitghmaal)atienwSa0s0pmreLpoafrweadtbeyr.disTshoelvpiHng4.0 obuffafe0r.1soMlutpiootnaswsaisumprheypdarreodgebnypphltahcailnagte2smolLutoiofan i0n.t1o M2 5s0o0d-imuLm fhlyadsrkoaxniddberisonlguitnigotnhaendfl2a5s0k tmoL volume with water. The pHofthe buffer was adjusted to 4.0 with hydrochloric acid. oCfitproattaesbsuiffuemr:di0h.1ydMrogseolnuctiioantoefp(poutraistysi9u9m+d%i,hFyldurkoag)eninci5t0r0atemwLaosfpwraetpearr.edTbhye dissolving 11.5 pH 4 buffer g s0.o1luMtiopnowtaassspiruempdairheyddbryogpelnacciintrga4te5smolLutoiofnai0n.t1o Ma 5s0o0d-imuLm fhlyadsrkoaxniddebrsionlgutiinogntahnedfl2a5sk0tmoL ofa volume with water. The pHofthe buffer was adjusted to 4.0 with 1 M sodium hydroxide. --_ --_-- `Company Sanitized. Doss not contain Tsca car HHy2d4r6o1l6y:te abilityofH-24616 as a Function ofpil. --_ c pHT: DuPont13970 `pThhoesp0h.a1tMe, smoolnuotbioansoifcp(oKtHas;sPiOusm, ppuhroistpyh9at9e.5w%a,sSpirgempaa)reidn 5by00dimsLsoolfviwnagte6r..81 TghoefppHota7bsusfifuemr solution was prepared by placing 148 mL ofa 0.1 M sodium hydroxide solution and 250 mL of t`whieth0.w1atMer.poTtahsespiuHmopfhotshephbautfefesrolwuatsioandjiuntsoteadSt0o07.-0 wmiLtflhashkyadrnodcbhrlionrgiicnagcitdh.e flask to volume a pHO: `The 0.1 M dissolving boric acid (HBOs) solution in 0.1 M 3.73 g of KCI (purity 99.0%, Sigma) potassium chloride (KCI) was prepared by and 3.09 gof HyBO; (purity 99.5%, Sigma) in 5so0l0utmioLnoifn w0a.1teMr. KTChleapnHd 91.006.b5ufmfeLr sooflut2.i0o.1nMwassopdrieupmarheyddrboyxipldaecsionlgu2ti5o0nminLtooafa5000.1-mML HflsaBskOs `and bringing the flask to volume with water. sodium hydroxide. The pHofthe buffer was adjusted to 9.0 with `puWraitfeircuastieodnfsoyrsbtuefmf.erTphreepawraatteironrewsaisstiovbittayiinse>d1f8r.o2mmaegBaaonhsmteeeamd.NAANlOlpbuufrfeerDsiolaumtioonnds wwearteer sterilized prior to use by filtering through Corning Sterilization Fier systems with 0.22-um cellulose acetate filter. The buffer solutions were stored at room temperature. 4. Equipment --~ pWaHteMertBera:t POrreicoinsiMoondReelci2p5r0ocAal Shaking Water Bath, Model 50, Precision AAuuttoocmlaaiviec:Pips: AReMseSaCroOh MPorode1l003,0210300, 5000; Eppendorfl CeBnatlrainfcueg:e: MSeotrtvlaelrLAeEg-e1n0d0RT VoGrlatseserVi:als: M10ulmiLtubboreosviolriecxaeter,SeVrWumRType Reaction vials (Supelco) with 20 mm Barrier Septa 3(0FomliSnilbeifcoornee)u0s,.e1. thik (Supelco). Th vialsandseptawere autoclaved st 121C for B. Methods 1. Preliminary test 3T7heCprfeolripmHina1.r2y. teTsht ewassampeersfaomrpmleed pornepHa-ra2t4i6o1n6partoc5e0duCreataenadcshtoufdtyhdeespiHgn4.w0a,s7.f0o,l9l.o0weadndfoart cauatcohoclfatvhede gplHassstveisatlesd.anAdpp3rmoxLiomfaatpeplryo4p5rimatgeofbutfhfeerHs-o2l4ut6i1o6n wwaass waeddiegdh.edTihnetovisailxsteweenre10c-rmiLmp cwaepreedpwriotchestsheedaflourmainanluymsifsoialsctohveerDeadyp0arstoamfpltehse.seRpetmaafianciinnggienisgihdtestahmepvliaels. wEeirgehtwroaftphpeedviwailtsh dthaeysalatumthienuapmprfooiplr,isautebmteermpgeerdatiunrteh.e wEaitghetr bviaatlhs,caonndtasihnaiknegnoantly1030mrLpomfianptphreopwraitaetrebbautfhffeorrwfeirvee --_ prepared to serve as the blanks. Fourof them were used as the Day 0 blanks and the other four -_-- Company Sanitized. Does not contain TSCACBI HHy2d4r6o1l6y:ic SabilitofH-24616 asa Function ofpi Dupont 13970 --~ vwiearles cwornatpapiegidngin3Aml LfoiolfaanpdprpolparcieadteinbutfhfeewrawteerrebastphitkoedbB e the r Day b) lanks. sAodlduittiioonna(l4l.5y,Lsixof p 1020 s "The vials were crimp capped with the aluminum foil coveredpart ofthe septa facing inside analysis on Day 0. tThheevioatl.herThtrhereeeofvitahlse vials were wwrearpepuesdedinaasltuhmeiQnCumsfaomipllaensdoplarceld i)n th.e wa t er bath together with cightofthe test substance vials. These served a roleofQC samples for C-- were processed on Day 5. Fourofthe for the fluorinated active Day ingredient 0 or fooufrofFt-h2e46D16ayde5tevrimailnsaatinodn.twTohbelaontkhevriaflosur of the Day 0 or Day 5 vials, two blanks vials, or-- vials were processed for the [_Sm--_y:cccrmination. 2. pH measuremeantd temperature dTehseigpnHatweadsfomretahseuHre-d24o6n1D6aaynal$yisnist.heIvniaadlvserctoennttaliyn,ionngltyhepbHoufffetrhaenbduftfheerstewstassumbsetaasnucree,d on Dtheeyp0H, neoltectthreodpeHwoafssaimntprloedsucceodnttoaitnhienvgiatlhse atensdt tshuebsptaHncree.adiDnagyw5assatmapklene.viTahlsesweerviealusncwaeprpeed, `fTuhrethweartperrobcaetshsehdasfoarnHa-u2t4o6m1a6tiacnatleymspise.ratTuhree tceomntpreorlatbuuirletofitn.heThweatreeradbiantghswoafsthmeonteimtpoerreadtudraei:ly. control were checked and recorded daily. 3. Analysisoftest substance f~ VwiearlesrdeecsaipgpneadteadnfdorvoHr-t2e4x6ed16foarna1l0ysmiisnuwteerse. uAndcdaiptpieodn,o6famceLtoonfiatcreitloenittortihleevwiaalscaodndteadi,nivnigaltsest sTuhbesvtiaanlcsewseursepecnednetrdifinugaedbuffofrerfisfotleeuntimoinncuatuessesatt3he20f0luroprmin,atuendcaapcptievde,ianugdretdhieenltiqtuoipdrpechiapsietawtea.s rwaesmocvaepdpefdr,oamntdhethveianl.sonTiecnatmeidllfiolirt3er0somfintuetters.ahyAdpropfruorxainmeat(eTlHyF)1 mwaLsofatddheedTtoHFthesovliualt,iotnhewavisal panlaalcyesdisi.naThgelasbslaLnCk sviaamlp,l0e.s1 wmeLreofsmuebtjheyclte-ddst-oatlhceohsoalmewapsroacdedduerdea,nwditshuobujtecpteHdmteoa'suFr-eNmMeRnt.. TTehne msitoncuktessooluftsioonnoicfatHi-o2n4w6a1s6 rweaqsuimraeddetobcyodmipslseotlevliyngdi0s.s4o5lvgeotfheH-te2s4t6s1ub6stiannc1e0.mLCaloifbTrHatFi.on csatlainbdraartdisonfosrtatnhdeaHrd-s24r6a1ng6eadnfalryosmis20w0erteo m2a0d00e mbyg/aLpporfopfrliuaotreindaitleudtiaocntiovfetihnegrsetodcike.ntoTfheH-24616. FLirneesahrsrteogcrkessosliuotnoifontahnedpceaalikbraarteiaovns.stsatnadnadradrsdwceornecepnrterpaatrieodn wfoarseuascehdintosteasntcaebolfisahnatlhyesicsa.libration curves. ~~ J Company Sanilized. Doss not contain TSCA CBE HyH2dy6rdr1oo6lly:tcSSutbabiilliityy ooffHH-4:2G4I6616asaas FauFnunccttiioonnoofflpt ~ Instrumentation and conditions DeDupmonti139n70 NMR instrument: Acquisiion Parameters Procesing Parameter: `Bruker AMX360 with a F/'H dual probe PTuilmsee DPoromgarianm:Si2z1e: 16k DNI uDelmayo:bfS1eca0nrss: 128 PDuulmsmLyenSgctanh: 3psec IRnescteriuvmeernGtiFnr:equaeuntcoym:at3e6d.b8y3t14h5i6n6siMruHmzent WSiizneodofRweFaulncStipone:cErMu 16k Window Size S0Hz Peak Used for Quantaton CF, resonancat 32.5 ppm 4 Analysis orEEE Samples designated offs were processed by injecting 3 mL methyl-tert-butyl ether (TBE) through the vial septum using a glass syringe. `were uncapped before MTBE injection to avoid any losses of Neither Day 0 nor Day samples The vials containing ~~ winejreectuendcMapTpBedEawnedre0.v1omrtLeoxefdthfoerM1T0 BmiEnuetxetsraocnt waamsulttriatnusbfeerVroerdtetxoear.glaSsusbsLeCquveinatl,ly0,.9thmeLvials `acetonitrile added, spiked with internal standard, crimped capped, and subjected to LC/MS analysis forJD.) The] used as the analytical standard of Oakwood Products, West Columbia, SC) was The 10 mg/L)of the analytical st [was usedasthe internal standard. and the intemal standard were Stock solutions prepared in methanol and were stored refrigerated. The calibration standards were prepared fresh for each calibration in 11000/0900p%igM/TLBEY/acNeto]nitriAlec.onsTthaentcalleiveblroatfiionntsetrannadlasrtdasnwdaerrde wmaasdeusiend:the52r0anvgevf-- rom 100 to The calibratio`npecaukrvaeresawfeorreicoonnstmroanctietdoruesdinfogthe ratio of he ipdeathkeacroenacefnotrriaotinosns. omz fmonitored for ee ae e --t `Company Sanitized. Does not contain TSCA CBI HHyHHydudrsroo12ll6y:tc4 Saabbilii6tyooffHl-24o 616 aua Fusncioa nofitfuncionolppDuuop_ wonits13_ o9n7 ~ Instrumeanndtcoandtitiioonsn: HMPSLICnstnrsutmuemnte:nt: 2MQo,deMlic2r79o5m,asWsates CoLCluPamra:meters: Mobile hase: LAc vua3tueCrn18a2),20mxm 2mm GBr-a"dmTieietmnheat(nmoiln) wa un 01 3300 07 25 00 110000 Column Temperature: 30C 6s1 3020 3n CInojleuctminonSVowluime: 22a0n4d1 min MTo5niPzaatriaomnetmeordse:: Elecospray, negative ons CCaopniellvaoruygveo:lage: 2to5v% A DDSueosumorlcAveactTqieionmiTopenemprFearaa:tnuoren:: _L310200CAI of, 5R 2,05 min CO 5.01467, 325 mz. 06 min 5. Calculations Flow 0(2n5min) 002255 002253 025 Tflhueorfionlaltoewdiancgticvaelciunlgarteidoinesntwe(rTeabulseesd1-t5o).detFeorrmeixnaemtphlee,niomfi0n.a0l45cognocfenHtr-a2t4i6o1ns6 ocfonHt-a2i4ni6n1g622.6% Mthueornionmaitneadlacctoinvceeniinrgarteidoineonft wfalusorwieniagthededacititvoetihnegrveiadlieanntdi3: m(L0.o0f45bguf*fe0r.2w2a6s*a10d0d0edmgto/gt)he/vial 0th.e00m3eaLsu=r3e3d9c0onmcge/nLtroaftfilounobryintahteednoamcitnivaelicnognrcedeinetnrta.tioTnh,ei%mersec1o0v0e%r.y is calculated by dividing `fTohleloewxsp:etchteed4.(5nopmLinoafl)10c2o0ncmengt}ragion of pike reipnrtehseenstpsi4k.e5d psaLmxpl1e0s2w0ansgc/aulLc=ul4a5te9d0ansg hon by dividing thetmhaeemaoMsuTunrBteEdwacesoxntercxaetcnrttarciast:tei4do5nw9ib0tyhntg3h/em3 nLmoLmo=if1nMa5Tl3B0cEonn.cge/nmTtLhr.eateiTxohpnee,ct%tiemdreenscoo1mv0ie0nr%ay.liscocnaclecnutlrataetdion m`oThleecmuollaarrwceoingchetnt(rMatii)onoffolfufolruionraintaetdedacatcitvieveinignrgerdeideinetn:t1c5a0n0bge/meosltei.matWehdeunsi4n5g mavgeroafgeH-24616 iisngurseeddie,nit. reTpareksienngtsin(t0o.a04c5cogun*t0t.ha2t236m1L5o0fgbiumfofleer =is 6u.s7e8d 1t0h%is mwoolueldofrefplrueosreinnatttehdeamctoilvaer --~ concentration: 6.78 10 molel0.003 L = 0.0023 M. The expected concentration ofS a. in Company Saniized.Dossnotcontain sca cay Hdl sbi of 1.24616 3s a untofoit. Dupont1397 ~ tfhoellMowTiBngEwaexyt.racTteinf p1e0rc%eonftothfe4H5-m2g46o1f6Hh-y2d4r6o1l6yzreedprteosfeonrts the test substance contains 4.5 mg. Takinbgenetsotiamcateeqdunintthee FthiermhydroW lisisao ife 4.5 mmgg1/2omf.tm3ho%elfetl)euto=r0sinu.eb7sa9tn2adnacsssmueomueiln'dg)ltehaa`tTshai:ls contained in 3 mLs ofMTBE extract. Therefore, expecic 4fl.u5omrinxe is0.p1r2e3se(nt35as4 amount of -- To t ould be: 0.79 mg/0.003 L= 263 mL. concentration of ould be: 4d`e5ThremivgHe-ds2af4mr6po1lm6eroceofsnidtHua-ai2ln4s6r1e6siidsu:a4l5mgtheofMHT2B4TEh6e1ex6etxrxpaec(ct0t(e.3d0,1m9Lm%)a/,x1ai0s0ms)uu0mm.i0cn0og8nccLeo=nmtp3rla6et5tieomneaoxtft.rEactiEon Rfor a (2850 pg/L). --_ --~ i" `Company Sanitized. Does not contain TSCA cal HHy2d4r6o1l6y:tc StabilityofH-24616a a Function ofpi . --~ RESULTS AND DISCUSSION Dupor-13970 Tables 1-5 present the results for measured concentrations of fluorinated active ingredient of H7.-02,496.01,6raesnpdecpteirvceelnyt. rTechoevearvieersaogbetpaeirnceedntforreDcoavyer0ieasndfoDratyheSDraeypl0icaantedsDaamypl5e,spfHor1p.2Hre1p.2l,ic4a.t0e, samples are 1042 3.6% and 86 = 4.0%, respectively. The average percent recoveries for the Dresape0yctiavnedlyD.ayTh5e, pavHer4a.g0ephptehracleantterbeucfofveerrireepslfiocrattehesaDmapyle0saanrde 9D3ay=52,.p9H% a4.n0dc8i2tr+ate6.b9u%ff,er replicate samples are 92 = 3.2% and 89 + 3.1%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 7.0 replicate samples are 97 2.0% and 91 + 1.8%, respectively. The average percent recoveries for the Day 0 and Day 5, pH 9.0 replicate samples are 98+ 4.2% and 92% 0.81%, respectively. The differenceofaverage recoveries between the Day and Day 0 samplesofthe fluorinated active ingredient was less than 10% for pHs 4.0 citrate buffer, pH 7.0, and pH 9.0. This indicates that the fluorinated active ingredient ofH-24616 is hydrolytically stable at these pHs (tz >one year). `The hydrolytic stability experiment for pH 4.0 was done with two different buffers: phthalate and citrate (Table 2andTable 3). Two different buffers were used as initial experiments with phthalate buffer indicated that the buffer composition affects the solubilityoffluorinated active ingredient in a buffer when subjected to prolonged exposure (5 days) at elevated temperature (50C). Simply, the additionofacetonitrile to the vial containing the test substance would not -- completely precipitate the fluorinated active ingredient. The amountofthe active ingredient recovered in the THF extract would be 30-50% (Table 2) for the phthalate buffer. Inorderto determine if the unaccounted portionofthe fluorinated active ingredient was present in the acetonitrile extract, the NMR spectrumofthe extracts was collected and a quantitationof the fluorinated active ingredient in the acetonitile extract was attempted. Accurate quantitation of the fluorinated active ingredient in the acetonitrile extract (acetonitrile extract contains 6 mL acetonitrile and 3 mL ofa buffer) was not possible because the calibration standards of fluorinated active ingredient in the acetonitrile extract could not be prepared using the THF stock of fluorinated active ingredient. The fluorinated active ingredient would precipitate ifintroduced 10 the acetonitrile extract using THF stock. In order to determine the sensitivity difference. between the NVIR ments done for THF and acetonitrile extract matrix, a calibration standard og vo solvents was made and measured by the NMR methods. This allowed the derivationofthe sensitivity-difference factor that was applied to correct for the `measurement of fluorinated active ingredient in the acetonitrile extract vs. the calibration standards made in THF. However, this procedure is considered to give only semi-quantitative results. Table 2 results indicate that significant portionofthe fluorinated active ingredient is pDraeyse5ntTiHnFtheextarcaectto,niDtariyle$eaxctertaocntitfroirleDaexytr5acsta,mapnldesa.caCliobmrpaatriiosnosntoafntdahred 'i'nFdiNcaMteRs tshpaetcttrhae of fluorinated active ingredient present in the acetonitrile extract did not undergo a chemical change that would be visible by F NMR (Figure I). The average recoveryofthe fluorinated active: ingredient for Day S samples is lower by 11% (Table 2) than the recovery for Day 0. However, taking into account the semi-quantiative natureofresults for acetonitrile extracts, it is difficult --_ to draw a conclusion about hydrolytic stability (t2 >one year)ofthe fluorinated active ingredient as the difference may be coming from the inaccuracy of the results obtained for the "18 Company Sanitizsd, Dassnotcontain TSCACBI SHHoy2d4rt6o1let6icSStaebilyityoorf Ht-i2z4616 u15saaFtuuncdtiioononfoplipt i DDuupoomnnti13o9m70 acetonitrile extracts. The observed solubility issues can be linkedto buffer composition, not the ~~ bbuuffffeerr'(sTpabHlefo3r).the pH 4.0 buffers. The solubility issue was not observed for citrate, pH 4.0 tThhaend1i0ffpeerrecnecnteo.fHaovweervaegre,resciomvielrairelsy tboetpwHee4.n0t-hpehtDhaalyat5eabnudffDera,yth0essaomlpublielsitfyorispsHues1.w2eirsemore aecntciovuenitnegrreeddifeonrttwheaspHdet1e.2ctbeudffienr.theTaabcleteon1itirnidliecaetxetsratchtast,abusti,gnfiofrictahnetraemasoounnstdoefscfrliuboerdinaabtoedve for texhterpahctthwaalastenobtufpfoesrs,ibalcec.urTahteerqeufaonrtei,tatthiisondoafttahweasflnuootricnoantcelduascitvieveabionugtretdhieenhtydirnotlhyetiaccesttoanbiiflriitlyeof `thTeHFfleuxotrriancatt,eDdaayct5ivaeceitnognrietdriielneteaxttrpaHct1f.o2r.pHHow1e.v2ebru,ffecro,mpaanraidsocnaolifbtrahteioNn MstRandsapredctirnaodficDataeyd 5 that the fluorinated active ingredient present in the acetonitrile extract did not undergo a scpheecmtircaaol fcahabnlganekthsaatmwploeuladndbethveisDiablye 0byTHFF NexMtrRact(FfiogrupreH2)1..2.FiFgiugruere3sp4r-e6sepnrtessetnhte NthMe R comparison PH 7.0, and pofHN9M.0Rbusfpfeecrts,rareosbpteacitnievedlyf.or Da0y 2nd Day 5 THF extracts for pH 4.0 citrate, The content ors study. Based on the tl resultsoffive H-24616 was measured before the startofthe replicatesofthe H-24616, the content Cr hygroltic, "H-2W 4616R was ctahlecutleasttesdutbostbaenc0e.0d1u9r:i0ng.0t0h0e3t%i.meoAfsshuymdironlgyttihacttsthaebbiluiftfyeerxpeexrtiramcetnst,ailtheefxe preescitdueadl hcyodnrcoelnytsriastoifotnhoefY fluorn inated the test system would be 2.85 active ingredientofH-24616 mg/L (see 5 Calculations). wouldgnost likely result in The the release. ~~ coTonafQcbelnIterN6at1i)0onprIofeftsBeennp--tehrec_empetaosfutrhHee-d2te4cso6tn1sc6yesnhtsyeadtmrioowlnoysuozledRdbreel2Ue6as3inmgBg/L oc D(seye 50Ctalcublaterixopnse)ic.tecd t os-- amplo es for r pH P1.H2,14..20(pThatbhlaela6t)e,in4d.i0cactitersatteh,e7i.n0c,r9e.a0s,eroefspectively. The measured concentration oncentzation for Day 5 samples beyond substance. The the level that concentration can be accounted for by the resid casured for the Day0 samples is resent in the test comparable with the expected congentration of riginating from eons Ge Day 5 concentration `concentration oor-- iginating fro`mhitghheerrebsyidaueppraximately 1T.r5emogbs/eLrvemdoreintchraenatsheeoerxpected creopnrcoesnetnrtastioonnlyinedsitciamteastetdhehyhdyrdorloylsyissiosoffaptphreotxeismtastueblsyta0n.c0e6t%o o(1.v5pmg/L--/ 26--30`Tmhigs/iLnc*re1a0s0e%) of the test substance present in the test system. The acceptable recovery resus control samples (QC) for Da0y and Day 5 demonstrate the validity forthe quality nr (Table 6). The measured concentrations o-- PH 4.0 citrate, pIP4,0 phthalate, pH 7.0 test the e systemgp nod t exceed signi7-f9i)c.antTlhyisthiendeixcpaetcetsetdhactontcheenretriastnioonocorntirigbution o inating from gon the hydrolysis ofthe test acceptable range (Tables substance. 7-9). The The recovery results forQC samples are also measured concentration fe PH 9.0 the (Table 10) aicndciocuantteetdhefoirnbcryeatshee oresid onceptresteinotnifnorthDeatyestS ssuabmsptlaencseb.eyTohneditnhcerelaesveelisthat can be -- approximately 0 Qui iis .85 indi mg] cative of eyondghe congentration generation from hyd explainable by the residual rolysisoffluorinated activ e a Company Sanitized. Does not contaainin TSCAACTB: eH2yt 4d6r1ol6y:te e SabiltyooffH-R2G46i16csa etFuundcitinoan lofp. DDuuPoonwtz.1o39n70 ~ ingredient at (0:85 mg/L / pH 9.0. This 2630 mg/L * increase represents 100 %)ofthe test s an ubs estimated tance pres hydrolysisof ent in the test approximately system. 0. 03 % Fcihgruormeat7osghroawmss aforreptrhegentativegalibbrraattiioonncsutravnedaorbdt,aDinaeyd5o, pHl4l.0 t)est sFaimgpulree,san-d10Dashyo0w, PH 4.0 blank sample, respectively. Table cases, t1h1eswuamtmearrbiaztehsttehmepewraatteurrbeawtahstemmapienrtaatiunreedraetcothredisnpgecfiofripedHle1v.e2l, 4.0, 7.0, = 0.1C. and 9.0. Inall TonalbylepH1o2 fsubmumfaferriszewsasremsuelatssuorfetdheonpDHamyea0,sunortemtehnetpsHfoofrspaHmp1l.2e,s4.c0o,nt7.a0i,nianngdt9h.e0.teIsntasduvbesrttaennctley,, wTahse amtetahseurspeedcipfHioedfnDomaiyn0albulfefveelrswaitnhd tDhaeydeSvireaptliiocnatneotsaemxpcleesedfiorn=gal0.o1.fthe investigated pHs CONCLUSIONS `atTh5e0hyCdraonldytpicHst1a.b2ialtit3y7ofCH-fo2r4f6i1ve6dwaayss.teTstheed filnubourfifneatreedd aacqtuieveouisngsroelduiteinotnsofatHp-H2446.10,67w.a0,s9.0 Htdi-eam2o>4no6sn1te6rayaettaerpd.HtoT1hb.e2edahanytddarpoolHbytt4ia.ci0an-lepldhytfhsoatrlaabhltyeedrabotulfpyfHteircs9.si0ts,abnpiolHtitc7yo.o0nf,clatuhnsedivfpel.Huo4rT.ihn0eactidetidrfaaftceetribevunefcfeioenrfgrwreiedtcihoevnetroiefs --~ fTohrefdlautoraiinnadtiecdataecstitvheatinagrseigdniiefnitcabnettpwoeretnioDnaoyf0flaunodriDnaatyed5asctaimvpeleinsgwreadsileanrtgestratyhsadnis1s0olpveerdceanftt.er PadHdi4t.i0o-npohfthaacleattoenibturfifelre.toAtchceusraamtpelqeusantthiattatwieorneofetxhepdoistssooeltevdsetdcfornadcititoinoonfs finlupoHrin1a.t2eadnadctive. ingredient was not achievable for acetonitrile extract. Therefore, accurate mass balance of efxlturoarcitnsatfeodr atchteisveepiHnsgreddiidennottcoinudlidcantoetabneyacchhainevgeeds.toThthee flFuoNriMnaRtesdpaeccttirvaeofitnhgreedaiceenttonmiattreirlieal. cTflohuneocrmeiennaartsueuidroaencdsticvo oegnncgernedtirgR agttionoamnateripalit edid bnyotrgpdfoiircpatHge4.a0ndny hydrpoHly2t.i0cesbdueefgftrearidsna.ttihTeohnteoofoktbhseerved swuhbiscthanccaen(bHe-2a4c6c1o6u)n.tedFofrorpbHy1e.s2 iandg9.0arn ienggcease:ofthin the tes ncentration beyond that Substance was observed. It wfoarspeHst1i.m2ataendd tphHat 9t.hi0sbiunfcfreresa,serewsopeucltdivreleyp.reset hydrolysis of 0.06 and 0.03% the test substance. --~ - Company Sanitzed. Doss not contain Tara mm StHH2yAd6e r16o:lSSutbbiltyooffHi-62i4G61u6saasFaunFucntctiioonoffpplh____ --~ DuDupPoomnti1i3o97n0o RECORDS AND SAMPLE STORAGE NSepweacrikm,enDse(liafwaaprpel,icoarblaet),IrroanwModautnat,aainndRtehceofridnsalMraenpaorgtewmielnltb,eWrieltmaiinnegdtaotn,HaDseklealwlaLraeb.oratory, REFERENCES 1. OGurigdaenliisnaetsiofnorfoTresEtcionngoomfiCchCeomoipcearlast,iHoyndaronldyDsiesvealsoapFmuennctti(oOnEoCfD)p.H,MOaEyC1D2,G1u9i8d1e.linOeECD No. 111 --~ ee --immmctteeem Company Saniized: Does not contain TSCA Car SHHyedzreaool1yt6eic Stmability ofH-24W6O1m6aafsunFcuntcitioonoofpi --~ DDuubPeonmti-1s3o97n0 TABLES ~~ --~ -Y Company Sanitized. Doss notcontTSaCAiCnEL wa tamed _-- _ Table 1: Analytical results for aqueous stability of H-24616, pH 1.2, 37C, Days 0 (12-Feb-2004) and 5 (17-Feb-2004). . ETI mmm,|te . F C o r e e s r 7 ee ] r ] [aTole Ts e -- =w 1 e r] an| T Ce e -- ] -- Fe] [aTsTT meTwa [wr s ~ TE= 11 [wee Tw ? recovered in the acetonitrile extract ?sampleofacetonitrile extract not saved * represents total recovery from THFand acetonitrile extract * represents nominal concentrationoffluorinated active ingredient in the buffer ~ py AY 2iy4d6r1ol6y:tc Sabilty ofH-24616 as 3 Function ofp DuPont 13970 ~ Table 2: A5n0alCy,tiDcaaylsr0es(u5l-tsFefbo-r2a0q04u)eoaunsds5ta(b1il0i-tFyeobf-2H0-0244)6.16, pH 4.0 phthalate buffer, NnomaarniCoone1r0r1s6in|| MiesmierdeCooydpfgReknAnrIatSon| 34 Recovery feo o rrse7 Te1 rT m [Feeo T ToemoT maTw w T 1w | [eos TwTw [hme TT ] [eb Ty [[eoeoTTssT TToa mT1 ossmoewrs [| | we | [ooTs IeToone oT sm Tse | we [wr | 1 ~ [eeTw Tecovered Tnth TIF exact IE *rerperceosveenrtesd itottahleraecceotvoenrityrifloemextTrHacFt and acsonile extract represents nomial concentrationoffucrinatedscive ingredient nthe buffer ~ -_-- Company Saniized, Does not contain TSCA Cs Bt2de46s1es6:a woofo2-4t615ma3rFmoeinmftt opwuepn -- Table3: Analytical results for aqueous stability of H-24616, PH 4.0 citrate buffer, 50C, Days 0 (5-Feb-2004) and 5 (10-Feb-2004). [oonp|MmTtn,|| oSEmiEiti,| C[ r oorlr a o w 1 ses w || Coeooo T w ww T] ] =-- ----1] = owTT]1] Co o T a www w weT |w v] ] oe ew 1 w ] ] ~ TET [t= ov |] ] recovered in the acetonitrile extract ?the vial most likely leaked during the experiment "Day 5, replicate 2 excluded from calculation # represents nominal concentrationoffluorinated active ingredient in the buffer _ _-- companySeized, Dossnt continTSCA Gn HenyadseeesSwieteort24e1n6e3aranim oi wpmoarn -- Tabled: Analytical results for aqueous stability of H-24616, pH 7.0, 50C, Days 0 (29-Jan-2004) and 5 (03-Feb-2004). pr-- EE. hy mg) iS r i ooe m r w wr w T Tr ] [Coo TooolTleeT w T T ] C rCoT o wT e w iiaT ] ooT Tw wT w] ] EE e = T = ompanySanitized. Doesnotcontain Tsca rm) wHaostlley:yeSuubbiily of F-2i4G6i166m1aFFuurteoinonoofptipt puDuproowni1a35o7n0 --~ Table 5: Analytical results for aqueous stability of H-24616, pH 9.0, 50C, Days 0 (29-Jan-2004) and 5(03-Feb-2004). Re| orbiCosn|| MeoEas | nn Er e rw 1 T ooe Ts oon wo w m Tw] ] CooToTsn wT] F------ I ETT SeTw] C C sE we w mw T T] ] CITT wT] ~ I Te Roma nee of RT = I ee grein ---- --_ -_---- Company Sanizad. Doss not contain Tsca car HyHHdayo2d4rl6oy1li6tecSSaubiiltyooff H#2G4616a1saFuotnocniafio.nofpt__ DpuwProenmts1o35n7n0 -- Table 6: 5Me(a1s7-uFreebd-2c0o0n4c)e.ntrations ofl. PH 12, 37C, Days 0 (12-Feb-2004) and |e Jonecolonen]d| conan EN | i. rrr [ee Tw [ee Tw [eosme |] [lol 1 rr 71 TT ewe ow [1Tse|sme[ i| e rT [T Tmo Tw d Ta w | [ETmo e 1 Te ww aw l| [EloTeea 7m Tw |w | rrr T TT T SenietwenT [wwo || --_ rrr a a Tn |Te [essTT a es Teee rr TT New ew 1] TT somioeinin|a0] T 7 r 1] | 5[ Toc]ww Tee |ws] TT|EE A [| Tew wm ] CTT SiT vioiin | 05] _ : Ep cee pomrereoTSORS JHyaspytedSiubeof2s4a61b10 iPriionygto. fsicuatusiooDnwroeosms --_ Table 7: Measured concentrations of{QAYMor pH 4.0, 50C, pF 4.0 phthalate buffer, 50C, Days 0 (5-Feb-2004) and 5(10-Feb-2004). a [oneno]|chmen]clin[oer ee [[foeoT ToT]T o w ]1] [oe Tosa] [ (e re r 1] TT ew [wtT iooin| T ] rrr 7 [| woTe [T To T e w T s w | rrr TTTw] --_~ rr Sotr o [5] ese] [Ts ese TT Core TT [| mew| mw][1] TT sotto [0 rr11 oTTel Tm Tm [mT wm ww] [oeTsTems m T TTTew] CT Semin[0] -Y YQ Company Saiized Doss ne contin TSCAC: IdF--lyeSSubuibiyof2l 4: s6e16s3atFimctioonnoitt pDwerenaomn ~~ Table 8: Measured concentrations oor PH 4.0, 50C, pH 4.0 citrate buffer, 50C, Days 0 (5-Feb-2004) and 5 (10-Feb-2004). | oon[mn| cli ove | e e rer oe rT T r w w r| C E Er or T r T C oem e |T o aT n C eofw o w r wr ] [[moeelfee]lw ww w T1ww]] Crrrw 1 w ] ] Cr sews a] ~ E( EoI yo A e--m -- e (o ww ] Cr 1 C T[e S|r m0o ] [(oollee]w mw Te w w] ] [IorE lre ww S w| CT ewT e|] --_ EEE Company Sanized. Di oss roti contait n TSCAmaG--1 HFraosltiec Sail ofF-34616 1. Function p. tt. Dupon-13970 ~ Table: Measured concentrationsof QE pH 7.0, 50C, Days 0 (25-Jan-2004) and LLee] 5 (03-Feb-2004). rTrT 1 [ooTovTmT e 11 [roTo = 1 aw 1 1 [ro lel os 17 ~~ 1 =e T- 1 [ole 7 -- TT ww T -1 rrrr T Twee[a7 w [| [s=s 1 1 1 1"1 1 [ro[oloo|"vw | ew | mm | [ro[ele 1 m0| ww Tw| [e Tw l 1 e wn Ts w| TT 1 1 wes | | ~ 1 sovtioin|05 | Cr 11 [C +1o 1 s sw [ [ro [= 1 sw 17] [osTT ww [- | [os 1 1 mw 1 | r1r 1 1 Tae 1 sw 1 ss [= 7] 1TT 1 [ro[sea me ww Tw| [oo[sea Two | ww | ws | [Cos esTow 1 ww Tw] rT 1 1 awww] NoT r iond tT h clTat1 on o veags|sod aSnwdsaioGmoiisinn| 30 | --_ - ">mpany Sanitized. Doss notcontain TSCA CBI ~ IpSauebse tstttiesto Table 10: Measured concentrations of 5 (03-Feb-2003). oJws--in for pH 9.0, 50C, Days 0 (29-Jan-2004) and o f: d fed -- Cr ooeT T T T a ] C H Cre TT E [TEE IE C C S E oe es T T e e e s n] ] ] Cr ~ C H C r Te Tm To] C rrr T r w C e C o r T s 7 -------- CH o TeEeT Ts E T eo] ] Cr Cre T T r T s= e r ] ] CrTTT oe] ~ a CompanSantas, besetc son cn woied Sui of124616 2 Fsinot -- Table 11: Dailytemperature readings of the shaking water bath. Dupes r C EE om r m C Tm m aTTooe] Fe e e Temme m TTaoa]] o a T wwmea a] aT omem T Tma] aToomma aTnw]] ~ T T EE E e T v-- e T-- w aor] Co o Te Ta emm a] a [-- e Tw] Tn eeTTae] Co T m a mm a aaee] 3 5 wren |soo] --_ a Company Sanitizeq, Donote contsain Tsca gy ri HHe y2d4r6o1lG 6m y:tc Sty ai bilityoG offH-24e 616 asa a Funcl tionofu pH nctonofDDip uProontm1t 3a9s70n Ps `Table 12: pH measurements for Day 0 and Day 5. -- Tr r r FeTose emprefs oe s Tw] Te Ls sw Ls Te 1 7 ---- [a Tr r se Tw r 5 s : +0 [as] FsT T sTT w aTww seTw] I LoThTe aes5 e ---- 17 -- ssT Tw e Tw aw] --~ f Te aw] Taw] ee ------rrr TT oe 5 --r 7 I s SE E e MTS STNe TN FsTT sT o e w ow] oT1 7 ---- 1 eeTo 5 17 Tw Te TT Tes L LPHs s mot measT urT ed orreplicate sampT les; mese T ss 5 ured buffr pH weo os 1.s w20 ] ] 22pppHHH 44n.o.0t0 mcpieatarlastauertbeeudffbfeuorfrfreerplicate samples; measured buffer pH was 3.99 -- 7pppHHH nnnooottt mmmeeeaaasssuuurrreeeddd ffrooerrplrrieecppalltiieccaasttaeemspsalaemmspp;lleemsse;;ammseeuaarsseuudrrbeeuddffbbeuurffffpeeirrl ppwHHasww4ia.ss0169..9090 CoRan Sanitized. Doss not contin rsa car H2y4d6ro1l6y:tc Stability ofH-24616as Functionof pH --~ DuPont 13970 -- FIGURES eee. `Company Sanilized. Doesnotcontain TSCA CEI H2Hudy6rdo1rl6yoileSSatabbiillityy ooffH-i24661l66uassaFuFnucnitoonn.oafippi wDuopomn1e39n70 ~~ Figure 1: cEaxlaimbrpalteionFsNtaMnRdarsdp(eAc)t,rafloufor1i0n5a0temdga/cLtifvleuionrginraetdeidenatctrievceovienrgerdediinenTtHF extract (B), and acetonitrile extract (C) for Day 5, pH 4.0 phthalate buffer. | BES _ E i a" e ~ B| ] | [MIl | -- A .| i er Hl| | I rma |prersessesemenrssmmirseLnemOa? ROS E a i N --~ -_-- Company Sanitized. Doss not contain TSCA Car HHiarsstle Sabo 24616 Fnton fi Dupent13970 ~~ Figure 2: cEaxlaimbrpalteionFstNaMndRarsdp(eAc)t,rafolufor2i1n0atmegd/aLctfilvueoirningarteeddieancttirveecionvgerreeddiienntTHF extract (B), and acetonitrile extract (C) for Day 5, pH 1.2 buffer. } A | | creme 3 a LW ohnI e 11-- -| || r_ -- |B | il I N I M Jl | Ll a tsetse, wienind Apdt E is ee --~ Company Sanitized. Does not contain TSCA CBI roma : te A WilAN ~| |B || EEE | r2Hy4d6re1ol6y:toc StarbilittyofoH-24y616oas alFunctlioniofpp.idolmatuctioDDunuobomona1s3o9l7n0 ~ Figured: bTEuxHfafEmeprexl(tBer)a."c"tF NfoMr DRasyp0e,ctprHa o4.f0thcietfrlatueorbiunfafteerd(aAc)tiavnedinDgaryed5i,epntHr4e.c0ovceitrreadtein i | -- LJ UL i Ed we ---------- ~ | BE t Ye Ee Wo _ ~~ EompanySanitized.Dossnotcontain TSCACB aHyddsrloy:tc Stability ofH.24616 as a Futonofpit DuPon13970 ~ Figure 5: Example 'F THF extract fNorMDRasyp0e,ctprHao7f.t0hbeufffleuror(i4n)ataenddaDcatiyve5,inpgHre7d.i0enbtufrfeecrov(Be)r.ed in dork || | J I 'vse L1 | (SLU i ~~ |& --_-- | Ld a J|| U| o gee sn --_ - " `Company Sanitized.DoesnolcontainTSCACB DHhyauosltyebi:coSisltyooff2f4i6c1a6abFweiimoonoplis.h oDuwPonetr13570 ~~ Figure 6: Example 'F THE extract NMR spectraofthe fluorinated for Day 0, pH 9.0 buffer (4) and active ingredient recovered Day 5, pH 9.0 buffer (B). in I ' Hl || Ili 1 | WPL. | I| N: E B Ef | |I nm ! IL --_ Sompany Sanitizsd. Doss not contain TSA Ga HHaosltiec SailofF-246136 Fcion ofpit. = Fiiguregure77:: Representativetivecalibcalribaratiionon curcuvrve fogmp)\ aoolrrbemripaaotiiuoonnndccruoarvemef:ic1e.n0t21r9=10.x99+9760140,74628= 0.999208 eusrpeopnsee:tyLpien:eanr,teOrmiagiln:StEdx(clRuedfe2,)W,eAirgehai"ng(:1S1 Conoc.e1rSanAsr:eNa)ano 10539- : Duran 13970 Response- | 0 2000 oon | Goo Teo romeooo) wm Figur8e: Representatiys chroma amsof102 Qumran sandra 52100475 Smo 10) 100 % (a3m61e3 4p08r22s34 SRoTS GamaES. 249466034,050253 021o003015 Smoove) on |% Comme 212210334307008 SIR :015cnannelm5is.n stlrora.5i2n0 o. T-o Ihe ak a: k ah min --- Gompany Sanitzed. Dossnotcoiain TSCACBL 2Hy4d6io1l6y:ic Silty of H-24616 as Functionofpi DuPont 13570 ~ Figure 9: Representative chromatograms stability sample and 520 pg for Day pH 4.0 citrate buffer hydrolytic ternal standard. 021004538 Smooth(vin,13) Too % Cwenn 117335367.03 679545 SR 015 chann4e6l3s,5263685004005 | o 215 021004639 Smooth(vn, 1:3) To 0 *% 15:385-8278A_ 21224886147.2954 SIR ofs channels m5i-n 1.2486870.+502086 o 100 200 3.00 400 500 min| ~ Figure 10: Representatigg chromagggrams for Day 5, pH 4.0 citrate buffer blank sample 021004507 Smooth(in 13) #5 205 w ati2.49 SIR 458 of chann4e8l3s,5253 43 NS sstreion 362 021o004507 Smooth(hn 1; x3) Too % Cr S-- 21034588748 1213495 SIR of channels min 5- 122416274,050268 a : 100 200 500 400 500 mo] -- - `Company Sanitized. Doss not contain TSCA CBI