Document 71mXjzR1KQzBRK7j6qjdjgg4a
MR 2155S
ARR226 -- 1335
REPORT
96-HOUR ACUTE TOXICITY STUDY IN ZEBRA-FISH WITH T-4127
(STATIC)
NONTOOTXOXSePnreotject 33i4a33e8t7
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NOTOK BY. `Harogenassc, Th Nahrnds
Io sty esrndin is ooabort portedandwas conto compare
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ne Uns Stes Foodnd Ong Acris. 2 Codeof Fara Raion Par 55.
TIhrUenet SStonEvhanmntal rts Agecy (FFA) Tita40 Ce of Fora
TS oe Unkn Sto EnianmatalPrtoctin Agar TSCA) To 4 Code of Fda
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Tanz QUALITY ASSURANCE STATEMENT
NOTOX Project 334358
NOTOX BV, sHertogenbosoh, The Netheriands
rTehsiusltrsepaocrcturwaatselayudreiftleedctbiytehaewNOaTtaO.X Quality Assurance Unit to ensure tht the methods and TDuhroindgatheesoonf QsuialinsApsescutrioannsceprioncsepdeucrtieosnsapapnlidcaabuleditoahriosipoenobfesltou.dy were inspected.
DATESOF QAU INSPECTIONSIAUDITS onsite inspecion(s) (Process) JMayy01931:53,1,22000022 ((EAcnoatlyotxiiccao!loSguyp)por) protocol inspecion(s) (Such)
December 06,2001
REPORTING DATES JJuyne1074,,22000022 December 06, 2001
report audit(s) (Study)
Aor 2629,2003
Apri 29,200
Head of Qulty Assurance
(ome
ee ystems
Ca
gu
Tater
NOTOX Project 334338
sowwary 96.Hour Acute Torcy Study in Zebrafish wih T4127. oTTheyostuodrytparvocaedaGuvnraerstnidoneasOciEroCinbDeodfgunedcoosleixnirgcetNpyoo,.rtP2wu0be3lr:iocFabitisaohsneANcoo.unteLt3hT8a3x,EcEDtCeycdTeiemrsebctet,irvAe1d99o92p26t,6e:dC1P1.a7r"JtACcy:ute To52. TT7r4oo1m2aau7nbyaittbaaensneasskywpcahotnoessripsShtionngtooafmP(eocfraltTiu4oon1rioc2o7pcatwraa.nseBsduaeftsoeenrymioimnneeitdhhe0ylraebmsoipbdoeonts(weaenoeifonnti0ec5paaanrino)dnia0cn.6dpamrtaofahte .
19.52 06C.
"tnTehoeesratayyssstoAaortanerd,iwiocthoatnmypeirsotwpaaessrteiseatxopaoccsliiunndggesdfo.avtAeennsaliofyssh1i0sp0oafrmtcghoencasneadnmrpbalotetoshnfaaogknaeenftcicutreairlonydg ahsniedrmeadinlfoerst C7hm0oc1awrnsihrieaaittonioncsarcavlseaortuaetgdoenaevxaepnrods1u0r.2e586cohGnoc1uetnitersahtteipounanicioatdlrweeaadstes0od.l5vt1oenmr.ltTeihnrstathve2fr4aehgroeeuedrxsspoaolsusutiro0en.7a5nd i h6o7rsomdonlolelunrieohmna,iunwinhnigif1ra0en1dypsroilaouuot,noH.faoBnuvanotghdiieboorntsghti2e4uoenuflfrfescrteosfdcxospnofuismunerdeaaanldhifngihsoheordiraecedtuiianlhhodied Concentration in he lord sain. aAWfnaittnaeelrrtieAsotcr3wcaammsampsoeidranftoieromdfedF3reaxchpoiouosrnis.n(gWAAlslPol)vaepsnrsfewipsaahrrpoedeuacetodanclneonat1chriantgfiionrnaaltteeso4ft51w,0a10s0,gp2rf0e,-ao4n5sdtaaobndldaw1ni0hk0% of CGaemmssotttaaconydemwvoaaptyosrptaatainrocnc.weeAsThSoiiultgathopvohaspavobeulslespclra.eussSseuarmoeprlwedaassctrtaelkaetsnevulorfyitnoesgwt,ncoebnacnseneltdraeotnsitotnhwse.erHTeehnaerdryedfeodre rol t sopropanol ai a ral of 11510 prevent any action! oss. SCTiohnercoaecnnaaltuyatttiioocannlssptsraooukglerdnantmootbparbesaveemdnatonniaadmisencaersdpuiatrnem1omregenaohtsfastnhaen6da0%vnoliaotafiltipzeaarttinonsa.hloTwhceoedncmteehnaattsraurtreieodna,ctiunalsite of CHiognheeasntrcaotnotnesntcroaounsnoahbshqaueanretloevfaonthora dWehteorlmiensattipoenroifodthweihLCixh.e xcopion of fhewo anntdheercmopnatrroaltunro)morroamlaitiynweawsionbisetrnveerd.anFguersthperre,saclrlitbaetd bcoyndnietipornosto(cpoHl., oxygen conceniration 1fAa0iw.1atTwehhriespnchoabrsareseeosfdpoTon-nd4ie1d2e7waoirnigo0i.nn7at6piangrgtifofmiornoa2w4laohatodueirnpsgha(rasiteme.iofNteo1s0)m0aomnrdga,0l4io4ndmu1oc6el5sdoh1ra04n08%5hm0oou%rrtsmaloi(rtianyalillny royceSaEineaLt aCovofersagee cronacaonxtpaotseends1ofT-40013257mwgas(<ina1lmiofs)(abnodw0e.1n50m.1o6 ainmd 0.e6s4).gHhencbea,sed on quantification of ha aio par. haLirmaeiutrrinagas tnTedharrneaolystaaarllesauhplapionrpthooossnseiobnollfyytththeheaaanhnioioonancptauarar.ltcHoofonwc-ee4nv1te2rra7,t,ihovenasslopfointsfhorcthaietdienostntcpgaor7lt9dweenrWtietbhe nreesurts rosa and deci1d acoopt he hgh 1x abating (RSD) based on the current
s
Tar27
NOTOX Project 334338
PREFACE Sponsor
Study Coordinator Study Monitor
Testing Faciity `SAtquudatyiDciTrocxtiocrology: TAenaclhyntiiccaall Ccohoermdiisntartyo.r Principal Scientist Study Plan
33MM CCeonrtpeorr,atBeuiTdoixnigco2l0og-y26-02 SP.TO..PBAoUxL,33M2I2N0NESOTA 55133-3220 USA. Mrs. M. Mitchell 3MrMs.EnOrv.irS.onBmeenatcahl Technology and Safety Services S9T3.5 PBAuUsLh,AMveInNuNe,ESBuOiTldAin5g512-434E-09 USA. NHOamTbOaXkBe.nVw.etering 7 5231 DD s-Hertogenbosch The Nethertands. IDnrgs.. EM..JHB.ogeMrust.saards Ir. MJ. Brekelmans SSttaarrtt ooff pfrrsotjeecxt:poDseurcee:mbJuelry 0051,, 22000012 Completion of ast exposure: November 15, 2002 Completion of analysis: November 15, 2002 Completion of project: May 09, 2003
TEST SUBSTANCE
DeIsgcerntiipftciatoinon CBoamtpcohsition Test substance storage Stabilty under storage conditions. SEtxapbiirlytydaitnewater
TDa4r1k2a7mber waxy solid 9D5-2-49919%lotF2luoroelastomer curative
<2% N-Methyl Perfluorooctanesuiphonamide <<11%% EMtehtyhlylAlAclochoohlol At ro<o1m%teImspoeprraotpyulreAlincothhoeldark S3t1abMlae.rch 2003 Not indicated
"The sponsor is responsible for all est substance data unless determined by NOTOX.
Ta127
NOTOX Project 334338
PURPOSE e"fTfheecptsuripnoDsaeniofortehreiosdtuurdiynigs taoneveaxlpuoastuerethpeetreisotdsoufb9s6tahnocuersfoarnids,aifbipolsysitbolgee,nteordaetteearcmuitneettohxiec: LCuyat al observation times.
GUIDELINES "mTehtehsotdusdyfoprrtohceedduerteersmdienastciroinbeodf iencotthoixsirceiptyo,rtPwubelriecabtaisonedNoo.nLt3h8e3,EEDCecdeirmebcetirve199922/6,9C;.1P.ar"tAcC:ute toxic for fish", and the OECD guideline No. 203: "Fish Acute Toxicly Test", Adopted 17 July. 1992.
DEFINITIONS pFiesdhunwcelreeacnodnsviidsiebrleedbtroeabtehidnegamdowvheemnenntosraeracetaibosnenwta.s observed alter touching the caudal The LCs is the concentration kiling 50% of the fish after a defined period of exposure.
TEST SYSTEM Specios Source
Mean length
Mean weight
Characteristics Reason for selection
Total fish used
Zebrafish (Danio rerio, Teleostei, Cyprinidae). `NOetmhaemrelnantdasl.Fish Hatchery Atlanta, Hellevoetsius, the: FLiinmailt tteesstt:: 2257 0022 ccmm LiFminiatltteosstt:: 00234640.00.812gg Healthy fish supplied with a health certificate. a"Tchcisepstyesdtesmpehciaess.been selected as an intemationally 6
Taz
NOTOX Project 334338
HOLDING Quarantine/Acclmatsation Medium
Measurements Feeding Gontrol of sensitivity Valid of batch
Atleast 12 days ater elivery.
w1aSt0e-rmepudriiufime,d fboyrmruelvaetresde uossimnogsiMsi:llM-iRlloiwpoarteerCo(r1pa.p-
cBorcord, Mass.s,omUoSnA) withthe folowing composition:
nIoe
spmeo
o
14a5nmggt
sHooors "omneorn
:
Harnoss 250mg CaCO
cpoHn,ceinrtaratteioann:donncieeacwoenecke.ntTreatmipoenraatnudrea:mmevoenriyaday.
Day with Trouvi.
"aTphpeernedsueldtstoofthteherempoosrtt.recent performed test are
hInethseevbeatncdhaoyfsfpisrhiours0edtfhoe ttahrettoefstt,hmoorttesatliwtyasdulreisns.g than 5%.
PREPARATION OF TEST SOLUTIONS
dTihsesosltvaenddatersdt tseustbsptraoncceeducronecsernetqruaitrieodnsgeonresrtaatbiloen aonfdtehtomsoolguatinoonos,uswhmiicxhtucroenstaoirndGiospmeprlsaiaonlsy. sTuhbestteasntciengoonf tchoencweantterratsiurofnascot)h.at disturbed the tst system wero prevented (e.g. im of the test TTr4i1p2he7niyslabesnaztyicpohnsoisspthionngioufmP(ecraltioornoiaccptaatn.esBualsfoendyoimnetthheylraemsipdoens(eanoifotnihce paanrio)naicndpar of the t1e8s.t5s2ub0s6tanCc(eNwOatTeOrXsoPlruojbeicltiot3fy40T8-4421,2u7swiangstdheetceromliunmendefolbuteionbmeettwhooed)n.0.6 and 0.9 mg! at TLiwmoitsttotck solutions were prepared by weighing 500 mg of T-4127 on a glass surface (wie). oAbfitearinpalaclionagditnhgesreatceaorfri1er0s0imngm].easBuortihnmgixfltausrkess5weirre mofagtneesttimcealdliyusmtweads farddtehdreteodeaaysc.h Aflfatsekr to hatethsetirbroitngtopmeroifotdhthfelmasikxst.urOensewoefrtehcelmoairxtbuurtescownatsaiunseded(oasst ssuubcshtwanhcie ptahretiscleecso,nwdhimcihxtsuerteiweads sfiutbosrtoadntcheropuagrthicalpeasp(ecar. f>ie5rum()S.chTishichfianralatnedstScsholuustlilon60w4a)stcolreearmoavnedtchoeloluarrlgeesrs.undissolved ost
Tat27
NOTOX Project 334338
AFlilnagllatessstware used in the final test was pre-treated with dichiorodmethyisilans to prevent loss of 5te0s0t smugbsotfaTn-c4e1d2u7eonfoaadgsloarspstsiuornfatcoeg.laAsftsesruprlfaacciensg.tAhestcoacrrkiseorliuntiaonmewaassurpirnegpafrleasdkb5ytwreeigofhitnegst f`lmaesdkiwuamswaprseapadrdeedditnotohbetasianmaelwoaaydinbgutrawtiethoofut10a0ddmign.g tSesitmsutuabnsetoaunscley(,blaannokt-hcoenrtrmoe)a.suArfienrgthe `stsierprairnagtpieornifoudn,ntehleasnodlulteitontoprsetapbairleisdeaftora2o%adhionurgs.ratSeuobfse1q0u0enmtgl/y twhaesWtartaensrfAercrceodmtomoadated cFornaccteinotnra(tWiAonFs)wwearsesperpeapraarteeddbfyrdoimlutthieoncoefntthreoWfAtFh.e sAelplafriantaiotenstfusnonleult.ioLnoswweerrteesctlear and colourless. Part of the test solutions prepared were used for testing with Daphnia magna (NOTOX Project 334349).
uMIT TEST
SAolluimtiitotnepstrewpaasrepderatfoarmloeaddienxgporsaitengofs1e0v0enmfgifshapnedr caonblcaennkt-rcaotnitornolt.o a fitered and an unfitered
`Sampling: VForlequumeency Storage
a1t0tm=i0hf,ro1m=2t4hehaapnpdro1x=i9m6ath,e centre of the test vessel. oNfotsaamppplliincga.ble, all samples were analysed on the day
FINAL TEST: TEST CONCENTRATIONS
T4127
45,10,20, 45 and 100% ofaWAF prepared ata
loading rate of 100 mol.
Control
Tost medium without test substance or other additives
(Biank-contro).
TEST PROCEDURE AND CONDITIONS
Test duration
96 hours
Test type
Static without interim renewal of test solutions.
Test vessels
d1i0cihrieorso,diamlle-tghlyalsss.itaPnree.-reated with Although vapour pressure was relatively low, based on the Henry constant evaporation was stil a possible tchaeussetufdoyr wdaecsrpeearsfeoorfmteedstincoanirc-etnitgrhtatvieosnsse.lsT.herefore
Test medium
c1oSn0c-emnetdraituimo,naheardatreedacunhteildtshaetudriastsioolnvaedndoxtyhegepnH had stabilised. After aeration the hardness was 250 mg CaCO; per lire and the pH was 8.0.
Tat27
NOTOX Project 334338
Number of fish Loading amination Aeration Feeding
Introduction of fish Euthanasia
7fsh per concentration and control 028 fish, .7fish por ives of test medium. 16 hours photoperiod daly The test moda were not aerated curing the test. tNhoefteoteadlitnegstfrpoermio4d8.hours pir to th test and during Within 1% hours after preparation of the test media AKtitlheeboynedxopfostihnegestthetmheosucrav.iv1i.n2g%fiesthhwyolreneergaypicdolly monophenylthar in water.
SAMPLING FOR ANALYSIS OF TEST CONCENTRATIONS
sDuppolciiccataetosnamopflhees woreirgeintoafkesnamfprloemsthteaksetnocukrsinogothne tdeisrte.ctDluyraifntgrhperefpianralattieosnt odfuptlhiecaWteAF as a Soafmepxlpeossuwreer.oAttakheenefnrdoom atllhecotnecsetnpterraitoidondsupacndatheesbamipalkecsonwleorloataktehne fstoarmt aalnddautfnrs48(4h.o5u,rs d0e,sc2r0ibaenddin45th%e)aopfptehnadWeAdFAnaalnydtitchaelbRleapnoir-tcontol for analysis. The method of analysis is
Sampling: TVroelautmmeent Storage
5Sammipfiroosmwtohreeapapcrdoexdimdiarteectcleynttoreisoofptrohpatneoslt avtesaserlsa.to oNfot1:ap5p0lipcrabolvee,ntsaamnpylaodsdiwteiornealanlaoslsy.sed on tha day of
`sampling.
MEASUREMENTS AND RECORDINGS
Mortalty and other ffects
Aex1p2o4s,ur2e4.,4D8e,ad72fiashndwe9r6ehrouermsovfeoldlowwhiengnthobsestravretd.of
Fish length and weight
aTnedn mfiesahsoufrtehde bpraitocrhtoutshae fstoarrtthoefsthttwesetr.s weighed
Dissolved oxygen conten,
Daily in all vessels, beginning at the start of the tt
PH and temperature.
(day 0).
DATA HANDLING Determination of the average exposure concentrations The average exposure concsniraions ove th fist 26 o 48 hours of exposure were calculated 25 [Cg XC... . boing tho geometric means of tha concoriratons of T-127 measured in the
samples taken at the start (Cy.o) and after 24 or 48 hoursof exposure (Cu).
rar
----
20x[Cro Cr +72xCiz Crm, 48%YCrgXCoca +48. [Cos Coe
ag gama f bo crcaratora e T4127 mosses args en the start (Cio), after 24 or 48 hours (Cuuz0a Ciuds) and the end of the test (Ciuse)-
ims
Wesnsoscmcnsans
rersasrsboshsmowd sieepete
at
ie a
massates moro
eraser hos reratdtroi Ta
Dtz g of
posts
dad
Sper
essot shir so
a
-er [oo [am [an[om[ow| p=e
["Bankeowa [7 | 0| 0[0|oIo[ o |
[roomed | 7 | 0 | 7| 7 | 7[7| wo |
SE TmREeEEA rs [roountmered | 7 |@ | | i| | | to |
------ eo mss
gt xn cons tresosneTsl.rhni.
ara
NOTOX Project 334038
Tablo 2 p-values anddissolved oxygen concentrations (mg/) during the fmt est. Tosangraie | owo|buyi |_ bwz| ows |owe |
[[SSn0oe0ouoinmtneosroesas1--||587010[[|sassa+|[[777587[[|8oa5o[||7r7o37[[[7s1ss||77.75T[|76.s7T||77T57[[78.54]|} Table: Temperatures (1C) moasured dung the mi tsi.
TesTtiangrate
enene [toocamered |
ses[|meee | maer|e.s|. ms]|
[opened| zat | ete | ma | ms | ms |
Finaltest:
Measured concenraions TSohpornecseudsAonfayaniaclaylsiRseopfortht sample taken during the sty are described in Table 8 of he tEDu1or0irn8gsmtahvheircnsoanaafdnsdietdnhgteo43e3x3.p1mois1ua0ro0ep%rcoopoahnonsl aasolndfostaarnaoaacnyhdgzoee,dofoSdianumtpotlnnes AooffPIaSwm0alDttLeaCkaeMcnStfiNooSnm(phrreiesepceairoend omvoglr.umTtehie10a0.ul0l)y1. RimeggchAov(ceo1rn1cy7es%natarmanptdlieo1sn1wp1er%orvereepdsrpteeopcabireveebdne)il.noIwSLOO-Dm.edRieucmovaetr0i.e0s1amtg0/.l1, m0.g1/mga/nlda0n.d6 m0.g6 e`cSsaimpeolresewneprinocaa0tkeo7.nTnihn5eprloasotfsoalalfprrrootmcotahduetotnnosTtaasinoldoutm4i.eoaNnssoutaranedhsaettaaicknehn,pdreetcrreeaatseed isnacmpalnecewnarsaton
Tar
NOTOX Project 334338
Table 4 Meanof duplicate samples and ther deviation.
otoroamwEgTar
Measured concentration (maf) on
eancomncoenntrator
[4s 0
|<0 [| oom | oo | om
| oom | oo
[| | ss |
2 [es
| oe | ois | ows [v2 1 02s | 00 | oze | 12
| |
[oo 1 1
oss | os | oss
an
| Mem
|2 | | Ooviaton |
[a [wo
1 |
=wo [<o [-- <wo [<0 [--
--T-- --T--
--} --
.
20 4s
| oo | oom 1 oom | oom
| oon | mn | oo |
| |
[io | om | os
1
sen
| osx| s _ | Mean | pevition |
as wo
|<0 |<0
[|[ woo 1| | coo [1]
2 [as
|<0 | co[11 1 oo | oo | oo [| a |
heDuephleixicaortnegssnaicmaoplnldecesacnsrthaaoaoswneeosdfaosftihgdenui1pf6cisacalcnostncsdeainmftperlraeetsnicovensrinwscishohndmoicnaeirncFaiilrgusarlateopi11eo-si0nn.ocrlAuttdhtee=s4ct8ohnehccaeunutrsreaastnidfoonrast
Measured n45 and 100%of ho wate ration
Figur 1: Concentrationcurvesfor th thaohigheossttconcentations.
113607_334338_ZA
o 1.000 E p ESS SEy DeRE ir
2: a Sasa E
aGel
H EonE
|
E BrEeEEa el
se FEE
Time (hours.
x
|,
% esigenre: nS -
ay2orum:
rT m my n te crac Tae Tol so i ri ates hans
tong
a
pestcre
i nr ar aot
cotannn
1048
rra r son morar yaypo
[ow=oe[ [Fa w ww] [| ee
["Bamccowo | 7 | 0 | 0 |o |o |oIo |
[s[ es 7 7 1 1 w o ol lo oTo ol lT o oT l0 0 o oo | |
[ [2 Te --T7 7T To ol Io ol "o ol lo ol To ol T70 0 |
C aee Chet1 ec7 ta[do aI ge5s ,T717J7 |wo|
aa| |w omeue TE
LIE, en ni|S
z n
wo | a Le
|
Exmimcmirs
rT ates stmt Svos tc ngeogt otToe
"iooman [owe ]w we Tam | omo|bwi | ow:| bys | ows |
[Blankconva |80|87|74| 66 |75|66 | 76 |65| 75 |67|
[4s [so | 8s| 75|76|75675 76 7375]72]
[to
7s |8a [7s |77[75[74 | 76 [73[75|71|
[20 [76| 8a [74[74[75[71 |75s[66|75|62|
[4s 170|82 |75[63|72|57| 74| 65 |74| sa|
CoT7sl76[7260 -|-|-|-T-T-7]
T4127
NOTOX Project 334338
Table 8: Temperatures (C) measuredduring the final test.
ofTaataWraF (100 mg
["oamcoms | [as | [to [20 | [a |
Cow 1
220 218 218 ats ear ats
| ws | ma | wz | ws | | ze | mo | mo | ms | | ae | 20 | a2 | as | | ws | aa | er | as | | meTar | 2x | as | | we [T7777]
ACCEPTABILITY OF THE TEST 21.. ANlolmtoersttaclointdyiwtaiosnso(bpsHe,rvoexdygiennthceonccoentnrtorlatgiroonuapn.dtemperature) remained within the ranges 3. Tprheescarniablyetdicbayltphreopgrroatomcobla.sed on measurement of the anionic part showed that the actual
Scpointceenotfraalltipornescacuotuilodnsnottabkeenmtaoinptraeivneendt aatdsmoorrpteitohnatno8g0la%ssoafnhdevoilnaittiallizcaotinocne.ntTrhateion, in emxecaesputrioendocfotnhceatntwroathiiognhsesctouclodncneonttbreatqiuoannstiwfhiiecdhfoarrethreelwehvoalnet fteosrtdpeetreiromdinwaittihonthoef the LCie.
CONCLUSIONS
aUntdeeorf t1h0e0cmongd,itiionndsuocfedth1e0p0r%esmenotrateisttys ianwfaisthe.rTphihsasceororfeTs-p4o1n2d7edorwiigtihna0t.i7n6g fmrgomaforl2oa4dhiongurs p(lhiamistet.esNs)oamnordta0l.i4t4y morglafsosr4th8anho5u0r%s (mfoirnatlaltelsyt)ocwchuernrebdaasteadvoenratgheecaonnicoennitcrpaatritoninstohfe0w.a0t9e5rmg/l <(f1inaml tgest()baentdwe0e.n160.m16Aan(idmi0t.t4e4st)m.gH)enbcaes,edthoen9q6u-aLntCisf,icoatfizoenborfa-tfhieshaneixopnoicsepadrtt.o T-4127 was
eLinmsiutriendg.tThehearneailystiacarleasluisptpiocrptoossniboinlllyytthheatatnihoenaicctpuaarltcoofncTe-n4t1r2a7t,iovnalsiodfittyhoef ctahteiotneisct cpoarutldwenortebe
fhuirgthheerr arnedsemaorcrheasntdabdleectihdaendtthoosaeccoefptthtehaenhiiognhilcyptaortx.icHloawbeelvleirng, (tRhSe0s)pboanssoerd doind tnhoet caugrrreeentwih
resus.
gl
en
NoroxPs
-- sr nt ty
ve an FCP ROTOR Foe 5541 0
El scp densi i pr vt sds eEnCTdroivo5r88 PC
te iede ws carthe htte f ht inSednyFoe,
EA iCogrnA s: 050100S 15 022 rd
mgt
SOc
rn httn lr
C | | Sm [| a m Re Tr TTw| 1,
[owas
| 5 |o|o[o [0|o |o |
floss
Ts 1oo oToo[0 |
foo TsToToTo[0[0I"0]
fos 15[0IoI oo[0I"0 |
[loz
TsTo "oo ol] oo [0|
lR Cosuseteo ve 1 g5or[cm0 nest|on2rtIo4oIo1 a hse+]taJ
e rorno cts re psa LPeENTSAiCSOPC3PYcEo.aidnvsnmeereoioSaea ersoce8in
Ee ere bearrrre ors Pavers
The raw data and report from this study are en nn kept in the NOTOX archives. The test described
ANALYTICAL REPORT
.
96 HOUR ACUTE TOXICITY STUDY IN ZEBRA-FISH WITH T4127
(STATIC); DETERMINATION OF THE CONCENTRATIONS
NONTOOTXOXSuPbrsotjeacntce331413336807 -Page 10123
T4127 REPORT APPROVAL PRINCIPAL SCIENTIST:
NOTOX Project 334338
(IrA. nJai.ytCi.calBrCheekmeiismtarnys)
Al il Date: fy et, 2003
T4127
NOTOX Project 334338
PREFACE Analytical study
Start:
1 July 2002
Completed: 16 November 2002
PURPOSE The purpose of the analytical study was to determine the test concentrations and to validate the analytical methods used.
REAGENTS Mill-Q water
Methanol 1S0-medium Ammonium acetate 2.0 mM ammonium acetate Isopropanol (2-propanol) Internal standard solution
Isopropanol containing intemal
standard
Tap water purified by reversed osmosis and seuxbcsheaqnugeentclayrtpriadsgsees;d Moivlelirpaorcet,ivBaetdefdocrda,rbMoAn,aUndSAionP.a., Merck, Darmstadt, Germany see main report Fractopur, Merck, Darmstad, Germany. 154 mg ammonium acetate in 1 lire of Mill-Q water p.a., Merck, Darmstadt, Germany 100 ulof a solution of 1204 mg perfluorooctane sulfonate in methanol filed up to 10 mi with 50/50 (vv) 1S0-medium/methanol, 140 ulof a solutionof2616 mg perfluorooctane sisuofpornoapatneolin. methanol filed up to 2000 mi with
INTERNAL STANDARD
NIademnteification number Description PuBraittcyh number CeErxtpiifriyeDdate `SStupoprlaigeerconditions.
APeSr5f1ls7orooctane sulfonate (FC-95) `White solid (determined at NOTOX) 9L0o.t4291%7 Y21esFebruary 2003 (determined at NOTOX) AatMoom temperature in the dark
"The sponsor is responsible for all test substance data unless determined by NOTOX.
T4127
NOTOX Project 334338
SAMPLE PRETREATMENT PROCEDURE
Analyses in July 2002
2E0a0chulsianmtperlneal(1s0tamnld)awrdassoqluuatnitointwataisvelaydtdreadnstfoertrheedtsaomapl2e0smtiakveonluamtetr=i2c4falansdk.tAv=9o6lhuoumres,oaffter w1h2i0cuhght)h.e fNloasiknstewrenralesftialenddaurpdtwoatsheamdadrekd utsoitnhgemfeltashkasnocoln(tianitneimnagltshteatn=d0arsdamcpolnecse.ntration
Analyses in November 2002
`Iandodirtidoneatrlo ipnrfeovremnattifornosmupapdlsiorepdtiboyn,thiet wsaposnsdoerc,ideeadchtoscahamnege(s5maim)pwleasprter-atrnesaftemrernetd. Based on
.
q(iunatnetrintaaltisvtealnydianrtdo cao5n0cemnitrvaotliuomnet1r6ic4fgla)skwaansda2d5dmedi.of isopropanol containing internal standard
ANALYTICAL METHODS
TTr4i1p2h7eniyslbaesnazltylcponhsoisspthionngioufmP(ecraftliuonoircoopcatr)a.neUspulofnonryeiqmueetshtyolfamtihdees(paonnisoonri,c pqauratn)tiatnatdive analyses. `wwietrheoMnalsysbSapsecetdroomnetthreicandieotneicctipoanrt(oLfCMT-S4M1S2)7.uTshinegaHnialgyhtiPcearlfmoertmhaondceusLieqduiidn JCuhlryo2m0at0o2gwraasphy Tnhoetrseefnosriel,ivae seencooungdhmteotmheoadswuraescuosnecdenitnraNtoivoenmsbien rpr2e0t0r2e.ated samples fromthefina test.
Conditionsfor the analytical method used in July 2002
Column Mobile phase
Betasil C18, 50 x 2.0 mm; d,=5 ym (Thermo Hypersi GKreaydsiteonnte) EElluveennss AB:: 21000m%Mmaetmhmaonnoilum acetate
FCloolwumn temperature Autosampler temperature. DeItnjeeccttiioonnvolume
T4127, anionic part
Internal standard
[ooo "Teo= "Tio 1]
[oo T= eo o -- 7]
[sso
5s
Tes
[Jooe Ts
Tes
[Moo Too
Tio
1]
[30s0 wolimin Te0 o ]
`ambient temperature
`ambient temperature
L10C0QilDuo mass spectrometer (Thermo Finnigan, San
Jose, CA, USA)
PEoSs!itnieognat3ive mass-mass detection
CIoslolliastiioonn weindetrhg:y:1.350%
MESz! neg5at1iv2e0m3a8ss82de,le4ct1i9on0
PMozsition43992
Taz
NOTOX Project 334338
Conditionsfor the analytical method used in November 2002
Column CMoobliulmenptheamspeerature
FlCoolwumn temperature AInujteocstiaomnpvloelrutmee:mperature Detection
BKeetyasstilonCe1)8,50 x 2.0 mim; 6,5 ym (Thermo Hypersi, 25C
minutes) 20 mM Ammo%nium acetate
[CoTe "770
[TT 0
ss1 s|
rs
5 7" 7] :
[ CeooT Te e 7110 0 ||
23550Cymin
i10c4
SCIEX MSMS system AP1-300 mass spectrometer
Interface Montored masses
oT(ApuperprblaoitefedodnBisinpornsaeyygsatatetim4vse5,0oTno;rmonNotadof,eloCwanaatdeoaf) 7000 mimi; MMRRMM tiensftersnaulbssttaanncdaermd/mzi5z124.919.--0>-->16989..80
Standard solutions
Standard solutionsof T4127 were prepared in methanol
Calibration solutions used in July 2002
Oupn feoacmhtdwaoysotfanadnaalrydsisso,luctailoinbs.ratIinotnersnoallustitoannsdairn d50s/ol5u0ti(ov/nv)wamsetahdadneodl/tIoS2O-imnaeldicuomncweentrreatmiaodn eof 120 yg. Calibration solutions used in November 2002
On each day of analysis, calbration solutions in 15 (vv) ISO-mediumiisopropanol containing iSonltuetrinoanls.standard (internal standard concentration 164 jig) were made up from two standard
VALIDATION OF THE ANALYTICAL METHOD
w"Tahse LnoCtMcSomMpSletmeedt.hoTdhseupsaerdamweetreersonvlayipdaartteldy vaarleidgaitveedn. bUeploown.request of the sponsor, validation
T4127
NOTOX Project 334338
Validation of the analytical method used in July 2002
`Specificity sBulbasnekqIuSenOt-lmyedinijuemctwedasinpdrueptlriecaatteednatsostpheeciHfPieLdCns`ysstaemmp.leTphreetrreseualttmienngtcphrroocmeadutroeg'ra* masndwere Scroiltuitcailolny ienvtahleuastaemdefomreidntieurmfe.riInngteprefaerkisngbpyecaokmspaarriesroenquwiirtehdc{h0rboemat3og0r%amofstohfea LtOeGs.t substance
Linearity
Fmertohmatnwools/tIaSn0-dmaerddisuoml.utTihoinssr(e1s0u2l0teadnidn 1a0c6o0ncmegnt)r.ateiiognhtradnigteioonfs0.w0e50r9p--re1p.a0r0emd gin S-0/ESa0ch(voif)
these linear
solutions was injected in regression program was
urispeidcattoe.caRlceuslpaotenstehesrweegrreespsliootnteidnaegfarinosmttthheerceosnpcoenntsreastiaonnsd.
A
concentrations. The correlation coefficientisrequired o be atleast 0.99.
`Stability of the souons t`rSipollicuattei)ooofnv0es.r0a509.96,m6.g9 a,n0.d1066.1m-gh/oluranldim1e.p0e0rmigo/d,lrTe4sp1e2ct7ive' lwye.reThienjmecatxedimtuwomtdiemveisat(iionnofthe. responses was calculated for each concentration.
Validationofthe analytical method used in November 2002
Specificity
sBulabnskeq1uSe0n-tmyedinijuemctwedasinpdrueptirceaatteednatsostpheeciHfPieLdCins`ysstaammp.leTphretrreesautlmienngtcphrroocmeadutroeg'ra*masndwere. csroiltuitcailolny ienvatlheuastaedmeformeidntieurmfe.riInngteprefaerkisngbpyecaokmspaarriesroenquwiirtehdcthorboemat3og0r%amofstohfe LOleQs.t substance
Linearity
mFerdoimutmwioissotparnodapradnlsoclounttiaoinnsi(n7g8i2ntaenrndal87st4anmdaafr)d,. sTihghits driestuilotneds winearecopnrceepnairreadtiionn1r/a5n(g4eA)of1SO0pl.o0t0t5ed00a-gai0n.s1t00thmegco.nceEntarcahtioofnst.heAselisnoelaurtiroengsrewsassioinnjpercotgedraimn dwuaplsicuastee.dtRoecaslpcounlsaetse wtehere breegraetslseiaosnt 0li.n9e9from the responses and concentrations. The correlation coefficient i required fo
Blank ISO-medium (5 mi) was spiked using 50 ulof a 1.00 ma, 10.0 molor 60.1 mal solution oafntdh0e.t5e9s5t smuoblsttaesntcesuibnstmaentchea.noAlt, aylieclodinncgenctornacteinotnralteiveolns,lesviexlssaomfp0l.e0s0w9e91romgplr,ep0a.r0e9d9.1Tmhegs!e Straimplpilcaetse.weTrheeretcreoavteerdyawsadsesccarlicbueldatiendf`osramepalechprsealmrpelea.tmeTnhtepmroecaendrureec'ovaenrdya(n1a=l6ysiedtriinplicate) at each concentration level is required to be in the range 70-10%
itor tsrtl saannddwwaassaaadddrooddoda3ffoolccoonnccoonntvaatioonn 116240,s0h1
Tarr
NOTOX Project 334338
Limit ofquantitation(LOG) T{ahnegeLO7G0-w1a1s0%deatnedrmtihneecdoaatsitiheenltoowfesvatricaotnicoenntofrahtieonreactovwehriycihsthe2m0%e.an recovery is n the
mit of detection (LOD) 0.00500 mg solution of th tos substance in 1/5 (v) ISOmedium/sopropanol containing ecinaitkgeurhlntaalwteasdstafmrneodamasrtu?dhrewemdaesaasinnwjpeeecltaekads hiheniedguhpntloiaicnastede.tehveeInlmeeoafactnhhenchsoryoosmtaeetvmoe(glbroatmh,inthcepst)e.stTshuebsLtOaDncweapseak
'DATA HANDLING --VALIDATION OF THE ANALYTICAL METHOD.
Responsa:
Peak area test substance
Poak area intemal standard
Moan
7-n1it3.
where: measured valve W :number of measurements
`Standard deviation:
Gosticient of variation: Maximum deviation Uinearty
uly 2002) Validation of analytical method
> x=?
Sno =| E1-- ) (standard ovat/imoeann value) = 100% W{h(eirgehe`smt-ealnotwiestih)emmeaena]n+va1l0u0e%o the highest and the lowest value. Arorgergarsseisosnionneporfogcruarmvewafsroumstehdetroecsaplocnuslaetseatnhde Cuosnicnegntthratlieonass.sRqeugarreesssimeotnhaonda.lyAsi(s1w/caosncpeonrtroartmoend) weighting factor was used in November 2002.
Regression curve: Y = 1X + g X+ h
amt ws ad olconcarion164451.
T4127
NOTOX Project 334338
Validationofanalytical method (November 2002)
Recovery: Limit of quantification (LOQ): Limit of detection (LOD):
Regression line: Y =a X +b. wYh:ere: response aX. scopnceentration bi intercept f,.h: regression constants (1C0o0nc%entration analysed Concentration prepared) + `"Tahcecelpotwaebslte croenccoevnetrryaatinodncoofefTf4ic1i2en7t tofesvtaerdiawtiitohn.an T"T4he1l2im7itwoitfhdeatsecitginoan(ipsedaekfihneeidghats) tofhethcroenecetnitmreastitohne of noise level (SIN=3). Liofmdeteictiton= ((3 noise level) signal) conc. nwhoeirsee:level (N) : heightofthe noise [cps] csiognnca.l:(cS)o:ncheenitgrhattoifontohftteesstt ssuubbssttaannccee [pegafk][cps]
DATA HANDLING -SAMPLE ANALYSIS
Calibration
Response:
RemPeeaskmamremamtemsmt sumbsetamncees Peak area internal standard
Calibration curve: (July 2002)
Ategression program was used to calculate the croengcreenstsriaotniocnusr.veRefgrroemstshieonreasnpaolnyssiesswaansd performed using the least squares method. If necessary, a w(1alscounsceedn.tration) or (1/concentration?) weighting factor
R =asC+b R =faCiegeCeh
R: response calibration solution : concentrationoftest substance in calibration 2,0, so3l,uth:ironeg[rmegs/s)ioncoefficients
cOonnsetarcuhctdeadyusofinagnaeliygshits,coanccealnitbrraattiioonnscuinrjveectoerdliinne was. triplicate. The coefficient of correlation was > 0.99.
7 Sea DATA HANDLING -SAMPLE ANALYSIS"
Tatar Catbraton curve:
(November 2002)
Samples
Recoovfreecorveryy samples: Concentration lave fonominal; Concentration analysed: (Quadratic regression uy 2002) Concentration anaysed (Linear regression July 2002) Concentration analysed:
(November 2002)
NOTOX Project 334338
Th response was correated with the concentration of SRaua=resamasicneedd) test substance, using linear regression analysis (least R = rCoenscpeonntsreatciaointnratthoencsaotlburtaitoinon a5S= sesoipeoelnti[nmgaf] aOneedaocrhqduaayntoifficaantailoyns.isB, owhoccalablrbartaitoonnosioloutniosnswweerree iCnijesvcatmewdpal(seisnc,douUnpslsiincniagtcehi)edbfeofuorrereasnpdonafsteesr,acmaarxaimtuomnof
CCoonncceennttrraattiioonn panraelpyasreedd 100 (x) CCoonncceennttrraattiioonn nanoamliynsaeld 1004)
Cuan TE IFT ETO ER) oy
G0:BR2o2b fog
`aR nton irreegsorpeonsnssifeoancstacomorpnlsean[tunsits] CaeBR2b2 fog
aR4::: srdeiosuppteoonnnsfieacsstaoimrpnl)e [units]
be intercept units]
T4127
NOTOX Project 334338
RESULTS-VALIDATIONOFTHEANALYTICALMETHOD
"rTehsepocnalsceusl.atTihoenrseffoorret,hesvoamleidadtiifofnerteensctesswmeirgehtpebrefoorbmseedrvuesdinwghneont-craolcuunldaetdincgotnhceesntattirsatitcialaonsd parameters using the values as mentioned in the tables.
Validation of the analytical method in July 2002
`Specificity cFoingtuariensin1gainndte2rnsalhostwacndharrodm)aatnodgroafmas1o.f0a0 bmlgankTs-o4l1u2ti7osno(luSt0i/o5n0,(rveAs)pemcteitvhelayn.oIltIwSaOs-mcleedairutmhat blank chromatograms did not contain any interfering peaks at the position of the test substance.
Linearity The results are summarized in Table 1. The regression fine is shown in Figure 3.
Table 1 Linearity.
Concentration mo)
Response units]
00.00751094 0.0120064 0.0460132 01.08006
0000110491//00.00114092//00..00104888 00..00420185//00..00240077//00..00242446 0.00.7008179//00..01702368//00..10069370
00..112465//00..115330//00..115168
+ Trploata measurements. 1`0T.h9e9s5e6)rebseulttwseesnhorwesthpaotnstheearendiscaonrceelnattriaotnsihoinpi(nYth=e0.co0n0c2e4nt4r+at0i.o1n84r6anXge- o0f.00.3075198 X'1;.R00' m=g/ `tohbosuogrhveaddaetvivaatriioonusofcomnocernetrtahtainon1s.0% of the calibration points from the calculated line was.
T4127
Stabiltyofthesolutions
The results are summarised in Tables 24.
Table 2 Stabiltyof 20.0509mg/l T4127 solution.
Eapsed ime Tours]
Response
0020
0000110029
9015
0000100378
9964
000001305s
NOTOX Project 334338
Wiasimum deviation tl 142
Table 3 Stability of 20.106 mg! T4127 solution
Eapsed ime hours]
Response
0020
0000221057
8045
0000221276
8879
0000220248
Wiasimum deviation tl 10
Tabl4e Stabiloftay 1.00 mg/l T4127 solution.
Eiapsed me hours]
Response
0002
00.11543508
5058
00..11558539
6519
001147238
Maximum deviation [13]
9T6h-ehsoeurresiumltesisnhteorwvatlhaat athceonscoelnuttiroantsiowneorfe0s.t0ab5l0e9(mmalxiamtulmeadsetvaia8t.i0on-h<o2ur0%m)eovienrtearvalleaatsaat concentrationof 0.106mg and atleasat6.1-nour ime interval ata concentration of 1.00 mol.
T-4127
NOTOX Project 334338
Validation of the analytical method in November 2002
`Soecticty cFoingluariensin4gainndte5maslhsotwancdharrodm)aatnodgroafmas0o.f5a00blmagn!k Tso-l4u1ti2o7ns(ol1u5ti(ovnv,)re1sSpOe-ctmievdeilyu.m/iswoapsrocpaanrolthat blank chromatograms did not contain any interfering peaks atthe position ofth test substance.
nearty The results are summarized in Table 5. The regression ne is shown in Figure 6.
Tables Linearity.
Concmeroinraton
Resupnoins]se
000000659090 0000119090 000.000784800200 0.100
000.714021//000.8074247 00217227//00216206, 000587698310///000598550823 123/116
IntercepSttwoipeh Y-axis `WeightiAng factor
01021134 Vcon0c9e9nr7a9tion
+ OOuupric:otsomtosasaudron mans. cfha cabria o cue cFornocmentthreastieornesiunlttsh,e tcwonacsenctornatcilounderdantghaetotf h0r.e00s50a0n-e0a.r10r0elmatgi!onsh3ip(bceontcweenetnrarteisopno)nsweeiagnhdting factors used.
T4127
NOTOX Project 334338
Accuracy and Precision - bil
1SO-medium spiked with T-4127 to concentrations of 0.00991 mg, 0.0991 and 0.595 maf (and 164 gl internal standard), was pretreated as described in `sample pretreatment procedure' Chromatogramsofpretreated 0.00991 mf, 0.0991 and 0.595 mg/l samples are shown in Figures 7, 8:and 9. The resuls are summarised in Table 6.
`Table 6 Accuracy and Precision.
Daleci [Date | Conc. | Conc. Recovery' | Mean | Cosficient
Preparation| Analysis | prepared | analysed [%) | Recovery| of variation
(d-mm-yy) | (dd-mm-yy) | (mo) [mg]
%]
%]
.
15-Nov-02| 15-Nov-02| 0.00991 | <LOD
na.
na
na.
<LLooDp nnaa..
<<LLooDp nnaa..
<Lop na
15Nov-02| 15-Nov02| 0.0891 00.111114
111152
1 24
0118
119
on7
118
0118
119
0.118
118
15Nov-02| 15:Nov-02| 0595| 06442 108 1"
18
00g6648172 11028
0066780877 1113
o0s627 hid
TCroolcalteembeyuasxuarteamtpeonaettso.dn.MRaeismpuonnsdeesvwiaotiroen<o1f1p0l%aotfahmeeNaisguhrceambernattsownasev<el20%
c`Tohnosuigdhermeedaanccreepctoavbelreiefsorwsearmepallles(silnigIhtSlOy-)mheidgihuerrnthinanth1e1c0o%n,cetnhteraantailoyntircaanlgmeeotfh0o.d09w9as1.- 0.595 mg! because the coefficient of variation was <<20%.
Limit ofquantification (LOQ)
T1h7e%,LOG for the samples in ISO-medium is reported to be 0.0991 mg though recovery was.
T4127
NOTOX Project 334338
Limit of detection (LOD) (FNr)owmatswodectherrommianteodgtroabmes 2of225 0c.ps0.05F0r0ommoltlhesoslaumtieonchofrothmattoegstrasmusb,sttahnecet,esthseubmsetaanncneoissiegnlaelvel (t5h)e.sie.vatlhueesm,etahne lhiemiitghotfodfettehcetiteosnt(sSuNb=s3ta)nwcaespecaakl,ciwaatseddettoebremi0n.e0d03(09 bmeg!87a.l5acnpsi.njUescitniogn vthoelulimmeitooff1d0etuel.ctBioencafourstehsesaempslaemsplfeosmisth0e.0e2c3otmoxgi.calogial study were died by a factor of 5,
RESULTS - SAMPLE ANALYSIS
Samples July 2002
Table 7 shows the analytical results for the samples from the test performed in July 2002"
Table 7 ConcentraoftTi-o41n2s7 in test medium in July 2002 (Ful test 1)
sTaimmpleionfg|| sDaamlpelionfg | aDnaatleysoifs [~Norinal hours] | fecmm-yy]| (ekmm-yy]| [mo]
Analy]sed
nRoemlinaatle[T6o]
0 | otwez | o2uu02* 1000?
n1a.0.30
n1a.0.30
100
0447
as?
20 | 0202| o2Juk02 1000%
0nd5.62
0na5.62
100
0178
0.178
% | 0502| O5Juko2 1000?
0n3d32
0na3.32
100
002
0082
"Mon Sm ofredlesaotoannsyespa.rTahamaaaniomnedevciaotniocnnbowenft1h00reoslponses calaaedfor sac samplewas < 20% 2 SaCmoplceesdwobryeapxsveaaettetdono.n R0e1s3p0o-n0se2s3wdohra2d8to05%a.fsohreodwcevse:mcihlt 1antgeever.sor gehts h calbrsion
SSoluttoerrs. cue snailprobly lmsandrwsa4 1 esssample nd caibrason
Ahloan tcvovllutosthwereiceouiutsu0si5ng0othroocuonndceednrcaotonnscae3nm.onThereie,hasaaem.deans be esa
T4127
NOTOX Project 334338
Samples November 2002
Table 8 shows the analytical results for the samples from the test performed in November 2002. Table 8 Concentrations of T4127 in test medium in November 2002 (final test).
sTaimmpleionfg|| Dsaamtpeloinfg | aDnaatleysoifs hours]| [de-mmoyy]| fod-mm-yy]
`Analysed Img
Reiatve to nominal (%]
07 | 11-Nov-02| 11Nov02| 110000 07 | 11-Nov-02 | 11-Nov-02 00
00.003344 00007766 00.1155 003344 007766 48 | 13-Nov-02| 13-Nov-02 00 0.034 00.007364 000.1756 00.3145 003746 078
00777500
nnad.
nnaa..
00.0022142%
661
00.002694
838
00.1136
18073
002240
5790
007556
874
nngg..
nnaa
<<LLoopp
-5
00002213
227
00.003484
22
00.007833
221
003310
329
T4127
NOTOX Project 334338
(Table 8 Continued) Tsiammpelionfg|| Dsaatmpeionfg | Danaatleyosifs [Nominal hours) | [de-mm-yy]| [dd-mmyy]| (moll
"Analysed [mg/l
Relaivelo nominal (%]
15-Nov-02
00
nndg..
nnaa..
00..003344
<<LLoopD
::
00.007766
<<LLOoDD,
B:s
00..1155
0000211973
113
003344
00.004409*%
124
Muenalenssofatihpoircwaitsenndlaytd..Themaximumdeviationbebweanth responsescacuatd oroachsamplewas <20% 2> CComabilnedcbwuyiehxaNtaOptTesOoaXtpordno.jeRcet3sp4o3n4se5swerabetween50and 100%of he lowestcalbraton ove.These
concaniationswerno reprtdas <LODbecausethey wers anysightbelow hemi ofdetection. Maximumdeiatonbetween responseswas 23%,36%.32%,22% and22%,respecivey. nnad.. NNoottdaoptpoicceadb.eThe itofdeectn (LOD)was0.023ml.
Tar
NOTOX Project334338
j ol Samples July 2002
ar 0001230 oy
= Internal standard
he |
fo3l
fal
|
sroT0R
Scomoaneune
334338232
:
n A03m L ---- ee -- eee
o=m3 Testsubstance
Bene
I
T[riRossasontn
iv: 03
3
lhl|
pa Figure 1 HPLC chromatogram ofa bank (50/50 (v) methanolISO-medium containing
internal standard).
T4127
aroowcedo3. 1330
of
| 70] Internal standard
I|
3
il
NOTOXProject34338 wE noernsn ern o SSEsh
;
u
| Testsubaance
|1|
pjrHreyaRseD cen
ahfrasrte i
fioel
peol
OTT
TTEaTtTo)
Ii
JISe
TTT TT
Figure2 HcoPnLtaCincihnrgoimnatetrongalrasmtaonfda1r.d0.0 mg/l T-4127 in 50/50 (v/v) methanol/ISO-medium
T4127
NOTOX Project 334338
Y=0.00244219+0.184632X0.0T3471127766X'2 RA2=0.9959 W: Equal 0165
0147
i
0125
20
g 0.08
.
< 006
re
oo
"ro
0.0: Ie
00038 -------- 00 02
04
: 0s
08 rr 10
mon
Figure 3 IRnetgerrneaslsistoannidanrodf:orResoslpuotinosnessinag5a0i5ns0t(cvovn)cemntertahtainonosl.1SO-medium containing
Taz `Samples November 2002
EBEtr i | aie
NOTOXProject334338
B[ErE]|
en
faa
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Fre Ho PLC van mmatogag monofda0691mgT4127in U5 (1) ISOmadumisopropond
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