Document Yj30LNoL3VrjjQd6Qg6KKOd5V
AR226 -- 3256
438
AR4RE - 3256
DuPont-10819
TRADE SECRET
Study Tite
"NR5:cteric!Reverse MutationTestwithanIndependent Repeat Assay
Authors Val`eSnatmiuneelOA.tWtaa-gSnaefro,h,IBL',SM..S.
Study CompletDiaotne 16 September 2002
PerformingLaboratory
9630 MeBdiiocRaelliCaenncteer Drive.
Rockville,foMr D 20850
E.L duDPuoPnotndteHaNsekmelolurLsabaonrdatCoormypany
-
109P0.OE.lkBtooxn 5R0oad
Newark, DE 19714-0050
"PerformingLaboratory StudyNumber
AAGOES502001 BTL
WorkRequestNumber
Servies Code:
~
Page Tof65
--
Company Sanitzed. Does not contain TSCA CBI
TE stoin
estwithan Independent Repeat Assay
DuPont-10819
STATEMEONFT COMPLIANCE
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BioRelance Study Management
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BioRelizos EE
Company Sanitized Doss not conan TSCACEI
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Q5E::cric] Reverse Mutation
`Test with an Independent Repeat Assay
'DuPont-10819
Quality Assurance Statement Study Te: BAAsCsTaEyRIALREVERSE MUTATION ASSAY WITH AN INDEPENDENTREPEAT ASSAY
stay numer Study Director: ValainOe. Wagaer, I, MS
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DuPont-10819
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Thsorreporsdeyesthe motosandprocedureswd nthesud and he ered slscurerfl hee
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BioRel Study No.
4
Company Seniized, Doss ot contin T5GA Go
--~ cin
WHEE Independent Repeat Assay
Dupont 10815
CERTIFICATION
`We, the undersigned, declare that this report provides an accurate evaluationofdata
`obtained from this study.
sued by Study Director:
Natetze OU. ser, Ti Sep acon
`Valentine O. Wagner ITI, M.S.
Date
StuAdpypMroonvietdobry bee Dore
06 sePrmL
SenioMrarRieaseDaorncnherT,orPiho.Dl.ogist
Date
--
BRE.
--
study No. SE 5
ee ee e AA e re Lee
~~
cov usin
est wit IndependentRepeatAssay
`TABLE OF CONTENTS
DuPont-10819 Page
CharacterizationofTest and CORIO! SUBSANCES...vvvvcerssrrssssersomssssssessnssssssssssssssns 10
~~
--_--
"Table 1: Preliminary Toxicity Test in Salmonella typhimurium TAS ......o.ceevuvcnrs 19
"Table 2:
Table 3:
Preliminary
Preliminary
Toxicity
Toxicity
Test
Test
in
in
Salmonella
Salmonella
typhimurium
typhimurium
TA100
TA1535
.......oc..cwcrne..
o.oo
20
21
`Table 6: Mutagenicity Test in Salmonella typhimurium TA98 Without $9 .......c........ 24
Table 7: Mutagenicity Test in Salmonella typhimurium TAS With S9 ..........o.r.25
Table 8: Mutagenicity Test in Salmonella typhimurium TA100 without S9 ...............26
Table 9: Mutagenicity Test in Salmonella typhimurium TA100 With S9 .........cco..... 27
`Table 10: Mutagenicity Test in Salmonella typhimurium TA1535 without 9 ............ 28
`Table 11: Mutagenicity Test in Salmonella typhimurium TA1535 With S9 ..............29
`Table 12: Mutagenicity Test in Salmonella typhimurium TA1537 Without 9 ............ 30
`Table13: Mutagenicity Test in Salmonella typhimurium TA1537 With S9 ..c...oer 31
`Table 14: MutagenicTietsyt in Escherichia coli WP2 uvrA Without $9... 32
`Table15: Mutagenicity Test in Escherichia coli WP2 uvrA With 89.......c..covvurrnnn 33
"Table 16: Mutagenicity Test in Salmonella typhimurium TA98 without S9 ................. 34
`Table
`Table
17:
18:
Mutagenicity
Mutagenicity
Test
Test
in
in
Salmonella
Salmonella
typhimurium
typhimurium
TA98 With 89 ............coo..
TA100 without $9 ............
35
36
`Table 19: Mutagenicity Test in Salmonella typhimurium TA100 With S9 v......ver. 37
`Table 20: Mutagenicity Test in Salmonella typhimurium TA1535 without S9..............38
`Table 21: Mutagenicity Test in Salmonella typhimurium TA1535 with S9 ................. 39
`Table
`Table
22:
23:
Mutagenicity
Mutagenicity
Test
Test
in
in
Salmonella
Salmonella
typhimurium
typhimurium
TA1537
TA1537
without
With 9
S9 ............. 40
......c.......41
sway NoJE BioRelianc
cCoompmanmSeates. cpeecsotmecwnseamsoocan
I 5c. Rovere Mutation TestwithanIndependentRepeatAssay
DuPont-10819
`Table24: Mutagenicity Test in Escherichia coli WP2 uvrA Without $9........cuuvunn 42 `Table 25: Mutagenicity Test in Escherichia coli WP2 uvrA With 89......cc.ccuwssssnnn 43
`Table26:Salmonella/E.coli Mutagenicity Test -- Summary of Results Bl .............44 Table 27: Salmonella/E. coli Mutagenicity Test -- Summary of Results B2.............45
Appendix B: SAY POOCOL...vcvvvivsrmssmssssssssssisssssssssssssssssssssssssssssssssssssssnss 48.
Appendix C: Informationfor Japanese Regulatory AGERCES.........uwwswessescs 60
PR
sBiuodRyelNioa.ncEe E
7 Rm a a
--
esIt witDh an Incdepcendenet RrepseaMt sAsssiaoyn
DuPont-10819
STUDY INFORMATION
Substance Testes:BEN
Haskell Number; 25339
`KnownImpurities: {IF
Physics] Characteristics: Yellow dispersion in water
~~
`Stability Tchonedtietsiosnusbosftatnhceesatpupdye;arneodetviodbeenscaebolfeuinnsdtaebritlihtyewas
observed.
-
`Sponsor: EW.iLlmdiungPtoonnt,dDeeNleawmaoruers19a8n9d8Company
USA.
`ISntuidtyiated/Completeds June13, 2002 (seereportcoverpage)
InI-nLiitfieated/Completed: June 18,20/ J0ul2y 19,2002
BoRebmes Study No.
5 Sompany Saritized, Dogs ool conlain TSCA CE
~~
TE estwithanIn5:dceipcerniR dclenRteRveepresaetMuAtsastaiyon
DuPont-10819
SUMMARY
"Thetestsubstance,H-25339,wastestedinthe BacterialReverseMutationTestwith an ITnAdLeSp3e5ndaenndt TRAeLpSea3t7aAnsdsEasychuesriincghiSaaclomlonteelstleasttryapihnimWuEr?iuumrtesitnetrhestprraeinssenTcAeSaSn,dabTsAeLnOcDo, ofAroclor-inducedrat liver $9.The testwasperformedin twophases, usingtheplaie incorporation method.Thefirstphas,thepreliminarytoxicity est, wasusedtoestablishthe doserangeforthemutagenicity test.Thesecondphase,themutagenicityfest (inital and independentrepea tests)wasusdtoevaluatethemutagenicpoteaialofth testsubstance.
`Waterwasselectedasthesolvent ofchoicebasedonthe Sponsor's request.
wasIancthhieepvreedliumsiinnagryatocxoinccietyntterasttitohnoe mfaxS0immgu/mmdLo.saentedstaed1w0a0su5L0p0l0atiignpgearlpilqautoet;.tThhsedtoesset substancewassolublebutcloudyinwater atapproximately50 mg/mL. The doselevelstested were 67, 10,33, 67, 100, 333, 667, 1000, 3333 and5000 ug per plate. Neitherprecipitatenor daopsperepcliaatbeldeintotxhiecmiutytawgaesnoibcisteyrvteedt.wBaassSe0d0o0ntuhgpefeinpdliatneg.softhe (oxicity est, the maximum
--
Inthemutagenicitytest, nopositive mutagenicresponsewas observed. The doselevels
tested were 100, 333, 1000, 3333 and 5000 pug per plate.Neitherprecipitate nor appreciable
toxwiascobsiertveyd.
"Theresuls oftheBacterial Reverse Mutation Testwithan IndependentRepeatAssay ienidtihcearttheethparte,suenndceerotrhaebcsoenndcietoifonAsorfothcislstourdy-,iHr-na2td5l3iu3vce9rdei9dd.notcause positive fesponsein
BioRelianee
soyNoUN Company Senilzed, Dosa coniainTSCAGE
~~
SE 5: Reverse Mutation
"Test with an IndependentRepeat Assay
DuPont-10819
PURPOSE
`Thepurposeofthisstudywastoevaluate the mutagenicpotentialofthe test substanbcye
mSeaalsmuornienlglaittsypahbiimlutryiutmoainnddautcethreetvreyrpsteopmhuatnaltoiocnussaotf EsseclheecrtiecdhilaoccioloifssevterarWl Psa2trwariinAs noifn
thepresenceandabsence AppendiBx.
ofAroclor-inducedrat
liver
9.
A
copyof
theprotocolisincludedin
TBahcetersitauldyRevwearssecMountdautcitoend Tteos)c,oamdpolpyiedwiJtuhlyO1E9C97D(pGuubildieslhiendeFe4b7r1uar(yGe1n9e9t8i)c aTnodxiwciotlhogtyh:e IPntehmataionral mCofnafoerrcHeunmceeanounUstHea(ri1m9o9nci6saaantdi1oln99o7sf).Technical Requirements of Registration of
CHARACTERIZATIONOFTESTANDCONTROL SUBSTANCES
Thetestsubstance, H-25339, wasreceivedby BioRelianceon 12 June2002andwasassigned
S diseperlsionI in w dbest2 oreds atamwbyiec nttems perati ure. AonneSxpinratrionsdpawteoowf
--_
0ye1lOlcowtcolboerud2y0l0i2quwidaasnpdrwoavisdsetdo.reUdpaotnoroecmeitpetm,ptehreattuersetasnudbisntatnhcedeawraks.describedas apale
`hTahveebideeentnidtye,tsetrremnigntehd,bpuyrtihtye,cSopmopnossoirt.iTohneosrtaobitlhietrycohfartahcettereissttsiucbssttoadnecfeihnaesthbeeetnedstestuebrsmtiannecde. bythe Sponsorbut acopyoftheanalysiswasnotprovided to BioReliance.
`7T7h3e2v-e1h8i5c)l,e uosbetdatinoeddeflriovemrTHa-vi2t5r3o3g9entoCtohrepotreasttisoyn.steTmeswtassusbtsetrainlcedeisdtiillulteidownsatweerre(CpArSepaNroe.d immediatelybeforeuseanddelivered1 thetest systematroomtemperatureunderyellow light.
`iTlheewniegtahttihveeatenstdipnogfsatciivlietcyo.nTtrhoel ssutabbsitlaitnycoesfhtahveneebgeaetnicvheaarnadctpeorsiizteidvaescopnetrrtohlesCuebrsttifainccaetseasonnd theirmixtureswasdemonstratedbyacceptableresultsthatmeetthecriteriafor avalidtest.
Potsabiltei.vAelclonptorsoiltispvelactoendtcroolnscuwrerreentdliylwuietdhtwihtehmudtiamegtehnyilcistuyltfeosxtiadere(lDisMtSeOdi)netxhceefptolsloodwiiunmg aazti-de5,10w-h3i0chCw.asdiluted withwater,Allsubdivided solutionsofpositivecontrolwerestored
sBiooReoliance
ee
SompenySanitized, Dogs dolcouain TSCACE
a]---- [m --
EE: 5c! Reverse Mutation
Testwithan IndependentRepeatAssay
5
DuPont-10819 Concentration
Al Salmonella Strains.
(Aldr2i-cahmCihneomaintcharlaCceon,eInc)
We2urA
EspL.oDtaNtoe.1049-1S0ep6-P2S005
CAPSuriNtyo. 961734-%138
(Adsic2hnCithreomfiicuaolreCnoe,Inc)
TA9
LotNo. 08707HS
Exp. Date 08-Mar-2006
`CAS No. 607-57-8
10
Purity 999%
(SigSmoadCihuemmiacziadleCo)
Taunzams
EsLpo.tDNaot.e 0059-8JHa0n1-620904
'--_
`CASPuNrioty. 29696.258%228
TALS
[Fee2iozs
(SLiotgmNao.Ch1e0m6iFc0a6l6C8o1)
ExpC.ADaStNoo1.85-0N4ov5-.29004
=
Pusity 507%
Methyl methanesulfonate
(AldrLiochtCNhoe.m1ic5a5l2C6oA.0 Inc)
ExpC.ADSatNeo2.66-6N:o2v7--23003
Purity 99.9%
themTuotacgoennfiircmitythtesttewrailsiptyloaftetdhoetneseltescutbisvteaangcaer,twhiethhiganhaelsitqtueosttsuvbosltuamneceqeudaosefoletvhealtwuseeddiinn the est.Theseplatswer incubated underthesameconditionsasthe test.
sors
Company Seritzed, Dogs nol coniaia TSGAG
- SI cic Reverse Mutation "Test with an Independent Repeat Assay
DuPont-10819
MATER ANI D MA ETL HOS DS
Forsubmission to Japanese regulatory agencies, additional information is included in AppendiCx.
`TestSystem
`ThetesterstrainsusedweretheSalmonellatyphimuriumhistidineauxotrophs TA98,
TAI00, TA1535 and TA1537 as described by Ames ef al. (1975) and Escherichia coli 'WP2uvrAas describedbyGreen and Muriel (1976). Salmonellatester strainswerereceived directly fromDr. Bruce Ames,UniversityofCalifornia, Berkeley. E. colitesterstrains were
`receivedfromthe NationalCollectionofIndustrialandMarineBacteria,Aberdeen, Scotland.
Tester strains TA98and TA1537 are reverted from histidine dependence (auxotrophy) to histidine independence(prototrophy) by frameshiftmutagens. Testerstrain TA1535 isreverted bymatagens thatcausebasepairsubstitutions. Tester strain TA100 is revertedby mutagens that cause both frameshift andbasepair substitution mutations.Specificity of thereversion
`mechanisminE. coliis sensitivetobase-pairsubstitution mutations,ratherthanframeshift `mu(Gt reea n at ndMi urio el,n 197s 6).
Overnightcultureswere preparedby inoculating from the appropriate master plate or from
~~
the appropriatefrozen permanentstock intoa vessel containing ~50 mLof culture medium. To
`assurethatcultureswereharvestedinlatelogphase, thelength ofincubationwascontrolled
2motioned.Fllowiug oeaation, each Hk was Pico i a resi shefonbetor `roomtemperature.Theshaker/incubatorwas programmedtobegin shaking atapproximately
1251pm at 3742C approximately12hours before the anticipated timeofharvest. Each culture was monitoredspectrophotometrically for turbidity andwasharvested at a percent transmittance yielding atiter of approximately 10 cells permilliliter.Theactual titers were
Gcemmined by vil out esonnuient geeplate sdthe data 5 onfle. bk nt `presinethinsrtepeordt.
MetabolicActivation System
Aroclor 1254-inducedratliver S9wasused asthemetabolic activation system.The S9
:
`waspreparedfrommale Sprague-Dawleyratsinducedwith asingleintraperitonealinjectionof
Aroclor1254, 500mg/kg,five daysprior to sacrifice.The S9wasbatch preparedby
BioRelianceorpurchasedfromMoltoxandstoredat -70Corcolderuntilused.Eachbulk
prdepi aram tioe n ot f Sh 9wy asltesb ttoefe dofron mrsimz tustaa( bgielna iitcy) ttooSa maeltmnaobnet olllih aztey2r p-haimamiunrc oiaune mtThrAna1c0ee 0n.e and 7,12-
`TheS9mix waspreparedimmediatelybeforeitsuseandcontained 10% S9 5m,M glucose-6-phosphate,4 mMB-nicotidnina uclm eotiidedphe osp- hatae,d 8meMn Mgi Cln ;ae nd
BsiooRierlNioanG ee__E __--E-- ,
Pr
------
~~
I: ci ReverseMutation
Test withan IndependentRepeatAssay
DuPont-10819
33mMKClin a 100mMphosphatebufferat pH 7.4. The Sham S9mixture (Shammix),
`containing 100mM phosphatebufferat pH 7.4,waspreparedimmediatelybeforeitsuse.To
`confirmthesterility ofthe S9and Sham mixes, a0.5mLaliquotofeach wasplatedon selective agar.
SolubilityTest `Waterwasselectedasthesolventofchoicebasedonthe Sponsor's requestand
`compawitthithbe tiarlgeitcetllys.
Preliminary ToxicityTest
Thepreliminarytoxicitytest wasusedtoestablishthe dose-rangeoverwhichthe test
`substancewouldbetested.Vehicleandtendose levelsofthetestsubstancewereplated,one
Y
`plate perdose, withovernightcultures of TA98, TA100, TA1535, TA1537and WP2wvrA on
selective minimalagarinthe presence andabsence of Aroclor-inducedratliver 9.Thedose
levelstestedwere6.7, 10, 33, 67, 100, 333, 667, 1000, 3333and5000 pgperplate.
`Mutagenicity Test
--
Fiv`eTdhoesmeulteavgeelnsicoifttyettessutbwsatsaunsceedaltoonegvwaliutahteatphperompurtaaigveenhiiccploteeanntidaploosfitthievetceosnttsruoblsstawnecree.
plated with TA98, TA100, TA1535, TA1537 and WP2 wwrA in the presence and absence of
iplcate. Aroclor-indurcaetd liver S9. The dose levels tested were 100, 333, 1000, 3333 and 5000 pg
`perplate. Alldose levels oftest substance, vehicle controls and positive controlswereplated in
PlatingandScoringProcedures
`Thetestsystemwasexposed tothetestsubstanceviatheplateincorporationmethodology
originallydescribed byAmesetal. (1975)andupdatedbyMaronandAmes(1983).
On thedayof itsuse, minimal topagar, containing 0.8 %agar(W/V)and 0.5 % NaCl
(W/V), was melted and supplemented with L-histidine, D-biotinandL-tryptophan solution to
saufpipnlaelmceonntceednwtiratthi2o5n moLfo5f0wuatMeerfaocrhe.acThop1a00gmaLrnooftmuinsiemdawlittophagSa9ro.FrorSthhaemprmepiaxrawtiaosn
`ofmedia andreagents,allreferencesto waterimplysterile,deionizedwaterproducedbythe
Milli-QReaWg atee rSyn stet m.Bottomagarwas Vogel-Bonnerminimalmedium E(Vogel
`amnidniBmoanlnmeerd,iu19m56E)cocnotntaaiinniinngg1.15.%%5(W(/WV/V))agaagrara.ndNusturpipelnetmbeonttteodmwiatgahr2.w5as%(VWog/eVl)-OBoxnoniedr
-
Nutrient Broth No. 2 (dry powder). Nutrient Broth was Vogel-Bonner salt solution
`supplemented with 2.5 % (W/V)Oxoid Nutrient BrothNo. 2 (dry powder).
soBiioRyelNioan.ce.EE
CompanySoriied, Gosia contain SAGE
~
I ccoVion
est with an Independent Repeat Assay
`DuPont-10819
doseEalecvhelp,latteeswtearssltraabienl,eadnwdiatchtiavcaotidoens,yasstedmestchraitbieddenitnifdieedtatlheitneBsitsouRbesltiaancnec,et'esSttpahnadsaer,d OperatingProcedares
ortOensets-uhbasltfan0c.e5d)imliultliilointweerorfea$d9doerdSthoam2.m0imxL,o1f00moklLteonfsteelsetcetrsitvertaionpaangadra1t004k5L:o2fCv.eAbfitcelre vportlexaitnght,etphioesmniitxigvteurcewaosonvettrhlearitdesootnstulobtshsteasn,ucrefaacleioqufot25wmasLrofemipnilmbaaylbacotSetOomdLaaglairq.uWohtoefn appropriate positivecontrol.Aftertheoverlayhadsolidified,theplateswereinvertedand imimnecdubiaatteedlfyofrolalpopwrionxgitmhaetienlcyu4ba8titonop7e2rihooduwresraetst3or7e4d2atC2.-P8laCtuensttihlcaotlwoenryceonuontticnogucnotueldd becond(ieusstchant10edayds).
Thecondition of the bacterialbackground lawn was evaflorueviadentce oef tdest substance toxicitybyusing adissectingmicroscope.Precipiatewasevaluatedbyvisual examinationwithoutmagnification. Toxicity anddegreeofprecipitationwerescoredrelative. tothevehiclecontrol lateusingthecodesshowninthefolowingtable.
-- [TTNSoigmhatyl [Di_st|inDguEisihe2dbbgyyua2hineoastlitchheyaembildecrotcohloniyolfanwthne.nm_icir_ocno_ logay_Ta|w
Regucey |comdparedposstiobtlahyevselbiigchltecinocrneatseprliaonttelh. esizeofthe microcolonies|
I"
Distinguished by a marked thinning of the mictocolony Tawa
3
[resultingin a pronounced increaseinthesizeofthe microcolonies|
[Distinguished by an extreme thinning of the microcolony lawn] 4 | Extremely|rsulinginanincreaseinth sizeofthemicrocolonicscompared
Reduced tothevehiclecontrolplatesuchthatthemicrocolony lawn is vitsotihe ubnaildeedeye as isolated colonies.
[aEr greater thanorequal to90% ofthe plate. [asy Nop. |itingaished by precipita on the plate that is Visble to the
Inrioing [22k ye butanyprecipitateparticlesdetectedbythautomated Presgiug colonycoutontaltesethran 10%oftherevertant colony coat
e.2. lessthan 3particleson aplatewith30revertants).
SBiuodRyeNlioa.nceEE
+
Company Sanitized. Does not contain TSCA CE)
~
CP ccc Revs sain
"TestwithanIndependentRepeatAssay
DuPont-10819
[Code T Description
Characteristics _______]
nering |P[CaDlkiosentdiyencgyoueuisnahtneeddrabnyeyxpcprreeeeccdiippi1itt0aa%tteepooafrnttithchlreeospvdleeatttaeenct(thceadotbliyosntvhyiesciaobuluteonmtoa(t0te5hde.||
PI |morethan 3particleson aplatewith 30 revertants).Theseplates
arecountedmanually.
Revertantcoloniesfor agiventester strainandactivationcondition,exceptforpositive control,werecountedeitherentirelybyautomatedcolonycountero entirelybyhandunless theplateexhibitedtoxicity.
`EvaluoafRteisuoltns
Foreachreplicateplating, themeanandstandarddeviationofthenumberofrevertantsper platewerecalcaundlaraeretpoertedd.
Forthetestsubstancetobeevaluatedpositive, itmustcause adose-relatedincreaseinthe.
`meanrevertantsperplateofat leastonetester trainover aminimum oftwoincreasing
p`ocsointcievnetriafttihoenisoncfrteeasstesiunbmsteaancner.eDvearttaasntestsaftotrhteepsteearksotrfatihnes dToAsLerSe3sSpaonnsdeTiAe1qSu3a7l woeorregjruedagteedr
-
than 3.0-mesthemeanvehiclecontrolvalue.Datasetsfortesterstrains TAS, TAI00and
WrPes2puonvsreiAweqeurael joudogregdrepaotseirttihvaein2f.t0h-etiinmcersteahseemienamneveahnicrelvecerotnatnrtoslavatltuhee.peakofthedose.
CriftoraeVarliidTeast
so SE oo 5 `Thefollowingcriteriamustbemet forthemutagenicitytesttobeconsideredvalid. All
S(aflam)oanneldltahteesdteelresttiroaniinncutlhteuresvmrBugsetndee.mCounlstturraetseotfhetpersetesresntcreaoifnsthTeAdSeSeapnrdouTgAhIm0u0tmatuisont `demonstrathee. presenceofthe pKMI01 plasmid Refactor. All WP2 vr cultures must d`emmeoannsnturmabteerthoefdseploenttiaonnieonutshreevuertAantgseinnet.hAelvlechuilctluerceosnmturoslstadesmfoonlsliorwast(eitnhcelucshiavrea)c:teTriAstOic, 10-50;TAI00, 80 -240;TAIS35, 5 -45; TAIS37,3 -21;WP2uvrA, 10 -60. Toensutrhaet `appropriate numbersofbacteriaareplated,testerstrainculture titersmustbegreaterthanof i`necqrueaaltseoi0n3txh1e0"ncuemlblesr/omfL.reTvheertmanetasnovoefretahcehmpeosaintivvaeclouenotrfotlhmeursetspeexchtiibvietvaethilcelasetcoan3t.r0o-lf.oAld mcionnsiimdeurmeodtfoxtihcriefeonnoen-otrobxoitchdoofstehleefvoelllsoiwsirnegqcuriitreerditaaoreevmaeltu:at(e1t)eAstd>a5t0a9%.rAedduocstieolneivneltihse r`emdeuacntionnummbuesrtobferaeccvomeparntipeeadrnbplytaatsenaasbcrupotdomsep-d(ea0petrnhdeeemnetdadrnovpeihnitclheecreovnetrrtoalntvcaloueu.t.T(h2i)s Atleast amoderatereduction inthebackgroundlawn(backgroundlawn code3, 4or5). A `copyoftheHistoricalNegativeandPositiveControl Valuesisincludedin AppendixA. BioReliancs _
Company Sanitized. Doss not conta) TSCA CE
~
cov vin
with anIndependentRepeatAssay
DuPont-10819
Archives
Allrawdata, the
andallreportswillbemaintainedaccordingto Standard
Opcuing Procedure the BioRelace, RAQA. unit headend. bBeiroeRtealiinaendcfeo,ra14t9l2ea0sBtrtohsrceheayretaRrosaafdt,eRrowchkivcihltlie,meMtDhe2S0p8o5n0s.oPrewritlhibsecSoOnPt,acptaepdefrorreacdeocirwsdiiolsnl
as tothefinal disposition ofthematerials. AllstudymaterialsreturnedtotheSponsor or destroyedwillfirstbecopied andthe copy willberetainedinthe BioReliancearchivesfor a
ism of 10yes. Unused dosingsotonswredisposedof lowingsaminirionfo thetestsystemandallresidualtestsubstancewillbedisposedof followingfinalization ofthe
report.
Deviations
oftissud. Noknowndeviationsfromtheprotocolortest-method SOPsoccurredduringtheconduct
suQUiyNo
Company Saritzes. boos no conta Toca cp
~~
E est iR thanIndepceindaenRtReveepresaetMAissastaiyon
DuPont-10819
RESANUD DILSCUTSSISON
Solubility Test
`WaterwasselectedasthesolventofchoicebasedontheSponsor' request.
:
Preliminary ToxicityTest
wer"eTgheenreerasttesdoifntEhxpperreilmimeinntaArLy tIoxnictihtey ptreesltaimrienparreysteonxtiecdiitnytTeasbtl,etshe1mtharxoiumguhm5.dTohseesteedsattead w1a0s05uL0p0l0atuignpgealripqluaotte.;"tThhiestdeosstseubwsatsaanccheiweavsesdoulsuibnlgebuatcocnlcoeundtyriatniwoantoefr a5t0 mapgp/rmoxLimaantdelya p5e0rmpgl/amtL.N.eTihtehedropsreecliepvietlateenstoerdawpeprreec6i.a7,ble10t,o3x3i,ci6t7y,wa10s0o,b3s3e3r,v6e6d7.,B1a0s0e0d,o3n3t3h3aenfdin5d0in0g0sopgf. thetoxicitytest,themaximumdoseplatedinthemutagenicitytest was5000 igprplate.
Mutagenicity Test
inTa"Tbhleerse2s6uasnodf2t7h.eTmhuetasgeednaitciatwyetreetgeanreeprraetseednitneEdixpneTraibmleensts6tBhIraoungdh2B25.aTnhdesduomsmealreivzelesd
~
testedwere 100, 333, toxwiascobisertveyd.
1000,
3333and 5000
gperplate.
Neither
precinopriapptreacitabele
oftIhentEexstpeerrsitmraeinntsBin1e(iItnhietritahleTpersets)e,nncoepoorsatbisveenmcuetoafgeSn9icarcteisvpatoinosne.swereobservedwithany
obseIrnveEdxwiptehrainmyoeBfn2tthe(eIsntdeerpsetnrdaeinntsRienpeeitahterAtshseapyr)e,snenocepoorsiatbisveenmcuetaogfe$n9iacctrievsaptoionns.es were
CONCLUSION BacAtelriaclriRteevriearfsoerMuatvaatliiodnsTteusdtywwietrheamneItnadsepdeensdcernitbeRdepienatthAespsraoytioncdoilc.aTtheetrheasu,lutnsdoefrtthhee a`bcosnednitcieoonsfoAfrtioscsltuodyr,-Fi-r2na5td3l3ui9vcedrie$d9d.notcause apositiveresponseineither the presenceor
--_ Bs ioReliu anceOEave-
Company Sanitized. Does notcontainTSCA Cal
- Dc. Reverse Maton TestwithanIndependentRepeatAssay
DuPont-10819
REFERENCES
AmesM,utBagNe,nsJ.wMitchCatnhenaSnaldmEo.nelYlaam/aMsaamkmial(i19a7n5)MMiectrhoosdosmefoMruDteatgeecntiicnigtyCaTrecsitn,ogMeuntsatainodn
Research, 31:347-364.
Green, MHL.and W.J. Muriel (1976) Mutagentesting usingtrp+ reversion in Escherichia
coli, MutationResearch 38:3-32.
IntoemfatPiohnalaCornfmeraencceefoounrHtaHruimmocnainasaUtlsioesn.(GIuCiHd)aonfcTeeochnnicSapleRciefqiuciArsepmeencttssfoorfRReeggiustlraattoiroyn `GenoTesttsfoorxPhiarmcaceiutitcalys. S2Adocumentrecomfomraedopntiodnatestdep
4oftheICHprocesson July 191, 995.FederalRegister 61:18198-182A0pr2i,l 24, 1996. InternationalConferenceonHarmonisation (ICH)ofTechnical RequirementsforRegistration
`ToefstPihnagromafcPehuatrimcaaclesuftoircaHlusm. aSn2BUdsoe.cuGmeneonttorxiceityc: oA mStmanfdeoarrndaddBoaptettieordny aftorsGet no4tooexfitcp ihtey
ICHprocessonJuly 16, 1997.FederalRegister62:16026-16030, November21, 1997. Maron, DM.and B.N.Ames (1983)RevisedMethodsfortheSalmonella MutagenicityTest,
MutationResearch, 113:173-215.
OECD Guideline 471 (Genetic Toxicology: Bacterial Reverse Mutation Test), Ninth
Addendumto the OECDGuidelinesforthe
`Paris,February1998.
Testing ofChemicals,
published
by
OECD,
Vogel, HJ. and DM. Bonner (1956) Acetylomithinaseof E. coli: Partial Purification and `Some Properties, J.Biol. Chem., 218:97-106.
--_ sBuesylo.eSU
1s
------------------ Company-- Snes-- e0D0o2ss-- NoOttCONE-- IcripNTSCA-- CEI --
A cc. ReverseMutation
Test with an Independent Repeat Assay
PBraecltiemriinaalryMTuotraiticointyTTeesstt
DuPont-10819
Table 1
TESoxsetpaeyrSiutmbeesnmttbaneNcroe.
1a: |
S 525339 --
VDSeaahtieicnlPelates Dw1m0aee0rum 2002
TCEeolsnatcueinsntugrbAaslttiiagonnqceuor:Rev0We0irtihant5s9 |Aeptaisvkagvrioosnnd 00RW0eivtehrosuetnte39 Acstaicvhaats0ioou0nnd000
-pgpeVernpicliaete 1 perplate La3 wn 1 perplate LaT wn
3356
i11 ii:
3I1o iI1
51&30
ibbsod ih3
ibbds iii
--_
33Sio3o3o
b2id iI3
ii i
3I i
FacskoSez00oo0und Lawn Goa ds i ai
aa ai
NiTee<slBtlxootrnrmeaIlnnsctley2r-fSree1rdiiugnchgeedl:DyreSrceoidApubicateeadn;tte;;d-SlI-iomo-sdicenrataretesedfloyhryirnepydrupecrceeidcpiiptiattaete
--_----------
sBtiuodRyelNiaonce
1
CompanySanitzss. Doss no corgi 150A Co
I estwilh a I=ndepenridce)nRtevepressetMAutsaction
Dupont10819
RI ostra RY
bie2
J ----
Str udy e umber.: SEE
[Teoery TR LaJ s
mTenteSumbeeta"nce RevWeiitthimt5y9 RevTiviarimon, mWaios5398AheArniotairmitwioon,
ugperplate perplate Lawn
perplate
Lawn
r T eRRray tsL Sooetaiamhetlst y Srodse ceeds Ie tntfeodnaroeattetbydyzaeduporndedcloeteate--
BsiuooReyliN:E o EN
CompanySantee. pos conanTes co
~
SP: ccc) Reverse Mutation
"Test withan IndependentRepeatAssay
DuPont-10819
.
P[retinAimary-- Port-- e Test
bie 3
nese substance 10; ncasass
frSSPitsoutidieyyetMusshberre. H|e |E 1am 2S 0m
TPaibaiteiinsg tiger | | Foo
CToormtcecmorbaettiaonnca Revietrhi8e poBvievcairpiioonwd NWeivseshseasnts5 ae"rmievsairrieoenmd
Venice tore riste. ie pecplste 3Jwa.... 7 gerpise... 3lod...
ofsr
iu3 :ii
5s: i3:
f38oo
M8H 3ii
iiB iii
-
8ifo00
iii ii3
i8 iii
pioTr ohccmaly y SCeRighti2y
xeduced;
i SeModezataly
i sodiced
i
NieoomveoreInait1byceruriinnegd)beeSsaipbieoatne: sT-eoommsmcieerretdorbiyosprperceicpiiptiattaete
suyNo. EEN
Company Satz. Doonotcontain 50 Go
WcReve. rseMutation
Testwith an IndependentRepeatAssay
DuPont-10819
PBraecltienriinaalrymTuotxaiticointyTeTsetst
abe 4
mest. substance xa: 5-25339
StESrxtpuaodiryninNeunstberNo. I| S AmLasU a ES
&
DVeahtieclPelated |1 v1e8tuern 2002
Plating Aliquoe 100 5n
CTeosntcensturbasttiaonnce uo per plate'
|RevWeirtthan$t3s per plate
ActiBvaactkioornowd fan
RWeivtehrosuatnta$5 AoBraicvkagtroiuonnd per plate Lawn
Vebicle
To
T
10
T
b6t)1
13
1I
.5
1i
10F&o0
na
11
7
1
2a
11
.
1
io3r
a
iI
5s
iI
--_
S10033003003
8`
iI
s
i
:7
i:
:
1
SachgzTouenodrnLaalw;n 2Go-d1sightly iExtrenely reduced;
SreedAubcseedn;t;
dsGM-oOdbescruarteedlybyrepdruecceidpitate
Neston:Tntaxfering precipitate; IP-interfering precipitate
BioRelmee pp o
Study No.SY CompanySaniized. DoosnotconTtSaCAiCnB
Pe -- Test withanIndependentRepeatAssay gaia ein ner
S---- J Tyme StudyNumber EEE
DuPont-10819
Ta ER ayrm ,
bgperplate perplate lawn
oD
per plate
Se
Lam
eBE eeToneg tS tERt mdrr.y
-- Sa-- ye-->------ Company santos bos orcr Toon co
--_
-- } }
seikeeinn meron meee "Test with an Independent Repeat Assay
DuPont-10819
sain5
e 207% gatas T0y Brin1n0 01
Hone i
ERE
0 PS Clonacta0 inne eagaAtli0 ioqnuot0 TI:at2fe0. 00 wSLesee0 rieanta0 mnd0 Riagrlon0 d ew0 erwe0Lse0 ameTers0
w Sprroa8 d aw om E entE gelicstSEimEe e-or mmenbeyedepereciaptleente
E sudo ENN y
--_--
cComopasnyeSantas. oDooemsmoticconotmaimn5moAagotm
~
I osn os ...cRe. versei Mutac tion
esas
S IET a seriesJmonpi yon 3
Hsp | 20
D Torrnfa r vem r
senting tes Rgptimalgpgoe tg 0 sv TEEth Th2 elon asvayyssemnns saene
- -- EE
-_
eeEsseeeTseesoamse
~
"Test vith an Ini dependenetRevperatMAisssiyo: n
Dupont 10819
Bacterial witation Test
nabte s
Smseersatasunbsetance 18:-- n--as33s Liver Wicrososes | ona
DCeactre HiSaetnsneeddx"o || 531saux 2100002
VFaecnienrga iguor jp Joo
iCgonpcaerntprlaattieon vecle
Fuimebveer on
pReerveprltaanttes ise
Be1cCkogdraomd
seAvveerrtaagmec|s bSetveineitariden
sa amom 1i
0
00
aaowm am iia
1m u
FY
aa0 aamw eid1
1=
-
2000 sa Gaommadia
10
sam
e a omm m e aa
224 ,
sooo aaEO w ae i
=u
Rosiive Contron soooadiumoasimdeet1.0 4g peOTr place
TackgroTuenodrmLaalvsa
caovdSeiGigheely reeduced;
3 Jeboderatly
ons seduced
un
NioorBmxoinr_aInnacleyctsoedryicnegdspreSceiapbirteantt:es GTeroomimaetnerreidebryinpyrseceispiiptiavtaece
LS
--
sBioyRelNioa.nc( e I
2%
Company Saitzsd. Dossnot conanTs0A Co)
-
ove iasion
Test with an Independent rl Repeat Assay
DuPont-10819
SP rues
sTBtudrySvumrberm j|GEieEver 59 SSolien greiNaons3esxe
a ET Bn, Te wet ee----------
~
sag
SR ARO SY SeT E EA Ra IEo Im,iAnRhg SO
ermine -- Suro
A.
ETeEstEwith an InsdeopecndieRntevReerpseeat AssayMustion
DuPont 10819
Bacterial Mutation Test
able 10
ZsSeesrataiansubnst"ance 1a: -- -u-2s333 CsalolscSeieded10 |+ 2.2 X 10
LVievnesreloMicrosones Plating Mique
|}vaNevneer | 100m
Date' tiated | 3 aul 2002
Conpceerntprlaattieon lPulmabteer Rpeevrerptlaanttes BacCkoptrao.wmd HAevveerratyeent|s Sbetvaimeatriden
Vehicle 6a021 nnoo od1
1
5
100
0aa nnnooa1i
12
1
a
aaa Fnn oRo a
1
2
1000 oa0r ni 2 ooia
12
1
am
aa0 L 0 woi a:
prs
.
s000 oo0r 2F2 r 3a
n
:
Positive Control soaordium azidew as'1.0 a ug peRYr plate
Sackorouma Lawn Codea HA
a2
NeIinsotteoiorrnmeanTle;vley2r0fsSeeirdgiuhnctgeldypreSrceoidhpubicsteeadn;tte::dsoI-lohobdeseicrnuatrteeerdlfyebryriespdrupecrceeidcpiiptiattaete
P BioRele iancs e------------------p----
StuyNo. (EE
~~
I ccevr Motaiion
Test with an Independent Repeat Assay
DuPont-10819
Bacterial Watation Test
rabie 11
SSTototur.deytsNubustdaneces1:a:so (5.M2S53N3S5 SNMNNEE CoSlptesrisseersttedNo |: 2312 x 10
FVCiaiavnteiirangMiAclrioqsuomoers
}|toRaesiver | Tob
59
Date Piaed | 3 3u1 2002
bCooncpeenrtrapltatieo'n uFmlbateer peRervertplaantet.s secshagreowd neAwvveerrsaisasr|s Ssteamniaoaarrdd
Venice 3oor $nn0o0ni1
u
:
200 G5oan nnoai01
u
2
53
G0 5PPR 01
51
2
:
--
2000 oaar Fn0I oia
10
o
5333
aGa n0 wo1 ai
n
2
so00
aaa 0 nuo1 ai
0
:
Positive Control 2oo-raminoanchraSs iceenei1.0 yg per plate
w SackoroLuenodrmLaawln CaoedSehoichtly reduo ced;3 idw olederately redw scead a `
Nieeemheornr-TennteeyxfsaerdiuncgesproSceinpbisteantte:: STrpoebIemcceercefderbiyonprpeeceibpiiptiaotaete
Pr-- as--------------------
SBitouRdeylNioan.c
A C sciE Revs sain
Test with an Independent Repeat Assay
DuPont-10819
rot smn 10 msraioniemnmem or StudyNusber :QESSSNSNNMMNGNe ~~ ScoerimeNnot : BL
Ce= Sotmeemoernontse sTa-- tte43e4Serer erE Ho erereN Sa kgper plate Number perplate Code RevertantsDeviation
T TEREnT5 TSSEimyetyB ereseentt sb Severpsceyera ennee tw Sirs a Seedy peeiriee ESE ra soit.QE
Cone So re ogc co
-
I ci Revere Mion
:
Test with an Independent Repeat Assay
DuPont-10819
[----
rabte 13
TSroeesta.tsaubmsbtearnce Ta:G| --5._E25--33N9 ER Eiver Microsones | nat 2ves 9
ECSaptseeciPSaoevenetddsv"o :| 3E21zax 1o0r0a
VFiennceirneg atiauor | e10s050
Cosnpceerntrpaltatieon Vemicie
NTuimabteer pReevrerptleanttes
01
BecCkogtreowd mveveerrsesoests ostamndearrd
oa
ii o a
s
:
100
a0a2
PPsooadd
`
:
ss
oa&
PFEE i
.
:
--
1000
G=on
[o A da
.
3
sas
@aa
i HE aa
,
:
so00 aaa woC sd a1
,
s
Positive Control 2&-5aninsanthrs aseceensd1.o0y ug pe plate
-- SacharToouhnoarmTaalw; CSo ClSelaightlyoredsu5 ced:1 3 Seoderately
ow2 reduced
a2
NCooeihroenn-TtattyeteadruicnegdsProScoinpbieveentte:r STeooebteocteorretdobriynneomtepaititmattieate
--BioRrelian--cs=--eeeereserereemememmeremeemeseeeee.
studyNo.(
a1
~
est with anc Indee pendenRteRveepresaetMAustsataiyon
DuPont. 10819
Bacterial Mutation Test
Table 14
TSteusdtysNuubmsbtearnce Strain
1d::QH-E2S53S3S9ENSNNBY S22 wn
CoElxserSienedeeNndot :: 3B.L7 x 10
VLeihviecrleMicrosomes : Naotneer
Date Plated : 2 ul 2002
Cugoncpeerntrpalattieon PWluamtbeer pRaerveprltaatntes BacCkosdreouwd RAevveerrtaegnet|s SDetvainadtairodn
Veicle aor 1100 11
0
un i
n
2
100
0002
11s pri
11i
1
1
EE
0a
n1u 31
03
un 1
1
2
~
1000
0a12
1un 11
0
ui
1
1
3333
0oz
11 11
03
1I
1
2
5000
00
11 11
I is i
15
1
Positive Control me0t1hyl met1ha0n3esulfonat1e 1000 ug per plate
-- o Background Lawn Code003 %a 1 s ii s ss y5
NPIdsn=lNEioxortnmra-elIn;netle2yr-frseelrdiuigcnhegtdl;pyreSrceeidApubicsteeadnt;te;;36mI-MPooebdisencrtuaertreefdleybryirnegpdrupecrceeidcpiiptiattaete
`BioReliSe-- ancs
~
c Dleessttwwiitthhaanns IInnddeeppeennddeennvttRReepepeeaattAAsssssiaoyn
ay
Dupontiogty.
Secterted Hermione
DuPont-10819
mite 15
Test substance 1a: nasa
Souay wber : GUNNS
StEtaorvaeisnMicrosomes |I: atmWP2 vuevezaAe 3
er BxperiYon:en5t1
Cells Seeded : 3.7 X 10
CosFnipcnoetrnitnprglaatntireoinqueeuFmiab|teeYrop pReervepriaatnets secsiagreomd pvaeerresiens,, sreaesntdearrd
Venicie 23ore #woo 01
1s :
100
2a
& io
u:
I
aGo Hw oo a01
s'
1000 wa
~
z02
#14
i1
ws
:
su
3"a #w FA 0a1
"3
s000 aa u 50a1
Rositive
Control.
2a s-pamnitnnoomaE nnethheieanecene
1100
yg a
per
plate
SacrarCToeuoRnEodrrmeTw ameliywtyCaeoSeo eehv bimsehettlsyz m Sreodupcbedm;eF 3oeNpoiedrearaetleyly rTs eedduuccseeetd rate we
NEemmnon iotecstating prectbiete: Tocmmteeaerer Ae
GeFioRr etnce eeee.
sue. GEE
~ WE -- rere mmm an Independent Repeat Assay
ee DuPont-10819
RR Be ody Nude: :SSN FE hr. EporizNoe:n2t2 rTre I ten vem ae em
~~
02
19
1
Poison ten testes 3.0 ng sine E E STREELcnnm esenyt,E ceetnaosu ees
-- a EE ----
~
Teestswtitih anIdnedpeecnpcdceennRtRdReeeeppveneeaatttMAAusssssasaiyyon
DDuuPPoonntc.1100881199.
Bacterial mutation Test
Tate 17
Sy Sohreaty hp ier | S--- S
CDooilsssiSeeasdevdo | 33.4 x 10
VLPelinavitecirtnegMimcrioqsuoomres
| |
R1w0aa0tverliver
$5
Date Placed | 1 ut 2002
C1o9npcoenrtprlaattieo"n Vehicle
NPulmabteer pReerverpltaatnet's BacCkogtraowd naevveerratgeems bSeeteonraircde 01 wo 1
aoz
w FO a
1
1
100
ao0 nnn oo oai
1s
a
3
a0 a
2FnA oia
1
2
--_
1000
oaa FaL O oaa
1
2
am
aa02 wnnooa:i
pr
:
s000 oozr o
w FBr 0a1
n
2
Positive Control 20a-aminoanthrFm aEcensa1.0 yg per plate
BackorIoeutnldorLnaalm;
Codea 201ighely
Fer reduced;
Ietodorately
19 reduced
NiPeeBNxotnreInnatleyrfreerdiucnegd:preSc-ibpisteantte; stePomienctuersefderbiyogprpeeceispiiptiaitaete
_ BioRelance --_--) sayNo. AY 35
~
estwithanInde-pe.nd.enRteRveepresaetMAutsastaiyon
DuPont-10819
Bacterial Mutation Test
Table 18
STotsutdysNuubwsbtearnce strain
Id:: QEE-E35S33S3SNNNMMN + Too
~~
CEexlplesriSneeesdtedNo
::
152.4
x
100
Liver Microsomes : None
VPelhaitcilneg Aliquot : w10a0terut,
Date Plated : 1i dul 2002
uCgoncepenrtrapltaitoen PWluamtbeor Repovrertplaanttes BacCkogdraound ReAvveerrtaegnet|s DSetvainadtairodn
Vehicle 0o2r 221051 11
03
167 1
19
25
100
o0r2 115788 11
03
185 1
107
10
.
333
0012 a2637 11
03
201 1
210
2
--
1000
o012 1c92 1
03 152 1
152
o
3333
o0r2 220108 11
03
20 1
206
s
5000
oaz 221050 11
03
27 1
am
5
Positive Control s0o1dium azide5741.0 ug pe1r plate
-- 8 Background Lava Code00 5o6n3 3 s 11 e 603 msss
NdLp=e=BNNxootmnra-elIn;netle2yr-fSreleridiugcnhegtdl;pyrerSceeiAdpbuiscteeandtt;;e;3seIMGpob-daIecnrutareteredfleybryirnepgdrupecrceeidcpiiptiattaete
Cecontaninated
:
sBiwoReeliancQe E
eee ORIGTSCOCR
~
A sc. ReverseMutaion
TestwithanIndependent Repeat Assay
Bacterial Hutation Test
DuPont-10819
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ProtocslSPGTSO001 06Jm2002 Pa7gof1e1
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Protocol SPGTS0001 06Jun2002 Pa8gofe11
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120 REGULATORYREQUREMELNABTORASTO/RYGPORACOTIDCE
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Rveigtihstehuionn omfP inConu tefso00enHm aumUossei(s1i9o%a n nodf1T9e57c)hie Regainsor
iPTriipsrcosceteusdRyepwhaiallslieb,oo nphesecrfvomredNdco,nlncdoirmeppLrriev)wriwsitlhnl ybhoSe pturooeviicsdi(opnGesLtfPhh.SeTaGhnoeodpdsrLipobetrrsoiro,yr
Pcirnoopmecpcetldiiuoncncsw(wiSilOtlFhGb)oLoPpfse,BohaerRSesOldPatsnooc.fbBydtiithelQheuicltiayaAlscsheuryaccysFopUenricothooafut.BsTisoooRidcnipcrioecefcoss
sspSspoyccpiin twcihlplahsLasssye.t,hAethsedidgenoendsQpofAsesscthunppetccvteiidl d,booiafngottihhee.dpiaressviotohufesroqeupasecof.5etTxhhihs
iandsmheeiccoainnotnwwiseilglrnseetdpGoLhrPeceGoLomtPphleuikSedtsisynDteie)emtwislhsiwdlhitbcsiStnehncdsheDydiins5thceorfmopliangretpeaoretd.nsThIyins
, eoxrcepbtilonylolthyisecoomfpolitoae,ofcsopeldieti,nxesaonbie rtiron.of oacersion
ProtocolSPGTS02001 06Jun2002 Paget 11
@ BioReuANCE
t BioReliance eta bomens eesti
sudyNo. SEN
57
----
.
--
I sci Reverse Mutaiion
Test with an Independent Repeat Assay
DuPont-10819
`SponsorProject Numbec: _Dabont-10819
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10 REFERENCES
Re2Amssc,hBgNeS,nsrMwail6mh8,ho .SoandomYaama,TEa.l(197)r. oMtmheodMs iAGdeScitocicHionwogseoens
EOcrheri,cMiIsLcLoi,laMndMiarksl,eWsiaa.o3(319.7n63,.Musgen testingusingtp"rovesicn i
RReeIggtiletrtaaotiriooynnGsoesfCeoPnrhfiarcemtnaycceeTseosnisdHsafromProhsaiHmesanicoensUi(scCenH.G)uSlo2fd4aTdeoseccu0mh0ennStprRicetogemiiAmneepnietesdsfoorf
adGpUioTnSa938ph1o4r83o24f,01h9e26,KHpson ly 19, 195, Foden Regier
~
IGeReergotiasirooaiinocnayoCTfoPensafiragesosfceePuorinaoaHscamoearssiicsns.inSo2nU8(sd.CoGHu)amoetftoTesccoh:mimceAanSltieRdedgbosoieBmoueapctisy boo c
ie n CHe procesonJl16,197. Fede Regt6160361680, Newent
SeMaclCrnaonnnel,1l.issnnidcArvesso,emssBtXs.s(1o97f),0D0ectshcainoenlo:fcGesusaionPnrs oNatgeeBsenidns . Shee
SMaetCUaaStnA, 12,5Ch1i3so5S,51Eo3,Y5n,easlsi,B.sndAekm.s,0BN.o(f1290709c,aDmiesoofcet.ioNgeoteaesnn3t
MaTreon,tDaMi.annRdeAmaers,BN,11381933)3,5Revs Methods otoSonesMey
PAOudBesCnFdGamriy0dh1le9O84nE.7C1(DGGeuindtlsiTnoexsicoorlo5gyT:BoahtoecoflCRheveemriseeM,optiiloenTde)5,ONRiCs,h
SSWialuens,aiP.pNnhaiiomdonoA,lTAWlGeEb,iDJ.c sdGh atoe s,WPDGr. e(1s9si 90)t,iC.os mWpsarsipsh noncsosf
Protocol SPGTS02001 0632002 Page 00f11
@ BIORELIANCE
-- BioReliance
Stuay NoSY 8
~~
ci vseMain
lest with anIndependentRepeatAssay
DuPont-10819
140. AprROVAL
B Spoo sarst tjyc Nus mber:_J DuPen 0819
USa pooe r ReprDaionenpf
10 dbume 2002
TTMoereian Donner
-
NolBieRcltanicesS.y TC
retK
Res SoyMag
13 T=an 200m.
13 yun 2002
ew
Probocol SPOTSO2001 06402002 Page of11
BioReliance
Study No.SE
2
EE
--------------------------------
@ BioReuANCE
Se,
A Testwith an IncdeipencdenRteRveepresaetMAustsaatiyon
DuPont-10819
APPENDIX C Information for Japanese Regulatory Agencies
BoioiReleiancE e E
ee RSAZ e DlCOUIBIOTSCACE
~
est with an ccIcndependRenet RveepresaetMAostsaeion
DuPont-10819
RepofoRersultts of Reverse-MTeasttianBtaciteornia
1.TesterStrains
1) Procurement
Obisined from |Dateobained Dtahteesitnmspelctoeitdn
storage
10November 199
:
TALS UDnri.vBerruscietAy mofes | 1 August=1998
[rm] oi Ja TALS
ele
-- 14November 1990| con"figrmeedcoen the
~
ationOTon
(WkeM2IuOv1r)A oMafrIinnduesBtraicatlearnida Huyiss?
[sempLarge qr uently e]
Coommppoossiittion
DMSO
0.09mL
BioReliance
study No.
C
--_-- iSecd-- ceeTaca
~
E "Test wE ith an oIndcepecndenRteRveepresaetMAsasiayn
DuPont 10819
2.59 Mix 1) Source, StorageTemperature, etc.ofS9
re[one Purchased fromMolto | Prepared
Preparationof $9
ty 18 April 2002 (Lot 1384)
J | 14May2002 (Lot 1390)
SO r ES n Speciiee, s, StraiStrainn| Rattus norveggiiceuss, Sprague.
Administration
5 (Batch R6G3)
unknown (Lot 1384)
Aee@wesks)| ynjnown (Lot 1390) | Administration
21110251 g (Balch R663) | p(egrkigo-dweaingdhatm)ount
Weight
uunnkknnoowwnn ((LLoott 11339804))
Aroclor. 1254 noah
single dose at 05 A weight, sacrificed after
5 days
S BioReliar ncs e--
eee CompA any SSaIriRS D020CRRATSCA
~ Ac ReveMision lest withan Independent Repeat Assay
DuPont-10819
[Tn [E--E] 3. Preparationof Test Substance Solution
`Water (CAS No. 7732-18-5)
Corporation
1129947
Distilled
or`Stability of test substance in
`Methodofsuspension when test substance is difficult to
| Not applicable
Hvri
suo EE.
-- -- EE ------------------A -- a-- R e-- lGota-- nTSCA-- CEL --
~
a Bacterial Reverse Mutation
`est withanIndependent Repeat Assay
DuPont-10819
Te |oe[seer] 4. Conditions ofPre-culture ue| os oietooc ntgiamr irgot [|f21i05onsmstmin empemrs | Suman
eSntdoorafgce tuimelatnotdustueefmpor.rfteresotm |<8 hoawt2s8ooC
Model and manufacturer of New Brunswick Scientific, model G-24
Me(thsodhotfyasphea,kksipniege.dn,etgc)_|R`RotaryO (1(12235 eviremv/imisn.)
~
JCuoltourse vessel (shape,
shape:. cylinder,220000mL
-- Suir e -- A ---------------- Competing ores canca ca
P sc cc io `estwithanIndependentRepeatAssay
DuPont-10819
5. Agar Plate Medium 1) Top aga
Agar [Loo |
2) Minimum Glucose Ag:
[Mme 1 Agr Volumeofagar plate medium
woouksq mhisae | 100013DKSQ
--
6. Test Re-sJudugemlenttoftshe results.
[men [Neve
Reasonfor judgement and referential matters:
ines of Arlo mdneod res 00 No positive response was observed with anyofthe tester strains in either the presence or
Referential matters
`The vehicleandpositivecontrolvaluesindicatethatalltester strainswerefunctioning
correctly and were capable of detecting a mutagen.
~ Ee dm NE
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