Document YD4RZ9kz0KoRvZdbZYzY2L10
AAG. 3230
412
ARRRG - 3230
DuPont-8868
TRADE SECRET
Study Title 'H-25171:BacterialReverseMutationTestwithan IndependentRepeatAssay
Authors
Valentine O. Wagner, III, M.S.
Michelle L. Klug, B.S.
Study Completion Date:
11 February 2002
Performing Laboratory
BioReliance
9630 Medical Center Drive
Rockville, MD 20850
for
E. L du Pont de Nemours and Company
--
`DuPont Haskell Laboratory
P.O. Box 50, 1090 Elkton Road
Newark, DE 19714-0050
PerformingLaboratoryStudy Number
'AAS2XL.5020B0T1L.
WorkRequestNumber
[my
{mm `Service Code
py
Page 1 of 62
HA25171 BacterialReverseMutation Test with
an
it
Assay
DuPont-8868
STATOE FCM OME PLIN ANT CE
Study No. AA52XL.5020B0T1L was conducted in compliance with the US. FDA GLP Regulations as publishedin 21CFR 58,theU.S.EPAGLPStandards40CFR160,and40 CFR
7P9ri2n,citphleesUofKGGooLdPLaCboomrpatloiraynPcreacPtricoegirnaamlmlem,attehreialJaasppaencetssewiGtLhtPhSetfaonldlaorwdi,ngaenxdcetphtieonOsE: CD
The identity, strength,purity and composition orother
characteristicstodefinethe test andcontrolarticleshavenotbeen: determinedbythetestingfacility.Thecontrol articleshavebeen
characterizedas pertheCertificatesofAnalysison file with the
The stability of the test and control articles has not been dete byrthemtesitinn gfaceilid ty.
Acnoanlceynstersattioonodfettheertmeisntaetndhceonutnrioflomrimxitturyes(waesraepnpolitcpaebrlfe)o,romerd
--
by thetesting facility.TheSponsorhasindicatedthatthey have
`not performedthese analyses onthe testsubstancemixtures.
"Thestability ofthetestandcontrolarticlesin thetestandcontrol `mixtures,respectively,hasnot been determinedbythetesting
facility.The Sponsorhasindicatedthattheyhavenotperformed
stabilityanalysison thetest substancemixtures.
Noted O-Wogan,TC SVatluednytDiinreecOt.oWragner, Ill, M.S.
eretX Y
BioRelianceSnady Management
IL Feb 2003
Date
11 Feb 2002
bw
BioReliance
--_
Study No. AAS2XL.502001.BTL.
2
Company Sanitized. Doss notcontain TSCA CBI
25171: Bacterial Reverse MutationTest with
anIndependentRepeat Assay
Quality Assurance Statement
DuPont-8868
StudyTitle: RE12P5E17A1T: ABSASCTAEYRIALREVERSEMUTATIONASSAYWITHANINDEPENDENT `StudyNumber:AAS2XL.502001 BTL StadyDirector:Valentine O. Wager,I, MS.
e"GATohiornsdstLeusidbyohmraeiosonbreyGkePnasridacicnviocieheddeRodenignunctloeastaPrisoaenrsssies(ss2o1ofCivFni-Rrp.r5ocb8see,sssOhpelhahyUsSeis.s.pEcUPsoiAnlnGgLodaPsrsea(ns4ds,0omcdCsoFaacmRurp7pl9eei2nnaswgniiaodplnp4r,h0oCeaSUcFuhRp,S1mQ6ueE0anlD)ti,Attyhe UnKdtGyLPsRoeugalhatsotnhse tohedyJ1aspacnoensdeuGcLISatcacnodradridn,gandtthheeproOtEocCoDlandPrinrcipeleslevanotfGStoaonddaLradbOorpsetorryaP.ractice "Thefollowingare the inspectiondates, phasesinspected,and report datesofQA inspections ofthissy.
PIhnsapseectOn lPahsapseectOn PIhnasspeectOn PhlaasspeectOn
P0r6o-tDoecco-l0R1ev-i06e-wDec-01 To StudyDir06-Dec-01 ToMgmt06-Dec-01 2S4c-Doet0rh1ei-p2ln4at-geDsec-01 To SedyDis24-Dec-01 ToMgmt 26-Des-01 D0r3-aFebR-e0p2o.r-t03-Fe-02 To SyDir04-Feb-02 ToMgmt07-Feb-02 1D-Fertbo0Fai2n-alf1R1-eFpteobr-t02ToStadyDir11-Feb-02To Mgmt 11-Feb-02
Thisreportdescribesthemethodsandproceduresusedinth studyandthereportedresultsaccurately reflect
therawdata ofthestudy.
iy
Leck D. Schweckeugat
BeQcUkALyID.T/YScAhSreScUkeRnAgoNstC,EB.S.
II_Feb 2002
DATE
BioRelance
--_
Study No. AA52XL.502001.BTL
3
`Company Saniized. DoesnotconTtSaCAiCBnI
Ha.n2I5n1d7e1p:eBnadcetnetriRaelpReeavteArssseaMy utaTtesitwoitnh
DuPont-8868
CERTIFICATION `We,theundersigned,declarethatthisreportprovidesanaccurateevaluationof dataobtained. from this study.
IssuedbyStudyDirector: NateVialteintninee OO..WWaaggInILeMeSrn., I 11 FeDbate2002
Approved
a--
ee`Maria Donner, Phs.D.e. Senior Research Scientist
08-Geh ~2002
Dats
BioReliance
-
Study No. AAS2XL.502001 BTL
4
Company Sanitized. Does notcontain TSCA CBI
H-25171: Bacterial ReverseMutationTest with an Independent Repeat Assay
`TABLE OF CONTENTS
'DuPont-8868 Page.
Characterizationof Test and CODLIO! SUDSIANCES........cwwusmmusssssssmssssssssssssssssssssssnd.
FO
----------------
Table 1: Preliminary Toxicity Testin Salmonella typhimurium TAB........c.owwvvn:18
-_
`Table 2: Preliminary Toxicity Test in Salmonella typhimurium TAL53S ........oovwwnr19
Table
Table
3:
4:
Preliminary
Preliminary
Toxicity
Toxicity
Test
Test
in
in
ESsaclhmeornieclhliaatcyoplhiiWmPur2iuVm TTAA1S.37......covvvvreri220
Table 5: Mutagenicity Test in Salmonella typhimurium TA98 WHhOUL Serr 22
Table 6: Mutagenicity Testin Salmonella typhimurium TA98 With S9 ...o...cvvivin23
Table
Table
7:
8:
Mutagenicity
Mutagenicity
Test
Test
in
in
Salmonella
Salmonella
typhimurium
typhimurium
TA100
TAL00
WWiitthhO8U9t
$.9......c.....e..r..2n254
Table 9: Mutagenicity Test in Salmonella typhimurium TA1535 without S9 ..............26
`Table 10: Mutagenicity Test in Salmonella typhimurium TA1535 With $9 .....occvcvve2en
`Table 1: Mutagenicity Test in Salmonella typhimurium TA1537 Without $9 ...............28
`Table 12: Mutagenicity Test in Salmonella typhimurium TA1537 With $9 ......c..covvern29
`Table
Table
13:
14:
Mutagenicity
Mutagenicity
Test
Test
in
in
Escherichia
Escherichia
coli
coli
WP2
WP2
uvrA
uvrA
Without $9.............v.r..30
With S9...........vww:3L
Table 15: Mutagenicity Test in Salmonella typhimurium TA98 without $9 ..................32
Table
Table
16:
17:
Mutagenicity
Mutagenicity
Testin
Test in
Salmonella
Salmonella
typhimurium
typhimurium
TA98 With $9 ........ovvoven33
TA100 WithOUt S9 .........34
Table 18: Mutagenicity Test in Salmonella typhimurium TA100 With $9 ................35
"Table 19: Mutagenicity Test in Salmonella typhimurium TA1535 without $9 ................36
RT"TTaaabbw bllleee 222012::: MMMuuutttaaagggeeennniiiccciiitttyyy TTTeeesssttt iiinnn SSSaaalllmmmooonnneeellllllaaa tttyyyppphhhiiimmmuuurrriiiuuummm TTTAAALL1SS53337S7 WwWiiittthhho$u9t $99.......3....]3.398 `Table 23: Mutagenicity TestinEscherichiacoli WP2 uvrA Without 89..............ccvcu. 40
--_
Study No. AAS2XL.50200BT1L.
s
Company Sanitzed. Does not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with an Independent Repeat Assay
DuPont-8868
`Table 24: Mutagenicity Test in Escherichia coli WP2 uvrA With $9...
41
`Table 25: SalmonellaE. coli Mutagenicity Test ~ SUmmary of Results BI ...............42
`Table 26: Salmonella. coli Mutagenicity Test ~ Summary of Results B2................43
ADPEDMiBx:SOUAY PROOCOL.....rremnrssssnsnses 46 Appendix C: Information for Japanese REUIBIOTY AGENCIES... SB
BioReliance
-
Study No. AAS2XL.5020B0T1L
6
CompanySanitized.DoesnotcontTSaCAiCnBI
-
H-25171: Bacterial Reverse Mutation Test with
an Independent Repeat Assay
DuPont-8868
STUDY INFORMATION
`[ SubstancI eTesi:NDY3
`Synonyms/Codes: | I
H-25171
)
HaskellNumber 25171
Co [EE mo
PhysicaClharaisisics [NSN]
Stability: Tcchohnesdetiretvsietodns.subosfttahnecesatpype:arweodetvoibdeencsetoabfleausnadebritlhye was Sponsor: EWU.LiSlAmdi.ungPtoonnt,dDeeNleawmaoruers1a9n89d8Company
`IStnuidtyated/Completed: Decem05b,2e00r1/ (see reportcoverpage)
In-LifeInitiated/Completed: December20, 2001 / January 22, 2002
_
BSitouRdeylNioa.nceAAS2XL.502001.BTL
7
Company Saized. Does not contain TSOA GBI
H-25171:Bacterial ReverseMutationTestwith anIndependent RepeatAssay
DuPout-8868
SUMMARY
Thetestsubstance, H-25171,wastestedintheBacterial Reverse MutationTestwithan IndependentRepeatAssayusingSalmonellatyphimuriumtester tcains TASS, TA100, TALS3S and TAIS37andEscherichiacolitester strainWP2wwrAinthepresence andabsenceof Airnoccorlpoorriantdiuocnmeedtrhaotdl.iTvheerfSi9rs.tpThhaeste,etshtepwrealsimpienrafroyrtmoexdiciintyttewsto,pwhaasseuss,edutsoinesgttahbleisphltahtee idonsdee-preanndgeentfroerpteahtetmeusttsa)g,ewnaicsiutsyedtteosetv.aTlhueatseetchoenmduptahgaesnei,ctphoteemnutitaalgoefnitchietteysttessutb(sitnainticael.and
`Tetrahydrofuran(THF) wasselectedasthesolventof choicebasedoncompatibilitywiththe tteatrrgaehtcyedlrlosfuarnadns(oTlHuFbi)laittyaomfatxhietmeusmtcsounbcsetaatrnactei.oTnheotfaepsptrsouxbismtaatneclewy2a0s0smoglu/bmlLeb.utcloudyin
Inthepreliminarytoxicitytest,themaximumdosetested was 5000 ugperplate;thisdose.
wasachievedusing aconcentrationof 200mg/mLand a25 uL.platingaliquot.Thetest
substancewas aworkablesolutionfrom40to200mg/mLand asolubleandclearsolutionfrom
0.27 10 27 mg/mL. Dose levels tested were 6.7, 10, 33, 67, 100, 333, 667, 1000, 3333 and
-
5000 ugperplate.Precipitate wasobservedbeginningat 100 ugperplate. Noappreciable
toxicity wasobserved.Basedonthe findingsofthetoxicitytest,themaximumdoseplatedin
themutagenictietsyt wasS000ugperplate.
Inthemutagenicity test,nopositivemutagenicresponse wasobserved.Thedoselevels testedwere15,50, 150,500,1500and5000pgperplat.Precipitaewasobservedbeginningat 500 ugperplate. Noapprteoxiccitiywaasobbselrveed.
`TheresultsoftheBacterial ReverseMutation Test withanIndependeatRepeat Assay indicatethat,undertheconditionsofthisstudy,H-25171didnotcause apositivemutagenic responsein therthepresenceorabsenceofAroclor-inducedratliver $9.
BioReliance
a
Study No. AAS2XL.5020B0T1L
8
Company Sanitized. Doesnotcontain TSCA CBI
-
aHo-25171: Bactetrial ReAvserssaeyMutation Testwith
DuPont8868
PURPOSE
`Thepurposeofthisstudywastoevaluatethemutagenicpotentialofthe testsubstanceby measuringisability toinducereversemutationsa selectedlociofseveralstrainsofSalmonella typhimuriamandatthe tryptophanlocusofEscherichiacol stainWP2uvrAinthepresence. `andabsenceofAroclor-inducedratliver $9. AcopyoftheprotocolisincludedinAppendixB.
Thisstudywasconductedincompliancewiththetestingguidelinesofthe ICH (1996and 1997)andOECD (1998).
CHARACOTF TEESTR ANI DCOZNTA ROLTSUBISTO ANCNES
"i Thetestsubsta`nncuem,bHe-r2A5A17S1Z,XwLa.sTrheecetievsetdsbubysBtiaonRceelwiaasnccheaornac0t3erDiezecdebmybtehre2S0p0o1nasnodrwaas sa
iN
wsahsouplrdovibdeed.stUopreodn raetceiapmtb,itehnettetsetmspuebrsattuarnec.ewAansdeesxcpriirbateidoans of
`ity,wascsotmoproesdiatitornooormotthemeprecrhaartaucrtee,ripsrtoitcsetctoeddeffrionemtlhieghtetsansudbmsotiasntcuerae.ndTthheestabilitydBACIy,
--
ofthetestsubstancehavebeendeterminedbytheSponsor. TheSponsorindicated.that
`BioReliancewouldnotreceivecopiesofthesereports.
`Thevehicleusedtodeliver H-25171tothetestsystem wastetrahydrofuran(THF,CASH 109-99.9), GoldLabel,99.9%,purchasedfromAldrichChemical Company. Test substance dilutionswereprepared immediatelybefore use anddelivered tothetestsystem underyellow light.Toenhancetheworkabilityofthetestsubstance,thetopconcentrationwassonicatedin a s5c0reCwi-cnasp tcuberforetaupbpwersodx-uirmiacntgedlaoysi1p5nmgi.nutesatS0C. Subsequentdilutionsweremaintainedat
Positivecontrolsplated concurrentlywiththemutagenicity testsarclistedbelow.All positive controls were diluted with dimethyl sulfoxide (DMSO) except sodium azide, which was diluted with water. All subivided solutionsofpositivecontrolwerestoredat -5to -30C.
59
Concentration
Coe lt]eprere ---- T ]R (AldLriochtChNeom.i0c9a1l06CPoS,Inc.)
WP2 uA
ExpC.ADSatNeo.1641-3S-e1p3--28005
Purity >95%
BioReliance
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Study No. AAS2XL.502001.BTL
9
Company Sanitized. Does not contain TSCA Cal
aHn25I1nd7e1p:enBdaecntteiRaelpeRaetveArssseayMutation Test with
DuPont8868
|oe JB] meee] s9
"
Tash TAL00, TALS3S
(Aich2iCthoefmliucoarleCnoe. Inc) ExLp.oDtaNtoe.0088-7M0ar7-H2S006
CAPSuriNtyo. 650978-%57-8 (SigSmoadCihuemmiacziadleCo) lLoot N.o. 0o98oHOlI65? `CASPuNioty. 26569298.%22:8
Sop Concentration
(SLiogtmNao.Ch1e0mi6c0a6l68C1o)
_
TASS
ExpC.ADSatNeo1.89-N0o4v5-29004
Purity 508%
(AMledtrhicyhlCmheetmhiacncasluClfoo.naItncc)
Lot No. 15526A0
WE ur
ExpC.ADSatNeo2.66-6No2v7--23003
Putity 599%
muta"gTeonciocniftiyrtmestthse wstaesriplliatytoefdtohnesteelsetcstuibvsetaagnacrew,tihtehhaingahlesitqueosttsvuoblsutmaenecqeudaolstoltehvealuusseeddinn tthhee tes.Theseplateswereincubatedunderthesameconditionsssthe test.
=
BSitouRdeylNioa.nceAAS2XL.502001BTL
10
Company Saitized. Doss not contain TSCA CBI
H-25171: BacterialReverseMutationTestwith anIndependent Repeat Assay
DuPont-8868
MATEARNDIMAETLHOSDS Forsubmissionto Japaneseregulatory agencies,additionalinformationisincluded in AppendiCx.
Test System
TATI0h0e,teTsAtIeSrs3tSraainndsuTsAeIdSw3e7reasthdeesScarlimbeodneblylaAfmpehsimeufrialu.mh(i1s9t7i5d)inaenaduxEoscthreorpihcshiTaA9co8l,i WP2uvrA asdescribedbyGreenandMuriel (1976).Salmonellatesterstrainswerereceived directlyfromDr. BruceAmes,UniversityofCalifornia,BerkeleyandE.colitesterstrainswere: receivedfrom the NationalCollection ofIndustrialandMarineBacteria, Aberdeen, Scotland.
TesterstrainsTA98and TA1537arerevertedfromhistidinedependence (auxotrophy)to histidineindependence (prototrophy)byframeshift mutagens.Testerstrain TALS3S isreverted bymutagensthatcausebasepairsubstitutions. TesterstrainTAI00i revertedbymutageasthat mecacuhsaenibsomthinfraEm.ecsohliifits saenndsibtaisveeptaoirbasseub-sptaiitrustiuobnstimtuuttaitoinomnus.tatSipoecnisf,ircaittyherotfhathnefrraemveesrhsiifotn `mu(Gtreean atnd Miurioel,n197s6).
-
`Ovemightcultureswerepreparedbyinoculatingfromthe appropriatemasterplate or from
theappropriatefrozenpermanentstockinto avesselcontaining ~S0mLofculturemedium.To
assurethatcultureswereharvestedinlat logphase,thelengthofincubationwascontrolledand
`monitored.Followinginoculation,eachflask wasplacedin arestingshake/incubatorat room
temperature,Theshaker/incubatorwasprogrammedtobeginshakingat approximately 125rpm
at 3742Capproximately 12hoursbeforetheanticipatedtimeofharvest. Eachculturewas
`monitored spectrophotometrically for turbidity and was harvestedat a percent transmittance
yviiealbdliencgouantittteersotsfoanpnpurtorxiiemnattaeglayrp1l0atceesl,lasnpdetrhm.ildlaitliatiesro. Tnhfeilaecbtuutalntoittperrsesweenrteeddietnetrhmiisrneepdobrty.
R`TehveerssteudyMwuatastcioonnduTcetsetd)t,ocaodompptledy wJiutlhyOE19C9D7Gu(ipdueblliisnheed471Fe(bGreunaertyic T1o9x9i8c)olaongdy: wBiatchteritahle
Intemational Conference on Harmonisation of Technical Requirements of Registration of
PharmfaorHcumeanuUste(i199c6anad 1l997s).
Metabolic ActivationSystem
prep`aArreocdlforro1m2m54a-lineduScperdargautel-iDvaewrSle9ywraastsuisnedduacsetdhweimtehtabaosliincgalcetiinvtartaipoenrsiytsotneeaml.iTnhjeectSi9onwoafs `Aroclor 1254, 500mg/kg, ivedaysprior tosacrifice. The S9 wasbatchpreparedandstoredat "a7m0inCooanrtchorladceernuentialnudsed7.,E1a2-cdhibmuetlhkyplrbeepnaz(raa)tainotnhorafceSn9e.watsotesftoerdmsformiutstaabgielnitiyctotometSaablomloinzeel2l-a iyphimurium TA100.
-
BioReliance
-
Study No. AAS2XL.502001 BTL
1
Company Sanitized. Does not contain TSCA C81
H-25171: Bacterial Reverse Mutation Test with an Independent Repeat Assay
DuPont-8868
`gluTchosee-S6-9phmoispxhatwea,s4pmreMpaBred-imnmediiacteloy btefoiredniintuasculmesoetiiadneddphcoeosnpt-haaitanee,dd8m1e0M%nMSg9i,Cln5 maenMd 3c3onmtaMinKinCgl1i0n0maMp1h00osmpMhpahtoesbpufhfaetreabtufpfHer7.a4t,pwHas7p.r4e.pTahreediSmhmaemdSia9tmeilxybteufroere(iSshaumsmei.xT)o, `confirm th steiltyofthe S9andShammixes,a0.5mLaliquotofeachwasplatedon selective
agar. Solubility Test
dimeAtshoyllubsiullitfyoxtiedsetw(aDMscSoOn)d,ucettehdatnoolse(lEeOctHt)h,e vaecheitcolnee.,Ttheettraehsytdwroafsurcaonnd(uTcHtEe)d,usaicenlgoniwtartielre,,
pvreohpiycllee,neseglleyctceodl ainndopolryoefthdpyrleefenereegnrlcyec,otlh.aTthpeertmeistttseudpbrsetpaanrcaetwiaonsotfetshteedhitgohdeesttesromliunbeletohre
`workablestock concentration,up solvents.
to
50mg/mL foraqueoussolventsand
500mg/mLfororganic:
PreliminaryToxicityTest
subs`tTahnepcreewloiumlidnbareytteosxtiecdit.yVteheisctlweaasndutseenddtooseeslteavbleilsshofthtehedtoesset-sruabnsgteaonvceerwewrheipclhattehde,otenset:
-
psellaetcetpievremdionsiem,awiltahgoavriernntihgehptrceusletnucreesaonfdaTbAs9eEn,cTeoA1f0A0r,ocTloAr1-Si3n5d,ucTeAdLrSa3tl7iavenrdS9W.PD2osuevlreAveolns
testedwere6.7, 10,33, 67, 100,333, 667, 1000,3333and5000pgperplate.
Mutagenicity Test
`Themutagenicitytest (initial andindependentrepeattests) wasusedtoevaluatethe m1u5t0a0geannidc5p0ot0e0ntuigalpoerftplhaettee)satsluobnsgtwaintche.apSpirxopdroisaetelveevehliscolfe atnedstpsousbisttiavnecceo(n1tr5o,l5s0w,er1e50p,l5a0t0e,d wAirtohcloTrA-iSn,ducTeAdIr0a0t,livTeArI9S.3S,AllTAdoIsSe37levaelnsdofWtPes2tuvsurbAstainnce,thveehpircelesecnocnetraolnsd aanbdsepnoscietiovef controlswereplatedin triplicate.
PlatingandScoringProcedures
origi"nTahleltyedsetscsryisbteedmbwyasAemxepsoseteadlt.o(1t9h7e5t)eastnsduubpsdtaantceedvbiyaMtahreoplnaatnedinAcmorepsora(1t9i8o3n)m.ethodology
(W/OV)n,twhaesdmaeylotefdiatnsdussuep,pmlienmiemnatletdwoiptahgLa-rh,icsotnidtianien,iDn-gb0i.o8ti%naangdarLt(rWy/pVt)opahnadn so0l.u5t%ionNtao Cal fwiintahl2co5nmceLntorfatwiaotnoerff5or0 ueaMchcac1h0.0mToLpoafgmairnniomtaulsetdopwaigtahr.SF9oorrtShheaprmempiarxatwiaosnsoufpmpeledmieanatnedd
~
BioReliance
-
Study No. AAS2XL.502001.BTL
2
`Company Sanitized. Does not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with anIndependent RepeatAssay
DuPont-8868
reagents,allreferencesto waterimplysterile,deionized waterproducedbytheMilli-QReagent `WaterSystem.Bottomagar was Vogel-Bounerminimalmedium E(VogelandBonner, 1956) containing 1.5%(W/V)agar. Nutrientbottomagarwas Vogel-Bonnerminimalmedium E containing1.5 % (W/V)agarandsupplemeatedwith2.5 %(W/V)Oxoid NutrientBrothNo. 2 (dry powder). Nutrient Broth was Vogel-Bonner sat solution supplemented with 2.5 % (W/V) `Oxoid NutrientBrothNo.2(drypowder).
Eachplate waslabeledwith acodesystemthatidentified thetestsubstance,testphase,dose.
love, testerstain, Procedures.
and
activation,
as
describedindetailin
BioReliance's
Standard
Operating
One-half 0.5)milterofS9or Shammix, 100kLoftester trainand25uLofvehicleor testsubstancedilutionwereaddedto2.0mL ofmoltenselectivetopagarat45:2C. After vortexing,the mixturewasoverlaidontothesurfaceof25mLofminimal bottomagar. When platingthepostivecontrols,thetestsubstancealiquotwasreplacedby aSOLaliquotof appropriatepositivecontrol. Aftertheoverlayhadsolidified,theplateswereinvertedand incubated.for approximately48 to 72hoursat 3742C. Platesthat werenot counted immediatelyfollowingtheincubation periodwerestoredat2-8Cuntilcolonycountingcould becondleusstchant10edayds).
=
`Thecondition ofthebacterialbackgroundlawnwasevaluatedforevidenceoftestsubstance:
toxicitybyusingadissectingmicroscope.Precipitatewasevaluated byvisualexamination
without magnification. Toxicityand degreeofprecipitationwerescoredrelativetothevehicle
control plateusingthecodesshown below.
[1] NSoiirgmhuayl [|DmiDsitisiinncgguuiirsshhoeeddbbcyyoaahnleoatloitchenyabylelthiannwingnof.t_he_mic_ro_co_lon_y _aw|n 2 ReduceYd |[aconmpdarpedossibtloythaesvleihgihctlienccornetraosleiplnatthe.esizeofthemicrocolonies| [Distinguishedby amarkedthinningofthemicrocolony Tawn| 3 | Moderted recosmpualretdingitnoatphevrehioclencoinontcrrouelasneinplcat theesidzeofthe microcolonis Distinguished byan extremethinningofthemicrocolonylawn 4 | ERxetdruemceeldy [rteosutlhteinvgeihincalneicnocnrteraosleiplnattheessuiczehotfhtathethmeicmrioccroolcoonlioesncyomlpaawnreids! vitsotihebunalideedeyeasisolatedcolonies.
[7 RE re greaterthanorequalto 90%oftheplate.
ep Precipitale_|tomicroscopic testsubstance precipitate.
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SBitouRdeylNioan.ceAAS2XL.5020BT0L1
13
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`Company Sanitized. Doo not contain TSCA CBI
aH.n2I5n1d7e1p:enBdaectnetrRiealpeRaetveArssseaMyutation Test with
DuPont8868
Non-Interferng|[DniaskteidnegyuiesbhuedtbaynypprreecciippiittaatteoepanrttihcelepsldaetechiadtbIystvhiseiabulteo(matthede Precipitate (ceo.lo,nlyecsosutnther3tapoarttinaclleslesosthnaanl1a0t%ewoiftthh3e0reevveerrttaanntics)o.lonycount ne |Dinsatkiendgeuyieshaenddbanyypprreecciippiittaatteeopanrttihcelepsldaetteecttheadtbiystvhiseiabulteotmoattehed] TPreciepitsrt e ||cmoolroentyhaconun3tpearrteicxlceeseodn1a0p%loatfetwhiethr3ev0erretvanetrtcaonlto)n.yThceosunetpl(at5e.s arecountedmanually.
contRreovlse,rtwenrtecocloounnitcesdfeoirtheargeinvteinreltyesbteyr saturtaoimnataenddaccotliovnaytcioounnctoenrdiotrioena,ireexlcyepbtyfhoarnpdosuintlievses the plate exhibitedtoxicity.
EvaluaoftReisuoltns
plFeowreraecchraepllicacteaupnldalatiranegr,ettphoeertmededa.nandstandarddeviationofthenumber of evertantper
-
meaFnorrotvheertteasnttsspubesrtpalncaetetoobfeaetvlaelausattoednepotsietsitveer,sittrmauinsotvcearusaemaidonsiem-uremlaotfedtiwncorienacsereianstihneg.
cpoosnicteinvteriafttihoensinocfrteeasstesiunbsmteaancner.eDvaerttaasnestaftohretpeestaekrstorftaihnesdToAsIeSr3esSpaonndseiTsAeLqSu3a7lweorfejgruedagteedr
tWhanZ3u,v0-rtAimweesrtehejmudegaend vpoeshiitcilveeciofntthoelivnaclrueea.sDeaitnamseeatn froervteesrttearnssiastitnhseTpAe9aEk,oTfAt1h0e0doasned
responseis equal o o graterthan2.0-imesthemeanvehiclecontrolvalve.
Criftor aeVarlidiTeast
Salm"oTnheelfloalltoewsitengscrriitnecruilatmuuresstmbuesmtdeetmfonosrttrhaetemtuhteagperneisceintcyeteosftthteodbeeepcornosuigdhemruetdavtailoind.(A7al)l
tahnedptrheesdeenlcteioofntinhtehpeKuMvrIB0g]epnlea.sCmuildtRu-refsactofotre.stAelr sltWraPi2nsuvTrAASBcualntdurTeAsmIu00smtudesmtodnesmtornasttertathee
dsponetanleionuetshrte eivAoegeirnnnet.thAaelvnechutilctsluerecsonmtursoltsdaesmonfsotllroawtset(hineclcuhsairvaec)t:erTiAs9t8i,cm1e0a-n50n;uTmAbeIr00o,f
n8u0m-b2e4r0s;oTfAbIaSct3eSr,ia5a-r4e5p;leTAdI,StYe7st,er3s-t2r1a;incWuPlt2uurerAti,ter1sm0-u6s0t.bTeogreenastuerrethtahnatorapepqruoaprliaftoe
t0h3ex1n0u"mcbeelrlosf/meLv.eTrthaenmiesoavneorftcheacmhepaonsiviavleuecoonfttrhoelrmeusspetcetxihviebivteahticlleeacsotntar3o.l0.-Afolmdiinncirmeausme ionf
thonreeoenrobno-ttohxoifctdhosfeollelvoewlisnigsrcreiqtueirriaedarteo emveatlua(1e)eAst>d5a0ta%.rAeddouscteiloenvielitshceomnseiadenrendumtboexircoiff
-
BStiuodRyelNioa.nceAAS2XL.502001BTL
14
Company Sanizad. Does notcontain TSCA G81
H-25171: BacterialReverseMutationTestwith anIndependent RepeatAssay
DuPont-8868
revertants perplateascompared 1themeanvehiclecontrolvalue. Thisreductionmustbe ianctchoempbaancikegdrbouynadnlaabwrnu(pbtdaocskeg-rdoeupnedncdoednetdorfo3p,i4n othre5r)e.veArctaonptycooufntth.e(2H)isAtomroidcearlaNteegartediuvcetainodn PositiveControl ValuesisincludedinAppendixA.
Archives
All raw data,
andallreportswillbemaintained accordingtoStandard
Operating
theBioRelianceRAQAunitheadquarteredat:BioReliance,
1le4a9s2tt0hBrreoesycehaarrstafter whichtimeMthDe 2S0p8o5ns0o.Pr wriltlhibseScOoPn.tapcatpeedrfroercora ddeswciislilobnearsettaoitnehdeffoirnaalt
cdiosppioesditainodntofhtehceompaytewriilallsb.eArleltasitnueddyimanttehrieaBlisorReetluimaendcteoatrhcehiSvpeosnfsoorr oarmdienstirmouymeod wfi1ll0fyierastrbse.:
`Unuseddosingsolutionsweredisposedof followingadministrationtothetestsystem andall
residualtestsubstancewillbedisposedoffollowingfinalizationofthe report.
Deviations Noknowndeviations fromthe protocolorassay-method SOPsoccurredduringtheconduct
ofthisstudy.
BioReliance
--
Study No. AAS2XL.502001.BTL
15
CompanySanitized. Doesnot contain TSCA CBI
aH-n25I1nd7e1p:enBdaectnetrRieaplReaetvAerssseayMutationTest with
DuPont-8868
RESANU DDILSCUTSSISON
Solubility Test
targeTtecterlalhsyadrnodfsuorlaunb(iTliHtFyo)wfatshseetleescttseudbassttahneces,olTvheeattoefstcshuobisctebanacseedwoansscoolmupbalteibbiultitcylwoiutdhytihne tetrahydrofuran(THE) at amaximumconcentrationofapproximately200mg/mL.
PreliminaryToxicity Test
wer`eTgheenreersautletdsoifntEhxepperreilmiemnitnAarLytIonxtihceipyrteelsitmairneaprryetosxeinctietdytienstT,atbhleesma1xtihrmouumgdho4s.eTthesetseeddwataas
5000 perplat;thisdosewasachievedusing aconcentrationof200mg/mLand a25 uL
palnadticnlgeaalrisqouloutt.iTohnfetreosmts0u.b2s7ttanoc2e7wamsg/amwLor.kDaobsleesloelvuetlisotnefsrtoemdw4e0reto 62.07,0m10g,/3m3L,6a7n,d 1a0s0o,l3u3bl3e,.
6N6o7a,p1p0r0e0c,ia3b3l3e3toaxnidci5 tywa0 usgopb0 esreprlv0 aetde.,DPrueecitpoittaetsetweasstorbaisnecruvletdubreegcionnntianmginaatti1o0n0,tuegspteerrsptlrataei.n
--
`toTxAiLci0t0y.inBtasheedpornestehnecfeinadnidngasbosfetnhceetooxficit9yatcetsitv,atthieomnawxaismounmldyeovsaelpulaatteeddifnotrphreemcuitpaigteantiecaintdy
testwas5000 ugperplat.
Mutagenicity Test
`Theresultsofthemutagenicitytestarepresentedin Tables $through24andsummarizedin `Ta2b5laned2s6.ThesedataweregeneratedinExperimentsBIand B2.Doselevelstested were 15,50,150, 500, 1500andS000 ugperplate. Prewcasiobsperivedtbegainntingeat 500 ugperplate. Noappreciabletoxicity wasobserved.Forsubmissionto Japaneseregulatory agencies,pertinentadditionalinformationisincludedinAppendixC.
obseIrnveEdxwpietrhiamneynotftBIhet(eIsntiteirasltrMauitnasigenneiictihteyrthTeesptr)e,sennocepoosraibtisveencmeuotfagSe9naicctirveastipoonn.ses were
obsIernvEexdpweitrhiamneynotfBt2he(tIensdteepestnadiennsitRneeiptehaertthTeepstr)e,snenocpeoosriatbisveenmcuetoafgeSn9aicctrievastpioonns.es were
CONCLUSION
BactAelrilaclriRteevreiarfsoerMuatvaatliiodnsTteusdtywwietrheamneIntdaespdeensdcernitbReedpienatthAespsraotyoicnodli.cTatheethraetsu,ultnsdoefrtthhee
cpornedsietnicoenosroafbtsheinscsetoufdAy,rHo-2c5l17o1rd-idirnnaottdlciuavecurse$e9d.apositivematagenicresponsein citherthe
-~
`BioReliance
---
StudyNo. AAS2XL.502001.BTL
16
Company Sanitized. Does not contain TSCA CBI
aHn-2I5n1d7e1p:enBdaecntetrRiealpReaetveArssseaMyutationTestwith
DuPont-8868
REFERENCES Ames, BN., J. McCannandE. Yamasaki (1975)MethodsforDetectingCarcinogensand
Mutagenswith the SalmonelMiclroasom/eMMutaagenmicimtyaTeslt, iMutaatinon Research, 31:347-364.
Green, MH.L.andW.J. Muriel (1976) Mutagentestingusingtrp reversioninEscherichiacoli, Mutation Research 38:3-32.
InternationalConferenceon Harmonisation(ICH)ofTechnicalRequirementsforRegistration ofPharmaceuticalsfor Human Use.GuidanceonSpecificAspectsofRegulatory GenotoxiciyTestsfor Pharmaceutical. S2Adocumentrecommendedforadoptionatstep 4 oftheICHprocoenJsulsy 19, 1995.Federal Register 61:18198-18202,April24, 199.
InteronfatPihoanramlaCcoenufteircaelnscefoornHHuarmmaonnisUastei.onGe(nIoClHox)iociftTye:chAniSctaalnRdaerqduiBraetmteenrtysffoorrGReengoisttorraitciiotny TestingofPharmaceuticals. S2B documentrecommendedforadoptionatstep 4oftheICH proconeJsulsy 16, 1997. FederalRegister62:16026-16030,November21, 1997.
Maron, DM. and B.N. Ames (1983) Revised Methods for the Salmonella Mutagenicity Test,
-
MutationResearch, 113:173-215.
`OECD Guideline 471 (Genetic Toxicology: Bacterial Reverse Mutation Test), Ninth Addendum 0the OECDGuidelinesfortheTesting ofChemicals,publishedbyOECD,Paris, February 1998.
Voge`lPr,oHpeJr.tiaens,dJD. .BiMo.l.BCohnenme,r2(1189:5967)-A1c0e6t.ylomithinase ofE. col: PartialPurificationand Some
BioReliance
-
Study No. AAS2XL.5020B0T1L
1"
Company Sanitized. Does not contain TSCA CBI
aHo-25171: BactetrRiepeaat l ReveArsssaeyMutation Test with
DuPont-8868
PBraecltienriinaalryMuTtoaxtiicointyTTeesstt
Table 1
TSetsut.dysNuubmsbtearnce Id:: AAH-S2X5L1.75102001.B7L
stExrpaeirniment No. :: TALass
VDeahtieclPelated :: 2t0etDreachya20r0o1turan
Plating Aliquot : 25 yi
TCeosntceSnutrbasttiaonnce RevWeirtthan5t9s ActBiavcaktgiroonund
ug per plate per plate Lawn
Venicle
2
1
WReivtehrotuatnt5s9 AcBtaicvkagtrioounnd
per plate Lawn.
2
1
61.07
32
11
119
11
33
234
i1
116
11
130303
229
wmee
133
wwee
-
106070
226
wmee
i1d
nwee
35030303
230
wnee
112
wwee
BackgroIu-NnodrmLaalw;n C2o:d5e1ightly reduced; 3=Moderately reduced
N4P--ENxotnr-eImnetleyrfreerduicnegd;preSceiApbisteantte;; 6=IOpbssIcnutreerdfebryinpgrepcriepciitpiattaete
BioReliance
~
Study No. AAS2XL.502001.BTL
18
Gompany Sanitized. Does not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with
anindepen~~dentRepeatADusPons8ia63y
PBraecltiemriinaalryMuTtoaxtiicointyTeTsetst
able 2
TeSBstitupdeySruimbeusntmtabnecNroe. 14:|:: HAAL-R2S5I1X7L1.502001.BTL
StDartaeinPlated i: m20usDeac 2001
VePlhaiticngle iquoe ii c25etrahyaroturan
TCeosntcensturbasttainocne RevWeirtthan5t9s ActBievcaktgiroonusd RWeivtehrotuatnts59 AcBtaicvkagtrioounnd
0 por plate per plate Lawn per plate Lown
Venice
15
x
2
T
61.7 a3
110
13
n
1
11
11
is
1
10a0
15
1me
i1s
1we
-
3se3
112
mmee
br1}
wwee
To00
1s
me
c
50330303
10a
mmee
2102
wmee
BackgrSo-uNnodrnLaelv,a C3o-dSelightly reduced; 3=koderately reduced
NApo-ENxotnr-emIenltyerfreedruicnegd;preSc-iApbistanatt;e; 6I=7O-bIanctuerrefdebryinpgrepcriepciitpaittaete
Clcontaninated
BioReliance
-
Study No. AA52XL.502001.BTL
19
Company Sanitized. DoesnotconTtSaCAiCBnI
'H-25171: Bacterial Reverse MutationTest with
anlndepen~~dentRepeatADuPsonst88s68y
PrBeacltiemriinaalryMuTtoaxtiicointyTeTsetst
Table 3
TeSsttudsyNubusmtbaencre 1a:+ AH-A2S5I1X7L1.502001.BTL StErxpaeirniment No. : mALasa VDeantieclPelated:i 2t0etDreachy2d0r0o1furan
TCoensctensturbasttainocne ReveWritthant59s AcBtaicvkagtrioounnd RWeivtehrotuatnts59 AcBtaicvkagtrioounnd
_bgpeVrepnilcaete perplate LaT wn perpla7te L1ew
p5H.7 3
107
11
2
1
1s
13
7
1
10@0 3
15
w1 e
10
me
55
1we
5
we
-
Tsoo00 3933
04
wmee
5
me
12o
wmee
7
me
5000
10
we
:
wp
BackgroIuenNodrmLaalv;a C2o-dSe1ightly Extremely reduced;
rSeedAubcaeedn;t;
Is6M-oodbescruarteedlybyrepdruececdipitate
NeeNon- Interfering precipitate; IPeIntorfering precipitate
BioReliance
--_
Study No. AAS52XL.502001.BTL
20
Company Sanitized. Does not contain TSCA CBI
H-25171:Bacterial Reverse MutationTestwith an Independent Repeat Assay
DuPont-8868
PBraecltiemriinaalryMTuotxaitciointyTeTsetst Table
ToSottudySuNbusmtbaenrce Ia:i AHN-S225X17L1.502001.BTL
StExrpaeirniment No. : WALe2 uve
VDeahtieclPelated:+ t20etrDeacny2a0r0o1furan
Plating Aliquot : 25 ui
TCeosntceSnutrbasttiaonnce RevWeirtthant9s ActBiavcaktgiroonund
ug per plate per plate Lawn
Vehicle
2
1
WReivtehrotuatnt5s9 AcBtaicvkagtrioounnd
per plate Lawn
18
1
160.7
2225
11
2124
11
3&3
1250
11
1175
11
130303
1159
wwee
231
wwee
-
1056010
225
mwee
3n1
wwee
50330303
1286
mmee
1u8
wnee
BackgroTuenNdormLaalw;n C2o4dSelightly reduced; 3=Moderately reduced
N4PcEexotnr-eImnetleyrfreerduicnegd;preSceiApbisteantte;; 6-ICPbssIcnutreerdferbiynpgrepcriepciitpaittaete
`BioReliance
~~
Study No. AAS2XL.502001.BTL
21
Company Sanitized. Doesnot conTtSaCAiCBnI
H-25171: BacterialReverseMutation Testwith an Independent Repeat Assay
DuPont8868
Bacterial Mutation Test
Table 5
TSotoutdySNuubmsbtearnce Id:: HA-A2S5Z1X7L1.S02001.BTL Experiment No : BL
sLtirvaerinMicrosomes :+ Nmoonse
DCaetlelsPSleaetedded :: 42.J7aXn 2100%02
VPelhaitcilneg Aliquot : t25eturiahydrofuran
Cugoncpeerntprlaattieon NPulmabteer pReevreprtlaanttes BacCkogdreound RAevveerratganets SDteavnidaatridon
Vehicle
0021
115 11
5
20 1
16
3
15
o0a2
210 11
0
11
15
5
s0
002
118 11
03
12 1
1
3
150
002
116 11
03
10 1
1B
3
500
002
1153 mwee
03
10 we
5
3
1500
0a2
11 mmep
0
1 we
15
2
5000
0a2
1102 w1ep
03
12 ie
un
1
Positive Control 20-1nitrofluor21e7ne 1.0 ug1 per plate
o0a2 128302 11
am
2s
Backgro1ueNnodrnLaalw;n C2oed5e1ightly reduced; JsNoderately reduced
N4p-aENxotnr-eImnetleyrfreerdiucnegd;preSceiApbistenatt;e; 6=IOPbssIcnutreerdferbiyngprperceicpiiptiattaete
BioReliance
~
Study No. AAS2XL.502001.BTL
2
Company Sanitized. Does not contain TSCA CBI
H-25171: Bacterial Reverse MutationTestwith
a
t Assay
DuPont-8868
Bacterial Mutation Test
Table
TSetsutdySNuubmsbtearnce Td:: AH-A2S5Z1K7L1.502001.BTL Experiment No : Bl
StLirvaeirnMicrosomes : RTaatssliver 59
DCoaltleaPSleaetedded :: 42.J7anx 210002
VPleahtiincgle Aliquot :+ 2t5etruaihydrofuran
Concenrtrapltaitoen NPulmabteer Revperertplaanttes BackCgodreound RAevveerratgaents DSteavnidaatridon
Vehicle 0021
3230 11
03
21
2
7
15
0012
2266 11
0
21
25
2
50
0012
12s7 11
03
nw 1
20
s
150
002
2328 11
0
=n 1
2
6
00
o012
225 wwee
0
20 we
2
3
1500
o012
3358 mmee
0
25 we
3
7
5000
0021
125 11pp
0
20 1p
2
4
Positive Control o2-1aminosnthr48a0cens 1.01ug per plate
002
83338 1a
a3
oa
BackgroIueNnodrnLaalw;n C2oedSelightly reduced; JaModerately reduced
N3e-wENxotnr-eImnetleyrfreedruicnegd;preSceiApbistenatt;e; GeIOPbssIcnutreerdferbiyngprperceicpiiptiattaete
BioReliance
~
Study No. AAS2XL.5020B0T1L
2
Company Sanitized. Does not contain TSCA CBI
'H-25171: Bacterial Reverse Mutation Test with an Independent Repeat Assay
'DuPont-8868
Bacterial Mutation Test
Table 7
TSetsutdySNuubmsbtearnce 1a:: AH-A2S5I1X7L1.502001.BTL Experiment No : BL
StLirvaeirnMicrosomes : N1o0ne0
DCealtlesPlSaeteedded :: 41.1JaXn 210002
VPelhaiticnlge Aliquot:+ 2t5etrpal,hydcoturan
C4gonpcoerntprlaattieon NPulmabteer pReerveprtlaanttes BackCgodreound RAevveerratgaents DSetvainadtairdon
Vehicle 0012 129203 11
03 182 1
198
2
15
o0n2 116683 11
03 179 1
170
5
s0
0012 116861 11
0 168 1
182
fg
150
002
115799 11
03
150 1
176
2
500
0021
1160s wwee
03 c
103
`
1500
0012 127092 wwee
03
103 we
175
30
5000
002 126036 wwee
03
152 we
187
2
Positive Control s0o1dium azide721.0 ug pe1r plate
0023 a6s1i8 13
714
a2
Backgr1o-uNnodrmLaalw;n C2o-dSelightly reduced; 3=Moderately reduced
N4p-eBtxitorne-mTenltyerrfeedruicnegd;preSc-iApbistenatt;e; GI=PO=bIsnctuererdferbiyngprperceicpiiptiattaete
CContaminated
---
BioReliance
Study No. AAS2XL.5020B0T1L
%
`Company Sanitized. Does not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with
an
t
DuPont8868
Bacterial Mutation Test
Table
TSetsutdysNuubmsbteanrce Id: AH-A2S521K7L1.S02001.BTL Experiment No : BL
stLirvaeirnMicrosomes :+ R1a0t 0liver 9
DCoaltleePlSaetedeedd :: 41.1JaXn 210002
VPelhaitcilneg Aliquot:: t25etrahydrofuran
1Cgoncpeenrtprlataitoen NPulmabteer pReerveprltaatnets BackCgodraound __ARveevreratgaents DSetvainadtairodn
Vehicle 0012 161531 11
03
208 1
3
2
15
0012
2102 11
0
159 1
180
2
s0
0012 119700 11
-
03
78 1
175
10
150
0012 19m8 11
0
105 1
185
1
500
o012 1m95 wmee
03 165 me
1m
16
1500
a02 12617 wwee
03
Fd we
193
19
5000
a02 119853 mmee
0
226
we
200
22
Positive Control 20-1aminoanthr87a7cene 1.01ug per plate
0032 872520 11
816
8
Backgro1u-nNodrmLaalw;n C2o=dSelightly reduced; JsModerately reduced
Nie-sBNxotnr-eImnetleyrfreerdiucnegd;preSc=iApbisteantt;e; G=IOPb-sIcnutreerdfebryinpgrperceicpiiptiattaete
--
SBtiuodRyelNioa.nceAAS2XL.502001.BTL
25
CompanySanitized. Doesnot contain TSCA C81
'H-25171: Bacterial ReverseMutation Test with
om
tRepeatAssay
DuPont-8868
Bacterial Mutation Test
Table 9
TSetsutd.ySNuubmsbtearnce Id:: HA-A2S5Z1X7L1.502001.BTL
stLirvaeirnMicrosomes : NTAoLnSe3s
Vehicle
+ tetrahydrofuran
CExoplelreimSeenetdeNdo :: B2L.6 X 10' Date Plated : 4 Jan 2002
Cugoncpeerntrapltaitoen vehicle 15
NPulmabteer Revertlaanttes BackCgordeound
0012
115 11
03
11
0012
1156 11
03
15 1
RAevveerratgaents SDteavnidaatridon
15
`
1
1
s0
o0a2
1I2 11
-~
03
11
1
3
150
o021
125
11
03
15 1
1
s
500
0021
1232 mwee
0
15 we
w
s
1500
002
11s2
wwee
03
5 we
2
`
5000
o0a2
1128 11pp
0
1 we
16
Positive Control s0o1dium azide021.0 ug pe1r plate
0023
4pi7t6s 11
a1
Backgro-uNnodrnLaalw;n C2o-dSe1ightly reduced: 3eModerately reduced
N4PneBNxotnr-emIenltyerrfeedruicnegd;preSc=iApbistenatt;e; G=IOpbssIcnutreerdfebryinpgrepcriepciitpaitteate
BioReliance
-
Study No. AAS2XL502001BTL
2
Company Sanitized. Doesnot conTtSaCAiCBnI
'H-25171:Bacterial ReverseMutationTestwith
am
t
DuPont-8868
Bacterial Mutation Test
Table 10
TSetsutdySNuubmsbtearnce Td:|: HA-A2S521X7L1.502001.BTL Experiment No : Bl
stLirvaeirnMicrosomes : TRAaItS3lSiver $3
CDoaltleaPSleaetedded :: 24.6JanX 120002
Vehicle
: tetrahydrofuran
_CPolnacteinntrgaAtliioqnuotP:la2teSpLRevertants Background Average Standard
ug per plate Number per plate Code _Revertants Deviation
Vehicle 0012
1251 11
0
25 1
20
5
15
o012
226 11
0
21
2
3
s0
0o21
1162 11
-~
03
16 1
15
2
150
a02
1200
11
03
11
15
s
500
0021
210 mwee
0
15 we
16
3
1500
0012
11 wmee
03
5 we
12
3
5000
o021
213 wwee
03
2 we
1
s
Positive Control 2-01aminoanthr17a3cene 1.01ug per plate
0023 121686 11
09
2
BackgroeuNnodrmLaalw;n C2o-dSelightly reduced; 3=Moderately reduced
N4Pe=ENxotnr-eImnetleyrfreerdiucnegd;preSc=iApbisteantt;e; 6=IOPbsaIcnutreerdfebryinpgrepcriepciitpaitteate
BioReliance
-
Study No. AAS2XL.5020B0TL1.
27
`Company Sanitized. Does not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with anlndependentRepeatDAuPsous8Ss6sy
Bacterial Mutation Test
able 11
mSetsutdySNuubmsbtaenrc|e Id:i AB-A2S5I1X7L1.S02001.BFL SeLixvaeirnMicrosomes :$ mNoanse y
CBoplesrimSeenetdeNdo Date Placed
:: :
B1L.3 X 4 Jen
120002
VPelnaitcilneg liquor : t25etruiahydrofuran
CSoonpceerntrpalattieon NPulmabteer pReerverptlaanttes BackCgordeound _ARevveerratgaents SDteavnidaatridon
Venscle 00 o
37 1I a1
s
2
1s
0o
s8 1i
1I
s
.
-
s0
00 0
3a 13 31
s
5
150
o0 o
n5 11 c1
7
3
s00
0a 03
.5 mwee s we
s
1
1500
0o2n 0
3s minee s we
.
1
s000
ooo
[H
55 Imee 3 me
El
3
Positive Control 90-aminoacrid50ine 75 pg2per plate
0o Tr Go
35si2s 11
s0 16
eT meme
NeTpREooxNrtomrnae-lmT;enity2r.fr8ee1rdiuigcnhegtdl;yprreSecediAupbcisetedna;tt;e;ISseTMOoPbdIsencrtuaertreefdleyrbiyrnegpdrupecrecdeipciitpaitteate
yy
BioReliance
-
Study No. AAS2XL.502B0T0L1
28
`Company Sanitized. Does not contain TSCA CBI
'H-25171: Bacterial Reverse MutationTestwith nlndependentRepeatADusPosn8s 6sy
Bacterial Mutation Test
able 12
TSetsutdysNuubmsbteanrce Strain
1d:+ MH-S2I5X1L7.1502001.BTL {mas
Liver Microsomes : Rat liver $9
EDCxeapltelersiPmSleeanetdceeNdod:::
B1l.3 X Jan
210003
VePlhaciicngle Aliquot : C2e5trual.hyarofuran
CSoonpceerntrplaattieo.n NFulmabteer Rpeevrerptlaanttes PacCkogdreousd _ARevveerratgaemts SDteavnidaatridon
Venicle o0n 0
13s 1i 81
10
3
1s
oao o
1535 113
.
s
-
so
00 0
107` 11i
.
2
130
oa
52 11
0 10 1
5
2
s00
o0 os
.8 wnee 5 me
7
3
1500
00 o
sa wwee : ne
`
2
s000
o0r 0
55 1impe 5 we
7
2
Positive
Control
20-aminoanthra5c6ene 0 103
1.0 2 ug 1
per
place
0 se 1
5
10
BackgroTiuoinNEodxrinLraaelvn;aciCy3o0dr8ee1diugchetdl)y rSe-dAubcaeedn;t; Js6M=oOdbesrcautreeldybyrepdruecceidpitate
NpeNon- Interfering precipitate; IPsInterferineg prmeciptitataermrest
`BioReliance
--_
Study No. AAS2XL.50200B1T.L
29
Company Sanitized. Does notcontain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with an IndependentRepeatAssay
DuPont-8868
Bacterial Mutation Test
Table 13
TSetsutdySNuubmsbtearnce Id:: AHA-S2Z51K7L1.502001.BTL Experiment No : BL
SLtirvaeirnMicrosomes : Neozneuvin
DCoaltlesPlSaeetdeedd :: 24.5JaXn 120002
VePlhaitcilneg Aliquot :: t25etruathydrofuran
C0onpceerntrpalattieon PNulmabteer Rpeevreprltaatnets BacCkogdreound _ARevveerratgaents SDteavnidaatridon
Vehicle 0o21
112 11
0
15
1
u
2
1s
0a2
11 11
0
13 1
1
2
50
0021
1162 11
---
03
11
1B
3
150
0021
116 11
0
10 1
3
3
s00
o012
1u0n
wwee
0
Fd we
3
4
1500
002
118 wwee
0
12 we
1
3
5000
o0a2
110 1irpe
03
8 1p
10
2
Positive Control ome1thyl metha8n5esulfonat1e 1000 ug per plate
0023
992 11
0
a
BackgroLueNnodrnLaalw;n C2oedSelightly reduced; JsModerately reduced
Ne4eeNBoxntr-mInetleyrfreerdiucnegd;preSceiApbisteantt;e; 6=IOPb=aIcnutreerdfebryinpgrepcriepciitpaitteate
BioReliance
-
Study No. AAS2XL.502001.BTL
30
Company Sanitized. Does not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with
an In
Assay
DuPont-8868
Bacterial Mutation Test
Table 1
TSetsutdySNuubmsbtearnc~e Id:: HA-A2S5Z1X7L1.502001.BTL Experiment No : BL
stLrivaeirn Microsomes :+ wRea2t luivviear S9
CDealtlesPSleaetdeded :: 24.5JanX 210002
VPleahtiicnlge Aliquot :: t25etruabhydrofuran
Concentration : plate
NPulmabteer Revperertpalanttes BacCkogdroeund
RAevveerratgaemts SDetvainadtairodn
Vehicle
0012
1169 11
03
12 1
16
s
1s
002
1186 11
0
u1
15
50
o012
126 11
-
03
11
v
`
150
0021
1160 11
0
13 1
13
3
500
o0a2
110 wmee
03
u we
12
2
1500
0012
1165 wwee
03
1 we
15
2
5000
0012
1I amee
03
5 we
3
s
Positive Control 2-o1aminoanthr56a5cens 10 u1g per plate
0032
87489 11
766
191
BackgroNuonrdmaLalw;n C2o-d5elightly reduced; d=Moderately reduced
4NePEsxocnr-eImnetleyrfreerdiucnegd;preSceiApbisteantte;; GeIOpb=sIcnutreerdfebryinpgrepcriepciitpaitteate
BioReliance
---
Study No. AAS2XL.5020B0T1L
31
`Company Sanitized. Doesnot contain TSCA CBI
'H-25171: BacterialReverse Mutation Test with. an IndependentRepeatAssay
DuPont.8868.
Bacterial Mutation Test
Table 15
TSetsutdySNuubmsbteanrce Id:: HA-A2S5Z1X7L1.502001.BTL
Strain
+ mos
CEexlplesrimSeenetdeNdo :: B12.0 X 10'
VLeihviecrleMicrosomes +: Ntoenterahydrofuran Date Plated : 15 Jan 2002
Plating Aliquot : 25 wb Concentration Plate Revertants Background Average
Standard
us late Number per plate Code Revertants Deviation
Vehicle 0o21
237 11
0
21
2
s
15
0012
216 11
03
29 1
zn
7
-
s0
0021
126 11
03
21
2
7
150
0o21
229 11
0
21
2
1
500
0021
11s3 wwee
03
1 we
16
2
1500
o02
232 1r1e2
0
13 re
2
10
5000
0012
331 1rpe
03
1 re
2
12
Positive Contxol 2o-1nitrofluor1e58ns 1.0 pg1 per plate
002
m58 11
12
38
BackgroIuenNdormLaalv;n C2o5d1eightly reduced; 3=Moderately reduced
4NepE-xotnr-eImnetleyrfreerduicnegd;preSc=iApbisteantte;; =IOpbssIcnutreerdferbiyngprperceicpiiptiattaete
BioReliance
-
Study No. AAS2XL.502001 BTL
2
Company Sanitized. Does not contain TSCA CBI
'H-25171:BacterialReverse MutationTestwith
an
Assay
DuPont.8868.
Bacterial Mutation Test
Table 16
TSetsutdySNuubmsbtearnce Id: AHA-S2Z5X17L1.502001.BTL
strain
mss
ECxeplelsrimSeenetdeNdo :: B12.0 X 10
VLeihvierclMeicrosomes :+ tRaettralhiyvderroSf9uran Date Plated : 15 Jan 2002
Plating Aliquot : 25 ub C4goncpeenrtrapltaitoen NPluamtbeer pReervertplaanttes
BacCkogdreound
__ARveevreartgaents
SDetvainadtairodn
Vehicle 0021
1362 11
o
26 1
25
15
0o21
2268
11
0
31 1
28
3
50
0021
3@s 11
-
0
18 1
3
15
150
0021
3a5 11
03
a1
a
7
500
o012
3385 wwee
0
20 we
n
10
1500
002
a3 mwep
03
2 we
3s
5
5000
0012
3270 1ipe
03
32
re
a
7
Positive Control o2-1aminanthr6a3c1ens 1.01ug per plate
0032 875182 11
734
mn
BackgroIu-nNodrmLaalw;n C2o-dSelightly 4cExtremely reduced;
rSe=dAubcseedn;t;
d=MeoOdbesrcautreeldybyrepdruecceidpitate
NpNon- Interfering precipitate; IPInterfering precipitate
BioReliance
~
Study No. AAS2XL.502001BTL
3
`Company Sanitized. Does not contain TSCA CBI
H-a25171: BactetrialRevAesrssaeMyutationTestwith
DuPont-8868
Bacterial Mutation Test
Table 17
TSetsutdysNuubmsbteanrce 1d:: AH-A2S5I1X7L1.502001.3TL
strain
+ TAL00
CEexlplesrimSeenetdeNdo :: B02.9 X 10
VLeihviecrleMicrosomes : Ntoenterahydrofuran Date Plated : 15 Jan 2002
Plating Aliquot : 25 Concentration Plate
uL Revertants
plate Number per plate
BacCkogdreound
RAevveerratgaents
DSteavnidaatridon
Vehicle 002
114250 11
03 100 1
122
20
15
o012 151500 11
0
78 1
161
1s
_
50
0012 156 1 0 178 1
167
16
150
002 1286 11
0
167 1
107
2
500
o0r2 117287 wwee 0 156 we
15
26
1500
002
1m512 wmee
0
52 we
138
2
5000
0021 110306 aape 0c
120
2
Positive Control so01dium azide3a1.0 bg pe1r plate
w 02 d 364 a 1 66 20
BackgreouNnodrnLaalw,n C2o-dse1ightly oixtremely reduced;
SreeAdbusceendt;; 3=GM=oOdbescruarteedlybyrepdruecceidpitate
Ncpaectoonnt-anIinntaetrefedring precipitate; Ip=Interfering precipitate
-
BioReliance Study No. AAS2XL.502001.BTL
4
Company Sanitized. Does not contain TSCA CBI
H-25171:BacterialReverse MutationTestwith
mw
Assay
DuPont-8868
Bacterial Mutation Test
Table 18
TSotsutdySNuubmsbtearnce Id: AH-A2S5Z1X7L1.502001.BTL Experiment No : B2
stLirvaeirnMicrosomes +: 1Ra0t 0liver $9
CDoaltleePlSaetedeedd :: 01.59JXan102'002
vehicle
: tetrahydrofuran
MCaontcienntgraAtliioqnuotP:la2tespRnevertants Background Average Standard
bg per plate umber per plate Code Revertants Deviation
Vehicle 0012
117512 11
0 193 1
m2
2
15
0012
215000 11
0
156
1
18
26
s0
002 116092 11
---
03
181 1
m
7
150
o0a2 116550 11
03 161 1
159
5
500
0012 11628 mwee
03
de we
151
1
1500
0012
1m511 1rpe
3
116
ire
46
28
5000
002 116897 1rpe
03 156 112
m
16
Positive Control 2-01aminoanthr83a7cens 1.02ug per plate
0032 s7e1a0 11
738
8
Backgro1uenNodrmLaalw;n C2o=dSelightly reduced; 3=Moderately reduced
4Np=aEtxitorn-eImnetleyrfreerdiucnegd;preSc=iApbisteantte;; 6=IOPb=sIcnutreerdfebryinpgrperceicpiiptiattaete
BioReliance
--
Study No. AAS2XL.502001.BTL
3s
Company Sanitized. Does not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with
a
Assay
DuPont-8868
Bacterial Mutation Test
Table 13
TSetsutdysNubustmabncee r1d:: HA-A2S5I1X7L1.502001.BTL Experiment No : B2
sLtirvaeirnMicrosomes : Nmao1nse3s
CDaetlelaPSleaetedded :: 105.9JaXn102002
VPelhaicicnlge Aliquot: 2t5etrahydrofuran
Cugoncepenrtrapltaitoen NPulmabteer Repverertplaanttes BacCkogdreound _RAevveerratgaents DSteavnidaatridon
Venicle 0o1z
38 11
03
12 1
un
3
15
o0n2 1160 11
03
11
2
3
s0
002
1un3 11
~
0
51
n
2
150
002 1u6n 11
0
11
3
3
500
o02
55 rzee
0
1 ne
5
2
1500
0o2
1128 1ape
03
un ae
fn
4
5000
001
Fe1]3 uaree
03
16 re
1
2
Positive Control so01diun azi1de791.0 ug pe1r plate
0023 128156 1i
193
20
Backgroiu-nNodrmLaalw;n 2Co=dSelightly reduced; J=Moderately reduced
N4P--Btxotnr-eInnetleyrfreerduicnegd;preSc-iApbisteantte;; GI=PO-bIscnutreerdfebryinpgrperceicpiiptiattaete
--~
BioReliance
Study No. AAS2XL.50200B1T.L
36
Company Sanitized. Doesnotcontain TSCA CBI
H-25171: Bacterial ReverseMutation Test with an Independent Repeat Assay
DuPont-8868
Bacterial Mutation Test Table 20
TSetsutdySuNbumsbtearnce 14:: HM-A2S5I1X7L1.SO0001.TL Beperiment No : 52
sVtLeirnvaieicrineMicrosomes : TRtaaeittsrialshiyvedrrof$u9zan cDaecleiaPSleactedded :: 01.35Jaxn102002
Plating Aliquot : 35 ub
Concenrtrapltaitoen NPulmabteer Revperertplaanttes BacCkogdreound RAevveerrtaagnets DSetvainadtairdon
Vehicle 02013
31s 1ii
un
3
is
0001 x10s 1i
2
`
s0
o00r23 11i20 3ii
0
s
-
150
oo012 11333 1ii
13
s00
0001 ii1so mmveee
1
`
1500
o00r23 1n00 1iimr1ee
2
`
s000
001
1o airree
0
1 ire
13
.
Positive Control 2o-02raminoant1hri0a6cene 1.0viug per plate
Background Lawn Code0.
83
112
2
NIip-EeNxlotorrnne-anlI;onltye3r-fr5eleridigunhcgteldpyrerSc-eiAdpbuiscteenadt;t;e;3s-MIOoPbdsaeIcrnuatrteeerdlfyeLryrienpdgrupecrecedicpiiptiattaete
NeToececiouvnetanduealitqouoptrocoefdutreaslterersrtorraiinn which tho plate did not
--
SBtiuodRyelNioa.nceAAS2XL.5020B0T1L
37
Company Sanitized. Does not contain TSCA CBI
H-25171: BacterialReverseMutation Testwith
an
t Assay
DuPont8868
Bacterial Mutation Test
Table 21
TSetsutdySuNbusmtbaenrce Id: HA-A2S5Z1K7L1.502001.BTL Experiment No : BZ
stLirvaeirnMicrosomes : NTo1n5e37
DCealtlesPlSaeteedded :: 105.5JaXn102002
VPleahtiincgle Aliquot :: 2t5etrahydrofuran
bCgoncenpertrapltaitoen NPulmabteer Revperertplaanttes BackCgodreound RAevveerratgaents DSetvainadtairodn
Vehicle 000312
524 111
`
2
1s
002
56 11
0
`1
s
1
50 0002 77s 111
5
2
150
0a2
52 11
0
51
2
s00 a002 74 wwweee
5
2
1500
o0a2
45 aree
03
3
me
5
3
5000
o02
77 uzee
0
s re
5
2
Positive Control S0-1aminoacrid6i6n3e 75 ug1per plate
002
3r6y2 11
545
175
BackgroTuenNodrnLaalw;n C2o4dSelightly reduced; 3sModerately reduced
N4a=tEixotnr.eImnetleyrfreerduicnegd;preSceiApbistenatt;e; 6I=pOsbIsnctuerrefderbiyngprperceicpiiptiattaete
BioReliance
---
Study No. AAS52XL.502001.BTL
38
`Company Sanitized. Does not contain TSCA CBI
'H-25171: Bacterial ReverseMutationTest with `anIndependent RepeatAssay
DuPont-8868
Bacterial Mutation Test
Table 22
TSetsutdySNuubmsbteanrce Id: HA-A2S5Z1X7L1.502001.BTL
strain
+ masa?
ECxopielrsimSeenetdeNdo :: B02.5 X 10
VLeihviecrleMicrosomes +: Rtaettralhiyvedrro$f9uran Date Plated : 15 Jan 2002
Plating Aliquot : 25 ub CNgoncpeenrtrpaltaitoen NPulmabteer pRoerverptlaanttes
BackCgordeound
RAevveerratganets
_ SDteavnidaatridon
Vehicle 0o21 0
56 11 81
2
15
o012
s3 11
0
51
2
50
002
`5 11
0
12 1
7
.
150
002
38 11
03
61
5
2
500
o012
53 wwee
o
2 we
5
2
1500
0012
3s 1irpe
03
6 re
5
2
5000
0012
35 x1pp
03
7 ze
s
2
Positive Control 2-oraminoanthra56cene 1.01ug per plate
0032
Ek74 11
0
1
BackgroSueNnodrmLaalw;n C2oedsel.ightly inEwtremoly reduced;
Sre=dAubcseedn;t;
3eM=oOdbescruarteedlybyrepdruecceidpitate
Neation-Interfering precipitate; IPsInterfering precipitate
- BioReliance
-
Study No. AAS2XL.50200B1T.L
39
Company Sanitizad. Doesnot contTSaCAiCnBI
H-25171: Bacterial Reverse Mutation Testwith an Independent Repeat Assay
DuPont. 8868
Bacterial Mutation Test
Table 23
TSetsutdySNuubmsbtearnce Id:: AHA-S2Z51X7L1.502001.BTL Experiment No : B2
stLirvaeirnMicrosomes :: wNeozneuvia
DCealtlesPSleaetedded :: 125.1JaXn102002
VPelhaitcilneg Aliquot :: 2t5eturbahydrofuran
--
Cuognpceerntrpaltaitoen NPulmabteer pReerveprtlaanttes BacCkogdreound RAevveerratgaents DSetvainadtairdon
Vehicle o0r2
11 11
03
10 1
un
1
15
002
107 11
03
51
5
2
s0
0021
11s2 11
0
71
150
0012
.5 11
0
1u 1
u
.
8
.
500
0012
Fi7 mmee
03
7 we
10
6
1500
o012 c 12 1p
03
3 1p
13
1
5000
o012
51 z1ep
03
1
re
1
1
Positive Control meo1thyl metha1n40esulfonat1e 1000 ug per plate
0032 13581 a1
120
36
BackgroNuonrdmaLalw;n C2o-d5elightly reduced; 3=Moderately reduced
N4PeeENxotnr-eImnetleyrfreerdiucnegd;preScwiApbistenatt;e; GI=PO=bIsnctuerredferbiyngprperceicpiiptiattaete
c=Contaninated
BioReliance
~
Study No. AAS2XL.502001.BTL
40
`Company Sanitized. Does not contTaSiCnA CBI
'H-25171: Bacterial Reverse Mutation Test with an Independent Repeat Assay
DuPont-8868
Bacterial Mutation Test
.
Table 2
STetsutdySNuubmsbtearnce 1d:: AHA-S225X17L1.502001.BTL Experiment No : 52
stLirvaeirnMicrosomes : WRea2t luivver 59
DCealtlesPSleaetedded :: 125.1JaXn102002
VPelhaitcilneg Aliquot :: 2t5ecuxiahydrofuran
C4gonpceerntprlaattieon NPulmabteer pReevrerptlaanttes BacCkogdroeund RAevveerratganets DSetvainadtairodn
Vehicle o02r
1262 11
5)
10 1
16
5
15
o012
210 11
03
15 1
16
.
0
0012
11B2 11
-
03
15 1
1
2
150
0021
1n6 11
0
10 1
1
`
500
002
18 wwee
03
16 we
12
1500
002
112 1ipe
0
13 ie
n
1
5000
002
17 11pp
0
12 ae
1u
3
Positive Control 20-1aminoanthr81a6cens 10 u1g per plate
0023 152388 11
961
157
Backgro1u-nNodrmLaalw;n C2o-dSelightly reduced; d=Moderately reduced
N4P=sBNxotnr-eImnetleyrfreedruicnegd;preSc=iApbistenatt;e; I=POsbIsnctuerrefderbiyngprperceicpiiptiattaete
BioReliance
_
Study No. AAS2XL.502B0T0L1
a
Company Sanitized. Does not contain TSCA CBI
H-25171:Bacterial Reverse MutationTestwith an IndependentRepeatAssay.
DuPont-8868
Bac`tSeurmimaarly MouftaRteisounltTsest Table 25
TSetsutdysNuubmsbtearnce Td :i HA-M2S5I1N7L1.502001.BTL Experiment No : Bl Liver MicrosomAevse:ragNeoRneevertants Per Plate : Standard Deviation
Dose (ug/plate) 8 ma100 TasYS
V1e5nicie
T1e5s+ 63 ai0m:: 228 115ss: 1&
15050
e 1s331162:: 2265 1139:: 34
15S0000
B1r3oad I 2 1:520304 112s: 54
P5o0s0i0tive
211s4 251074:51222 116:: 404
Liver Microsomes: Rat liver 9
AISI 55:: 42 57:: 33 45:s 11 5074:2333
we2 wr 1154:: 22 1133:: 33 113:: 43 310:: 42
Dose (ug/plate) TASS Ta100 mass TIS) we2 vr
~
V5enicle
2237:+ 27 1a8m0s: 2278 2200:: 35 180:: 35 1156:: 44
15050
2270:6 65 s 19ss1104 1155:: 25 89:: 22 117:: 43
1550000
2333:: 37 a19m32 s1169 1162:: 33 76:: 32 115:: 22
P5o0s0i0tive
4230:: 8342Bi0s02:823 11699:+ 235 s7e:: 102 76163::1s4
VPeohsiictlieve==VePhoiscilteiveCoCnotnrtorlol
Plating aliquot: 25 ui
_
StBiuodRyelNioa.ncoAAS2XL.5020B0TL1
a2
CompanySanitized. Doesnot conTtSaCAiCBnI
'H-25171:BacterialReverse MutationTestwith an Independent RepeatAssay
DuPont8868
Bac`tSeurmimaarly MouftaRteisounltTsest
Table 26
TSteudsyt SNuubmsbtearnce Id : AHA-S2Z5X17L1.502001.BTL
iment No : 82
Liver MicrosomAevse:ragNeonReevertants Per Plate + Standard Deviation
Dose (ng/plate) TAS TAI00 TAISIS TAISI7 Wea uvrh
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--
Study No. AAS2XL.502001.BTL
43
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APPENDIX A Historical Control Data
BioReliance
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Study No. AAS2XL.502001 BTL
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--
BSitouRdeylNioa.nceAAS2XL.502001.BTL
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H-25171: Bacterial Reverse Mutation Test with an Independent Repeat Assay
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APPENDIXB Study Protocol
BioRelianee
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Study No. AAS2XL.5020B0T1L
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----------------
StudyNo. AAS2XL.50200B1T.L
54
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10 REFERENCES
Aames,tBaNg,sMwoiCtahtn,eJS.ealnmdonYelaa r,El.i(9s75).rMoetmheodTsafogrdctingecta.cMniosgieoon
Reh 3134736.
EhGreerni,cMhILco,lsMuntdaMtiaoinlRe,seWarJc.h 3(8139.7362).Motagntingwingty' reversionia
RRenegtpusutatinooirnosyn)GoefCnoPhinamiecceTseiocsnaslHfsoafmoPchoPiassrimaoensU(steCo.H.)CoSuf2b4Tdaoocce0cn8acStprReccooimesgnaepieendcsoofocfx
a Sri T8a20m 2i, po4ep ioRfti h24e,1596I. Co H`on c ow nJye13,p 1995, FeRdeegier
RIeogepmaariitoonnaofCPonhfraocse eonsHfaorrHrisnonUse(,CGHn)oosfiTcecyh:oiAcSl aRerqudirBeamerntysffoxr
--
NGSoevpneom4tboeirfcy2th1Te,eI1s9Ci9nH7gporfocPeasmsacoansiuayls1.6S,21899d7o.cFumoednteneRcoeagiseerndSdEfoArCEo1p6iIn5,
M Saci1.C1m20Aoamesne,,tB:eNs.y(i197olf6)3D0a0etcchoieonmroo:fdvcsarccainasogo.essrmo5.NesealgAecsadi.Stte.
fries
mMaogane,sJsChoS,E,dYammao,E.rso Aeeracl,BeXyr.of(13m9705).iDheteeocsti.osnPofrouo.cNnenagAecssa
SUATsIssi
MTareoV,iDMa.nsRdAensh,B1N31.7(51291135) ReidMethodsfor heSlmonell Mites
`OAdEdCeDndGuunidoetlihneeO4E71CD(GGeneutic iToxficool0ogy:TeBnag ocfCRheemvienM,optoiolneTdey),OENCioDt,h
PaisFem 1998
SWpeiellrmceaoen,enP. Npabiidmoa,sAn,TWAeLdEd,iDhJ.EcsahdGacolclWDP?Gh.t(eo9e90in)..CosmMpaaregieson,nof
Prac POTSO2001 190201 ge100( 11
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Prot SIOTS001 190201 Pagelof 11
@ BIORELIANCE
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Study No. AAS2XL.50200B1T.L
57
Company Sanitized. Does not contain TSCA CBI
H-25171: Bacterial ReverseMutation Test with an Independent Repeat Assay
DuPont-8868
APPENDIX C Information for Japanese Regulatory Agencies.
BioReliance
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Study No. AAS2XL.502001.BTL
8
Company Sanitzed. Does not contain TSCA CB
HanL2I5n1d7e1p:enBdaecntterRiealpeRaetveArssseayMutation Test with
DuPont8868
ReportofResultsofReverse-Mutation Testin Bacteria
1 Tester Strains
1) Procurement
Obtained from
| Date obiained
10 November 1998
tDhaetstirnaisnpelcotteidn storage
TALS3S
Dr. Bruce Ames | 11 August 1998
[aTTAAILSr S38 n| CoUlniovemrsiBteyrokfley
-- WP2 rh
RMON
MoafrIinndeusBtarcatlearnida
We gimton) | Aberdeen: Scotland
Storage
Froczing method
1luly 1987 1g peyrary 1993
--Theggeenneetti|c
condfaiyrmoetdwoen the
N Composition
Bacteria suspension
10mL
DMSO
009 mL
om ee Jee[] i] 21.)5S9ouMricex,StorageTemperature, te. of$9 .
26 November 2001 (Bich R656)
Name and model of| So-Low, Model
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BSitouRdeylNioa.ncAeAS2XL S2001BTL 59
Company Saniized. Doss not contain TSCA CBI
H-25171: Bacterial Reverse Mutation Test with an Independent Repeat Assay
DuPont-8868
(2) Preparation of $9
--| Rattus norvegicus, Sprague
Inducing substance
Administration
[w [Em e Le] Sm |Ame
[weight| 1871sB0at2chR76516)| (ggweighy
i
soo so
bodyweight
3. Preparationof Test Substance Solution
- CRer Te [on [me Tetrahydrofuran
`Aldrich Chemical
`Grade and/or
Gold Label,
`Stability of test substance in
Method of suspension when
test substanceisdifficult to dissolve
Sonication and warmingto 50C in screw-cap tubes.
Ee
Pe.
Study No. AA52XL.50200B1T.L
60
Beene 'H-25171: Bacterial Reverse Mutation Test with
sss
I ren |Srvow 4. Conditions ofPre-culture vm |_; |
E Soe n0 ms0008 eapn e
Frs---- shaking culture
--
deendofcultureto usefortest
Model and manufacturer of
<<
a8
New Brunswick Scientific, model G-24
Methodofshaking
(shaking type, speed, etc.)
Rotary (125 rev/min).
Tr-- Culture vessel (shape,
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200mL
--_
capacity)
shape: cylinder,
5. Agar Plate Medium
ro
(2) Minimum Glucose Agar
tee row
Mw E TowE o T ] [ove | woonorso | [Me ant mp]
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BSitouRdeylNioa.nceAAS2XL.502001.BTL
61
H-25171: Bacterial Reverse Mutation Test with an Independent Repeat Assay
DuPont-8868
6. Test Results- Judgementofthe results
[otgmen vege
Reason forjudgement and referential matters: Nopositiveresponse wasobservedwithanyofthetesterstrainsinthe presenceand. absence of Aroclor-induced rat liver S9. Referential matters "The vehicle and positive control values indicate that all tester strains were functioning correctlyandwerecapableofdetecting amutagen.
BioReliance
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Study No. AAS2XL.502001.BTL
62
Company Sanitized. Does not contain TSCA CBI