Document wgKp5Xq6wXqKxkDxR8kEL7xw4
MUTAGENICITY EVALUATION OF
1:2015 coc sAMeEsS SSAALUMOOIRnREETLLiTXeA/uMiIcChROoSsOoMnEe
FINAL REPORT
SUBMITTED TO: aSnh cCENoTEmR SAINT PAUL, HINNESOTA 55101
[3 BIONETICS
suBmITIED BY: KENLSIIT5N5TG1OT6NONBN,IIOCNHHEOATLRISTCOLSNA,NDLAINNCE2.0795
L8I PROJECT Mo. 20838 FEBRUARY 1978
00165
I. SPONSOR: 3M Company
1m. AMATERItAsLe*itication: T2015 Cov bouts fecetves: December 20, 197 C. Phystent Description: White powder
nLTM.. TrYrPoETocOoFLASrSoA:Y: owr-ATmoens Salmonella/Microsome Assay v. resurs
The results of his assay are presented fn Table 1. VI. INTERPRETATION OF RESULTS AO CONCLUSIONS
Tse! The test compound es ely yi examined: forJulagentc etivity Sebi, in a series Teed rats. pirnedsiecnacteoroforglainviesrmsm,icroTshoemaclomepnozuynmdewparseptaersatteidonds refcrtolmyAarnodclorin- the hTheerecowmapdunedityhaesr qteusatnetditaotvievreoofsearuiessiiioafticvoen.csewnitdreantcieonsofspuceh that aTcnh1ee" mitTocowaIlcdloyse-eir"nedicuntc.aed1 TphCheayssedisooslevoagsiF.caarbloeeleoefwmfpealcotcysoendcaetnftotrrhaetSihheoingehvtsdhjoautssetdieloesnvoemoli,fraItoesd oTchofemaporuRensedutltwasabsoilcffrtaohmcei0v.ta1etsituosgnctSooynsd5tu0ec0mteudvgerapenerothpTelacrteoe.msptoiuvned. fn the absence oTFheareastultisveorf atchetivtaetstisoncsoyndsutcetmedverosn 4th1e rceosmsaruined. In the presence
"Information was supplied by the sponsor.
by the sponsor, N.I. was entered,
If information was not {indicated
Gan[HB BIONETICS _--
-------- 00166
Vi. IERRETATION OF RESULTS MD CONCLUSIONS (Continued TrhoeaEtnyeltorecoRnmEpousSnhd,aiTt2ch0eo1s5encoaeCknhc1dnorhmoitidnSe:maornasatvriaotne gonednewtikc aScotnivirtyed
[HeBoI nenIcs
Stmttad by:
Study Director
2A opus SSSeOercREeat.irJoEinagzeaeantonerartchS,nePehi.tDs.
220%
Date
Revie by:
Sw rrviaircieernT ssicohf canI;es # 7
.
00167
232 B@3 oo
proTOCOL
1. pumsos TaTihcaetnivpmiuetrtypaobsIoenlioacf matihccitrsiovbasittaiulodnyaswpsaraseypatrwoaitteihvoanlasun.sdtewitthheouttestthemaatdeeriiaelionfoorfgmenaemtmiac
2 wes
A. Indicator Microorganisas
AingdePsrcorciepdtuiroen oonf psatgreai1n0.verification fs given in Standard Operat-
Salmonella typhimurion TThh1-8153375
TTTAah-s1is00s
Saccharomyces cerevisiae 04
8 Activation Systen
.
1. Reaction mixture
Component GSTlPouHdciou(smseo-pdthiopusmhpohssaaptlheta)t(edibasic) HH&eoenlosgenate 59 fraction 2. 59 homogenate
Fina) Concentration/al 1040$ fypmmooorll 03.38 1mmao5tl wm
Aa0du9lK.t0i0m0alaexccrogartdsiTunipgveertrnoatiatnnhdteucepwdraoscbeydpurArerep.oacrloeofdr A1nf2er5so4naftiSvpaer1a.dgauyes-(D1p5a7rw8il)oe,ry $gPBrr9yoadmmsuseactmtphp.rolodetsseidwneesrpceerricaboeidde1d1i1n1b1yt(a8l1roxaTnemdcuhsnrbieeclraaltainvDdeataaPsesAoaEny/ePdfAaStEfohracetAeiLrv1i1tI5y.
ion [3BIONETICS
-
00169
2. MATERIALS (Continued)
C. Positive Control Chemicals aThcetivcahteiaoincalasssauyssed arfoer gpiovseintivine TcoanbtlreolTs oifn tSheectnioonnacVt.ivatRieosnulaesnd
0. solvent
pErietphaerre dsetioocnkizseodlutwiaotnesr oofr sodliimdetmhaytlesruilaflosx.ideAl)(DMdSiOl)utiwoanss uosfedtestso SVmeaentcte.triioeanslpVsl.owyeeRrdeesaumnladtdse.fsin ceointcheenrtrdaetiioonnizeadrewarteecrordoerd DMiSnO.TabTlhee 1soolf-
3. EXPERIMENTAL DESIGN
A. Plate Test (Agar Incorporation)*
BFtAoooplrrptreonsnxotinrmaaaacigttnaeirlvwyaetsriu1eop0n%plaedtcmdeeeeslndtltsse,tdofartwosimetplhaearanasbttioeov.4teirtnndeiosgstahentdtlucebauvelesttlursracecooenfotfaotifheneaichnhtigestsi2itn.dd0icinocenaa.:1-
Ppoouurnedd tests,
voaevtreerle`atashdteded4surdtfooasceetshleeovfeclosnsteeloneftcsttihveoef
taetgshater
Cpahlpepamrtioecp.arliawtIenereatcutabidevdsaetdiaotnnod
hathloeimqougaoeptnpartooefp)rriewaaatcsetioatndudbmeeidsxttuworieteha(c0h.c5elolmfs1.
Just prior tchoentaaicntiinvgattihoen
to" pouring. an 9o,v0e0r0laxy g"tluibvees,r
W3himcihniwnearle. atghaern mpilaxteed', aanndd athlelowceodntetnotssop1oiuariefdy.overThteheplsaetrefsaceseroef
incubated for 48 hr at growing on each plate.
37C D4
and scored for the yeast plates were
number of incubated
colonies at 30C
bdos(oficnlrooirenSeccaedtcc.ltatyicivtoainT`tvhaiaceVott.inicv)ooennRcepvaseoenunsrdltietrtsa.ir3tvu7ienoCnPwscoihst(eoihamftciietcvaaiaelcvllhastainacaosdhneneds)nsyio.ctlfahvoloerssneta3crt-eoh5nagttidrvaareytensquuiisnarinenTdgambelbtteehete:hn|
BmpCeurettorhadtogudaec.inenicdCSleofatnmseescetssadubislapoleefknyscrliheoesnnmpiioatcnsrassoleasssyasmk,uinsnoiebwnsungtattnnohdoetbceeisrntmtauatntihdneaargdpselunabAtsnaetenisdtasusatcegeasadr.rcihnyIoCanghrceaaonrsristsneoeerdslaostianoovonfet these Classes should be screened In a suspension aseey.
3BIONETC':S
. 00170
3. EXPERIMENTAL DESIGN (Continued)
8. Recording and Presenting Data
Tpahnredintnreuedspboefrrotsremdso.foncToahloepnsrieiensrtaowountd.aetaachTwheeprlearteeasunwlatelsryezeadrceouipnnrteeasdecnaotnmedpdutreaerscoprrrdeoevgderraotmn
eamnptlsoy(eodr vided as
icrnoenftvehereretnaacsnestasyp.ofionrtTsh.De4)poOpsteihtreirvpelraaetnledevfasonortlveedanactthacioannrdteircopalrtsoovrairdesetdprraoio-nn
the computer printout.
4. EVALUATION CRITERIA
aPsluasstpeeetndtoeefdstsiednlaetcaaticsvoeenmsiaisgsoatlridofploadvtieerrselcatsye.redeevdeBrewtciaatnushtepcootplhuoelnaytteicsootnusncthsoefmoimbcutataliannaetnddceflrtlhosem conaectlculursre.adruerAiilnntgchuobtuahgtehedithniecnsuebtahtfeieoaontvuerrpeelsraiyoodff,ortht2ehedaasytssea,syt arniedsduacseefmeitwqhuecaenlqtluiatndatitivitivasetiiooninsn otfatirveesulstuss,pentshieoyn ptreosvti:de certain advantages not contained in a quanti-
+ TtnhooenraescmtpalliolncatnriuenapgbleircDNaAto.ifngcelDlNA,diwvhiiscihonsispoefrtmietnsmpoorteensteinaslitimvuetagtehnasn To2hveedarylcsao.ymbipneerdmitisnccuobnasttiaonntofexptohseurceomopfountdheanidndtihceatocrellcsellisn tfhoer A. Surviving Populations
ntPiulomanbtseerotfoefstthceepltrleossctesdcuuhrrevemsiivciadnlog, cnthohetemicpsaeulrrmviittvrienaegtxmapeconttp.ulqautaAintotniltooawntictohnoencotefrnettarhtae mcioesnnttruoslpulaalptlleyaste.riesduAcetesdsheinbgtyhiaclsolonymceetnhtferraatcsitaoimnoesn,. asthOetuhrsautrpvrioonvtionctgohlepopnnueolgraamttaiilovlney edhmeiptglehoreynssitneosdfevbetyrhaelssuebdjodesocestseisvreabnecgiriningtgersioeavl.eercte2dorto3 slhoogw cso1nicgennttrtaotxiiocnist,y tahse 8. Dose-Response Phenomena mTiihmgephotrdteammnootndsictfrryaittiedoroniseoo-nfreidsnopsoene-ssretealbraletisesudhlitnisgncrfmeouartsaegsaeniimncuittmayug.teanntAwcofouaulcndttosbreisthtahaetn
[H siIoONnEeTInCc:s
.
0171
4. EVALUATION CRITERIA (Continued)
B. Dose-Response Phenomena
isceilteyctiaorne roefladtoesde)s. concentration, no
thIatf tahree increases
htoioghelsotw may be
d(ousseualislyfamrutalgoewneircitthyanaand observed over the ose
totxoxi-c range
tsoexliecc,tedt.he Ctoensvterscehleym,icailf tmhaey lkoiwlelst adnoysemuetmapnltosyetdhaits ahrieghliynduccyetdo,-
and the compound will not appear to be mutagenic.
C. Control Tests
:
aePsxospsaeiyrtsiivmeeanntdanadmndutnacegogenanstsiisvttehoaftcodnriterrqeouclitr-eaacstmsieantygsabmoulatriaecgebcniosontdfruoacrntsenfdoonrawmciatttihivoanteiaocinnh
ascotlivveanttioinn ponents.
atshseayosv.erlaNyegaatgairvetocgoentthreolrswictohnstihset oofthetrheestseesntticaolmpcouonnd The negative control plate for each strain gives a
croenfterroelnceaspsoaiyntisto cwohnidcuhcttehde ttoestdedmaotnastraarteecotmhpaatretdh.e are functional with known mutagen.
tTehset possyistteimvse
D. Evaluation Criteria for Ames Assay
.
ptBoeescsiattuiscveheemtiehcfeaflepcrtaosrceeadsrueermeiisqnuhaeunrsteeindttalttyoivseeu,vbajletuchetaitvecerittahneedrimaaurteuasgbeeadnsietcdoitpdyreitomefarrmiitlnhyee
fonollaohwiisntgorcirciatleridaa:ta base. Most data sets are evaluated using the
1. Strains TA-1535, TA1537, and TA-1538
cIhfemtihceal sotlhvaetntprcoodnutcreosl concentrations with the
avaplouseitfisvewidtohsien lowest increase
rtheesponnosremaloverrangteh,reea equal to twice the
solvent control value is considered to be mutagenic.
2. Strains TA-98, TA-100, and D4
cIhfemtihcealsotlhvaetntprcoodnutcreosl concentrations with the
avaplouseitiisvewidtohsien highest increase
rtheesponnosremaloverrangteh,reea equal to twice the
mcsuootnlatvgreeonnlticcv.oanltureFoolrforvtahlseutsereaifnsostrraTTiAAn--s91,800 tahanendd Dd4o2s-e3i-srtecismopenossnisdteehreeidnscotrloevaesbneet
should start at approximately' the solvent control value.
[HoonETICS
2 00172
4. EVALUATION CRITERIA (Continued)
0. Evaluation Criteria for Ames Assay
3. rattern
dpBaeerrceianvutesadel
fTsrAto-rn1aSi3nt5h(e6a-ns4da6m)eTAa-pn1ad0r0ebnetcaaarluesebstoTrtAah-i1nd5e3r8(iDva0en5dd2)Tf,Aro-nt3hetarhreee
bsoatmhe is a
ptbwauotitlestrtinrnaiinsrsedsouofnudgahatn.csyetiInrtetsihpseondamlisctoroobatinhatelicsiaapsmasetaeym.duttahIganetngieafnnaedraslgucihvtehnae
ctseotsnrtvasei,"nr,sefet.go.fW,ilTtlAh-is1g5e3n7re,erlaarltelisyopnosdnhodispstoofisainmnuoattcatgieevnxaptieincotnneodnt)ae.csttisvWaht(iiTlohene
tshiemyilacranrebsepounsseed decision.
ptaotteenrhnasncaerethenotrelrieaqbuiilrietdyfoofr
aalnl
emvuatlaugaetniso,n
4. Reproducibility
ciIafnmintaioatlchbpeeomsiicrtaeilpvreopdrtuoecdseutdcedsiantaaoTnoreseesoprsoingsBneoirfefincaanadcdei.stiinognlael treusnts, tthhaet
tfThhaeecsteoprrscerciemtdaeiyrnigaenctarerireteairpniptaloieadaretfoinnoattlheaebmsvaoajllouurtaiett,iyonaonfddescioittsuhiaeotrni.oenxsteHanonwudeavteiarnr:ge
acpetrdieuosrneen.ctaendAbsteotmhoaerieddfatithraomslbeyaseiesntdaiibsvliidisunhacelrdse.asneodt, ftahmeilcirairtewriitah fothrisevparlouc
E. Relationship between Mutagenicity and Carcinogenicity
rTIeetcsotmgunisistzednb,oet heoamwpedhveaefrsi,inziettdhiavtethcatotersrtethleaftoiArvmceehseanmSdiaclfmauolnncetclialoranc/aiMlniocgrreeonlssao,tmieonIsPthliaptises
sohfahvoewcoabmnepeanreaxtt`irdveeemmoenltsyetsragttsoeoddonbceot3rw0re0eelnacthtiehomenisceabletstwwoebeyenndMrpcoeCisanuntlnst.setoTfmahi).ecrroe(bs1ui97la5tl)s
mutagenesis tests and in vive rodent carcinogenesis essays.
rAelplorte.vaalrueatiboanssed activity.
aonndlyinotnerptrheetadteimoonnstorfattihoen,daotra
lparceks,enotfedmutinagetnhiics
[HBaIONETICS
.
00173
ReFereNces
cGAmaeernsci,icnroeBgyNent.sestaH.ncdCaMmmuut,taatJgi.oennanRwedist.Yhas3t1s,hseakHiS,iTE.lo(H19n75e).liMeathsoomdsesnaefomomrseodntest:escutriendg
ctaarcmin,oge3n.s aC5rtm,itaEginsta1n mtheaS,aisasnndeolleas/,acr8is.one17t5es)t.:
Chemicals. roc. Mat. Acad. Sere T9ETIecsine.
DAestsyecotfioofn
[HesoioNETICS
.
00174
5. STANDARD OPERATING PROCEDURE
AT data will be entered in ink (no pencil).
tAlhlroucghhangtehse cohrancgoer,recatnidonasn ienxpelnatnraiteisonwfilolr tbhee mcahdaengweitmhusta sbeinwgrliettleinn.e
Aslhlowncaolncudlaattaionrseco(rwdesi.ghts, dilutions, dose calculations, etc.) will be
AT) data entries will be dated and initialed.
Amlalnuallsa.boraTthoersye moapneuraaltsiownisllwiblel prbeesewnrtititneneacohutlabionrasttoarnydaarrdea.protocol
Dfeievdi.ations from any established protocol will be described and justi-
Data data
will books
bweilsltobreedreivniewbeodundbyftohrem
a(pnportoepboroikasteorSecbtinidoenrsH)e.ads.These
bound
Cofheemnitcaelrsingsupbemristotne.d for testing will have date of receipt and initials
uLsoetd niunmbaesrssaysfowrilalllbe rreefceorrednecde. mutagens,
solvent,
or
other
materials -
tAnaiimnaeld ment.
osrudcehrst,harteceeaicphts,aniamnadl
identification can be traced
All animals on study will be properly
will be to the
recorded supplier
identified.
and and
mainship-
Abecoppeyrmoafnenthtelyfisntaolrerdepionrtthpelusarcahllivarlawsdyasttaemaonfd LsiutptpoonrtBdioonceutmiecnst,s wIinlc.l pCeurrsroennntelcuirnrviocluvlead viintateheansdtudjyo.b descriptions will be maintained on all
aSnadlmponKella 101strpahiennsotywpielsl. will be used in chemical
be Onrloyutianpeplryoprcihaetcekleydscfroreetnheed evaluations.
shitso,ckuvcru,lturrfae,s
[Haonercs
Utton
0
cCo17s