Document 5kJx9GgrBVwadgqeJog3Z9wk4
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nl
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SRPT T-766(
AR 826 - 1200
Wk MIN 314022208
g
ER33o2n2
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PERFLUOROOCTANSSULFONYL FLUORIDE GO Tlie
BACTERIAL REVERSE MUTATION TEST
=
Shomer 33MMCCoepnrtrsteToxicology BSutirladitng2202.03 UNsNa351333220
CONTAIN NO o m
BosarchLaboratory HuWnatlilnegydRoonaLdif SciencesLt, AHCulanominnbbgaririoyndge FEESNGRL4A5N5D
Reporte2d3October 2002
Page 10629
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MIN 3141022208
coNTENTS
Page
COMPWILTHGIOODALANBORCATOERY PRACTICE STANDARDS...
3
TEST SUBSTANCE wr DEVIATIONS FROM PROTOCOL.
9 es 18
TABLES 1. Resultsoad with S.phanTASB: ct | (aging)
ic 18
2. ResultsobtainedwithS. typhimurium TAS: test2,Withpre-incubation ..........cuwee. 19
543... RRReeesssuuullltttsssobobbittaaniineneedddwwWiithth.hSp..ppthyhiphimiimmrurtiuumn TTTAAAI1IS0O30DS::teseststt2|,1(wr(iaatnnhggepe-rS-eiSinnnddciinungbg)a)tion vocvev rcvrrvrwrorormrorne: 222120 67.. RReessuulltssaobbttaaiinneedd wiitthh5.. ppphhiimmriunTTAAISLSST:: tesst21,(wailnhgperne cdubiatinongv).r... 2234
581..0. RRReeesssuuullltsss obobbittaaniienneedddwwwiiitthhtSh.. cocooyllplihWiWmEu2rPiioumrAAT/A1pK5KM3M7II:0Ot!Les((tC2CM,IwM9iB1t)hO:p1treees)s-ttin2|,c(uwbaiatgthieop-nrie.n-.nd.c.iu.n.tg.a.i)o.n... 222567
APPENDICES
2. EyeResearchCentre GLP Compliance Satement 2001 vr
29
00003
5
sfoniiign COMPLIANCEWITHGOODLAORATORYFRACHCE STANDARDS e Be Ri nite ningt Stnosm.
tinpte HH gs Schrrn Ae ed SHESneer atISAT,
Psaipnses we pyar In line with normal practice in this type of short-term study, the protocol did not require analysis of
K"em1entwrindeLcat z ey
i23..0ckber. 200 }
pan
000004
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MIN 314022208 .
QUALITY ASSURANCE STATEMENT
The following inspections and audits havebeencarriedoutinrelationtothis study:
Study Phase Protocol Audit
PSFOroorPcmreuosppatBroasnsioeandndInTsrpeeuctmieonnts Pie Scaring Report Audit
Date ofTaspection 16 October2001
3115 JOacntuoabreyr22000021 24Cctober 2001 4February 2002
DatofReporting 16 sober2001
1351 JOcataorbeyr22000021 24 October 2001 $ February 2002
PDirroetcotcoorliAnuddCito:mpAannyauMatnoafgtehemeprno5ttodcoilsfoirdthasbosvteu.dy was conducted and reported to the Study nPrdocreespsetBitaisveedprIonscpeedcutrieonse:mpAloyoerdsboonuthstheytpimeofthsisndstyudwyerweascairnepdroogurt.esTihnesspeectwieonrs coofnrdouucttiende 2ndreportedtoappropriateCompanyManagemensts indicated above. cRepoorntdAuudaintcd:rtTephoeirtdreedpootrht hasStbuedeynDiaruedcittoerd abnydCtohmQpuaalniytMyaAnsasgureamnecnetDespairndmiecntte.d aTbhoivse.audit was "rTehseumseotfhotdiss,sprtoucdetdyourreesflaencdtohbsreerdviattiao.ns were found to be accurtely desibed ad th reported
AGnrgoeulpaManMasglearn,ings, B.Sc, Me. PhD. MRQA,
Date
Deparment ofQuality Assurance,
Huntingdon Life Sciences Ltd.
poatas
000005
4
RESPONSIBLE PERSONNEL
KSetnundyetDhirMeactyo,rBSe.,C Biol, MLBiol EScliieznatbiestth Farmall, B.Sc.
MIN 314022208
pani
yrs
. 5: . 000006
MIN 3141022208 SUMMARY In this in vio assessment of the mutagenic potenisl of Perfuorooctanesufony! flucide (POS), ahinsdtidTiAnLe0dDe,penadnedntaautxroytprtoopphhiacn mduetpaenntdeonftSamlumtoannetlloafbEpshcihemrziicuhmia, scioali,nsstTaAiInS3WSE,2TuArIAS/3Tp,KMTIAOSLS (CM891), wereexposed tothe testsubstance. "ArTowcolionrd1e2p5e4nd-einntdumcuetdartaitosn(t5e9stmsiwxe).reBpoethrftoersmtseidi ntphvresaopernlecinevcaunbeadtaidobnsstaeogefninlciavieenripgrhetpvaersasteilosn.s from `Cmountcaetniornattieossn.s oThfiPserisftlhueosrtoaocntdaanrcdsullifmointyclonlcoerntirdaeti(onPOrSeFc)omumpentdoeSd0i0n0thpeg/rpelgautleatwoerryegutiedsetleidneisntthhaet his assay follows. No siofgtonxicisty wereobservedtowardsthe estersain n eithermutation est. No evidenceofmutagenic activity was seen at any concentration of Perflvorooctanesufonyl fluoride
(POSFi)n either mutationtest.
"oTfhtehceonlcivuerrrpernetppaorsaittiionvse.controls demonstratedth sensitivityoftheassayand the metabolising activity sTthoiswecdonncoleuvdieddentchaeot,fumnudteargetnhiecatcetsitvictoyndiinttihoinssbaecmtperlioaylesdy,stPeemr.luoroocanssulfonyl fluoride (POS)
090007
Sos 5
MIN314022208
INTRODUCTION
ugThciirslidereespo:(rFtOdSsPs)riibneas bascttedriyaldseyssitgenme.d tTohaesssetsusdythweamsutcaognednuicctepdotientciaolmofplPeeflwoirhoohceanoeluolmoinoyg| ReOvBeCrDseGMuuitdaetliionnesTefsor,GtuheiTdeestlini4gn7o1e.f Checls. 1997) Genet Toxicology: Bacieral mEuCtaCtioomnmitsetsiionnbacDtierreicat.ivNeo2.0L001/336/25/7E.C Aunex 4D-B.13/14. Mutagenicity - Reverse UmuStaEtiPoAn e(s1t9.98E)PHAea7l1t2h-CEf9f8e.c2ts47Test Guidelines. OPPTS 870.5100 Bacterial reverse JGeanpearnalM,inAigsrtircuyltoufraAlgrPircoudltuucrtei,onFBourrsetauy.aNnodhSFaisnheNroie.s,42(010985) Notification of Director EiJosienit NDoi.vc1t2i7vaesndoKfJiEPkA,yNooM.kE2 Wa31aOncdtJobMerI1T9973)1, October 1997) Kapsn No. 287,
IMHW Genotoxicity Testing Guideline, PAB Notification No. 1604 (1 November 1999).
Official Notice ofJMOL. (8 February 1999).
TrehceommmeetnhdoadtidoensscroifbtehdewUansitaeldsoKidnegsidgonmedEntvoircoonmmpelnytawliMtuhtIagCeHn(S1o9c9i5et&y (G1a9t9e7h),ouasend1faoll1o9w9e0d).the MsTapherecoiainnllavynisderAlmtceeicsdhns1i9qi8un3e)sdcoessfbcSlraielbsmeodtnhbeeylmlAuatmpaegsehnnicdiefhmfiesctc1oo-fhwaoetrskteeksrtsssu(uAobmssaenstc,ae.Mn0ccNbCeaondmneltaeynrdm5iYnapemalbsyaikeixspo1ts9i7n5g, iocarpinagbnlacel ohfaiossfipytinhnhiaeessfiibunsndicelntpgieoenhnde,enWetshsfeeonrtnmitalkheeasmsiespnloiacaneisi,aripesroeipxonpro,tsieobdnao0fhteahemtupaognpeitantsi,orine.vnerTsveodhmauateaesiroenissne1ef1he0o maerdeiuvmr(tsanuantpdasle,ereamiditelhyndbiotteidn,tsinbcey hcheessteasialeisdtaoeraoswoandcafpaor cof boiosolnyaoehsin),sisdeefesnd AmisurtteaqnuteirsesadnfhbosraosgnfreodEiwostcnhqh.seirumiiclheairapcrionlcihplaetshsrainlcsapbabole odfseycnrtihbeesdibsiynGgreteenm(i1n98d4)a,idT,hips spystte,emwphliocyhs Tihnpteohstiahmicrnlli.uwmAelsdlectiahnmsrsythaaadindvsietaoendaedleifciimcvuietenaettliilonncnsotoutam,atwlhriDecNnhdAearlrtlophwaestmrremosocererespsieasrsmieIatnbviedliittyooomfnutteahsgesabeossf.atnecThehmse pSeonsssietsisvitayp1l0assommide m(uFtKaMgeIaOsL., which introduces ancorprone fopair process, esling in nreased
wid
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MIN 3141022208 :
Manysubstancesdo not exert a mutagenic effect uni they have been metabolised by enzyme systems anobtseanvcaeilaanbldepirnestehencbeacotefarisaucpeplll.emeThnetreedfolriev,ertfhaecbtaicotner(i5a9amnidxt)esptrseupbasrteadncferoamraitnscpurbeavtieoduisnlybtortheattheed with substance (Aroclor 1254) known to induce a high levelofenzyme activity. "oTnh3ep1rJoultyo20cw0oa1lsanadpbpryotvheedSbtyuHyDiurectnoroLtnif1e5SiOccitenonbceersg2M0a01nd.ageomennton 18July 2001,bytheSponsor The study was conducted at Huntingdon Life Sciences Ltd, Eye Research Centre, Eye, Suffolk, 1P23 7PX, England. EExxppeerriimmeennttaall sctoamtpdlaettei:on21daNtoe:ve31mbJearn2ua0r0y12002.
LA ELE]
000009
18:
Identity: Appearance: Storage conditions: Lotoumber:
Expiry date:
Purity:
Specific gravity: Date received:
TEST SUBSTANCE
MIN 3141022208
Perflucrooctaneulfony| fluoride (POSF)
Clea liquid
Roomtemperature
wo
b orai tion 95.5%
assumed stable for
cals
14 June 2001
)
COMB
000010
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EXPERIMENTALPROCEDURE
MIN 3141022208
BACTERIAL STRAINS
`The following strains were used:
S.DphimuriumTA1S3S: contains histidine missense mutation (hisG46) but is also deficient in a
DonNtAhereceplalirwaslylst(efma(m7utBat)ioann).d
has It
a is
defective reverted
lipopolysaccharide coat by many agents causing
base-pair substitutions, but i not sensitive to frameshift mutagens.
S Ophimurium TAIO0: is the same as TAIS3S but contains a resistance transfer factor conferring ampicillin resistance and increasing sensitivity to some `mutagens (plasmid pKM101). In addition to base-pair substitutions, itis also able to detect certain frameshift mutagens.
S phimrium TA1S37: bears a histidine frameshift mutation (K5C3076). Like TAIS3S, it is defective in 2 DNA repair system and lipopolysaccharide coat. It is dseenlseittiiovneotfoa asgienngtlse bcaasues-ipnagir.frameshift mutations involving insertion or
. typhimurium TASS:
acondteafiencstiavneotDheNrAhirsteipdaiinresfyrsatmeesmhiafntdmultiaptoipoonly(s4aicscDh3a0r5i2d)e. coAagtaibnutitahlasso `contains the pKM101 plasmid. It is revertedbyagents causing deletion osifmptlweoalakdyjlaacteinntgabgaesne-tpsaciarussi(dnogubbalsee-fpraaimresshubisfttitmutuitoantsi.ons), but not by
E coli WP2uwA/pKMIOI: (cMm391)
`contains an ochre mutation. It is reverted by many agents causing A-T base-pair substitutions at the #pE locus or by G-C base-pair aslusbostidteuftiicoinesntinintraansDfeNrARNreApaliorcisyeslsteewmhe(r7e Ai)n ,thae ncdhriosmomsoormee.reaIdtiliys reverted by certain mutagens then its parent strain WP2. It also contains the pRMI01 plasmid.
The strainsof 5. pphimuriuamwere obtained fromthe National CollectionofType Cultures, London, England.
The strainof. coli was obiained fromthe National CollectionsofIndustrial and Marine Bacteria, Aberdeen, Scotland.
Batchesofthestrains were obtained from masterstocks held inliquidnitrogen.Thetestbatcheswere taolitqhueotcsoulftunruetsriaetnt8b%rovthivcualtsureascarynodprweesreervsattoirveed.atE-a8c0hCb.atDicmheothfyflroszulepnhsotxriadien(wDaMsStOes)tewda,swahdedreed applicable, for cell membrane permeability (r/2 mutation), sensitivity to UV light and the pKMI01 plasmid, which confers resistance to ampicillin. The responsesofthe strains to a seriesofdisgnostic mutagens were also assessed.
For use in tests, an aliquotoffrozen culture was added to 25 mlofnutrientbroth and incubated, with shaking, at 37C for 10 hours. These cultures were intended to provide approximately 10" cells per ml, s`wuhrifcahcewoefrpelamteeassoufrendutbryiesnptreaagadrinagnadlicqouuonttsin(0g.1thme1r)eosufltaan1t0c+oldoinliuetsi.onofthe overnight cultures on the
Ginusg
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MIN 3141022208
POSITIVE CONTROLS In the absenceof$9 mix
Identity: SCupApSliNeor:. PLuortitryu:mber: SAoplpveeanrt:ance: Concentration: CIdAenStiNtyo:: Supplier: LPuorittnyu:mber: SoAlpvpeenatr:ance: Concentration: Identity: SCupApSliNeor: PLuortitnyu:mber: SAoplpveeanrta:nce: Concentration: Identity: SuCpApSliNeor:: Lot number: PAuprpietayr:ance: SCoonlcveennttr:ation: Inthepresenceof$9 mix CIdAenStiNtoy:.: SLuoptplniuemrbe.r: Purity: SAoplpveeanrta:nce: Concentration:
Sodium azide Si2g66m2a8C2h2em8ical m7i7nH.09T9.95% `DWMhiSteOp(oAwldderirch, A.C. spectophotometric grade) 05 pgpflorasttaiens TAIS3Sand TAL0D
99-0A4m5in9oacridine 1S0i6gFm-a0C6h6e8m1ical >om% DYeMlSloOw (pAolwddriecrh, A.C. spectrophotometric grade) 30 g/plate forstrainTAIS37
2Nirofluorene,
607-578 Aldrich Chemical Company
.
80501-24227
B9e8i%ge powder
DVMkSgOl(Allfdorraisctthr,aeAin.TCA.OSspectrophotometic grade)
32608-8-F5u3r7yy3<S-n2ifturryo) acrylamide. (AF-2) PWaAkEo1P1u5r1e ChemicalIndustriesLid. 9R6e-d1p0o2w%der D0.M05SgO (lAldprifcaoh,sAtt.raeCin.WsEp2ecut0rroAph/optKoMmeIt0r1icg(rCaMdEe9)1)
26-1A3n-i1n3oa8nthracene: `5A2l2dr3i-c0h2C4hemical Company G9r6e%en powder 2DMglSpOla(tAeflodrriscthr,aAi.nCT:SA.LSs3p5ecrophotometric grade) 10 g/platforstrainWP2u7AIpKMI01 (CME91)
Poh
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Identity: SCuAppSliNeor:: PLuortitnyu:mber: SoAlpvpeenatr:ance: Concentration:
Benzofelpyrene 50.328 `0A7l7dr8i-ch10Ch5emical Company 98% YDeMlSlOow(Aplodwrdiecrh,A.C. spectrophotometric grade) 5 ug/plateforstrains TA1S37, TA98 and TAI00
PREPARATION OF $9 FRACTION
Sspeexci:es: SStoruaricne:: Weight:
RMaatle SCphraargleuseRDiavwelrUeyKdeLrtid.ved <Swg
$9frawacsptrepiareodfrnom agroupofca 10animalsaccording tothe method describbeyd Ames, MArcoCcalonrn1a2n5d4,Yaadmmainsiasktier(e1d975a)s.asiMnigxleedintfruan-cpteirointooxnieadlasienjseycsttioenmisnicnotmheoiralt altiv2edrs owesreaosftgi5m0eu0latmed gby bceordvyicawleidgihstl.ocatOinontahenftihftehidliavyerasfaesreptiincjaelcltyiorne,mfoovleldo.wing overnight fasting, the rats were killed by
The following stepswerecarried out at 0-4C under aseptic conditions. The livers were placed in 0Fo1ll5oMwinKgCIpre(p5aramtlionK,CIthe: h1ogmogleinveart)ebweafsorceenbteriinfgugterdanastfe9rre0d 0gtof0oarP1o0ttmri-nEultvees.hjeTmhehosmuopgeemnaitsaenrt. fraction (59 fraction) was dispensed into aliquots and stored at -80C or below. Each batch of $9 fractionwas teste forsterility and efficacy.
Dateofpreparation: 17 July 2001 (est 1); 15January 2002 (est 2)
PREPARATION OF $9 MIX "The S9 mix contained: $9 fraction (10% viv), MgCl (8 mM), KCI (33 mM), sodium phosphate buffer pH 7.4 (100 mM), glucose-6-phosphate (5 mM), NADPH (4 mM) and NADH (4 mM). All the cofactors were fler-teilised before use.
FORMULATION OFTEST SUBSTANCE h"Tehxeanseo.lubiIltiwtyaosftihnesotluebtlesuibnstDaMncSeOw,asetahsasneoslseadnida adciemteotnhey,lasnudlpwhaoxsidken(oDwMnStOo),beetihnasnoollu,blaeceitnoneetearn.d a`tAtlhtihsocuognhcseonlturbalteiionnwheaxsafnoeuatnd5t0ombge/tmo,otthoexvictoowlaorfudhsetmhxeaefneestrseyqusitreemd.toIatdwmaisni,sthteerretfhoeret,esdtescuibdsteadnc0e administteher tet substance by direct addition without the useof asolve. All concentrations cited in this report are expressed in terms of the Perfluorooctanesulfonyl fluoride (POSE) sample as received.
Lrnnue
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MUTATION TEST PROCEDURE
First test(range-finding)
``TchoencteensttrsautbisontaonfcetewsatssuabdsdteadntceoctuelstteudrewsoasftSh0e0f0ivpeutge/spltaetrest(roabiinasiantesdebvyenacdodnictieonntroafti2o.n8s.iTlhoef thhieghteesstt
sausbssatyanfcoel)l.owsT.hisiThethoethsetrandcaorndcenltimriatticoonnscewnetrraeti2on50r0ecpogm/pmleantdeed(1i4n
the pu),
regulatory guidelines 1750 pg/plate (10
this ul),
`c1o2n5t0roulgslapnladttehe(0a.p7ppruo)p,r7i0at0epuogs/ipltaitveec(o0n4trpoll)s,w3e5r0epaglpsloiantcel(u0d.e2d.pl) and 175 pigplate (0.1 wl). Untreated
sFoollultoiwoni)ngtothaeiratidgdhittigolnasosfvtehseselasb,ov0e.5amlliquSo9tsomifxthoer 0t.e5stmslub0s.t1anMcep(hoorsp0h.1atmelbuofffepros(iptHive7.c4o)ntwraosl
amdidneudt,esfowliltohwesdhabkyin0g.1bemfloroefaad1d0ithioounor fbac2temrlioalfacuglaturrec.onTthaeinmiinxgthuirsetsidwienere(0i.n5cumbMat)edaantd3t7ryCptfooprh3a0n
(0.5 mM). containing
2T5hemlmixmtiunriemsalwerageart.horEouagchhlyPesthraikednisahndwaosverilnadiidviodnutaollpyrelvaiboeulsleldy
pwrietpharaeduPneitqruedicsohdees
co`crornecsenptornadtiionng.tPolataessheweetr,iedaelnstoipfryeinpgatrheedwciotntheonuttstohfeatdhdeidtiisohn.ofTbharceteerPieaitrndoirdsehretsowaesrseeussstehdefsoterreilaictyh
ohfoutrhse.teAsfttseurbtshtiasnpceer,Sio9dtmhiex aanppdesaordainucmeopfhtoshpehbaatcekbgurfofeurn.d Abalcltperliaatleslwzewrnewianscuebxsatmeidanted3a7nCdfreovrecrata7n2.t
colonies counted usinga Domino automatedcolonycounter.
A`cnouynttsooxribcyeftfheectasobfstehnceetoesftascuobsmtpalnecteewboauclkdgbreoudnetdebacctteedrbiayllasauwbs.taInnttiahlereadbuscetnicoenoifnarnevyetrotxainctcefofleocntys
ttohxeictoepfcfeocntcsewnterraetioobnsneorrvmeadllayulsoewdericnotncheenstercaotnidotnemsitgwhotubledbcheotsehne,saenmseuarisntghtahtautsesdiginnsothfebfaicstte.riaIlf
sinhhoiublidtiboen oabrteapinreeds.enItfaptretchieptiotaptceownceernetroabtsieonr.vedIdoenaltlhyeaplmaitensiamtutmhoefetnhdroefetnhoen-itnocxuibcatcioonncepnetrriaotdi,onast
least four Director.
non-precipitating
dose
levels
should
be
obtained,
unless
otherwise
justified
by
the
Study
Second test
T5h0e00segc/opnladtteewsatwsaagsaeinnechxoascetnraespteahtotfopthcoenfciernsttrtaetsito,ne.xceptthatonly fiveconcentrationswereused.
STABILITY AND FORMULATION ANALYSIS
`The stabilityofthe test substance and system were not determined as part
the stability and of this study.
AhnoamloygseisneoiftyaocfhitehveedtesctonscuebnsttraantcieoninWtahse
test not
`performed as partofthis study.
nal BE
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MIN 3141022208
ASSESSMENT OF RESULTS Acceptance Foratest to be considered valid the meanofthe solvent/vehicl control revertant colony numbers for elaabcohrasttroariynusnhloeuslsdoltiheewriwtihsienjutshteif9ie9d%bcyotnhfeidSetnucdeylDiimrietcsotofrt.heThceurhrisattorhiicsatlorriacnaglecownitlroblermaanigneotfaitnheed aatslaearsotlalidngourbelcionrgdoofmevaanmareexviermrtuanmt ocfolfoinvey yneuamrsb.erAslosvoe,rtthheepnoesgiattivievecocnotnrtrooll.compounds must cause Analysis `fTohrethmeeasonlvneunmtb/veerhoicflreevcoenrttraonltgcrooluopnsi.es for all treatment groups will be compared with those obiained Evaluation Ifexposure 10 atest substanceproducesan increase in revertant colony numbersofat least twice the rceovnecrutrarnetntcosloolnvyenctovuenhtiscalteccoonncternotlrs,atwiiotnhs bseolmoewetvhiatdeantcewohifcha tphoesimtaixveimdaolsei-nrcerleaaisoensihsiopbt(aiinncerde)a,seidn two separate experiments, with any bacterial stain ither in the presence or absenceof $9 mix, it will bbee cpoenrsfiodremreedd. toshowevidenceof mutagenic activity in ths est system. No statistical analysis vill If exposure 10 a test substance does not produce an increase in revertant colony numbers in two `csoenpsairdateereexdpetroismheonwtsn,oweivtihdaenncyeboacftmeruitaalgsetnaiicnaicttihveirtyinnththisprteesstenscyesoterma.bsNenocseotatfisSti9camliaxn,aliytswiisllwiblel: be performed. afIfttrhethreesaudldtistoibotnaalinteedstfianlg otuotslaitniesdfyinthtehecrmiuttearitaiofnotreastcplreoacre"dpuorsei,ttihvee"teosrt"dneagtaatmivaey" breesspuobnjseec,teedvetno satnaatliysstiicsaltoprdoecteedrumrienseutsheedswtialtilsbtiecatlhsoisgendiefsiccarnicbeeodfbaynMyahinocnreeafseasl (i1n9r8e9v)eratnadntwiclollousnuyalnluymbbeerasn.alyTshies ofwitvharsitaanticsetifcaolllsoiwgendifbicyanDcuen.netItt'sshtoesutl.d Bbieolnogoitceadltshiagtniifticiasncaeccsehpotualbdlealtwoaycsonbcelucdoensaindeerqeudivaolcoanlg response ifno clear results can be obiained.
DEVIATIONS FROM PROTOCOL Although the protocol indicated that a solvento vehicle would be employed, it was not possible to Tobhteaitneast ssoulbusttiaonnceorwsauss,ptehnesrieofonoref,tahddetdetdsiurbesltyantcoetthheattewsouslydstbeem.coSmipnacteibtlhieswpirtohsetdhuerteesctosmypsltieme.s wstiutdhy.the test guidelines that this study follows, tis deviation does not impact onth integrityofthe
Spagna
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MIN 3141022208
MAINTENANCE OF RECORDS
Arlelmaainwtdhaetp,rospaemrptlyoesftahnde sSppoenismoern.sT(yifpaepsporofpsraimatpel)eaarinsdinsgpefcoimmetnhewhpiecrhfoarrmeaunncseouifttahbilse sbtyudryeawsiolnl
HofunitnistnagbdilointyL,iffoerSlcioenngcetserSmtarnedtaenrtdioOnpearnadtianrgcPhriovciendgurmeasy. be disposedofafter the periods stated in
arAAlcflhritvhseeufrcohrsatmippmel,reistohaednoSdfpsofpniesvcoeyirmeweainrlsslfabnerdocomanltthaeecwtdeaddttaenoadnwhiwilshliabcdevhrietctehaesionSeudtguhbdtyyoDHinurntehtceitnoergdtsoiangnLdsiitfsehpeSoscfaiilennaocrersfeupitohriettrs. rmeatteenrtiiaolns soufbjtehcet omataerrieaalsso.nabIlfefreequbeesitnedg,agHruenetdiwnigtdhotnheLiSfpeonSscoirences will continue to retain the HofutnhteinfgidnoanreLpiofretSicnietnscaerscwhiivleriendteafnintihteelQy.uality Assurance records relevant to this study and copy
RIA,
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MIN 3141022208
RESULTS
"aTrheprreessuelnttseodbtianiTnaebdlewisth1 1P0er1f0.lucTrhooectmaenacsnulvfaolnuyels fqluuootrieddeh(aPvOeSbFe)enancdompeoscitteidve0cotnhteronleacroemstpowuhnodlse number, "The absenceofcolonies on sterility chek plates confirmed the absenceofmicrobial contamination. d"Tehnesittoytaolftchoelocnuylcuorusnotfstohne niuntdriiveindtuaalgaorrgpalnaitsemss.(see Tables) confirmed the viability and high cell "hTehecmureraenntehviesrttoarinctalcocloonntyroclorunatgsefo oftthhee sloalbvoernattorcyo.nrAlpsprwoeprreiawtiethpionsitthieve9c9o%ntcoolnfcihdeemnicceallismi(twsiohf c5o9nmfiirxmiwnhgesreensrietqiuviiryedo)ftihneduccueldturseusbsatanndtiaactliviintcyreoafstehsei$n9emviexr.ian colony numbers with all sins,
FIRST TEST (RANGE-FINDING) N{hoe tseusbtsetranstiaailninfcorlelaosewsiing erxepvoesrutarnettcooPleornfyuonruomobcetrasneosvuelrfocnoynltrfolluooriuden(PwOeSrFe)baitaainnyedcownictenhtarantyioonf incithtpreesernceor abseonfc$e9 mix. No visible thinning ofthe background lawn of nonrevertant cellswasobiained following exposure wtaosP,etrhfelrueofrooroec,tsaenleescutlefdonfoylusfleuoirnitdehe(sPeOcSoFn)d. 6A5.maximum exposure concentration of 5000 kg/plate
SECOND TEST hNeo essubtsetranstiianlsinfcorelaosweisingnerxepveorstuarnettcooPleofniyvnouromobcetrasnsoavuelrfcoonnltrfo]orcioudnets(PwOeSrFe)oabttaainnyedcownictenhtarantyioonf incither the presenceorshieonfc$e9 mix. NtooPevirsfilbclreotohcitnanniensgouffotnhye! bflaucokrgirdeou(nPdOSfFa).v of non evertant clls was obtained following exposure
CONCLUSION sTthoiswecdonnocleuvdieddentchaeto,fumnudteargetnhiectaecsttivciotnydiintthioinssbeacmtpelroiayleds,ystPeemr.luorooctanesulfonyl fucride (POS)
!
090017
16: i --------
MIN 314022208
REFERENCES
AmaMtEaSge,nsBiNt.h MthceCSAaNlmNo,neJl.laanidmaYmmAaMlAiaSnAmKi]c,roEs.om(e19m7u5t)ageMneictihtoydtsesfto.rMdiettaettiionngRecsa.rc3i1n,o$g4e7n-s36a4n,d BGaActTeEriHaOl UmSutEa,tiDo.nG,asRsaOysWLinA:NKDI,RKLLRA,NWDI,LCDOJX.,(PBd,)C.AULKLEANMDSESRu,b-RcDo.mmaintdeFeORoSnTGEuRi,deRl.in(e1s9f9o0r) PMruotceegdeizneisc,iypp.T1e3s-ti6n1g.,CaRmebporritd.gePUanrivterIsirtevyiPserde.s,BCaasmibcriMdagaea.genicty Tess: UKEMS Recommended `LGERGEAETNO,RM,H.ML,., N(I1C9H84O)LSM,utWag.eanndtesRtAinMgEuLs,ingC.o(pE*dsr)e.veHrsainodnbionoEkoscfhMeuritcahgieaniccoiltlyinTeKstILPrBoEcYe,durBeJs,, Second dion p.161-181. Elsevier Science Publishers BY, Amsterdam. ICH (1995) Genotoncity: Guidance on Specific AspofeRecgultatosry Genotoxicty Tests. ICH (1997) Genotoxisity: A Standard BatyofGenotoicityTestingofPharmaceuticals. UMTKWAEEHMAOSTNSS,,ubGD-A.c1oT.m,(m1i9Gt8Re9E)eEAoNnn,aGluMyisHdiesLlio,nfesdMfaIoaDrDMfLruoEtmTaOgmiNec,nroiBbc.i,aTlMesIctoTilCnogHnEyReLpsLosr,sta.LysdPeairnG:t,KIIRI.ORSBKatILiNAstSNiODcNa,l,EDvWJa..lDu.(a5t4ai)no.dn ofMutagenicityTestData, pp26-65. Cambridge University Press, Cambridge. MMdAaRtOioNn,ReDsM..13a,nd17A3-M2E1S5., BN. (1983) Revised methods for he Salmonella mutagenicity test.
{oes
000018
in
TALE Rests inedwith. phimariam TASS:tet 1 rangefnin)
Ma 314022208
|fy dn
sspoomn
c
RoRa
r3n
oposet:triy eck ooops) :+
Foity hock
wT
CRE
FFoossss
Croopmee ++ || @ ||
a"l:s
rroossee
oTonoreogsms ++ || @ || a
Psl I
rroossee
GCoirgggpmmss ++ | @1% | 5
psl ai
E Tiros wo S EC EC
arroossee
prosoinmsosn G-N | IwRN wA I |u|fd
rroossee
owmonrimss |[5 6|w ||oe ||
vroosste
scpoappeesn - | 7a |Jax | la ||
I
7WrCs7 CH EI C EEE E EEEN
[fotos
tube mt[a[wr [ae] we]
[ [eemNni1o 0c%den uilurteieon; pocf s
A
ws
4Scumnitpetoi.ns1 2.0042 olnvcy)
1080s
090019
as:
TABLE2 Results obtainedwith S. phi TASS:m test2
MIN 3141022208
|fom . [=] w [=]+] Pie No.
Adtton
Sm
+ preset
c
she
one Sm ey hk
+
m----------
:
post; sycheck
pps) -
3pforsosee
Goopnirngep 7+ | @
3=
5 3
slpose dros
pgp) supine
+ +
| |
@ 36
3 0
5 7
7lrost
[w=as
apse + | si
"
s
[CTa=TsEE aEE|] [ws ]5]
Jpose Jpose
aComppo)
-|
| 3
[3 |x
||
2 as
pose rose
sop opis)
| |
|
|u| w |
[fms [Ebel
mT [a se[a] Gogpee = wo [sso | | a|
I No10n%dievio:nof C7 WO EC ACE NE
Jon misng
54*BSucnespeptldeenvei.n1,2224 17 olsony coun)
6a LRU
000020
119:
TABLES Real abana with. phim TAIO: es 1 range nding)
MN 31422208
atin
osoinri I
c
1T -H iS esry ck
p=o i
iose cet
[Ep
[POSF
(5000 pg/plate) 125 169 137
3
[POSF
(2500 pg/plate) +
148 130 152
2
B= bl [frpooosseF
y enr v (1750 yg/plate) * I15a3 1la6m4 1ab5e4
-.6
frooss
Creopneorn +2 |lw iwws l|=m
i
ITi-- ros ----1-- T>N --
hpoosw
awnonreem || 3ns
=
p14pr[PoeOSwdF
(no25ppegileante) .- | 8
11=224
row
wipe | m
[[lomwsms Ghamm [ rwswa [woe|r]]
[etmwie
ose 0m ene a]
[None; 10%dilutionof
fra
iSnioparsmam pay ai
22800
090021
20:
TaBLEs Results chad with. phim TAIOH tt2
Na 314022208
LEE [[=1[=] adtion eNeewesmteyry acee rcorsey. cot orosr dpoosstt [OSrwoossee s
rroossee rrooss [[imeetdres [finside (remint cgure, acs
weSparvin | 4 [A 5:
aw osw + |w[mm apie + | [se
T ue |E | 5 me ||
wooesgapenr ++ 1|0i || ns n|ee | u|s|| 2on
E Ton e Tor Toe]
swweipaens orem
| |e |mwmom|| oosw om me ||
ooospen [so {me lus | mow
Gsrpoamin
TmwoIew wwom{mwr 5a] o0[ss |me ar|
.
Ho
4E Sumix tboioisoc .n1,2. e 17olpnyt os)
paid Yay
090022
1
TABLES Resultsobsned with phimariam TASS: st 1 angeTadig)
Na s141022208
EEadtin
dsioonni!
c
TS
fo
1 roosve.trsy heck compen :
riow y row
S Gw p + | =
Cw EEas
drroosse rose
r -- g ||= mono + | 2
nw]I la
rfroose
dra ++ || 7
nbw ul|]
[Toimoe eos
S =r E [wbE[Tw]e] Guoromo | 3s [wae |e]
loross ost
rGeenmay mmfwa=m efw] ts ones | fe fu | w]e
drrooss
sromoonwe ---_ | 3|e [[2as0 ||m |w|e
C IE -- aT 3EO TIT EAEE
E ene 10 ions E = E 0 EE
ersten
tEixcte1 p2.0t02oilloynrn.)
naane
000023
2:
TABLE6
`Results obtainedwith S. typhimurium TAIS3S: test2
MIN 3141022208
Adin
T1 oIeEtrsty rk rsoisst:ycv Spooswe rose ose 6[poSF [owose
fsroilml IN oes
c
coonuppiny :
S somgo + | |
JM
apo + | nw
is
S (1o250opg/plpate) ++T|a 220 e o2ln To [wu1o9 ]
F[PooOsSseE Croi se e [[iTffmmoine e
[ ovremN n 1o 0%cdsn iutlueteiono;fcd
(s2a5o00aupgpplpatse)
-
--
|| 15
|
2[6
|1|=6e | 1o[vnn] 7
cooper J| To6 [5|w {=nnm]
Cr rops) o [3 [om r [aw || ] %|
:
4B Sunt e oon1,p 20017opcco u) _
.J
5: 090022
TABLE? Rests obisnsdwith 5 phir TASSes:t1 (ang odin)
MIN314022208
addin 1 eEe trrsity heck roesey. ce rroos rroossee orossee rose [owFoes rroossee rroossee rroossee [[pweestis [lpAmesnios
eirinets |None; 10% dilution of
Ssonnic | 4
c
o7y
cong ?
ownuas + |x |=
a ao o ++ || 3nn || #7
pTreoswe +SEDR ILu 4|=
proton Sa
+SED [=IF [2%[>]$
sepmuns. | m15 w[0 | |e
V3
aCmopoenos -- | 6|0 ||w || w
;
rposminsaen GR | INH | 5IR ow
3:
Gwe _+T[ [ww i[ha[v5o][a]
Bose | [4 |ws| |
- [|
P*E rap ia1p 2m2 os lln. rs)
50025
a
TABLES Results biainedwith.ophimariam TALS: est
MIN 314022208
Addition
on:55 ry eck 1 onebates check 2fosr,
ry heck f[rroosst sfros efrose feos. [ofSmFwOesmed 7 1ofpos nfeose njeose ]post.
[lowes
[T5[Benzollpyrene
[e[5-Aomniesos1c0riddiimnieonof 17joonvemmiigetncslurs, ped
Somix pret| A st+ s
000g) -
cfm
[Cr asp +
= 1
Ww
song + soppy +
u 2
Copa +
Py
2
Gorgee s w[3 Tw[5 Tw|B [ms]
esoppin wsgpw
- [0 - |8
[4 | a [5 fu
sogpmy Gown)
-o|ons[|unw
TeTw[5s [mw]
Cpe + |55 |as [me |ww[|
Gouge) 126 1376 [3507 |27|3 |
+*SEuxncedpctdladteenvsi.n1, 2.40417 otal olnycous)
arf|
000026
25:
NN314022208
TABLES Results hme with E.colfWeirAJGKVEOL (CBS: est ranging)
Astin T1 oRe triy heck Jrcose vc drroos rroossee rroossee olrose [Tiitoes rose
Ssom|e A
c
Jo2 n
op :
w sons e + |e [me | a|u| Eow (rraprpmon ++ | |e |[o=e unse [fous|msos Coosomoa ++ [[11mm | we ufws ||weo| 0wm arvgpem + |r lm lw | ls S Tm[e ar [wm [me Gomme | [wm | s ||
Eom naan e13|opPoOsSF drroosss [[fmimonies I
[None; 10% dilution of
eran
swoongss (1750 pgiplate)
:--
|1|3wo2|
|
ns
w
nn|wee
fm |w||
9
w
rcoengees |Jw |m| n | |wuw|
ime_+ [Twwm[[a@[[mw y[w ma%]]
-- TE |
4SEaantdptovsi.in1,242 Col sony cous) 20Per sfe) mi
gang
060027
a
TABL1E0 Results bind wth E:cll WezaorApKNIOE CISDs:t
MON 3141022208
woey] non 1Tooe tSrsSty rk r-- os. rroosse dJoosse Thos (m=ee rroossee rroossee [[HEowAert [o(eT meit cr ompas emi
a-- sotme
c
::
[EP
sr np T + | 154
wwo|T=
Waapan +| | 1132
1w0||
T opps o o + | wew m Twe wi| ]an
sroompe +mie ||re ||e |||| on
woaopm ~+ | mm1s Ifmwo vwee ||e||5
Goan IsToww[[Tww w[ew[]|]
osm on[oo [owe[7]
I
4A Satdoin +20 Pay42 rie
2: 000028
MIN314/022208 -
APPENDIXI
Historicalcontroldata
Presentedbelowarethe historical condattafrroomtlhe period 1 April 1997to30 September 2001
Untreatedcontrols
Swain
TAI00
TAISIS WemwApOMIOl TASS (cmso1)
TALS?
Somix
Soe se
TR ee
Minimum
% sm 12 10 0 Bm B 77
Maximum Mean
M9 165 3 09 me 18
29 19
167 21
200 134
48 3
st 4
23 12
3 13
NSou.ndowfrvdaldueevsiation
3196 323
307 3
31 3
24 17
218 0
3131S 45
308 312 35
UpLopweerr9o99%%llimmiitt
151 168 33 76 78 12
29 10
170 76
204 6
49 2
55 28
23 7
35 6
Positivecontrols Stain
Somix MaMixniimmuumm MNeoa.onfvalves Sunduddevision
TAI Cea w@o(m@) (21) $40650 1S36247 1S3610 31260 1175 81889
TALSYS WPurAGKMIOL TAS TALS ee (vT on)eLe eLe 5) @2 8 (26)Im (3Dm3 (3 6 72085 141390 120d B10812 A6S9S 0694 92903 1S01371 139033 5266 314319 41946 71946 418182 41S8 2S0S 7l8S 8l1s0 33679 88100
(ENNGp3g ((E)NENNGN2Gu4Sgg ((8)ASFod-i2u0m0az5iudge 0.505 (8()925N-iAtmoifnloosrc1eind3uie0g ousg 2BeAmniznnolalnptyhrrseancng2eenuge )2-Amincanthra1c0ekneg
gana
000023
128:
Shs riBiatn
MRTay
<&a"%
-
&
HEALY
`THE DEPOA F HER ALTHT OFTM HEGE OVEN RNMT ENT `OFTHEUNITEDKINGDOM
EsBgERLro.urEeEm EH E rO T eEiE= Sn=osrfatmerio.ae, Mutagenicity
2t I D9JEanuarm y2001 aT T Aa T rers
@ Jol
Ber eoffer
rd
3
000030
r. Efquiry umber: 23923D
PCTL T=Tebl
Hunti7 ngdon
Life Sciences
--T-Tbbl.2
PROTOCOL PERFLUOROOCTANESULFONYL FLUORIDE (POSF)
BACTERIAL REVERSE MUTATION TEST
Sponsor SM Centr
3M Corporate Toxicology
BSNutvilrdSaisnigs822.03-222E0-02 Usa
Total number of pages: 12
ResearchLaboratory Huntingdon Lie Sciences Lid.
`Woolley Road
AHCalomcmtoirnnibgguagreoysntive EFNaGLaAhNsD
090031
Pagei
bay 23929
Huntingdon
Life Sciences
PROTOCOL APPROVAL
PERFLUOROOCTANESULFONYL FLUORIDE (POSF) BACTERIAL REVERSE MUTATION TEST
RVi9 nTge% i = Honiton Lite SinesLu
v= U Jer /o(
Flu. Eillatloly
-- ew --
Sponsor
wfofon
ml ------------
PFlic inn btoothScooptlssofhisagri oefor ou recordsand eta an the Sy Diet SvleyboDiornecato bappriovaanefahgededp.roTthocosmpvleettheesilytdtel amg faheeprfaoealnacnesuch dels
000032
.
Enqiymamber, 2930
Huntingdon
Life Sciences
PROTOCOL APPROVAL
T- 662
PERFLUOROOCTANESULFONYL FLUORIDE (POSF)
BACTERIAL REVERSE MUTATION TEST
(Je A
NCH.ianAmlsnggedmAoelnnltLif: Sciences Ld.
3 / 07s /
Date
SrGclomer t. Bulitgy
.
Somer
---31/7/00
FPleaaseisingn LboethScoopineseofthis page, retain one for your records and return one to the Study Director at o SocDirecc toapprovt eofthceep.rTootoocipvoetl oenahdgeeswillybedestdappgeeofrvhehepwrrotoofchneocelischeal
on
090033
Eniynamer. 2350
Huntingdon
STUDY DETAILS PAGE
SSwuadymie:b
sacitrai revre moons
`Test substance
Bch mer: Identity:
B.
ppxppie:rne: Payer Storage conditions: Sub SoSopvefecirtfticbGrsavioty:former:
Analysisof achieved concentration:
viz Perfluorooctanesulfonyl Fluoride (POSF) Claris `Sponsor's responsibility; assumed stable for duration of
study
posi Room temperature NcTooatblessdseeamineidss
`Not assessed in this study
Sponsor's Monitoring Scientist:
John Butenhoff
Head, Departmentof Genetic Toxicology: DrM. Gillian Clare
`Study Director:
Mr Kenneth May
Person actinginthe temporary absence of the Miss Elizabeth Farrall
Locaionofsu: [Ea-- rner `Experimental completion:
DEeppofGeSneut tTo95o3 kT, HuEntingdon Li 20%ovent2e0r1
"5 December 2001
Draft report:
4January 2002
Sens
STUDY DIRECTORAPPROVALOFPROTOCOL
SoKeunnientih Mrayif eee
Lanna
i Ockobes 200
Date
000034
.: Enquiry number: 23923D.
`CONTENTS
Hunti ngdon
Life Sciences
Page
5. MAINTENANCE OFRECORDS ccs
|
6. GOOD LABORATORY PRACTICE wc
Ao
000035
"Str
L
INTRODUCTION
Huntingdon
STyhseteomb.jecTthiosfptrhoicsesdtuurdeyciosmtpoliasessewsistthhethmeuftoalgleonwiicngpogtueindteilailneosf: the test substance in abacterial
Reverse Mutation Test, Guideline 471, EC Commission Directive 2000/32/EC Annex 4D-B.13/14, Mutagenicity ~ Reverse US EPA (1998) Health Effects Test Guidelines. OPPTS 870.5100 Bacterial reverse
Gem gun bro re $5 Fs Japan Ministry ofAgriculture, Forestry andFisheries, (1985) Notification of Director KRYORi No 21 ster 59, JointKDAirNecPtOivAesNofNoJ.EP28A7, IMHW and JIMITI (31 October1997).
i S1990`)pO.lffic,Lialt NottL iceeofJiMOLt8iyFeeebrsusahe rsy 1a9cE 99),yi comeSs5ahSe,,$s1n 7co,awrpossomo,ne mRehere Ey eo en ern e eAmptleochynsiqmuuetabnatsesdtroainnssoifmiElsacrhperriincciples has also been described by Green (1984
s ofEc hia coli which are incapable of synthesising )t.heTahmiisnsoaysctiedm
000036
Emme 25
Hunsnoden
ne
The
ts sd ca
S. typhimurium
ry
stra
final
ins have a
tons
defective
wbhcherenbe tahe mAor SpBrNo1esa.gen cell coat `which allows greater permeability of test
oe anes addition three of nthneom pososstesswatipleaismtiod (fPoKcMh1oTe0t1nl)Emswe.hiyaach.bihvnotrcebodinuiceosrnnanlcer9rsorb-p1rb)oynepeyrpepeawsier
ti 55 or ss + Wh of
2. Soeaicnresnl EXPERIMENTAL PROCEDURE Simei TASS HG fo 73
S. typhimurium TA1537 hisC3076 rfa uvrB.
S. ton typhimurium TAI G6 f TA98 hisD3052 rfar ROU uvrB pKM101
-
rTsE.eiecolotitWnPoaf25trpoculnvriAvapoKncM1h0vi1s,(COMf89o1v)ifNomotheaoCrolllinC=sololfisn olf TipnedCMuatinse
peiesotte eapeartiyndaoesntnerestec.s AelEdxmhBboiogiof pF.oiroTnhies PASO)es
a i mao oe smEasiohn. onytireys of0feIshin 1t0h4
oe a of cdr vil be sed 35 of tn rl
Poinconrise memset ions incubated, with shaking, at 37C for 10 hours. These cultures provide approximately 10 cells
Th ss of i
ESonrietNyn
PS-- isoa
Concentration:
0.5 pg/plate for strains TAIS35 and TA100
Late
090037
CT Enter 72D
none Identity:
r4is 9-Aminoacridine
Hngndon
CAS No.:
-
Solvent:
Concentration:
Tin Identity:
rt pressof9
607-57-8 DMSO
1 pg/plate for strain TA98
ww 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2)
SuConcentration:
a } 2 pglplate for strain TA1535
10 pug/plate for strain CM891
Memity:
CAS No: Solvent:
Concentration:
Benzofapyrene
50-32-8
-
DMSO
5 pg/plate for strains TA1537, TA98 and TA100
Preparation of S9 fraction
S==o . 7
2Sb pr ied He
5 ton il be red romE0Loofllycn 010Jsiinlg,, MYiishedrcitoniocn adxidese `vehicle as a single intraperitoneal injection at a dosage of 500 mg/kg body weight. On the fifth
day after injection, following an overnight starvation, the rats will be killed and their livers
Ten os KO "The following steps will
be
RE) carried
Thout
at
nrfe 0-4C under
ic
dion, conditions.
TTohee
vewri sbe livers will be
$9 mix contains: $9 fraction (10% v/v), MgCla (8 mM), KCI (33 mM), sodium phosphate
..., 2
buffer pH 7.4 (100 mM), glucose-6-phosphate (S mM), NADPH Al the cofactors will be filter-sterilised before use.
(4
mM)
and
NADH
(4
mM).
RE a 000038
vpn ty
St i es
Hungingdon
Ire Sciences
Normal cote oe peta, SAIS in the chosen solvent/vehicle, which will provide afinal concentration of
is the standard limit dose recommended in the regulatory guidelines
is iy los 5000 ug/plate. This
this assay follows.
Aliquots of 0.1 ml of the test solution/suspension will be placed in airtight glass vessels: this
0.1 ml of a 10 hour bacterial culture. The mixtures will be incubated at 37C for 30 minutes
rFoito Cshor earawnlt+bap tis t2en0fshsh hr ct otfo
oo2 rnttke o,oee eog n opneesn3cSsdbibmviialleboehehcoonl,osbi dhreonlnbe Any toxic effects of the test substance will be detected by a substantial reduction in revertant
000039
Eni momber, 259230
Huntingdon
Seconds
IteVfnaiericlriee.oarnopHfonsositpivsivveearcneoasltpaornnasiepmoimasenosbeto5afianncelhdydiinnT)toshesfeirtstnantestct,o3tmh5oecsteo0cfooeandhssttteosteaowsinllnmsbeeasnyexeeacythrepeat of ihtnhigesrogrdpmbresaofnGoogxe.deemoyes19otbebpinndd, theosconnd sspet may betreestded iNihsne tbfe ooenmeee of hecShtypBones oor To es
3. Acepince ASSESSMENT OF RESULTS
CTCanuooomrnblelarscgbfoepner euoiafcshsnsbettrradiwn escshenouryldlaliiesawtittgth+eintStOhsvUe Re9S9%toFcoionfmeidadreneeceovolifrlmitysSvoDofonthre mcursrmsentehiiimstonirsice]al
Analysis
:
"The mean number of revertant colonies for all treatment groups will be compared with those
obtained for the solvent/vehicle control groups.
1VExEposc1o3rtesnt ssteavpreocescamnesweh svomeereadceolnony murbpes Lof adfsat
pnietenssseom.iReo so a5i9mlelso e winnforehon Eon aan er `maximal increase is obtained), in two separate experiments, with any bacterial strain either in
Wf capone 0 st susan dos nt pods a nse in reverams slony numbers in
Sain apse ie two separate experiments,
it will be considered to
pr with any bacterial strain either in
show no evidence of mutagenic
the presence or absenceofS9 activity in this test system.
mix, No
eSC1ohinseyeeenas1ma0 escsnniTanotStioaledtsrismgiyolncinei erdseSreohsdeefmwoicSolnehstE"SppoaresaisoorPe"Wroeiove a sTpYeBe
iwi wy bans of aime elle Domes so Boy en)
Rel conto ea pen foes ne,
'
000040
areneeeeinem gE anLT
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4. ero icon REPORTING dc fo st bsei, eonh.oi. rrekrs rdleorcronns sod their standard deviation. sommes vi ei Where appropriate, graphs will also be included. `Good Laboratory II fSn otheeeabss,ennce oeBe of ongoingFcoopmmunnica)tia onns,dHwunl2ttipnygeodoaentiwLinoilfeenSoecbaiensynhycee]bssehreceosteeo.rnvo+efsodrtmheteasr.igs7htotIron ontoapcrroAmo f ppiplnt iegfSpersir e yeSponse pid n Apps dhloin erofpcs il be slic erro Draft report: 1 od dsl i ih unbound (double sided) rg irs) an sieonesecon1nutnboound 3 arte (single sided) flr- i came 3 3 Grd :
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copy of the final report in its archive indefinitely.
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000041
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6.
GOOD LABORATORY PRACTICE
TSthandsatruddsyasweill bfeorcthonducted in compliance with th principles of Good Laboratory Practice
The UK Good Labortry Practice Relations 1999 (Satory Instrument No 3106.
OENEVCADCICPrHinEcMipOle9s17,of Good Laboratory Practice (as revised in 1997), EC Commision Directive 1999/11/EC of March 19 (Oficial Journal NoL 77)
7. QUALITY ASSURANCE
The following ill be inspected os audited in relation 0 this sty.
Protocol Audi
Jr --
Process based inspections: rReopurteisneentaantdivreepdeitieti,ve pportocneedcuersessawrilylobnetihnispdecyte.d on
Report fue
The drat report and say daa wil be audited before ssc
-
`ofthe draft report to the Sponsor.
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5. HEALTH SAFETY uI1nn97do4er,rdeaarknidfnotgrheHtuhCenonitsnrgoydloonfthLSaiutfbesttShaceniceSenpscoeHnsasztooarrcdsoohmauplslltyoprwHoievtaihldtheheRHeHugneutlaianltgidoaonnsnd 1SL9aiff9ee4t.yS1ca1tonWcocersoknwcitdt.hoAaco1tf informacion avaiable toi regarding known or poten hazards ssocaied with she bling Thaarnldluasesoofcoamnpylsyubwsittahnlcle scuuptpleinedegbiysltahteioSnpoannsorreltoatHiuonntsincgodnocnerLniifnegSschieinmceens.t oTfhseiSrpoennsosr thSeuescthsumibunsfstotarnbmcaeetriceoancniivnbeedthhbeaynfSdoalrfemdtyofianMatnhcaeogmleapmbleoirnaettdoSryeH.ruvnitcAientsgadthoeHnuGnLstiefiernegSddcooinnenLocieffseSSatecfsettteynscueMbssatnsaaengcneesmdehnas: iSenrfvoircmeastioantmaHuynbteinsgcdoenp,ibLlifee Scions, other documenation. somaing. pe. ren
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Dente 2950
Huntingdon
9.
REFERENCES
Lh AMES, B.N., McCANN, J. and YAMASAKI, E. mutagens' with the Salmonella/mammalian micr
(1975)
osome
Methods for
mutagenicity
detecting carcinogens and
test. MutationResearch,
GATEHOUSE, D.G., ROWLAND, (1990) Bacterial mutation assays
LR., WILCOX, P., CALLANDER, R.D. in: KIRKLAND, DJ. (Ed). UKEMS
and FORSTER, Sub-committee
R. on
UGuKidEeMliSneRsecfoormmMeuntadgeedniPcriotcyeduTersetsi,ngp.p.13R-ep6o1r.tC.ambPraritdgIe Urenviivseerds.ityBParseiscs,MCuatmabgreindigcei.ty Tests:
`GREEN, M.H.L. (1984) Mutagen testing using 1rp* reversion in Escherichia coli in: KILBEY, B.J., LEGATOR, M., NICHOLS, W. and RAMEL, C. (Eds.). Handbookof Mutageniciry Test
CTTOSEDe ICH (1997) Genotoxicity: A Standard
MAHON, G.A.T., GREEN, MHL.,
ET ocorhWavsa,teaTsoe BatteryofGenotoxicity Testing of Pharmaceuticals.
MIDDLETON, B., MITCHELL, 1 de G., ROBINSON,
University Press, Cambridge.
MARON, DM. and AMES, B.N.
(1983)
Revised
methods
for the Salmonella
mutagenicity
test.
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