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Attachments to Letter to C. Auer dated May 18, 2000AR) Je Studies and Other Information on Certain 7 Perfluorooctane Sulfonate-Related Compounds 12. N-MeFOSEA | N-methylperfluorooctane sulfonamidoethyl acrylate Genotoxicity 1) TEevsatlu(awtiitohnoinfdethpeenMduetnatgerenpiecatA)c,tNivOiTtyOoXf,T-P5ro8j6e9ctiNnot.he1A1m59e6s5,Sa3lMmoRneeflelrae/nMciecNroo.soTm-e 5869.1 (FMZ-3559, Lot 2408, wide-range), April 20, 1994 2) HEvuamlauantiLoynmofpthohceyAtbeisli(twyitohfTi-n5de8p6e9ndteonItnrdeupceeatC),hrNoOmToOsXo,mePrAobjeercrtatNio.ons11i5n9C4u3l,tu3rMed Reference No. T-5869.2 (FMZ-3559, Lot 2408, wide-range), June 5, 1994 3) MEvuatlautaitoinonToesfttwhietMhuLt5a1ge7n8iYcMAoctuisveitLyyomfTp-h5o8m6a9Cienlalns (IwnitVhitriondMeapmenmdaelnitarnepCeeatl)l,Gene NOTOX, Project No. 115954, wide-range), April 20, 1994 3M Reference No. T-5869.3 (FMZ-3559, Lot 2408, a . 005581 REPORT EVALUATION OF THE MUTAGENIC ACTIVITY OF T5869 IH THE AMES SALMONELLA/WICROSONE TEST (WITH INDEPENDENT REPEAT) NNOOTTOOKX SPurbosjteacntce11356516956 ~ page 1 of 23 - RECEIVED NOV -n9h1494 ToACULOGY 605582 T-5869 - NOTOX Project 115965 - sTaTevenT oF ue coveLIRICE - nnn VOTO 5.4. *s-Hertogenbosch, TheHetherands TThoetesntudeybdeesncriobf"ed fn this report vas conducted 1compl ance with the most The OECD Principles of Goad Laboratory Practice rien are essentially in conforatty with: TRhoe aUntitaednSbtaaterseF?ood and Orug Adwinistration. Tite 21 Code of Federal TFheederUenritendepuSttaatteisonsenypiorroenm6e0n:tal Protection Agency (FIFRA). Title 40 Code of TFehceerUantiteRdeSttaatiens sEnvpiarrsonmoeonp:tal Protection Agency (TSCA). Title 40 Code of riothrt:he exception that the stability of the test substance fn the vehicle fs stu otrector Tog. .3. van de vaart em Date: Yr pez 005583 T5869 - NOTOX Project 115965 QUALITY ASSURANCE STATEMENT _------ NOTOX B.V., 's-Hertogenbosch, The Netherlands. sSpteucdiyfipcropcreodcueressseswewreeresubaljseoctintsopepcetreioddiact pienrsipoedcitcionisntearnvdalgse.neral non study Tanhdis rreespuolrtts awacscuraautdeiltyedrbeyfletchte NtOhTeOXrawQudaaltiat.y Assurance Unit and the methods DATES OFAUQDAIUT_SINSPECTIONS/ REPORTING DATES. 24-01-1994 26-01-1994 15-03-1994 24-01-1994 26-01-1994 15-03-1994 Quality Assurance Manager C.J. Mitchell B.Sc. Date: 20 ~L- At - page 3 - 0605584 T-s86 _REPO--RT A-- PPROVm AL STUDY DIRECTOR: NOTOX Project 1196s -- Ing. E.3. van de aart Y= vate: sp/ou 9 MANAGEMENT: TOer.chnIiCc.al EDninriengastor vate: 20] ou liggu -- 605585 T-5869 NOTOX Project 115965 -- PREFACE _-- Sponsor Study Monitor Testing Facility Study Director Technical Coordinator Study Plan 3CManaBdealsgtiruanat- 1Ch1emical EBC 8Be-l2g0i7u0nZWIINDRECHT Mr. R.H. Cox NHOaTnObXakBe.nVw.etering 3 5Th2e31NDeDthe'rsl-aHnedrstogenbosch Ing. E.3. van de Waart G. van Cort. CSotmpalertetd:: JFaenburauraryy250,4, 11999494 TEST SUBSTANCE _-- Identification Description Batch Purity Specific Gravity Itenssttrsuucbtsiotnasnceforstorage coSntdaibtiilointsy,under storage Expiry date S4tahbolures foirn vaethilcelaest T-5869 Crean solid 2408 95% 15 At room temperature fn the dark Stable January 01, 1996 Oimethylsulphoxide: not indicated VEHICLE -_-- Tqprhuieaolritteystto (usMseuerb.cskt)a.nceTeswtassusbusstpaenncdeedcoinncednitmreatthiyolnssulpwheorexipdreepoafresdpedcitrreocstcloypic - page 5 005586 T5869 NOTOX Project 115365 e cutoELINESmt----m-- m -- e -- e TShueidesttuidnygs.procedures described in this report were based on the folloring - OGTorurgiiadcneoilTsioangteyis:onfoSrafolrsToensEetcliolnnagosotfyfcphCCihome-muoirpcieaurlnast,iRoevnGeuriasdneedlMiDnueetvaetlnioo.opnme4A7n1ts:s(a-OyEG.CeDn)s(,taidsoOpEtCeOd - EJ EEuCloDpieraenctEicvoer-- neos6t7c/S4C8o/mEmEuCn,ityPar(tEEC6)1,HoDtihroedcstivTeor92t/h6e9/DEeEtCe.raAtmmaemxionV oobf the 1RT5eo5vx2ie)cr:istey:utBa.t1i4o:n"AOstshaeyr".EffEEeCctPs-uHbultiacgaetniiocnitnyo:. LSe3l0a5onTeeddivas.EteysahDirsmuesrbeerm, - A-- ReCHrIVING --e-- rr ---- NPrOoTtOoXca8i.,1. rweip1o1rt,arctehsitv.eartthieclfeollroewfienrgencdeatasafmorpastndTearsatn 1u0thy.ears: oaaerIve Purpose of the stusy Tt0hyeshiionndbuuerccietuinrveebvaecrotsfeertimhauilstatssittrouandisynsyianstoatopgreoendvsuacloeufathhieisstttihidediinnteee.s-trienqsduueibprseintdnaegnnctesSafErororansTiintisssoafbility preRsSeenRcTeTSo-foragRaentyaobnost.icTsneysatsesaay.(Sb9e-sm.1xb)o.ndurtes nthe shoo: amy Sone Justification for selection of the test systen aTThuheteagAbeanncetsiecrtiaeasclttivshtiartsayibnoesfeneamsphlioodwenyedrtoawnegbreeeoacfarpcaahpbeilmdiecaaonlfd CIonmepxopuenndssi:ve indicator for Getbcting both frameshift the and Base-pair substitution autagens. page 6 - 005587 T-5869 - MATERIALS AND METHODS NOTOX Project 115965 _ TEST SYSTEM _--_-- Test System Rationale Source Salmonella typhinuriua bacteria Rreeccoogmnniezneddedbytestthesyisntteemrna(te.igo.nalEPAg,uiOdEeClDi,nesEECa)s. the OBre.rkeBlreuyc,e and TAL00; NU..S.AAn.es,(TUALn5i3v5e,rsiTtAyI53o7f 1993) aCnadliTfAoSr8n;ia1a5t87 The characteristics of the individual strains were as follows: strain Histidine mutation Mutation type TTAALS5B37 hRiiss0c33005726/R-factor* FFrraanneeshsihffitt TATAIL0S035 hRIiSsGG4i66/R-factor BBaassee--ppaaiirr ssuubbssttiittuuttiioonnss *: R-factor = plasaid pKMIOL (increases error-prone ONA repair) Each tester strain contained the following additional mutations: gEIaflaT: dmRuietkeaapttiirooonnughiinn(ntdihetefreagctateliavcerteo1dsiaeptoempseostlaybsaoclcihsamride cellcoat) BVlros:: dJoesfsecotfivethebioetxicnisisoynnthreespiasir syste (deletion of the ultraviolet. repair 8 gene) S(-t1o9c6kCc)u.ltSutrreasinosf wtehreeforuergulsatrrlayinschewcekreedsttoorecdonfiinralitqhueiidrnihtirsotgideinneTrAeLq0u0i)r,eaeUnVt-,sencsriysttiavlityvioalnedtthseensniutaibveirtyo,f aspmopnitcainleloiunsrreesviesrttaannctes.(TAS and CELL CULTURE ---------------------------------------------------- Pbarcetpearriaatlionculotfures Samples of frozen stock cultures of bacteria - w(e0rxe01dtrnaon.sf2e)rreadndiintnocubeantreidcheidn anutsrhiaeknitngbbraotthh 0(.307.C,Of105.04spaat),700untniml (t1h0e cceullltsu/rmels).reFarcehsehdlyan gtersotw.n cultures of each strain ere used for a Tpage 7 - 005588 T-5869 - NOTOX Project 115965 - Agar plates Top agar Environmental conditions AaTiggtaaerrrspRelda"1it8uensg.p(uGsrliu9fciocesa)de caaggoaanrrtaim(ne0eaxdoi1uan2,5cocao1ndteag.ilnu1ec30od)sepeirn V5(go%)gebViooBgtoeinlnn,era0n.d0M.0e.d6ainuamggEhai(ms1et)r,i,di10noeH. .lo1c95o6s,. 005 pAucreitfyilcoartni]otnhinaandsesoomfepEsrchoerpischriaiec3o.st.:Hopra.rtial Chen. 218, 97-106. VpSoaugmrepi?lfeiBseodnonfaegra3raNveadsttuospheaatgaecrdonwtteaoriedniintsgrsaon0ts.vte6eXrtrh(eewd/va)gnaero. 1t0ub8e1s VGelraessauttuobcestavweidthfnoreta2)0 Bcaipns.atTo13p00aLg,ar aAt1ht1e3e7ixnCpc.eurbTanhteeinotnt.semwpeerreatucraerriweads soounti tinoretdhecdutarnkg - REFERENCE SuasTAICES VNTRegeathiveTceonatTfroelt:he test article. Positive controls: VS TitAhoHuStran int metae bsooldiicut naazcitdietve ato(iswo)nr C(F(-lsuE9k-t a1x))L: opiTaotg e Ssoatvnenet TASS THs SG{aaaumanniosnnsoyeaencirnnieidim(nOieW))(9(4S6t)gsa) 4g sane fou sane TAI methyTasthanssuironate. 55) 0 ug ow (Merck) t TiAtDhSmSr,ametaboleitc eacrtI ivation (9-10): attopiate solvent weTA,ISH7 2-aninoanthracene (24%) 5 wg ouo THi0O (2s-iagsst)noanthracene (284) osu owo SpDoMlSrvOeSarliynie t+=ssGfioTzretorheyfTesruoelnpcgheosxtsaiuTdbiaesnteaao.nfc(lesWsepaetcttarlosPchoaprieca quNaeldi.ty"8.1(.H)are). "pages - 005589 T5869 NOTOX Project 115965 - mMETr--ABOrL--IC--ACT--IVA--TIO--N S--YST--EM ------------------------------ Preparation of 59-homogenate: RoratSpMrvaegruemDiacrwolseoymarlatse,nzywmhiecshwweerreeroobuttaiinneeldy pFrroempaBrReLd, fSrovmitasdeurlltndm.ale Wistar TTihanebtoraranaptteearartlyosncewoaenlrdlieytihowoniusts,hedaasatsdoelNsuOctTriOioXnbeidn(20ia%nswtp/hevec)taSlOOfPrsAo.roomcTihuaenrderr1at3s5s4twaen(5rd0ea0r.dimnajjeicgtedbody Ws(aEtcIhrGeiNyfti)wceeri)en. CdToehrnenedo11i1av.ecrcseFsisvoeftodthaeyfsorodaTtastfeorwr,eraettheTryeeamsowtveerde12kahiso1eu1pretsdicbpryrieycd.eedcaianpnidgtewbaisohnesd in cNi0na1'-d3EDv(To0Al.Cu)meSsusbtseoefrqtupieehnots0ip.yh1atHtehesboudlffifuvenerrspihowteshrpehaamtiePnoctbetudefrfeirhnoma(opgbhelne7i.nz4de)arr.eoaTnnhdteaihonhmioonsggaenmoir1seesdmt : vianstocesntterriilfeugeamdpufloers,15whAiinchatver5e00s0tog.redTheinsuTp1earuniadtannitt.ro(g65e)n w(a2s.96teracnyeiornce Preparation of s9-ntx: S$99--aniixx 5.5 ul awcqaousnatapbirinedepedasrtpe;edr2i1m0umlemd10i:.a5te3Hl0ynsgobdeiNfuAomDrPephuaonsdsep1ha5an.dt2ekbeHup)ftfgelornocpoiHscee7-.6d4-u;prhion3gcpohhtahc0ee.0t8eisnt. HfsEroCa1,cutfiioosnno,slutwwiheoirnec;hni1axsem1d a0adnd.ed3d3Fa1f1ttKeCeIrr fs(io0il.tu2et2rioysnat,je-rsaintldairz0ia.it5jizoaen1d 5(9a.parTth.efaoboovethe of the S9-nix 59- components). -- EXPeER-- IeMENT-- AL PrR-- OCEr DU-- RE -------------------- Selection of dose levels SwtieetslhteecdsttiroiannintorfiTApaLln0iDca,adteebqouta(httehiwsirtahpnrgeeanldionfiwnidatorhsyeosuttweSas5st-mwbiaaxss.edrEeipognohrttaedpcoranescleinamtiprnaaatrrityonotsfesttwneere tFeisrtstaretxipcelrefaeunstedofintthheeasuutbasgeeqnueesnits masustaayg)e.nesTihse ahsisgahyestwasco5ncemng/tpriaattieo.n of "pages 005590 T5869 NOTOX Project 115965 Direct plate incorporation assay *: Fstieuvsbetsetdadnicbfeoftehrweenrwteithdteossatenesddw(iiwtnihtohturtiapp$lp9ir-coaaxitixematiiennlyceaacchhhalsfst-trlraaoiignn,.steTiphnse)ttwoeofstitnshdeuebpstetensadtnecnet was effxropeleslrhoiwmibenangcttses.roilaTulotpiocnuaslgtaurwrereien(1st0uo9gpcceagesalsrlisv/etmlulyb)esadodwfeadsonemtoeloft3edmt1heantdotpeshteaegaratre:dstr0ta.oi1n4s5m,lC.0o.f1Them1 ocnfaosnea-acdoftiilvauactttiiioovnnatoiafossntahyaess)st.aeysstT)hesourbisnt0ga.r5necdemi1enft0ns.1ONwSeHDr,ephomasinpxdehdaetieotnhebrauf0fV.eo5rrtem(1xinaSnc9da-smeitxheof(in dscaoornlktiedniattfico3fa7tCitohneforotfop48tahehg.artoApfttuebaregarwt,ahsisthpepoeurprileoaddteosnrteovweerrateasntetulrenc(ethdiivsetainddaignaierncpuilbnaadtteeep.dendAiefnnttet)rhe colonies were counted. *) cAnaersc,inoBg.eN.n,s WacnCdannnu,tagJ.ensanwdiYtahmatshaekiS,alEs.o,ne1l9l7a5/,saMneatlhtoadns mutagenicity test, Mutation Res., 31, 347-364. mfoircrodseotneecting *) Mnaurtoang,niDc.Hi.tyantdestA,nesM,utaB.tNi.o,n R1e9s8.3., 1R1e3v,is1e7d3-m2e1t5h.ods for tne Salmonella Erratus, 1963, Mutation Res., 113, 533. Colony counting WTihtehraenveArrttaenkt mcoodleolnfe8s80 (choilsotniydicnoeunitnedrepeonrdemnatn)ualwleyr,e ciofunlteesds atuhtaonma4t0ically pcroelcoinpiietsatpeer toplaitneterwfeerreeprweistehnta.utPolraatteesd manually. wciotlhonysucfofuinctiienngt wteesrte acrotuinctleed ACCEPTABILITY OF ASSAY aA)n ATnheesnetgeasttivies ccoonntsriodleredadtaacc(enpustbaebrleofifspiotntmaeneetosustherevfeorltlaonwtisngpecrritpelraitae:) sfhorouledachretaessotnearblystrbaeinw.ithin the laboratory backgroundhistor ical range b) Tshteraipnossiwtihviechcaolntsroolrecahseoaniabclaylsarsehouwiltdhipnrodtuhecelarbeosrpaotnosreys hiinstalolrictaelster Fruarntgheerdnoocruem,enttehde mfeoarnepalchateposciotuinvteschoonutlrdolbesuabtstlaenacset. two times the ) Tcohnecusrerleenctt.edvedhoiscelercaonngetroslhogulrdoupincmleaund.e a clearly demonstrated by the preliminary test with strain TtAoLx0iOc coronscheonutlrdatieoxntenads to 5 ng/plate. page 10 - 005591 T-5869 NOTOX Project 115965 ~~ DATA EVALUATION AND STATISTICAL PROCEDURES -_-- ANotefsotrmaslubhsytpaontcheesiisscotnessitdienrgedwasnegdoantei.ve ir: (not mutagenic) in the Ames test a) Tishenottotaglreantuemrberthaonf trweovetritamnetssthien ithout metabolic activation. asnoylvetnetstecrontsrtorlainvalaute,anywicthoncorentration b) Tihnedenpeegnadetnitvleyrreesppeoantseedsheoxupelrdimbeentr.eproducible in at least one aA)tIertsetveisrnutdaubncstetssanwcaietthilsearscetosnpseaic2dt-efrtoeolddt,pheosdnioutsmiebveerrel(aiutntedadugceeindniccbr)yeiantsheethfsneolAtvmheeesntnutmceobsnettrriofol:f in aHnoywevoefr,theanytemsetaenr sptlraatiensc,ounetitohefrlweistshthoranw2i0thoiustcomnestiadbeorleidc significant. If the test substance showed in the first test tacotibveatniootn. only a pthioetshitfiidvrosestesrteejssuptso.tnsebelaotwoanendorexctweoedicnogncetnhtorsaetisohnosw,ingtheposaistsiavyewaesffercetpseatiend b) Tihnedeppeonsdietnitvleyrreesppeoantseedshexopuelrdimbeentr.eproducible in at Teast one eTnhteerpreicnetodintghecrFiintaelriaevwaelrueatinoont adebcsiosliuotne. and other modifying factors might - page 11 - 005592 1-586 NOTOK Project 115965 ~ ResuLTS e-- TOKe ICI-- TY OF--THE -- TEST S-- UBSTA-- NCE ---------------- TSofohlevceosnlutorvniicveoasnltornoofthptelhaecpelTsaA:tLeOsD ccounlttauirneinwgastdheetetrensitnesdubsbtyanccoempvairtihngthtohsee.nounsbetrne vBuagos/tphlnoaIttn.e rthe(edTuacbaelbdesenU1)cp.etBoaansdaenddproeInsnecnlthcueedsienogfcatSte3as-,taixstuhbetshtetaenssctu.er.vsicuvobanslttaennoctferaswttairsaa.nisnecoTtfAeId0SO00up0 p5res2enndceincolfud$i3-n3g1xa. concentration of 5003 u/plate Ih the sssemce and es TmHE A-- MES S-- ALMON-- ELLA/-- MICRO-- SONE -- PLATE--TEST-------------- weTinattbhilrees7.3d18o6sa5en.drAa2nlgsehbaawocfttehtrehiearlteesssuttltrssauibnossftastnhhcoeew,eSdatisncoe.ngeanltoilvaed/oasirece.drrpoeesnroaanstese.dp.olvaettrewo-tfneeosltd, eSixnctprheeiranismeea.nutrisn.latbhTehoernautnmoebrgeyartibvioecfkgarrneodvuensrdttarhnaitissnt-osinrp'iecctauiol?.icirannpdgoeespsi.etnidIvennedtisccyaanttireneegpnattvhaaagjtueisnevere fFeasntctCioonnedditipornosperwleyr:e optinal and that the metabolic activation systen - concLuston nnn B1a5se1d0tonmuttahgeenriec.sulitnstohef Atnheiss sStauTdayuneiltlaf/saiccornocsloundeedastshaayt. the test substance page 12 - 605593 T-5869 NOTOX Project 115965 -- TABLE 1 MTUESTTA!GENIC RESPONSE OF T-5869 IN THE AMES SALMONELLA/MICROSOME PLATE Experinen1t ----e--e -------------------------- D(ousge/prate) M3SetarrnaeipnlnsiucaabofteerSop.lftaytprehesivseur(rt+ia5un.nt0.()Hiwsit)thcdoilfofnfeerse/nt eeet--------T--AS-- S ---- TAS--S ----Tas-- s ----A1--00 ------ Without 9-mix 103 10303 1030303 5300000 SPooslivteinvte ccoonnttrrooll! 67::43 58:43 5:1 26751:122 55:i22 86::35 7:4 3569:2239 1163.i31 119i:36 18:4 6w21.3e320 11306 :+ 214 120::52 1d2i:a6 113m:1:0 113732:3219 103 10303 1030303 53030000 SPooslivteinvteccoonnttrroo1lt ith 59-ntx 58::11 589:::122 2480::38 56.x24 67::32 541 50774.118 211.:52 1166::55 16:2 414161.328 1133s+ 129 11886s352 1158::85 11530:51108 61001s.n31 *10T.e1st,msubosutsaonce precipitated slightly on the plates - page 13 - 005594 7-586 NOTOX Project 115965 Z TABLE 2 MTEUSTTA.GENIC RESPONSE OF T-5869 IN THE AMES SALMONELLA/MICROSOME PLATE Experinent 2 (Duogs/eplate) 3Mearneplnuincbaetre polfatreesver(txan5t.0.()Hisw*i)thcodliofnfieerse'nt Strains of S.typhisuriua TALSIS TAIS37 TASS TAL00 313030 313030000 5000 sPooslivteinvteccoonnttrrooll: 5Bs:14 1100:% 44 8:2 2746:+228 193s:51 180::32 10:3 5777++839 1189:443 2292502s7 29:3 62431+:528 15191::204 1165834s1174 129% 9 111640344388 313030 130303000 50002 sPooslivteinvteccoonnttrrooll: 1724:15 98::43 751 217844324 With $9-mix 59::24 225::43 57::32 2266::25 7:3 Bel 6685:.48 22535+.37 11326841i3 113346:1147 17:5 14909:$1080 0T.e1stmsub0MsStOance precipitated slightly on the plates page 14 - 005595 T-5869 NOTOX Project 115965 = APPENDIX 1 tEhveidefnocleloowfingtesptrecairptiitcalteiopnreccoidpei.tate on the plates 1s recorded by addition of r PeREeCIP-- ITATI-- ON ceove-- e -------------------- oereefin--itio--n ------c--hara--cter--isti--cs ------------------ Slight precipitate DHcoiowusentvtieinrng,guistohfheedthpebryepcinapotiteti.acteeabldeocsprneocti.piItnaftleuenocnetshuetoprlaattee.d Moderate precipitate Heavy Precipitate pDDliiassttteii,nngguruieisqahuneeiddrinbbgyy taahemlaarprlkgaeetdesaaomtuoonutnbteofohfanpdrpercceiocpuintpttieetsta:steaonnthtehe ee e er-- Plate-- , makin-- g the -- required-- hand c-- ount g1-- PFIeerTee-- .-- - page 15 - 005596 T-5869 NOTOX Project 115965 - IAndPiPviEduNaDl IpXlate2counts; (following pages) sEtxrpaeirniaent Tass1 vITHOUT 59-400 pate 12 3 wm wm ooseug/prate) 15103000 n0a 0asi s33 6I51sE3 S5a35o0: LPE 72 HsHii SPooslivteinvtecosnotnrtorlo1 7826sgmae7.i2i VITH 9m Mate 12 3 mew ow beseGuarpiate) 11050303 F 5ios0I s 67 5sHiE1 33000000 sSoua sog ssiit sPoolsvietnitvecconotnrtartol 3s1o2uuss a05}. 8 Test substance precipitated stiantly on the ates RE 005597 71-5869 sExtpreariinnent TA15371 NOTOX Project 115965 WITHOUT $9-MIX plate 1 2 3 Dose(ug/plate) 313030 65 5473 13030300: 27 53 13 5000+ ous SPoolsvietnitvecconotnrtorlol 3264 3706 3809 MEAN SD 55a:22 86::53 7% 3596 +3293 WITH $9-MIX plate 12 3 Dose(ug/plate) 130303 59 24 67 133030004 37075 98 50004 565 SPooslivteinvtecocnotnrtorlol 4868 5136 5216 * Test substance precipitated slightly on MEAN SD 56.x24 576::5123 5077s+181 the pates - page 17- 05598 T-5869 sExtpreariinment To81 NOTOX Project 115965 ~~ WITHOUT $9-MIX plate 1 2 3 Dose(ug/plate) 130303 7 B1B2 1132 130303004 2u6 u16 o15n 5000+ 5 16 2 SPooslivteinvtecconotnrtorlol 63178 58169 63195 MEAN SD 6B:ol3 4vsie3 Bs 4 6o1 e+ 302 WITH $9-MIX plate 1.2 3 MEAN SD Dose(ug/plate) 130303 2u0 2o62 210 21a8:. 52 10330300: 222 117 11 66:: 535 50008 8 16 15 6: 2 SPooslivteinvtecconotnrtorlol 4875 41193 4210 a116:: 328 + Test substance precipitated slightly on the plates - page 18 - 005599 T-5869 NOTOX Project 115965 - sExtpreariinnent A1001 WITHOUT $9-MIX plate 1 2 3 Dose(ug/plate) 130 11203 113388 115012 10303 221112B7L w16r7 333 1000 11662 11252 113326 353030004+ 113338 115333 11248 SPoolsvietnitvecocnotnrtorlol 1318530 1136389 1139750 MEAN SD 1u3o6 +142 2102:254 1239:s6a #833:+150 1317623 21 1219 WITH $9-MIX plate 1 2 3 MEAN SD Dose(ug/plate) 103 1020 112873 112345 13330 +2129 13030 11337 113282 u1s3 3u6s ++ 25 1030303 114328 115401 112561 u1s0:i8s 53030300:0 113259 116282 115835 11530 111140 SPooslivteinvtecconotnrtorlol 613672 518862 156861 7001 +11311 + Test substance precipitated slightly on the plates page 19 - 005800 7-586 sExtpreariinnent TA15352 NOTOX Project 115965 WITHOUT $9-MIX plate 1 2 3 Dose(ug/plate) 313030 B6 s6 46 103303004 57 1135 18 50008 6 8 10 PSooslivteinvteccoontnrtorlol 29%8 2834 227 MEAN SD 85+41a 00::44 8:2 2746 ++282 WITH $9-MIX plate 1 2. 3 MEAN SD Dose (ug/plate) 130303 89 108 1&8 27:451 130303004 0 E5 TT1 98:i3a 5000: 857 751 SPoolsvietnitveccoonnttrrooll 1859 2175 2538 21874+324 * Test substance precipitated slightly on the plates - page 20 - 005601 T-5869 Exspterariinment Th15372 NOTOX Project 115965 - WITHOUT $9-MIX pate 1 2 3 Dose(ug/plate) 1300 103303004 5000 SPooslivteinvtecocnotnrtorlol B8 U8 1o0s ssz2 791 8 13 8 6795s 51u6 s53 ean so 1ER 3s 08::23 0:3 5777.Le3s WITH $9-MIX pate 1 2. 3 ean so Dose(ug/plate) 130303 0us2 37 59:.42 130330004 50000 83 903 38 78s 57::32 7:3 PSooslivteinvteccoonnttrrooll 53 76u0n 55110 6658s= 864 + Test substance precipitated Tightly on the plates - page 21 - 005602 T-5869 sEtxrpaeirninent Tse2 NOTOX Project 115965 - WITHOUT 9-1 pate 1 2 3 wean so Dose(ug/plate) 1301500300 81 18 222B21 0F9E:.23 3S30500:: 33%anon 3Bi:3 ; Psooslivteinvteccoonnttrrooll 62 so1b9 eaat en2s .is2i wT 9.x pate 1 2 3 ) Dose(ug/plate) 1303100300 2mvon3 oo2ma 2=% i:442 E33s%: 2 2aoa 2FE% is SPootsvietnitvecconotnrtorlol 0nw 226 42 2239 173 Test substance precipitated sTightly an the prates page 22 - 605603 T-5869 NOTOX Project 115965 - sEtxpreariinment TAL002 WITHOUT $9-MIX plate 1 2 3 Dose(ug/plate) MEAN SD 313030 116605 115954 116535 5m9os+ 24 1000 33300 116502 115678 114808 6583:+ 117 50004 139 121 127 29: 9 Psooslivteinvteccoonnttrrooll 1105254 1111586 1218760 1114630 3+ 388 WITH $9-MIX plate 1 2 3 MEAN SD Dose(ug/plate) 313030 11215 113229 115114 113268:+ 1143 130303004 u11e 113s8 114238 B13s6 L+ 147 50000 1 1:2 137 Bs PSooslivteinvtecocnotnrtoroll 319328 51981 143828 419309 ++ 1008 + Test substance precipitated slightly on the plates - page 23 - 005604