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ARR2G 328 467 ARR26 _ 328) --_ oDumronizes66 TRADE SECRET Study Title H-25777: Inhalation Approximate Lethal Concentration (ALC) in Rats AUTHOR: David P. Kelly, B.S. STUDY COMPLETED ON:July 8, 2003 3 PERFORMING LABORATORY: E.L du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 ~~ LABORATORY PROJECT ID: DuPont-12966 p-- `WORK Raauesy Novos JA ~ - TTT `Company Sanitized. Do note conts ain TSCA CBI BHETTiisn pi oeLmCOmormonmtiRs oapenas CERTIFICATION `We, the undersigned, declare that this `report provides an accurate evaluationofdata obtained from this study. TE `AnatomicPathology le fSECmTfRrmets for ety Sty itr:Lindy 3300 day.0w; ~ _ Com nA E22957777:7 nbaattiioonnAAprpormoavse LoattoCCoonnennioonn ALLCO)MiRn Sass STUDY INFORMATION Svnonyms/Codes a) * H-25777 Submitters Notebook ee Haskell Number: 25777 puf ponrirases Phusical Chases: Pale yellow sid Stability: dTcbohsneedritvetseitdosnubostfahencesatppdeanroedevtiodbeencsetaobflemuendteratlhewas Sponsor: EL du Pot de Nemours and Company Wilmington, Delaware 19898 USA. Sy Iniated Completed May , 2003 se report over page) rr p------------ -_-- L1E25T77T7::InInhhaallaatiioonnAApppprrooxxiimmaatteeLLeetthhaall CCoonnccenevnartioonn((AALLCC))iinnRRaattss_ DuDuPpoonntt1122906666. STUDY PERSONNEL Study Director: David P. Kelly, B.S. PrimaryTechnician: William E. Ellis, Jr. Management: Arthur J. O'Neill, B.S. Anatomic Pathologist: Management: PSetteevrenC.R.MaFnrnam,eD,.DV..VM..M,.D,iPphl.oDm.a,tDeiAp.lCo.mVa.tPe.A.C.V.P. `Toxicology Report Preparation: Maryanne M. Wilford, B.A. Management: Nancy S. Selzer, M.S. Laboratory Veterinarian: Thomas W. Mayer, D.V.M., Diplomate A.C.L.AM. --_ --_ BompanySanitized.DoesTotcontain TSCA cay - J5:25777: InhalatoionnAApppprrooxxiimmaatteeLLeebtheallCCoonncceenntrrattiioonn((AALLCC))iinnRRaatsts_ DuDuPpoonntt1122996665 SUMMARY apTecwroionoddgertnoosuaHpt-si2oo5n67fa7em7roaislnoelaiCri.rn ltT:heCesDtt*eas(ttSmcDoh)saIpmGhbeSerrBe.sRTwrheatresecwgheearnmeebreeaxrtpecodosnbecydenhnteorasateti-inoognntlshyeofftoerHs-at 2ss5ui7bngs7lt7ea,nw4cee-rhetooufrorm m1-eaosru1r4e-ddagyrraevicmoevterircyaplelryi.odR.ats were weighed and observed for clinical signsoftoxicity during a FmRoaaltslsswoemwierndegieaenxpxaoesruporedt,yoona3ratammstioscspdfehireaormmdeetstehoresf0f.oH1r-0t2hm5eg7/7a7Lereaotxsopmloesatuenrsetceowdnecwreeenrtrerea5tt.ai8ionanensdodffo6r.0a8.1u10m-,odrary1er.s8peecmcogtvi/evLre.yly.The pepeexrrpiiooosddu,;rtaehlewraeetrsemafreritonamiintneh3gde fa0o.tr1sa0wm1eg4r-/edLareygtrraoeiuncepodvweferroyearpesra1ic4or-did.faiycAretedctfohovereaernnyadptoeofrmtiihocedpi.aRtrahetossplfeorcgtoyimevvetarhleuecaot1vi.eo8rnym,.g/AlLl ta0tsH-f2r5o7m7t7hoerd1.u8rmign/gtLheerxpeocsouvreerywpeerreisoadc.ificed and discarded.Norats died following exposure OOctfohnHcee-rn2tt5rh7aa7tni7o,snl.tihgehHrtibesotwodelyroegwiencioaglshietgxnaliomfsiiscneaasnttmiocelnionaifcrasaltsuisingernx3speooofsbd6esderrtavose0do.ni1ne0dtmhagey/eaLxfptsoehsroeewdxerdpatonssoauttrreteeoiatth1.m8eermntgto./oL. related effects. ~ g`rUenadteerrtthheacno1n.d8itmigo/nLs.oftAhcicsorsdtiundyg,ttohHeaaspkperlolxLiambaotreatleotrhyaltocxoinccietnytcrlaatsisoifni(caAtLioCn)s,fHor-2H5.7275777i7s is 6mcao8nsspsimdm.eerdeTidaartngoeabtceerdaotdMymMnoasAmtDiscSlidgiihnatmlieynthteaorlra(tiiMo(nMaeAsrtoDusd)oiloefsAtaLhrCeef0tr.eo8s.tme2d.10a1m0e.gr4/oLps)maoiltnotmhaaillslecoowrnatmcsea,nxtHrioamwtuieovmnelrwu,ansgthe exposure. Had the toxicity could have gbeeneenrastoemdeawehraostolgrbeaeteenr.smaller at this concentration, i is possible that th _ `Company Sanitized. Does not containTSCACB! - H:2577; Inhalatinon AopproosinmatelLeothbalCCoonnceenntraaitioonn (AALCL) iOn RaRis bDuuppoonei122996666. INTRODUCTION tT(ehAseLteCod)bwjoehfcHitc-ih2ve5cao7fut7sh7eidsintshmteaudldeeyawtrahatossf. tooTnhdeeetoAerrLmmCionriees adeex4fp-ihonoseuedrdaisrnahttahleeailttoihowenersaotpnpartthomexosidpmahayetroeiflecextcphoaonlsoceuonrntecreaontrtiwroiantion aetxlpeeacstted14roduatyeos ffoploltoewntiinaglehxpuomsaunree.xpTohseurienhaanldatiwoansrroeuqtueoesfteexdpboysutrheeswpaosnscorh.osen based on thtehin STUDY DESIGN `2Tw:we1ro0emgegrx/opLuopessxeopdfonso6usmrea-elowneelrryearfseotcaraaicsnhienwdgeflero,er4ae-x1hp-oodusraeypdreerticoooadvr.eorsFyoopllelraoitwomidon;sgtphheeexprroeessmourafei,nHi-3n2ga5t37sr7af7trsiownmeatrih.eerReattas roercoave1r4y-dpaeryiroed.covery period. Rats from the 1.8 mg/L exposure were reained for a 14-day ined Rexaptosswureer.e approximately weeks old and weighed betwen 258 and 279 grams a th time of --_ ah`enansdiecmloailbnssiectarelvresedigocnbassocehfrtdvoaexydicffiootrrymmoiormrtmtaleaildtiiyta.4ytlRtayitasmfswteedrueerxwipenogisguerahece.hdeaDxnuprdoisonubgrseteahrenverdedcofobovsreecrrlyvienpidecrafiloords,imgoanrltlaslaittey petaontxdihocofiltboyogotynhevtrahelecudoatavieyornfy.oplelAroliwloidrnsag,tealfxlprrooasmustrfheerao1nm.d8tamhtegl/0e.La1te0xtmpwgoi/scuLermgeorworeuerpedwuseraircnergisftaihccerirfeicceodvefroyr paenraitoodsm.iocfAs the asphyxiation and discarded. ed by carbon dicnige MATERIALS AND METHODS A. Test Substance Naasseosf uthae satnucdey;,nHoagevid7e,ncwteoeafsstisnsusubtpaspbtliailneicdtebywywaatshseaossbpssouenmrsveoedrd.taosbepsatlaeblyeeltlhorwousgohlodu.t Pthuerietxywpoasurle a} B. Test Species IaYrnoreiu.vnaRlg,alaediulgth,,mNaolrethCrCalr:oClDina(.SDT)hIeGSratBsRwerartesawpeprreoxriemcaetiveeldy f7orormChwaerelkess oRlidveoLrnabtoheradtaoyroifes, ~ -YY Company Sanfizec. Doonotcontain Techcpr 12E5T77T7:IInnhhaallaattiioonnAApppprrooxxiimmaatteeLLeetthhaallCCoonncceenntraattiioonn((AALLCC))iinnRRaattss__ DuDpupoonn1t212996666. - Rats Crl: have CD( historically SD)IGS BR been used in saf rat was selected ety bas evaluation studies ed on consistently for inhalation toxicity testing. acceptable health status and on Th e extensive experience with the strain at Haskell Laboratory. C. Animal Husbandry I Quarantine `Rwaetrsewweerieghqeuadraanndtionbesdaefrtveerdafrorrivcallinoicar6l sdiagnyssopfridoirsetaostees3titngi.mesD.uring the quarantine period, rats 2. Animal Selection Prreiloerastoedcafcrhomexqpuoarsaunrtei,ne6.maTlhee rsaetlsecwteerdeasneilmeactlesdhfaodrubseeenongatihneisntguwdeyifghrtoamtthaenroartmstahlartatweearned adtetmeomnpsttrwaatsedmandoeovteorrtasnidgonsmolfydgirsoeuapsea,niamnadlsw.ere the appropriate age and body weight. No 3. Identification atEoaccehxhparonasitumrwaeal,stachsoesutiladginlbeoedfaiednaeanctnhiifaimenadilmaafnltuemrabneedxprcoawsghueircecahrawdnadwsedrruereciomnrgadrtehkdeedroenwcioatcvhearrwyadtpeaefrrf-iiioxnde.sdotloutblheepceangse.soPtrhiaotr 4. Housing Eaxbcoevpetcdaugreibnogaredxsp.osure, rats were housed singly in stainless steel, wire-mesh cages suspended 5. Animal Room Environment tReamtpsewreatruerheooufse1d8-i2n6prCox(itamrigteytetdo ttohe22i-nh2a4laCt)ioanncdhaamrbeelartsi.veAhnuimmiadlitryooofms30w-e7r0e%m(atianrtgaeitnededtoat40al6i0g%h)t./daArnkicmycalle.roEoxmcsurwseiroensarotuitfisciidaelolyfitlhleusmeinraatnegde(sfwlueorreeosfceinntsulfifghitc)ieonntamnaganpiptruodxeimaantde/o1r2 hour duration to have adversely affected the validityofthe study. 6. Feed and Water tEaxpcewpattedrurwienrgeeaxvpaoisluarbel,e aPdMIlibNituutmr.ition International, LLC Certified Rodent LabDiet 5002 and 7. Animal Health and Environmental Monitoring Program --~ fAosllsopweciinfgipedroicnetdhuereHsasakreelplerLafboorrmaetdopreyrainodiimcaalllhyeatlotehnasnudreenthvaitrocnomnetnatmailnamnotniltevoerlisnagrperboeglroawm, the those that would be expected to impact the scientific integrityofthe study: TTT eee Company Sanftized. Does not contalnTSCA CBE $2577: InhalatnionaAppproesimate LLetehbalCCoonnceennrmaatbioonn(AALLCC)iinnkRaats DDuuppoonnet1122096666 ~ + Water samples are analyzed `and other contaminants. for total bacterial counts, and the presence ofcoliforms, lead, + Feed samplesareanalyzed for total bacterial, spore, and fungal counts. * bSyamtphleecsagferwoamshferress.hly washed cages andcageracks areanalyzedto ensure adequate sanitation sqCpeuerictiriffeiimeeeddnhatensaivmaynamdefnteoaeltdso,isauefsxleacdte,oexdgiunsa,tracathneltdoermeiadnaxbtiyedmtuhheymdmcraoonncucaferanbctotrnuasrt,eiroanntsdoo fomrekgeaetnysocppeohcnoistfpaihmeaidtnneausnt.rti,tTiihonencalluding `pwroeuslednncoeotbfetheesxepeccotnetdatmoiniantms btpehleaoiwntctehgeritmtayoxfitmhuemsctoundyc.entration stated by the manufacturer alrfahfbeeocratanetidomrtayhleahnveiaalmliadtlihtvayeontfedtriehnneavriisratonu.dnym.eEnvtaalluamtoinointooftrihnegsperdoagtraadmidisnaodtmiinndiisctaetreeadnbyyctohnediattitoennsditnhgat D. Inhalation Exposure System 1 Aunosphere Generation --_ l1C0ah0sasmCbrteoorumnaedtl-mtbootshtpihoetmreseftslsawusebksrptelaangcceeen.deriaFnitletdehrebeaydt,ianhcgirogmshao-nlptirlzeeas.tsiuorTnehoeaftirfh,leamsekstowelraiedsdtheientattsoeutdbhstetofaalnpacperoinxaiimatweiltyh a tt`hhmeeodf0er.ls1t0Sc8mhg5a/1mLbmeearxspwsohsefulrroeew,ac2concothnrdaoemlnlbeserar,tsicowanreiraeeedruostsehodel. evTohlevevdavpaoproprastsherdoufgrhohmeathteedhegaltasessdk ttbuubbyaiinngBg,rionFgfokors hseeactoenddtucbhianmgbienrtfhoer aexnpiomsaulreexpcohsaumrbee.rFwohretrheea1n.w8aaemsrgof/sooLrlmeewxdap.sosfTuorhreem,eatdch.reosvoalpothrepnapsascesdefdrionmtthtehe dTiesscthaatrmgoesipnhteortehsewfeurmeeehxohoadu.sted through an MSA charcoal/HEPA filter cartridge prior to 2. Chamber Construction and Design 3T4h1e.exApobsaufrfelechinasmibdeeresacwhercehacmonbsetrrupcrtoemdootfgedlausnsif(ocrylminddirsitcrailb)utwiiotnhoaftnhoemitnesatlatinmtoesrpnhaelrveo.lume of 3. Exposure Mode `cDvouanrsiicanatgltanecxohpseoedsputiroeect,ehsea.neixmTpaholessurrweesetrcreahiainmnedrbsievrwiedsuroaeltlihynastretershtteerdnaioinsneeodofianpepoaelcryhfmoeartnahityemldamlsetetaxhitanlceernsydslesadtteieenltfoccyelpilnadteerswhwiitchh chamber. the exposure --~ -YY YY `Company Sanftized. Does not contain TSCA CB He25777; Inhalation Approximate Lethal Concentration (ALC) in Rats -- E. Characterization of Chamber Atmosphere DuPont.12966 1. Test Substance Sampling and Analysis aahtpemporsoapxtihmmeoarstpeehwleeyrri3ec0-cdmorniacnweunnttefrraiotntmieortnvhoaelfssHadm-up2rl5ii7nng7g7epawocarhtsetdxrepotoseuurgrmhei.ane2Kd5nbmoywmgnrafvviliotmleerutcmraeicsoafnaclhysaimsbaetr p`Cr-e3-3swMeiicgrhoebdaGlaenlcmean. gTlahses aftibmeorsp(hTeype A/E) filter. The filters were weighsesdetotne tahCatacbonstaaaidned a datimfofseprheenriceesatmhpelperde.- and post-samplriincgcfoinlcteernwteriatgihotnsodfiHv-id2e5d7b7y7t`hweasvoclalucmueloaftedchfarmobmetrhe 2. Particle Size Determination Apaestaimcplleeletsosdtehtaenrm1i,n3e, oparrt1i0cluemsidzieadmiestterri)buwtiaosnt(amkaesnsdumreidnigaenaacehroedxpyonsaumriec wdiitahmeateSriearna1d.peSrocreinnt 210 cyclonepreseparator/Cascade impactor and Sierra series 110 constant flow air sampler.) 3. Environmental Monitoring ohCohmura.omblTelhreeaaainrirdrflfrolewocwowwradasesdsemntoianttiattlohleryeabdengcdionnwtnhiiennnugeaolvfleyrewacichthahneagxepcsoaslwiuebrrreaettemodaaBdcrheioedovukersamtoldeeasltS1903a1irEcahnaanesgebs eper ~ 2hCuhdmiaigmditbiaetlyrtwrtaaecsmepatebarlraegtetuthreeedrwamtaos5m0etta+ergr1e0at%end.dartTe2hco2err+deel2dC3.tiTmhese dtuermipnegractaucrhe ewxapsosimunorgnei.etaocCrhehdae.mxcpbooenstruirrneuealllyawith SBReiHnoScsey1ns0it0aeCtmisdoimngoiwdtaalsptsayrcgherteodmteotbereaantdlreaesctor1d9e%d.at3ivTtehiehmeuosmxidydugirteiynngwcaoensaccemhnetearxsaputoriseoundreww.aitsChmhaeanamsOboemrreeogdxaywgmeoepdneal el 3100R oxygen analyzerandrecorded 3 times during ach exposure F. Anatomic Pathology Evaluation halidlsdtaoinptiaitmhaoolnlsotfghryeoalmnudtnghmsei,0ct.rr1oa0scchmoegpa,/icLnaoleslxeypo(es4xuacrmreionsuesndds.eecrtLwiueonnnstg)a,welgairrgoyhsntsxs,pawaentrhdoeploohbgaiyrayenxsamwienraetipornosaetsseaecdriffioce. sacrificed 1-day after exposure to 0.10 mg/L. ined for (ne 3 quis: ~ -Y `CompanySaniizoc. Doss not contain Tsp car 125777: ntalation Approximate Lethal Concentration (ALCi)nRats ~~ DuPont12966 RESULTS AND DISCUSSION A. (ETxapbolseu1r)e Conditions aeAenriomddaslmsanmwiecretdhieisaxmpsetolusederydsw(teoMrHeM-Ac2o5Dn7)s7i7wdeearrteemde5t.ao5nbaecnodmnac6re.gn8itnuraamlt.liyonrsesopifrab0.l1e0 oinr a1.t8,mgu/tL.eTphseaamtemdosipahneres Smtahlelseerh,igmhorcoenrceesnptirraatbiloenasedruoseool stihzee;choonwdeevnesrat,iiotnaapepreoasSroeelvdefrtoahralmtaittnitgsemlcpaorlugselmdpedrro:e-aa:gcgcl1o0ommpperlroiadstuhecesed, TABLE | : CHARACTERIZATION OF TEST ATMOSPHERES AEROSOLMECAOSNUCRENETDRATION mg/L)" AEROSOL SIZE ~ MEAN 0.10 SD. 0058 MMAD 00R1A8N-G0E15 8] im)' sg PERCENT G2S2D|[2< 4 T | y Spm m [ <iopm 18 012 11-15 8| 43 18 8 37 13 77 86 # ThehfxeepeoisMpuMrreAeDtpsh(oeeMmreanasent,MlosetdaenidnaArfcditdednvyianitagiouinrce.5D.i0aC)m,eataenird)nansbgewfseorroenearc1hopmenxpeaodsperr,iobaisseodon esameplressperexposure, 4 PGeerocmeenttraiecroSstoalndmaarssd.Deviation, la during cach 6Ch8a%mbcehramtbeemrpeariartluroewrraannggeeddffrroomm2135ttoo2245LCi,mcihna,mabnedrtrhelaotxivyegheunmisdoitmyrmanegesd fwraosm2416%to B. Mortality, Clinical Signs, and Body Weights No deaths occurred during the study. ptFoioncnilacoiltwlyiynfogrbeesqxeuperonvsteuldryeiosnb,siwenhrhavelenadttiihonencrlsauttudsdeidweescroweliortrheemdroadtvisesudcnhrarogme otfthehreesntorsanaenrsd tehyeesc.linTiocaelssisgingsnsofare were observed during the remainderofthe recoverydpeerrrieosdtraint. No elimi signsoftoxicity. ~~ - company Sani200.n0otc0o0nt8i car teers ee 125777: Inhalation Approximate Lethal Concentration (ALC) in Rats DuPont12965 "~ No body after the weight 14-day rleocsoevserwyepeerioobds.erved inats exposed 10 010 mg/L immediately afer exposure or tpThhoersreueercoeov,fe6rfryoalptlseroeiwxoedpd.obsyedntoor1m.8almgw/eLigshhtogwaeidn asltieghatpwperiogxhitmalotsesl(y<51-010grgaimdsa)y)ofnorthheedaryomfaotlulgoeweinogc C. A(Tnaabtleosmi2.c3)Pathology Evaluation afTnridsosmeuve3arslauwasetrehedastfurbwomemirtectaescdahcfrriraftoi:mcle3udnag1ts4,dsattyrhasactahwefaet,rrenotshsaeecre(ix4fpicocrseoudosrsen.seedcTtahiyeonasffotlelrow0i.ng10fmigv/eLs eexrpeopsruoreceasnsded ), Laryms, andpharynx TABLE 2 SUMMARY OF MICROSCOPIC FISNDUIBNSGTSAINNCREATS EXPOSED TO 0.10 mg/L TEST ~ D(NAUYMSBAETREERXEAXMPIONSEUDR)E T 14 06 LUNG HNeomtorrermhaargkea,blaelveolar 1 21 Infiltration, mononuclear cell, perivascular - | TRACHEA Not remarkable 33 NOSE Not remarkable 33 LARYNX INnofitltrreamtiaornk,ambolneonuclear cell 213 PHARYNX --NNotoremtarkarbleemarkab3l333e vTehreyremiwneirmealn,oaanpdprweecrieablliekdeliyffaegroennaclescbhaentgweesenoctchuer2rignrgouatpst.heTthiemeaolfveosalcarrihfeicmeorrhages were ~~ aieprlpeehacrooeundsgtiodh0ebreceotdnhceu0rrbersoeunoltfttcohoftnertertoayltpsmeewanentrd.espervoevriitdyede,xptehcetcehdainngethsisprsetsreanint oifnratthse,isasduensoenxeaomfitneedm ------------------ `Company Saniized. Does notcontain Ts ca) H2577: ninAponteLethal Consnenion (ALC)inRas Duron.12966 P Lung weights in 3 rats sacrificed 1 day aftr exposure to H-25777 at 0.10 `mg/L did not appear to be.different study. from control ratsofsimilar `weights in a recent Haskell Laboratory inhalation TABLE 3 FINAL BODY AND LUNG WEIGHTS ONE DAY POSTEXPOSURE ANIMAL BOFDIYNWAELIGHT LAUBNSGOWLEUITGEHT NUMBER 8) 2) 673575 268 1.539 EE 673576 673578 om 262 271 ues 1.540 1.606 = bReolatlivte wLeuingghtWseicgahlt-leFdl BFoodlyloWwseight 100 RELATIVE LUNG WEIGHT (%)* ox 0.57 - 0.59 0.59 CONCLUSIONS ~~ Under the conditionsofthis study, the approximate greater than 1.8 mg/L. According to Haskell lethal concebiration (ALC) for H-25777 is considered to beat most `mass median aerodynam slightly toxic Laboratory toxicity classifications, (aerosol ALC 0.8-2.0 mg/L) in male rats. H-25777 is However, the e6x8pousmur.e.TaHragedttehdeMgeMnAerDiactseddiinaameirenothsaeolrla(tbiMeoneMsnAtusDdm)iaoelslfaerrtehatefttrheoissmtecdo1 ntacoeern4otsBroalmtaittoont,hailsi.tliocsownmceanxtriamtuiomn lwuansg toxicity could have been somewhatgreater. possible that the RECORDS AND SAMPLE STORAGE NSepweacrikm,enDse(liafwaaprpel,icoarblaet)I,rronawModautnzt,aainndRtehceofridnsalMraenpaorgtemwielnltb,eWrieltmaiinnegdtoatn,HaDseklealwlarLaeb.oratory, REFERENCES 1. SCearliceusla2t1i0onAmdebsicernitbeCdaisncSaideerrIampIancsttrourmsenatnsd, CIyncc.l,oBnuelPlreetsienp7ar-a7t9o-r2s1.9IM, Instruction Manual: --- 2. DRautPso.ntUnHpausbkleilslheLadr borat o(20r 03). H) -24678: Two-Week Inhalation ToxicityStudyin Male - Company Sentze. Doesno contain sp a1 eee ----