Part~

Environ mental.Protection Agency40 CFR Part 79SDlsthyl.ne1flamine identification ~Specific chemical Substance and MkU~etesthigRequirements;Final Rule

Dl.thyfert.trlamlne;ProposedTeat Rule;— Rule

ThursdayMay 23~1~S5

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21398 FederalRegisterf Vol. 50. No. 100 1 Thursday,May 23. 1985 / Rules and RegulationsENVIRONMENTAL PROTECTION

AGENCY

40 CERPal799

(OPTS-42012919*4-FRI.2815-Sal

Idantificatlonof SpecificChsnilcalSubstanceandMixtureTestingR.qulrem.ntLD4.thylen.thamln.

AOSNCY EnvironmentalProtectionAgency(EPA)..*CT*Osc Finalrule.

ai~anv:This ruleestablishes testingrequirementsundersection4(a)of theToxicSubstancesControlAct(TSCA)for manufacturersandprocessorsofdiethylenetriamine(DETA~CAS No.111—40-0)consistIng of (1)oralsubchronic (90-day)toxicity in at leastonemammalianspecies,(2) dermalabsorption in the samemammalianspecies usedfor thesubchronic testing.(3) chemicalfateunder aerobicconditions,and(4) mutagenicity(includingtests for bothgenemutationsandchromoaomalaberrations).ThisPhase1 final testruleconstitutesEPA’sfinal decisionconcerningthe testingneedsfor DETA asrecommendedby theInteragencyTesting Committeefor alleffectsexceptcarcinogenicity.

Elsewherein this issueof the FederalRegister. EPA is proposingunder section4(a)of TSCA thatDETA be testedinchroniconcogenicttybioassays,If thissubstanceexhibitspositive resultsin..certain of the mutagenzcftytestsrequired In this final rule.oaiu In accordancewith 40 CFR 23.5(50FR 7V1). this rule shallbepromulgatedfor purposesof judicialreview at100p.m. easterndaylight timeon June6,1985.Thisruleshall becomeeffectiveon july 8.1985.

~esmana~su~noecow~*cYEdwardA: Klein.Directar.TSCAAssistanceOfficeCrS-799).Office ofToxicSubstances.Ra.E-543,401 M St.. rSW.. Washington.D.C 20480.Toll FreeV~(800-424-9065).In Washington.D.C.(554-1404).Outsidethe USA:(Operator.202-554—1404).su~.suvrrasvNWOSMAnOseIn theFederalRegisterofApril 29,1982(47FR18386),EPA issueda proposedruleunder section4(a)of TSCAto requiretesting of DETA for avarietyof healtheffectsandfor chemicalfateunderbothaerobic andanaerobicconditions.Today, undersection4(a)of TSCA. EPAIs promulgatingafinal PhaseI test rulerequiring health effectstestingand

chemicalfate testing(underaerobicconditionsonly) for DETA.1. Introduction

This notice ispartof the overallimplementation of section4of the ToxicSubstancesControlAct (TSCA, Pub.L.94.-469,90 Stat2003 etseq..15US.C.2801etseq.)whichcontainsauthorityfor EPA to require developmentof data

For a more completeunderstandingofthestatutorysection4 findings,thereaderis directedto theAgency’sfirstproposedtestrulepackage(chloroaietb.neandchlorinatedbeuzenes.publishedJuly18.1960(45FR48510)1and to the secondpackage[dishloroinethane,nitrobenzene,andi.1,1trfchloroethane.publishedJima 5,1981 (48FR30300)) for in-depthdiscussionsof the general issuesapplicableto this action.IL 3ad~powid

A.Pt’Ofile

relevant to assessingthe risks to healthandthe environmentposedby exposureto particularchemicalsubstancesormixtures.

Under section4(a)(1) of TSCA. EPAmustrequiretestingof achemicalsubstanceto develophealth orenvironmentaldata if the Administratorfinds that:

above.DETA reactswithCO, in the aito form carbamateswhich precipitate‘from solution. For this reason,DETA i~manufacturedIn anessentiallyclosedsystemand is transported underanitrogen atmosphere.Due to DETA’sreactivitywith airandto the conditiorusedduring its processing,the Agencyconcludesthat it is unlikely thatsignificantemissionsof DETA to theatmospherewill occur. On the otherhand.EPAbelievesthat significantreleasesof DETA to waterwill occurduring manufacturingandprocessingoperations.EPAbelievesthatoccupational exposureto DETA(primarilyby thedermalroute) occursduring manufacturingstorage.transpprocessing,andclean-upactivities.a,

• that themost likely sourceof consumexposuretoDETA consistsof dennalcontactwith epoxy-resinproductscontainingthe substanceasacuringagent.L JTCRecommendations

(A) (i) th. manufacture,distribution in commerce,proc.eonng,use,or disposalof achemicalsubstanceor mixture,or thatany combinationof suchactivities,may presentanunreasonableriakof injury to healthor theenvironment,

(n) thereare insu~cientdataand.aqerienceupon which theeffectsof suchmanufacture,distributionin commerce,processing,ma,or disposal.of such substanceor iI~iThweor of any combine-tion of suchactivitieson healthor thf.Invironmentcan reason-ably bedeterminedor predicted,and

(di) testingof suchsubstanceor mixturewith respectto suchsSsctsisnecessaryto developsuchdata;or

(B)(i) a chemicalsubstanceor mixture is or will be producedinsubstantialquantities,and(I) it eatenor may reasonablybeanticipatedto entertheenvironmentin substantialquantitiesor(II) therein or maybesignificant or substantialhumanexposureScsuchsubstanceor mixture,

(ii) thereare insufficient data and experienceuponwhich theeffectsof the manufacture,distribution.incommerce,processing,use,or dispostiof such substafleeor mixture or of any eombina-tion of suchactivitieson healthor the environmentcanreason-ablybedeterminedorpredicted,and

(iii) testingof suchsubstanesormixture with respectto sucheffectsis necessaryto developsuchdata.C

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DETA. CASNo 111-40-n.Is s~alkaline.hygroscopic~viscousliquid.TheestimatedannualproductionofDETA in 1982rangedfrom 28 to3~.million pounds.The primaryusesofDETA are for theproduction of paperwet-strength resins,epoxy-curingagents.cheIati~gagents,lubricatingoilandfueladditives,surfactants.andcorrosionInhibitors. DETA alsohataminoruseasa decontaminantformilitary chemicalagents.As discussed TheInteragencyTestingCommittein detail in the Agency’s - (lTd. organized undersection4(e)o~Diethylenetriamine Support Documeni. . theToxic SubstancesControl Actwhich is available from the TSCA tTSCA), Included DETA in its EighthAssistanceOffice at the addressgiven . Reportto-theAdministratorof EPA

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FedaaaLRegister/ VoL 50, No. 100 1 Tht~rsday,May 23. 1985 / Ruins and Regulations 21~

April 2& i9~1.p~SdE’thSF.dmeLRegisterof May 23.1961(46FR23139).The ITC designatedDETAa.aprioritychemicalandre rw,i.~sndadthatit betestedfor healtheffects,tochroniceffects.repro~0I&~-E~t~. andteratogenLciLy~TheLTd basedittdesi~*tionof13E~Aon thesubstancesknownbioiogical efTacts,thereported

• produclianinexamsof10milhiocpoundsperyear.andtheNationalOccupationalHazardSurvey(Ref.1~estimatesthat63,000workersare.potentiallyexpoosdtcDETA.

CPru!~• EPA IssuedaproposedrulepubLhedIn the Fss~afXApr~2L1I6Z(47FR18396)in ~ lathe instfr~recammendatlcrssbytheTICon~rA.

1. TestR U1c~JaE~The pru,u..edrule requiresthat DE’TA betastedfor~

a.Subchroaic(g0day} healtheffectsIn at least two man~han.p...

b.Mutagen~cltyf~enematationandcy ___

andanaerobicmaditiessj.TheEPA basedIts noposedtesting

reqm~me!~. ondieauthorityof section4{a)(1)(AJofISCA.

2. Findingn TheAgeecyfosodthatthemanufec~e.~ ass.anddisposalofDETA may presentanunreasonablerisk of injury tohealth,dnsin ~ andiwsagenfaeffects.for thefollowing resson~

a. There ma e~istlng~te winchIndicateapoinutial~n healthhazardh~DETA Mth re.~.atin theseeffects.

b. EPAbelieves~ eraexposedin DfFA In the kpl~.inusingcenaiwsaproducts.~of releaseoi DETA intoenviron~-

C. The~ecp alsofe ~t ~are~enr~t datain l~2 thesubchrsoie d~qc clTi~~fDETA. ~ inedugoL~Ain~-----—,~ d_~

in andi~a.~A ~ the~-- —i. ass.

disposalof DETA p—&unrea~l* i~ to ~ ~bktoon~ ~4Lc~ s&L~ta~by theN-nlfrouminederivative of DETA. for thefollowing

d.ManyN.eitios~~hewbeenshownto besereinogenlc.1~-ar.existing~e whishindicatoatheoreticalpotentialfor theconviwianof DETA in .N.ait,osmaienin theenvfronmailandthet~ona mapbeexposedtothi.AT-ultr.samineasareseMof thesuleas.of DETAindi.envfrenmenL

nTh.date..,~ien&to predictthe~ of

resultingfrom DETAreleaseto theenvirr~nt. and ch~i.&lLate testing(under both aerobicand anaerobicconditions)isrV’.c4~ tcdevelopsuchdata.

3. Differencesfrom 17’C.gRecommandaticna.lithe proposedrulefor DZTA.theEPA alsoprc~~thereasonswhy theAgsocyspro~e.edtesting.ieq~akemeninfor DETA differedfrom the TIC sosmameedabenahethe

_______ U follcwsso.L~Acyproposaksuhehreinc

testingridesthan -iletini, ~eeicstodion became theAgeeqlmMe,sathes~p—~y ~ 60.diy~withse~~mi%mmayb.ssodan...________Iifeti~estoc~at~aMeffectsother th~~riri~p ~certainother age-relatedeffects.Theavailable data providedno soundbansfor supensise.CDETAs’abilty to case.oncs~~orags-islatedeffects.

b. EPA~Enot prepmetestirqfarreprsdactiwsad~ effects.becasae,hr ~Agmay”, ~ di.availabledata çaMIr.~limited)didnotsuggest.poinntia~hatheseefisota.

The analysisand find~onwhichtheabovedeterminationswerebasedirepreen~trdfri the~eby1anetit~neSupportDo.~nt.whichIs aenthblefrom the OfIn,olTcxfcSithstancus’‘rSCA A~efatancaOffionTherrc,recommendati~andEPA’S~testingrequirementsmabelow~

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4.bsassfrrC~~aizrPr~,po..dRule,In th. proposedt.sti~ for D~TA..the~ecy raisedthefoliewingm.~ceIssuesfokc~~w.~i~

eShedintydeonodiarethyIensia~~~atethyIenedi~esodfri.thylenstetsamine}beusedseasurrogatefaaDETAtonicity?

b. Whetprotosni.s~eI~beusedfezthe chemical faintes~ofD~E&~n.d&t quantifyingdeextentof debiological (or chemical)lyaaslormaklgnof DZTA to anIJ.introsaminederivativeofDETA bysir zgsaisxanpres~Inwater. sew~.andsail?

c. Becamaof dithouMiasInvolvedInquantitmingthedeseQf~A thetanimals oofd~ve.in ~aAsubchreele~96.day).t~,, ~f~gI’~is-

not the Agency’schoiceof thema!routeof exposurefor therequsud scbchroni~studiesappropriate?

d. Aithounji theAgency is spectfyiagthe oral route of exposurefor- therequi,ediubchronic[90.day)testingofDETA. theAgency is primarilyconcernedabout dermal expos~’.vtothis chemical:would it. therefore.not benecessaryto requiretheperformanceciadermalab.rpttonstmiyof DETA toprovidedateaesdedin e’uabeststherisks posedby dermal expoasond

s.A1i1 theTIC ie~~~-~’hill-~eti.~ hsla8ofIJETA.would nil ~k1~nd~(96-day)tanicitytestingin”-~g~ cespr~nAvehistopathokgisel———-—ticsofbodytissues,be.d~- b~.ce~LETA-related effectsthatwouldbeob.......,IIn fu1J-hfe~e~ 1rut1x~.~ hethosee~.sinre.,Jii..~long latencyperkida?D.NewDev’~’~ Fo&ProposedRule

Theproposedrule far DETA ~‘PR1838&Ap,Rm7961~indicotedth.t.lfinterestedpertlea,~uoded—•opporr~eyfaroral ca~e~codieproposedrefe. then theAgeniy~M

ameetingenthisr~ toWaahIngtee~DC..enJuly 13.1962.Sinceno requestswere receivedby the EPAfor thepresentationoforal commentsonthis rule, anpeb cmeetfugweaheftAmeetingbetw~en,.-..ntatlvesfromthe Union Carbide Corporationendmembersofth sdenti~cstaffof theOffice of Toxic SubstancesofEPAheldanSeptsmberlO198Zin dIscus,mutagenicitystudiesofDETA (1ef 3)performedfor that armiofactusenwhichare discussedin datailIn Unit 1IL F.

1.kidss~yComments.The DowChemicalCompanyaridtheUnionCarbideCorporationsubmittedthefollowing additionalrelevants~n intheAgencytogetherwith theseSrmVcomments,datedJune ~ on theproposedtestrule for DETA

a. Structure!and~ologlcalActivityRelationshipsBetweenEthylenedlamineandDiethyfenetriamine(includedpreliminaryabsorption.distribution.metabolism.andpharmacokineticsstudiesof ‘4C-radiolabelled DETA andethylenediaminefollowing oral orendotrachealadmini.tratfonto malerata}.

b. A~dascrIpthmofstudiesperfomed.byDow Ch,,nit’i~1 Companytodetermineifs Nm’trma~ederivativeof DETA would form in anequeaorinvitro chernicaL~de1system.

2.SectionS(d~lS~efucm2~29&3. In. a.TSCA “ ~ ~on DETA rscaLvadbydeAgan~on.

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21400 FederalRegister/ VoL 50, No; 100 /. Thursdtiyv May 23, 1985 / Rules and Regulations

June24.1963,UnIonCarbideCorporation included the followingadditional relevantreportm

a. PharmacoklnetlcsandMetabolismof Diethylenetriaminein theRat.(Acompletereportof preliminarystudiesInItem l.a.).

b. Dlethylenetriamlne-ComnrerclalDermalCarcinogenesisStudyIn MaleC3HIHeJMice.

C. Diethylenetrmamine-HIghPurlt~DermalCarclnogenesisStudy in.MeIbC3H/HeJMmon -

3.Aai5ctiontd)SubinisslonzMarch ~ bra~submissionusDETA receivedby~.AgencyenMarch22.19S4.UnionCarbideCorporationIncludedthefollowing reportandassociateddocument

a.Summaryof ExploratoryTestsatBRRC (Bushy Run ResearchCents,)with the CHO/HGPRT (Chinesehamsterovary cells/hypoxanthine-guaninephosphoribosyltransferaselocus)TestSystem.

b.A memorandumto Mr. Di..Heywood..UnionCarbideCorporation.from Dr. Ronald S. SleuinskL BushyRunResearchCenter. desathiaghow theresultsof the abovestudy relate tovarious EPA guidelinesand U.S. EPAGene-ToxProgram reports onthis assaysystem.

Items l.a.and2.a.have beenjudgedby theAgencyasinsufficientevidencethat toxicity data for ethylenedlaminecan substitutefor the requiredtoxicitytesting of DETA (seediscussionin UnitlILA.).

4. EPA ReviewofSubmittedBioassayData. The Agency has carefullyreviewed the two dermalcarcinogeulcitybioauayaof DETA(DETA-CommerclalandDETA-HighPurity) submittedby the Union CarbideCorporation(Items 2.b, and2.c.), todetermineIf thesestudie.wereperformed in a mannerwhichwouldnegatethe needfor oralsubchronlc(90.day)testingof DETA. TheAgencyhasconcludedthat thesestudies.areinadequateto negatethe need for oralsubchronictasting for thefollowing /reaions

a.In bothof thesubmittedstudies,only male micewereused; thus, sex.relateddifference, In responseto DETAadministration couldnot beinvestigated.

•b~In bothstudies,onlyasingledismaldosagelevelof DETA wasemployed;thus, no dose-responserelationshipswith respectto DETA.relatedeffectscould be Investigated.

c. TheAgency ii requiringcomprehensivehlstopathologlcalexaminationof tissuesin the required

- oral .ubchronic(90.dayjtestingof

DErA.~bothof the submittedstudiescontainonly very limited histologicaldata.

TheAgencybarreviewed the dataonDETA contained in Items3.a. and3.b.,andha,usedthesedata in reachingconclusionsregardingtheability ofDETA to Induce specificlocusmutations(atthehypoxanthine-guanin.phosphoribosyl-transfereselocus)inChinesehamsterovaryusia(seediscussioninUnitIILF.). In addition,theAgencyhasreviewedtheinformationonDETA containedin lion tb., andhas

- concludedthatthesedatedonotnegatethenicesnityforch.miea-Ifats tastingciDETA todetermineif saNos~~fr~derivatlv.of thissubstancecouldbeformed underenvironmentalconditions(seediscussionIn Unit WI.).ilL PublicComm~t’

Thecommentsreceivedby theAgency in responseto theproposedtestrule for DETA werefrom theaffectedIndusty and tradeassociationsources.The majorissuesidentified duringthecommentperiodarediscussedbelow.A. AppropriatenessofUsfr,gAnalogueDatato AssessDETA.’, Toxicity

Oneof themajor issuesfor which theAgencyrequestedpubliccommentin theproposedtestrulefor DETA Is theappropriatenessof usingavailabletoxicological InformationonethylenedIa~~~in~(WA) andthethylenetetramine(1TrA). proposedstructuralanaloguesof DETA... asubstitutefor testdataon DETA itself toassessthepotentialtoxicologicalhazardsposedby DETA. The two majormanufacturersof DETA in theUnitedStates,theDow ChemicalCompany(Dow)and theUnion CarbideCorporation(Union Carbide), submittedth. onlycommentsaddressingthis Issue.ThesemanufacturersbelievethatWAI. a closes~ucturalanalogueof DETA.statingthatDETA may beregardedasthedirnerof EDA. andthat bothsubstancescontaintwoprimaryaminogroups,while DETA (but not WA) alsocontainsa secondaryaminogroup.DowandUnionCarbidebelievethatthechemicalbehaviorof both DETA andEDA will be dominatedby thepresenceof the two terminalprimaryaminogroupsIn thesubstances.Furthermore.thesefirms pointout that the physicalpropertieswhichInfluencebehaviorInbiological systemsarealsosimilar forthe two substance.:both irecompletelysolubleInweter.bothformbasicaqueoussolutions,bothayerelativelynonvolatile, andbotharelow withrespectto molecula,weightThesemanufadw’~ribelievethat,becauseofthesesimilarchemicalandphysical

properties,thesetwo substancesarelikely to behandledsimilarly In -biological systems.

DowandUnion Carbidesubmittedtiresultsobtained from studiesusing ‘~CradiolabelledDETA and WA andaimedat determiningthe excretionpattern,the tissuedistribution, andtheblood-levelpharmacokineticsof theradioactivityobservedfollowing theoralor sodotrachealadministrationofthesersdlolabellèdcompoundstoFischer344 rats.In addition,theurinarradioactivityobtainedfollowing theedmI,~L~trationof rsdlolabelledDETAandWA toratswascharacterizedby1on.evrl~engechromatographicmethniDowandUnionCarbide interprettheresultsofthesestudiesasIndicatingtiDETA and WA have the urns generapattern of disposition In rats. andthatDETA andits metabolitesaremorerapidly eliminatedfromratsthanEllA

• andIts inetabolitu.Thes.firmssubmittedto theAgencythe resultsofbotha7-dayrange-findingfeedingitsin rats of the dthydrochloridsofWA..and the results of a 3-monthsubacutefeedingstudyof WA dihydrochhatdsrats,andbeliev,thatanextensive.toxicity testingprogramfor DETAshouldbedelayeduntil th. resultsoffurther toxicological testing.onWA.becomeavailable.

TheAgencymustdisagreewith themajormanufacturersof DETA that tb~proposedtoxicity testingof. thinsubstanceshouldawait furtherInformationregardingthe toxic effectof EDA. While DETA andWA aresimilarin thattheybothpossesstwoprimaryaminogroups.they arevastlidifferentwith respecttothe factthatDETA alsopossessesasecondaryaminogroup, while WA doesant.,Substanceshavingsecondaryamine.groupsarewell knownto bemuchansusceptibleto stableN.nitrosamlneformation than substancespossesath~only primaryaminogroups.Other.uyet unknown, differenceswith respssthe production of toxicinetabolitesnalsoexistfor substancescontainingsecondaryaminogroupsasopposedthosecontainingonly primary ammogroups.

In addition, studiessubmittedbythesemanufacturersdo not,In fact.demonstratethat themetabohtas

• producedby ratsfrom DETA and El~arethe same,sincetheradioactivemetabolitesappearingIn the urine

• following the administrationof thetiradlolabelledsubstanceshav, beencomparedonly by ion-exchangecoltchromatographictechniqueswhich,1

-. themselves,do notallowstructure!Identifications.Even thiscomparisot

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FederalRegisterI Vol. 50, No. 100 / Thursday,May 23, 1985 I Rules and Regulations 21401

flawed by the fact that different elutionsystemswereemployedin thecohminchromatographyof the radioactivityfoundin theurineof ratstreatedwithradiolabelled DETA or WA, makingcomparisonsof radioactivemetabolitesby their columnelutlonvolumesimpossible.In addltion..thecolumnchromatographicprofilefor theradioactivityfoundin theurineof rat.treatedwith radiolabelledDETAcontainsat leastsevenradioactfve~peàfractions, while the.correepoodi~chromatographicprofile’ for the--~.radioactivityfound in the urine ofrats -•

• treatedwtth radiolabelled~A r~mtaiiig• only threeradioactivepeakfractions. -

This fact maywell indicatethatDETAis, in fact. metebolizeddifferently thanWA by: therat,In anycase,thedatafrom thesestudiesdonot Indicate thatthemetabolitesproducedfromDETAfollowing oral or endotrachealadministratiotito Fischer344 rats.siethe sameas thoseproduced under thesameconditionsfrom FDA~.Evenifdatawere available to demonstratethat themetabolitesderwedfromWA wereidentical to thosederivedfrom DETA.theAgency could not,accept •, , -

• toxicologicaldata on WA asa . -- substitutefor thetestingof DETA ~tielf.

becausethereIscurrentlynoacceptedmethodologyfor determining thepotenaesexpectedfor DE’tA for thehealth effectsof concern from analogue

• data availablefor WA or otherpotential analogues,

On the basisof radioactivity alone(sayingnothingabout thechemicalstructures actually containingtheradioactivity), themetabolism studiesinthe ratsubmitted by Union CarbideandDow indicatethat bothDETA andWAarereadily absorbedvia the oral andendotrachea!routes,distributedthroughout thebody,andexoretedprimarily in theurine andfeces.Thesedata also indicatethat DETA anditsaretabolitesareeliminatedfrom thebodyata fasterratethanED&andIts

• metabolites,and.In addition,thatDETAand/oritsthetabolltà~ar.retainedto a

• lesserdegreethan-WAmd/OrItsmetabolitesIn bodyI4.~.& •

In sathe-marderlyingmechaniiin, responsiblefor thetoxiceffectsnoted by Pujino{ReL.2)..primarily In the livers,,kidneys,andlungsof Wistarratsreceivingchronic

- treatmentwith DETA via the dermal or-‘ subcutaneousmutes.areasyet

unknown. Tho Agencyhas noevidence• to indicatethatthe metabolites

producedfrom DETA by.rste(whichmay be.responsiblefor someor all of theobservedtoxic effects’of this substance)are the samemetabolitesas those

produced from WA by rats. In addition.even if data were available todemonstratethat identical metabolite.were produced from both DE7A andWA by rat., analoguedata availablefor WA cannot currentlybe used toarrive at expectedpotenciesof DEFAfor thehealth effectsof concern.Therefore, basedon the aboveconsiderations,the Agency hasconcludedthatit would beInap~~pil.teto utilizethetoxicity dataavailable for WA (or otherproposedstructural analogue,of DETA. suchasTETAJto assessthepotentialtoxicologicaihazardsposedby.DET& -andthattestingof.DETA Itself Is.necessar~. -B.Appropriatenessof SabchrvnicRatherthanChronicTestingofDETA

Mother Issuefor whichtheAgencyrequestedcommentsin the proposedtestrulefor DETAin theappropriatenessof a 90-daysubchronlc -testdurationratherthanafull-lifetimechronictest durationfor theproposedtoxicity testingof DETA. Thejnsjoymanufacturers-of DETA In theUnited.States,DowandUnion Carbide.submitted the only commentsonthisissue.Theposition ofthesemanufacturersIs thatanoral90-dàysubchronicstudyof DETA will sufficetoprovide adequate informationonallsystemictoxic effectswhichwould beobservedfor thesubstanceIn anoralfull-lifetime chronicstudyof thissubstance,with theexceptionofcarcinogeniceffects.The Agencyalsobelievesthat, In general, a properlyconducted90-daystudy,Includingcomprehensivehistopathology.can beusedasa surrogate for the full-lifetimechronicstudy with respectto thedetectionof chemical-relatedhealtheffects,exceptfor thoserequiringlonglatencyperiods, suchas carcmnogemuity.Therefore, theAgencyIs requiringsubchronic(90-day)testingin thefinalPhaseI testrule for DETA. In lieu of full.lifetime chronictesting. -CA~bpropriotenessoftheOralRouteforSubchronicTestingofDETA

In the proposedtestrule for DETA.theAgencyalsorequestedcommentsonEPA’s selectionof the oralrouteofexposureasthe routeof choicefor therequired90-daysubchronictoxicitytestingof DETA. Althoughthe Agencybelievesthatexposuresto DETA willoccurprimarilyby thedermalroute,the

-difficultiesassocm,tedwith determiningtheactualdosesof the testsubstancereceivedby theanimals In studiesutilizing this routeof administration.togetherwith the factthatpreliminarypharniacoklnetlcsdatasubmittedto

EPA by Union Carbide indicate that,DETA is absorbedfollowing oraladministration, led the Agencytoconclude that theoral routeofadministration should be required forthe subchronic testing ofDE’FA. OnlyDow andUnion Carbide commentedonthis issue.Thesemanufacturersagreedwith the Agencythat oralstudiesofDETA would allow the adequateevaluationof thesystemictoxicity ofthis substancewithoa~thedifficulties ofdetermining the effectivedosesreceivedby treatedanimals which would arise Indermalstudies.In addition,thesemanufacturerspointedoutthatthe-knownskin Irrttancy andsensitizationpotentialsof DETA would likely leadtostressfulconditionsIn pniTn.l$ receivingDETA by thedermalroute,making theevaluation ofthesystemictoxicityobservedin suchstudiesdiffiCult Thesedifficultieswould not aria.In oralfeedingstudies.Therefore,theAgencyI.requiringoral90-daysubchronicsoximty -testingIn thefinal PhaseI testruleforDETA. •

D. NecessityofaDermalAbm,pcà,nStudyofDATA

AnotherIssue.whichtheAgency -raisedfor commentin the proposedtestrule.forDETA wasthepossiblenecessityof requiringadermalabsorptionstudy of DETA. sincetheAgencyis primarily concernedaboutpotentialhazardsposedby thissubstancedue to exposuresvi. thedermal mute.Only Dow andUnionCarbide commentedon this Issue,andthesemanufacturer,believethat suchadermal absorption study would, Indeed,benecessary.in addition. thesemanufacturerssubmitteda protocoltotheAgency for astudyaimedatdetermining thedegreeof dermalabsorption of DETA In rats from data-obtained in dispositionstudiesof thesubstance,using both Intravenousanddermal routesof exposure,

Sincehumansareexpectedto beexposedto DETA primarilyby thedernial route, theAgencyconcludesthata dermal absorptionstudy of DETA Is.In fact, necessaryin order to assessthehazard, posedby DETA by this routeofexposure,andIs, therefore,requiringsuchtestingin thefinal PhaseI testrulefor DETA in the samernamm~flanspeciesselectedfor the requiredoralsubchronic(90-day)testing..£ Protocolsfor RequiredChemicalFatsStudies of DETA -

Thefinal Issuefor which-theAgencyprquestedcommentsIn its proposedtestrule for DETA involvedwhich protecolsshouldbeusedforthe aheetica}fate.

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studiestheAgency is requiringforDETA. SincethemiciobielformationofN-nitrosarnitmesfromsecondar~aminespresent in water.sewage,.andsoAlhubeenreportedby severallavestigeters(Ref..5 thruugh71~.tbmAgencyisconcernedthatanN.ni~o.a1.1l1t.derivative of DETA aa~be firmed Intheseeiwiiooments~upeciallyinsewagetreatmentfacilities),may betransported,to waterenircesusedfor -the pco~4a~t4nnof king.w,ten..~,suavivedrinhing-water treatmentprocsds.aud.ihus.~a potential-carcinogemchawdto thegeneral -publicby its presenean’4’~n1’ngwatonA clom sthic*maral enelegeeof theN-nhtrosaminederivathsdDEl’A,N.fliodith~n~~Lsaw3eL1~ELA),is akno~wt~ cerc1msen(Rd..athrough-ID). In sdcbboe,Yerdy andAlexande~Rd~.Jhaea4—~tr.iedthatNE~LAwa.foxmedfromdletKanoIaminein lakywatarend-Insewage,andthataileaaLsoasoIthis-ND~.Aprcelucboawasprobablydue tothe actionof micmoorganismsor someotherheat-labilefactor.ThesedataalsoIndicate that someofthe ND~.Aproductionwasdueto purelychemical(nonbioIo~cal)react~WhethertheN-uAliosamin.derivativesofdiethanol.~vth~~ci DETAare formed bypurelychemicalorbiological meanshas.no bearingon thetonichazardspøssd.by theN.aitro.aminederivativesthemselves.Therefore,theAgencyproposedchernicalfat.testhngofDETA.underbothaerobic andanaerobicconditions,to quantifytheamountofthepotentiallycarcinogenicN-oafrosaziinederivativeof DETA whichmight formfrom DETAIn water, wage.andsoil.TheAgencyinvited commentsonexperimentalprotocolswhichm~tbeusedfor this purpose.andreferredinterestedpartiesto themetbodol~esreportedin thestadiesof Yordy andAlexander(RL 7).

CommentsWerereceivedon thisleg.,-from Dow,UnionCarbide,endtheChemicalManuftcterer~-Ass.datfen(Q4A).Dow andU~ Carbidebehavethat theproposed~ tasting.of DETA aimedatds~~g if aN.nifrosaminewill beproduced~DETA dueto aerobic oranaerobicbiode~adadonIs eossuary.With It.comments,Dowsubmittedadesaiptionof anunpublishedpreliminaryin vitrochemicalstudy fu.~ by Dowaimed.atdet~~rminlngif anN.nitrosamineof DETA would formInaqueoussolutionwhenNO1vaporwasbubbledthrougha50percentsolutionofDETAin D,O (usedfor-nuclearinagnetiC-resonancepurposes)underaerobiccondli.ms.Uain~VItMYhOIM sailnuclear

juagneticresonancespectrocietry.Dow- concluded from this preliminaryexperimentthatalmostall of theoriginalDETA haddisappearedfrom thereactionsolution,butthatno-characteristicUV absorptionbandat300-400nrncouldbedetectedtoindicatethepresenc.of anifrosainlne.Dowconcludedthatit is unlikely thattheN-nitros~reof DETA mm beproducedunderaerobicconditicos.or. Ifpmduced.theNoifrosamnlneaI~lAmustdomanpomrapidly.

With respectto theanaerobicbiotransformation ofDETA to anN-njtrmmminederivative,UnionCarbide -andDowb.lievethistobshlgbly -unlikely, sinceDETA (which isverywatersoluble)wouldnotbeexpectedto-~sorb to soilsandserlimAnil having...anaerobicenvironments;additionally.thesefirms believe(citingRefs.12through13) that anynitrites presentInsuchenvironments wouldbemetabolizedby mioroorgamsinsto’nitrogengs~thusremovingarequiredreactantfor nitrosamine formation.

UnionCarbide andDow believrthatchemicalfate testingfor the fôm ation ofanN-nifrosamlnederivativ, of DEMbymi~oorganismsunderaerobicconditionsis afar reachingresearcheffort whichIs notwithin thescopeof.,TSCAsectIon4 testride.Thea,firmsclaimthatno standardmethodologyfordetermining low concentrationsofN-nitrosamlnesin water currentlyexist..andthat the synthesisandcharacterizationof the N-nifrosaihlnederivative ofDETA would be required.prior to the developmentof ananalyticalmethodfor thedetectionoftheN-altzosaminaderivativ, of DETA Inwater at thai ugh leveLIn addition,Dow andUmanCarbide believethat th.work of Yordy andAlexander(ReL 7)1.Insufficientasa modelto usefor thebiotransformation study.sin~ethemethoddoesnotdistinguishbetweenbiological andchemicalproduction ofN-nltrosamines,andthatanew. -methodologywould haveto bedevelopedfor studyingthe -biotransformationof DETA to anN-

“nhtrosaminederivativeandarrealenvironmental,ww4l*~~.

UnionCarbideendDowsee.nnnertainas to whetherEPAIsproposingtorequirequalitativeorquantItativechemicalfsta.testlngof DETA ~ -det’i.ninb~gIf theN-rduosamlnaderivativeo(DETA would befcemadbymlaoorge”{~a.underaerobic andanaerobicconditions.Thefirma believethat, with respectto aerobiccondiioce,theprutIimin~~yin vitro ~n1~aI ttlngby Dow~ur ~rAMsufficientto statsthat either.en14-

nitrosaminederivativeof DETA doesnot formor.if formed, rapidly degrades.Thesefirms also believethat thescopeof work involved in answeringthequestion in aquantitative fashion is toogreatto bewithin theproperscopeof aTSCA section4 testrule.

With respectto this issue.CMAcommentsthatEPAshouldavoidrequiringtestingunderTSCAsection4in caseslnwhich thetestingmethodelo~fsnot sufficientlywelldeveloped.Specifically,OdAbelievesthat thechemicalfatetestingrequiredInthe proposedrule(aimedatdetermininglienN-aitmuminelsproducedhoesDETA by’mlcverganismaimdsraerobicorarmarobicoondltions)1. -.Inappropriate,simc,to DlL’~s - - -knowle~,thereI. ao~rentstenderdmethodologyfoi’pcf.~.gthesetests.CMA belleveehtks*theresultsfromthesetestsweunlikelytobesufficientlyreliablefor u.s-inEPA’sdedEoelongrthv~G~rw.suth~tes~fellsoutsidetheanop.of ‘tvhat canbe-requlrod-under-TSCA.

Certzlnof thecommentsde.mibed-’aboveIndicate-uncertaintyor~~diieiOzr’concerningtheAgency’.rat1onali-~theproposedchemicalfate testWatDETA. asde,albedin thepropesedtestrule~ titlesubstance.In-actuality,theAgencyis concerned-aboutthetotal’transformation.whetherb*ologfcal’or’-piselychemicalin esters,ofDETApresentIn water,sewage.or’soils teenN-nltrosaminederivativeof-thesubstance,whichtheAgencyviewsas-apotentialcardumogensadwhichmay - -.enter thedrinldngwater~

With this clarificationregardingtheAgeucy~sconcernsaboutthechemical-fatetestingof DETA, EPA disagree. -with bothDow andUnion CarbIde~aswell as CMA, thatappropribte -methodologydoe.notexist,orbe easilymodified. for the successfulcompletionof this testingrm~uii~ent -In addition, theAgencydisagreeswiththemanufacturer,that the-r.sultsof thechemicalstudiesperfonue~Iby Dow.aimedat determiningIf anN- -nitrosaminederivative ofDETA could-be formed fromDETA in aqueoussolution,obviatethe need for’ theproposedchemicalfate testingof DEM.DowconrInd~i4hoertheses~esthatIt is unlikely thatanN-u.lfrosaminederivative of DETA wilt form In aqu~ssolutions.or If It doesform, It willdecomposerapidly; Basedonthebriefdescriptionof thesestudieswhichwassubmittedto theAgency(no protocolforthesestudiesor final study reports weresubmittedby thenwaufncturan),EPA -believesthatanN.rdfroninsderivative aiDETA did, Infant fa~bu1

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decomposedwider the exper.mentalconditionsemployed.The fact thata N.nitrosanune derivative of DETA can,indeed,form In aqueoussolutionsIsdemonstratedby thestudiesof Popp(Ref.14), In whichDETA wasdetectedin waterby chemicaltraceformation ofhis substanceto aN.mfrosaminederivative, which was.subsequentlydetectedby-polarography. The fact that.theN-nitrosaminederivative of DETAprovedto beunstablein Dow’sin~Pitivchemicalsystemdoesnot Indicatethatit wouldnecessarilybeunstableunderenvironmentalconditions.Yordy andAlexander(Refs.7 and15)haveshownthat asubstanceverysimilar In -chemicalstructureto theN-nlfrosammnederivative of DETA. N-nltrosodlethanolanulne,wasessentiallystableto degradationIn someenvironmentalwaters,especiallyduringthe wintermonths,andwasslowlydegradedInothers, primarily duetomia’obial metabolism.In addition. TateandAlexander(ReL 16)demonstratedthat threealiphutic N-nitrosamines (N-nitro.odlmethylamine,N’nitrosodiethylamlne.andN-nitr-osodi~n-

- propylamine).whichmaybe viewedasanaloguesof the N-nitrosamlnederivative ofDETA, wereresistanttodegradation In soil, sewage,andlakewater. No degradationof theseN-nitrosamineswasobservedin lakewaterduringa3.5-monthperiod. Thus.’theresults of Dow’sin vitro chemicalstudy do not, In fact, obviate the needfor the chemicalfat, testingwhich theAgency is requiring in the final PhaseItestrulefor DETA.

The Agencydisagreeswith themanufacturers’contentionthatnomethodologyIs currentlyavailablefordetermininglow concentrationsof an N.nitrosarninederivative of DETA inwater, andrefers thesefirms to thepolarographicmethodof Popp (ReL 14),thedetailsof whicham unpublished.butwhichIs based(EeL‘17) onthepublisheditudlesof thepolarographicdetectionofvariousN-nilresamlnesbyDahmeruatct(RILIO) andOmangandHagtca~(Ref~.10),Although Popp(ReL 14)haddemonstratedthatanN’nitrosamine of DETA can, In-fact, be -~detectedby polarographlctechniques,the AgencyIs aware thatcontaminantspresentin the environmentalsamplestobeutilizedIn thechemicalfatestudiesof DETA.may presentdifficultieswith

- respectto the polarographicdetectionofanN-nifrosaminederivative of DETAInthesesamples.Shouldthis proveto bethecase,how.ver theAgencynotesthat thethin4ayerchromatographicdetectionsystemswhich.Yordy andAlexander(Ref~.7.andIS) have

developedfor separatingdiethanolamine(a closestructural.analogueof DETA) from N-nitroeodiethanolamnine(a closestructuralanalogueof theN-rtltrosaxninederivative of DE’I’A), andfor detectingN-mtroeodlethanolaminein aqueoussolutions ate detectionlevel of Inanogram/mI(1 ppb) shouldbeadaptablefor usewith DETA and its N-nitrosamine derivative. TheseThin.layerchromatographicmethodshave provento be resistant to Interferenceby.contaminantspresent in environmentalsamplesusedfor chemicalfatestudies.of diethanolamine.In addition,theAgencynotesthat severalinvestigators(Refs.20 and21)havepublishedcomprehensiveproceduresfor thedetectionandquantitationof avarietyofN-nitrosaminesin contaminant-containingenvironmentalsamples,which shouldeasilybe adaptableforusewith respectto theN-nitrosarnlnederivative of DETA.

With respectto thechemicalfat.testingof DETA under anaerobicconditions,the AgencyIsaware thatdenitrificationof thenitrites presentinthe anaerobic,envlronment,thusremovinga necessaryreactantfor theformation of anN-nilrosamiaederivative of DETA. may,indeed,occur.On theother hand, the presenceofheavymetalsor certainpesticidesin theanaerobicenvironmentmay inhibit thedenitrIficatlon process(Raft. 22 through24) so that aN-niirosaminederivative ofDETA may still form. In addition. Bollaget aL (Ref. 25) haveshownthatunfavorable,environmentalgrowthconditions,relatedto temperature. pH,nitrite or nitrateconcentrations,ledtotheaccumulationof nitrite, conditionsconduciveto the formation of theN.nitrosaminederivative of DETA, underanaerobicconditions-inIsolatedculturesof soil bacteria. Thus, theN-nltrosaminederivative of DETA mightwell beproducedfromDETA evenunderanaerobicconditions.However, theAgencybelievesthat the production oftheN-nitrosamnineof DETA observedIn

-. chemicalfate testingofDETA under/ aerobic conditionswould representthe

upper bound for production of thisderivative under less favorableanaerobicconditions.In addition,EPAbelievesthatmostwastewatercontpinIngDETA would be subjectedtosewagetreatmentprocessescontainingat leastoneaerobic step(morefavorable to N-nifrosamineformation)beforereleaseIntowaters whichmightbeusedfor theproductionofdrinkingwater, andthat for treatmentprocessesinvolving both. anaerobicandaerobicsteps,or anaerobicstepsonly, enupper-

bound approximation for the productionof the N.nltrosamlnederivative of DETAfrom DETA couldbe madeby summingthe exp.ectedderivative production at allstepsand assumingaerobic conditionsat all steps.For thesereasons,theAgencywill now requirethe chemicalfate testingof DETA only under aerobicconditions in thefinal PhaseI test rulefor this substance.

In contrast.tothemanufacturersofDETA. theAgencybelievesthat the

- methodologiesdescribedIn thestudiesof Yordy andAlexander(Refs.?and15)andof Pupp(ReL14) andths referencesupon whichhis polarographie methodusbased(Refs. 18and19)do, In fact,togetherwith other studies.(Refs.18.20and 21), constitute a sufficientmodelforthe chemicalfate studiesproposedbyEPA for DETA andarecapableofdistinguishingbetweenthe chethicalandbiological production of an N-nitrosamine derivative of DETA. Thelatter point is now,however,moot,sincetheAgencyhasclarified its positionthat -concernactually existsfor thetotal --production (both chemicaland- -biological)of anN-nitrosanune - -.derivativeof DETAratherthanjuetthi -.biologicalportion of ~production. .. - - -

In summn~y,the Agencyis cofitinuing -to require quantitativechemicalfate.testingofDETA, usingenvironmental -samplesof lake water, sewage,andsoilunder aerobic conditions.In the finalPhaseI testrulefor this chemicalsubstance,andbelievesthat adequate -publishedmethodologiesareavailablewhich,with minor modifications,willpermit the completionof this requiredtestingIn a timely fashion,without theexpenditureof undue time andeffort.F. NecessityforMutogenicity Testiz~.ofDETA -

Many additional commentswere- -receivedonthe proposedTSCAsection4 test rulefor DETA in subject.areuother than the five major Issueswhich - -theAgency raisedfor public commentInthatrule. Commentswerereceivedfromthemajormanufacturersof BETA In thiscountry, Dowand Union Carbide,onthegenemutation andcytogeneticstestingrequiredin theproposedtestrulefor -DETA. Thesemanufacturersbelievethatthere is convincing evidencetodemonstratethat DEFA doesnot havemutagenicpotentialin bacterialormammalian call systems,and,therefore,they feel that the requirementfor genemutation andcytogeneticstestingshouldbe deletedin the final PhaseItestrulefor DETA. . -

Theevidenceto which thesemanufacturersarereferringcoasletsof

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the results of mutarincity. testing. AmesSoimoneilalMicrosomePlateTestsponsorede erby~Dow.orbyUnion was usedwith S typhthuzriwnstralniCarbide, which ~ .ia}v~ftt~dto the TA-1535. TA-1537.TA-i538, TA.-9& andAgency by. thesem,nufae±s~sprior to TA-100. BETA wasalsotestedin thisthe pu’~safinnof ~ proposedtesl’ruie - systemwith theD4 strainof the yeast.for BETA. and~whicharw.-therefore. Saccho.romycescerevisioe.BETA wasdiscussedin theAgency’s testedoverconcentrationsrangingfrom -“Thethylenairiamine Support Document” 0.01to 10.0~slperplaje. both with andfor thatprupoud~sestrule.As described without the additionof a liverIn the support document,thesubmitted microsomalenzymepreparationstudiesconsistoh (1)mntageaicily obtainedfran. theliversof Aroclor-testingperformedby Litton Bioneticsfor pretreated rats. The highestdosetestedDow usingtheAmesSalmonella! - In this study,10.0gil perplate.Microsom.PlateTestand.5strainsof representing9.5mgofBETA. was

- Salmonellatypà/auzthim,aswell asthe selectedbecauseit producedtoxacity.D4strainofSaccb.aramycescarevisice, which wasnotclearlydesudbed.in Swith and without metabolicactivation cerevisia..Litton Bionetics(RaL 4)(ReL 4~,and (2) th. following tests reportedthatBETA wasnegativein allperformedby BuaèyRunResearch test systems,both with andwithoutCenterfor Union Carbide(Ref. 3) for metabolicactivation,andconcludedthateachofthreasamplesof BETA (DETA- BETA wasnon-nuitagenicunderthehighpurit3O BETA-commerciaLand conditionsof the tests.-DETA.hearts cut): (a)An in vitroassay The essentialagreementof thefor specific Iocusmutation(at the negativeresultsobtainedby thetlinghypoxanthine-guanine. . of DETA in the AmesSaimonei]a/

pluosphoriboeyltransferaselecus)in Microsome PlateTestby theNationalChinesehamsterovary(CHO) cells Toxicology Program (Ref. 26)andby

(with and withoutmetabolicactivation); Litton Bionetics (ReL4)calls Into(b) an in vitro assayfor sister-chromatid questiontheearlier reportbyexchange(S~)in CHOcells(withand Hedenstedt(ReL 27) that BETA mightwithout metabolicactivation); and (c) an have demonstrateda directmutagenicin vitro assayof the ability of the test effect in this test system.Hed.nstedtsubstancesto induceunscheduled (Ref. 27) testedDETA in the Amesdeoxyribonucleicacid (DNA) synthesis SolmoneiIa/MicrosomePlateTest using(UDS)in primaryculturesof ret liver SoLo,oneiotj.phthizzriumstrainsTA.-

* cell.. EPAnotesthat only twoof these 1535 andTA—100. both with andwithouttestprocedures,the AmesSaimonella/ theaddition of liver microsomalenzymeMicrosome PlateTestandthe specific preparations obtainedfrom the liversoflocusmutationtestin CHOcells,are ratspretreatedwith ClophenMo, atestsfor genemutation perse.The tests polychiorinatedbiphenyl mixturefor SCEin CHO cellsandfor UDS in similar to Aroclor.BETA demonstratedprimary culturesofrat liver cellsare adirectmutageniceffectin this testusefulasirulicators of geneticdamage, system,whichwas notaffectedby thebut they do not substitutefor essaysof presenceor absenceof the metabolicgenemutationperseandthey do not activation system.However, the BETA

addres. the ability of the testchemicals usedfor the testswas found tobeto inducechromosomalaberrations contaminatedwith unidentified(cytogeruceffects).No cytogenicity test impurities,which theauthor believeddataare currentlyavailable for DETA. mightbe alkylating agents.The author

BETA wasrecentlytestedin the concludedthatBETA, orsomeAmesSairnon.IZa/Mioro.omePlate Teat unidentifiedalkylating ImpuritiesIn the.by theEnvironmentalMatagenasiaTest BETA sample tested.may posea -DevelopmentProgram.-. part of the .nuutagenicand carcinogenichazard.NationalTo ‘olo-Program. awLthe / Basedonevidencefrom thestudiesinvestigatorsconclud.èthatBETA gave- conductedby the National ToxicologynegativetestresultsIn Sal,noruneila Program,(Ref.26) andby LittontyphimrzrnzmstrainsTA-ga TA-lao, Bionetics(Ref.4), the AgencyconcludesTA-1535,,andTA-lw. with or without that thepositiveresults observedItt themetabolicactivation by liver studyby Hedeustedt(Ref. 27)weremicrosorasienzymepreparations probablydueto theunidentifiedobtainedfromArocler-pretreatedratsor Impurities detectedin thesampleofhamsters.BETA was testedina series BETA usedfor testing.-of coacentantionsr~~ngzngfrom 33.ti gig Basedonthe data containedin the -to 10,000j4 perplate(Ref.20).These Litton study (Ref. 4). u well asthedataresults .ss~st~allyegreewith thoes - presentedin theNTP study (Ref. 26), theobservedin a studyperformed~Litton AgencyrnneludasthatDET&hasbeen -Bioneticafor ~ ~ _____ - . demonstratedto be nnrunut~gei~irIn the.Company~RaL4).Zashi.ste4.the -.. bactarium~Salma la.4phimuthtm. -

- -- -

— -~- ~--~------ -—

(strains TA-i535. TA-1537, TA-1538.TA-ga and TA-100), as well asin theyeast. Sacaharomycescarevi.siae(strainD4). under the conditions of theAmesinvitro assay.On the other hand, theAgencymustpoint out thatit doesnotregardthesenegativefindinge withrespectto BETA’s non.mutagenicityto asinglebacteriumandasingle yeastasrepresentativeof BETA’s completemutagemcpotentiaL In ordertoadequamiyassessthe comp’ietemutagenicpotentialofBElA. furthergenemutation andcytogemcitytestingwill barequired. -

Sincethe timewhentheme*agenicitystudiesperformedonDCEA for UnionCarbide Corporation (Ref. 3)were -discussedin theTechnicalSupport --Docnmentfor theproposedteatrule onBETA. theAgencyhas audited,duringFebruary 2810March2.1982.theBusbyRun ResearchCenter in ExportPennsylvama,where thesestudieswereperformed.Therefore,theAgency’scurrentconclusionswith regurdto thesestudieswerereachedfollowing anexamination of the actual raw data f~thesestudiesat thetestfacility.. — --scientific~ betweensiwitlsts..who performedthetestsat th. facilityandEPA staff. aswell asa’Arerlew -.andevaluationof the submitted thaly.reports. -.

BushyRun ResearchCanterhasconductedthvWomutagmuimlystudiesof theability at three samplesof BETA(BETA-highpurity; DETA-commereiáandBETA-hearts cut) to inducespe~clocusmatationsin Chinesehamsterovary (CR0)cells (Ref. 3). In thrpresenceor absenceof a niicrosomalliver enzymepreparation from Aroclor1254-pretreatedmalerats(S-9fraction).the threesamples.ofBETA weretestedin this in vitro systemat concentrationsranging from L25x10 ‘percentto40x 10-2 percent.Theinvestigatorsconcludedthatnoneof the threesamplesof BETA producedspecificlocusmutationsIn CR0cellsundertheexperimentalconditionsemployed.wltior without metabolicactivationby S-afaction.The Agencyagreeswith this- -conclusion,andhas, therefore,determinedthat no furtherin vitro genemutation testingis requiredfor BETA.However,asdiscussedbelow,theAgencyI, requiringin vivagenemutation testingofBETA In this finalPhaseI testrule. -.

BushyRunResearchCenterhasconductedin vitro studiesof theabilitlof three samplesof DET&(DErA-hlgkpurity OETA-cnmm~rciaLmdBETA-heartLcut)to i~,rhie~ajster,.chrosialidexchanges(SCE)In.~in.se.ba~itir.-ovary (CHO) cells..bolh.Iu.tb.pr.s.nc

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and absenceof a rnicrosomalliverenzymepreparation(5-0fraction)obtained from the livers ofAroclor 1254-treated~ I~theabsericeof S- -9 fraction, a statisticallysignificantincreasein SCE wasobservedfar bothDETA’hAgh purity and BETA-commercialatthehighestdosageleveltested(20.Ox10~percent).andasimdarlystatisticallysignificantincreasewasobservedfor BETA-heartscutat the sarondhighestdosageleveltested(1O,Oxlll” percent)underthe

- sameco~litions,An slavat.dthdd”~of SCEwasobservedataDETA-heertscutconcentrationo(20X10”pemeat.buttheincreasewasnct.st.tisticallysIgnificant.In thepresence~ofS-Sfraction, astatisticallysignifiomit.Increaseof SCEwas enonly for the.DETA.heartscutsampleeta dosagelevel011.25x10’peroenL

DowandUnionCarbidecontendthattheseresultsarenot indicativeof apositiverespoesebecausetheyocceratsingledosagelevelsandno dose-responserelationshipscanbedemonstrated.The Agencydoesnotagreewith thiscontention.The OTS testguidelinefor thein vitro SCEassay(Ref..28)states:~‘Thereareseveralcriteria fordetermining apositive result oneofwhich is a statistically significantdose-relatedincreasein thenumberof SCE’s.Mother criterion may be basedupondetectionofareproducibleandstatistically significantpositive responsefoi at leastoneof the test substanceconcentrations.”SinceSlesinskici a!.‘(Ref. 3) performedonly oneexperimentfor eachof threeDE’I’A samplesin theabsenceof themetabolic activationsystemfor SCEin CR0 cells,aswell asoneexperiment foreachsample in thepresenceof themetabolicactivationsystem,nothingcanbesaidregardingthe exactreproducibilityof thefourstatistically significantpositiveresultswhichoccurred at single dosagelevels.However,sincestatisticallys%nifi&~antIncreasesof SCEwereobservedInseparateexperimentswith thethree -BETA samplesIntheabsenceofS-~

- fractionat concentrationsranging from’10.0x10’~parceidtomoxio’ percent.theAgencyconcludesthatBETA(constitutingthemajorfractionofallthreesamplestested)hasexhibitedapositiveresponseIn this in vitro system.Therefore,no furtherin vitro testingofDETA for SCE in mammaliancallsisnecessary,but,asdis~m~edbelow,theAgencyis requirlnginvivogaas--mutationtestingof DETA In DrosophilamR!nni~gnIMi~.based.in.part,on BETAspositiveresponsein the~. vitro testfo~

- sCEIncRaceiis.

With respectto the studiesbySlesinaldataL (Ref.3) on the ability ofthreesamplesof BETA (DETA-highpurlty DETA-commercialand BETA-heartscut) to induce unscheduledDNAsynthesis(UDS) in primaryculturesof

- rat hepatocytes,the rangeof dosestestedwasinappropriatelyselected.This doserangewasselectedon thebasisof toxic effectselicitedby BETAin CR0cells In the cours.of studiesoftheability of BLTA to elicit specificlocusmutationsIn thesecells.Thedose.rangeselectedfor LIDS testingshouldhavebeenselectedon thebasisofDETA’s toxicity to the primaryculturesof rat hepatocytesusedfor thsLIDSstudies.not on the toxicity observedforBETA in CR0cells.In addition,thereisno indication in theresultspresentedforthe LIDS studiesthat the highestdoseofDETA tested(1~0x1O”percent)was,infact,toxic to th. primaryculturesof rathepatocytesusedfor this test,asisrecommendedinboth theOTS testguideline(Ref.28)andthe Gene-TonWork Groupreporton this assay(Ref.29). Nonetheless,statisticallyaigniflr~~ntincreasesIn UDS were observedbothwith BETA-highpurity andBETA-heartscutat concentrationsof0.01x10’ percentand0.3X10’percent(basedon DNA-bound radioacthnty3.but neither sampleof BETA elicitedstatisticallysignificantly elevatedUDSat aconcentrationof 0.1x10”1 percentDETA-high purity induced an elevated.UDSat o.ixio’ percent,but theincreasewasnot statistically significantNosignificanteffectswere observedforBETA-commercialIn this testsystem.

The Agencydisagreeswith Dow andUnion Carbide that. becausethestatisticallysignificantpositiveresposesIn the LIDS assaysof BETA couldnotdearlybeshownto bedose-related.theresponsesobservedwerenot trulypositive.The OTS testguidelinefor thein vitro liDS test(Ref. 28)containsastatementto the effectthatareproducible,statisticallysignificantrespons.with respectto UDS for atleastonsdosagelevel maybean -Indication of a positiveresponseIn thistestsystem.No attemptwasmadebySlesinsklat a!. (Ref. 3) to verify thereproducibilityof theresultsof theUDSstudies.However,becausetwo of thethreeDETA samples~(DETA.highpurityandBETA-heartscut)eachgavestatisticallysignificantpositiveresponseswith respecttoUDS Inseparateexperimentsat concentrationsof 0.01xiO” percentandO.3xlr’percent.theAgencybelieves—thatBETAmustberegardedasanin vitro ~of UDS in rat hepatocytes.Thefact4hat

- BETA-commercial(whichIi lesspure -

thaneitherDETA-heartscutorBETA-high purity) failed to give astatistically—significant elevationofliDS in this testsystemmaybe relatedto inhibitoryeffectsof impurities presentin thissampleof BETA. TheconclusionthatDETAelicits UDS in primaryculturesof —rat hepatocytes.togetherwith theconclusionthat BETA inducesSCEinCR0cellscultured in viti’o, indicatethatBETA shouldbetestedfor its ability toinducein vivagenemutationsinDrosophilamelo.aogaster.asdiscussedbelow. -

In summary, theAgencycamsotadequatelyuses.themutagrmcpotentialof DETA with the informationcurrentlyavailable,BETA ha. beenshoasafReEs.4and 20) to be.- - -nonmetagenisto five sirainsof thebacterium,Salmonellai4imer,um,- ‘andto the D4strain of theyeast.Socthammyes,carey/sloe,undertheconditions of theAmesSalrr,oneila/ - -Microsom,PIateTestIn addition.~tudie~exist (Ref. 3)which indicats-thatDETA do,snet-Inducesped&toes.’mutations in Chinesehamste’overy(CR0)cells,but doesinduce~sister--chrorn.tidexchange(SCE)In’OfO’celIs.enddoesinducennscheduledDNAsynthesis(liDS) In primarycnlt~e,ofrat hepatocytes.Ontheotherhand,theAgencynotesthatnocytogenicitytestdata are currently-availableforBETA.TheAgencybelievesthatthepo5ttiveresponseswhich DETA has exhibitedIntheSCEand liDS mutagenfcftyassaysindicate thatthis substancemayposeanunreasonabli risk of bothgenemutations and cln’omosomalaberrations.notwithstzndIng~the. -negativeresultsexhibited by thissubstanceIn in vitro genemutationassaysin bacteria,yeast,andmammaliancell. in culture-,Therefore.the Agencyis requiring bothin vivagene mutation andin vitro andin vivacytogenicitytestingof BETA hr thefinalPhaseI testrulefor thIssubstance.

The AgencyIsrequiringthatBETA betestedfor in vivagenemutation effects.utiH2ilig the-sex4lnkedrecessivelethalassayin Drosophilameianogaster.If theresultsare negativefor BETA in thetestin Drosophila,no furtherin vivagenemutationtestingIs required.If theresults arepositivefor BETA In theDrosophilasystem,thentheAgencyisrequiringthat BETA be testedIn. themousespecificlocusassay.Guidelinesfor all of thesetestprocedureshavebeenpublishedby theOfficeof Toxic -Substances(Ref.28). EPA is alsorequiringthatBETA be testedfor its- -

- ability to induceboth in vitro.aidin-. vivor~Pnn1wJ~!tAJaberrations, usingthe(

recessivelethal assayin Drosophila, mousespecific locus test and theandthis requiremenriscontainedin the heritable translocation test), EPA hasfinal PhaseI test rule for DETA. decidedto utilize automatic triggers

With respectto cytogenicitytesting, if betweenassayscontainedin Iower,tlera substancegivesa negativeresponsein tests,and a “presumptive automatican in vitro cytogeneticaassay,It is then thgger andopt-out” approach betweentestedin an in vivocytogeneticsassay. lower-tier testsand end.pointtestsinIf thesubstanceexhibits a negative . this final PhaseI-test rule for DETA.responsein the latterassaysystem,then Under this approach,EPA Isno furthercytoganicitytesting is promulgating a tieredtesting schemeforrequiiet If asubstanceexhibitst mutagenicityfor DETA with automaticpositiveresponsein either the iii vitro or triggersto additionalesutagenicity - -in v/va cytogeneticsassay,then the testing(Inciliding thetwo end-point - -substanceis testedhr thedo,nfrt~nt tests).Befor.testingis initIatedin one -lethalassay.A positive responseIn the or both of~the end.pointmutagenicltydominantlethaiassayindicatesthatthe tests.EPAwill hold apublic-programsubstanceshould be testedIn the ~ If theresultsof theprevioustierheritable traaslocationassay.A - testsarepositive.Public-p.rtic2pationIn.negativeresponseinelther of the latter thispro~amreview will be’either In thetwo assay.indicates thatno further form of writtenpublic comments~acytogenicitytesting Is requiredfor the public meeting.Requestfor publicsubstance.Sinceno cytogenicitytest . commentsor notificationofapublicdata areavailablefor DETA. the final meetingwill bepublishedin theFedsialPhaseI testrule for this substance Register.If, after the reviewof publicrequires thatBETA be testedIn - comments,no changeIn the test.accordancewith the tiered testing programIs deemednecessaryby ffl’A.sequencesfor bothin vitro and in viva testingwill continueto th.next tsat~cytogenetics.asdescribedabove.The without delay.EPA will provide-Agency’sresponsesto commentsoe.the - notification to the test sponsor($that:tieredtestingsequencesfor gene thenext tier-test-shouldbeconduct,d11..mutation testing andfor cytogenetics theAgencybelievesadditionaltestIngIstestingmaybefoundrntheflnalPhasel nolongerwerrantedaseresultofthetest rule for the C, aromatic- - earliertestresults,public comment,hydrocarbonfraction (50FR20682). scientificjud~nent,andother

Asdescribedin detailin the final appropriate factors,EPA will issueaPhaseI testrule for the C,aromatic proposedamendmentto “opt-out” by~~hydrocarbon fraction (50 FR20662).the repealing the existingrequirementand.Agencyfeels that there Is a consensusin after consideration of public commentsthe scientificcommunity onboth the on theproposedamendment.issueaneedfor, andthemanner of, identifying final decisionwhether to rescindthetoamnialianmutagens,andthat its - rule requirement.This approachoffersproposedschemefor Identifying these theadvantageof allowing theagentsis inkeepingwith’those incorporationof scientificjudgment -.recommendedby expertsin the field of ontheweight of the evidencemammalian mntagenesis.Further,-while ~- after’ the initial testingtiershavebeenIt I. recognizedthat thereis, asyetno completed-andallowing changein lestgenerally acceptedsingle methodology requirementsto respond to specificfor estimatinghuman risk from- chemicalissues,while i~otsignificantlymutagenicagents,it is the Agency, - delaying-higher-tier testingwhen it isview thatappropriate methodologiesdo deemednecessary.exist and areusable.Therefore.the- EPA’has decidednot to usethepublicAgencyconcludesthat it Isappropriate programreviewapproach betweentheat this time to obtain mutagenicitydata . lower.tler mutagenicitytestsfor the . -.~,nDETA with whichto perform BETA test rule. EPAbeltoves~ useofestimatesof mutagenicrisk for this automatic triggersbetweenthesetiers Issubstancefor regulatory use,should suitable. It shouldbe notedthat thisDETA prove to be amammaliangerm- doesnotexcludethe publicfroni’cell mutagen. - . requestingmodifications-inthe.test -

For reasonsmorefully describedIn program. Provisionsare availablewiderthe final PhaseItest ruleforihe C. - section21 ofTSCA for the public toaromatic hydrocarbon fraction (50FR petition EPAat anytime to amendanile20682),EPAbelievesthat theuseof undersection4.EPA. Test Rule -automatic triggersbetweenthe assays - DevelopmentandExemptionProcedurecontainedin themutagenicltytesting’ t~uIe.published In the FsdiraI Registerofschemefor BETA-Is appropriate: October10.1984-(49FR 39774).Includes-however, in en effort to Incorporate proceduresfor-Industrytests~Cnsor~toscientific judgementpriOr to thi useof requesticodificafionsto ‘testguidelfnes

the end-point mutagenicitytests(L.., the (but notto testrequirements).”

/

21406 - FederalRegisterI Vol. 50. No. too / Thursday, May 23, 1985 / Rules and Regulations

(test sequencesoutlinedin Unit IV.B. ofthis final PhaseI testruM.~ . - -.

The generaisequencàoftieredtestsusuallyemployedby EPAinassessingthe mutagenic(both.genemutation and.cytogenetic)potentialof:ch~mfcalsubstances,portionsof whIch arerequired in this final PhaseI test rule forDETA (seeUnit IV.B.). have-beenpreviously describedin proi5~isedtest

- - rules issuedby the Agency for meóityloxide (48FR 30699) cresols(48 FR21822),andethyltoluenes,trimethylbenzenes.and Cm arom8tichydrocarbon fraction(43 FR23068),andare morecompletelydescribedIn thefinal PhaseI test rulefor C, aromatichydrocarbonfraction(50FR 208621May17, 1985).Although thesegeneraltestsequencesareusuallyemployed,theAgencyultimately specifiesthe requiredmutagenicitytest for eachspecificche~calsubstanceon acase-by-casebasis, With respectto genemutationtesting, if asubstancetestsnegativelyinthe genemutationassayin Salmonella.It is then testedin the specific locusmutation assay.In mammaliancells Inculture. If the substancetestsnegativelyIn the. latter assay,thenno further genemutation testingisrequired (in theabsenceof other positive mutagenicitytest data). If the substancetestspositively in either theSalmonellaassayor thespecific locusmutation assayinmammaliancells in culture, then it istestedin thesex-linkedrecessivelethalassayinDrosophila.If the substanceyieldspositive resultsin the.Drosophiaassay,then it is testedin.the mousespecificlocusassay.NegativeresultsIn

• either theDrosophilaassayor themousespecificlocusassayIndicatenofurtherrequirementsfor gene-mutationtesting.

DETA testednegatively,both withand withoutmetabolicactivation, InAmesassaysusingbothSalmonellatyphimuriumandSacchoxomycescarevia/ce.and alsotestednegativelyInaspecificlocusmutationassayIn’Chinesehamsterovarycilia In culture.BETA would. therefore,normally not be

.ubject,Intbe.bsesceo!otherpositlvemutagenicitytestdata,to. requirementfor furthergenemutation testing.However,aspreviouslydiscussed.theAgencyha. concludedthatDETA

- Inducessister-chromatidexchanges(SCE) in Chinesehamsterovary-cells inculture and inducesunscheduledDNAsynthesis(UDS)in primary culturesofrathepatocytes.Thepositive resultsdisplayedby BETA In. thesetatter twoassaysindicateto the’AgencythatBETA shouldreenterthe tieredtestsequencefor gene‘mutationtestingatthesecond-tiarlevef,’thesex-linked

Federl Register,L Vol. 50, Ne. 100/ Thursday.May 23. -1~85/ RWer and Regulations 21407

Sincethe time at which the proposedtest rule for BETA waspublishedby theAgency (47FR 183861April 29.1982).theEPA hasadopted anapproach ofrequiring tiered testingsequencesforboth genemutation and cytogeneticz-testing which containautomatic triggersfor requiredchroniconcogenicitytestii~when a chemicalsubstanceelicitspositive test resultsin certainof themutagenicity assays.The mutagenicitytesting actually requiredfor a givenchemicalsubstance.aswell as the.selectionof thosemutagenicityassays(if any)for which positiveresultswilltriggeran automatic requirement forchroniconcogenicitytesting.is -determinedon acase-by-casebasis.Following carefulevaluation, theAgencyhasconcludedthatsuch-triggersfor chroniconcogenicitytestingare -appropriate for DETA. Becausetheproposedtestrule for DEI’A containedno requirement for onccgenicitytesting.either as an absoluterequirement or asa result-of positive testresultsinspecifiedrequiredmutagenicity assays~EPA is proposingelsewherein this issueof the FederalRegisterunder section -4(a) of TSCA. that manufactweieendprocessorsof DETA bereqtared to - -conduct chroniconcogenicitybioassaysof this chemicalsubstance,if positivetest results areobtained for D~TAinany of the following mutagenicityassaysrequired for this chemicalin the finalPhaseI test rulefor DETA (1)-The sex-linked recessivelethal genemutationassayin DrosophilamelanogasLer.(2)the in vitro cytogeneticsassay,or (3) thein vivocytogeneticsassay.

C. RoleofProcessorsin theTesting-ofDETA

Many commentswere recei~&withrespectto the responsibilitiesandobligations of processors.both withregardto thespecificproposedtestrulefor DETA andwith regardto theTSCAsection4TestRule andExemptionProcedures.In general.Thesecommentswere consideredand-addressedIn thefinal rule onTestRuleDevelopmentandExemption Procedures.publishedIn the,-FederalRegisteronOctober10,1984(49

- FR.39774). - - -H. RequirementofStudyProtocolsRatherThan GeneralStudyPlans

Another issuerelatingto the proposedtestrule for DETA. which wascommentedupon -by CMA andAlliedCorporation(Allied) is the Agencysintentibn to publish proposedstudyplans submittedfor DETA for public

comment arid to incorporatethe.sp~ciiãc generalpopulationto asuspectt... ~ iy,difled SY ~m-uiuiogefl.N-Nltru5udietttunolamine. a

known animal carcinogen(Ref~8through 10). is formedunderenvironmental conditionsin waters

- coatais~diedianolamine,aclosestructural analogueof DE’I’A (Ref. 7).Chemical fate testing of DETA isrequiredto det”4”lf~a.~.nitr-osaminederivative of DETA canbeformedunder environn~s~talconditions.The finding of potential-adversehealtheffectsasa resultof subd~ronicorchronicexposureto DETA is based,inpart. upon thestuRe,-ofF8~ino(Ref. 2),which indicateDETA-related adverse

L CovfideótiollnforrnationCorned iii -effectsonthe liver, lung~andkidneysStudyProtocols - - - (and. possibly,the spleenandadrenals)

CommentsfromAllied on the ~ ~onica~y expo~dtoBETA.proposedrule for DETA were~ This finding is alsobased.In part. uponby the Agencywhich raisedthat firms the studiesof TrubkoaedT.~tyakovaconcern about the possiblebreach of the (Ref. 30). whiciidemo~e1ethat

- confidentiality of proprietary exposureof rahintafor8 months toinformattonwhich might occur in Phase DETA via the thinking watercan resultII of the testruledevelopmentprocess. ~ ~ ,~gn1ficamdeceasein prothrombinin general,throughthe negotiationand activity and significantIncreasesin the -publication of detailedprotacolalea - -- euivitisz.fs~n giutaru~ate-.oxalate

- requl dta.ta4oeBETA e,~orethe,~ nüramiiei~naglutami~Te-pyruvatesubstanceswhicharethesubjectsof - ~nsaminase The Agency~ ---differentTSCA section4 teststiles.. that there are insufficieht ~

- .,Allieds-corame~were-cvnsfdered~à’:~*~enee i!dch th~’effndi1mfth~addressedin the final ruleon TesR~~manufacture,distributionIn~ie’ce.Developmentand Exemption - - ~ use,or disposalof DVM erProcedures,published in theFederal any combination of suchactlvities.onRegisteron October10. 1984 (49 FR - hwnanhealth canreasonablybe ,.~, -39774). - ~‘ determinedor predicted.and thattesting‘lIV. Final TestRule for - - of DETA with respectto sucheffectsisDiethylaeetriamine - necessaryto developsuchdate~ - -

A. FindingsThe EPA finds that the manufacture.

processing.use, and disposalof DETkmaypresentan unreasonablerisk of -injury to human helath due to potential-

- mutagenic. oncogen~.and.subchroniceffectsof the substancefor the reasønspresentedin the proposedtestruleforBETA (47FR 183861April 29. 1982)andmore fully describedin the support’documentprepared for that pmpoaed’~rule.The finding of potential mutagenicrisk is basedon thestudiesof Slestuskietci. (Ref. 3), which indicite that BEtAinducessister-chromatid exchanges -(SCE) in CHO cells In cultureand -inducesunscheduledDNA synthesis(UDS) in primary culturesof rathepatocytes.The finding of potentialoncogemcrisk is basedonthehypothesisthat aN.mtrosamlne -derivative of DETA may be formed Inenvironmentalwaters,soils,and

- sewage.and may survive the treatmentof contaminatedwatereprior to their -use for drinking water, thus exposingthe

a re~n1tof public comments)into thermal PhaseII test rulefor DETA asenforceabletesting requirements. Anymod ficadna.oLthasa.raquAtemIUtswould requirethe Agency~sapprovaland,whenappropriate, publication ofthe proposedmodifications-forpubliccomment. Commentsfrom these firmswere consideredand addressedin thefinal ruleonTest Rule DevelopmentandExemption Procedures.publishedin theFederal Registeron October10,1984(49-FR-39774).

(

(

B. RequiredTesting ,.“The EPA is requimipg’~atBETA be

- testedfor oral subefironic(90-day) -- toxicity in aLleestonenmmmalianspecies.~i accordancewith theOTSHe~EffectsTest Guidelines,

- publishedby the National TechnicalInformationService(NTIS.P982—232984),with respectto oralsubchronictoxicity studies,testingof DETA in atleastonemammalian specieswill beconsideredsufficient,asopposedto therequirementfor testingin at least twomammalian species,aspresentedin the’proposedtest rulefor BETA.

The Agencyis requiringthat a dermalabsorption study be conductedwithBETA in the samemammalianspeciesselectedfor oral subchronic(90-day)testing. -‘

The-EPA is requiringthat BETA betestedfor mutageniceffects,bothwithgenemutation andcytogeneticstesting.and is requiringthe following sequencefor this testing.IWNO ccci SMOISM

/~*

21408 FederalRegister/ Vol. 50. No. 100/ Thursday.May 23. 1985 I Rulesand Regulatloril

-

Testir~of ~TA for Ir~ucir~ In Vivo Gere ~tztatioRs

Sex-linkedrec~sivelethal. —assayin - -Drosophila

-- -

Positiv -

specific~ -1OQJS assay -

~~Negative — ~—~b further- tastir~

Testirs~of ~ for Inducir~O’ir~oson~a1AbetTatior~

Datiinantlethal

assay

- - - - -- - Positive

ccci

~rit~letranslocationassay -

(.

assay

~sit ive

In vitro - ‘ - ‘ l~further - -cyto- —~ Negative —~genetics- -

In vivo cyto--geretics - -assay -

—~ Negative ~ testirç- ‘. -- -...

-

-~-

-

- - - - -

-- -

:

P:~sit ive

—-s--- :~tj~ -

FederalRegister / Vol 50.No. 100 / Thursday, May. 23..19651 Rules and Regulatione.~. 2140g

The Office of Toxic Substanceshaspreviouslyissuedguidelinesfor all of -the test methodsmentionedabove(Ref.

-23). - — - -- - - - -The AgencyIs also requiringchemical

fate testingofBETA In environmentalsamplesof soil, lakewater;andsewage.underaerobic conditions,following thegeneralmethodologyutilizedby YordyandAlexander(Refs.7 and15)andTateandAlexander(ReL18). The final - -requirement-foechemicalfate testingofDETA has changedfrom that appearingIn the proposed--testrule-foeBETA inthat noanaerobicchemicalfate testing-Is now required. - — • : -

CTessSabstance ““ -EPA is requiring thatarelativelypure

grad.ofBETA beused.,thetestsubstance.A purity of atleast99percentis specifiedIn this rule. BETA ofthis purity (DETA.High Purity) iscommerciallyavailable.D. PersonsRequiredto Test

Section4(b)(3)(B)specifiesthat theactivities for whichtheAtlminiatratormakessection4(a)findings(manufacture,processing,distribution.useand/ordisposal) determinewhobearsthe responsibility for testing.Manufacturersarerequired to testif thefindingsarebasedonmanufacturing.distribution, use,or disposaL“Manufacture”is definedIn section3(7)of TSCA to Include “import.” Processors-are requiredto test if the findings arebasedon processing,distribution,use,ordisposal. -

BecauseindustrIalworkers.consumers,and thegeneralpopulationmay beexposedto DETAduringmanufacture,processing,useanddisposal,EPA Isrequiringthat personswho manufactureorprocessor whointend to manufactureor processthischemicalfrom the effectivedate-ofthistestruleto the endof the reimbursementperiodbesubject to therule.TheendofthereimbursementperIod.will be5yearsaftarthsd.a~1nefor submittingthelast final report underthePhaseIItestrule.AsdiscussedIn theAgency’s --feetrule developme~-aadexemption /procedures(40 CFRPart790),EPAexpectsthatmanufacturerswill conducttestingandthat processorswillordinarily beexemptedfromtesting.

BecauseTSCA containsprovisions toavoid duplicative testing,not every

personsubjectto thisrulemustindividuallyconducttesting.Section4(b)(3)(A)of TSCA provides thatEPAmaypermittwo or moremanufacturersor processorswho aresubjectto theruleto designateonesuchperson-or aqualifiedthird personto conduct thetestsandsubmitdataontheir behalL

Section4(c) provides-thatany personrequired to test may apply to EPA for anexemptionfrom that requirement. EPA’sfinal regulationsfor the issuanceofexemptionsfrom testingrequirementsarein40CFR Part790.In accordancewith theseregulations~anymanufacturer or processorsubjectto aPhase!testrulemaysubmit anapplication to EPA for an exempfloe..-from submitting itudyplansandfromconductinganyorill ofth. testrequiredundersuchaavis.Ifmanufacturersperformall therequiredtesting.processorswill begrantedexemptionsautomaticallywithout- :havingto file applications. -Manu1aa~-~andprocessorswhoare-subjectto th, testing requfrements of -this rulemustcomply with thetestruledevelopmentandexemptionproceduresin4OCFRPart7gO. -

EPAIsnot requiring the submissionofequivalencedata asa condition forexemptionfrom therequiredtesting.Asnotedabove.EPA is Interestedinevaluating theeffectsatutbutable toDETA itself, andhasspecifieda - -relativelypuregradesubstancefortesting.£ TestRuleDevelopment -

Developmentof this testrule-forDETA will be atwo-phaseprocess,InPhaseI. this test rule isbeingpromulgatedfor BETA specifyingcertanhealth effectsandenvironmentalfatecharacteristicsfor whichtest dataare to be developed.In PhaseII,following promulgationof the Phase!test rule, thosepersonssubject to therulewill berequiredto developstudyplansfor thedevelopmentof datapertaining to the effectsandcharacteristicsspecifiedIn thePhaseIrule.Within 30days from the effectivedate of the final PhaseI test rule,manufacturersmustsubmit toEPA aletter stating their intention to sponsortesting or an application for exemption.Testsponsorsmustsubmittheir study

- plansto EPAwithin 90 daysfromtheeffectivedateof the PhaseI testrule.Afteranopportunityfor publiccommentEPA will promulgatearuleadopting the study plans,u-proposedormodified,asthe teststandardsandschedulesfor BETA for thetestsrequiredby the PhaseI rule, Testing willalso be subjectto EPA’s genericTSCAGLPstandards.Personswhosubmitthestudy planswill beobligatedto perform-the testsIn accordancewith theteststandards and schedulesdeveloped.Modificationsto the adoptedstudypla~scanbe madeonly with EPAapproval. - -

Processorswill not berequiredtosubmitletters of intentexemption

applications,and study plans,andtoconducttesting.unlessmanufacturersfail to sponsorthe requiredtests.Thebasis for this decisionIs that -manufacturers areexpectedto Indirectlypassthecostsof testingon to processorsthrough any price increaseof BETA.F. Reportfr~gRequirements

EPAisrequirlngthatalldata -developedunder this rulebe reported inaccordanc,with theTSCAGood-LaboratoryPractice(GLP)-standards

- which were publishedIn 40CFR.Part792.Thesefinal- GIPstandardsapply tothisrule. - - -- EPA Is required by TSCA section

4(b)(1)(C)to specifythe timeperiod• duringwhichpersonssubjectto atestrulemustsubmittestdata.Thesedeadlineswill beestablishedIn thesecondphaseof this rulemakingInwhichstudyplansareapproved.Theproceduresfor the secondphase - -

- rulemakingaredescribedin 40 (~RPart790.

‘~SCAsection12(b)requireithatpersonswho exportor Inten~b~ert

- to aforeigncountryachemicglsubstanc,or mixturefor whith~submissionof datais requiredunder.

- section4. suchas BETA,notlfy EPAofsuchexportationor intent to export.While the results of requiredtestingmaynot be availablefor sometime, a noticeto the foreign government*bout theexport of BETA servesto alert them totheAgency’sconcernaboutthesubstance.It givesthesegovernmentsthe opportunityto requestsuchdat&thatthe Agencymaycurrentlypossessonthesubstance,plus whateverdata maybecomeavailableasa resultof testi~activities.Thus,upon the effectivedateof this rule, personswho exportorintend to export BETA mustsubnd~-noticesto theAgencypursuantto TSCAsection12(b)(1).For additional -Information,see49FR 45581(November19,1984).

TSCA sectIon14(b)governsAgencydisclosureof all testdata submittedpursuantto section4 of TSCA. Uponreceiptof data required by this rule. theAgencywill publishanoticeofreceiptin the FederalRegisterasrequiredbysection4(d).Testdata receivedpursuantto this rulewill be madeavailable forpublic Inspectionby anyperson. exceptIn thosecaseswhere the Agencydeterminesthat èonfldential treatment-must be accordedpursuantto section14(b) of TSCA. - - - -C. fnfonementProvisions - -

The Agencyconsiders-failuretocomplywith-anyaspect-of a section4 - -rule to be a-violation of section15of

~

- 214i.~ FederalRegisterI VoL 50, No. ~o0 / -Thureday. May 23. 1985 /— Rules und &e~ilañ~

, TSCA.Sectioa13(1)of TSCA makesit- - - unlawful for anypersonto fail or refuse-- - tocywithany~eorc~iuued

- - ‘uñdcl~ection4.Sa~fon15t3la1TSCA-makes it unlawfulfor anypersento Sail~.r refuseto: (1 ablle~ormaintain

- - te~o~~)submitrepo~.a&fces,or- - - other information,or (3)permitaccesstQ

- - orcopyingof recordsrrqui~edby theActor sayregulationissuedunderTWA

- - Additionally. TSCAsectico15(4t- - - - -- makesit uilawful for any person.to fail

-- ~-orrefusetoper~tentryor inspecti~’oas- ~requiredbyse~~4~”ii.Section11 -

applie,to any‘establishment,facility.- or other premisesin whichchemical

substancesor mixtures are- ; manufactured.processed.stored.or held

‘~beforeoraftertheirdistributionin- commerce.,. ,“Tbe Agencycona~en

atestingfacility to beaplacewherethechemicalisheld or storedand.therefore.subjectto inspection.

- - -- -- Laborctory audftslinspecdonswill beconductedperiodically in accordancewith the ~rocedvresoutlinedin TSCA.section11 by da~natedrepresentatives

of the EPA for thepurposeofdetermining compliancewith thefinal

- rulefor BETA. Theseinspectionsmay-- becon~Zedfor purposeswhich

Includeverificationthattestinghasbegun,that schedulesarebeingmet.- -thatrapbrtsaccuratelyreflectthe

- - under~~raw data andinterpretationsand evaluationsthereofandthat the

~ studiesarebe~conductedaccording- -- to theTSCAGLP standardsandthetest

- - - - - -- standardsestablishedin thesecond- - phaseof this rulemaking. —

-- - - EPA’s authorityto inspectatestingfacility alsoderives-from section4(b)(1)

- ofTSCA.wbichdlr,ctsEPA topromulgate standardsfur the -

- -- developmentoftestdata. Thesestandárdaansdefledins.nd~3~12J(B~l

of TSCA to Include thoserequirement.necessaryto sui~ethat data developed-undertestingrulesarereliabl, and

- adequate.and suchotherrequfremsuisas areaeceumytopioetdsmeh

— assurance-The A~cymafatafesthatlaboratoryinspectionsarenecoemrytoprovidethis usursncs.

Violatorsof TSCP~are subject tocriminal and civil liability. Personawho

submit materfaflymisleadingorfalse- InformationIn connectionwith be

requfre~entof any provision of this rulemay be subject topenaltiescalculated

- asif they hadneversubmittedtheir— - dath. Under thepenaltypr~ioaof

sectionis ofTSCA. anypersonwho- violatessection iScorid-besubjectto a

civil penaltyof up to $25.DO0perda~foreachviolatIon. r~tanffo~ofviolationscouldleadto the impositlønatorfminaI

penaltiesof up to 125LQ00for sach.day of On the basisof this study, theA~ncyviolation and imprisonmentfor up to - - believesthat therewill be availabletestoneyear.Otherremediesareavailable facilities, andpersormelto pezfor~theto EPA undersections7 end17of TSCA. testing requiredin this test rule.suchasseeking1n injunctionto restrain --violationa-oITSCAs.ction4 - em -

Individuals asweltascorporations EPA has establishedapuk~Acrecordcould be subject to ~ ~ for tins rulamakEig (dochatnumberSections15sadi&o1TSCAappl~te OFrS-42012B)u~hichisavad~Iceany person who vio&~tesvarrouc uaspectmo.is. ikeOP7SliesdtngRoom.

provisionsof TSCA~.EP~~y, ~ Jtm E~-1W.4t~1.M StrestSW..discretion,pcocwi~ ~ Washiogtoti.D.C..from& a.m.to4pm..aswell ascomp*o~es.themseises.In ‘-Monday throughFriday..ureØ legalparticular,this ixhulesinàvidaslswbá -thbliayi.Thnr~sdInchei besócreport falseinformationor ~ ~ mtutma-bcotheAgeuq~ usto be reported. heSiMS~’Is.the - -.: developingthi,proposal,and,ubnusmonof f~Jss.~ or a~propnateFnderslRegisternotices.frau~,otstatements u~icn_co l’heA3encywill supplementtherecordunderisU.S.C..1001. - - - with~ddiba~alEforusetion—~Is, ~ receiveLThs-g~dsow the

V. ~ Analymaolivia foilowi~~g -To assessthepotential ec~- ~ ~ r~..~

of tbmfmat PhasuI test .~ . -- ____ _____EPAhaspreparedan e~oeunc - (1) FederalRegisteraofl~pwiaiute,evalso~that ~ ~ of -~tothis rule.consistingoftthe requiredtestingcodcoutyzes-lcor - - :(~)Noticeof final rule on BETA.iuarke*~ t~~ jb) ~‘~~v~”p”4 ruleonE~TA(1) Demand-s j2)cost ~ (4~’FR1~00J. - -charactenatico.(3)~mi~ysti~ture.- -~ - (u)Nolicecontaining thefTC -and(4) marketexpectations -~ desipietionofBETA to thePriority List

Basedon a total ~ costof - (48Ffl 2J138~.$220,400to$487,200andanannualized‘- - (dt Noticeof !nal rule or EPASTSCAcostof $57i15to $t~,ZS3.the~umic GoodLaboratoryThncticeStandards(41evaluationofDETA indicates thatthe FR539~). -potential for ~ economiceffects (ci NoticeofSnalruleontestruledueto the~etheatedtestingcosts.isto~~devnlopnieiitand .iou proceduresThis conclasion Isbasedonthe --~-- (49FR3~774i. -following obserratlon~til Thedemand (fl NoticeoflanErulecon~1~nfngdatafor BETA is relatively inelastic dueto -- reimbursement(48 FR317551. -limitedpotentialforsubstitutionin and (2) Support documents,consistingo~uses.(2) themarket expectationsfc~ - (a)DiethylenatriamionsupportBETA aregenerally favorvbin~and (37 document.the relativemagnitudeofthetestco.Us (b) Ecovo~napactana3ysiaoLlisoiminor, I.e., anesthuatad0.49cnt per test rule for BETA. -poundto theupperhumid.This (3) Commusi~u~conmati~otrepresents0.31percentofthesafesprice (a)Written pebhe~of BETA. The economicanalysis - - - (l~3SemaarIs~of telephone -presentingtheseconclns’n~~is inthded~--coi.~svstioei,In the pubtic record for thisrulemaking. ~ M~s~gs~*snVI- ~ of~ ~ (d) Repode—çabI*sbedaridPursc& - unpualmbadlactonimaterials,ki~~

on )~1)a requires EPA (4)Testprotocalfin-s .4——Ito consider~be a ions ~ absor’~.taèv ofavailability oldie facilitiesand ---- -personnelneededto performthetestIng ~8•Referencesrequiredunder thenil..” ~efore. EPA- :~1i);N~H,Natisndie~,for

-- conductedastudyto assessthe ~ ~ sassy~g _____availability of testfacillliss and -‘~-- -- - Occupational--HasardSarsayDataBus.personnelto harxfle the ~d.4DkmiI Washington.D.C. U.S.Depsr~ititH..hkdemandfor tastingservicescreatedby EducationaniWelinw.~..section4 testrulesandtint programa (2) Fuilno.M. “~icperhiieiitaIStiuti.son th

of __ __

emi , - esting ustry Prolile of Itos.fI’ransmint byToxicologicalTesting, October.1981,. - Servicefor D~Ccan be obtainedthrough,theN~Sunder - -- ~j sti~ai.p.s,-G~&.~sw..C~ -publication nuinber PB$2.-44lll73.~ - P.J..anéHm W.C. “Uis*hy~mme

FederalRegister/ Vol. 50, No. 100 / Thursday,. May 23. 1985 I Rules and Regulations 21411-

wit jo MutagenesisStudies: 3-TestBattery.” Determinationof SecondaryAmines.” Z.Projectreports43-90.43-na.and43-120. Anal. Chen.185.161-174.1962.(Translated

- - Bushy RunResearchCanter.19t0.(Subieltted lot EPA by Scitran).to Union CarbideCorporation). - (19)Clang,S.V.. andHarrington.G.W.

(4) LItton Blonetics,-Inc. “Mutagenicity “Determinationof DimethylnifrosamuwandEvaluation of 8—314(DETA) In theAmes Nitrosoprolineby Differential PulseSoimonello/Mici’o.oare-PlateTest.Final Polarography.” Anal. Chem.47(11): 1857-Report.”1971 -(Submittedto Dow Chemical 1060.1975. -Coinpeny). -- - - (20)Fan T.Y,. )CrulL I.S..Rosa.R.D.. W0IL

(5) Ayanaba.A,. andAlexander.M. M.}L andFine. D.H. “Comprehensive‘l’ranaforinetionsOf Methylsniinesend - Analytical Procedureslot theDetermination

- FormationofaHaxardousProduct. - of Volatile andNon-volatile,PolarandNon-Dtinethylnitroeemine.a Carcinogenand polarN-NitroeoCompounds.”InMulagen. in Samplesof-TreatedSewageand -- -“EnvIronmentalAspectsofN-Nitroso

Lake Wrter.~J. Environ. Quit 385-I9.1974. - -‘. Compounds.”Walker.L.A., Grictiata,L,(6) Aynniibs. A,,Veretrasta,W., and - Caste~nird.M., and Lyle. RI..eds.IARC

- Alexandcr,H, “Pormatiànof , SCientificPublldatlonsNo.19.Lyon,FrancsDtm.tky1nlb’oumi~ie.a Carcinogenand ~.RC 1971Mutageu~isSoilsTreatedwIth Nitrogen - - (21) luenbarg,P.“AnalyticalMethodsforCompounds.”SoilScl~’Soc,Amer Plan. O~S1flh1isL~ “N-NZttVSO Compounth.’~ iqpu, Scanliri.LA., *n(Taiuenbeam.S.R,ida.

(7) Yordy) and Alexander lit “Formation ACSSymposiumSeries174. Washington.äfN-Nifrdtnditthanolamln.from - D~C.Amirecan Chemical Society,1981.

Diethanoinnilne-In LakeWater.-‘4 ~i,yage,~’ (22)Bollag, ).-M.. andBsrsbesz,W. “Effect‘J, Ent,ftctm, QusL10.26e-271~ - ofHeavyMetalson theDenftrificatlon

(8) Drutkre~?,H, Preussinano.3.. - Processin SoiL” J. Environ. Quit atlas-mi.Ivankovic.S., SchmahlD.. Aficham.J.. Slum. ~ -- -G.. MenneLH,D.,Muller. lit, Propoulos.p,, (21)Boilag.).-M..andHenninger.NP.tand Schneider.H. “Orgsnotrop~carcinogen. InfluenceofPesticideson Denftrlflc,?donin

Wirktingen hem65 vemuchiedenenN. Soil andwith anIsolatedBacterium. j.Ntti’oeoverbtndungenandBD.Ratten?(In Environ. Quit MS-lI 1975. -GernianlZ~Kribefóracb.09’.i03—201.ltd?. (~)~ S.. Watanabe,L. Honma,M.,

(9) Hunch, j., Schmaltz.L, andHoffman1). ~ Honda. S “TheEffectofPesticideson“Effects of N NitrosothethanolammaandLi ‘the Denitriftcatlon In Paddy SOIL” Soil Sci.DiethanolhydrazumeId Syrian Golden - 4lItt Nutrition 1~15-23.1954.Hamsters.’CancerLetters4.55-80.1975. - Bollag,1.-lit. Orcutt, M.L andBollag.

(10) Pretiumana, L Nab.. M.. liabs,H.. - ~. Denitriflcatlonby IsolatedSoil Bacteriad Schin h1 ~ “C~In - - - f- ~ - - underVenousEnvironmentalConditions.

Nitroaodiethanohamine in Rats at Five - - -‘ -- - -DifferentDoseLevels.” ~ ne..4na107- (25)NTP. NationalTozmilo~rPro~ani.

~ 1962. - - - - “Sslmonella-T.stthgResultson- - 11 Boil - - and-ti S. “N’ Dlethyleneu’iamine. 1964. -

andNitrous.OxideA~cumuld” - an re5caDenitriflcationin thePresenceofPeaticadi - “MutagenicityScreeningofIndustrial

£ ~ - i.~ ~ - ClmemicalaSevenAliphatic AminesandOnellVa i~5. ..y~ Vi~Ofl. tO - £ d 1’ eA - 1. 0 1 iI 1

39-845-8491980. - i a as in a one a, crosom- - . ,. Assay.”MutationRe..53.198-196.1975.~12) oeuacn,i~i.n.,aria imeuje. J.M. ,..m,tlc .. - -

~ i .~ ~ - - ~. -- j.~jmCeOitoxic nuustancas.pane on or ono i -Nitric Oxide.andNitrous Oxide durins - -V ~ITt - A - - 8 01. ArlIngton. VA. NatlcnalTechmcsl

- - - -- C en ni ca on. .-.~, viron, - ~ f - - 0.4 ~. ~- MicrobioL 42:1074—10641961 - ci -- o.

- - - - -(13)Stanier.R.Y., Melberg,L.A., and - - -2129 ~u$uaL1982’Ca~-‘ I ‘ M.L.. el-al. ‘~Unzcbsdu1edDNA Synthesis

- EngiewoodCliffs. New Jersey:Prentice-Hall. Tests:a R~poflof theU.S.Environmental- Inc., PP~~ .. ProtectionAgencyGene-TaxProgram.”

~ ~ - - °n - - Mutation Res.1~383-410.1963.BlodegridabditycfPidyammass.”“~ (30) Trubko~E.I.~andTeplyakova,L.V “ADel. 1t310-311.lV7a (TrsñaiatedfoeEPA by Studyof Di.thylanetriamlnsfor Purposesof

Uterature Research(pony). - Hygienic Standardizationin Bodiesof --(15) ‘t’?~tIY,J~R~ ~ ~ - - Water.” Gig, Smut. 37:103-104.1172..

“Microbial Metabolismof N- - rr~ I ~ hv lãM.a~stuyuResearchNltrosodlethanolamltlsin LakeWaterand’ for EPA). -Sewage.”AppL Environ.MicrobiaL 39:559— - - - - --563.19~ - - - -.. - IlL OtherRegulatoryRaquk-~~~’-ET1 w361 235 m411 235 lSBT

- -- (16)-Tate.RL,-IIL.andAlexand.r.M.- .“Stability ofNltroeemmeaIn SamplesofLake ‘~‘ ~ “er

— Water Soil,andSewage.”J Nat. CancerInst. Under ExecutiveOrderi~, ~A54.327-330.1975. must judgewhetheraregulationIs

- - (1.7) Popp.Kit BASF. AG. Ludwiphafemm. “major” andthereforesubject-tothe- - - .-- Germany.latter toLL laCks.~ re~uirementof a to In ct --~ : - - -- ChemicaisAssesamentDivisi~Office of - .

PesticidesandToxicSubstances.U.S. s,ua.ysls.~ ~u~u0U torEnvironmentalProtection1~gency chemlcal4ubatanceIs not maor becauseWashington,DC W480.1984(March 13) It doesnot meetany of theaRenaset

(18) Dahmen.E.AJ.LP.VaderD.. and Van forth in marion 1(b)of theOrder First( dirLaarss.j.D lime Polarographic theactualannualcostof thetesting

prescribedfor DETA is leesthan8128.243overthe testingandreimbursement period.Second,becausethecoatof the required testing will bedistributed overelarge productionvolume, the rulewill have only veryminor-effects(lessthan 0.31percentayear) on producers’costsorusers’pricesfor this chemicaLFinally, taking intoaccountthenatureof the marketfor thissubstance,thelevel-ofcostsinvolved,andthe expectednature of the -mechanismsfor sharing thecostsof therequired testing.EPAconcludesthatthere will be nosignificantadverseeconomiceffectsof any type u a.resultof this rule. ‘ - -

Thia regulationwassubmittedto theOfficeofManagementandBudget(0MB) for’ reviewasrequired by - -ExecutiveOrder12291.Any commentsfrom 0MB to EPA, andanyEPA - -responseto th~necrunments.are - -includedin the public recordfor thisnil.. - - -B. Re3uIotoryFlexibihtyAct

Under theRegulatory Flexibility Act(15IJ.S.C,601.-Pub.L. 96-354,September-19.1980),EPA is certifyingthatthis teat -rule, if promulgated,will not havea- -

- signtflcantimpact ariasubstantialnujnberof small businessesfor thefollowing reasons

1.-Basedon the’EconomicImpact -Analysis preparedfor this rule, thereIs

-only. one small-manufacturerof DETA- that-~mm~iufacturuylessthan00~

-- percentof the-estimatedannualdomesticproductionof BETA. Althoughno figur,s areavailableto indicatewhetheror not therearesmallbusinesseswhich import DETA. the -total amountofBETA imported Isestimatedto represent lessthanIpercentof theestimateddomesticproduction of BETA.Thus,theestimatednumberof small -manufacturers(Includingimporters)affectedby thi. rule will be quite small.

2. Small manufacturer,andsmallprocessorsarenot expectedto performtestingthemselves,or to participate intheorganizationof the testingeffort.

3. Small manufacturersandsmallprocessorswill-experienceonlyminorcosts,If any,In securingexemptionfrom

- testingrequirement.. -- 4 Small manufacturers and small - -

processor,areunlikelytobeaffectedby- -- reimbursementrequirements.

- CPaperworkROductionAct-TheInformationcollection

requirementscontainedin this rulehavebeenapprovedby the Office ofMaimagemintandBudget(CluE)under -

- - - the prOvisionsofthePapa~rk - -

- Fsdor.~R~atss/ Vol. 50. I’~.100 j Tbm’edsy.May 23, I8~5f Ru)es andge~u1etfnrm,-

ReductionMt cL18~.4* U.S.C.35~1.s/ seq.sad havehensa5~eadmrmbe2~-~-

List dReb~ecteIn 48CP~Pt~IITesting.Envir~antsLpsof~~n.

HazardousmateriaL~en~-

J.A.M~.~ ~.dTozic54~—- - -

PW 78*4MRN~IDIThesafbrs4~~RPart7~is~“~‘ed

asfollower1.Theauthuiltycft’ttlan farP1st748

continuestoreadasfollows:AutSorlty 15 U.S.C.~O3.2811.~2L2. By addingI 7~35~5~ senden

follosen

5788.1879

Dlsthyls.strtem8is~ETA)(a) Identificationofchemicaltest

substance.(1) DIs*yIene~tamien(CMNo.1Th-40-O.ale