Thursday May 23~1~S5

Part~ Environmental. Protection Agency 40 CFR Part 79S Dlsthyl.ne1flamine identification ~ Specific chemical Substance and MkU~e testhig Requirements; Final Rule Dl.thyfert.trlamlne; Proposed Teat Rule; — Rule

JI~ 21398 Federal Register f Vol. 50. No. 100 1 Thursday, May 23. 1985 / Rules and Regulations ENVIRONMENTAL PROTECTION chemical fate testing (underaerobic relevant to assessing the risks to health AGENCY conditions only) for DETA. and the environment posed by exposure 1. Introduction to particular chemical substances or 40 CER Pal 799 This notice is part of the overall mixtures. implementation of section 4 of the Toxic Under section 4(a)(1) of TSCA. EPA (OPTS-420129 19*4-FRI. 2815-Sal Substances Control Act(TSCA, Pub. L. must require testing of a chemical 94.-469, 90 Stat 2003 etseq.. 15 US.C. substance to develop health or Idantificatlon of Specific Chsnilcal environmentaldata if the Administrator Substance and Mixture Testing 2801 et seq.) whichcontains authority R.qulrem.ntLD4.thylen.thamln. for EPA to require development of data finds that: AOSNCY Environmental Protection (A)(i) th. manufacture, distribution in commerce, proc. Agency (EPA). eonng, use, or disposal of a chemical substanceor mixture, or that .*CT*Osc Final rule. any combination of such activities, may present an unreasonable riakof injuryto health or the environment, (n) there are insu~cientdata and.aqerience upon which the ai~anv:This rule establishes testing effects of such manufacture, distribution in commerce, processing, requirements under section 4(a) of the ma,or disposal.of such substance or iI~iThweor of any combine- Toxic Substances Control Act(TSCA) tion of such activities on health or thf.Invironment can reason- for manufacturers and processorsof ably be determined or predicted, and diethylenetriamine (DETA~CAS No. (di) testing of such substance or mixture with respect to such 111—40-0) consistIng of (1) oral sSscts is necessary to develop such data; or subchronic (90-day)toxicity in at least (B)(i) a chemical substance or mixture is or will be produced one mammalian species, (2) dermal insubstantial quantities, and (I) it eaten or may reasonably be absorption in the same mammalian anticipated to enter the environment in substantial quantities or species used for thesubchronic testing. (II) therein or may be significant or substantial human exposure (3) chemical fate under aerobic Scsuch substanceor mixture, conditions, and (4) mutagenicity (ii) there are insufficient data and experience upon which the (includingtests for both gene mutations effects of the manufacture, distribution.in commerce, processing, use,or disposti of such substaflee or mixture or of any eombina- and chromoaomalaberrations). This tion of such activities on health or the environment can reason- Phase 1 final testrule constitutesEPA’s ably be determined or predicted, and final decision concerning the testing (iii) testing of such substanesor mixture with respect to such needs for DETA as recommended by the effects is necessary to develop such data. C InteragencyTesting Committee for all effects except carcinogenicity. Elsewhere in this issue of the Federal For a more complete understanding of above. DETA reacts withCO, inthe ai Register. EPA is proposing under section the statutory section 4 findings, the to form carbamates which precipitate 4(a) of TSCA that DETA be tested in reader is directedto the Agency’s first ‘from solution. For this reason, DETA i~ chroniconcogenictty bioassays, If this proposed testrule package manufactured In an essentially closed substance exhibits positive results in.. (chloroaietb.ne and chlorinated system and is transported under a certain of the mutagenzcfty tests beuzenes. published July 18.1960(45 FR nitrogen atmosphere. Due to DETA’s required In this final rule. 48510)1 and to the second package reactivity with airand to the conditior oaiu In accordance with 40 CFR 23.5 [dishloroinethane, nitrobenzene, and used during its processing, the Agency (50FR 7V1). this rule shall be i.1,1trfchloroethane. published Jima 5, concludes that it is unlikely that promulgated for purposes of judicial 1981 (48FR 30300)) for in-depth significant emissions of DETA to the review at 100 p.m. eastern daylight time discussions of the general issues atmosphere will occur. On the other on June 6, 1985. This rule shall become applicable to this action. hand. EPA believes that significant effective on july 8.1985. IL 3ad~powid releases of DETA to waterwill occur ~esmana~su~noecow~*cY during manufacturing and processing Edward A: Klein.Directar. TSCA A.Pt’Ofile operations. EPA believes that Assistance OfficeCrS-799). Office of DETA. CASNo 111-40-n. Is s~ occupational exposure to DETA ToxicSubstances. Ra. E-543, 401 M St.. r alkaline. hygroscopic~viscous liquid. (primarily by thedermal route) occurs SW.. Washington. D.C 20480. Toll FreeV~ The estimated annual production of during manufacturing storage. transp (800-424-9065). In Washington. D.C. DETA in 1982 ranged from 28 to 3~. processing, and clean-up activities. a, (554-1404). Outside the USA: (Operator. million pounds. The primary uses of • that themost likely source of consum 202-554—1404). DETA are for the production of paper exposure to DETA consists of dennal su~.suvrrasvNWOSMAnOse In the wet-strength resins, epoxy-curing contactwith epoxy-resin products FederalRegister ofApril 29,1982(47 FR agents. cheIati~gagents, lubricatingoil containing the substance as a curing 18386), EPA issued a proposedrule and fuel additives, surfactants. and agent. under section 4(a) of TSCA to require corrosionInhibitors. DETA also hata L JTC Recommendations testing of DETA for a variety of health minor use as a decontaminant for effects and for chemical fate under both military chemical agents. As discussed The Interagency Testing Committe aerobic and anaerobic conditions. in detail in the Agency’s - (lTd. organized undersection 4(e) o~ Today, under section 4(a) of TSCA. EPA Diethylenetriamine Support Documeni. . the Toxic Substances Control Act I Is promulgating a final Phase Itest rule which is available from the TSCA tTSCA),Included DETA in its Eighth requiring health effectstesting and Assistance Office at the addressgiven . Report to-the Administrator of EPA / V FedaaaL Register / VoL 50, No. 100 1 Tht~rsday,May 23. 1985 / Ruins and Regulations 21~

April 2& i9~1.p~SdE’thSF.dmeL resulting from DETArelease to the not the Agency’s choice of the ma!route Register of May 23.1961 (46FR23139). envirr~nt.and ch~i.&lLate testing of exposure for therequsud scbchroni~ The ITC designated DETAa. apriority (under both aerobic and anaerobic studies appropriate? chemical and re rw,i.~sndadthat it be conditions) is rV’.c4~ tcdevelop such d. Aithounji theAgency is spectfyiag testedfor healtheffects, to data. the oral route of exposure for- the chroniceffects. repro ~ I&~-E~t~. and 3. Differences from 17’C.g requi,ed iubchronic [90.day) testing of teratogenLciLy~The LTd 0based itt Recommandaticna. lithe proposed rule DETA. the Agency is primarily desi~*tionof13E~Aon the substances for DZTA.theEPA also prc~~the concerned about dermal expos~’.vto known bioiogical efTacts, the reported reasons why theAgsocyspro~e.ed this chemical: would it. therefore.not be • produclianin exams of10 milhioc testing.ieq~akemeninfor DETA differed necessary to require the performance ci pounds per year. and theNational from the TICsosmameedabenahe the a dermalab.rpttonstmiy of DETA to Occupational HazardSurvey (Ref. 1~ ______U follcwss provide date aesded in e’uabests the estimates that 63,000workers are. o.L~Acyproposaksuhehreinc risks posed by dermal expoasond potentially expoosdtc DETA. testing rides than -iletini, ~eeic s. A1i1 theTIC ie~~~-~’ stodion became theAgeeq lmMe,sa hill-~eti.~ hsla8 ofIJETA. C Pru!~ thes ~p—~y ~ 60.diy ~ would nil ~k nd~ (96-day) tanicity • EPA Issued aproposedrulepubLhed withse~~mi%m testing in”-~g~1~ cespr~nAve In the Fss~afXApr~2L1I6Z mayb.ssodan... ______histopathokgisel ———-—tics ofbody (47FR18396) in ~ lathe instfr~ Iifeti~estoc~at~aM tissues, be .d~- b~.ce~LETA- recammendatlcrssbytheTICon~rA. effectsother th~~riri~p ~ related effectsthat would be ob...... ,I 1. TestR U1c~JaE~The pru,u..ed certain other age-relatedeffects.The In fu1J-hfe~e~ 1rut1x~.~ he rule requiresthat DE’TAbe tastedfor~ available data providedno sound bans thosee~.sinre.,Jii..~long latency a. Subchroaic (g0day} health effects for supensise .CDETAs’ abilty to case. perkida? In at least two man~han.p... oncs~~orags-islatedeffects. b.Mutagen~cltyf~enematationand b. EPA ~Enot prepme testirqfar D. NewDev’~’~ Fo& cy ___ reprsdactiw sad ~ effects. Proposed Rule becasae, hr ~Agmay”, ~ di. Theproposed rule farDETA ~‘PR and anaerobicmaditiessj. available data çaMIr.~limited)did not The EPA based Its noposed testing 1838& Ap,Rm 7961~indicotedth.t.lf suggest .poinntia~ha these efisota. interestedpertlea ,~uoded—• reqm~me!~. on dieauthorityof section The analysis and find~onwhich the 4{a)(1)(AJ ofISCA. opporr~eyfaroralca~e~co die above determinations were based ire proposed refe. then theAgeniy~M 2. Findingn TheAgeecy fosodthat the preen~trdfri the~eby1anetit~ne a meeting en thisr~ to manufec~e.~ ass. and Support Do.~nt. whichIs aenthble WaahIngtee~DC.. en July 13.1962. Since disposal ofDETA may presentan from the OfIn,olTcxfcSithstancus’ ( unreasonable risk of injury to no requestswere received by the EPA health, dns in ~ and iwsagenfa ‘rSCA A~efatancaOffionThe rrc, for thepresentationoforal comments on recommendati~and EPA’S ~ this rule, an peb cmeetfugwea heft A effects. for thefollowing resson~ testing requirementsma a. There ma e~istlng~te winch meeting betw~en ,.-..ntatlves from Indicatea poinutial ~n health below~ the Union Carbide Corporationend hazardh~ DETA Mth re.~.atinthese members ofth sdenti~cstaffof the ~c ~n Office of Toxic Substances ofEPA effects. • o-_ -~ .~ — b. EPA believes ~ era -~-g-• me~ held an SeptsmberlO 198Z in dIscus, mutagenicitystudies ofDETA (1ef 3) exposed in DfFA In the kpl~.in ‘ a a using cenaiwsa products. ~ , performed for that armiofactusenwhich of release oi DETA into 5. are discussed in datailIn Unit 1IL F. : I 1. kidss~yComments.The Dow environ~- •••-___t a C. The~ecp also fe ~t ~ ~ a Chemical Company aridthe Union are ~enr~t data in l~2 the Carbide Corporation submitted the subchrsoie d ~qc clTi~~f following additionalrelevants~n in DETA. ~ inedugoL~Ain~-----—, 4. bsassfrr C~~a izrPr~,po..d the Agencytogether with these SrmV ~ d_~ Rule, In th.proposed t.sti~ for D~TA.. comments, dated June ~ on the in andi~a.~A ~ the the~ecy raised thefoliewingm.~ce proposed test rule for DETA ~-- —i. ass. Issues fokc~~w.~i~ a. Structure!and ~ologlcal Activity disposalof DETA p—& eShedintydeonodiar Relationships BetweenEthylenedlamine unrea~l* i~ to ~ ~bk ethyIensia~~~ at and Diethyfenetriamine(included to on~ ~4Lc~ s&L~ta~by theN- ethyIenedi~esod preliminaryabsorption. distribution. nlfroumine derivative of DETA. for the fri.thylenstetsamine} beused sea metabolism.4 and pharmacokinetics following surrogate faaDETAtonicity? studies of ‘ C-radiolabelled DETA and d. ManyN.eitios~~hew been b. Whet protosni. s~eI~be used fez ethylenediamine following oral or shown to besereinogenlc.1~-ar. the chemical faintes~ of D~E&~n.d endotracheal admini.tratfon to male existing~e whish indicatoa &t quantifying de extentof de rata}. theoretical potential for theconviwian biological (or chemical)lyaaslormaklgn b. A~dascrIpthm ofstudies of DETA in .N.ait,osmaien in the of DZTA to anIJ.introsamine derivative perfomed.by Dow Ch,,nit’i~1 Company to envfronmailand thet ~ona map be ofDETA by sir zgsaisxan pres~In determineifs Nm’trma~ederivative exposed tothi. AT-ultr.samine as a water. sew~.and sail? of DETA would form in an equeaor in reseM of thesuleas. of DETAindi. c. Becama of dithouMias InvolvedIn vitro chernicaL~de1system. envfrenmenL quantitming thedeseQf~A thet 2.SectionS(d~lS~efucm2~ I n Th. date.., ~ien&to predict animals oofd ~ve.in ~aA 29&3. In. a.TSCA “ ~ ~ the ~ of subchreele~96.day).t~,, ~f~gI’~is- on DETArscaLvadbydeAgan~on. 21400 Federal Register / VoL 50, No; 100 /. Thursdtiyv May 23, 1985 / Rules and Regulations

June 24. 1963, UnIon Carbide DErA.~both of the submitted studies properties, these two substances are Corporation included the following contain only verylimited histological likely to be handled similarly In - ( additionalrelevant reportm data. biological systems. a. Pharmacoklnetlcs and Metabolism The Agency barreviewed the data on Dow and Union Carbide submitted ti of Diethylenetriaminein theRat.(A DETA contained inItems3.a. and 3.b., results obtained from studies using ‘~C complete reportof preliminary studies In and ha, used these data in reaching radiolabelled DETA and WA and Iteml.a.). conclusions regarding the ability of aimed at determining the excretion b. Dlethylenetriamlne-Comnrerclal DETA to Induce specific locus mutations pattern, the tissue distribution, and the DermalCarcinogenesis Study In Male (at the hypoxanthine-guanin. blood-level pharmacokinetics of the C3HIHeJ Mice. phosphoribosyl-transferese locus) in radioactivity observed following the C. Diethylenetrmamine-HIgh Purlt~ Chinese hamsterovary usia (see oral or sodotracheal administrationof DermalCarclnogenesis Study in.MeIb discussioninUnitIILF.). In addition, the these rsdlolabellèd compounds to C3H/HeJ Mmon - Agencyhas reviewed the information on Fischer 344 rats.In addition, the urinar 3.Aai5ctiontd)Subinisslonz DETA contained in lion tb., and has radioactivity obtained following the March ~ bra~ - concludedthat these date donot negate edmI,~L~trationof rsdlolabelled DETA submission us DETA received by ~. the nicesnity forch.miea-I fats tasting ci and WA torats was characterized by Agency en March 22. 19S4. Union DETA todetermine if saNos~~fr~ 1on.evrl~engechromatographic methni CarbideCorporation Included the derivatlv. of this substance could be Dow and Union Carbide interpretthe following reportand associated formed under environmental conditions results ofthese studiesas Indicating ti document (see discussionIn Unit WI.). a. Summary of Exploratory Testsat DETA and WA have the urns genera ilL Public Comm~t’ pattern of disposition In rats. and that BRRC (Bushy Run Research Cents,) DETA and its metabolites are more with the CHO/HGPRT (Chinesehamster The comments receivedby the rapidly eliminated from rats than EllA ovary cells/hypoxanthine-guanine Agency in response to the proposedtest • and Its inetabolitu. Thes. firms phosphoribosyltransferase locus)Test rule for DETA were from theaffected System. Industyand trade associationsources. submitted to the Agency the results of both a 7-dayrange-finding feeding its b. A memorandum to Mr. Di.. The majorissues identified during the Heywood..UnionCarbide Corporation. comment period are discussed below. in rats of the dthydrochloridsof WA.. from Dr. Ronald S. SleuinskL Bushy Run and the results of a 3-monthsubacute Research Center. desathiag how the A. Appropriateness ofUsfr,gAnalogue feeding study of WA dihydrochhatds results of the above study relate to Data to Assess DETA.’, Toxicity rats,and believ, that an extensive. various EPA guidelines and U.S. EPA One of themajor issues for which the toxicity testing program for DETA Gene-Tox Program reports on this assay Agency requested public comment in the should be delayed until th. results of system. proposed test rulefor DETA Is the further toxicological testing.on WA. Items l.a. and2.a. have beenjudged appropriateness of using available become available. ( by the Agency as insufficient evidence toxicological Informationon The Agency mustdisagree withthe that toxicity data for ethylenedlamine ethylenedIa~~~in~(WA) and major manufacturers of DETA that tb~ can substitute for the required toxicity thethylenetetramine (1TrA). proposed proposed toxicity testing of. thin testing of DETA (see discussion in Unit structuralanalogues of DETA... a substance should await further lILA.). substitute for testdata on DETA itself to Information regarding the toxic effect 4. EPA Review ofSubmittedBioassay assess the potential toxicological of EDA. While DETA and WA are Data. The Agency has carefully hazards posed by DETA. The two major similarin that they both possess two reviewed the two dermal manufacturers of DETA in the United primary amino groups.they are vastli carcinogeulcity bioauayaof DETA States, theDow Chemical Company different withrespect tothe factthat (DETA-Commerclaland DETA-High (Dow) and theUnion Carbide DETA also possesses a secondary Purity) submittedby the Union Carbide Corporation (Union Carbide), submitted amino group, while WA does ant., Corporation (Items 2.b, and 2.c.), to th. onlycomments addressing this Issue. Substances having secondaryamine. determine If these studie. were Thesemanufacturers believethat WA groups are well known to be muchan performed in a mannerwhichwould I.a close s~ucturalanalogue of DETA. susceptible to stable N.nitrosamlne negatethe need for oral subchronlc (90. stating that DETA may be regarded as formation than substances possesath~ day)testing of DETA. The Agency has the dirnerof EDA. and that both only primaryamino groups.Other. u concludedthat these studies.are substancescontain two primary amino yet unknown, differences with respss inadequate to negatethe need for oral groups, while DETA (but not WA)also the production of toxicinetabolites n subchronictasting for thefollowing / contains a secondary amino group. Dow also existfor substances containing reaions and Union Carbidebelieve that the secondaryamino groups as opposed a.In bothof the submitted studies, chemical behaviorof both DETA and those containing only primary ammo only male micewere used; thus, sex. EDA will be dominated by the presence groups. relateddifference, In response to DETA of the two terminal primaryamino In addition, studies submitted by administration could not be groups In the substances. Furthermore. these manufacturers do not, In fact. investigated. these firms point out that the physical demonstrate that the metabohtas •b~In both studies, onlya single dismal properties which Influence behaviorIn • produced by rats from DETA and El~ dosage level of DETA was employed; biological systems are also similar for are the same, since the radioactive thus, no dose-response relationships the two substance.: both ire completely metabolites appearing In the urine withrespect to DETA.related effects soluble Inweter. both form basic • following the administration of the ti could be Investigated. aqueous solutions, both ayerelatively radlolabelledsubstances hav, been c. The Agency ii requiring nonvolatile, and both are low with compared only by ion-exchange colt1 comprehensive hlstopathologlcal respect to molecula, weight These chromatographic techniques which, (- examination of tissues in the required manufadw’~ribelieve that, because of -. themselves,do not allowstructure! - oral .ubchronic (90.dayjtesting of these similarchemical and physical Identifications. Even thiscomparisot

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

flawed by the fact that different elution produced from WA by rats. In addition. EPA by Union Carbide indicate that, systems were employed in the cohmin even if data were available to DETA is absorbed following oral chromatography of the radioactivity demonstratethat identical metabolite. administration, led the Agencyto ( foundin the urine of ratstreated with were produced from both DE7A and conclude that theoral route of radiolabelled DETA or WA, making WA by rat., analogue data available administration should be required for comparisons of radioactivemetabolites for WA cannot currently be used to the subchronic testing ofDE’FA. Only by their column elutlon volumes arrive at expectedpotencies of DEFA Dow and Union Carbide commentedon impossible. In addltion..the column for thehealth effects of concern. this issue. These manufacturers agreed chromatographic profilefor the Therefore, based on the above with the Agency that oral studiesof radioactivity foundin the urine of rat. considerations, the Agency has DETA would allow the adequate treated with radiolabelledDETA concludedthat it would be evaluation of thesystemic toxicity of contains at least sevenradioactfve~peà Inap~~pil.teto utilize the toxicity data this substance withoa~thedifficulties of fractions, while the.correepoodi~ available for WA (or other proposed determining the effective doses received chromatographic profile’for the--~. structural analogue, of DETA. such as by treated animals which would arise In radioactivity found in the urine ofrats -• TETAJ to assess the potential dermal studies. In addition, these • treated wtth radiolabelled~A r~mtaiiig toxicologicaihazards posed by.DET& - manufacturers pointed out that the - andthat testing of.DETA Itself Is. • only threeradioactive peakfractions. - knownskin Irrttancy and sensitization This fact may well indicatethat DETA necessar~. - potentials of DETA would likely lead to is, in fact. metebolizeddifferently than B. Appropriateness of Sabchrvnic stressfulconditions In pniTn.l$ receiving WA by: therat, In any case, thedata Rather than Chronic Testing ofDETA DETA by the dermalroute, making the from these studies do not Indicate that evaluation ofthe systemic toxicity the metabolites produced from DETA Mother Issue for whichthe Agency observed in such studies diffiCult These following oral or endotracheal requested commentsin the proposed difficultieswould not aria. In oral administratioti to Fischer 344 rats.sie test rule for DETAin the feeding studies. Therefore, theAgency I. appropriateness of a 90-daysubchronlc - the same as those produced under the requiringoral 90-daysubchronic soximty - same conditions from FDA~.Evenifdata test duration ratherthan afull-lifetime chronictest duration for theproposed testing In the final Phase Itest rulefor were available to demonstrate that the DETA. • metabolites derwed from WA were toxicity testing of DETA. Thejnsjoy identical to those derivedfrom DETA. manufacturers-of DETA In theUnited. D. Necessity ofa DermalAbm,pcà,n States, Dow and Union Carbide. Study ofDATA theAgency could not, accept •, , - submitted the only comments on this • toxicologicaldata on WA as a . - - issue. Theposition ofthese Another Issue. which the Agency - substitute for the testing of DETA ~tielf. raised for comment in the proposed test because thereIs currently noaccepted manufacturers Is that an oral 90-dày subchronic study of DETA will suffice to rule.for DETA was the possible methodology for determining the necessity of requiring adermal potenaes expected forDE’tA for the provide adequate information on all systemic toxic effects whichwould be absorption study of DETA. sincethe health effectsof concern from analogue Agencyis primarily concernedabout • data available for WA or other observed for thesubstance In an oral full-lifetime chronicstudy of this potential hazards posedby this potential analogues, substance, with theexception of substance due to exposuresvi. the On the basis of radioactivity alone carcinogenic effects. The Agency also dermal mute. Only Dow and Union (saying nothing about thechemical believes that, In general, a properly Carbide commented on this Issue, and structures actually containing the conducted90-day study, Including these manufacturer, believe that such a radioactivity), the metabolism studies in comprehensive histopathology. can be dermal absorption study would, Indeed, the rat submitted by Union Carbide and used asa surrogate for the full-lifetime be necessary. in addition. these Dow indicate that both DETA andWA chronic study with respect to the manufacturers submitted a protocol to arereadily absorbed via the oral and detection of chemical-related health theAgency for astudy aimedat endotrachea! routes, distributed effects, except for those requiringlong determining thedegreeof dermal throughout thebody, and exoreted latency periods, such as carcmnogemuity. absorption of DETA In rats from data- primarilyin the urine and feces. These Therefore, theAgencyIs requiring obtained in disposition studies of the data also indicate that DETA and its subchronic (90-day) testing in thefinal substance, using both Intravenous and aretabolites are eliminated from the Phase I test rule for DETA. In lieu of full. dermal routes of exposure, body at a faster rate than ED&andIts lifetime chronic testing. - Sincehumans are expected to be • metabolites,and. In addition, that DETA exposed to DETA primarily by the and/or its thetabolltà~ar.retained to a C A~bpropriotenessofthe OralRoutefor dernial route, the Agencyconcludes that • lesser degree than-WAmd/Or Its Subchronic Testing ofDETA a dermal absorption study of DETA Is. metabolites In bodyI4.~.& • In the proposedtestrule for DETA. In fact, necessary in order to assess the In sathe-marderlying theAgency also requested comments on hazard, posed by DETA by this route of mechaniiin, responsible for thetoxic EPA’s selection of the oral route of exposure, andIs, therefore, requiring effects noted by Pujino{ReL.2).. exposure as the route of choice for the such testing in the final Phase Itest rule primarily In the livers,,kidneys, and required 90-day subchronictoxicity for DETA in the same rnamm~flan lungs of Wistar rats receiving chronic testing of DETA. Although the Agency species selected for the required oral - treatmentwith DETA via the dermal or believes that exposures to DETA will subchronic (90-day) testing.. -‘ subcutaneous mutes. are as yet occurprimarily by thedermalroute, the unknown. Tho Agency has no evidence -difficultiesassocm,ted with determining £ Protocols forRequired ChemicalFats • to indicate that the metabolites theactual doses of the test substance Studies ofDETA - produced from DETA by.rste(which received by theanimals In studies Thefinal Issue for which-theAgency may be.responsible for some or all of the utilizing this route of administration. prquested comments In its proposed test observed toxic effects’of this substance) together with the fact that preliminary rule for DETA involved which protecols are the same metabolitesas those pharniacoklnetlcs data submitted to should be used forthe aheetica}fate. 214$2 Federal Regiater / Vol. 50, No. 100 / Ththday May 23, 1985 / Rules and RegulatIons

studies the Agency is requiring for juagneticresonance spectrocietry. Dow nitrosamine derivativeof DETA does DETA. Since the miciobielformation of - concluded from this preliminary not form or. if formed, rapidly degrades. N-nitrosarnitmes from secondar~amines experiment that almost all of the original These firms also believe that thescope present in water. sewage,. and soAlhu DETA had disappeared from the of work involved in answering the beenreported by several lavestigeters reaction solution, but that no- question in a quantitative fashion is too (Ref.. 5 thruugh71~.tbmAgency is characteristicUV absorptionband at great to be within the proper scope of a concerned that an N.ni~o.a1.1l1t. 300-400nrncouldbe detected to TSCA section 4 test rule. derivative of DETA aa~be firmed In indicate the presenc. of anifrosainlne. With respect to this issue. CMA these eiwiiooments~upecially in Dow concludedthat it is unlikely that commentsthat EPA should avoid sewage treatment facilities), may be theN-nitros~reof DETA mm be requiringtesting under TSCA section 4 transported,4 to4waterenirces used for - produced underaerobic conditicos. or. If in cases lnwhich thetesting the pco~ a~t nnof king.w,ten..~, pmduced. theNoifrosamnlneaI~lA methodelo~fsnot sufficientlywell suavivedrinhing-water treatment mustdomanpom rapidly. developed. Specifically, OdA believes procsds.aud.ihus. ~a potential- With respect to the anaerobic that the chemical fate testing requiredIn carcinogemchawd to the4 general1 - biotransformation ofDETA to an N- the proposedrule (aimedat determining public by its presenea n’ ’~n ’ngwaton njtrmmmine derivative, UnionCarbide - lien N-aitmuminels produced hoes A clom sthic*maral enelegeeof theN- andDowb.lievethistobshlgbly - DETA by’mlcverganismaimdsr aerobic nhtrosamine derivathsdDEl’A,N. unlikely, sinceDETA (which is very orarmarobic oondltions)1. -. fliodith~n~~LsawL1~ELA),e is a watersoluble)wouldnotbe expected to-~ Inappropriate, simc, to DlL’~s - - - kno~wt~ cerc1msen3 (Rd.. a sorb to soils andserlimAnil having... knowle~,there I. ao~rentstenderd through-ID). In sdcbboe, 4Yerdyand anaerobic environments; additionally. methodology foi’pcf.~.gthese tests. Alexande~Rd~.Jhaea —~tr.ied these firms believe (citingRefs. 12 CMA belleveehtks* theresults from that NE~LAwa.foxmed from through13) that any nitrites present In these tests we unlikelytobe sufficiently dletKanoIamine in lakywatar end-In such environments would be reliable for u.s-in EPA’s sewage, andthat aileaaLsoasoI this- metabolized by mioroorgamsins to’ dedEoelongrthv~G~rw.suth~tes~ ND~.Aprcelucboa wasprobably due to nitrogen gs~thus removing a required fellsoutside the anop.of ‘tvhat can be- the actionof micmoorganisms or some reactant for nitrosamine formation. requlrod-under-TSCA. other heat-labile factor. These data also Union Carbide and Dow believrthat Certzln of thecomments de.mibed-’ Indicate that some ofthe ND~.A chemical fate testing for the fôm ation of above Indicate-uncertainty or ~~diieiOzr’ productionwas due to purelychemical an N-nifrosamlne derivativ, of DEM by concerning theAgency’. rat1onali-~ (nonbioIo~cal)react~ Whether the mi~oorganismsunder aerobic theproposed chemicalfate testW at N-uAliosamin.derivatives of conditions is afar reachingresearch DETA. as de,albed in thepropesed test diethanol.~vth~~ci DETAare formed by effort which Is notwithin the scope of., rule~ titlesubstance. In-actuality, the purelychemical or biological meanshas. TSCA sectIon4 testride. Thea, firms Agency is concerned-about thetotal’ no bearing on thetonichazards pøssd. claim that no standard methodology for transformation. whetherb*ologfcal’or’- ( by theN.aitro.amine derivatives determining low concentrations ofN- pisely chemical in esters, ofDETA themselves.Therefore,theAgency nitrosamlnes in water currently exist.. present In water, sewage. or’soils teen proposedchernicalfat. testhngofDETA. and that the synthesis and N-nltrosaminederivativeof-the under both aerobic and anaerobic characterization of the N-nifrosaihlne substance, whichthe Agency viewsas-a conditions, to quantifythe amount ofthe derivative ofDETA would be required. potential cardumogen sad whichmay - -. potentially carcinogenicN-oafrosaziine prior to the development of an enter thedrinldng water ~ derivative of DETA whichmight form analyticalmethod for the detection of With this clarification regarding the from DETAIn water, wage. andsoil. theN-altzosamina derivativ, of DETA In Ageucy~sconcerns about thechemical- TheAgencyinvited commentson water at thai ugh leveLIn addition, fate testing of DETA, EPA disagree. - experimentalprotocols whichm~tbe Dow and Uman Carbide believe that th. with both Dow andUnion CarbIde~as used for this purpose. andreferred workof Yordy andAlexander(ReL 7)1. well as CMA, that appropribte - interestedparties to themetbodol~es Insufficient as a model to use for the methodology doe. not exist, or reported in thestadies of Yordy and biotransformation study. sin~ethe be easily modified. for the successful Alexander(RL 7). method does notdistinguish between completion of this testing rm~uii~ent - Comments Were receivedon this leg.,- biological and chemical production ofN- In addition, the Agencydisagrees with from Dow, Union Carbide, endthe nltrosamines, and that anew. - the manufacturer, that the-r.sults of the Chemical Manuftcterer~-Ass.datfen methodology would have to be chemical studies perfonue~Iby Dow. (Q4A). Dow and U~ Carbidebehave developed for studying the - aimedat determining IfanN- - that the proposed ~ tasting. biotransformation of DETA to anN- nitrosamine derivative ofDETA could- of DETA aimed at ds~~g if aN. “nhtrosamine derivativeandar real be formedfrom DETA in aqueous nifrosamine will be produced ~ environmental ,ww4l*~~. solution, obviatethe need for’ the DETA due to aerobic oranaerobic UnionCarbide endDowsee.nnnertain proposedchemical fate testing of DEM. biode~adadonIs eossuary. With It. as to whether EPA Is proposingto Dow conrInd~i4hoerthese s~es that comments, Dow submitted a desaiption require qualitative orquantItative It is unlikely that an N-u.lfrosamine of an unpublished preliminary in vitro chemicalfsta.testlngof DETA ~ - derivative of DETA wiltform In aqu~s chemical study fu.~ by Dow det’i.ninb~gIf theN-rduosamlna solutions. or If It does form, It will aimed.at det~~rminlngif an N. derivativeo(DETA would be fcemad by decomposerapidly; Based on thebrief nitrosamine of DETA would form In mlaoorge”{~a.under aerobic and description of these studies which was aqueous solution when NO vapor was anaerobicconditions. Thefirma believe submitted to the Agency(no protocol for bubbled througha50 percent1 solution of that, with respect to aerobic condiioce, these studies or final study reports were DETAin D,O (used for-nuclearinagnetiC- the prutIimin~~yin vitro ~n1~aI ttlng submitted by the nwaufncturan), EPA - resonance purposes) under aerobic by Dow ~ur ~rAM believes that an N.rdfronins (- condli.ms. Uain~VItMYhOIM sail nuclear sufficientto stats that either.en 14- derivative aiDETA did, Infant fa~bu1

I Federal Register / Vol. 50, No. 100 I Thursday. May 23. 1985 / Rules and Regulations 21403

~1 decomposedwiderthe exper.mental developed for separating bound approximation for the production conditions employed. The fact that a N. diethanolamine (a close structural. of the N.nltrosamlne derivative of DETA nitrosanune derivative of DETA can, analogue of DETA) from N- fromDETA could be made by summing indeed, form In aqueous solutions Is nitroeodiethanolamnine (a close the exp.ected derivative production at all demonstrated by thestudies of Popp structural analogue of theN-rtltrosaxnine steps and assuming aerobic conditions (Ref. 14), In whichDETA was detected derivative of DE’I’A), and for detecting at all steps. For these reasons, the in waterby chemicaltraceformationof N-mtroeodlethanolamine in aqueous Agency will now require the chemical his substance to a N.mfrosamine solutions ate detectionlevel of I fate testing of DETA only under aerobic derivative, which was. subsequently nanogram/mI (1 ppb) should be conditions in the final Phase I test rule detectedby-polarography. The fact that. adaptable for use with DETA and its N- for this substance. theN-nitrosamine derivative of DETA nitrosamine derivative. These Thin.layer In contrast.tothe manufacturers of proved to be unstable in Dow’sin~Pitiv chromatographic methods have proven DETA. the Agency believes that the chemical systemdoes not Indicate that to be resistant to Interference by. - methodologies described In the studies it would necessarilybe unstable under contaminants present in environmental of Yordy and Alexander (Refs.? and 15) environmental conditions. Yordy and samplesused for chemicalfate studies. and of Pupp (ReL 14) and ths references Alexander (Refs. 7 and15) have shown of diethanolamine. In addition, the upon whichhis polarographie methodus that a substancevery similar In - Agencynotes that several investigators based (Refs. 18 and 19) do, In fact, chemical structure to theN-nlfrosammne (Refs. 20 and21) have published togetherwith other studies.(Refs. 18.20 derivative of DETA. N- comprehensive procedures for the and 21), constitute a sufficient model for nltrosodlethanolanulne, wasessentially detection and quantitation of a variety the chemicalfate studies proposed by stable to degradation In some ofN-nitrosamines in contaminant- EPA for DETA and are capable of environmentalwaters, especially during containing environmental samples, distinguishing between the chethical and the winter months, and was slowly which should easily be adaptable for biological production of an N- degraded Inothers, primarily dueto usewith respect to theN-nitrosarnlne nitrosamine derivative of DETA. The mia’obialmetabolism. In addition. Tate derivative of DETA. latterpoint is now, however, moot, since and Alexander (ReL 16) demonstrated With respect to thechemical fat. theAgency has clarified its position that - that three aliphutic N-nitrosamines (N- testing of DETA under anaerobic concern actually exists for thetotal -- nitro.odlmethylamine, N’ conditions, the Agency Is aware that production (both chemical and- - nitrosodiethylamlne. andN-nitr-osodi~n- denitrificationof thenitrites present in biological)of an N-nitrosanune - -. - propylamine). which may be viewed as the anaerobic,envlronment, thus derivative of DETArather thanjuetthi -. analogues of the N-nitrosamlne removing a necessary reactant for the biological portion of ~ derivative ofDETA, were resistant to formation of an N-nilrosamiae production. .. - - - degradation In soil, sewage, andlake derivative of DETA. may, indeed, occur. In summn~y,the Agency is cofitinuing - water. No degradation of these N- On the other hand, the presence of to require quantitative chemical fate. nitrosamines was observed in lake heavy metalsor certainpesticides in the testing ofDETA, using environmental - ( waterduring a 3.5-month period. Thus.’ anaerobic environment may inhibit the samples of lake water, sewage, and soil the results of Dow’sin vitro chemical denitrIficatlonprocess (Raft. 22 through under aerobic conditions. In the final study do not, In fact, obviate the need 24) so that a N-niirosamine derivative of PhaseI test rulefor this chemical for the chemical fat, testing which the DETA may still form. In addition. Bollag substance, and believes that adequate - Agency is requiring in the final Phase I et aL (Ref. 25) have shown that published methodologies are available test rule for DETA. unfavorable,environmental growth which, with minor modifications, will The Agency disagrees with the conditions, related to temperature. pH, permit the completion of this required manufacturers’ contention that no nitriteor nitrate concentrations, ledto testing In a timely fashion, without the methodology Is currently available for theaccumulation of nitrite, conditions expenditure of undue time andeffort. determining low concentrations of an N. conducive to the formation of theN. F. Necessity forMutogenicity Testiz~.of nitrosarnine derivative of DETA in nitrosaminederivative of DETA, under water, and refers these firms to the anaerobic conditions-in Isolated cultures DETA - polarographic methodof Popp (ReL 14), of soil bacteria. Thus, theN-nltrosamine Many additional comments were- - thedetails of whicham unpublished. but derivative of DETA might well be received on the proposedTSCA section which Is based (EeL ‘17) on the produced from DETA even under 4 test rulefor DETA in subject.areu published itudles of thepolarographic anaerobicconditions. However, the other than the five majorIssueswhich - - detection ofvarious N-nilresamlnes by Agency believes that the production of theAgency raised for public comment In Dahmeruatct (RILIO)and Omang and theN-nitrosamnine of DETA observed In that rule. Commentswere received from Hagtca~(Ref~.10), Although Popp -. chemical fate testing ofDETA under the major manufacturers of BETA In this (ReL 14) had demonstrated that an N’ / aerobic conditions would represent the country, Dow and Union Carbide, on the nitrosamine of DETA can, In-fact, be -~ upper bound for production of this gene mutation andcytogeneticstesting detected by polarographlctechniques, derivative under less favorable required in theproposed test rulefor - the Agency Is aware that contaminants anaerobicconditions.In addition, EPA DETA. These manufacturers believe that presentin the environmental samples to believes that most waste water there is convincing evidenceto be utilized In thechemical fate studies contpinIngDETA would be subjectedto demonstrate that DEFA doesnot have of DETA.may present difficultieswith sewage treatment processescontaining mutagenic potential in bacterial or - respect to the polarographic detectionof at least one aerobic step (more mammalian call systems, and, therefore, anN-nifrosamine derivative of DETAIn favorable to N-nifrosamine formation) they feel that the requirement for gene these samples.Should this prove to be before releaseInto waters whichmight mutation andcytogenetics testing the case, how.ver the Agencynotes be used for theproduction ofdrinking should be deleted in the final Phase I that the thin4ayerchromatographic water, and that for treatmentprocesses test rulefor DETA. . - detectionsystems which.Yordy and involving both. anaerobicand aerobic The evidence to which these Alexander (Ref~.7.and IS)have steps, or anaerobicsteps only, en upper- manufacturers are referringcoasletsof C

‘5 ~4I4~~ Federal Register I Vol. 50, No. 100 f Thursday, May 23. 1985 1 Rules and Regulations ~1 the results of mutarincity.testing. Ames SoimoneilalMicrosome Plate Test (strains TA-i535. TA-1537, TA-1538. sponsored e erby~Dow.orbyUnion was used withS typhthuzriwn stralni TA-ga and TA-100), as well as in the Carbide, which ~ .ia}v~ftt~dto the TA-1535. TA-1537.TA-i538, TA.-9& and yeast. Sacaharomyces carevi.siae (strain Agency by. these m,nufae±s~spriorto TA-100. BETA wasalso tested in this D4). under the conditions of the Ames in the pu’~safinn of ~ proposed tesl’ruie - system with the D4 strain of the yeast. vitro assay. On the other hand, the for BETA. and~which arw.-therefore. Saccho.romycescerevisioe. BETA was Agencymust point out that it does not discussed in theAgency’s tested overconcentrations ranging from - regard these negative findinge with “Thethylenairiamine Support Document” 0.01 to 10.0 ~slperplaje. both with and respect to BETA’s non.mutagenicityto a for that prupoud~sestrule. As described without the addition of a liver single bacterium and a single yeast as In the support document, thesubmitted microsomal enzymepreparation representativeof BETA’s complete studies consist oh (1) mntageaicily obtained fran. theliversof Aroclor- mutagemc potentiaL In orderto testing performed by Litton Bioneticsfor pretreated rats. The highest dose tested adequamiy assess the comp’iete Dow using theAmesSalmonella! - In this study, 10.0 gil per plate. mutagenic potential ofBElA. further Microsom. Plate Test and.5 strains of representing 9.5mg ofBETA. was gene mutation and cytogemcitytesting - Salmonella typà/auzthim, as well as the selectedbecause it producedtoxacity. will barequired. - D4strainofSaccb.aramyces carevisice, which was notclearly desudbed. in S Since the timewhen theme*agenicity with and without metabolic activation cerevisia.. Litton Bionetics (RaL 4) studies performedon DCEA for Union (ReL 4~,and (2) th. following tests reported that BETA was negative in all Carbide Corporation (Ref. 3) were - performedby BuaèyRun Research test systems, both with and without discussed in theTechnical Support -- Centerfor Union Carbide (Ref. 3) for metabolic activation,and concluded that Docnment for theproposed teatrule on each ofthreasamples of BETA (DETA- BETA was non-nuitagenic under the BETA. the Agency has audited, during highpurit3O BETA-commerciaL and conditions of the tests.- February 2810 March2.1982. the Busby DETA.hearts cut): (a)Anin vitro assay The essential agreement of the Run ResearchCenter in Export for specific Iocusmutation (at the negative results obtained by thetling Pennsylvama, where these studies were hypoxanthine-guanine . . of DETA in the Ames Saimonei]a/ performed. Therefore, the Agency’s pluosphoriboeyltransferase lecus)in Microsome Plate Test by theNational current conclusions with regurd to these Chinesehamster ovary(CHO) cells Toxicology Program (Ref. 26)and by studies were reachedfollowing an (with and without metabolic activation); Litton Bionetics (ReL 4) calls Into examination of the actual rawdata f~ (b) an in vitro assay for sister-chromatid question theearlier report by these studies at thetest facility.. — -- exchange (S~)in CHOcells (withand Hedenstedt (ReL 27) that BETA might scientific ~ between siwitlsts.. without metabolic activation); and (c) an have demonstrateda direct mutagenic who performed thetests at th. facility in vitro assay of the ability of the test effect in this test system. Hed.nstedt and EPA staff. as well asa’Arerlew -. substances to induceunscheduled (Ref. 27) testedDETA in the Ames and evaluation of the submitted thaly. deoxyribonucleic acid (DNA) synthesis SolmoneiIa/Microsome Plate Test using reports. -. ( (UDS)in primary cultures of retliver SoLo,oneio tj.phthizzrium strains TA.- Bushy Run Research Canterhas * cell.. EPAnotes that only twoof these 1535 andTA—100. both with and without conductedth vWo mutagmuimly studies test procedures, the Ames Saimonella/ theaddition of liver microsomalenzyme of the ability at three samples of BETA Microsome Plate Test and the specific preparations obtained from the liversof (BETA-high purity; DETA-commereiá locus mutation test in CHOcells, are rats pretreated with Clophen Mo, a and BETA-hearts cut)to induce spe~c tests for gene mutation perse. The tests polychiorinated biphenyl mixture locus matations in Chinesehamster for SCE in CHO cells and for UDS in similar to Aroclor. BETA demonstrated ovary (CR0) cells (Ref. 3). In thr primary cultures ofrat liver cells are a direct mutagenic effectin this test presence or absence of a niicrosomal useful asirulicators of geneticdamage, system, which was notaffectedby the liver enzyme preparation fromAroclor but they do notsubstitute for essays of presence or absence of the metabolic 1254-pretreatedmale rats (S-9 fraction). gene mutationper se and they do not activation system. However, the BETA the threesamples. ofBETA were tested addres. the ability of the test chemicals used for the testswas found tobe in this in vitro systemat concentrations to induce chromosomal aberrations contaminated with unidentified ranging from L25x10 ‘percent to (cytogeruc effects). No cytogenicity test impurities, which the author believed 40x 10-2 percent. The investigators dataare currently available for DETA. might be alkylating agents. The author concluded that none of the three BETA was recentlytestedin the concluded that BETA, or some samplesof BETA produced specific AmesSairnon.IZa/Mioro.omePlate Teat unidentified alkylating ImpuritiesIn the. locus mutations In CR0 cells under the by the Environmental Matagenasia Test BETA sample tested. may pose a - experimental conditions employed. wlti Development Program.-. part of the .nuutagenic and carcinogenichazard. or without metabolic activation by S-a NationalTo ‘olo-Program. awLthe / Based on evidence from thestudies faction. The Agency agrees with this- - investigators conclud.èthat BETA gave- conductedby the National Toxicology conclusion, and has, therefore, negative test results In Sal,noruneila Program,(Ref. 26) and by Litton determined that no further in vitro gene typhimrzrnzm strainsTA-ga TA-lao, Bionetics (Ref. 4), the Agencyconcludes mutation testing is requiredfor BETA. TA-1535,,and TA-lw. with or without that the positive results observed Itt the However, asdiscussed below, the metabolic activation by liver study by Hedeustedt (Ref. 27) were Agency I,requiring in vivagene microsorasi enzyme preparations probably due to the unidentified mutation testing ofBETA Inthis final obtained from Arocler-pretreated rats or Impurities detected in the sample of Phase I test rule. -. hamsters. BETA was tested ina series BETA usedfor testing. Bushy Run ResearchCenter has -of coacentantions r~~ngzngfrom33.ti gig Based on the data contained in the - conducted in vitro studies of the abilitl to 10,000 j4 perplate (Ref. 20). These Litton study (Ref. 4). u well as the data of three samples of DET&(DErA-hlgk results .ss~st~allyegreewiththoes - presented in the NTP study (Ref. 26), the purity OETA-cnmm~rciaLmd BETA- observed in a study performed~Litton Agency rnneludasthat DET&has been - heartLcut) to i~,rhie~ajster,.chrosialid exchanges (SCE) In.~in.se.ba~itir.- C Bionetica for ~ ~ _____ - . demonstrated to be nnrunut~gei~irIn the. Company ~RaL4). Zashi.ste4. the -.. bactarium~Salma la.4phimuthtm. - ovary(CHO) cells..bolh.Iu.tb.pr.s.nc

- -- - A

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and absence of a rnicrosomal liver With respect to the studies by than either DETA-hearts cut or BETA- enzyme preparation(5-0fraction) Slesinald at aL (Ref. 3) on the ability of high purity) failed to give astatistically— obtained from the livers ofAroclor 1254- three samples of BETA (DETA-high significant elevation ofliDS in this test treated ~ I~theabserice of S- - purlty DETA-commercial and BETA- systemmay be related to inhibitory 9 fraction, a statistically significant hearts cut) to induce unscheduled DNA effects of impurities present in this increase in SCE was observed far both synthesis (UDS) in primary cultures of sample of BETA. The conclusion that DETA’hAgh purity and BETA- - rat hepatocytes, the range of doses DETAelicits UDS in primary cultures of — commercial at thehighest dosage level tested was inappropriatelyselected. rat hepatocytes. togetherwith the tested (20.Ox10~percent). anda This dose range was selected on the conclusion that BETA induces SCE in simdarly statistically significant basis of toxic effects elicited by BETA CR0 cells cultured in viti’o, indicate that increase was observed for BETA-hearts in CR0 cells In the cours. of studies of BETA should be testedfor its ability to cut at the sarond highest dosage level the ability of BLTA to elicit specific induce in vivagene mutations in tested (1O,Oxlll”percent) under the locus mutations In these cells. Thedose. Drosophila melo.aogaster. asdiscussed range selected for LIDS testing should - same co~litions,An slavat.d thdd”~ below. - of SCE was observed at a DETA-heerts have beenselected on the basis of In summary, theAgency camsot cutconcentrationo(20X10”pemeat. DETA’s toxicity to the primarycultures adequately uses. themutagrmc but the increase was nct.st.tistically of rat hepatocytes used for ths LIDS potential of DETAwith the information sIgnificant. In the presence~ofS-S studies. not on the toxicity observed for currently available, BETA ha. been BETA in CR0 cells. In addition, there is fraction, a statistically signifiomit. shoasafReEs. 4 and 20) to be.- - - Increaseof SCE was en only for the. no indication in theresults presented for nonmetagenis to five sirains of the the LIDS studies that the highest dose of DETA.hearts cut sample eta dosage bacterium, Salmonella i4imer,um,- ‘ level 011.25 x10’peroenL DETA tested (1~0x1O”percent) was, in fact, toxic to th. primary cultures of rat and to the D4strain of theyeast. Dow and Union Carbide contend that hepatocytes used for this test, as is Socthammyes, carey/sloe, under the these results are not indicative of a recommended inboth the OTS test conditions of theAmes Salrr,oneila/ - - positive respoese because they occer at guideline (Ref. 28) and the Gene-Ton Microsom,PIateTest In addition. single dosage levels andno dose- Work Group report on this assay (Ref. ~tudie~exist (Ref. 3) which indicats-that response relationshipscan be 29). Nonetheless, statistically aigniflr~~nt DETA do,s net-Induce sped& toes.’ demonstrated. The Agency does not increases In UDS were observed both mutations in Chinesehamste’overy agreewith this contention. The OTS test with BETA-highpurity and BETA- (CR0) cells, but does induce~sister- - guideline for thein vitro SCE assay (Ref.. hearts cut at concentrations of chrorn.tid exchange (SCE) In’OfO’celIs. 28) states: ~‘Thereare several criteria for 0.01 x10’ percent and0.3X10’ percent enddoes induce nnscheduledDNA determining a positive result one of (based on DNA-bound radioacthnty3. synthesis (liDS) Inprimary cnlt~e,of which is a statistically significant dose- but neither sample of BETA elicited rat hepatocytes. On the other hand, the Agency notesthat no cytogenicity test relatedincrease in the number of SCE’s. statistically significantly elevated1 UDS Mother criterion may be based upon at a concentration of 0.1 x10” percent data are currently-available forBETA. detection ofa reproducible and DETA-high purity induced an elevated. The Agencybelieves that thepo5ttive statistically significant positive response UDS at o.ixio’ percent, but the responses which DETA has exhibited In foi at least one of the test substance increase was not statistically significant the SCE and liDS mutagenfcftyassays concentrations.”Since Slesinski ci a!. Nosignificant effectswere observed for indicate that this substancemay pose an ‘(Ref. 3) performedonly one experiment BETA-commercialIn this test system. unreasonabli risk of both gene for each of threeDE’I’A samples in the The Agency disagrees with Dow and mutations and cln’omosomal absence of the metabolic activation Union Carbide that. because the aberrations. notwithstzndIng~the . - system for SCE in CR0 cells, as well as statistically significant positive resposes negative results exhibited by this one experiment foreach sample in the In the LIDS assays of BETA couldnot substanceIn in vitro gene mutation presence of themetabolic activation dearly be shown to be dose-related. the assays in bacteria, yeast, and system, nothing can be said regarding responsesobserved werenot truly mammalian cell. in culture-, Therefore. the exact reproducibility of the four positive. The OTS test guideline for the the Agency is requiring both in viva statistically significant positive results in vitro liDS test (Ref. 28) containsa gene mutation andin vitro and in viva which occurred at single dosage levels. statement to the effectthat a cytogenicity testing of BETA hr thefinal However, sincestatistically s%nifi&~ant reproducible, statistically significant PhaseItest rulefor thIs substance. Increases of SCE were observed In respons. with respect to UDS for at The AgencyIs requiring that BETA be separate experimentswith thethree - least ons dosagelevel may be an - tested for in vivagene mutation effects. BETA samples Inthe absence ofS-~ Indication of a positive response In this utiH2ilig the-sex4lnked recessive lethal - fraction at concentrations ranging from’ test system. No attempt was made by assay in Drosophila meianogaster. Ifthe 10.0x10’~parceidto moxio’ percent. Slesinskl at a!. (Ref. 3) to verify the results are negative for BETA in thetest the Agencyconcludes that BETA reproducibility of the results of the UDS in Drosophila, no further in vivagene (constituting themajor fractionofall studies. However, because two of the mutation testing Is required. Ifthe three samples tested) hasexhibited a threeDETA samples~(DETA.highpurity results arepositive for BETA In the positive response In this in vitro system. andBETA-hearts cut)each gave Drosophila system, then theAgency is Therefore, no further in vitro testing of statistically significantpositive requiring that BETA be tested In. the DETA for SCE in mammalian calls is responseswith respect toUDS In mouse specific locus assay. Guidelines necessary, but, as dis~m~edbelow, the separate experimentsat concentrations for all of these test procedures have Agencyis requirlngin vivogaas-- of 0.01 xiO”percent andO.3xlr’ beenpublished by theOfficeof Toxic - mutation testing of DETA In Drosophila percent. theAgencybelieves—thatBETA Substances (Ref. 28). EPA is also mR!nni~gnIMi~.based. in.part, on BETAs must be regarded as an in vitro ~ requiringthat BETA be testedfor its- - positive response inthe ~. vitro test fo~ of UDS in rat hepatocytes. Thefact4hat - ability to induce both in vitro.aid in -. vivo r~Pnn1wJ~!tAJaberrations, usingthe ( - sCEIncRaceiis. - BETA-commercial(whichIi less pure - 21406 - Federal Register I Vol. 50. No. too / Thursday, May 23, 1985 / Rules and Regulations

test sequencesoutlined in Unit IV.B. of recessive lethal assay in Drosophila, mouse specific locus test and the this final Phase Itest ruM.~ . - -. andthis requiremenriscontained in the heritable translocation test), EPA has ( The generaisequencàof tiered tests final Phase I test rule for DETA. decided to utilize automatic triggers usually employed by EPAin assessing With respect to cytogenicitytesting, if between assays contained in Iower,tler the mutagenic(both. gene mutation and. a substance gives a negative response in tests, and a “presumptive automatic cytogenetic) potential of:ch~mfcal an in vitro cytogenetica assay, It is then thgger and opt-out” approach between substances, portions of whIch are tested in an in vivocytogenetics assay. lower-tier tests and end.point tests in required in this final Phase I test rule for If the substance exhibits a negative . this final PhaseI-test rule for DETA. DETA (see Unit IV.B.). have-been response in the latter assay system, then Under this approach, EPA Is previously described in proi5~isedtest no further cytoganicity testing is promulgating a tiered testing scheme for - - rules issued by the Agency for meóityl requiiet If a substance exhibits t mutagenicityfor DETA with automatic oxide (48 FR 30699) cresols (48 FR positive response in either the iii vitro or triggers to additional esutagenicity - - 21822), and ethyltoluenes, in v/va cytogenetics assay, then the testing (Inciliding thetwo end-point - - trimethylbenzenes. and Cm arom8tic substance is testedhr the do,nfrt~nt tests). Befor. testing is initIatedin one - hydrocarbon fraction(43 FR 23068), and lethal assay. A positive response In the or both of~the end.point mutageniclty are morecompletely described In the dominant lethaiassay indicatesthat the tests. EPA will hold a public-program final Phase Itest rulefor C, aromatic substance should be testedIn the ~ If theresults of theprevious tier hydrocarbon fraction (50FR 208621 May heritable traaslocation assay. A - tests are positive. Public-p.rtic2pation In. 17, 1985). Although these general test negative response inelther of the latter this pro~amreview will be’either In the sequences are usually employed, the two assay. indicates that no further form of written public comments ~a Agency ultimately specifies the required cytogenicity testing Is required forthe public meeting. Requestfor public mutagenicitytest for each specific substance. Since no cytogenicity test . comments or notification ofapublic che~calsubstance on a case-by-case data are available for DETA. the final meetingwill be published in theFedsial basis, With respect to gene mutation PhaseI test rule for this substance Register. If, after the review of public testing, if a substance tests negatively in requires that BETA be tested In - comments, no change In the test. the gene mutation assay in Salmonella. accordance with the tiered testing program Is deemednecessaryby ffl’A. It is then tested in the specific locus sequences for both in vitro and in viva testing will continue to th. next tsat~ mutation assay. In mammalian cells In cytogenetics. as described above. The without delay. EPA will provide- culture. If the substance tests negatively Agency’sresponses to commentsoe.the - notification to the test sponsor($that: In the. latterassay, then no furthergene tiered testing sequences for gene the next tier-test-should be conduct,d11.. mutation testing is required (in the mutation testing and for cytogenetics theAgency believes additional testIng Is absence of other positive mutagenicity testingmaybefoundrntheflnalPhasel nolongerwerrantedaseresultofthe test data). Ifthe substance tests test rule for the C, aromatic- - earlier test results, public comment, positively in either theSalmonella assay hydrocarbon fraction (50FR20682). scientificjud~nent,andother or thespecific locus mutation assay in Asdescribed in detail in the final appropriate factors, EPA will issue a mammalian cells in culture, then it is Phase I test rule for the C, aromatic proposed amendmentto “opt-out” by~~ tested in the sex-linked recessive lethal hydrocarbon fraction (50 FR20662). the repealing the existing requirement and. assay inDrosophila. If the substance Agency feels that there Is a consensus in after consideration of public comments yields positive results in the.Drosophia the scientificcommunity on both the on theproposedamendment. issue a assay, then it is testedin.the mouse need for, and themanner of, identifying final decision whether to rescindthe specific locus assay. Negative results In toamnialian mutagens, and that its - rule requirement. This approachoffers • either theDrosophila assay or the proposedscheme for Identifying these the advantage of allowing the mouse specific locus assay Indicateno agents is in keeping with’those incorporation of scientific judgment -. further requirements for gene- mutation recommendedby experts in the field of on theweight of the evidence testing. mammalian mntagenesis. Further,-while ~- after’ the initial testing tiers have been DETA tested negatively, both with It I. recognized that thereis, as yet no completed-andallowing change in lest and without metabolic activation, In generally accepted single methodology requirements to respond to specific Ames assays using both Salmonella for estimating human risk from- chemical issues, while i~otsignificantly typhimurium and Sacchoxomyces mutagenic agents, it is the Agency, - delaying-higher-tier testingwhen it is carevia/ce. and also tested negatively In view that appropriate methodologiesdo deemednecessary. a specific locusmutation assay In’ exist and are usable. Therefore. the- EPA’has decided not to use thepublic Chinese hamster ovary cilia In culture. Agency concludesthat it Is appropriate program review approach between the BETA would. therefore, normally notbe at this time to obtain mutagenicitydata . lower.tler mutagenicity tests for the . -. .ubject,Intbe.bsesceo!otherpositlve ~,nDETA with whichto perform BETA test rule. EPAbeltoves ~ use of mutagenicitytest data, to. requirement estimates of mutagenic risk for this automatic triggers between these tiers Is for furthergene mutation testing. substance for regulatory use, should suitable. It should be noted that this However, aspreviously discussed. the DETA prove to be a mammalian germ- does notexcludethe public froni’ Agency ha. concluded that DETA cell mutagen. - . requesting modifications-in the.test - - Induces sister-chromatidexchanges For reasons morefully described In program. Provisions are available wider (SCE) in Chinese hamsterovary-cells in the final Phase Itestruleforihe C. - section 21 ofTSCA for the public to culture and induces unscheduled DNA aromatic hydrocarbon fraction (50FR petition EPA at any time to amendanile synthesis (UDS)in primary cultures of 20682), EPA believes that theuse of undersection 4. EPA. Test Rule - rat hepatocytes. The positive results automatic triggers between the assays - Development and Exemption Procedure displayed by BETA In. these tatter two containedin themutagenicltytesting’ t~uIe.published In the FsdiraI Register of assays indicate to the’Agency that scheme for BETA-Is appropriate: October10. 1984-(49FR 39774). Includes- BETA should reenter the tiered test however, in en effort to Incorporate procedures for-Industry test s~Cnsor~to sequence for gene‘mutation testing at scientific judgement priOr to thi use of request icodificafions to‘testguidelfnes thesecond-tiarlevef,’the sex-linked the end-point mutagenicitytests (L.., the (but notto test requirements).” / Federl Register ,L Vol. 50, Ne. 100/ Thursday. May 23. -1~85/ RWer and Regulations 21407

Since the time at which the proposed comment arid to incorporate the. sp~ciiãc general population to a suspect test rule for BETA was published by the t... ~ iy,difled SY ~m-uiuiogefl.N-Nltru5udietttunolamine. a Agency (47FR183861 April29.1982). the a re~n1tof public comments) into the known animal carcinogen (Ref~8 EPA has adopted an approach of rmal Phase II test rulefor DETA as through 10). is formed under requiring tiered testing sequencesfor enforceable testing requirements. Any environmental conditions in waters - coatais~diedianolamine,a close both gene mutation and cytogeneticz- mod ficadna.oLthasa.raquAtemIUts testing which contain automatic triggers would require the Agency~sapproval structural analogue of DE’I’A (Ref. 7). for required chronic oncogenicitytestii~ and, when appropriate, publication of Chemical fate testing4 of DETA is when a chemical substance elicits the proposed modifications-forpublic required to det” ”lf~a.~. positive test results in certain of the comment. Comments from these firms nitr-osamine derivative of DETA can be mutagenicity assays. The mutagenicity were considered and addressed in the formedunder environn~s~talconditions. testing actually required for a given final ruleon Test Rule Development and The finding of potential-adverse health chemical substance. as well as the. Exemption Procedures. published in the effects as a resultof subd~ronicor selection of those mutagenicity assays Federal Registeron October10, 1984 (49- chronicexposure to DETA is based, in (if any) forwhich positive results will FR-39774). part. upon the stuRe,-of F8~ino(Ref. 2), triggeran automatic requirement for which indicate DETA-related adverse L Covfideótiollnforrnation Corned iii -effects on the liver, lung~and kidneys chroniconcogenicity testing. is - StudyProtocols - - - (and. possibly, the spleenand adrenals) determined on a case-by-case basis. Following carefulevaluation, the Comments from Allied on the ~ ~onica~y expo~dtoBETA. proposed rule for DETA were ~ This finding is also based. In part. upon Agency hasconcluded that such-triggers by the Agencywhich raised that firms the studies of Trubko aed T.~tyakova for chronic oncogenicity testing are - appropriate for DETA. Because the concern about the possible breach of the (Ref. 30). whiciidemo~e1ethat proposed test rule for DEI’A contained - confidentiality of proprietary exposure of rahintafor8 months to no requirement for onccgenicity testing. informatton which might occur in Phase DETA via the thinking watercan result either as an absolute requirement or as II of the test rule development process. ~ ~ ,~gn1ficamdecease in prothrombin in general, through the negotiation and activity and significantIncreases in the - a result-of positive test results in publication of detailed protacolalea - -- euivitisz .fs~n giutaru~ate-.oxalate specified required mutagenicity assays~ EPA is proposing elsewhere in this issue - requl dta.ta4oe BETA e,~orethe,~ nüramiiei~naglutami~Te-pyruvate substances which are thesubjects of - ~nsaminase The Agency ~ --- of the Federal Register under section - 4(a) of TSCA. that manufactweieend differentTSCA section 4 test stiles.. that there are insufficieht ~ - .,Allieds-corame~were-cvnsfdered~à’:~*~enee i!dch th~’effndi1mfth~ processors of DETA be reqtared to - - conduct chronic oncogenicity bioassays addressed in the final ruleon TesR~~ manufacture, distribution In~ie’ce. of this chemical substance,if positive Development and Exemption - - ~ use, or disposal of DVM er test results are obtained for D~TAin Procedures, published in theFederal any combination of such actlvities.on any of the following mutagenicityassays Register on October 10. 1984 (49 FR - hwnan health can reasonably be ,.~, - 39774). - ~‘ determined or predicted. and thattesting ( required for this chemical in the final ‘l Phase I test rulefor DETA (1)-The sex- IV. Final Test Rule for - - of DETA with respect to such effectsis linked recessive lethal gene mutation Diethylaeetriamine - necessary to develop such date~ - - assay in Drosophila melanogasLer. (2) B. Required Testing ,.“ the in vitro cytogenetics assay, or (3) the A. Findings in vivo cytogenetics assay. The EPA finds that the manufacture. The EPA is requimipg’~atBETA be processing. use, and disposal of DETk - tested for oral subefironic (90-day) - C. Role ofProcessors in the Testing-of may present an unreasonable risk of - - toxicity in aLleest one nmmmalian DETA injury to human helath due to potential- species. ~i accordance with theOTS He~ Effects Test Guidelines, Many comments were recei~&with - mutagenic. oncogen~.and.subchronic effectsof the substance for the reasøns - publishedby the National Technical respect to the responsibilities and Information Service (NTIS. P982— obligations of processors. both with presented in the proposed test rulefor regard to thespecific proposed test rule BETA (47FR 183861 April 29. 1982) and 232984), with respect to oralsubchronic more fully described in the support’ toxicity studies, testing of DETA in at for DETA and with regardto the TSCA least one mammalian species will be section 4 Test Rule and Exemption documentprepared for that pmpoaed’~ Procedures. In general. These comments rule. The finding of potential mutagenic considered sufficient,asopposed to the were considered and-addressed In the risk is based on thestudies of Slestuski requirement for testing in at least two et ci. (Ref. 3), which indicite that BEtA mammalian species, as presented in the’ final rule on Test Rule Development and proposed test rulefor BETA. Exemption Procedures. published In the,- induces sister-chromatid exchanges - (SCE) in CHO cells In cultureand - The Agency is requiring that a dermal Federal RegisteronOctober10, 1984 (49 absorption study be conducted with - FR.39774). - - - induces unscheduled DNA synthesis BETA in the same mammalian species H. RequirementofStudy Protocols (UDS) in primary cultures of rat hepatocytes. The finding of potential selected for oral subchronic (90-day) Rather Than General Study Plans oncogemcrisk is based on the testing. -‘ The-EPA is requiring that BETA be hypothesis that a N.mtrosamlne - Another issue relating to the proposed tested for mutageniceffects, both with test rule for DETA. which was derivative of DETA may be formed In environmentalwaters, soils, and gene mutation and cytogenetics testing. commented upon -by CMA andAllied and is requiring the following sequence - sewage. and may survive the treatment Corporation (Allied) is the Agencys for this testing. intentibn to publish proposed study of contaminated watereprior to their - plans submitted for DETA for public use for drinking water, thus exposing the IWNO ccci SMOISM ( /~* 21408 Federal Register / Vol. 50. No. 100 / Thursday. May 23. 1985 I Rules and Regulatloril

Testir~of ~TA for Ir~ucir~ In Vivo Gere ~tztatioRs Sex-linked rec~sive lethal. — ~~Negative — ~—~b further assay in - - - tastir~ Drosophila

-- - Positiv -

specific~ - 1OQJS assay -

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

In vitro - ‘ - ‘ l~further - - —~ —~ cyto- Negative In vivo cyto-- —~ Negative ~ testirç -~------genetics- - geretics - - - ‘. -- -... assay assay - - - :

~sit ive P:~sit ive

Datiinant lethal —-s--- :~tj~ - assay

------Positive

~rit~le translocat ion assay -

ccci

. (

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

The Office of Toxic Substances has Section 4(c) provides-that any person applications, and study plans, and to previously issued guidelines for all of - required to test may apply to EPA for an conduct testing. unless manufacturers the test methods mentioned above (Ref. exemption from that requirement. EPA’s failto sponsor the requiredtests. The basis for this decision Is that - -23). - — ------final regulations for the issuance of The Agency Is also requiringchemical exemptions from testing requirements manufacturers areexpected to Indirectly fate testing ofBETA In environmental are in40 CFR Part 790. In accordance pass thecosts of testing on to processors samples of soil, lake water;and sewage. with these regulations~any through any price increase of BETA. under aerobic conditions, following the manufacturer or processor subject to a F. Reportfr~gRequirements general methodology utilized by Yordy Phase! test rulemay submit an and Alexander(Refs. 7 and 15)and Tate application to EPA for an exempfloe.. - EPAisrequirlngthatalldata - and Alexander(ReL 18). The final - - from submitting itudy plansand from developed under this rulebe reported in requirement-foechemical fate testing of conductingany orill ofth. test accordanc, with theTSCA Good- DETA has changed from that appearing requiredunder such aavis. If Laboratory Practice (GLP)-standards In the proposed--test rule-foe BETA in manufacturers perform all the required - which were published In 40CFR.Part that no anaerobicchemical fate testing- testing. processors will be granted 792. These final- GIP standards apply to Is now required. - — • : - exemptions automaticallywithout- : this rule. - - - having to file applications. - - EPA Is required by TSCA section CTessSabstance ““ - Manu1aa~-~andprocessors whoare - 4(b)(1)(C) to specify the timeperiod EPA is requiring that arelatively pure subject to th, testing requfrements of - • during whichpersons subject to a test grad.ofBETA be used., the test this rulemust comply with thetest rule rulemust submit test data. These substance. A purity of atleast 99 development andexemption procedures deadlineswill be established In the percent is specified In this rule. BETA of in4OCFRPart7gO. - secondphase of this rulemaking In this purity (DETA.High Purity) is EPA Is not requiring the submission of which study plans areapproved. The commercially available. equivalence data as a condition for procedures for the secondphase - - exemption from therequired testing. As - rulemakingare described in 40 (~RPart D. Persons Required to Test noted above. EPA is Interested in Section 4(b)(3)(B) specifies that the 790. evaluating theeffects atutbutable to ‘~SCAsection 12(b) requireithat activities for whichtheAtlminiatrator - - DETA itself, and has specified a persons who exportor Inten~b~ert makes section 4(a)findings relativelypuregrade substance for - to aforeign country achemicgl (manufacture, processing, distribution. testing. use and/ordisposal) determinewho substanc,or mixture for whith~ bearsthe responsibility for testing. £ Test Rule Development - submission of data is requiredunder. Manufacturers are required to testif the Development of this test rule-for - section 4. such as BETA,notlfy EPA of findings are based on manufacturing. DETA will be a two-phase process, In such exportation or intent to export. distribution, use, or disposaL Phase I. this test rule is being While the results of required testing may “Manufacture” is definedIn section 3(7) promulgated for BETA specifying not be available for some time, a notice of TSCA to Include “import.” Processors- certan health effectsand environmental to the foreign government *bout the are required to test if the findings are fate characteristics for whichtest data export of BETA serves to alert them to based on processing, distribution, use, or are to be developed. In Phase II, the Agency’s concern about the substance. It gives these governments disposal. - following promulgation of the Phase! Because industrIal workers. test rule, those persons subject to the the opportunity to request such dat&that consumers, and thegeneral population rulewill be required to develop study the Agency may currently possess on may be exposed to DETAduring plans for thedevelopment of data thesubstance, plus whatever data may manufacture, processing, use and pertaining to the effects and becomeavailable as a result of testi~ disposal, EPA Is requiring that persons characteristics specified In thePhase I activities. Thus, upon the effectivedate who manufacture or process or who rule. Within 30 days from the effective of this rule, persons who export or intend to manufactureor process this date of the final Phase I test rule, intend to export BETA must subnd~- chemical from the effective date-ofthis manufacturers mustsubmit toEPA a notices to theAgency pursuantto TSCA test ruleto the end of the reimbursement letter stating their intention to sponsor section 12(b)(1). Foradditional - period be subject to therule. Theend of testing or an application for exemption. Information, see49 FR 45581 (November thereimbursementperIod.willbe 5 Test sponsors mustsubmit their study 19, 1984). yearsaftarths d.a~1nefor submitting - plans to EPA within 90 days from the TSCA sectIon 14(b) governs Agency the last final report under thePhase II effective date of the Phase I test rule. disclosure of all test data submitted test rule. Asdiscussed In theAgency’s -- Afteran opportunity for public pursuantto section 4 of TSCA. Upon feetrule developme~-aadexemption / comment EPA will promulgate arule receipt of data required by this rule. the procedures (40 CFRPart 790), EPA adopting the study plans, u-proposed or Agency will publish a noticeofreceipt expects that manufacturers will conduct modified, as the test standards and in the FederalRegister as requiredby testing and that processors will schedulesfor BETA for thetests section 4(d). Test data received pursuant ordinarily be exemptedfrom testing. requiredby the Phase I rule, Testing will to this rulewill be made available for BecauseTSCA contains provisions to also be subject to EPA’s generic TSCA public Inspection by anyperson. except avoid duplicative testing, not every GLP standards. Persons who submit the In those cases where the Agency personsubject to this rulemust study planswill be obligated to perform- determines that èonfldential treatment - individually conduct testing. Section the tests In accordance with thetest must be accorded pursuant to section 4(b)(3)(A)of TSCA provides that EPA standards and schedules developed. 14(b) of TSCA. - - - - may permit two or moremanufacturers Modifications to the adopted study C. fnfonement Provisions - - or processorswho aresubject to the rule pla~scan be madeonly with EPA to designate one such person-or a approval. - - The Agency considers-failure to qualifiedthird person to conduct the Processors will not be required to complywith-any aspect- of a section 4 - - tests and submit data on their behalL submit letters of intent exemption rule to be a-violation of section 15 of ~ - 214i.~ Federal Register I VoL 50, No. ~o0 / -Thureday. May 23. 1985 /— Rules und &e~ilañ~

, TSCA. Sectioa 13(1) of TSCA makes it penalties of up to 125LQ00 for sach. day of On the basis of this study, theA~ncy - - - unlawful for any personto failor refuse - violation and imprisonment for up to - - believes that therewill be available test - - tocywithany~eorc~iuued one year. Other remedies areavailable facilities, and persormelto pezfor~the - - ‘uñdcl~ection4.Sa~fon15t3la1TSCA - to EPA under sections 7 end 17 of TSCA. testing required in this test rule. makes it unlawfulfor anypersen to Sail such asseeking 1n injunction to restrain -- ~.r refuse to: (1 ablle~ormaintain violationa-oITSCAs.ction4 - em - - - te~o~~) submit repo~.a&fces, or Individuals as welt as corporations EPA has established a puk~Acrecord - - - other information, or (3) permit access tQ could be subject to ~ ~ for tins rulamakEig (dochatnumber - - or copying of records rrqui~edby the Sections 15 sadi&o1TSCAappl~te OFrS-42012B)u~hichisavad~ Ice Actor say regulation issued under any person who vio&~tesvarrouc uaspectmo. is. ikeOP7S liesdtng Room. TWA provisions of TSCA~.EP~~y, ~ Jtm E~-1W.4t~1.M Strest SW.. - - Additionally. TSCA sectico 15(4t discretion,pcocwi ~ ~ Washiogtoti. D.C..from & a.m. to4 pm.. ------makes it uilawful for any person. to fail as well as comp*o~es.themseises. In ‘-Monday through Friday. .ureØ legal -- ~-orrefuse to per~t4~ entry or inspecti~’oas particular, this ixhules inàvidasls wbá -thbliayi.Thnr~sd Inchei besóc - ~requiredbyse~~ ”ii.Section11 - report falseinformation or ~ ~ mtutma-bco theAgeuq ~ us applie, to any ‘establishment,facility. to be reported. heSiMS~’Is.the - -.: developing thi, proposal, and - or other premisesin whichchemical ,ubnusmonof f~Jss.~ or a~propnateFndersl Register notices. substances or mixtures are frau~,otstatements u~icn_co l’he A3ency will supplement therecord - ; manufactured. processed.stored. or held under isU.S.C..1001. - - - with~ddiba~alEforusetion—~Is ‘~beforeor after their distribution in , ~ receiveLThs-g~dsow the - commerce., . ,“Tbe Agencycona~en V. ~ Analymaolivia foilowi~~g - a testing facility to be a place where the To assess thepotential ec~- ~ ~ r~..~ chemical is held or stored and. of tbm fmat Phasu Itest .~ . -- ______therefore. subject to inspection. EPAhas prepared an e~oeunc - (1) Federal Registeraofl~pwiaiute, ------Laborctory audftslinspecdons will be evalso~that ~ ~ of -~tothis rule. consisting oft conductedperiodically in accordance the required testing cod coutyzes-lcor - - :(~)Notice of final4 rule on BETA. with the ~rocedvres outlined in TSCA. iuarke*~ t~~ jb) ~‘~~v~”p” rule on E~TA section 11 by da~natedrepresentatives (1) Demand-s j2)cost ~ (4~’FR 1~00J. - - of the EPA for the purposeof charactenatico. (3)~mi~ysti~ture. - -~ - (u)Nolice containing thefTC - determining compliancewith thefinal and(4) market expectations -~ desipietion ofBETA to thePriority List - rulefor BETA. These inspections may Based on a total ~ cost of - (48 Ffl 2J138~. -- be con~Zedfor purposes which $220,400 to$487,200 and an annualized ‘- - (dtNotice of !nal rule or EPASTSCA Include verification that testing has cost of $57i15 to $t~,ZS3.the~umic Good LaboratoryThnctice Standards (41 begun, that schedules are beingmet.- - evaluationofDETA indicates that the FR 539~). - that rapbrts accurately reflectthe potential for ~ economic effects (ci Notice ofSnal ruleon test rule - - under~~raw data and interpretations due to the~etheatedtesting costs.is to~~ devnlopnieiit and .iou procedures and evaluations thereof and that the This conclasion Is basedon the --~-- (49FR 3~774i. - ~ studies are be~conducted according following obserratlon~til The demand (fl NoticeoflanE rule con~1~nfngdata - -- to the TSCA GLP standards and the test for BETA is relativelyinelastic due to -- reimbursement(48 FR 317551. ------standards established in the second limited potential forsubstitution in and (2) Support documents, consisting o~ - - phase of this rulemaking. — uses. (2) themarket expectations fc~ - (a) Diethylenatriamionsupport -- - - EPA’s authority to inspect a testing BETA aregenerally favorvbin~and (37 document. facility also derives- from section 4(b)(1) the relative magnitude ofthetest co.Us (b) Ecovo~napactana3ysiaoLlisoi - ofTSCA.wbichdlr,cts EPA to minor, I.e., an esthuatad 0.49 cnt per test rule for BETA. - promulgate standards fur the - pound to the upperhumid. This (3) Commusi~u~conmati~ot - -- developmentoftest data. These represents 0.31 percent ofthesafesprice (a) Written pebhe ~ standárdaansdefledins.nd~3~12J(B~l of BETA. The economic analysis - - - (l~3SemaarIs~of telephone - of TSCA to Include thoserequirement. presenting these conclns’n~~is inthded~--coi.~svstioei, necessaryto sui~ethat data developed- In the pubtic record for this rulemaking. ~ M~s~gs~*sn under testing rules arereliabl, and VI- ~ of~ ~ (d) Repode—çabI*sbed arid - adequate. and such otherrequfremsuis as are aeceumy topioetdsmeh Pursc& - unpualmbad lactoni materials, ki~~ — assurance-The A~cymafatafes that on )~1)a requires EPA (4) Test protocalfin-s .4——I laboratory inspections arenecoemry to to consider ~be a ions ~ absor’~.taèv of provide this usursncs. availability oldie facilitiesand ---- - Violators of TSCP~are subject to personnelneeded to perform the testIng ~8•References criminal and civil liability. Persona who requiredunder thenil..” ~efore. EPA- :~1i);N~H,Natisndie~, for submit materfaflymisleadingor false -- conductedastudy to assessthe ~ ~ sassy~g _____ - Information In connection with be availability of test facillliss and -‘~-- -- - Occupational--HasardSarsay DataBus. requfre~entof any provision of this rule personnel to harxfle the ~d.4DkmiI Washington. D.C. U.S. Depsr~ititH..hk may be subject topenalties calculated demand for tasting servicescreated by Education aniWelinw. ~.. - as if they had neversubmitted their section 4 testrules and tint programa (2) Fuilno. M. “~icperhiieiitaIStiuti.s on th — - dath. Under thepenalty pr~ioa of of __ __ section is ofTSCA. any person who - violatessection iScorid-be subject to a emi , - esting ustry Prolile of Itos.fI’ransmint by civil penalty of up to $25.DO0 perda~for ToxicologicalTesting, October. 1981,. - Service for D~C each violatIon. r~tanffo~ofviolations can be obtainedthrough, theN~Sunder - -- ~j sti~ai.p.s,-G~&.~sw..C~ - couldlead to the impositløn atorfminaI publication nuinber PB $2.-44lll73.~ - P.J.. anéHm W.C. “Uis*hy~mme Federal Register / Vol. 50, No. 100 / Thursday,. May 23. 1985 I Rules and Regulations 21411-

witjo Mutagenesis Studies: 3-TestBattery.” Determination of SecondaryAmines.” Z. prescribed for DETA is lees than Project reports 43-90.43-na. and 43-120. Anal. Chen. 185.161-174. 1962. (Translated 8128.243 overthe testing and - - Bushy Run ResearchCanter.19t0. (Subieltted lot EPA by Scitran). reimbursement period. Second, because to Union Carbide Corporation). - (19)Clang, S.V.. andHarrington. G.W. the coat of the required testing will be (4) LItton Blonetics,-Inc. “Mutagenicity “Determinationof Dimethylnifrosamuw and distributed overelarge production Evaluation of8—314 (DETA) In the Ames Nitrosoproline by Differential Pulse volume, the rule will have only very Soimonello/Mici’o.oare-Plate Test. Final Polarography.” Anal. Chem. 47(11): 1857- minor-effects(less than 0.31 percent a Report.” 1971 -(Submitted to Dow Chemical 1060. 1975. - Coinpeny). -- - - (20) Fan T.Y,. )CrulL I.S.. Rosa. R.D.. W0IL year) on producers’ costs or users’ prices (5) Ayanaba. A,. andAlexander. M. M.}L andFine. D.H. “Comprehensive for this chemicaL Finally, taking into ‘l’ranaforinetions Of Methylsniines end - Analytical Procedures lot the Determination account the natureof the marketfor this - Formation ofa HaxardousProduct. - of Volatile andNon-volatile, PolarandNon- substance, the level-of costs involved, Dtinethylnitroeemine. a Carcinogen and polar N-Nitroeo Compounds.” In and the expected nature of the - Mulagen. in Samples of-Treated Sewage and -- -“EnvIronmentalAspects ofN-Nitroso mechanisms for sharing thecosts of the Lake Wrter.~J. Environ. Quit 385-I9. 1974. - -‘. Compounds.” Walker. L.A., Grictiata, L, required testing. EPA concludes that (6) Aynniibs. A,,Veretrasta, W., and - Caste~nird.M., and Lyle. RI.. eds. IARC - Alexandcr, H, “Pormatiàn of , SCientific Publldatlons No.19. Lyon,Francs there will be no significant adverse Dtm.tky1nlb’oumi~ie.a Carcinogenand ~.RC 1971 economiceffects of any type u a.result Mutageu~isSoils Treated wIth Nitrogen - - (21) luenbarg,P. “AnalyticalMethods for of this rule. ‘ - - Compounds.” Soil Scl~’Soc,Amer Plan. O~S1flh1isL ~ “N-NZttVSO Compounth.’ Thiaregulation was submitted to the ~ iqpu, Scanliri. LA., *n(Taiu enbeam. S.R, ida. OfficeofManagement and Budget (7) Yordy) and Alexander lit “Formation ACS Symposium Series 174. Washington. (0MB) for’review as required by - - äfN-Nifrdtnditthanolamln.from - D~C.AmirecanChemical Society, 1981. Executive Order12291. Any comments Diethanoinnilne-In Lake Water .-‘4 ~i,yage,~’ (22) Bollag, ).-M.. and Bsrsbesz, W. “Effect ‘J, Ent,ftctm, QusL10.26e-271~ - ofHeavy Metalson the Denftrificatlon from 0MB to EPA, and any EPA - - (8) Drutkre~?,H, Preussinano. 3.. - Process in SoiL” J. Environ. Quit atlas-mi. response to th~necrunments. are - - Ivankovic.S., Schmahl D.. Aficham. J.. Slum. ~ -- - included in the public recordfor this G.. MenneLH,D., Muller. lit, Propoulos. p,, (21) Boilag. ).-M..and Henninger. NP.t nil.. - - - and Schneider. H. “Orgsnotrop~carcinogen. Influence ofPesticides on Denftrlflc,?don in B. Re3uIotory FlexibihtyAct Wirktingen hem 65 vemuchiedenen N. Soil and with an Isolated Bacterium. j. Ntti’oeoverbtndungen andBD.Ratten? (In Environ. Quit MS-lI 1975. - Under theRegulatory Flexibility Act Gernianl Z~Kribefóracb.09’.i03—201. ltd?. (~)~ S.. Watanabe, L. Honma, M., (15IJ.S.C, 601. -Pub. L. 96-354, September - (9) Hunch, j., Schmaltz. L, andHoffman 1). ~ Honda. S “TheEffect ofPesticideson “Effects of N NitrosothethanolammaandLi ‘theDenitriftcatlon In Paddy SOIL”Soil Sci. 19.1980), EPA is certifyingthat this teat - rule, if promulgated, will not have a- - Diethanolhydrazume Id Syrian Golden - 4lItt Nutrition 1~15-23.1954. Hamsters.’ Cancer Letters4.55-80.1975. - Bollag, 1.-lit. Orcutt, M.L and Bollag. - signtflcant impact aria substantial (10) Pretiumana, L Nab.. M.. liabs, H.. - ~. Denitriflcatlon by Isolated Soil Bacteria nujnberof small businessesfor the d Schin h1 ~ “C~In - - - f- ~ - - under Venous Environmental Conditions. following reasons Nitroaodiethanohamine in Rats at Five - - -‘ -- - - 1.-Based on the’Economic Impact - DifferentDose Levels.” ~ ne.. 4na107- (25) NTP. National Tozmilo~rPro~ani. Analysis preparedforthis rule, there Is ~ 1962. - - - - “Sslmonella-T.stthg Results on - only. one small- manufacturerof DETA - - 11 Boil - - and-ti S. “N’ Dlethyleneu’iamine. 1964. - - that-~mm~iufacturuyless than 00~ and Nitrous.Oxide A~cumuld” - an re5ca -- percent of the-estimated annual Denitriflcation in the Presence ofPeaticadi - “Mutagenicity Screening ofIndustrial £ ~ - i.~ ~ - Clmemicala Seven Aliphatic Amines and One domesticproduction of BETA. Although llVa i~5. ..y~ Vi~Ofl. tO - £ d 1’ eA - 1. 0 1 iI 1 39-845-849 1980. - i a as in a one a, crosom no figur,s are available to indicate - - . ,. Assay.” Mutation Re.. 53.198-196.1975. ~12) oeuacn, i~i.n.,aria imeuje. J.M. ,..m,tlc .. - - whether or not thereare small ~ i .~ ~ - - ~. -- j.~jmCeOitoxic nuustancas. businesseswhich import DETA. the - pane on or ono i - Nitric Oxide.and Nitrous Oxide durins - - total amount ofBETA imported Is V ~ITt - A - - 8 01. ArlIngton. VA. NatlcnalTechmcsl - - - -- C en ni ca on. .-.~, viron, - ~ f - - 0.4 ~. ~ estimated to represent less than I - MicrobioL 42:1074—1064 1961 - 1982ci -- o. percent of theestimateddomestic - - - - -(13) Stanier. R.Y., Melberg, L.A., and - - - 2129 ~u$uaL ’Ca~ - production of BETA. Thus, the ‘ I ‘ M.L.. el-al. ‘~Unzcbsdu1edDNA Synthesis estimated number of small - - Engiewood Cliffs. New Jersey: Prentice-Hall. Tests: a R~poflof the U.S. Environmental manufacturers (Including importers) - Inc., PP~~ .. Protection Agency Gene-Tax Program.” ~ ~ - - °n - - Mutation Res.1~383-410.1963. affected by thi. rule will be quite small. BlodegridabditycfPidyammass.” “~ (30) Trubko~E.I.~and Teplyakova, L.V “A 2. Small manufacturer, andsmall Del. 1t310-311. lV7a (Trsñaiatedfoe EPA by Study of Di.thylanetriamlns for Purposes of processors are not expected to perform UteratureResearch (pony). - Hygienic Standardization in Bodies of -- testing themselves, or to participatein (15) ‘t’?~tIY,J~R~ ~ ~ - - Water.” Gig, Smut. 37:103-104.1172.. theorganizationof the testing effort. “Microbial Metabolismof N- - rr~ I ~ hv lãM.a~stuyuResearch 3. Small manufacturers and small Nltrosodlethanolamltls in LakeWater and’ for EPA). - processorswill-experience only minor Sewage.” AppL Environ.MicrobiaL 39:559— ------costs, Ifany, In securing exemption from 563. 19~ - - - -.. - IlL Other Regulatory Raquk-~~~’-ET1 w361 235 m411 235 lSBT - -- (16)-Tate.RL,-IIL.andAlexand.r.M. - . - testing requirement.. - “Stability ofNltroeemmea In Samples ofLake ‘~‘ ~ “er - 4 Small manufacturers and small - - — Water Soil, andSewage.” J Nat. Cancer Inst. Under Executive Orderi~, ~A processor,are unlikelytobe affected by 54.327-330. 1975. must judge whetheraregulation Is - -- reimbursementrequirements. - - (1.7) Popp. Kit BASF. AG. Ludwiphafemm. “major” andtherefore subject-to the - - - .-- Germany. lattertoLL laCks. ~ re~uirementof a to In ct - - C Paperwork ROduction Act -~ :- - -- ChemicaisAssesamentDivisi~Office of - . -The Informationcollection Pesticides andToxicSubstances. U.S. s,ua.ysls. ~ ~u~u0U tor Environmental Protection1~gency chemlcal4ubatance Is not maor because requirements containedin this rule have Washington, DC W480. 1984 (March 13) It does not meet any of theaRena set been approved by the Office of (18) Dahmen. E.AJ.LP. Vader D.. and Van forth in marion1(b) of theOrder First Maimagemint andBudget (CluE) under - ( dirLaarss. j.D lime Polarographic theactual annualcost of thetesting - - - the prOvisions ofthe Papa~rk - - - Fsdor.~R~atss / Vol. 50. I’~.100 j Tbm’edsy. May 23, I8~5f Ru)es and ge~u1etfnrm,-

ReductionMt cL18~.4* U.S.C. 35~1.s Test Gsid.’~in.~far ~ isa~lcgy. the OECDCouncil on May12. 1187, med / seq. sad have hens a ead as ad~tedby theOECD Ceencd-cn the Pesticide Assessment Guidelines. mrmbe 2~-~-5~ May 12. 1981. and thePesticide published by NTIS (PBa—is~sto). Asee~montGuidelines.publishedby Additionally, thefullowing rei-hre~ List dReb~ecteIn 48 CP~Pt~II NTIS (PB 8~-1~. should be ccnsulte* Testing.Envir~antsLpsof~~n. (2) Mutaganiceffes—romc!~’?iJii (A) Feithuem R3, and MeThadi. HI Hazardous materiaL~en~- aberrations.—(i) Required testing. (A) ‘Ab.w~itIuiaofSomeOrganic Compounds An so vitro qtogonetkis lestMzsli be throughtheSkin in Man.” j. Invest. DerrostuL J.A. M~. conductedwithDW. 54:399-405.1970. - (B) An in v vcc geneticslest sheS (B) Feldman. 0.fthaó.P1L ~ ~.d be cenductedwfthD~TA.ftheievr~v “Percutaneous Penetration a(Steroidsin. Tozic54~—- - - cytogenetics test w~..j.~tedpwwuautto Man.” J. InvasS. DermsSaL52~4t.1169.. PW 78*4MRN~IDI paragraph tcX27~1!AJofthis ~sctLon’ produces. a negative resuli. (d~Chlfar,trsring—(~) Thesafbrs 4~~RPart7~is~“~‘ed (C) A d minararethal assay shaU be RequiredtsstYng. Testing to assess-N- as follower conducted with DETA, if either these nitrosamine formation. resultingfrom 1. Theauthuilty cft’ttlan farP1st 748 vib~cytogenedosteatecndce~M aerobic bLoIo~ndanid(orchemical continues toread as follows: purm~tto pez~sph(eX2~iXA)of thte transformeloc. aha~be camduciedwith DETA using environmental of AutSorlty 15 U.S.C.~O3.2811.~2L sectice produce. • positive result - (D) A heritable franilocation asmy leira ws2e~sewa~end so~ 2. By adding I 7~35~5~ send en shall be conducted with DETA. ø’tbe (Ii) Slec~ypz5mw.Forguideswe In follosen dominant lethal easeyconducted preparing study plane.thefollowing 5788.1879 pursuant to paragraph (cJZfi~C~of this. references should be conaultadi section or the in vfvo cytogenetics test (A~Yerd~r.J..~Alesaa~r,St Dlsthyls.strtem8is ~ETA) conducted psn~t to peregraph “Forv~~d~.56uesed6~aaoiamtee’from (a) Identification ofchemical test (c)(2U1)(B) of ~s ,~lsmprc~ Di~m~~-~Le~’Wale,end Sewage’ substance. (1) DIs*yIene~tamien(CM positive rendi. J.Environ.Qeat1~sb.~ 39ss. No. 1Th-40-O. alec known as DETA) (ii) Studyplans. For guidancefat (B) Yerd~J.. ~AI~mdi~14. ~eh6of shalibe kstsd~accordance wit,.this preparing study plans. theOTSMesith Metabo~acfN-Nftroeodlethenofo~ele Effects Test Cdde~nesfor thso~.omal Labs W~av4 Sewege’AppI. ~ (27~tslemfnr efat least effects publishedby NT~(PB ~- 232$84),ahonldbeconeultnd.Addaismal (C~Tate.LL.98. endAI~de~ar.- patent parityshs~be used as the leaf “Stability alt~ommine,In ~es ofLobe substances In aS tests. guidance may be obtained from ~ Water. 504. endSew~.”7,. Nat.Caesar ~ OECD Healthabets Test Gd~1iaII-P / . (b) Parsons ~ 54327-339: W~ - for genetintsmicd~.en adopted bythe (D)Poppfit”Sedfe,eethw plans.All pers~conductwliotestsea...Jnctt~r~andsubmitd~or prec~- OECD C~dJen htoy IZ 1803. end the 3Iod~m~tye4Fo*yaiom.7itn~ dletkylenetnieminefromJuly 9.1989. to Pesticide Asse,,nent Galdelines. Dot. 14310-311. 1977. (Translated for EPAby the end oftherebebiesoment peafod published by NTIS (PB83-153911). Llteratue ResomthCompeny). shall submit letters offz~essttc tent. - (3) Subah,vnfc effecLs—(fJ Reqeked (E) Dabeen. !.A.M.P~Va~D.. sad Yen exemption appl~atiuus.end study plans testing. A ninety.day oral subch,onin der Lames, l.D~“‘The Pelmugraphe and shall conduct tests and submit det~ toxicity teatshall be condintortwith Detenninarien of5ecmulsry~A~a~~ DETA in atleent one aaen~elien Anal. Chem. 186~181-174.1982. Cflamheed as speciSed In this aiitTui. Subpart A of for EPA by Sctfran)~ this part and Part 780 of ubapter s—es. ___ (li)Sterlyp.isns. For guidance In (F) Chaa~.S.V.. and ffarr~on.G.W. - (Test Rule DrseIIl~WI.IIZ and !xemptiOu “Detenmnientfce of VtmethyInutr~inesad Procedures). preparing s~fypisea the073 Health Effects Test Gddeheeefor ore? Nitrosoproilne’ by OlifereotfalPathe (Approved by theO~sof~ sad Polamgrvpby.” Anal. Coon. ~~—ra~ subchronic testing, published by W1’IS 1975. — underomtrof ammibea ~o033) (PB ~—232984J.sbsmM be cunsullet (G) Fan. T.Y. ICruli. LS..Rose. LlL.Waff~ (c) Heaith-effe~testing—ti) - Mdltional fi4aim,se mey be obtaiced Mit. andPine. D’.H Comprshanste. Mutager~ic .ffáte—~nesae~--~)- from the 0~ Health filecteTt Analytical Proceduresfor theDetesafentlen Required /~~ii.g (A)A een4~nd Guidelines fororal s.bchn~etitag~ of VOlatile analNon-volatile.. Polar eset Noo— recessivelethalteethe~’~ as a4.,yled by the C~ r~—~en polarN-NIatvan Compe~e.’be melanogenter ehaR besen~InL~with May12. 1~.and thePesticide - “Ea*oaniectal Aspects af~Nftroeo - DETA. - Assessm~Geddelteea psblkhed by Compounds.” WaScer. LA.. Gd~le.L.. (B) A apc~a ep~yehe~ Casteguaro. M.. andLyls. RL.ole. [ARC 5denWitPubVdsflxsNo.10.Lpen.Prancm be cond~a4with DErA. ~thS sea- - linked r~cuWmlethallest in testing. A darmalabsorption testshall (H) Issenheag. P. ~‘AnaIytfcaiMetbed, ten Drw$ilamalaaogasterconducted be condect.ôwithDETAIn theu Nltnvsamines.” in N.NItxoee Cnpaeadi puasnantto paragraph Lc~*I~A)of mammalian speciesused for the oral Sanian..R.A.. andTanneubaun.SR. .di. a.ctloo ~ntl”oms penitive penalt. subchronic(aO.day~testconducted ACSSymposhen SerIes174. Weahisgtes. (U)Stafyplazs. Forguldancele pursuast to p..,.,...,.h (cPi~b)of D.C- American Comical Society.11g. - preparing study pleas. the 01’SHealth section. Effects-TestGuidelines for gene (ii) Stsdyp?ess.Per guId~ (e) Avoilability~‘testgu~*’iñwe~7h. mutation assayspublished by the preparinigatady . theOlS Hndth 01’S HealthEffects Test Guidelines National Tethni~~aII~ SaevIce Effects Test ~ fan ~ cited in this final rule axe available &~ (NTIS) (PB 82-EZ~M1.abeaM be studi~ ed by WTIS (PB83- the:National Teclmual Information 5285 Part sepal Rend. - consullet AdditienP~guid~s eyb. 2329841. ~ be ~ ~1LIki.III Service, ( ebtai~froeathe is, - gwda may bs~.d fr~~ Spnia~ald.VA~*1 (788-497-498* Econom~-i*te end OECD HealthE~ Tent Gai~a~ (FR Dec. ~d1-17ø80 — ,D,velopment LOE~))Hsalth Effects for metabolism studies, as adopted by su.i.iis con _____ -