Document jmp4gn3Er0ob6yz6mYLj8852N

dTe? J, c(P* / , . / Identification end Toxlcolo-lcal Evaluation of Chlorinated Dibonzofuran and Chlorinated Uuphthalono in Two Connercial Polychlorinated dJiphenyl . J.O. Voo. Inotitute of Veterinary I'atholojy, Biltotreut 172. Utrooht, Tho uotherlandu. J.li. 1'oonen and li.L. van dor hens. lnotltuto of Volurin.ny lliartr colony end Toxicolocy, Biltotraat 172, Utrecht, Tho Uothorlandc. M.C. ton hoevor do Urauw. Ji.il. do Voo. Central lnotituto for Imtriiion and i'ood.nacoareh, 'fHO, Zoiot, Tho liothorlr.mlo. Kumiing title: Toxic inpurities lu tocinioul J'CiJ proporationa. F> >4cCc. HONS 063120 Abr.trac t In a p.rovious otudy (Too 6 iCocuan, 1370) a significant difference in toxicity wan found between 3 eo.i-.orc.lcl i'C3 prepara tions; Men mortality, lives nocrooin and chick oodona-li!io lesions occurred in 2 Ida's. In the present study it was fouml, by noons of column chroaatot-raphy and cno clirouMo^iv./uy, that in tho 23,1 diethylcthor fractions of those 2 PCJ'o polar compounds ero pro non t. A chicken onbryo assay proved the hieh toxicity of thla fraction. .Subsequent maos apaotromotrlo and Mcrocoulc- trie analyse.) rovoalcd tho identity of those compounds; a.o. totra- and pentachlorodlbonsofuriui* ?lic alaniflcauco of these lnpuritioa aro dincussod. It is concluded that ovaluntion of animaloorporlacnto with PCMs is cxtroiicl'y diffiouli due to tl.o prsoonee of chlori nated dibenzofurtuto in sous couucrdial FCJ'3. MONS 063121 Introduetlon. ' ,) . Polychlorinated biphenyls (FCjjJ havo boor, identified in tissues of nob and wildlife (jonseu, 1JC5) dollies, iiln hm & Tatton, 1967; lioldon l: Haro(ten, 19671 Koenan, Oskuup, Vocn, Brouwer, Booth, Bwart, van (lc Brook h van Genderon, 19671 Klsobi'ouj.i, Eiocho, raoicall, Iicr-nan U kirvon, 19601 Koeuan, tor. Hoover do iirnuw 4 do Voo, IjSjJ Jenson, Jolmcla, Olo.non 4 Ottoi-lind, Grid havo boon rouantly boon lupliontol iri tho mortality of oea birds in Britain (Bourns & .'lead, I96J). Coauoroiul BOB preparation's aro oxtronoly otablo and ouly slightly soluble in water. Tlioy aro ucod ao lubricants, aa heat transfer madia, in protootivo ooatinca far wood neat aotal and concxato, and in many oil.or applicatlona. Tlioy urc- compatible with paints, synthetic resins, varoishoc and vnnao, and Improve ouoh proportion of tho 00 products as their cheatcal roaiotanoo, water rosintanco, adhosivonoon and flexibility (Anonyms, 19C01 Hubbard, I364). iiowver, it has not yot Boon ostabliohod which of tho materials treated with iCj hac oontributod to tl:o present environmental contamination, Experiments with ruts and guinea pigo have shown tliat tho 1neGbtier, of TC3 uay give rlcu to liver injury (killer, 11-14). Ilydropericurdiur., occasionally accompanied by abdominal oodoma, wus found in chicks (llcCunc, Barege, i. o'Boll, 1962; Flick, O'Bcll 1 Chlldu, VjC'ji Vos d kocnan, 1970) and Japaucso (ur.il (hoennn ct ala, IJdy) al'tor ingestion of Kin of Aaericon (Aroclor) and French (Bhonoclor) oriQin, respectively, A unrkod roaoablonco between tho coo chronatoerana and does spectra In naca opeotroi .ctor analyoso Of 3 coonorcial fCB preparations (Phcnoelor BBC, Cloplicn ACO, and Aroclor 1260) was found (Koenan et el,, 1969), c The occurrence of ''thick oodonn" HUo leoiona in birds fed l'CB (Kooman ot al, 19691 lioCuno ot al., 1962) Flick ot al. 19G5) wao HONS 063122 -4i .) the roocon for a rocont conpnratlvo toxicolocioal toot with tho 3 mentioned I'C3 preparations in chickens (Von & Kocman, 1970). In thic study a significant difforenco in toxicity between tho 3 tootea compounds was found j ClopOn A60 and Phonoclor DlS chov/inc tho hl^hont. Aroclor 1260 showliy; tho lovoot portallty. Cubov.taiu.oua end abdonlual oedenu, end controlohulor liver uccroclo wan recorded Oily in thouo oliicko fed with Clopheu and Phenoclor. Hydroprricaruiu.u veu rcoordod in nearly all oliicko fed with Clophon end Flionoclor and n.v> nnmi<im.-iiv in dilolm fed with Aroclor. Uionicul porphji'i.'. v.'aa found ao a cuncral PC? effect. In tho procent study, tho ooiiuol to thnt nentlcnod r.bovo, wo doaorlbo tho ldontlfloatlon and ovaluation of oonpoundo rooponoihlo for tlio difforonco in toxicity botwaon tho 3 oonr.eroial PC? preparation:!. HCNS 063123 Fjcnorinontrl Tho PCB proparationc, whioli eontoia an average or 60, chlorlno, woro obtained fron Prodaloo, Franco (Fhonoolor Ct|), Boyer, Qomany (Clonhon AGO, Lot no. 912434) and Monsanto, U.Si.A. (Aroclor 1260, Lob no. Ali-3). The screening ncthod, used for doteotion of chickocdo.na faotor (immyssuo, i960a) was codifiod and unod to isolate from tlio PCB preparations, conpound(n) reupcnofblo for difforor.eoa in toxicity. The cloon-up stop of this procoduro van onittod. In tho colunn clirennto'rraphj 3c. actlvatod florloil (ooluim </> 10m) woo uaod inatond of 193. actlvatod aluninr-. For fractionation redijd. hexar.o wao used aa elution oolvoi t lnatoad of potrolouu otlior. Two nj PCB was dissolved in 2 nl lioxano and fractlouatod on tho florioll coition. This colunn wao olutod with lioxano until nearly ell PCB'o woro olutod (toatod by caa chronatocraphy). Tho 2nd fraction (5,' dlothylothor in hoxano) was alco tostod until no uoro FOB could bo dotcctcd. Tho final fraction (fraction 5)1 olutod with 29" dlathylothor (roatont /;rado) in hoxano, wao ovuporctod to 1 Hi. An aliquot of thlo concentrate wao lnjoctod into a 204-IB Tr.rian Asrocraph guo ohroaatocreph with elootron capturo dolootion. Tba Pyrox Glaoo colunn (9 foot x l/O inch), filled with 10>> DC 200 on Gaachrom w (80-100 Bash) was operated at 20UC with nltio,;on aa oartior caa (about 90 cil/nln.). Tho gouo prooouuro waa uaod to fraetionato allquota cf about 590 of tho PCB proporaiiono in ordo: to obtain aufflolont quor.tltioa for a chicken oubryo aouay and for ncos cpcctroaotcr analyseo. Tho PCB wao dissolved in 200 nl hoxano, The oolunn (0 32 cn) waa filled with 160 c activated florloil. The core polar conpounda were eluted in tho final (3rd} froo'.ion and, aftor evaporation, tostod by gas clirouatocxnphy. A alilokon cnbryo ascay (Vorret^ ilarliac U McLaughlin, I9C4) was used to determine the toxicity of tho 3 PC3 preparations, MOM 063124 . -0- (donoj 5,,, diocolvod and kopt a. 37C in ethanol* Croup 1 Aroclor, froup 'i Phouoclor and group 3 - Clophon) r.r.d of tho 3rd fraction of Clophon (aosoi 3rd fraction obtained from 35, 3.5, an* C.35 P-nd 0*035 ng, Clophon in groupo 4* 5 u suzi 7 rcapictivcly), Tho im.nploa, after dioDolvlng in 0.05 nl otlianol ware injected into tho air cell of frooh White Leghorn cego prior to incubi'.ticn. Vho livortof ell hatched chlcho wore removed for hiotolociccvl ccv.ninr.- tion, fixed in 10,? buffered fornalin txA procroscd in c. coi.vontlonal manner, Moreover* cryostat oootioru wero ctadiod in a fluorcoccnco nicroocopo. Hons opootreootry vao ui-cd tc identify compounds prooent in tho polar 3rd fraction. THio anclynio vao carried out uaing a goo ohronotocraph-nono spoctrc-cotor combination. For thin purpooo a C114 naos opootrocotor (Varlan iiat. Q.n.b.H*) waa coupled to a Varinn Aerograph ovon, typo A530D, with a -'yrox glaoo column (5 foot x I/O inch) filled with 10;J DC200 on Ucoehron h (U0-100 ncah), Partioular Attention vao paid to tho deoleh of tho intcrfcco betwoon tho caa ohrotiatopraph and the naoo opoctrouotcr in order to obtain tho tact operating conditions (ten Koover d flrauw U Lruunde, 1957). High rooolution naoo ocootra wore token on a double focuo opcctro- noter typo SHj (Varian Hat. C.m.b.U.). Exact naoo aonsurooonto wore done with tho peak ? notching nothod (ibasurononto voro cloo dan from eroctra which wore rocordod on photograpnic plateo prooooocd by Varian Mat# on a I.cits cotr.tarator QAMl). To obtain information about ths quantities, r:icrocoulo- ctrio enalycos cf tho 5*d fractions voro carriod out* The equip ment used vao a fohrcaim cicroeoulocotor, coupled to a 'licrutcl: i`if 220 can ohx'onato^raph, with a glaoo column (6 foot x i inch). The same 10^ DC 200 column packing v&e ucod. Tho colurji offlucatn %raro oxidised in a Dohruenn cenbaotlon furnaco and tho HCt, pru'.uccd by chlovinatod,compound, vas detootod and quantitatively uonourod nt a. T-Jcc-S o K Cc.Jl, MCNS 063125 Jlooultu 1113 rooultlQf* pan caTOiiutecxan of fraction 3, obtained with tho octal1 column (0 10ns) revealed t.io prooonco of polar compounds in Phenoclcr and Clophoa (fij.l). Tho.io vero not found in the >xoclor extraot. Tho aomo rooultc woro found with fraction 3 obtained with the large column (0 32 mm). The 3rd fraction JS, of Clophen was oubocuuontly unad In tho chicken or.'bryo acr.fl^'# Tliio assay (table l) confirmed the differences In toxicity botvocn tho 3 tooted IC proparatiou3 found in the rreviom study (Voo & Ivoomnu, 197^)* Ibrtality in croup. 1 (Arcelor) did not dlffor oignificantly (XZ tost) fron mortality in croup 9 (control). Mortality in croup 4 and 5 is dooo related; oarly mortality in tlio former# ocattcrcd uortality in tho lnttor croup* vorresj-or'Uns mortality in croup 3 (3*1? nc* Clophou) and croup 3 (3rd fraction from 3*3 &c Clopbon) indicates strongly that tho polar residua In tho letter determinoe tho toxicity of tho commercial i'Cii. jjJlncQobion of all frooh cnbryoJ ami of the ono-Uny-old ohxokn iTvahlcd no abnorunlitioe oxcopt for ono embryo of croup 3 which ohov/od.o covosre cubcutnnopuo oodena. Oodona and bool: dofouLiitioo aro diiioribed after yolk, oao iujeotion of 25 d* and 10 tie /roclor 1242, roouitinc in roopcctivoly 100/> and y-j,j embryonic mortality (llebaughlln, Unxliac, Yerrobt Mutchler *'lt*huch, 1963) d.L. otc.inoi llvor ocoiioua rovjolod no aorpholo/jic alterations. In tuo cryoutat sootion of tho llvor of the only hatched weak chicken of croup 3 a rod fluorooconco in eona lobules was con in tho u.v. uicronco^e* Tills fluorcoconco io indicative of tho porphyrin bionynthcnlo diaturbanco found in the prior otudy (Yoo U Keenan, 1970)* Haoa epoctrouotrio annlyudo revealed idoutiCvid. chlorinated oonpoiunta in fmotion 5 from Phenoolor and Clophon in cor.vr.mt with Aroolor. Tho results are <jivon in table 2. ilnou opeotra of compounds with mauo number 3^4 and 3>0, obtained fron ponlr 1 and 4 (fi^.l) oro t'ivon in fie*2 and 3* From exaot naaa noaouronento it w:.o found that the formulas of tho molecule poaho 304 and 330 voro CIc. MQNS 063126 -8- C1 jil^O^Cl^ and C^HjO^Clalio foriuulun of the fracnent Iona 241 and 275 aa-o Clj end i'rou thlo oota it oan bo concluded, that the parent ion lias a preferential fra/yaeii- tetlon for the loaa of a oincis Cl CO unit, iiaoo opectra were can; : red with the cpectiua of a compound lnolatod from toxic f.it (woolton i. Courchenc, 1?64). Subsequent otudy, uoinp X-rr.y diffrac tion, l-.no rove eled that the identity of ono of the toxic f.-.ctoro vuu 1, 2, J, 7, 0, 9-hc::aehlorouitcnzo-r-iloxln (Uoottor. L Sauxchane 1564). i'lio unoo-spectruj of thiu coiapovrd lie.3 r. olmilnr fracuc-ntatioi pattern i.o. loos of cuoccoeivo 2 maeo unlto 6; nuffjeota r. lose of 2 C1C0 unita. . ISicrocoulonetrio analyte0 of tho 3rd fraction of Ciophon and Hionoclor revealed oliroiintobrauj with tho nose ponlc pattern no in fic.1. Hie "pcrJ:" of tho totiacliiorodibonzofurun (nui:a nuuber 504) io too Mutkod by the proximity of the hoxachloronuphthcdono pooh (peak no.2) to r.nko quantitative calculations poauiblo. Pol einillor roauou peak 110,4 lu tho chronatojiran, found to bo a mixture of heptaehlorgbiphor.yl and pontachlorodlbcuaofui-an, could not be Eecxurod dire oily. Hovo7or, uhon ancumin-; that the viiolo poak 13 produced by the dlbonaofuran compound alone, a curtain maximum level cau bo lndicutod. Iliiu levol wuo found to bo 5 ppr. pontaoliloroiiltur.icoi'uran in Ciophon and 20 pin in Phtnoolor. Uouover the .nJ.nl: .urn level could bo indicated with the eutiaatod lower Halt of detection of tho unco opoctroi.iolric equipment. Undor the coi.ditione uuou thlo lavol wae 1 ppm. HONS 063121 -9- Discussion. liana opoctrouotrio re suite have not conclusively prove! the identity of the chlorinated dibontofurane. Dut cjiatbosis of thooo coaipuundu, nocooscxy for conperioon with the maos ejjoctra obtainod experimentally, van found extremely dangerous} a c.lnu'le oral dooe of 0.5-1 .0 nij/h.j trl- and tctrachlorodibousofurau caused oovore and often lethal liver neexoois In rubbito (Bauor, Jo.iuls & Spieoelber^', 1 j6l | dofuan, 193'i). Application an the oar of a rabbit resulted in ahloracno (Hauer et cl., 19GI). ihe roiatod compound 2, 3, 7, C-totrachlorodlbonso-p-dio:cln (following tho doecrlptioa of Chcaieul Abstracts), Which Day appoar in the oynthosio of 2, 4, 5-trichlorphonol by hydralyole (under praeov.ro end vith the prcoenco of sodium liydroxldo-and mathanol) of 1, 2, 4, 5-tatraohlo.vatcnzeno (Davor ot al., 1961), usually caused a lethal liver neorcslo In the rabbit after a 0Ingle oral dose of 0.C5-0.1 tc/k. Ear application, aloo In a 10 tluse lover doss oOepcxsd with chlorinated dlbonsofurnn, resulted in chloracno (Bauer ot ul., t$6l j. Linn Jor.oo .1 trlr.ck, 1952). 1,2,3,7,'2,9-i-or.achlorodibento-j>-dloxin, which con orioo in tho pyxolyoiu of 2,3,4,6-totrr.ehlorophonol (iUccfnbothrJi, Ihuai", Fireotoi'o, Vcrrett, Boss !i Ccrpbcll, 1950), 'has born loolotod end identified no ore of tho toxic sv.botanceo of the Chick Coder Factor (Vootton i: Courohone, 1364, Anonyrtuc, 1960b). Tho rclationohlp between the toxio nature cf PCD end tho Chick Codoua i'aotor has already boon dioounssd (riiok ot ul.( 1965). Air coll lsjeotion in chicken oegs of 0.05 uc hoxncortality chlox-ouitonso-p-dioxin reoultod in 100,'j osibryonic (lilccinbotlnva /' ot al., 1560). 'fiie oxparinant ulug an injootlon of tlio }rd fraotion obtained'55 mg Clophon into the air cell ronulted In 100,J nortolity (table 1). It otui bo oolculatod that the nanluun MOhS 063128 .10. dot. ,^CC do 0.2 uff pontaolilorodi, Jzofuren (uclD(j a raluo or max. 5 ppm). Thin confirms tho order of toxloity diocuoood abovo. Che combination of toxiooloeieol, pathological and ohomical analytical data calico it almost eortoin that the Compounds of maac numbers JO4 and JJO aro respectively totraond pontachlorodibonzofuron. Tho oricin of those toxio factors in Clophen AGO sad Phonoolor DP6 con poaoibly be explained by difforonooo in nanufaoturo. Particularly In tho distillation procoduro of orudo PSD, in vltioh oodlua hydroxido con bo uood (Pay & Richard^ 1947). PCD roaots with nodlunhydroxidc at olovated teaporatureo to ylold phonollo compounds, o.g. PCS is eaponifiod Into polyohlorohydroxybiphonylo by moans of oodlum hydroxido in a polyhydrlo alcohol modiurn (Anonyms, 1957). A further looo of hydroohlorlo sold oeuld produoo dibonsofuran doriyativoo. . Chlorlnatod naphthaloneo ora far loos toxio than chlorlnatod dibensofurans (Uofmonn, 1950). Choroforo the lattor may be assumed to dotormino tho toxloity of fraotlon J. Che occurrence of hydroperioardiua in ohich fod with Aroolor (Flick ot al., I9C5), nay indlcato tho proaonco of a vary small quantity toxio factor. Proa thie study It Is conoludod tliat evaluation of PCD residues in vildllfo cnimolo, based ds on sffeota and roslduos in tost animals, is extremsly difficult duo to difforonoos in toxloity botwson conmorolhl FCB preparations. Work is in progress to dotcot and Identify oliloxinatod dlbonzofuran from vildllfo apooinona. HONS 063129  L. -11nowledren Va acknowledge the contribution of Hr. S.W.A. Kaapordljl: and Hr. P.P. HI jkanp, and the tochnlool anelstonoo of l!r. J .11. 1'cnnl' This investigation was oupportod by a c*ant from tho Central Organization for Applied Solentlflo Soooaroh (T.H.O.) which mode It posolblo to aoconplieh tho anas opootra analyooo. 1 HONS 063130 12 ftaforoncc ; . ) Anonytmo, (1957) iiothod for producing plonolio oubotanooo tlirough tho traatuont of ohlorinatod dlphenylo and polyphonylo and oubotanooo obtained thoroby. Brit. Fat. 779 221, July 17 (to Frocil). Anonyzmo, (19G0). Aroolor Flastioisora. lloneanto Coopany Tooh. Pull, no. FL-J06. Anonyouo, (1960a). Changes in liethoda, 26. 011a, fata and eaxoo. J. Asa. Offlo. Anal. Chco. 51. 409-49 P. Anonyauo, (1960b). Tho chlok odosa factor, Hutr. Bov. 26. 20-30. Bauer, n., Sohula, K.11. A Splocolborg, 0. (1961). Berufllahc Vorclf- tuncen bel dor Unrstollung von Chloxphouol-Vcrblndlngtin. Arch. Gouorbopath. Oovorbohyg. 18. 530-535. Bouxno, B. & Iload, C. (1969). Seabird oleughtor. B.T.0. Howe, od. by Kolth Clark for tha BrltioU?or Ornitiiolocy, no. 36, Fay, J.V.J. & nioharda, J.l-1. (1947). Inprecncnto uaou In Coruan papor oapaeltoro. Offloa Tooh. Sorv. FB rapt. 75059, BIOS Final Bopt. 893. Fllok, B.F., 0*1)011, H.O. & Cbildo, V.A. (1965). Studios of tha olilcl: odona dlooace. 3. Slallarity of oynptoas produeod by foodlnc otilorlnatad blphonyl. Poultry Sol. 44. I460-14C5. HlClnbothaa, G.R., Huong, A., Flrootone, B., Vorret^ J., Boos, J. i Caopboll, A.B. (1968), Choalool and toxicological evaluations of Isolated cud syntketio ohlorodorivatlvos of dlbonzo-p-dioxln, Nature, 220. 702-703. Rofoann, n.Tli. (1950). nouero Crfahrungon nit hoohtoxlochon Chlorkohlenwaocorotoffon. Gxperloontello Fatbologio Phcrraokologid 812. 220-230. f MGNS 063131 Holden, A.7. l> Maroden, K. (1967). Organoehlorlno peotloidoe in ooals and porpolooo. Uaturo 216. 1274-1276. . . - -- ------- ------- - - ' \ -- I / wi*Wi.AW.lk0U hy pcorbons in British wildlife Nature, 216. 227-229. Hubbard, H.L. (1964). Chlorinated biphenyl and related compounds. Encyclopedia of Chenical Technology, ed. by Ellor Kirk, R. A Othmer, D.F., 2nd rev. ed., 209-297. Jensen, 3. (1966). Report of a new chemical hazard. Hew Sci. 32, 612. Jenson, S. Johnels, A.G., Oleaon, II. A Otterlind, 0. (1969). DDT and PCD in narine animals from Bwodlsh waters, nature, 224, 247-250. Koemon, J.1I., Oskamp, A.A.G., Veen, J., Brouwer, S., Rooth, J., Zwort, P., v.d. Brook, E. A von Genderen, H,,(l967). Inseetioldes as a factor in the mortality of the sandwich tom (Sterna sandvlconsia. IIodedpRijksfacvLandbouwwetonaohappen Cent, 92, 641-054. Xoeman, J.R., ten Soever do Brauw, II.0. A de Too, H.n. (1969). Chlorinated blphonyls in fish, mussels and birds from the river Rhine and tho lletherlando coastal area. Nature, 221. 1126-1128. ' Linn Joneo, E. A Krizak, II. (19C2). A technio for tosting aensgenio potency in rabbits, applied to the potent acnogen, 2,5,7,8,- tetraohlorodibenzo-p-diozin. J. Invast. Dormatol. Six. 511-517. IloCuno, E.D., Savage, J.E. A O'Dell, B.L. (I962). Hydropcrlcardium and ascites in chicks fad a chlorinatod hydrooarbon. Poultry Sol. iJj. 295-299. MoLaughlin, J., Marllac, J., Torre H.J., Ilutohlor, II.K. A Fitzhugh, 0.0., (1965). The injection of chemicals into the yolk sao of fertllo eggs prior to incubation as a toxiolty test. Toxicol. Appl. Pharmacol., 760-771* Miller, J.W, (1944). Pathologio changes in animals exposed to a oommoreial ehlorinated diphenyl. Pub. Health Rep., 59. 1005-1093. MOMS 063132 - i) . 14- ) Kioobroueb, H.U., Rloche, P., Poaknll, B.3., Horaon, O.C. & Klrvon, ii.B, (1966). Polyohlorlnatod blphcnylo lu tbo global ocooyotaa. tlaturo, 220, 1093-1102. Con Hoover da Brnuv, II.C, & Srunndo, C.Z. (1967). Ircobninoo nit elner optinicrtcn Koppldng von Dus ChrooatoGraph uud llaooen- apokl.ruL.ater. Zeltochr. f. .Inal. Chon. 229. 321-333. Vorrott, M.J., Ilarliao, J. & iicLaughlin, J. (1964). Vao of tlio chiok.cn onbryo in tho aooay of aflatozln toxielty. 4. A93. Offio. Acr. Chon. 21003,-1006, Too, J.Q. & Koonon, J.U. (1970). Conparatlvo toxicoloclocl study with polyclilorinatod biphenyls In obickono wltli apodal roforcnce to porphyria, odena foxuatlon, livor noieroaln, and tloouo roaiduoo, Toricol, Appl. Fhamacol. In prooo. Vootton, J.C. & Courobono, V.I>. (1964), A contribution to tlic knoulodco of tbo otruoturo of two hydroporiaardiun-produoina fnotoro from a tuxio fat. Apr. Food. Chou. 12.-94-93. MQAIS 063133 Table 1. Chicken Etabryo Assay of Chlorinated Diphenyls Aroclor 1260, Phenoclor DP 6 and Clophsn A60 and the 25 per cent Diethylcther Traction of Clophen A60. Group Corpound Dose/oeb& Rusher Rusher of Per cent Hatoh Infortile Eggs of Fertile Eggs Reaarba 1 Aroclor 1260 3.5 ne 2 Pbenocler DP 6 3.5 Be 5 Clophen A 60 3.5 oc 4 Clophen A 60 3rd fraction of 5? mg 5 Clophen A 60 " 3.5 ne 6 Clophen A 60 " 0.35 oe 7 Clophen A 66 " 0.055n6 8 Ethanol ' 9 Control , 15 15 20 15 15 15 15 ^20 20 0 0 1 0 0 2 2 2 0 00 0 5 0 .7 92 100 94 90 Kost enbryoe diod durin~fln half of inoubatlon period. Kost onbryoB diod during fin half of incubation period. All cnbryoo diod during firs four day3 of Incubation porii Host eabryoo died dorlng fin half of Inoubation poriod. aThe sanplea, dlesolTed in 0.05 nl ethanol, were Injeoted into the airooll of frech eggs before Incubation. MOMS 063134 Table 2. Retentions and Collected Maos Speetronetrlo Data of the Peaks in fig.1. Peak no. Relative retention ' Kass nunbcr(s)'>, and mmher of chlorlnat&toDs/aoleeale Identity of the eompotmd(s). 1 1.40 2 1.53 5 1.74 4 2.42 5 3.48 304(4CX) 332(601) 358(601) 392(701) 358(501) 392(701) 566(701) tetxachlorodibenzofuxan hazachloxonapkthalene hexachlorobiphenyl hoptachlorobiphenyl pentaohlorodibenaofnraa heptaohlorobiphenyl heptachloronaphthalene detention related to dieldrin 1 ''Haas motor calculations bgsed on Cl 55 MCNS 0 6 3 1 3 5