Document 717kzEexL2MRr9kxK9enL3QEB

Monsanto (j ') PNOM (NAMt IOCOIONi R. E. Keller - RID Laboratories - South Second Street June 30, 1970 CHLORINATED DIBENZOFURANS - PCB ______ ______ cc H. S. Bergen - GO W. R. Richard - GO E. P. Wheeler - GO M. w. Dietrich - Res. 2 E. M. Emery - Res. 2 TO : W. B. Papageorge General Offices The attached letter and manuscript have been received from Dr. J. G. Vos at the University of Utrecht. It is in reply to our request for details of their work on the identifica tion of chlorinated dibenzofurans in PCB. Please note that this manuscript has been sent on a confidential basis and it is not for general distribution. We will attempt to provide chlorinated naphthalenes requested for gas chromatography work by Vos. Bill Richard thinks we might be able to obtain these commercially .J' Dr. Emery is asked by copy of this memo to determine if we have any of these compounds in SRC. Drs. Dietrich and Emery are asked by copy of this memo to please review the manuscript and determine the difficulty of applying this technique to our Aroclor products. Assuming this is within our capabilities with a reasonable amount of effort, we will look at Aroclor products used at Bio-Test for feeding studies and compare results with those for current production grade materials. db Attachments fcR. E. Keller "Vv\yc>-*v<rc t /u7 (jW7 teJ/^ l 0s* I o T/to I (/A(y ? '/V % 6 rrvK MUMS 099146 INSTITUUT VOOR VE1ERINAIRE FARMACOLOGIE EN lOXICOLOGIE DER RIJKSUNIVERSITEIT TE UTRECHT DlkECUUR: Piol. H. VAN GENDEREN No Bilchl op: BeOeflende: Bljlnoeo: + 0 UTRECHT, June 24, 1970. 172 leletoon No. 715544 Dr. R.E. Keller, Applied Seiences Section Monsanto Company, 1700 South Seoond Street, ST. LOUIS, Missouri 63177* USA Dear Dr. Keller, Enolosed you will find a photocopy of the manuscript oonoeming the identification of chlorinated dibensofurans in PCB. use fractionation, QC/mass spectrometry techniques* the details are given in the manuscript which is not for general distribution sinoe it is not yet accepted by the Editors of the Journal of Food and Cosmetics Toxicology. During the visit of you and your oolleagues at our Institute ve mentioned a.o. severe skin lesions (hyperkeratosis) in oattle. The floor of the stable in which the animals were housed was ocrvered with Booond-hand conveyor belts, covered with a new anti-slip layer. Caa chro matographic analyses of a sample of the rubber mat and of the liver of a severely affeoted oow revealed a peak pattern whioh was very indicative for PCB, as we also mentioned during your visit. Moss speotrometrio ana lyses of the rubber revealed compounds with molecular weights of probably penta- and hexaohloronaphthalene, well known for their hyperkeratotio aotion in oattle. . Chlorinated naphthalenes are not for sale and sinoe they are very important for ue as referenoe compounds in gas chromatography, we would like to ask you to mediate for the eupply of ohlorinated naphthalene mixtures and perhaps a pure sample of oota ohloronaphtalene (perchloronaphthalene) . Dr. Koeman told me that he will send you a total-body sample of a oormorant containing a high oonoentration of PCB's, for a comparison of the quantitative aspeots of the analytioal methods. Sincerely yours, Institute of Veterinary Pharmaoology and Toxioology, University of Utreoht, Biltstraat 172, UTRECHT, fhe Netherlands J.Q. Vos. MONS 099147 f ^0> * * .A ) Identification and Toxiowlo~ic*kl Lvnlujlion of Chlorinated Dibonsofuran and Chlorinated Uuphthalono In Two Conaerolul lolyuhlorinnted nlphcnylo* J.C. voo. J.notituto of Voterinary I'otholo^y. UlltOtrt.it 1T2, Utrecht, The othc-rl ;uuiu. J.!l. Koeman and U.L. van dor l t.no. lnotiVoto of VotuxlnMy Vhariv eo) rntl Toxiculory* Dillotraut 17*, Utroclit* Tho Kotherltndc. H.C. ten Uoevor do Urauw. Ji.lt, do Vos. Control lnotituts for liutrltl-on and rood-flotoarcJi, THO, Zolot, Tho Nothorlrjids* iiuiuilnc tltlet Toxic inpuritiou In toohiloul iCd propc-ratlono. )% C\ _ ' *< A'1, ^ -* zrvrt HONS 09914a o Abr.tr;.et> In a proviouo study (Voo u Kooum. v,'7w) u i.utficu.ut difference In toxicity was found botuoeu 3 coa-crciLl iz-t ] ] tlons; lilgli Mortality, livor nocrooio and chick oodoiaa-lika leoionu oocurrcd in 2 iCii'y. In the pi-oocjit utuuy It w.ia found, by veunu of colum chrouutO{rraphy u:d (jay ohrouuto(;rvhy that in tlio lUotltyi* Ot):ot fruotiono of thoao 2 Wil'u polar eoui'omulu o.ro yveuent. U oliicUen embryo aooay proved the hieh toxicity of till a fraclioru Jubsoquont tiaas rpootruuutrlo and .lcrocoultv trio tutalyrieo rovoaled th identity of thoi< cout)/ouidoi .o iotra- end yenta- oblorodlbontofurun* ?bo ai^-nificanco of thcoc lupurltlas ore dioouocod. It la concludod that ov'duntion of anlcnalooxporliuonto with PCtt'a lo oxtreiicliy difficult due to tlio proaonoa of chlurl- notod dlbvnaofurtuio in ootio coiauierdiol rCJ'o* MONS 099149 Introd-iotlo.i l'olyohlorinatad biphonylo (iCtf) have boon Identified la tissues of fish and wildlifo (janooi., 1^CC| iloluos* biii-ionj & Tatton* 19671 lloldon l> Narodon* ly67| &oenan, uofcnnp* Vcon* Brouwer, booth, Zvurt, raa do Brook 4 van Conderon, 19671 K100trough* Kiocho, rookall, Herman 4 hirvon. 19601 Koomnn, ton Noovor do Brnuw U do Voo* ijiyi Jonoon, Johnnie* Olonou 4 Ott .'illnd, and have boon ryuuntly l>oaa luplloatod iti the nortollty of eoa blrdo In Britain (lsourno 4 Head, 1969), Comuorolul fdB propr.rutloao aro oxtrouoly atublo and only silently eolubl# la wator. Thoy aro uood ao lubricanto, aa boat tranofor neilia* in protootlvo ooxtinco for wood *0** uotul and coacroto, end In o&ny othor Bpilioutlo-m.. Tnoy arc compi-tibl* with paints* synthetic rosins* varalshso and waxoo, and iajrove ouoh proportioo of t)eo produoto ac their choaioal raaiotancu, wator roalctanee* odhoalvonoee and flexibility ^Anonyuua* I96O1 Hubbard* 1964). Uo&or* it luia not yot boon cstubllohod whioh of tha iuaturlolo tioatod with iCb has ooutvibulod to thu proocnl environmental contamination. intpoxluonte with ruto and guinea {leu have shown tlmt tl.o liitoutlbn of fCJ uay givo rloo to liver Injury (tlillor, 1^44) HydroporicjuJiun, wccacioiUillj accompanied by abdouian-X oedema, was found In oI.IcaQ (licCuno, Lavaca L o'BaXl* lybiij llior., u'ball 4 Childu* VyC|^t Voo 4 Loouttix, Vj70) and Jupunteo qui.il (kosmtin ct al** 1969) aftor ingootion of XCii of leorloan (xroclor) and Frouulr (ibonoolor) origin* respootivoly. k marked roooL.blunoo between tho goa efiroijutonruBa and ciacs opoctra in naco opootxonotor analyaeo of 5 oonuorcial PCB propurutlono (Hivnoclor Dr6, Clophon A60, and Aroolor 1260) wau found (Koowan at d. 1969)* Tha occurrence of "Chiok oodoua" like lesions in birds fed PCI) (Koanun ot el. 1?69| MoCura at al.* 1962| Fliok at al. 1965) woo ~V,T*Ti ~ - "4d'a*i MUNS 099150 6 4- 3 tho ror.con for a rocent cooptirativo toxj col&rical toot with tho 3 ncntlonod I'CJ) preparations in ehlOkono (Voo & Kociooo, In thlo otudjr a l^nifleont dlfforenoo in toxioity botwecn tbo 5 tuotou no.ipounda via found) Clojfon />60 and Hwnoolor DIG allowing tho liichoot* Arcelor 1260 shooing tho lovoat Dorttlity, C-ubevtajicouo end ubdoutual oedcua, uul ocntrvlobular liver nccrocio vno recorded oiily in fchouo oiiicUa fed with Clophcit mid Hicnccler* HydropcrJ enrdiun veu rooordod in noarly all chic).a fed with Clophon ri.cl Fhunoclor end oily oucnnlonelly in ohloJ-.n fod with Aroclor. Chemical porpliyrl;: wiuj found ud u general l'CU effeot. In the prcoont otudy, tho oouuol to that uonUenod c.bovo, vo doaoclbo tho identification and evaluation of oosnounAo rooponolblo for tho dlfforonoo In toxioity bntwoan tho 5 oomno^olai FOB proparatlono* HONS 099151 ) 'W>r5.M0iit."^ -2- O Tho PCB proparationo, whioh contain on nyorarjo of ' 6 60,i oblorine, wore btainod frora I'rodoloo, franco (Fhonoolor iu(), Ikvyor, Oorreany (Clophon ACO, Lot no* 912454)* And llonocnto* U.S.A. (Aroolor 1260* Lot no* Ak-3) The screening method* wood for detection of oiilc'(ocdna faotor (fcnnnywuo* 19^0n) woo ttodlfiod and uood to isolato fron the PCD p.Toparutlonnf conpound(n) roopono.lblo for dlfforencou in toxicity* Tho oloaii-up stop af this procedure was onittod. In tho oolunn elirematorraphl 0fi. aotlvatod florloil (ooluj.in 0 10u.) wao uood inotoad of Vj"* activated aluninr.* l`or fractionation rodljd. hexane was uood eo elution oolvont lnotoad of potrolouu other* Two PCD wao dloeolvcd In 2 al hcaano and fructionatod on tlto florloil ooluan* Thlo colunn was olutod witn hoxano until noarly ell PCV'e wore elutod (teotod by c*o chromatography). The 2nd fraotlon (5, dlotlvylothcr In hoxano) was also tested until no uoro PUD could bo dotoctod* The final fraction (fraction 3) elutod with zyA diothylothor (reagent grade) in horuno* was ovuporatod to 1 ul An aliquot of thlo oouoentrute mao injoetod Into u 20<l1b Varinn Aerograph gas ohrooategreph with elootroa oapturo delootlun* The Fyrox close coluun (3 font x I/O lnah), fillod with \o-A DC 200 on OnoonroA u (80-100 noth) wao oporutod nt 20U0 with nitxo.ron ea oartier fao (about 'jO ml/aliu). Tho souu psooodure van uood to fractionate aliquots of about an of tho l'CD preparations! In order to obtain eufflolont quantltloo for a ohlckcn ctihryo eoouy and for naoe opoctronotcr analyooo. Tho ICd was dlooolvod In 200 ul hexruio. The coltu.-Ji (0 32 co) wao filled witli 100 c notlvatcd florloil* Tho more polar conpoundo woro eluted In tho final (3rd) frootlon and, after evaporation* tootod by c^a olirouutocrnphy* A ohloUan eubryu assay (Vorrstt* Hurll&t k )usL&uglilin, 19C4) was uood to determine the tojclolty of tho 3 PCB prvporutlono* MONS 099152 1V (donor 55 uioaolvod and kopt at J'/C in ethanol* Croup 1 - Aroclor, trtnxp Ihonoolor and ({coup 5 Clophon) tnrt of the 3rd frnotlon of Olophen (uoeot >rd fraction obtained from )'jt 5,71, an* O.55 and 0*033 nc Clophon in (,-roupo 4 C a:?i 7 renpc-ctivoly)* Tho uumplos* after diaoolvlnc In O.Oli ol ethanol wore Injected Into the air coll of frooh While Leghorn cuxo prior to lnrnbutlon. Vho llvorwof ej.1 hutched chtcko wore roiuovud for hletolof;lonl o/cnlna- tlonv fixe* ih 10,j buffered fornolln ir.d procaooed In r co\o'itJonal laannor* Horootor, oryostul eootlone woro otadiad In a fluorouocuce nloroocopo. Hans rpootronotxy vao uood to Identify eonpotuulo piooont In tho polar Jrd fractious* Thlo anrlyalo wac oarrled out uolng a Co oliroaato(jraph-nuoo spsetnuotor oonbiurvtion* Vor thin purpose a 014 nnso epootroootor (Vnrlan Jlat. U.n.li.li.) was ooupled to a Varlan Aoroffrnph oven, type AJ.SOO, with a /yrox glaoe coluat (5 foet x l/8 Inch) filled with 10;1 DCL'DO on Uunoliroo 4 (CO-100 uoeh), Pnrtioulor Attention vao paid to the dcolffb of the intcrfcco between tho fras ohroMotograph and tho naoo opoctroyotcr In order to obtain tho boet oporatlna conditions (ton ioovor de JJrauw & LrunntJo, 19^7) UIqVi rooblution ejaoo opootrs wore taken on o doublo focuo opcotro- soter type (Yarian Mat* O.o.b.H.), Hsnot naoo nonsurouonte wore dona with tho peak j nebohinc nothod (Jlaaouronenta wore eloo dona from spectra whloh ware reoorded on photoffraprtio plateo prooeonod by Tarion Hat. on a 1-eltz comparator OA.'ll). To obtain Information about the quontltios, nlorocoulo* octrie onalyooe of tho Jrd frsotlono wsro cnrrlod oat* l ho oqulp- nnnt uetd wso a Dolirsans aicroooulocotjri eouplod to a .'Ucrotok li'f 220 c0 ehromatofrapb, with n nioae oolu-nn (6 feet x { lnoh)* The wr,o 10/ PC 200 column paoklna v&e uoed. The ealurai effluente ware cxldlied in a Dohnaann combustion furnace and tho )iCl* pro-'uend by ohlorlnated eompounda* van detected and quantitatively meaaurod in 0 T-JOO-D titration coll. 7* Th* roouiting gan riiroijotocxtun of fraction J, octuinM wit)) the emll oolum (jf lOno) rovoulod Die proeonco of polar corapoundns lii llitenoclcr end Clop)mn (fi.j.1). Those were not found in tl.o Aroelor oxtraot. 7ho tmao rosultc vero found with fraction ) obtained with the large column (fif 3? ran). The 3rd fraction it cshiaJtted of Clophen w.io oubocauontly unod In tho c!ilo'<o:i o:iliyo array. T'nto nomy (table 1) confirmed Die dltferonooo in testoity between the 5 tooted PC* proporatious found in the previous otudy (Voo i koanuitt* 197*0* -iortallty lr. giof. 1 (Ai^clcr) did not differ significantly (X*- tost) fron mortality in group 9 (oontrol). Mortality in cron? 4 end 5 is dooa related* ourly nortaiity in tli former* oenttored uortality in tho latter grout* Coircojondinj mortality In group 3 (j.> ng Clophou) and croup 3 (3rd iruotiou from 3*3 (? Clophon) Indioctou otronyly that tho polar reelduo in the lrttor deterninec tho toxiolty of tho commercial PCH l>incootion of all fro eh enbryoiand of the ouo-day*old ohloks rrvo&lcd no abnoruolltlea except for ono orabryo of (Troup 3 which ohoi/cd.0 oovore cuboutrnopuo ooderaa. Oodona and Loo): doJoiuiltloa are dinorilwd after yolk nao iuJooUon of 23 da and 10 rag /ruclor 1?42, resulting in roopcctlvoly 100,> and y;},) ombryoulo mortality (iloLauflilin, llarliac. Verrott Mutoblor a Pltaliuch* 1963) d.u otnlnod llvor oeotlour rovjoled no sorpnologic ultomtione* In tm oryoutat oootion of tho llvor 0/ tho only hatched woo): uiiiaken of group 3. a rod fluorocioonee in eorao lobules vao ocm In l)m u.v. uicronoojio. This fluorosoonco to indleetlvo of tho porphyrin blonynthooin disturbance found in the prior otudy (Voo U Kecrnn, 1970). ltaon epontronotrio analysis rovcnlod identic.Q ehlorinntod oo^pounAo In fxaotion 3 fron Mtenoolor urul Clophon in oontraot with Aroelor. The rooulta ore givon in tablo 2. Jlaoo ejieotra of coopoundg with aauo uumhor 304 and 330 obtained froa peak 1 and 4 (flg.l) are glvon In fig*2 and 3* From exaot ansa moaoureoento it was found that the formula* of tho molecule pool:* 504 and 390 worn MONS 099154 1 -- t c .0- 1? 4 and C.JUO^Cl,-. 'llio forwulua of the fx\.,;t.ent Joan 4 12 ? J 241 and 27'j exo C^U.^CIj and, Prou thin beta It can Vs concluded that the parent Ion Iiul a proforMitial iia ..an- te.tlon lor th*i lour nf o ninple C1C0 uviit. iiono npeetr;: were e worst'd with tJis opoctiun of u compound laolcitod from toxin fat (r.uoiton i. rouichcnc, 1J.'64) Cuheoruent ov4y, unlrf X-rr.y iMff ai- titin, Iro revr* tied that. tho JdojAJ.tv of one of the toi;Jc f.rtora vau 1, ?, >, 7 t't 9':e"u.chlorodU.cnr-i-.UoxJn (i.'octton <- vt-uvehons, 15>64)* `fhc waoo-sptctru.3 of thlu eomvow.i 1k-.o (. oimilnr fnv .dilation ruttern lo looo oi tiuoconulvo 2 toaao units 6} ougjcoto n loaj of 2 C1CU uuita. I.iorocoulos-ttrio a-uulyueo of the Jixi fraction of (Jlophoa and Hionoclor xcvoalad oljroi.ilo^riMUj with Uu tmjua pool; potti'in ou in The "pcij:" of tho tctruchloi'oddbom'ofurau (e.ui;a dauber 7*04) io too uuokod by tho proximity of the hoxactilorgjinpiithhlu/io pool; (peal; no.2) to nl:o quantitative nulculatiomj puuotUlQ* tui airtllor roruon peak no.4 iu the cluroiuto:.rui>, found to bo n ulxturo of heptuchlorybiphenyl 'and pcntuch3orodtbcnv.ofu 'an, could not to rxucavou dlrootly. )ioco7cr, when uojuuirr; that the wholo pool, is yroduooil by the dibom.ofurun compound tlens* u detain nu>;ii.un iovr.l cou bo Indicated. Uiiu Isvul wuu found to bo b Pi' poitao!iloroa4bu;co.'UiiUi In ClupJitn ami ::b pjw in Jlitnoclor. Uouuvci tho .alnluuu ltv.il could, bo Indicated with the lotlLUlcd luwor Unit of detection of tho iun/m upoetronotri <n'ipnont, Under the. condi tions uuod this lsvol wuo 1 ppm. HONS 099155 1 * Dt.-c-c-.loi'. liiiuo opoetrootrio resulte have not eonalunively proved the identity of the chlorlnntod lUbeneofuruua* But cymtho.ils of those ooupounda* noceosury lor coupcrioon vith the maoo ;>octro obtained experimentally* vuo found extronely dan^trouo) a oin,,le oral does of 0*5*1 *0 tri- and totraclilorodibonzufurou cauued soves'c and often lotbul liver noerooio in rubt-ito (bailor, Je mis & Spiegel ber^;, 1?6l| hofnan, /ip.'lioation on t.i* oar of a rabbit moulted in ahlorscno (Uauor et ai. 1961). lhe relatod eonpound 2* )t 7$ C-tetrachlorodibomo-p-dioxln (following tho doeeriptlon of Cbouicul Abotraoto)# iftlcb nay appear in t>t* ejrn* theoie of 2f 4t 5-trlohiorphenol by bydrolyoie (under preaoure aad vith tho prooenee of oodlun liydroxido -and nothanol) of 1, 2, 4, 5*te traohlorobenoene (Bauer et alM 196I) uoually ocuood a lethal liver ueoroeie In the rabbit after a oinclo oral dooe of 0*05*0*1 uc/kc* Cor application, also In a 10 tlrioe lower dooo oocpcxod villi chlorinated dlbon&ofuron, reyul>od In chlorycne (Bauer et ul., 15611 Linn Jonoo & hrlcck, 156?)* 1 2*37*u,9-ior.a- chiorodibcneo*j>-dioxiu which cun arlao in tiio pyrolyeiu of ^*5,4C-tiitrcvo))loroyhonol (Ultclnbothiui, lluujic, lirootono, Vn#tt, Uooo 1 Cnrpbell, 1p60), Tuio born ioolototl and identified ce opo of tho toxic Bubotancoa of the Chick Ooueua factor (lootton L Courohene* 1964 Anonynue* 1p6Qb)* The rclutiouuhlp botveen tho toxio nature of PCD and tho Chlok Cedoua laotor hoc already boon dieounuad (flioli at ul** 1963)* i-lr coll injeofcion in chlcioa ogcu of 0*05 ug htxa* mortality ohlorouitonfcoy*dioxln reoultod In 100, oabryonlc (Ulcclnbothcua at ul*t 196b)* The exporinont using an injootlon of the Jrd h- 100,i wx'tality (table 1) It oan be oaloulatod that the i-mxluuu HUNS 099156 I t. i I 3 L.. doot/occ io 0*2 us peutaohiorodlbonzofuron (using & toIuo of aax. 5 ppn). This oonflros tho or&or of toxiolty diocusood *bavo Tho oonblnatlon of toxlooloclealv patltoloffloal and ohoaloiX analytical data nckoo It aluoet oortain that tho coapoundo of naoo nuubcro J04 and 550 arc roopootively totraand pontaolilorodlboncofuran. Tho origin of thooo toxlo factor* in Clophen A60 and Phonoolor DP6 con pooolbly bo explained by difforoneoo in nanufaeturo. fartloularly In tho distillation proooduro of orude PCi)t In whlob oodlum hydroxide oan bo uood (fay & IlleharA, 1747)* MB roaoto with sodluahydroxldo at lovatcd tsoporaturoo to ylold phonollo ooupoundo, e.g. I'Cli Is aponlflod into polyohlorohydroxyblphanyla by eeano pf sodlun hydroxide In * polyhydrlo alcohol aodiua (Anonysuo, 19J7) A further loos of hydroohlorio acid oould produce dlboanofuran derivatives. . Chlorinated naphthalenes are far looo loxlo than oJilorlnatod dlbontofurano (Hofmann, 195) Therefore tlie l&ttor oay be aaouBCd to dotorulno tho toxiolty of fraotlon 5* Tho ooourronco of hydroporloardlun In ohlok fod with Aroolor (Fllok ot al. 19<S5), nay indloato the prosonoo of a rory email quantify toxlo factor. Proa this study it lo oonoludod tlmt evaluation of PCD rosiduoe in wildlife anlaale, booed do on offooto and roelduoo in toot anlmalo, io extremely dlffloult duo to dlfferonooa In toxlolty botveon ooanorolii PCB preparations, Vork is in progress to doteot and identify chlorinated dlbensofursn froa wildlife sposinoao. MONS 099157 Aeknowlod/rcaentB* Wo acknowledge the contribution of Hr B.W.A, Kauperdljk and lir* P.P. Hljkmap, and the toohnioal aoalstanoo of llr Jn ronnln. This investigation wao supported by a grant froa t)o Central Organisation for Applied Oolentlflo Rosoaroh (T.If.O.) which node It possible to eeeonpllsh the aaae epeotro analyaos* MUNS 099158 L-UA .J.j. Jfc, Heforoneo** Anofiyoua* (1957)* Method for producing p>euolio eubotanoo* through tho troatuont of ohlorinatod dlphenylo and polyphonyio and Bubotanoao obtainod thereby* Brit* Pat* 779* 271* July 17 (to Frogi!)* Anonyauo* (l?60). Aroolor riaatlolaora. Iloneaato Company loeh. Bull. &o fl-jqg* Anonyauo* (l?(SOo)* Change* In ItothodO, 26. 011a* fata and waxoo. J Ao* Offlo* Anal* Chon* 51* 40?-49t* Anonyauo* (l<?6tib)* Tho oliiok edooa faotor* Hutx* Hov* ZC, 20-JO. Bauer* 11.* Sehule* K.H. A Splogolbozg* 0. (Iy6l). Darufllohe Vorclf- tuncea bal dor Ueratollung ran Chlorphnnol-Vorblndlncon* Aroh* Govorbopatb* Oovorbohyc* 550-555. Bourne* B* A iload* C* (1969)* Seabird eloughter* B.T.O, Nave* ad* Fay* J.W*J. A lUohorde, J.M. (1947)* Imprecnont* uaad In Oursian p&por opnoitor*. Offlo* Tooh. Serv* P& rapt. 75059* D100 Final Rapt* 695* Flick, D.F.* O'Doll* R.O* A Chlldo, V.A. (1969). Studio* or tha ohlok edeoa dloeose* J. Siuilarlty of aynptoas produood by focdlac ohlorinoted blphonyl* ivuitry Sol* 44. 1460-1465. Hlgglnbothaa* C.R.* Huong* A** Plzoetona* D.* Yerrett* J* lt*oe* J* A Caapbell* A.B* (19C8)* Ohealoal and toalooleclool evaluations of Isolated and eynthetlo ohloroderlvatlvo* of dlbenao-p-dloxln* Batura, 220* 702-70J. Rofoonn* Il*?h* (1950)* Houero Lrffthrungon nit boohtoxlechon Chlorkohlenvaaeexetoffen* Ksperlaontell* Fathologl* Vhernakologle, 232. 220-2JO. Holden* A*T* A Marodto* X* (1967)* Organoelilorint p**tloldea 1a ooal* and porpolseo. Hature 216* 1274-1276* V HONS 099159 -13Bolmes, S.Q Simmons, J.1I. 4 Tatton, J.v 3* (1967). Chlorinated hydrooarbono In British wildlife. Nature, 21 Ct 227-229. Hubbard, H.L. (1964). Chloriaatod biphenyl and related compounds. Encyolcpodia of Chemieal Technology, od by Bllor Kirk, R. 4 Othmer, D.F., 2nd rev. ed., ^ 209-297. Jenoen, 3. (1966), Report of a new oheaioal hatard. Ilew Sol. 3?. 612, Jenson, S. Johnels, 1.0., Olaoon, M. 4 Otterlind, 0. (1969). 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Toxicol, Appl. Fharuaool. In pzoao. Wootton, J.C. & Courohono, W.L. (1964). A contribution to the knovlodco of tho struaturo of two hydroporioardiun-produolng fnotoro from s toxio fat. Apr. Food. Chou. 12.-94-90. MONS 099161 Table 1. Chicken febryo Assay of Chlorinated Biphenyl* Aroclor 1260, Pkenoelor DP 6 and Clopben A60 and the 25 per east Diethylcttsr Fraction of Clophcn A&0* Group Compound Dose/egg* Buaber Kusber of Per cent Batoh Infertile Eggs of Fertile Eggs Borarks 1 Aroclor 1260 5.5 B* 2 Phenoclor S? 6 3*5 ng 3 Clopben A 60 3.5 t 4 Clopben A 60 Jrd fmotion of 55 eg 5 Clopben A 60 * 3*3 ng 6 Clopben A 60 " 0.35 ng 7 Clopben A 60 6 Ethanol ' " 0.055ag 9 Control 15 15 20 15 13 15 15 20 20 0 0 1 0 0 2 2 2 0 60 0 5 0 7 92 10C 94 90 Koot embryos died durln^^rst half of Incubation pc rid ' Meet eobryoe died during first half of Incubation period* All eabrjos died during first four daps of Incubation ported. Rost enbryos died daring first half of incubation porlod* * MONS 0 9 9 1 6 2 *Th sanplea, dissolved In 0.05 al ethanol, vere Injected Into the alrcoll of frech eg-^B before Incubation* / / fi f j` k s lj i' i *o Table 2. Retentions and Collected Mass Speciroaetric Data or the Peaks in fig.1. Peak no. Relative retention Mass mmter(s)^, and number or chlorina'v&iens/solecule Identity of the conpatmi(s). 1 1.40 2 1,53 3 1.74 4 2.42 5 3.49 - 304(401) 332(601) 358(601) 392(701) 338(501) 392(701) 366(701) tetrschlorodibenzofuran hexachloronaphthalene hcxachlorobiphenyl hoptachloro biphenyl pentachlorodibenzoruran heptachlorobiphenyl heptachloronaphthalene detention related to dieldrln 1 ^Hass number calculations bsed on Cl 55