Document ba43drr6Gy29jJYgL7Q01rBvO

FISHERIES RESEARCH BOARD OF CANADA Technical Reports FRB Technical Reports are research documents that are of sufficient importance to be preserved, but which'for some reason arc not appropriate for primary scientific publication. No restriction is placed on subject matter and the series should reflect the broad research interests of FRB. These Reports can be cited in publications, but care should be taken to indicate their manuscript status. Some of the material in these Reports will eventually appear in the primary scientific literature. Inquiries concerning any particular Report should be directed to the issuing FRB establishment which is indicated on the title page. FISHERIES RESEARCH BOARD OF CANADA TECHNICAL REPORT NO. 272 I PCB AND OTHER INDUSTRIAL HALOGENATED HYDROCARBONS IN THE ENVIRONMENT BY V. ZJLtko and P.M.K. Choi This Is the forty-third FRB Technical Report from the Fisheries Research Board of Canada, Biological Station, St. Andrews, N. B. 1971 NEV 036107 **U n x ^ . - - i. ABSTRACT - Industrial applications of polychlorinated biphenyls (PCB), chlorinated naphthalenes, chlorinated paraffins, chlorinated benzenes,.polybroraiinated biphenyls, chloro-bromo biphenyls, chloro-hydroxy biphenyls, and halogenated carboxylic acids.are described. A list of other halogenated hydrocarbons and derivatives, used as solvents, fumigants, refrigerants, flame retardants, aerosol propellants, heat-transfer media, hydraulic fluids, etc., is Included. Current methods J for the measurement of PCB are reviewed. PCB and p,p'-DDE levels in fishes and aquatic birds, reported in the literature, are summarized. Recently determined PCB concentrations'in Atlantic salmon, bluefin, yellowfin, and skipjack tuna, bluefish, swordfish, and blue marlin, cod, white hako, plaice, and redflsh, double-crested cormorant, herring gull, black duck, and guillemot are presented. The determination of polychlorinated ter phenyls (PCT) ih"biological 'samples' and: the -- behaviour" of p'olybroraina'ted biphenyls and .polychlorinated dibenzo-p-dioxins and-dibenzofurahs*during cleanup and ga s_ chr oniatogra phy..1s_r.desc*ribed r ''Biological *effec ts of ' PCB and other halogenated hydrocarbons are reviewed. The sources of PCB leakage into the environment and their circulation are discussed. Analytical procedure-for the determination of PCB and organochlorine pesticides based on the modified cleanup chromatography of Holden and Marsden is described. NEV 036108 ^ ^ -^- _ .4 fI INTRODUCTION The persistence of chlorinated hydrocarbon pesticides in the environment `and their accumulation in the food chains is well known. A variety of halogenated hydrocarbons is used In industry and, as demonstrated in the case of PCB, the environmental behaviour of these compounds may be similar to that of chlorinated hydrocarbon pesticides. Several PCB review papers appeared recently in the literature (Risebrough et al-., 1968, Risebrough, 1969, Veith et al., 1970, Gustafson, 1970, Peakall et al., 1970, and Reynolds, 1971) Interest in environmental aspects of PCB is growing fast and.much new Information became available. In the following, only articles not cited in the above reviews will be mentioned. Other halogenated hydrocarbons of industrial importance are also included in the literature survey. Uses of PCB and other halogenated hydrocarbons No data on the production or consumption of PCB in the world are available except fo*r Finland where according to unofficial commercial, statistics, the annual consumption of PCB is about 2^0 metric tons (Helminen, 1970). PCB containing 20-50# chlorine increase the oxidation stability and eliminate flammability of mixtures of mineral and silicone oils (Marek et al.j I960). A trichlorobiphenyl preparation (Pyranol l*+99) may be used to prepare media with high dielectric strength (Cox, 1970). PCB are one of the Ingredients for coloring of molten linear polyesters (Brinkman, Jr., 1969)- Details of the industrial preparation of PCB in the U.S.S.R. were published (Trofimov et al., 1969)* The crude chlorinated product contains 1-2.5# hydrochloric acid and traces of ferric chloride. The latter forms complex compoundst diminishing the Insulating properties of the preparation. The impurities can be removed by treatment of crude PCB with sodium hydroxide at 150-170 C and distillation. This is very likely the process generating highly toxic chlorinated dibenzofurans found in some PCB preparations (Vos et al., 1970b). The preparation of 3 , 3 `,5>?," tetrachloro-2.2l ,6,6*-hexafluoro- biphenyl was described (Ishikawa, 1969) and polymothylated pentafluorobiphenyls were prepared (Brown et al., 19&9). The use of mixed chloro-fluoroblphenyls in hydraulic fluids with Improved heat stability, fire resistance, lubricity, viscosity, and anti-corrosive properties was patented (Boschan et al., 1970). Another patent describes tho preparation of diamino- polyhalobiphenyls (halogen: chlorine, bromine) to be used as starting materials for the preparation of flame-resistant polymers and hardening agents for epoxy resins (Sobel et al., 1970). A new series of unsaturated, flame-resistant polyester resins, based on diols derived from decachloroblphenyl was developed (Anon, 1971). The preparation of bis(chloromethyl)-octachloro- blphonyls as intermediates for manufacturing dyes, plastics, NEV 036109 -i* ~iniii i h.iWlifti ^ &-- 3 * ,* or plant protective agents was patented (Woppart et al., 1970). The flame retardant additive, Firemasttr BP& Is hexattromobiphenyl (Michigan Chemical Corp,, 1971)* PCB may be used for quenching of heated metals (Massey, 1971) Aroclor 1221 was used in preparations increasing opacity and brilliance of fibrous and paper substrates (Vacslliades et al*, 1 9 7 D . Chlorinated biphenyls and trichlorobenzene are the insulating liquid in General Electric Pyranol transformers, induction voltago regulators and exciting-current interrupter switches* Tho first transformer Pyranol was introduced in 1932. It was later modified by the addition of tetrnphenyl tin, used as hydrochloric acid scavenger* The present-day formulations contain diepoxido which has several advantages over tetraphenyl tin (General Electric. 1967). Trichlorobenzene is sometimes added to PCB preparations to lower the solidification temperature (Gambaro et al., 1969). A typical preparation contains **0# of trichloro benzene and 60# of hexachloroblphenyls (Jay. 1970). The impregna tion of polypropylene with 5-20# trichlorobiphenyl and 0,1-10# A epoxide stabilizer to produce dielectric films on capacitors was patented (Eustance, 1 9 7 D * Traffic markings may contain PCB (Japan Paint Co., 1970). The preparation of isomer-free p,p'disubstituted biphenyls from substituted phenylmercurie acetates was patented (Kraus et al*, 1970). Polymeric products of unknown composition, possibly incorporating oxygen, are formed from PCB on heating. The addition of organic phosphites such as diisopropylphosphite, and nitrogen or carbon dioxide atmosphere were used to suppress this reaction (Sullivan, 1971). The synthesis of some nitrobromobiphenyls was described (Dell'Erba et al., 1971X and the preparation of 3>5i 3* J'-tetrabromo-P,1*,2* ^-tetrahydroxybiphonyl, Tebrophen was patented (Pershln et al., 1971. This compound 5r. an antiviral agent, effective particularly in viral eye infections (Nikolaeva et al., 1970). Routes for the synthesis of individual chiorobiphenyls with k and less and with more than * chlorine atoms, with special attention to the preparation of rndJoactlvely labelled chloroblohenyls were discussed (Melvas, 1970; Wachtmeister et al., 1970). The preparation of ^Cl-labelled PCB of the Aroclor 12h8 and 125^ type for tracer studies was recently described (Stalling et al.. 1971). More than 20 chlorobiphenyls were recently prepared (Hutzinger et al., 3971). Disposal techniques for PCB-containing wastes were discussed. PCB can be destroyed by incineration at temperatures above 800C (Peterson, 1970). Chlorinated 3-pentadecenyl phenyl methyl ether can be ' usod as plasticizer and extender for polyvinylchloride (Ghatge ot. al., 1970). Chlorinated naphthalenes are another.group of Industrially important chlorinated hydrocarbons. In the U.S.A., they are manufactured under the "tradename Halowax (Anon.), ilalowax 1000 is a mixture of mono- and dichloronaphthalones NEV 036110 j.', <*.-..u.. t a t j u i .'-.viAt->A .m. s . -** -*>*-*rtW *iK 'A - - r. . 1i l u. ! containing 26$ chlorine and is used mainly in motor tune-up j compounds and upper cylinder lubricants Halowax 1031, mostly I 1-chloronaphthalene is of better purity and.has similar uses. Halowax 1001 and 1099 are mixtures of tri- and tetrachloronaphthalenes with a chlorine content of 50-52$, and are used as dielectrics, binders for electrical grade ceramics and paper coatings. Halowax 1013 and 101U contain from U to 6 chlorine atoms per molecule (56-62$ chlorine) and are used as insulators, oil additives, and flame retardants. Halowax 1051, basically octachloronaphthalene (70$ chlorine) may be used as a component of organic fillers where flame retardancy is required. A chlorinated hydrocarbon preparation, Cereclor 552 . (mol. wt. ^00, 52$ chlorine) was used to flameproof polyacrylo- . nitrile fibers (Torrance et al., 1971)* Chlorinated paraffins with 15 carbon, atoms in the molecule, containing 50$ chlorine were used to prepare dispersionstable polyvinylchloride latexes (lida et al., 1971). .Chlorinated paraffins are also used in paper-sizing agents (Chiba et al., 1970), and probably the main application- o f 'these compounds is in the flameproofing of materials. Chlorinated paraffins used for . this purpose contain *+0- 50$ chlorine and 20-2M- carbon atoms in the molecule (Anon., 1969). Chlorinated paraffins were used in flame retardant thermoplastic ethylene-vinyl acetate copolymer i compositions (Kosaka et al., 1970). Chlorinated-paraffins I together with zinc fluoroborate and a nonionic emulsifier were i used to Improve the fire resistance of acrylic textiles (Fidell, 197D. Flame retardant compounds may also be chemically incorporated into polymers. Such-compounds contain, in addition to halogen atoms, one or two functional groups which can participate in the polycondensation reactions. Presently used compounds Include chlorendic anhydride [1 ,*1-,5*6,7,7"hexachloro~ b!cyclo-(2 ,2 ,l)-5-heptene-2 ?3-dicarboxylic anhydride], tetra- chlorophthalic anhydride, "etrabromophthalic anhydride (Firemaster PHT1!), and dimethyl-di-(3,5~dibromo-4-- hydroxyphenyl)- methane (Firemaster BPkA, Michigan Chemical Corp.). Other compounds suggested for this purpose are 3>3*,5,i '^-tetrafluoro" biphenyl-^ W1-dicar boxylic acid, 2 , 2 '^j^-tetrafluorobiphenyl*' 3>3'~dicarboxylic acid, 2,2',lf,l+'-tetrafluoroblphenyl-5,51*' dicarboxylic acid, 2 , 2 1,6,6'-tetrafluoroblphenyl-33'-dicarboxylic acid, 2,2* 1,6,6 '-hexafluoroblphenyl^^* -dicarboxylic acid , and 2,2*,5,5,-tetrafluorobiphenyl-6,6i-dicarbovylic acid (Sugawara et al, 1971). Fumarate and maleate polyesters of 2,2',3,3`y 5,5 ,6,6,-octachloro-l<.k,-(2-hydroxyethylene)-diaminobiphenyi and 2,2" ,3,31.55* >6,6*-octachloro-H,k '-bis(diethylenedioxy; biphenyl have excellent flame resistance (Vogt et al., 1971)* Chlorinated fatty acid esters are used as low-cost plasticizers for a variety of resins and plastic materialsMethyl dichlorostearate and Plasticizer MSP-500 (Anon., 1969) are NEV 036111 examples of this class of compounds. As a rule, brominated compounds are about twice as offoctlvo fire retardants as chlorinated compounds. Aliphatic bromine compounds are about 1 .5 times more effective than aromatic bromine compounds (Schneider et a l . f 1969). Brominated vegetable oils are used as haze stabilizers in citrus-basod soft drinks. Chlorinated polymers such as chlorinated polylsoprene, , polyethylene, and polypropylene, with a chlorine content of m O-60# are used as binders for paints and combination resins for printing inks (Hoehne et al., 1970)\ Halognation of polyesters for the preparation of self-extinguishing films was patented. Such preparations contained 6-25# bromine and 3-6# chlorine (Jackson et al., 1971) Perfluorinted hydrocarbons have low boiling points for their molecular weight, extremely low surface tension, and very high density and viscosity. They are poor solvents for most compounds except gases. Perfluorinated hydrocarbons have an excellent chemical 'and thermal stability * and very low toxicity (Sargent et al., 1970). Methylene chloride (10-80#) and perchloroethylene (0-20#) are used in heavy duty dJphase cleaners and a method for their determination by gas chromatography was published (Bennett et al., 1971) Tri- and totrachloroethylene mixed with surfactants to form water-in-oil type of emulsions can be used for cleaning ceramic, metals, and other materials (Johnson, 19 7 1)-- Other halogenated hydrocarbons are used as solvents, ftamlgtmts, refrigerants, aerosol propellants, etc. A list, of the more-Important industrial halogenated hydrocarbons is Included In Appendix. Contamination of the environment by industrial halogenated hydrocarbons FOB are used In a wide variety of products and hence may enter the environment via different routes. Direct spills or leakages from heat exchange systems are one such route and occurred, for example in Escambia Bay, Florida (Duke et al., 1970)v General aerial fallout may be another route since particularly the lower chlorinated biphenyls are appreciably volatile. Sewage outfalls and ocean-dumped sludge are probably major sources of PCB. Concentration of PCB in raw sewage from the Glasgow area ranged from 0.1 to l1! ppm as Aroclor 12$*i on wot weight basis, and it was estimated tnat the PCB discharge into the Clyde estuary is of the order of 1 ton per year. Similar estimates were obtained for the Manchester area and the Tlinmos estuary (Holden, 1970a). In California, sewage outfalls discharge from 0.06 to 0.7 tons of PCB per year in water and h ' a t 1 i d t * --,'2 .& & f toJ ^ n A m a u ^ ^ i j i i ,- ^ t i.- f u i , 6. from 0,6 to 36 tons per year In sludge (Schmidt et al., 1971). Low molecular weight chlorinated paraffins and olefins, such as 1 ,2-dichloroethane, 1 ,1 ,2-trichloroethane, symmetrical and unsymmetrical tetrachloroethane, pontgtchloroethane, chlorinated butanes, cis~ and trans-isomers of chlorinated 1 -, and 2-butenes are byproducts of vinylchloride production by the oxychlorlnatlon process. Large quantities of these compounds are dumped Into the sea and the hydrocarbons were recently detected In fish and water samples'from the Norwegian, Barents, and North Sea (Jensen et al., 1970). Only limited data on the occurrence of other halogenated hydrocarbons In the environment are available. A variety of halogenated hydrocarbons is released into the atmosphere, from where it may reach the aquatic environment. For example, about 3*6 x 10 ^t/year of tetrachloroethylene evaporates from dry cleaning solvents in the U.S.A. (Goldberg, 1970). Fluorodichioromethane, difluorodichloroethene, fluorotrichloromethane, difluorochi oromethane, difluorodichloromethanetrifluorotrichloro'ethane, mothylchloroform, vinylidene chloride, trichloroethylene, tetrachloroethyleno, chlorobenzene, and dichlorobenzene were detected In the atmosphere of the U.S.S. Hammerhead (Saafeld, 1971), which may serve as a model of the earth1s-atmosphere. Some decomposition products of the above-mentioned hydrocarbons may also exist in the atmosphere. Thus the very toxic dichloroacetylene is generated by alkali from 1 , 1 ,2-trichloroethane (Saafeld, 1971)* Hexachlorobutadieno (132 mg/1), tetrachloroethylene (15^ mg/1 ), trichloroethylene (65 mg/1 ), and carbon tetrachloride (29 mg/1 )`were detected by gas chromatography (5# tricresyl- phosphate on a diatomite at 130C) in waste water from the production of hoxachlorobutadlene (Mikhallok et al., 1970). An Industrial mixture containing hexachlorocyclopentadiene was analysed by gas chromatography on silicone E-301 liquid phase at l6oCc Octachloropentadiene, hexachlorobenzene and unidentified Impurities were also detected (Dzhioeva et al., 1970). A maximum allowable concentration of 0.31 mg/1 was recommended for allyl chloride in reservoir water (Karmazin, 1966) . " The suggested maximum allowable concentration of tetra- chlorobenzeno is 0.02 mg/1 (Fomenko, 1965), and that of trlchlorofluoropropane is 0.1 mg/1 (Selyuzhitskil, 1963). Suggested maximum permissible concentration of carbon tetrachloride in water reservoirs is 0.3 mg/1 (Kuteyov, 1968). From 18 to 1*2 ^g/1 of organically bound chlorine was detected In Rhine water, h 5 Mg/1 In waste water and 25 Mg/1 In biologically purified waste water. Most of the organochlorine compounds wero adsorbed on suspended matter. The average 1969 content of these compounds in suspended matter, collected from the River Rhine at Dsseldorf was 66 mg/kg on ary weight basis (Koppe et al*, 1970b). N6V -36li3 Analytical methods for PCB. chlorinated n a p h t h a l e n e a n d polychlorinated terphenylBi. and other halocenatcd hydrocarbons PCB accompany, chlorinated hydrocarbon pesticides in the usual extraction and cleanup techniques and both groups must be separated before quantitation by gas chromatography. Column and thin-layer chromatography are usually used for the separation, PCB were eluted from Florisil columns with hexanes DDT and other pesticides with mixtures of ether and hexane (Reynolds, 1969).. Another laboratory had difficulties with this technique and reported that DDT was already partly eluted with hexane (Bevenue et al,, 1970), Column chromatography on silicic acid separates PCB from all common chlorinated hydrocarbon pesticides except p,p*-DDE (Holden et al*, 1969), and under very carefully controlled conditions even from p,p'*-DDE (Armour et al,, 1970), Some carry-over of PCB Into the pesticide'fraction was demonstrated by a combined GLC-MS procedure when extracts containing more than 0,*4 g fat were placed on the silicic acid column (Bellman et al,, 1971) Small amounts^ of aromatic hydro carbons such as benzene, present in some pesticide-grade hexanes affect significantly the elution patterns of PCB and organochlorine pesticides from silicic acid columns, and may be responsible for the discrepancies reported above (Zitko, 1971a)* Thin-layer chromatography on alumina with n-heptane as the developing solvent was used to separate PCB from organochlorine pesticides. Areas'on the plates, containing the respective compounds were extracted with methanol-hexane and the extracts were analysed by gas chromatography (WestOO et al,, 1970c),, TLC on alumina, coated with silver nitrate, was used to somi-quantitatively determine PCB after oxidation of p,p'-DpE to *4,>4'-dichlorobenzophenone with chromic oxide in acetic acid. Developing solvent was 5% benzene in hexane and the commercial PCB preparations Arocl or 12*42, 12^8, 125^, 1260, and 1262 moved os r/inglo spots with R- 0.91'-0.9l* (Mulherh et al,*, 1971 ) A TLC technique based ona two-dimensional development with n-heptane and n-heptane-acetone 98;?, respectively, was described (Fehrlngcr ct a l , 1971)* PCB components can be effectively separated by reversed-phase TLC using IIquid-pa raffin impregnated silicic acid as the adsorbent and a mixture of acetonitrile, acetone, mothanol, and water, ^OslB^+OsZ, as the solvent (De Vos et a l ,, 1971)o A method based on reversed liquid-liquid partition between water and Chromosorb W, coated with n-undecane and Carbowax *4000 was developed for the determination of PCB in water,, The sensitivity of the method is in the ppt range (Ahling et al,, 1970). Gas chromatography of chlorinated biphenyls on capillary columns coated with polypropylene glycol 750 was described (Woingarten et al,, 1962;, and an excellent separation of mono-, dis and trichlorobiphenyls by gas chromatography on polyphenyl NEV 03611* ^*^1.waw luiuuti.. **/.Sfcwiu.Ls. 8. thioothors was patented (Emery et al., 1970)* Various other phases aro used for gas chromatography of PCB. One of the most frequently encountered is the silioone grease SE-30, often mixed with other materials such as QF-1. Electron-capture detector response to mono- and dlchloroblphenyl was reported (Gregory, 1968). The response to mono-, dl-, tri-, tetra-, penta-, hexa-, and octachlorobiphenyls, and to decachlorobiphenyl was recently determined (Zitko et al., 3973a). The detector response strongly increases with increasing number of chlorine atoms in the molecule* For example, the responso of decachlorobiphenyl is 500 times stronger than the response of ^-chlorobiphenyl. Host of the increase occurs in the mono- to trichlorobiphenyl range and the response Increases only by a factor of 2-3 between tetra- and decachlorobiphenyl. The detector response depends also on the chlorine substitution patterns. Thus chlorine substitution in positions 2 and 6 decreases, and vicinal substitution, particularly in positions 3 and *4 Increases the electron-capture detector response. PCB found in wildlife are of the Aroclor 125^ or 1260 type; according to mass spectra, their components are tetra-, penta-, hexa-, and hoptachloroblphenyls (Kooman et al., 1970), and the detector response to these compounds is not likely to vary more than by a factor of 2, which is a reasonably good basis for quantification, even without detailed identification of the individual chlorinated biphenyls. Carbon skeleton chromatography of Aroclor 1260 was recently described (Asai et al., 197l). The preparation Is converted essentially into a mixture of cyclohexylbenzene and biphenyl at the catalyst temperature of 300, and into cyclohexyl- benzeno and bicyclohexyl at 260. The technique may be useful for confirmation purposes. Computer-controlled gas chromatography-mass spectrometry was used to identify tetrachlorobiphenyls in a Los Angeles sewer extract and in a sturgeon ovary extract (Bonelli, 1971). Randomization of chlorine atoms between phenyl groups occurs in the electron impact-induced fragmentation of PCB and with only one exception (2,2'-dlchloroblphenyl) the primary ion spectra of different isomers are virtually indistinguishable which means that the use of mass spectrometry for structural studies of PCB is limited (Safe et al., 1 9 7 D . Reactions of decachlorobiphenyl were recently described. Decachlorobiphenyl reacts as substituted pentachlorobenzene in nucleophilic substitution reactions occurring preferentially at the h ,m '-positions (Binns et al., 1971) Some of the reactions could possibly be developed into confirmatory tests in residue analysis. Thus decachlorobiphenyl yields a mixture of h and ^ ^ '-substituted nona- and octachlorobiphenyls on reduction with lithium aluminum hydride or butyl lithium. The reaction of decachlorobiphenyl with sodium methoxide in boiling pyridine yields a mixture of mono- and dimethoxy nona-, and octachlorobi p h e n y l. NEV 03611* rfl -*** LS4* ' --k *'>i * < iJj fcifc.i A * 1 1 9. Many peaks In gas chromatograms of PCB and the lack of Identification of Individual chlorobiphenyls make the quantification of PCB difficult, since both height and area ratios of PCB peaks In environmental samples are usually different from those In commercial PCB preparations. Some authors use total peak area (Armour et al., 1970) or.area under one (Koeman ot al., 1969), two (Vermeer et al., 1970), or more peaks (Kell , et al, 1971; Prestt et al., 1970), others use peak heights (Grant et al., 1971; Hansen et al*, 1971; Rlsebrough, 1969)* The valuos thus obtained are compared with values obtained in the same way on commercial PCB preparations (usually Aroclor 12JU). A method based on the average electron-capture detector response to biphenyls containing from 1 to 7 chlorine atoms using commercial Aroclor preparations as standards, was recently described ( Rote et al., 1 9 7 J K The method is not more accurate than any of the other quantification procedures, since the detector response to chlorinated biphenyls with the same degree of chlorine substitution may vary significantly according to the substitution patterns (Zitko et al*, 1971a)* Although some believe that the quantification of PCB with any degree of confidence is impossible (Richardson et al., 1971), the general opinion is that the quantification is reasonably precise. The results obtained recently by different laboratories on an exchange sample of PCB, included In the O.E.C.D. pesticide monitoring program, have the name order of variability as usually occurs in the determination of chlorinated hydrocarbon pesticides (Holden, A. V., personal communication). However,, because of .the lack of Individual chiorobiphenyl standards, the quantification cannot be accurate. Wo definite quantification procedure for PCB is developed and "laboratories are encouraged to exercise good judgment in their measurement of PCB residues and to explicitly describe the analytical procedures employed when reporting residues of PCB" (Burke, 1971). ^ During the analysis for PCB, great care must be taken to avoid contamination of samples, since PCB may be present in n variety of products. Thus PCB of the k0% chlorine variety were found in cardboard (Bailey et al., 1970) and in a wrapping .plastic material (Mestres et al., 1970). No analytical methods are available for the determination of halogcnated biphenyls containing different halogens. PCB are very likely metabolized by hydroxylation. Microbiological metabolism of biphenyl (Lunt et al., 1970) and the metabolism of k^chlorobiphenyl in the rabbit (Block et al., 1999) have been described, and methods for the analysis of the phenolic metabolites of biphenyl (Raig et al., 1970) and of dichiorobiphenyl diol isomers (Toraorl, 1970) by gas chromatography wore developed. In the latter case, silylation was used to block the hydroxyl groups. On polyethylene glycol adipate the elution sequence was S^'-dichl o r o - , SjS'-dichloro-, 5,5'-dichlpro ?,;?' hydroxybiphenyl NEV 036116 *4L - * i * ; ifca *J p 10. The constituents of Aroclor 12*f2, 12JU, and 1260 have been recently Identified by gas chromatography, NMR spectroscopy, and mass spectrometry. Aroclor 12ffa yielded 69 constituents and the major components were identified to be 2.5- 2' , 5 1-, 2,3-2',5'* and 2,5-31 tetrachlorobiphenyls, 2 .5-2 ' , 3 ' , .5-?',3'^'-, 23,,3i--22*.,3,13,,6,6''-,-,2.55.-U2-'2,TH ' , 5'-, 2 ,kr2 * ,5 ,5'", ana 3,^-i' 1*') ,3? pontachlorobiphenyl, 2 ,3 ,6-2 *,h ',5'" , 2 ,3 ,^-2 *,3 *,o'-, 2,li,5-2 ',5 1-, *nid 2,3,^-2' ,**' ,5'tanexachlorobiphenyi. The fractionation of Aroclor 12^2 ana 1^60 yielded ^5 and 78 components, respectively. The most frequent substitutions in Aroclor 1 2 h2 occurred in positions 2-, 2,*+-, and 2,5- The structural features of Aroclor 1260 components were similar to those of Aroclor 1251* but the preparation contained more heptachloro biphenyls (Sissons et al., 1 9 7 D - Kovats retention index system (HI) was used to characterize PCB in this study. As already mentioned Jn terms of retention times, substitution in ortho positions decreases HI. Chlorinated naphthalenes resemble PCB in Industrial uses and chemical properties. Chlorinated naphthalenes accompany PCB on silicic acid chromatography (Armour et al. , 1 9 7 D - Chlorinated naphthalenes are eluted in the same fractions as PCB ( and II) also on the alumina-silica chromatography of Bolden and Marsden. If present alone or in a concentration comparable to that of PCB, their presence could possibly be recognized from the characteristic peak patterns on gas chromato graphy (see Appendix). UV spectrum of a commercial mixture of chlorinated naphthalenes has a naximum at 306 nm with an 1% A lcm 329 (s^e A PPndi.x). PCB (Zitko, 1970) and polychlorinated terphonyls (POT) have only negligible absorbance at this wave length so that chlorinated naphthalenes could be confirmed in the presence of PCB and PCT by UV spectrophotometry. The limitation for this method is the background of UV-absorblng impurities eluted from alumina and silica columns and UV-absorblng compounds present in biological samples and eluted in fractions I and II. PCT are not eluted from 5# DEGS-2# orthophosphoric acid (Hcynolds, 1971) and SE-30 columns at 200C. PCT (Aroclor 5^60) can be determined by gas chromatography on a 6 ft x W mm glass column containing 3# 0V-210 (Pierce Chemical Company) on Chromosorb WAW 60/80 and operated at 200C. Under these conditions Aroclor 5^60 gives 8 major peaks with retention times of 1 .78, ?.1<S 3.1 0 , 3.96, ^.83, 5-88, 7-25, and 9.13 relative to docachJoroblphenyl, respectively. The electron-capture detector response to Aroclor 5^60, based on the sum of heights of peaks 3.10, 3.96, and W .83 is 0.122 per ng relative to docachl oroblphenyl. More than 90# of Aroclor 5*+60 is eluted in fraction II. Using this technique PCT were detected in some c*nvl ronmental samples and attempts are underway to confirm this finding by mass spectrometry (Zitko et a l . , 1971b). NEV 036117 o i ' A m v (M4 . f a Ka tA t f c - lifc if b t t . o l t ' . v j f t i a j t ' k A * li.j* < i> f i j f r t w r * s * - . .w* A i> i u- . . ^ . - 11. Flrcmaster BP6 (hexabromobiphenyl) gives 7 peaks with retention times of 0.50, 0.88, 1.0 9 (major peak), l.^H, 1.77 1 .9^1 and 2.82 (major peak) relative to decachlorobiphenyl on tho 0V-210 column. In the pesticide cleanup chromatography the hexabromobiphenyl preparation is eluted In fractions I and II, the two major peaks being eluted mainly in fraction I (Zitko, V . , 0. Hutzinger, and P.M.K. Choi, unpublished). The determination of chlorobenzene In water was described. The compound was extracted with methylene chloride and determined by gas chromatography (Martur et al., 1968). * Chlorobenzene was removed from waste water by gravity separation, extraction with dichloromethane and adsorption on activated carbon. Residual dichloromethane was removed by aeration (Radvinskii et al., 1969) Methods for the determination of carbon tetrachloride In water were reviewed and a method based on the extraction of 4 carbon tetrachloride with methyl ethyl ketone and its color i ' - reaction with pyridine In an alkaline medium was suggested (Stankov!c, 19&5). : The presence of nonvolatile organochlorine compounds' < which cannot be determined by gas chromatography in hexane extracts of environmental.samples could be detected by the decomposition of samples with metallic sodium and determination < of chloride as described for some chlorinated hydrocarbon ; pesticides (Koblitsky et al., 1962). ti A semi-quantitative method for the detection of less-volatile lipophilic chlorine compounds by circular TI and development with silver nitrate was described (Koppe et al., 19?0a). PCB levels in the aquatic environment The presence of PCB in the environment has been unequivocally established. Of major Interest at the moment are comparisons of PCB levels in different geographical areas and the sources and transport mechanisms of PCB in the environment. Levels of PCB in fish, seals, and birds are summarized" In Tables I and II. PCB levels in eggs of aquatic birds from the Bay of Fundy and Passamaquoddy Bay are presented in Table 111. Some of the data have already been mentioned in the PCB reviews, but are presented again for comparison purposes. The analyses of fish Indicate ubiquitous occurrence of PCB, with higher levels in fish from coastal areas and enclosed bodies of water such as Gulf of Maine-Bay of Fundy, Archipelago of Stockholm, and the Baltic Sea. Relatively fewer data are available for freshwater fish but Is very likely that the trend is similar, and fish from localities close to industrialized area! and population centers contain elevated levels of PCB. Tho accumulation of PCB in the food chain Is indicated by the NV 036118 9 II i 12. levels of PCB found in seals and in aquatic birds. The levels of PCB in eggs of cormorants, herring gulls, and black ducks from the Bay of Fundy and from Passamaquoddy Bay are surprisingly high and so are the levels in the body tissues of one cormorant and one herring gull from this area. The other herring gull, found I dead on Hospital Island has lower residues of PCB and p,p'~DDE Jn the tissues. This indicates that there may be a-considerable variation of PCB levels between Individual specimens. The concentration of PCB in one cormorant from The Netherlands is very high. It is interesting to note that PCB and p,p'-DDE are present practically exclusively in the yolk. For example, D.C. cormorant eggs.from Station F contain 92.? g/g of PCB and 6l,8| Mg/g of p,p'-DDE in the yolk, while only 1.37 /xg/g of PCB and 0 tg/g of p,p'-DDE were found in egg-white. The toxicological significance of PCB levels in the environment cannot be evaluated at the moment. The levels of organc-chlorlne pesticides and PCB in fish, margarine, vegetable oils, and other products from Sweden have been reported (VJestBD et al., 19?0a,b). Data on some terrestrial and aquatic organisms from Norway wore published (Bjerk et al., 1970). Three samples of commercial dry fish food, analysed lln this laboratory contained 0,15, 0.13, and 0,25 /g/g of PCB, respectively. This may represent an important source of PCB in laboratory- and hatchery-reared fishes. For example, one-year-old Atlantic salmon-parr, reared at this Station contained 0,20 fig/g of PCB in liver and 0.88 tg/g in caeca, while small parr contained 0.30 /g/g (whole fish). Hatchery-reared rainbow trout (Salmo galrdnerl). held at the Station for experimental purposes, contained 0.14 0.22, and 1.49 /ig/g of PCB in muscle, liver, arlu caeca* respectively. (All concentrations are given on wet wei(ht basis). The tissues of both salmon and trout contained only traces of p,p'-DDE. The levels of PCB may bo significant if tfje fish are used in testing sublethal effects of PCB and other pollutants. Three samples of Canadian margarine were analysed in this laboratory and PCB concentrations found were 0,47, 0.21, and 0.18 ^g/g for margarine containing 80# marine oils, 60# . marine oils, and 100# vegetable oils. The levels of p,p'-DPE were 0.07, 0.2 5 , and less than 0.01 jxg/g, respectively. ed NEW 036119 \ 6310 v J \\ H6&* , ] \ 398&M Table I, PCB and chlorinated hydrocarbon pesticides in fishes and fish-eating birds. 13. Species Bluefln tuna (muscle) Thunnus thynnus (liver) (muscle) Location California New Jersey PCB, p ,p 1-DDE, jug/g jzg/g 70* 0.2?* O.O1** *0.09* O .36 0.0m- Other Yellowfin tuna (liver) Ihuanus fllfra&ELtfes (") (") Skipjack tuna (liver) Buthynnus nelamls (muscle) (liver) (liver) (muscle) (muscle) Bluefish (muscle) Pomatonus saltstrlx Galapagos 40.01* Central America O.Ok-.. West Africa 0.17 Hawaii * 0.10* 0.01* 0.19** 0.03 0.01* Hawaii Galapagos Ecuador New Jersey West Africa < 0 401* 0.01* <0.01* <0.01* <0.01* 0.82** 0.12** o . m * * 0.02** New Jersey o.eif** 0.17** DDT 0.07** Swordfish (muscle) yiphlas, jsladJlus Blue marlin (muscle) Makalra nigricans Cay Sal Bank Walker's Cay 0.22** 0.02** 0.16** 0.06** Northern anchovy (whole fish) California Bn'graulls mordax 1.00* 11.6* Horring (whole fish) Eastern Canada Cluoea haremrus (muscle) Baltic Sea Archipelago of Stockholm Gulf of Bothnia O.UO*** 0 13*** 0.27 o '.28+ . i 0.12* o.o6* 0.12+ DDT 0.09*** DDT 0.1+0+ DDT 0.11* DDT O .lk * Sprat (whole, dry wt. basis) filupaa sprattus Dutch Wadden Sea 0.3-2.01* Pacific jack mackerel (whole fish) XfaChUCUS lM!ill California 0.02* 0.32* Mackerel * (muscle) Bay of Fundy Scomber scombrus 0.35*** 0.07*** NEV 036120 * ++ *0? *+* :* :i* 'ichM&<* ********^ -ww. -vj-j-^t-j w.tv.. jt 1U, (Table I Cont'd) .Species Location PCB p,p'-DDE m e /e Hg/ Other Shiner perch (whole California fish) Cymatogaster aggregate English sole (whole fish) Parouhrvs y.fltulttB California Plaice (muscle) Mova Sootla Hlupoelossoldes platessoldea Bay of Fundy 0.90' 0.37 6. 08*. O.18* *** 0.03 0.01 *+ + 0.38** 0.01 *# Plaice (muscle) ElSM SM &S&ZSi plaifi^aa (muscle) Swedish West Coast Baltic Sea Hake (whole fish) California ilcUuco.lus .pf-gfluot.ua 0.02 0.02 0 . 12' <0,01 <o.or 0.12* White hake (muscle) Nova Scotia Banks Bay of Fundy 0.02* O.W*** 00..0032***** Ocean perch (muscle) ftebas tes rqarinus Nova Scotia Banks<0.01***<0.01*** Bay of Fundy 0.32 0.03 Cod ' Oadus morhua (muscle) (muscle) (muscle) Nova Scotia Bonks 0.02*** 0.01*'** Bay of Fundy 0.55** O.oV** Swedish West Coast Baltic Sea 0 .02* 0.03+ o.or 0.03* Sea ravon (muscle) Bay of Fundy Hemltrlpterus amerioanus (viscera) Bay of Fundy 0.21 +* 0.08 ++ 0.73 0.30** DDT 0.2U Picked dogfish (muscle) Swedisli West Squalus acanthlas Coast 0.1?* 0.06* Atlantic salmon Salmo salar (muscle) Eastern Canada ** O.hJ (muscle) Tagged in Canada. ** (caught in Greenland) 0.20 Sraoit (whole fish) Eastern Canada +* 0.10 0.22** 0.0V** 0.28** NEV 036121 f - 9* [* j n 1 i MJ***--<>-_kW>U.*- -_'.... 15. (Table I Cont'd) Species Location PCB p ,p 1-DDE >ie/g ne/g pthor Adult salmon (muscle) (liver) (caeca) (ovaries) (testes) (muscle) Eastern Canada Baltic Sea 0.62** 0.8** 0.65** 0.7 1" 0.371* 0.30+ 0.07** 0.1^** 0.12** 0.1 1 ** 0.<*** 1.9+ DDT 1.5* Commercial fish oil Eastern Canada Baltic Sea Swedish West Coast 3.55** 3.5* .^ 0.7V 2.27** 8.7* J, 0.9 DDT 0.3?** DDT 7.3+ DDT 1.2+ Cod liver oil 0E*CD test 1.39* 0.51* DDT l . U V Mussel (whole) Mxtllus eflulls Swedish West Coast 0.08 O.Olt Baltic Sea 0.03+ 0.01+ Archipelago of Stockholm Eastern Canada Finland o.ov.. *02!** O.lV*** 0.02*** 0,06* traces Holland O.V Norway traces Sweden 0.03* Dogfish pqualns flcanthlas Coho slmon .OncorhynchuS: klsutfill Flounder (a.fallshthy& sp) Norway Sweden Lake Michigan Escambia Bay o.oi-o.ov 0.15* l`t.6++ 10.V** DDT 3.2+ w5*++ Croaker Mc.roppftpn nfluJLfttus Escambia Bay 12 +++ Menhaden^ . l&axQgtfrlft patronas Escambia Bay 11 +++ Plnfish Lagodon rhombodes Escambia Bay +++ 10 Speckled trout CxaP.SpJ.9Jl nebulous Escambia Bay 20>+++ Shrimp Ponaeus setiferus and Escambia Bay 1.5-2.5+" NEV 036122 > J.i r f 2* .Mr .2** J r t.' w A m i r i > i f t V .te A . ^ A v ^ * . t - -. ^ .. . L , `. . . . ,, M L * M t * * * . ,, > * .. ..-, 16. (Table I Cont'd) Species Location PCB P jP ^ D D E fxg/g Mg/g Other Blue crab Calllnectes saoldus Escambia Bay American eel (muscle) Eastern Canada tiiR vLU a xsaif-ata (liver) 1.7 0.71*** 0.50*** 0.57*** 0.19*** Chain pickerel (muscie) Eastern Canada fisox nlr.er (liver) DDT 0.63 Pike Esox luciua Holland Norway Sweden 0,05* traces* 0.02* Seal (blubber) Pfcosa vltullna and iM i p h Q 9 .rug. Baltic Sea ' 15* Archipelago of , Stockholm 6.1 3>t+ 3 1 .8+ DOT 32+ PDT 1+.2 Grey seal pups (blubber) S.W. England 1600* (on U p l d basis) N.E. England 501 Sable Island 12" Orkney Islands 79 Grey seal adults Sable Island 20* Orkney Islands 17" 68fA.f= 5.*e 3.1" 22 _ `t. T PDT 3f , 7 .1 : 5.**: 3.1 171 r *Hisebrough, 1969 *+This report ** ZltKo, 1971b * +Jensen et al., 1969 Holden, 1970c **Armour et al., 1970 ++4Duke et al., 1970 =Holden, 1970b ^ i i 17 Table II, PB and p,p*-DDE in birds. , \ - * Species location Mg/ PCB p ,p'-DDE Brandt's cormorant Phal&crocojax pcnlcJllatns (eggs) Farallon Island 113 + * 326+ * Pelagic cormorant (eggs) California . polap.lcus 62+* 128+* Bulked shag . . (eggs) Britain, West Coast . arlstPtolls .. Brltlan, East Coast V* D.C. cormorant . aurltns (eggs) i North Dakota Wisconsin Manitoba Western Canada 3-** 25! 10= 0.672.21* !:!" JF 20= ' 3.3-3.5- Laru^rfl3 Razor bill AicA torda (eggs) (eggs) Alberta Western Canada Britain,West Coast " East Coast 0,87+= 7.26+" X.57-1.7W 2.9-21.8 o++ 6++ K I W I wake . >.> (eggs) Rlssa trldactyl a %* I* Mallard duck nos ulatyrhynchos (eggs) West " East " Western Canada 8++ 3++ 0.09* 0.2* Guillemot Dreg aalfiftfi (eggs) BritainWest Coast 11 East " Baltic Sea 8++ 3++ J6* 2.0** 0.7 38.8** While pelican .Polec.anus' orvlhrorhYnchos Common tern Sterna hirundo (eggs) (eggs) North Dakota ) ) Saskatchewan ) California South Carolina Florida Western Canada 2s 1= 8-12*** 8-12* 3-8* 76-135* ** XX,.. 2-7* 0.9^-1.57* 5.1-33.3 Great Blue Heron rda herodlas (eggs) Western Canada trace-31*!* 3.7-78.0 NV 036124 Vs s 18. (Tabi II cont'd) Species Location Mg/g PCB p,p*-DDE Whitotalled eagle (Pect. Hallaectus plblcllla muscle) Archipelago of Stockholm 190** 330** Heron (Pect. muscle) ftfdoa cinerea Archipelago of Stockholm 9,^00* * 1^,000* * Common cormorant (total body) P. .carbo The Netherlands D.C. cormorant (muscle) Passamaquoddy Bay aurltus (liver) Subcutaneous fat Abdominal fat 0 Herring gull (muscle) Passamaquoddy Bay . arr.entatus (liver) Subcutaneous fat Ovaries k02** llf.3** +** 32-.1338t*** 8.1*0*** if.16*** 38*<*** 16k*** 52*** 162*** . 5.O6***' 2.07*;" (2.69 )C0.75 *) 6.50*** 2.08*** (1.93 ^(0.66***) 75*** 26*** (16.2** *) (7.95***) 6 .2W*** 1 .68*** Elder0 (liver) Somatorl^ molissima The Netherlands 196? 1968 5.5* * (63*-) 5.6** (89**) o.A*; (2.59* ) iA*; (7.2*`) California gull (abdominal fat) Western Canada L. callfornicus h " Alberta (brain) Western Canada Alberta (liver) Western Canada Alberta (ovary) Western Canada ,, Alberta 23-62.6* l8.87+= 0.81* 0.29+= 1.69: 0.79+" 2.52* . 1.22+- 138A16* 13>t+= 3.6* I .31 15-7", 5.37* 22.9; 12.56*- V g in whole egg *Rlsebrough, 1969 ++Prestt et al., 1970 "Anderson et al., 1969 Vermoor ot al., 1970 Shot and (dead or dying), respective)y Reynolds, 1971 Jensen et al., 1969 Mulhern et al., 1971 **Analysed In this laboratory an O.E.C.D. exchange sample ***this report **Koeman et al., 1970 NEV 036125 1?. Table Ilio PCB and p,p'-DDE in eggs of aquatic birds (jug/g wet weight) from the Bay of Fundy and Passamaquoddy Bay, New Brunswick (this report). D.C. Cormorant (. auritus) Herring gull (L- filmenialU) Black duck (Anas rubrlpes) Guillemot (Uria asi> Eggs Ml. Nests g No. Stn. PCB p ,p 1-DDE 11 * k 10 3 U7.0 39.3 99.1 80.8 11 F *3.52.6* 29.*ti8.5* b P 17.2iO.79 8.63+1.25 6 F 12.616.1 5.67+3.03 3P ?. 55+0.97 2.83+0.31 k 105.6 b F 9.10+0.53 1 .500.20 1 ` F' 2.02 k.3k ^Standard deviation F - Katpot Island P " Hospital Island PC 13 in .humans Fatal poisoning of more than 6kk people and fatal edema of *300.000 chicks occurred in Western Japan In 1968 due to contamination of edible rice bran oil and acidulated oil by chi orobi phenyl used as the heat-transfer medium in oil refining (Takeshita et al 19?0), Main components of the commercial PCB preparation KC~kOO (Kaneclore kOO), were 2jb,3,,*f1-, 2 , 5 , 3 '>k*-, 2y3k,k'-v and 3 ,k >31*3 '-tetrachlorobiphenyl, and 2 , 3 >kj3' jk1pontachlorobiphenyl (Sack! et al., 1971) The contaminated rice oil contained about 2300 ppm of PCB, New-born and still-born infants born from poisoned mothers-had darkish skin, possibly due to the presence of PCB which pass through the placental barrier and accumulate In the skin of the fetus (Kojima et al, 1969). A symposium report on the poisoning has boon published (inagami et al., 1969)* The components of KC-kOO with longer retention times were detected Jn sputa and fatty tissues of patients (Kojima, 1971) Serum triglyceride, cholesterol, and phospholipid concentrations were highly increased, and the lecithin-cholesterol acyl trans ferase activity was decreased. -Plasma lipoprotein lipase activity NEV 036126 ! tI DDE 3.03 O.3I `0.20 . I , 1 | 1 1 3I i I I ity ./ M .i C u a , * .,j. u J 20. was decreased in female patients (Uzawa et al., 1971; Nagai et al., )969). About h0% of the examined patients showed an elevated excretion of steroids. The 17-keto-steroids tended to increase in male and decrease in female patients (Nagai et al., 1971). Some of the patients died about one year after the accident, PCB with longer retention times, probably pentachloroand higher chlorinated biphenyls were still, after more than 8 months,found in their tissues and their presence might have been responsible for the long duration of the intoxication symptoms (Kojima et al., 1970). In Europe, PCB were detected in human milk In a concentration of 0.103 jig/ml or 3.5 Mg/g milk fat. Human fatty tissue contained 5 * 7 jug/g of PCB. Hoxaohlorobenzene was present in milk in a concen tration of 0.153 Mg/rol (5.3 fig/g fat) and the level of hexachlorobenzene in the fatty tissue was 6.3 j*g/g (Acker ot al., 1970). Hoxachlorobenzene was recently detected in human serum (Eeman et al., 1971). in the U.S.A,, two human adipose tissue samples were reported to contain 200 and 600 ppm of PCB, respectively, as determined by gas chromatography and* confirmed by mass spectro metry (Biros et al., 1970). PCB were as toxic to human cells in tissue culture as DDT (Litterst et al., 1 9 7 D . Meta bol ism. and physiological effects of halogenated Hydrocarbons Halogenated hydrocarbons, particularly chlorinated hydrocarbons, are usually metabolized in animals,by dehalogenation, Halogen atoms may be replaced either by cysteinej the N-acetyleystelne (mercaptunc acid) derivative being excreted in the urine, or by hydroxyl. In the latter case the hydroxylated derivative Is either excreted as such or in the form of its glucuronlde. If several halogens are removed from one aliphatic carbon-atom, aldehydes (such as formaldehyde in the case of bromochlororaethane) or acids (DDA from D D T ) 'are the excreted products, Bacteria and possibly mammals are able to carry out reductive dechlorination of chlorobenzenes and, probably, both insects and mammals can dehydrchlorinate chlorinated compounds. Examples of this reaction are the formation of DDE from DDT and pentachlorooyclohexene fromy-hexachlorocyclohexan (Smith, 196*0. Bacteria contain halidohydrases catalyzing the dehalogenation of halogenated fatty acids (Goldman et al., 1968). Hepatic microsomes of rabbit, rat, and guinea pig contain an enzymic system dechlorinating chloroethanes and chloropropanes (Van Dyke ot al., I 97D . This system requires NADPH and oxygen, Is JnducJble by phnobarbital and benzpyrene, and has a considerable substrate specificity. Thus 13*5, 6.0, and <0.5# of chlorine was enzymlcally removed from 1 ,1-dlchloro-, 1,1,2,2-tetrachloro-, and NV 036127 ' >i j4 .uk > 21. 1 ,2-dichloroethane, respectively. In the chloropropane series, ^0.8, 2M.6, 5*8, 5.2, 3*2, and 2.5$ of chlorine was removed from 1,1,2-trichloro-, 1 ,1-dichloro-, 1,2-dichloro-, 2-chloro-, 1-chloro-, and 2,2-dichloropropane, respectively. Very little information is available on the metabolism of PCB, Varying peak ratios observed on gas chromatograms of PCB extracted from different tissues and from animals at different trophic levels suggest that some selective metabolism of Individual chlorinated biphenyls takes place. When rabbits were fed t-chlorobiphenyl, U-(p-chlorophenyl) phenol was. Isolated from ether-soluble fraction of urine, and M-chlorobiphenylglucosiduronide from the ether-soluble fraction of urine. Analogous metabolites were found In rabbits fed biphenyl and the study indicated that ^-chlorpbiphenyl was metabolized as readily as biphenyl by the rabbit, but twice as much h-chlorobiphenyl as biphenyl was excreted in the form of glucuronic acid glycoside (Block et al., 1959). habbits can metabolize 1-chloro- and dlchloronaphthalene and in both cases, a glucosiduronic acid derivative is the ma^or metabolic product. No metabolic products of more highly chlorinated naphthalenes wore detected and It was `suggested that higher degree of chlorination may interfere with the Metabolism. There.Is a correlation botween the extent of chlorinated naphthalenes metabolism and toxicity. Naphthalene^ containing 5 or more chlorine atoms produced severe toxic symptoms (Cornish et al., 19*8). *T It is possible that the metabolism of PCB follows similar ' patterns, l.c., lower chlorinated PCB are metabolized and excreted while higher chlorinated PCB are not metabolized. PCB containing up to chlorine had estrogenic,activity os determined by the 18-hr glycogen response of the rat,uterus. MJnJmum effective dose was 8 mg. which is more, than the effective dose of p,p*-DDT, o,p'-DDE (h mg), and o,p?-DDT (0.25 mg), and equal to the activity of o,p'-DDMU. Aroclor 125^, 1260, 126.2> and 1268, Aroclor M+65 and $ b 6 0 were Inactive, Aroclor 1221, 1232, 12h2, and 12*+8 were active at 8 mg,| and Aroclor 5*^2 was active at 1 mg level; p,p*-DDB, p ^ ' - D D E . p,p*-DDMU, p,p'-DDA, and o,p*-DDD were inactive. Apparently, the reqxiirement for estrogenic activity are unsubstituted, and eJthor -OH or -OCH^ substituted p,p'-positions (BItraan et al., 1970). Administration of PCB to rabbits increases the total Jlpid, triglyceride, and cholesterol content of liver and decreases the total liver phospholipid content. The concentra tion of serum triglycerides Is abnormally Increased (Ito et al., 1V7D. The enhancement of the liver drug-metabolizing enzyme system Jn the rat by several individual chlorinated biphenyls was studied. The most potent inducer was 2 ^3f> ^ 1>5-- NEV 036120 22. pentachloroblphenyl. The effect induced with a single dose of 3 ,3* *-tetrachlorobiphenyl continued for 6 weeks (Fujita et al., 1971). Aroclor 12?U at a daily dose of 10 mg/kg (28 days) caused increased liver weights, arid carboxylesterase, aniline hydroxylase, and arainopyrine-H-demethylase activities in rabbits. No effect on body weight and on number of fetuses/litter was observed. Poses of 1 2 . 2 5 , and JO mg/kg were embryotoxlc (Vllleneuve et al., 1971). Acute oral LPJO of PCB in mice is about 2.0 g/kg. Pally dose of 0.1 g/kg administered to mice for k weeks caused a decrease in body weight and a 10-fold increase in serum triglycerides (Tanaka et al., 19&9). Polychlorinated biphenyls of the KC-hOO variety were adrainistored in a single dose to mice (2.0 mg/mouse). The concentration of PCB In the skin, one day after the ingestion was twice as high as that in the liver. Tetrachlorobiphenyls were almost completely eliminated from the tissues in 3-^ weeks but small amounts of penta- and hexachlorobiphenyls were still detectable after 9*10 weeks (Yoshiraura et al., 1971). Aroclor 1251* was found to potentiate the toxicity of carbon tetrachloride in the rat (Grant et al., 1971). Mice receiving PCB of the k8# chlorine variety for up to 26 weeks and cynomolgus monkeys (about 1 kg body wt.) receiving 6hl mg of PCB in ho days or 3^8 mg in 239 days developed enlarged livers. An increase of, smooth-surfaced membranes and reduction of rough-surfaced membranes of the endoplasmic reticulum was observed (Nishizuai, 1970). The distribution of PCB in rats dosed with PCB was determined by X-ray fluorescence analysis. PCB residues were found mainly In tho skin, and to a lesser extent in muscles, intestines, livers, pancreas, and lungs. The gas chromatographic patterns of PCB varied from tissue to tissue (Sekita et al., 1970). Pistribution, storage and excretion of PCB by Sherman rats was reported (Curley et al., 1971). Feeding PCB of the ^ Aroclor 12h2 variety depressed Vitamin A In rat liver by about (Cecil et al., 1971). PCB produced porphyria in chickens fed hOO ppm of Phenoclor DP6, Clophon A60, and Aroclor'1260 in the diet. The first 2 preparations were much more toxic than Aroclor 1260 and had higher incidence of edema and liver necrosis. PCB levels in the liver could not be correlated with the survival time and a great variation in the residue levels in liver and brain was observed (Vos et al.. 1970a). It was reported that PCB reduce estrogen concentration in ring doves (Streptopelia ri sorla) and cause thin-shelled eggs (Peakall. 1970) Howe'ver, according to a later paper, no eggshell thinning by Aroclor 1251* was found in ring doves fed 10 ppm of Aroclor 1 2 $ k in their diet for 6 months. After 35 days on the diet, the level of PCB in the eggs was h.Slil.OG ppm (Peakall, 1971). Aroclor 12jh did not induce measurable changes in egg shells or in the reproductive success of mallard ducks IAnas nlatyrhynchos) and bobwhite (Col.inus virftlnlanus) receiving 25 and Jo ppm of Aroclor in libitum diet. Lc5o!s (ppm in ad libitum diet) of Aroclor I2?2, 12m B, 1 2 51*, 1260, and 1262 for mallards were'3180, 2795, 2700, NEV 036129 At'Jm. w.iJ6u.%>Ai ii') -- * **.*. X, 23. 19 75 } and 3010, as compared to 3570, 1870, 200, and 22 for DDE, DDT, dioldrin, and endrin, respectively (Heath et al*, 1970). Similar dsta for pheasants (Phaslanus colchicus), Japanese quail (Coturnlx coturnlx)f and bobwhite were also presented. About 4-fold difference in PCB toxicity to different species was noticed. Thus for example LCJ0 of Aroclor 1254 is 1090, 605, and 2900 for pheasant, bobwhite, and Japanese quail, respectively. The toxicity of Aroclor 125*+ to Bengalese finch (Lonchura striata) is only about 1/13 of the toxicity of DDT, however, Aroclor 1254 has a much more gradual toxicity curve. Wo correlation was found between the dose and concentration of PCB in the liver. It was suggested that lethal poisoning of birds in Britain by PCB was unlikely, however, breeding failure was possible (Prestt et al., 1970). Mallard ducklings survived 10 days of feeding on a diet containing 25. 50, and 100 ppm of PCB but suffered significantly higher mortality than control animals when later infected with duck hepatitis virus (Friend et al, 1970). PCB (Clophen A 50) enhanced nocturnal activity of robins (Hrlthacus rubecula). Control birds contained from 0.0? to 0,12 v z / g PCB on fresh weight basis in breast museie. Birds fed PCB (ingested amount 55-65 Mg) contained 0,16-0,47 Mg/g in breast muscle (Ulfstrand et al., 1971). Fiddler crabs (Uca PUKilator) and pink shrimp (Pcnaeus duorarum) were shown to accumulate PCB from contaminated sediment particies (Nimmo et ai., 1971). Lethal threshold of Aroclor 1254, solubilized in water by a commercial nonionic dispersant formulation, to Gammarus oceanlcus was 0.001-0,1 mg/1 (Wildish, 1970) . Branchial necrosis was observed. Uptake of Aroclor 1254 from sea water by G,, oceanlcus was measured (Wildish et al,, 1971) . Plnfish (LaRodon rhomboldes) and spot (Leiostomus ftanthnrus) died after exposure to 5 ppb of Aroclor 1254for 14-4'iTdays, Aroclor 1254 levels in exposed fish were 46-1J2 ppm. After 04 days fish eliminated 73# on ppm.basis and 61# on total body burden basis (Hansen et al., 1971). Fatty livers were frequently observed in salmon reared in Swedish hatcheries and one-year-old salmon, average weight 18 g, had 42.2 ppm on fat basis of PCB in the liver. Eggs f-'ora salmon grown at sea contained 7 .7-9.2 ppm of PCB after stripping and 7.7-19 ppm after about 3 months incubation in fresh water. The figures for freshwater-reared females were 7*7-19 and 14-34 ppm (all on fat basis, fat content is 3*6-7.9#), respectively. High losses of eggs could be correlated with high PCB levels. The losses were due.to either unfertilized eggs or to mortality at ari early stage (Johansson et al., 1970). Aroclor 1242 In a concentration of 0.1' ppm inhibited the growth of C. closterlum (Kell et'al., 1971). NEV 036130 r *-n ;**. 2b. Latent toxicity of FCB to Chorthippus brunneus at the moult was suggested (Moriarty, 1 9 6 9 ) . ' Two compounds, 3-chloro-2-hydroxy-, and 5-chloro-2hydroxybiphenyl were repellent to Musca domestic^ (Shambaugh et al>, 1968). The latter was also effective against Totrachynup ielaxiHS. 3-chloro-2-hydroxy-5-nitrobiphenyl is an u n couplerof oxidative phosphorylation (Cheng et al., 1969). Activity of DNase I is inhibited by 2-hydroxy- and 3,l4-dlhydroxybiphenyl, while b . b -dihydroxybiphenyl has no effect (Gottosfeld et al., 1971). Fatty livers are the general symptom of exposure to vapours of chlorinated hydrocarbons such as carbon tetrachloride and 1 ,1,2.2-tetrachloroethane. The increase of the hepatic- triglyceride content has a maximum between 20 and 25 hours after exposure while the concentration of plasma triglycerides increases between 70 and 90 hours (Tomokuni, 1970), o-dichlorobenzene is more toxic to mammals than p-dichlorobenzone. Accumulation of chlorobenzene and o-dichlorobenzene but not of p-dlchlorobenzene was observed in experimental animals* Threshold concentrations for the odour and taste of water are for chlorobenzene 0*02 mg/1, for o-dichlorobenzene 0,002 and 0,0001 mg/1, and for p-dichlorobonzene 0,002 and 0,006 mg/1, respectively (Varshavskaya, 1967). Primary metabolites of p-dichlorobenscnc are 2,5-dichlorophenol and 2.5~dichloroquinol, excreted in the urine as glucuronide and ethereal sulfate conjugates. A method for their determination by gas chromatography of their trlmethylsilyl derivatives was described (McKinney et al., 1970). The toxicity of chlorobenzenes to fish increased with increasing chlorine substitution, provided the compounds were water soluble; 1 , 2 ,^-trichlorobcnzene was most toxic to Barbus, conchonlus (Toman et al., 1959). The determination of hoxachlorobenzene in grain by gas chromatography was described (Taylor et al,, 1970). Half-life of hexachlorobenzene in Japanese quail was about 3 weeks (Vos et al,, 1968). The no-effect level of hexachlorobenzene in Japanese quail ( S a i u m i x otacnJj Jap.9nlo.a) was i ppm in an &a likiiiaa for 90 days' administered diet. Slight liver damage and enlarged fecal excretion of coproporphyrin was observed in animals fed 5 ppm of hexachlorobenzene in the diet. The hexachlorobenzene levels in liver and brain were 0.79 and 0,^7 ppm, 2 ,13 end 2.32 ppm in females and males at the 1 ppm level, respectively; 6.08 and 0.99 ppm, 8.56 and l.Mf ppm at the 5 ppm level. Liver residues of hexachlorobenzene in sparrow hawk (Acciniter n isus) , buzzard (Buteo buteo) , and hooded crow (Corvus cornlx) from The Netherlands averaged **5. 5 ppm during 29?5*-i96B', and the average in I 968-I969 for the above species and Kestrel (Falco tinnunculus). marsh barrier (Circus 036131 t i- l - ^ f * * * * " * " * * * ** * m rk' 3 U ' A # .1 25. aoruginosus). barn owl (Tyto alba), long-eared owl (Aslo otus)t and crane (Orus grus) was *8.2 ppm (Vos et al., 1971/. Hexachlorobutadione caused skin irritation and hyperemia in rats (LD?0 ^.33 g/kg), and its oral LD50 was 165 mg/kg (Chernokan, 1967). Hexachlorocyclopentadiene inhibited biochemical oxidation in vater in a concentration of 0.5 mg/1, and a concentration of 0*0015 mg/1 produced detectable smell* Maximum permissible concentration of 0*001 mg/1 was suggested (Nelstein et al'* 1965). Contact eczema was caused by manipulation of timber treated with XylamonS^ a wood-protecting agent contain. ing chlorinated naphthalenes (Weichardt, 1970), Lipid-bound bromine, was detected in the adipose tissue of rats fed hrominated maize oil (BMO) At dietary levels of 0.05, 0.2, and 0.8# BMO, adipose tissue contained 60, 200, and 1050 mg/100 g lipid-bound bromine. The bromlnated compounds were not metabolized as normal lipid. Bats fed 0.8# BMO for 6 weeks contained 730 mg/100 g lipid-bound bromine in the adipose tissue* When returned to control diet, the bromine content after one week was 830 in the group fed aS libitum and 2650 in the group with restricted feeding* After two weeks the values were 7^0 and 5350. respectively (Gaunt et al., 1971a) Lipid-bound bromine was found in adipose t i s s u e l i v e r , spleen, adrenals, brain, kidneys, and lymph nodes of pigs fed BMO* None was detected in serum and urine-(Gaunt ot al*, 1971b). /. Tetra- and pentachlorodibenzofurans were detected in *'certain commercial PCB preparations (Clophen, Phenoclor). The / level of the latter was about 5 ppm in Clophen and about 20 ppm j in Phonoclor (Vos et al*, 1970b), These compounds are extremely toxic. Tri- and tetrachlorodibenzofuran causes often lethal necrosis of the llvor in rabbits after a single oral dose of 0,5-3*0 mg/kg, and a structurally similar 2,3,76-tetrachloro- dlbonzo-ip-dioxin is effective in a single oral dose of 0.05- O.I"mg/kg (Vos et al*, 1970b loc* cit*)* I&texaLnailgn ,ef .9bl9rlnated-dlbena9?PT.dioxlne. .and Chlorinated dibenzo-p-dioxins are byproducts in the preparation, hydrolytic., and pyrolytic reactions of chlorophoriols. NEV 036132 M I j . ____ --A>v>i . | * -- T*| A , . ' - - * - ....................y , ----- f ] y . j , ^ Structurally somewhat similar.to chlorinated dlbenzofurans, rnoritJonod earlier, these compounds, also known as "chick edema factor" are extremelytoxic. Recently their presence in'technical 2,4,5-T received publicity CDow Chemical Company, 1970) and a method for the determination of 2,3,7,8-tetrachlorodibenzo-p-dioxln (TCDD) in technical 2,4,5-T was developed (Storherr et al., 1971). The compound was separated by steam distillation and determined by on mJcrocoulmetric gas chromatography. Over 100 samples of 18 different pesticides derived from chlorophenols were analyzed for TCDD. Seventy-six percent of the samples contained less than 0*1 (ig /g of TCDD, 7# contained between 0.1 and 1.0 u g /g and 9# contained more than 10 u g / g (Woolson et al., 1971). Three out of 6 commercial samples of 2,4,^-trichlorophenol 5, contained TCDD, 0.07, 1.4, and 6.2 ppm, respectively. TCDD was absent from 11 examined samples of tetra- and pentachlorophenol. These samplos, however, contained hexa-, hepta-, and octachloro- ./ dlbenzo-p-dioxins, and a variety of chlorofurans and chloro- ji others. A H pentachlorophenols .examined contained from 0.17 to I 39 ppm of hexachlorodibenzo-p-dioxins, 2 pentachlorophenol 1/ preparations probably'contained a hexachlorohydroxybiphenyl. ;j I Electron-capture gas chromatography and combined gas chromato graphy - mass spectrometry were used to detect all above- mentioned compounds (Firestone et al., 1971a). TCDD was not generated in soils containing 2,^-dlchloro- and 2,4,5-trichlorophenol, but 80-85# of added TCDD was recovered from the soil after 160 days (Kearney et al., 1971). I Some of the lower chlorinated dlbenzo-p-dioxins behave 1\ simJJarly to PCB and a separation of 2,3-dichloro-, I 2.^,7-trichloro-, and 2,3,7,8-tetrachlorodibenzo-p-dioxin from PcH was described (Porter et al., 1971). Column chromatography on alumina, activated at 130C overnight was used. PCB were first eluted with 1# methylene chloride in hexane and the ehjorodibenzo-p-dioxins were eluted with 20# methylene chloride in hexane. Because of the extreme toxicity of these compounds, it is unlikely that standards will be widely available. The retention times relative to aldrin on a 6 ft - 4 mm column containing 3# 0V-I01 on 80/100 mesh Chromosorb WHP were 0.93, 1.67, and 3.0 for. 2,3-dichloro-, 2,3,7-trichloro-, and 2,3,7,8-tetrachlorodibenzo-p-dioxin, respectively. The most common standard for hexa-, hepta-, and octachlorodlbenzo-pdioxln is 1.5# reference toxic fai (available from Division of Pesticldos, Food and Drug Administration, Washington, D.C., 2020*0, The determination of hexa-, hepta-, and octachlorodibenzo-p-dioxin in tissues of chickens, fed the toxic fat, was doscribed. Over 90# of the ingested hlghor ehlorodibenzo-pdioxins was excreted. About one-half of the absorbed dioxins was present in the liver (Firestone et al., 1971b). In this f NEV 036133 i 9 s se n 0- erne DO to S ?fere jfore ind r* * 28. Table IV. Retention time and electron-capture detector response of chlorinated dibenzofurans and dibenzo p-dioxins. ggJ?.QUim Retention time ism Response/nr. Height Area h i SB-30 Column Dlchlorodlbenzofuran Trlchlorodibonzofuran Tetrachlorodibenzofurari Pentachlorodibenzofuran . Hexachlorodibenzofuran Heptachlorodibenzofuran Octachlorodibenzofuran : 2,7-dichlorodibenzo-p-dioxln trichlorodibenzo-p-dloxin 2 ,317 8-totrachlorodlbenzo-p-dioxin pontachlorodibenzo-p-dioxin hexachlorodibenzo-p-dioxin heptachlorodibenzo-p-dioxln octachlorodlbenzo-p-dioxin ' pjp'-DDE 1.65 3.35 > 6.80 ) 13.5* 27.6* 55.0* 80 1 . 9,0. 3.6*t 7.35 12.3* 22.8 25.3 *+36* 83 3.95 Octachlorodi benzofuran Hexachlorodibenzo-p-dioxln Oc tachlorod ibenzo-p-dl oxin Decflchlorobiphenyl 3* 0V-210 Column l.'+>o. 0 51.60 b .30 1.60 0.67 1.18 0.63 3.36 0.28 0.*t9 0.0*0 0.25 0.23 U.ll 6.03 16.3 0.22 0.13 0.23 1.75 ^Estimated from log(retention time) vs number of chlorine atoms plots Retention times of Aroclor 125^ peaks on the SE-30 Column were: (1.9?)> 2.90, 3.*+? rt.30), 5.1?, 6.0?, 7.20, (8.f0), (10.1); (11.5), and (1 3 .7 ) In brackets are retention times of peaks not used for quantification. NEV 036134 All evaporations were carried out In a rotatory evaporator at 20C in water-pump vacuum. Pesticide-grade hexane from Fisher Scientific Company was used as received. Pesticide-grade hexane from Matheson Coleman & Bell, supplied by Canlab, was distilled to remove a small amount of a high-boiling fraction, and pesticide-grade benzene (5 ral/1 ) was added to the distillate to obtain elution patterns identical with those of the Fisher Scientific Company pesticide-grade hexane (Zitko, 1971a). UV spectra of these 2 pesticide-grade hexanes and of pesticide-grade hexane from Halllnckrodt are presented in Appendix, An aliquot of the sample extract (2-20 ml, depending on the expected concentration of PCB and pesticides) was evaporated to a small volume (1 -1 . 5 ml) and applied to the alumina column. The solution was washed into the column with an additional 1 .5 ml of hexane and the column was washed with hexane until 20 ml of effluent was collected. The effluent was evaporated to a small volume (1 -1 . 5 ml), applied to the silica column, and washed into the.column with 1 . 5 ml of hexane. The column was washed with hexane, the first 10 ml of the effluent was collected as fraction I, the next 20 ml of the effluent as fraction IX. After collecting the fraction II, the eluting solvent was changed to a 10# solution of diethyl ether in hoxane and 10 ml of this effluent was collected as. fraction 1X1. The fractions were evaporated to dryness, the residue was dissolved in a known volume of hexane (0.2-30 m l , depending on the concentration of PCB and pesticides) and analysed by gas chromatography. In some cases a contaminant was eluted In fraction III* This Impurity appeared as a somewhat unsymmetrical tailing peak with a retention time similar to that of p,p'-DDE and could be removed by passing fraction III through the alumina coiumn, eluting with hexane and collecting 20 ml of effluent, A Packard A7901 gas chromatograph with a glass column ^(6 ft x h mm), containing SJS--30 on Chromosorb WAW 100-120 mesh, operated at 200C, was used. Carrier gas was nitrogen at a flow rate of 60 ml/mln. Injector and detector were kept at '210C, D.C. voltage in the electron-capture detector was 90V and the meter sensitivity was 3 x 10^9^, Standard solutions of Aroclor 125*+, p.p*-DDE, and 2 different mixtures of pesticides were injected daily to calibrate the detector. The concentration of the Aroclor 125** standard solution was 2.205 /xg/ml, that of the p,p*-DPE standard solution was O .96 ig/ml. Pesticide mixture M-8 contained hexachlorobenzene. aldrin, heptachlor epoxide, Pip'-DDE, p ,p 1-PDD, and p , p '-PDT in concentrations of 0.0232, 0.160, 0.302, 0,li0 1, 0.*455, and 0.*+50 ig/ml, respectively. Pesticide mixture M-9 contained lindane, heptachlor, heptachlor epoxide, dle.ldrih, p,p*-DDD, and p,p'-DPT In concentrations of 0.J55, 0.1 5 5 ,-0,302, 0.2 16 , 0.^55, and 0.^50 jzg/mly respectively. t.*--*+it*'.. mi-m. -*^ -vin*.!.,i^ ..i > ii^ :m\ J 30. To eliminate the evaporation of hexane, from the standard solutions, the solutions were kept in 6 ml hypovials closed with Viton septa (Pierce Chemical Company)* Several different volumes of the standard solutions were injected to be sure that the detector response is linear in the range used. The detector response to the chlorinated hydrocarbon pesticides, expressed as disc-integrator area per ng for all pesticides except hexachlorobenzene, where the peak height was used, is given in Table V, Table V. Electron-capture detector response to chlorinated hydrocarbon pesticides. Pesticide Hexachlorobenzene Lindane Heptachlor Heptachlor epoxide p,p'-DDE Dieldrin p,p'-DDD ' p ,p 1-DDT Kesponse/ng l*t2 * k ,3 * ? . ? k *..0.77 . 8.32 t 0.77 7.15 * 0 M 6.70 * 0,37 7.01+ i 0.32 if.96 * 0. 0k *t,56 i 0 .16 Relatlve resuonse/ng (p,p'-DDE =1.0 0 ) 1.15 1 .2 k 1.07 1.00 1.05 0.71 0.67 `standard deviation Two different electron-capture detectors were used in the present study. Detector A was in operation from January 1971 until March 1971 Detector B was used from Juno 1971* The long-term changes .in the responses of the detectors to p,p'-DDE and Aroclor 125*f are presented in Table VI. Peak area was used in the case of the detector A. peak height in the case of the detector B so that the results are not comparable in the actual response value, but they are comparable from the point of view of long-term changes. The increasing response of the detector B was not due to changes of the standard solutions on standing. Fresh standard solutions Jnjected on the 50th day gave responses of 8.59 and 3*73 for PiP'-DDE and Aroclor 125^, respectively. It is likely that the long-term trend in the detector response is caused by the conditioning of the column, as a result of which larger amounts of injected compounds reach the detector. With the electroncapture detector alone, it is impossible to measure the amount of the injected compound actually reaching the detector. NEV 036136 + b l * . . . ................... <MU> a V iA S - a .'* < ! * u i * S > . / M a i . t a t J * * 1 . ^ . 7 h. -' ; b . . 1 1 31. Table VI. Long-term changes of the electron-capture detector response* nx 1 2 3 k 5 7 8 9 10 11 12 13 1M 15 16 17 IB ` 19 20 2? 23 ?M 26 28 29 30 33 35 36 S 8 l*l 1(2 l3 '**4 '7 ^9 ;o 51 Response to p p f-DPE Detector A Detector B Response to A r o d or 125H Detector 5.77 5.42 6 .2U 6.25 5-4i 5.19 6.6l ,5.83 4.90 . 5.3$ 5.08 5.28 5.57 5-25 5.79 4.47 4.91 5.59 3.92 5.18 6.52 6*o0 5.42 5.55 5.50 5.22 >i.57 >*.60 4.37 h .29 1+.28 4.87 k .lB 6.22 5.60 5.77 5.49 5.96 6.03 6.02 6.54 5.91 6.45 6.39 7.36 6.f? . ?:8 7.75 7.56 7.44 7.46 7.6>t 7.01 . 8.14 7.77 1-59 1.H0 1.43 1.51 1.68 2.36 * 2*39 2.M2 2.69 2.35 2.68 2.60 2.77 2.78 2.78 2.82 3.04 3-19 3.03 3.26 3.47 3.47 ' J :B 3.70 .3 . 7 4 3.30 4.01 3.89 03 6I37 mem 4', t%if u ..K .^ * . frJW M . . 't --'-!-* -jw- -.-! (Table VI. Cant'd) 32. Response to p.p'-DDE Response to Aroclor 1254 Pay Detector A Detector B Detector B 55 4 .6 5 56 4.79 57 4.61 5& 5.17 61 5.04 62 4.54 63 h.81 Results of the analysis of 2 pesticide-spiked corn oil samples are presented in Table VII. The sample A was spiked in this laboratory, the sample B is an O.E.C.D, exchange sample for which the identity and concentration of chlorinated hydrocarbon pesticides was not known at the time of the analysis. Table Vll. Analysis of corn oil spiked, with chlorinated hydrocarbon pesticides. Pesticide Hoxachlorobenzene Lindane Hoptachlor Aldrl.n Heptachlor epoxide p ,p 1-DDE o ,p 1-DDT DieJdrin p ,p '-DDD p ,p'-DDT pjp'-dichlorobenzophenone Concentration, ur./ft Sample A Sample B Added Found Added Found 0.29 1.9? 1.9? 2.02 3.81 ?.09 2.72 ?. 70 ?-70 0.23 0.22 2.26 2.06 2.22 1.85 2.2*t 2.2? 3.?1 3.?9 ?.72 ?.ll 2.98 3.16 4.18 3.99 6.77 ?.?9 4.0 3.32*0.27* 1?.0 1 1 .89*1.90 . 3.? 1.06*0.1+9 5.o 3.98*0.61 7.? ?.7?*0.68 ^.? 3.83*0.97 2.0 not detected *Stnndard deviation NEV 036138 33. Average recovery of Aroclor X2$k in the analytical procedure was 10?, *?% PCB were quantified in terms of Aroclor 12?H, In some samples the areas under the second, fourth, and sixth major Aroclor 12i& peak (see the gas chromatogram in Appendix) wore used. In other samples the quantification was carried out using the total height of the first, second, fourth, fifth, and sixth major Aroclor 12JU peak. The latter method is faster and yields practically identical results as demonstrated In Table VIII, Table VIII, Comparison of two PCB quantification methods* PCB, JAg/g Sample Peak heicht Peak area Herring gull eggs Cormorant eggs Black duck eggs Herring Trout exposed to Aroclor in the lab Muscle Liver Caeca 6.92 Ul.3 5-0? 0.U3 5.50 52.5 208 7.06 W .63 O .32 6.30 57.0 228 Duplicate extracts were prepared from some samples, particularly from those with high levels of PCB and p,p*-DDE, and each extract was analyzed in duplicate. Results of single analyses are reported for other samples. The precision of the duplicate determinations was reasonably good in terms of residue analysis, the standard deviation being about 6-1?#. .Standard deviations of the egg analyses are given in Table III, additional data are presented in Table IX. NEV 036139 4 tL^M M C i h 1 3*v- Table IX. Precision of PCB, p^p'-DDE, p,p*-DDD, and p,p'-DDT determinations. Sample Herring Mean-Stand.Dev. U Jife PCB P rp'-DDE p .p 1-DDD p.p'-DDT O .kl 0.k6 0.^5 O.Hl 0. ^ 0.026 0.07 0.09 0.10 0.10 . 0.09*0.OlU 0.03 0.10 0.03 0.09 0.02 0,08 0.01 0.06 0.02 0.08*0.017 Cormorant (O.E.C.D, exchange sample) Mean-Stand.Dev. Herring gull muscle *m+*.3 373 369 1*02*32 5.01 5.12 Atlantic (whole salmon fish) smolt* abouItt II 0.M It II '11 Mean*Stand.Dev. Mean-StandDev It It 1 U .8 16,2 15.6 12.2 12.7 _ '* - trac11es - II II - II lU.3i l .76 - 2.13 1.99 -- traces -m 0.U6 0.U9 ' 0.39 0.37 0.72 0.67 0.79 0.69 1.10 1.22 1.^2 1.21 o.U2*o.o?7 0.72*0.053 1.214*0.13 0.68 1.0U 0.12 0.18 0.06 0.07 O.Ul 0.38 0.39 0.35 0.39 0,33 0.63*0.30 0.26*0.13 0.20*0.19 ^Exposed to technical DDT; different fishes were used to obtain duplicate extracts. NEV 036140 yi'ilimw^iv -ivi.'k^el.i-.u.mA^Ll, \ t v>: . , * 35. ; CONCLUSIONS j * j A variety of halogenated compounds is used in j Jndustry and incorporated into numerous products* These j ' compounds enter the environment and may be accumulated in { the biomass as already demonstrated in the case of PCB, j hexachlorobonzene, byproducts from vinylchloride production, j and PCT. PCB and hexachlorobonzene were detected quite easily < ^ since they accompany chlorinated hydrocarbon pesticides during the analysis* Modified conditions had to be used to detect low-molecular weight chlorinated hydrocarbons and PCT, It is very;likely.thatoother halogenated hydrocarbons will also be detected" in the environment as soon as suitable analytical* methods' become available, ' Some of the compounds may already now bo seen as unidentified peaks in the routine determination of chlorinated hydrocarbon pesticides. Very little is known about the fate and toxicological J significance of industrial halogenated hydrocarbons in the environment. Levels of hexachlorobonzene in livers of some spocles of birds in The Netherlands are well above those found in birds fed on a diet containing hexachlorobenzene at the no-effect level in tho laboratory. No dose-level-effect j relationship has been established for PCB. The rate of accumulation of PCB and hexachlorobenzene j in the environment- is not Jtnown, since the levels have not been j monitored for an extended period of time. It is possible \ that a steady-state situation exists in some areas. The world j consumption of industrial halogenated hydrocarbons should be determined. This together tfith some toxicity data already j available and some extrapolations would determine research \ priorities. Until more refined data on acceptable levels of ] haJogcnated hydrocarbons in the environment become available, thoso compounds should be considered potentially harmful and their discharge into the environment should be limited as much as possible. I In studies of sublethal effects of PCB on fish and i other aquatic animals, great care should be taken to secure ! uncontaminated controls, mainly by using a PCB-free diet. i ; 1 NEV 036141 i s i l .Mil*--*f- - -- w * !.. I . ^ _ ...^ - . . . _ ._ ACKNOWLEDGMENTS 36 Wo thank our colleagues at this Station Tor providing tho samples. Writing of the report wa? greatly facilitated by the efficient assistance of Mrs, Madelyri M, Irwin in the documentation of the literature and by the skillful help of Miss M, Beryl Stinson and Mis.i Linda L. Cunningham at the library of this Station. We thark Dr. D. Firestone, Acting Head, Food Alterations Section, Biochemistry Branch, Division of Chemistry and Physics, Food and Drug Administration, Department of Health, Education, and Welfare, Washington, D.C., for samples of chlorinated dibenzofurans and dibenzo-p-dioxins. We thank Dr. D. J. Wildish for helpful comments on the manuscript, Mrs. Madelyn M, Irwin typed the manuscript, Messrs, P,W,G, McMullon and F, B. Cunningham prepared the figures. REFERENCES ' Acker, L . , and E. Schulte. 1970. Na turwl ssense ha ffen 57. **97. Ahllng, B . , and S, Jonsen. 1970. Anal. Chem. , ll*83. Anderson, D. W, , J. J, Hickey, R, VI. Risebrough, D. F. Hughes, and R. E, Christensen, 1969- Can, Field Naturalist 3, 91 Anon. Halowax Chlorinated Naphthalene Oils and Waxlike Solids, Koppers C o., Inc. Anon,, 1969. Hooker Industrial Chemicals, Bull. 100. Anon. 1971* Chem. & Engng. News Jt2(7), 6?, Armour, J. A,, and J. A. Burke, 1970. J. Assoc, Offic. Anal, Chemists 3, 761. Armour, J. A,, and J, A. Burke. 1971 J. 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Naturwissen schaften 8, 276. Zitko, V. J970. Bull. Environ. Contam. Toxicol,, 5, 279# ZJtko, V. 1971a. J. C h r o m a t o g r . 2 W * . Zitko, V, 1971b. Bull. Environ. Contam, Toxicol., (5). Zitko, V., 0, Hutzinger, and S. Safe, 1971a. Bull. Environ, Contam. Toxicol,, 6, l60. ZJtko, V,, 0. Hutzinger, W. D. Jamieson, and P. M* K. Choi. % . 1971b. In preparation. t11 . , 036151 UEV -~.~ {-.Cffc-J......... .. J . - . . I H6. addkndum Additional information was obtained while this report was in typing, PCB were characterized by gas chromatography on SK-30 support-coated open capillary columns. ^Cl-labelled Aroclors 12h B and 1251* were prepared by neutron irradiation (Stalling et al,, 1971)i and the preparation of 3n-iabelled PCJ3 by-selective labelling and nonselective chlorination in the gas phase over ferric chloride was described (Igekami et al., 1971). In addition to chlorinated paraffins and PCB, sheet materials may be fire proofed by pentabromotoluene, tetrabromobenzene, trichlorotribromobenzene, decabromobiphenyl, chlorinated rubber, bromineted soybean oil, and perchloropentacyclodecane, Chlorinated paraffins are used for cotton, paper, and unbleached kraft based boxboard and corrugated medium. Adhesive compositions for laminates may, in addition to chlorinated paraffins, contain ?CB of the and 60jS varieties (Thiery, 1971). Flame retardant compositions for polyolefins, poly (oxymethylene), nylon, and cellulose acetate are based on chlbrinated hydrocarbons with boiling point above 200C, containing more than h0% chlorine, and phbsphlnic .acid derivaties. Hydro carbons frequently mentioned ih this report, and hexachlorobenzcne, 2,3 >9 ,6-tetrachloro-p-xylene, and <x,a-dichloro- m -xyleno con be used (Cannelongo, 1971). Chlorinated paraffins Cereclor U2 nnd Cereclor 70 are used in high-build marine primers (Bowerman, 1971). Fire retardants based on polyhalo polyhydro polycyclic dicarboxylic acid Imides were patented (Cyba, 1971). TCDD, administered ornlly to pregnant rats on days 6-15 of gestation, had no effect on foetuses pt the 0,03 Mg/kg day level and foetal mortalities were observed at and above 0,125 jig/kg day. Decrease of maternal weight gain started at 0,5 pg/kg day and a severe toxicity was evident at 8,0 tg/kg day (Sparschu et al,, 1971). Iloxachlorobenzene is neurotoxic in male rats (Lehotzky et al., 1971). Current U.B, maximum acceptable levels of PCB in eggs, and ready-to-eat poultry and pork are 0,5 and 5 ppm, respectively (Anon., 197Ja,b), 1CB contaminated fish meal, feed, eggs, and chickens wore very recently found In the U,S.A. Fish.meal imported from Peru contained 6.7-7*3 ppm of PCB, Heat scrap meal at an Illinois plant contaJned PCB at 1 ppm. The range of PCB found in total edible ticsu Jr, 0-5.3 ppm In hens, 0-0.7 ppm in broilers, and 0-11,8 ppm in turkoyr; (Anon., 1971b). It is not clear at the moment how much of. thJr, contamination is causod by industrial accidents (PCB leakage NEV 036152 s* * ! ,: m u , V. ^ ,.. *7. from boat-transfer systems) and what is the contribution of the environmental levels of FCB. The concentration of FCB and chlorinated hydrocarbon pesticides was determined in a few additional samples and the results are presented in Table X, Table X, Additional data on PCB and chlorinated hydrocarbon pesticides in aquatic animals. Species Location FCB p ,p 1-DDE Mg/g hg/g Other Basking shark Eastern Canada Cotorbir^us^gjix^inus (Gunnerus) (liver) (muscle) (liver) White sinhnnrrKk Leeoonnaarr<)gvvinlilee, wN..bB.. n MlfiggOfc S t e f 8> (" (liver) " pect. fin) 0.06 1.37 0.0 7 0.77 o .6 k 0.77 218 Biuefin tuna Eastern Canada IbBJmus thynm^s (rnusclc) 1.5H Volsclla demi ssa ilchi estuary, N.B. 0.09 Clam Ba Mya aronurla . B. 0.09 traces h 0.57 oM 335 DL'T 63, DDD *43 0.15 0.01 traces Kuforcnces Anon. 1971a. Chem. & Engne News i+2(3*+) 30. Anon. 197)6. Food Chemical News 13(23), 3 iJoworuian, D. F. 1971* Faint Manufacture 31, 20. 25CanncloriGo, J. F. 1971. U.S. 3,582,510 (Cl. 260-28.5; C 08fg) J June; CA , 50057 (197D. Cyt'.'i, II. A. 1971. U.S. 3,57^,230 (Cl. 260-326; C 07d) 6 Apr.; CA 25, 500142 (1971). < NfcV 0 3 6 1 5 3 r-u fli tvteufc tx k * M lt *A v^ k /v*. l u i v j ^ X i '.* ^ vJ` ^ a <A > W. ^ Igekami, S,, K, Kawamoto, Y. Kaslda, S, Akaboshi, K, Enogaki, and S. Baba, 1971* Radioisotopes 0, 65. Lehotzky, K . , I. Desi, and S. Bordas. 197^- Egeszegtudomany 15, 52; CA 25, W 3 0 (1971). Sparschu, G, L. , F, L, Dann, and V, K, Rowe. 1971. Fd. Cosmet. Toxicol., 2, ^05. Stalling, D. L . , and J. N. Huckins. 1971. J. Assoc. Offic. Anal. Chemists 4, 801. Thiery, F. 1971. Papeterie 21, ***+0.