Document 5LadzxpbDyv7abQ5bwgRX30Je

Current Progress in the Determination of the Polychlorinated Biphenyls * ^ - -. * K. \V. His>.nuovcn, P. Rkiciif., tui H. S. Olcott InMitotr ofMnrinr Hrintirfrs '. Dejtartiiirnl of Sntrilinnnl Sciences , ; , '_ . ' University of California ' - Berkeley, Californio . .. . University of California, Berkeley,'California : The waste produces of global technological society * W Inevitably becoma components of the environment. Many of ` ` these waste materials have the capacity to inflict permanent , . changes upon the ecosystem. Among tho synthetic pollutants which are accumulated by wildlife the chlorinated hydrocarbons !. ... of blocido origin appear to be the most abundant. Residue ' - . levels in marine fish and birds now frequently surpass those ; ; ' jrecorded in terrestrial and fresh-water species (1,2,3). In - . ' California, the concentrations of chlorinated hydrocarbons Vv : '' ' ' ..' in petrels, pelagic birds which do not approach land except ' `; to breed on off-shore islands, are many times higher than - - ' those recorded in species inhabiting agricultural and urban ` \ . areas (1). Other classes of environmental pollutants which might be similarly accumulated by organisms and similarly j ' ` ' dispersed around the world could be expected to share some - ' ; ' ; ;or all of the following properties of the biocide derivatives ' " . . . - Which have contributed to their unexpected-distribution, in'" ... V . .. _ _. ; . - - j ' -.the global ecosystem. -V. ' , ;' ' . . ' . 1) High world production. U. S. production of DDT in 1965 . '- . ' \ ; kallrtin f E/liirunrtwni.l Cpniimin.iivn & Toxirulcfy. ' ' - . Vl 4. N, 4 pulilnird liy Spfi|wT*Vrri,fl Nm Yiwk lw. < ; ., ' mmtrn tim+nwtmuwn i| iP>|wn tff m iy / i.wu^i .i i.f.i 1 i / .V *- i i DSW 552344 i STLCOPCB4090761 r~. wa 140 million pounds (4). World yearly production is therefor* v In the order of 10^^ grama, only five orders of magnitude leas ; than the amount of carbon fixed by plants into organic matter (5). - Other pollutants which arc present- in ecologically significant amounts in the global ecosystem must, like p,p'-DDE, therefore derive from primary materials that are produced in comparable amounts. ' 2) Chemical stability, p,p'-DDE, which nay be tha rapot abundant of the synthetic pollutnntB in the environment, is comparatively resistant to degradation by the usual detoxification mechanisms of vertebrates, to microbial action, and to non-biological breakdown in tha environment. Its half-life, therefore, would appear to ba greator than ten years. . . .. - ' 3) Insolubility In water. Water-soluble waste products will bo diluted to a greater extent in.the sea and may or may not be * subsequently accumulated by organisms. Tha non-polar nature of P*P'"DDE and"'fho other chlorinated hydrocarbons explains why ' - they are concentrated in fat tissue and why they are present in - greatest amounts in- the terminal carnivores of food' chains. ` 4) Mobility and aerial dispersal. Even though vapor pressures of potential pollutants might ba very low, the amounts entering ; .the atmosphere could become ecologically significant over periods ' *. ' . *r of time, especially when rates of vaporization are greatly ' ' Increased by codistlllatlon with water (6). Incineration of ' ` i 3 pm ^ ii uww.w waste mat vaporizac can be fu fallout. Amor. the arocl Their use resins dc and resio Unid have long and birds polychlor evidence. 1) Mass si compounds with a cor chlorine-t as biocidi 2) Retcnti of peaks 1 cannot be have rater, are identi DSW 552345 STLCOPCB4090762 o K 1< I A* .* *.!* *) ^,waste materials would also significantly increase the rates of ' j.vaporisation of none compounds. The global distribution of DDT "Can be fully explained only by aerial dispersal with subsequent o;fallout, in conjunction with transport by water (2). .;u'.v .-.l..Anong industrial products which meet these criteria are ^(t;hc aroclors, which are mixtures of chlorinated polyphcnyls. tllThcir usefulness in the manufacture of many plaotics, paincs and ,rpBoinp decivos to a largo part from thoir chemical stability ijond.,resistance to degradation*. ...... . , . of , ..Unidentified peaks produced by halogen-containing compounds ;.i.bavo. long been evident in gas chromatograms of extracts of fish o;ond birds... The identification of cost of.these compounds as . polychlorinated biphenyls (?CB) is now based upon the following /. evidence. _\ \ . .: 1) Hans spectromotrlc analysts in Sweden. Hass epectra of - compounds present in extracco of fish and.sea birds obtained with a combined gas chromatograph-mass spectrometer showed that chlorine-containing compounds which could not be identified ' _,.aa biocides were polychlorinated biphenyls (7). . 2) Retention times In gns chronntographlc nnalv3ln. The majority . of peaks in chromatograms of extracts of fish and birds which - cannot be identified as biocides that emerge after p,p'-DDE , ", have retention times on both polar and non-polar columns which ' \ \ are identical with those of the principal PCB compounds The J rr'n.v* n.^imny pw,ww ,52^46 STLCOPCB4090763  .:VK.O.-.v-. . 'I* 0 najor peaks produco'a characteristic pattern on each kind of column. The same patterns of peaks aro evident In extracts of wildlife from around the world (1,2,8,9). - '* 3) Chlorine content. Mlcrocoulometric analysis shows that tho compounds'identified as PCB contain halogens. . ' 4) S.ipon 1 f lent ion and nitration. Unlike the peaks of toxaphene (10K p,p'-DDT, p.p'-DDT, and p,p'-DDD, the PCB peaks arc not a removed or displaced by dchydrohalogcnation with alcoholic KOH. Vigorous nitration carried out at room temperature with fuming nitri'i acid (11) removes the chromatographic peaks of both PCB and the DDT compounds, but docs not destroy the toxaphene peaks. At 100* the DDT compounds arc tetranitrated by this procedure - (12). Tho nitrated derivatives would not pass through the ' commonly used CI.C columns. A milder nitration process, carried out at 0* with nitric acid rather, than fuming nitric acid, .. U., -- destroys the-DDT, but not the PCB peaks. . . , ' 5) Distribution In the global environment. Concentrations of PCB in fish and birds are highest In such' industrial outfalls : ' as San Francisco Bay and San Diego Bay and concentration gradients apparently exist from these areas to regions more remote (1,2). ' A consistency in tho ratios of p,p'-DDE to PCB among the birds of a given region suggested the existence of regional fallout *0 patterns (1). PCB has been found in all sea birds from tho *\ \' Pacific Ocean so far analysed, but they could not be detected ' / in pcngul / compounds The chromatog with thos DC-200: _ QF-li . The columi Chromooort of 195*. underline! on DC-200. peak emert On QF-1 cc with o,p'with tha c X lntcrferer shoulder c V As a of tha v&l .recent lit \ . reported i TT DS\N 552347 STLCOPCB4090764 / ; : , . i in penguin eggs from thc/Antarctle which did contain DDT coxpounda (1). Tha retention tinea relative to p.p'-DDE, of pcaka in . chromatograms of extracts of fish and birds which are identical . with those of PCB compounds in the commercial preparations arc; . DC-2001 1.25. 1.48. 1.75. 2.05, 2.41, 2.50, 2.90, 3.41, 3.88, . 5.53; dleldrin: 1.00; p.p'-DDDs 1.27; p,p'-DDT: 1.68 ' -.* QF-li 1.10, 1.33. 1.40. 1.65, 1.72. 2.14, 2.59, 3.23, 3.88, . - 4.84; dicldrin: 1.49; p.p'-DDD: 1.75; p.p'-DDT: 1.91 * The columns employed were 10Z DC-200 and 3Z QF-1, both on , . Chroaosorb W, 80-100 nesh and were maintained at a temperature ' . of 195*. The retention times of the; three principal peaks are .. - ' . underlined. The first of these, with a retention.time of 1.25 . ".on DC-200 columns emerges with p,p'-DDD. Another major PCB . . >' peak emerges a lightly later than p,p'-DDT on DC-200 columns. .. On QF-1 columns, Cho compound with retention time 1.33 emerges : . ' , with o,p'-DDT and the compound with retention time 1.75 Interferes . ^with the determination of p,p'-DDD. On this column there is no . . interference with p.p'-DDT. Dieldrin appears as n trailing .. ; shoulder on the peak with retention time 1.40. . V. ' As a result of this interference, it appears that many , ; . . of the values of p,p'-DDD, p.p'-DDT and o,p'-DDT reported in : .recent' literature arc erroneous. Tue DDE values originally ` , reported la Pacific sea birds in a paper from this laboratory . . \ STLCOPCB4090765 ttU)UiuCt* /'* iliiO '*~Y '- .U af t. (13) were coo high'because of'PCB interference and have subsequently been corrected (1)*' p,p'-DDE is,$he roost abundant i.i .... ;... . of the DDT compounds in the environment and there is no significant FOB interference in the determination of p,p'-DDE. Consequently the total DDT residues reported in the past, before the extent of PCB interference was known, would not be greatly changed after correction for this interference. o,p'-DDT, which has been shown to be an active estrogen (14,15) is present in ;/ very low amounts in the global environment, although o,p'-DDE is comparatively sore abundant (1,3). - .. When both DDT and PCB peaks ere present in chromatograms. . " the clua to the relative amount of PCB coses from the height . ` of those PCB peaks which do not interfere with any of the DDT : compounds. With the exception noted below, the three major *~^ ` peaks usually hava approximately the some height on chromatograms obtained with an electron capture detector. If the peak with ' retention time 1.48 on DC-200 columns is approximately as high as tho "ODD" and "DDT" peaks, no or very small amounts of p,p'-DDD and p,p'-DDT arc present. When one or both of these peaks Is relatively higher than the PCB peak with retention time 1.48, the amount of p,p'-DDD and p,p'-DDT can be estimated from ' the changes in peak height with saponification. The saponifi- -\ * cation procedure is rapid, and alnce only -a -relative change in peak heights is determined, it need not be quantitative. An . - / ............,............... \. - - - - - \\* . m -' * ' ' eliquot of. : in ethyl a; i j. aqueous NsC i j the hexane } have been i i contrlbutlo | can thereby PCB ha hydroxylasc partially i observed A: of comparab - Since individual future, the (10), also present be . used (1.2) microcoulom PCB compounfish and bi by the coma' nr WV'U'JI Ullfll qSVJ 5523^9 STLCOPCB4090766 aliquot of the extract is refluxed for approximately 5 .Minutes // In ethyl alcohol with 5Z K01I, to which hexane and concentrated aqueoua NaCl aolutlon are added in turn. Chromatography of .I the hoxene/laycr shows that the p.p'-DDD and p.p'-DDT peaks have been displaced, but the PCB peaks are unchanged. The contribution of PCB to the peaks of the original chromatograms can thereby be estimated, provided the detector response is linear. ^. PCB hao; been shown to be a powerful inducer of steroid ^hydroxylaoco in birds (1) and, with p,p'-DDE; may therefore be ' : '.partially responsible for the aberrant calcium physiology _.observed in spcclca of raptorial and fish-eating birds which ^accumulate chlorinate^ hydrocarbons .(16,17)... It.is probable ' . ..that the PCB in human food supplies would Induce the .synthesis ' . ' .jOf comparabla enzymes in man... ... . . ...-, .. . ... ... .. ; ) - .n.v-. . S inc---e----i-t *is highly*unlike.l.y.t.h.at primary standards of '. ., . ; individual PCB compounds will be available in the foreseeable .' Vvt. .............................. ...- ' ^ futuro, the^ quantification procedures, like those for toxaphene / ' (10), also, a mixture of.polychlorinated compounds, must for the U#.pS*reCs' -eknt be.*a.-ppr4ox-i.m....a..t.e......... T* he.....q...uant'ific*ation procedure we have . # " ^used (If?)., is based primarily, upon the. determination with a microcoulomctric dettector of. the total chlorine content of the ' PCB compounds emerging after p,p'-DDE. The. chromatograms of .; *. ' OfUiskh: and Wblwirnde..x' trac...t.s.....m...o..s..t*,'clo*sely.*r'essemble' * those.-p*roduced -. by the commercial,.aroclor.which has an average chlorine content' ) 'as the "DDD" and "DDV' peaks,, no or very'small ;;i.-.oent^.n: . * P.V-DDD and P, t>'-0DT.-.a\je `present. one or both ' ` peaks is relatively higkc\ tnar. `j}} pea.c with v > . .- . r 1.43, the amount of p.p'-o^i '.nd p.p'-Sk. one oo "S--.:--........... W"* l the changes in peak height vttn -v.'. >,1 ! 'H m IH'III uni. .mull,n.i ; \''v; V--1 f*v *: .;v. ' *\ . i *. '"-^1 . ,.. : .-. ' 'v i. ' ' \ DSW 552350 STLCOPCB4090767 /*~N o. v ii ' 4. <\* i. I ,'bf`542.'`PCB concentrations in fioh and birds were therefore .* J derived iron'the chlorine values by assuming that the chlorine , .WonVent was also1 542.' 1 This procedure would occasionally include .. other chlorinated pollutants with the PCB. It Bay, however, be more ceaningful to measure total organic chlorine (18) of non-biocide origin in tissues of wildlife than parts per ! Billion concentrations of individual compounds. This might i be a bettor index of physiological effects such as enzymo Induction. , Since we do not yet have in our laboratory the permanent . use of a nicrocoulomccric detector, wo have had to depend upon electron-capture for routine quantification of PCB. Each PCB compound was initially assumed to produce the same peak height, with the electron capture detector, as an equivalent amount of . - p,p'-DDE. Amounts of sample injected into the gas chromatograph .7 . were adjusted in order to fall within the linear response . range of the electron capturo detector. Tlsc amount of total .. PCB considered as p,p'-DDE was then multiplied by a factor . ^ . which would yield a value equal to that obtained with a micro- _ coulometric.detector. This procedure was periodically checked by electron capture determination of standard PCB mixtures. A1chough this method yields approximate results, it docs .. \ - permit accurate determinations of relative concentrations. It \ - ' . \ . " -has shown.that some species of birds, notably certain raptorial . species, of PCB 0 and conce also shov concentre The and 1.33 chromatop Bay and 1 is presen the Gulf aerial fa * Californi tested tfc selective in the at evaporate dishes co / rthcn irra gormicida of peaks essential height of was irrad -***" * V*n ^ V..' | SO w> f' ' *< Ill , i n " T^i i n , i STLCOPCB4090768 .K -.4 species, shearwaters and petrels, accumulate high concentrations of PCS (1). Terminal carnivores of food chains may accumulate and concentrate the PCB present In prey species (1). It has also - -sh. ow.`n\.r. that.....r...e......g.....i..o.....n. al fallout - patterns exist ** (1), with highest concentrations in Industrial regions. . ' ; The PCB compound with retention time 1.25 on DC-200 columns ' and 1.33 on QP7I columns produces one of the prominent peaks in chromatograms of extracts of fish and birds from San Francisco Bay and in the commercial mixtures, yet it is not present, or is present in relatively low amounts, in extracts of birds from the Culf of California. PCB in this area must derive from . ' aerial fallout, since there are few Industrial areas in Baja California or along the coast of western Mexico. We therefore tested the possibility that this compound might have been ` ~\ selectively degraded by ultraviolet light during Its transport in the atmosphere. Standard PCB solutions in hexane wore 1 evaporated.to dryness in Petri dishes, or were applied to Petri dishes containing either water of 3Z NaCl solution. They were - ..................................-............................................................................ ... - ... . then irradiated at a distance.of. 4 inches with a 15-watt ' germicidal lamp emitting at 2537. A. In each ease the pattern **/.'- *' ' ..... . '.................... ' ' ' " / of., peaks on the chromatograms of the irradiated PCB was; ' `' ; essentially unchanged, except for a relativejreduction /in the \. * height of this. peak.. The ..effect, was. more pronounced wi>en PCB COt< a i. a* a< ' *' t 1 ` v*. -t v 'I ** ' . yi>ays irradii*a'<tr.edC,oopnevt>h. e .aq. ueous, surfaces .-'i - - ..... - J* - * * Although thistletV.od yieia.t 1 / permit accurate doceXi.'.Atioaa e.f relative. has shown that some species of ' i '.' ' . .'' : ' \ . ' \ . \ '' r.ocabiv . .. :. / " .; _ ' r 1 55^52 qSNN STLCOPCB4090769 >, . V Acknowledgements ' * - - Vo thank tho Monsanto ChenicAl Company for supplying samples of tho Aroclor3. Tho National Scienco Foundation (G36362) provided financial support. Roforoncos 1. R. W. RISEBROUGH, D. B. PEAKALL, S. C. KERMAN, M. N. KIRVEN and P. RIECHE, Naturo 220, 1098 (1968). . * -" \ 2. R. W. RISEBROUGH, Chemical Fallout, Firot Rochoster Conforonco on Toxicity, in tho proas. \ 3. R. V. RISEERCUGH, D. B. MS21ZEL, D. J. MARTIN and H. S. OLCOTT, in preparation. li. Tho Pocticido Rovlow, United States Dopartnont of Agriculture . .(1966).. . ... ... ' 5. E. S. NIELSON, Ann. Rov. Plant Physiol. 11, 3^1 (I960). . ' 6. F. ACREE, Jr., M. BEROZA, and H. C. BO".vMAN, Agric. Food Chem. 11, 270 (1963). % t * ., ** ' ' i 7. C. WIDMARK, J. Ass. Off. Anal. Chom. $0, 1069 (1967). ' ' 8. D. C. HOLMES, J. H. SIMMONS, and J.O'G. TATTON, Naturo 216, ' ; 227 (1967). ' ' ` .. * '* .. '. 9. A. V. HOLDEN and.X. MARSDEN, Naturo 216, 1271 (1967). J ' 'I 10. T. E. ARCHER and D. 0. CROSBY, Pull. Environ. Cont. Toxicol. 1, 70 (1966). . :. ' 11. F. ERRO, A. BEVENUK, and H. BECKMAN, Bull. Environ. Cont. Toxicol. 2, 372 (1967). .. . . ` 12. H. S. SCHECTER and H. L? HALLER, J. Am. Chem. Soc. 66, 2129 (19UU). . 13. R. E. RISEBROUGH. D. B. MENZEL, D. J. MARTIN, and H. S. OLCOTT, Naturo 216, 509 (1967). .. U. R. M. VELCH, V. LEVIN, and A. H. COIJNEY, Toxicol.' Appl. . Pharmacol., in the proso. 15. J. BITKAN, H. C. CECIL, S. J. HARRIS, and G. F. FRIES, Scienco 162, 371 (19,68). 16. D. A. RATCLIFFE, Naturo 215, 208 (1967). _ 17. J. J. HICKEY and D. V. ANDERSON, Science 162, 271 (1968). 18. F. A. GUNTHER and J. H. BARKLEY-, Bull. Environ. Cont. Toxicol. 1, 39 (1966). \ - ' \ 301 ' . \ / M pn VT / *' Expa i; for Or: and the C E In 1 utility o compounds to signal, for facil. fractions^ 0.17 pg. < visually < used at t: minimum d< ! Bul^tin of Environ VoL 4, N*. 4, 1969.* qSW 552353 i i \ i STLCOPCB4090770