Document 10MzYvwGJKeZyGoDDNdEabJMd

Current. Progress in llie Determination of the Polychlorinated Biphenyls *7 H. W. HiM-nmuxii,. P. Rkiciik, arid It. S. Olcott Inihhtlr | Mnrinr Hrtnurm . thrfHtrinifnl >/ Sritntrt , . . Unitvr.tiiy rtf Cnli/ornin ' Urrkelry, Cnlijarni* .. Unilvriity of Cnlilornin, Brrkt`lry,<Cnlilnrnui ` Tho waste products of * global technological society Inevitably become component* of the environment. Hony of ` these waste materials have the capacity to inflict permanent changoa upon the ecosystem. Among tho aynthotic pollutants *1 ' ' which are accumulated by wildlife the chlorinated hydrocarbons < 'i . of biocide origin appear to bo tho most Abundant. Residua levols in marine fish and birds now frequently surpass thoso ' jrecorded in terrestrial and freah-water species (1,2,3). In California, the concentrations of chlorinated hydrocarbons vJ ' in petrols, pelagic birdo which do not approach land except . to breed on off~ahoro islands, aro many times higher chan ' those recorded in species inhabiting agricultural and urban . . areas (1). Other classos of environmental pollutants which . might bo similarly accumulated by organisms and similarly ' dispersed around the world could be oxpccted to share some ' : 1 . ' . '' ' ' ''i ^ or ell of the following properties of the biocide derivatives " Which have contributed to their unexpected distribution, in ' * : , tha global ecosystem.- . 1) ntgh world production. U. S. production of DDT In 1965 . . . ":' / \ : 192 /- A\ \ . '!.:' V*1 4. Nm 4. IMS, pvlil*WS )>/SptinjnT>VrrU* Nr* Ywk lan, ' . -. ,, 1 / -/ V. / t - 7 DSW 033697 va* 1A0 million pounds (l>). World yearly production 1b therefor* In tho order of 10^ grans, only fiva ordcro of magnitude less ; than the aaounc of carbon fixed by plants into organic matter (S). Ocher pollutants which are 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 oay bo tho opot abundant of the synthetic pollutants in the environment, is comparatively resistanc to degradation by the usual detoxification mcchanlemo of vertebrates, to microbial action, and to non-blological I. breakdown in the environment. Its half-life, therefore, would - appear to ba greater than ten ycaro. . - 3) Insolubility In water. Water-soluble waste products will bo diluted to a greater extont in,the sea and may or uay not be ` subsequently accumulated by organisms. Tha non-polsr nature of p,p'-DDE and tho 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. Mobility and ncrlnl dispersal. Even though vapor pressuras of potential pollutants might be very low, the omounts entering " .tha atmoaphera could become ecologically significant over periods <. a. of time, especially when rates of vaporization era greatly , ' Increased by codistillation with water (6).' Incineration of ' waote ma> vaporiiat can be ft fallout. Amor the arocl Their uac resins dc and reels Unid have long and birds polychlor evidence. 1) Mass hi compounds with a cor chlorine-< ss blocld' 2) Recent! of peaks l cannot be have rater, era identl u'l* to ** I / . . . . ... k^ Vv \ t ,4* . 4 C. #*)*. >M|. , % * ,,woote materials would also significantly incrctao the rates of ,,y ,.valorization of sotao compounds. The global distribution of DOT -esn be fully explained only by serial dispersal with subsequent ^fallout, in conjunction with transport by water (2). Among industrial products which meet these criteria are <<(thc nroclors, which are mixtures of chlorinated polyphcnyla. CllThclr uaofulnpss in the manufacture of many plantics, paints and />rjcasinp docivos to a largo part from thoir chemical stability ijsnd. resistance to degradation*. ^... .. . . , of . ..Unidentified peaks produced by halogen-containing compounds ^i.havo.long been evident in gas chromatograms of extracts of fish o;cnd blrdo. , Tha identification.of most of . these compounds ss . ; polychlorinated biphenyls (PCS) is now based upon the following /. evidence. \ 1) H-ins spcctromccric analysis In Sweden. Mass i.pectra of o compounds present in extracco of fish and.sea birds obtained with combined gas chroaatograph-aase spectrometer showed chat chlorine-containing compounds which could not be identified ' ....as biocides were polychlorinated biphenyls (7). 2) Retention times In gno chronntogrnphic analvsli. The majority of peaks in chromatograms of extracts of fish sad birds which - cannot be identified a8 biocides that emerge after p,p'~DDE . have retention times on both polar and non-polar columns which ' - are identical with thoaa of the principal PCS compoundo. The . ; . . . . . ; '\ * -A 'y. V.. 7 , w : % * J DSW 033699 n major peaks produce 'a characteristic pattern on each kind of V/ column. The sane patterns of peaks era evident In extracts of wildlife frois around cho world (1,2,8,9). - 3) Chlorine content. Mlcrocoulomctrie analysis shows that the compounds*identified as PCB contain halogens. - ` 4) Saponification and nitration. Unlike the peaks of toxaphene (10)* p.p'-DDT, p,p'-DDT, and p,p'-DDD, the PCB peaks arc not removed or displaced by dchydrohalogonatlon with alcoholic KOH, Vlgorbuo nitration carried out at room temperature with fuming nitri'd add (11) removes the chromatographic peaks of both PCB and the DDT compounds, but does not destroy the toxaphene peaks. At 100* the DDT compounds arc tetranltratcd by this procedure 1 (12). The 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, .. -- destroys the-DDT, but not the TCB peaks. . . ............. ` 5) Distribution In the global environment. Concentrations of * PCB in fish and birds arc hlghcot in such industrial outfallo 1 as Sen Frencisco Bay and San Diego Bay and concentration grodlenta 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 roglonal fallout *a patterns (1). PCB has been found in all sea birds from tho \i ' . Pacific Ocean so far anolyaod, but they could not be detected ` * in pcngul compounds The chrooatog with thoe DC-200: QP-lt The colurc Chromooor of 195*. underline on DC-200 peak emer z' On QF-1 c with o,p' with the tN Interfare shoulder As a of the val .recant lie \ reported 1 i uiyn*,'u i"5i'..srs"..Mi>n' "/ OSw 033700 STLCOPCB4017662 / In penguin eggs from Che Antarctic which did contain DOT compound* (1). . The retention times relative to p,p'-DDE, of peaks in . chromatograms of extracts of fish and birds which ora Identical , with those of FOB compounda In the commercial prcparatlono are; DC-200 1.25, 1.48. 1.75. 2.05, 2.41, 2.50, 2.90, 3.41, 3.88, . 5.53; dieldrlm 1.00; p,p*-DDDi 1.27; p,p'-DDTj 1.68 QF-1 1.10, 1.33. 1.40. 1.65, 1.72. 2.14, 2.59, 3.23, 3.88, . 4.84; dioldrini 1.49; p.p'-DDDj 1.75; p,p'-DDTi 1.91 Tho coluano employed were 102 DC-200 and 32 QF~1, both on , Chronosorb U, 80-100 mcoh and veto maintained at a temperature ' of 195*. The retention times of tho three principal peaks are . . underlined. The first of these, with a retention.time of 1.25 on DC-200 columns emorgoo with p,p'-DDD. Another major FC8 peak cnergco ollghtly later than p,p'-DDT on DC-200 columns. On QF-1 columns, tho compound with retention time 1.33 emerges with o,p'-DDT and tho compound with retention time 1.75 lnccrfcroo with the determination of p,p'-DDD. On this column there is no \ Interference with p,p'-D0T. Dleldrln appears as n trailing .. shoulder on tha peak with xotentlon tlmo 1.40. ' As a result of this interference, it appears that many .. of tho values of p,p'-DDD, p,p'-DDT and o,p'.-DDT reported in .recent literature are erroneous. Tito DDD voluuo originally . reported in Pacific sea birds in a paper from this laboratory . . DSW 033701 STLCOPCB4017663 ^UOUImC'/ *V* ChO * ' , , i |V> A !' rl) tt %* % i .. t . k' .* . . ' * t. (13) were coo high because'of PCB interference end have subsequently been corrected (1).' p,p*-DDE is phe most abundant ri'.\ 4,1 I* n* i .* * * .* * r.' - of eh* DDT compounds intho environment *nd`thare la no aigniflcenC PCB 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 bo greatly changed after correction for this interference. o,p*-DDT, which has been shown to be on activa estrogen (14,15) is present in very low amounts in Che global environment, although o,p'-DDE is comparatively more abundant (1,3). When both DDT and PCB peaks are present in chromatograms, the clue to the relative amount of PCB comes 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 . 1 peaks usually have approximately the same height on chromatograms obtained with sn electron capture detector. If the peak with retontlon time 1.48 on DC-200 columns is approximately ao high aa the "ODD" and "DDT" peaks, no or very small amounts of />?? * . e p,p'-DDD and p,p'-DDT oro present. When one or both of theso peaks is relatively higher than tha 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. Tho soponlfi- . .\ * cation procedure la rapid, and olnce only -a relative change In peak heights is determined, it need not be quantitative. * ' \. . . . .'. '. ' -\------................................................ . ; An \' w ' i nr intern OSW 033702 STLCOPCB4017664 aliquot of the extract Is refluxed for approximately 5 .minutes /I in ethyl alcohol with 5Z KOll, to which hexana and conccntratod aqueous NaCl oolutlon aro added In turn. Chromatography of I the hcxnnalnycr shows that tha p.p'-DDD and p.p'-DDT peaks ' have bean displaced, but the PCB poaks are unchanged. Tito contribution of PCB to the peaks of the original chromatograms can thereby bo estimated, provided the datcctor response is linear. ^j'.jPCB hno been, ohown to be a powerful inducer of steroid ^hydroxylases in birds (1) end, with p,p'-DDE,' may therefore be ` '.partially responsible for the aberrant calcium phyoiology ,.observed in apcclca of raptorial and fish-eating birds which ^accumulate chlorinate^ hydrocarbons,.(16,17)... It. is probable ' .. .that the PCB la human food supplies would Induce the .synthesis .of comparable enzymes in man............... . - v . Since it la highly^unlikely that primary standards of ^individual PCB compounds will be available in the,foreseeable ' future, the^quant ificacion procedures, like those for toxaphene (10), aloo. a mixture ofpolychlorinated compounds, must for the t...perecsent .bo a ppro-x..i.m...a...t.e.., ...T..h..a....q...u. antific'ation procedure we have . , ^wed (1,2) is based primarily, upon the.determination with a microcouloactrlc dcetector of. the total chlorino content of tho ^PCB compounds emerging after p,p'-DDE. TJho. chromatograms of . . i Ofliivs.rh. >\va.vni dWbluir*dweix..t*ra.c..t.s.....m...o..s..t,*closely rcssembla those' pr roduced .- thpo ponmorciol^aroclov, jrhich, ftas an average, qhlorlne content' ok the "DDD" nnd"DDV' peaks,, no or very'small .u.-.oiint*;. p.p'-DDD and p,o'-DDT. u^s 'present. Kvji<n r.e or both .*..* "' peaks i3 relatively hip/..e\ than '1$ .di'.i pea.< with ;-.v.` . r 1.43, the amount of p,p*-D.V) p.p'-DV*. or..'. re .............. the f'n.tr.;;ei ir. peak he <;.*. 2 /..'. . rr2: .v.. < || I l I|^<7U yp.<', in wm ^ i w, i ^ . i; in m apii . '; : '.To v . DSW 033703 STLCOPCB4017665 H .<.1. (<>.') .. , I 11 1.J *,'l fl# I* Ta'i. , il' l.t'l l *.'* I ,.bf 5AZ.`" PC3 concentrations In fioh and blrdo vero therefore ilorlvcJ from the chlorine valuea by assuming that the chlorlno ,`content wao also 54X.1 ' Thla procedure would occasionally include .. other chlorinated pollutanto with the PCB. It any, however, bo taoro meaningful to measure total organic chlorine (18) rof non-biocidc origin in tiasuca of wildlife than porta per ' Billion concentrations of Individual compounds. Title might be a bettor index of physiological effects such as cnxymo induction. , Since we do not yet hove In our laboratory the permanent . use of a nlcrocoulomotric detector, wo have had to depend upon . electron-capture for routine quantification of PCB. Each PCB : compound wot initially assumed to produce tho same peak holght, with the electron capture detector, as an equivalent amount of . 0 - p,p'-DDE. Amounts of sample injected into the gas chromatograph 'i , wars adjusted in order to fall within the linear response . range of tho electron capturo dutcctor. Tho amount of total PCD conoldcrcd as p,p'-DDE was then multiplied by a factor . t v which would yield a valuo cquqj. to that obtained with a micro- . coulomctric detoctor. This procedure wss periodically checked by electron capture determination of standard PCB mixtures. Although this method yields approximate results, it does . \ permit . accuratve \ determin.atio'ns of . relative conc' entrations. It has shown.that some species of birds, notably certain raptorial . ' 199 . ' species, of PCB (. and conci also sho-conccntr/ The and 1.33 chromato> Bay and 1 is prescr tho Culf aerial fc. Californi tested th selective in the at evaporate h dishes co / rthcn irra gcrmicida of peaks essential height of was irrad Mwu.m'm; '! pvu *.pyjsms.ti'awiwi".1 'i; r, ' . /* . * jw"**1 DSW 033704 ISMW STLCOPCB4017666 epecica, ohcarwatcra and petrels, accumulate high concentrations of PCS (1). Terminal carnivores of food chains nay accumulate and concentrate the PCB present In prey species (1). It has also shown that regional fallout patterns exist (1), with highest ,u! ''*................ * ... . , concentrations in Industrial regions. . . * * ... 1 . The PCB compound with retention tirso 1,25 on DC-200 columns and 1.33 on QP-^l columno 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 proacnc, or is pre.s..e...n..t....i.n... re latively lo'w araounco, ' in cxlrocto of birds from ' the Culf of California. PCB in this area must derive from i*% ..................; . , ' s* ..e...r.i..a..l....f..a..llout, sinco thors are few Industrial areas in Baja California or along the coast of western Mexico. We therefore . . .i.. ............................. . ` tested the possibility that this compound might have been w *. .* ...*%*................... ' -\ selectively degraded by ultraviolet light during its transport in the atmosphere. Standard PCB eolutiono in hexane wore ' , evaporated.to dryness in Petri dishes, or were applied to Petri dlahoo containing either water of 3X NaCl solution.. They vero then irradiated at a distance of. A inches with a 15-watt .... 111:. .. ,, Jl. ............. - * * . .. * .. germicidal lamp emitting at 2537. A. . In each case the pattern * ** i of. peaks on the chromatograms of the Irradiated PCB was'' 4 'v*l *1- *' ............................................ r . 1 '* * I essentially unchanged, except for a relative reduction /in the , '' ' height o this. peak.. The ..effect, was. more pronounced when FOB C 0*' a *".** a a' a t ilOV -!.*. I ,.. > *. * ' ,' * ' j ' 1 * ` i ' ' ' #*'./, .; ` ; yaa Irradiated on .the, aqueous surfaces,.\V. .'-J ................. i)V wvk . " ................. ** - - - . v :........ . . . ;: . . . . . Although thiAyoV.iod j'loid.-:. / permit jocvk'ACc doto.Xi.'.Atiooi of relative v/oi . . I : ' ' ' ;i ; ; . .I hos frtjf.M tV.-.t Jono jrpcc'ou or' :..v: ably c..j;`t..n: . - ' . . * ' * \ '`. . \. ' ' ' ' r'H'J iif ii .,f . r ,* * \ . 't ' . osw 033705 ' . , / . STLCOPCB4017667 Acknowledgements Vo thank tho Monsanto Chemical Company for supplying aamplos of tho Aroclora. Tho National Sclooco Foundation (GD6362) provldod financial oupport. . Roforoncos 1. R. w. RISERROUGH, D. B. PEAKALL, S. C. HERMAN, H. N. KXRVEN and P. RIECHE, Naturo 220, 1098 (1968). . '' \ 2. R. W. RICEDROUGH, Chemical Fallout, Flrot Rochoster Conforonco on Toxicity, In tho proas. \ 3. n. W. RISEERGJGH, D. B. KSNZEL, D. J. MARTIN and H. S. OLCOTT, In preparation. ii. Tho Poctlcldo Rovlow, United States Dopartmont of Agriculture .. (1966). . .... . $. E. S. NIELSON, Ann. Rov. Plant Physiol. 11, 3Ul (i960). , 6. F. ACREE, Jr.. M. J3EROZA, and H. C. BOWMAN, Agrlc. Food Chem. 11, 270 (1963). ' 7. C. WIDMARK, J. Ass'. Off. Anal. Chom. $0, 1069 (1967). ' ` 8. D. C. HOLfES, J. H. SIFMOHS, and J.O'C. TATTON, Nature 216, ' 227 (1967). . . '' ' . . . 9. A. V. HOLDEN and.K. KARSDEN, Naturo 216, 127U (1967). 10. T. E. ARCHER and D. 0. CROSBY, Pull. Environ. Cont. Toxicol. 1, 70 (1966). . :. 11. F. ERRO, A. DEVOUR, and H. BECKMAN, Bull. Environ. Cont. Toxicol. 2, 372 (1967). , . ; . ' 12. M. S. SCHECTER and H. L. HALLER, J. Am. Chom, Soo. 66, 2129 (19W). 13. R. E. RISEBROUGH. D. B. MENZEL, D. J. MARTIN, and H. S. OLCOTT Naturo 216, 509 (1967). .. ' Hi. R. M. WELCH, W. LEVIN, and A, H. CONNEY, Toxlool.' Appl. . Pharmacol., In tho press. 15. j. BITMAN, H. C, CECIL, S. J. HARRIS, and G. ?. FRIES, Solanco 162, 371 (1968). 16. D. A. RATCLIFFE, Naturo 215, 208 (1967). 17. J. J. HICKEY and D. W. ANDERSON, Science 162, 271 (1968). 18. F. A. CUNTKER and J. H, BARKLEY) Bull. Environ. Cont, Toxicol, 1, 39 (1966). \ . '' . \ 301 . I \ /T