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i i 2 t * ;. -0. i l ?i %i II (. / *f 71 ^ - - --- - .iwilh>Mi.. i-^itansa i i - Pejtldde residue nIy s In the presence of Polychloroblphe lyls (PCB's) By ' L. M. Betsouk* 1 i Content: i Introduction ....... tt II. Presence of PCB's In the environment.................................................31 \ ! Ul. PCD Interference with pesticide resldi e isilytli.............................. 3ft IV. Uses end properties of PCB's . . ................................................ 34 V. Possible modes of entry of PCBi Jnt< the ecosystem ..... 34 . VI. Toxicity of PCB's......................................................................................... 36 VII. Analysis of pesticide residues in the pr -seoce of PCB's.............................37 VIII. CoonriMtiop of pesticide Identity . 38 IX.Identification of PCB's In samples ....................................................... 40 X Estimation of PCB's.......................................... 44 I XI. Separation of PCB's from organo^ Hoephorus compounds by the Florisfi technique............................... 45 XII. Residue* sf orgenoehlorine pesticide! tod PCB's In Canadian wildlife 45 Summary................................................. 51 R4*um4....................................................... S3 i; Zusammenfassung..................................................................................................... 54 Referenee* . 54 I. Introduction Although there has been to much discussion on the pros and cons of pesticide use, it would be unthinkable to start this review without ft tew comments on the controversy. The recent banning of DDT by tome countries and the widespread and increasing animosity to* wards Ae use of the organochlorine pesticides in genera) are due to number of factors including: a) Hie large annual additions of the pesticides to the environ ment coupled with their apparent persistence. b) Their known acute toxicities. c) Their widespread distribution in biological materials and their reported buildup tn the food chains * Ontario Rosearch Foundation. Sheridan Park, Ontario, Canada. J7 I I IT" HONS 06651? ~T] ,,./8 w----- 18 UM. lUn olm d) Their more recent implication In the derangement of oalchtm metabolism (Peakau. 1967, Si no* 1967, Welch if at 1969) possibly causing the decrease in eggshell thickness and the population decline of certain raptorial species of birds (lUTCLtm 1967, Hickey and Akdehson 1968, Sticxel 1969). a) Their adverse effects on fish rep eduction (Bunnies it el. 1964) and high residue levels found J i Coho salmon in Lake Michlcan in 19G9. f) Certain "fears* involving ebroni* toridtie* and sublethal doses with possible deleterious effects on man and wildlife and the ultimate survival of man. The above factors are very compclli ig and certainly warrant man's concern about the threat to his environment and his very existence. Nevertheless, one cannot deny that the c pesticides have been impor tant in ensuring the supply of man's k oidly increasing food require ment and in the protection of his herhh, as well as the saving of millions of lives. Surely, this Is a case w icre the benefit-risks equation is well as the alternatives to the org mochlorine pesticides should >e carefully studied and evaluated to a *okl the possibility of getting nto a worse predicament. It was quite ironic when recently a large city publicised that, tccause of an impending ban on DDT, the garbage collector would ]>ick up separately all home garden sprays and other materials which "Or.tsir-cd ewa traces of DDT. The irntrrlal that had been used store than generously And cherished for i Imost 30 years had suddenly l e some monstrous even In trace miantitir?. Wasn't there much greater danger posed to the collectors, children, and animals by such collec tion than if the home supplies had been flowed to run out gradually . with no chance Of replenishment? In fact, the announced bans on DDT so far are generally not total, and hence serve the same purpose as merely tightening the control on the ,utet of such pesticides. In other words, when it is necessary, and until suitable and satisfactory replacements are avail able, limited Mid Judicious use will be made of these pesticides. With regard to the use of substitutes, caution is essential to avoid ?rfmature use of any compounds. It is now known that potentiation or antagonism) of toxicity Is possible with certain combinations of pesticides and with combinations of pesticide and drugs or environ mental chemicals (Dubois 1968). A compound might be fairly safe by itself yet hanhfut in the presence of others. It Is a fact that even il suitable substitutes were found and the mo of the organochlorine pesticides was banned completely, a need would still exist for at least a number of years to have reliable analyti cal methods for die determination of their residues. The present re view discusses the status of organochlorine pesticide residue analysis HONS 066516 I derangement of < NT, Wnat * oL 19fi9) pheO thickness and the oital spades of btrdf a* ISM, Sms. I9fi9). lag (Bukmcs <( el ISM) salmon In Lake Mlohl- itiet and sublethal doses wn and wildlife and the t and Ua vaav Mi ittddea have been I toomaafag food require- M writ u die laving of lha beoefft-riiks aquation hlorfae portddm abanld dw possibility of getting gl Oily publicised that, garbage eoDactot would aw other materiab which rial that had been uaad aat 30 yaars had suddenly Vaat dura much greater d anhnab by uch ooOeciwod to run out gradually ao far are generally not ai merely tightening the Other words, when It la y roplacements an avail- ada of thoia pestfcfdea. nitton la uaaitlal to avoid r known that potapHation h certain aomhlnationa of Ida and drum or eoviroo- anad might be fairly tale wan found and I completely,idli in to have rjhaWa wMwt> . putkm raridue tntlyiii . MydiWtitqilwyh to with particular reference to lertoua Inaocuradea oauaad by the passanca of polychlorobipheoyla (PCB'a) In tome samples. At would bo expected. In order to cope with the numaaoua new peeUddt* that have bean put Into uae, pesticide raaldua methodology has changed drastically over the yean. Zwso (1983) pointed out that until about 1940, the Ufa of an analytical chemist working In the peatidde Sold wu a relatively serene one In which the chemist had to be familiar only with enidyticel methoda for arsenic, lead, Suorido, pyrothriru, ratenone, and a few others. Juat about tan yean back, DDT and other organochlorine compounds were eetiinated by eolorimetrle methoda (Somcim at al 1945) which obviouily did not give precise Informatioo on the amount! end ldentitieo of the indmdual compounds present Today's residue methodology Is much mon complex and In ardor to cope with die new pesticides being introduced, the techniques an constantly being modified and Improved. The wide variatfooi In toxicity and penlxtenco of petdeldec, new or old, dictate refinemanta to methoda In order to differentiate and determine Individual residues. Bearing in mind the complexities mentioned above and the feet that the spray history of moat sampler {excluding controlled spraying experiments) It unknown to the analyst, the eveuebUlty of chromato graphic techniques which afford separation of number of compounds fa a mixture lias made the multi-residua screening approach' (V. S. Food mtd Drug AdminhtratUm 1969 revision) the method of choice. The gas liquid chromatographic (CLC) separation with electron capture (EC) detection la presently the most popular analytical com bination for organochlorine pesticide residues. Multi-residue analysis Involving CLC generally Involves five orinripal steps: a) Sampling to procure a representative aliquot of the sample. b) Extraction of the residue with a solvent (usually orXante linos most of the orgenochlorines (OCs) and many of the organo, phosphates (Ors) are fat soluble). e) Cleanup of the extract to remove Interfering materials such as tots, warns, pigments, etc. d) Analysis of the deened-up extract, generally by CLC-EC technique. o) CoalnMtkxi of the pesticide Identity. Certainly there are a great number of variations la carrying out these itepS, depending on the nature of the pertldde and its substrate, ' Bdker's eotei Sac Icsua, I. Ai Davalepewet of the Toed tad Drag AdmiaktrWtca'i method of analyse hr mnlUpt# mMim of osgaaocUorias pad- sSdss k foods and fssda kssldus lUvWws, this vehew. HONS 066519 *0 L. M. IUtmolm the limit of detectability required, tod the laboratory ttafi and equipmen*, available. It la beyond the aerpe of thil review to dlscusa any of theae metboda in detail Furtheim.ire, Ctomm (1962) and Samos. and Hodcd (1907) have reviewed hit topic adequately. Accordingly, thir review will deal mainly with problem! involving PCB'a and re lated compounds. No attempt will te made to review gu chromatog raphy for pesticide residue analysis. Burnt! (1965) has discussed tome practical aspects of this topic, while 7 iiompson el(. (1969 and 1969 a) Lave evaluated different gas chroma'Ographic columns for chlorinated pesticides. Before discussing the PCB's and their interference with pesticide residue analysis, some1 hey referee sea In the English language on residue methodology are appropriat >. Theae are listed alphabetically u follows: 1. Advances in pest control reseat ch, vols. 1-6. New York: Inter science (1957 < tea.) 2. A guide to the analysis of pesticides by gas chomatography, 2nd ed. S. T. Preston, Jr. Evanston, 11..- Polyscience (1966). 3. Analysis of insecticides and sea. icicles, Gunther and Blinn. New York-London: Intersclcncc (195!). 4. Analytical methods for pesticides, plant growth regulators and food additives, vols. I-V, Zwcig. Nev York: Academic Press (1963). 5. Guide to the analysis of pcsticid > residues, vols. 1 and 2. Burch field and Joltnsoo. V. S. Public Health Service, Office of Pesticides, Washing:;* D. C. (1535). 6. Manual of methods for the determination of residues of Shell pesticides. Shell Chemical Compciu, Agricultural Chemicals Divi sion, New York (1969). 7. Official methods of analysis. Association of Official Analytical Chemists, 10th cd. Washington, D. C. (1965). 8. Peatictdo analytical manual, vols. 1 and II, U. S. Department of Health, Education, and Welfare, Food and Drug Administration, Washington, D. C. (1966). 9. Pesticide residue analysis handbook, Boneili. Wilkins Instrument and Research, Inc., Walnut Creek, Celif. (1965). 10. Residue Reviews, vols. 1-34. Cunther. New York: Sprlnger-Veslag (1962 << seq.) It should be emphasized that these references were all written prior to the knowledge that there was a possibility of PCB Interference with 'Editor's note: Them an bow more than 05 books eonrented with pesticide nsidoes Including 60 that discuss annlyticai methodology in soma respects; eight eountries are nprnscntrd by tha authors. Cormraa, P. A.i Pesticide newues to rha total eavlronment-lleliable detection and detenotaition, mitigation, and Weistattoo oootral aad sumtUanco programs. Pun and Applied Cheat. II, $55 (1170). nans 066520 m PoIychioroblpbaDyli 91 certain pesticide residue analyses. To general, the methods described In these references can be modified ro cope with PCB Interference, IL Presence of PCB s in the environment Following the development of the EC detectors and their use with CLC, most residue chemists have observed unidentified peaks 1 on their gas chromatograms Generally, there was no specific pattern to these peaks, but analysts concerned with pesticide residues in wild* life, especially in Europe (Roou.e 1965, Haaiuson 1966, Holden and Mamdb* 1966), sometimes observed a particular scries (with as many as ten or more peaks) of the uni lcntified peaks in their sample ex tracts- The mateiials giving rise to tiiese unidentified responses occurred most frequently and in the largest proportions In extracts frojn aquatic raptorial species of k rds and fishes. The general supposi tion was that these responses were from condensation products of the metabolites of the organochlorne pesticides, since they were gen* erally observed when large ainotu ts of pesticides were also present. Roiiuhy (1965) reported them to e organochlorine compounds with k fairly high chlorine contents, vhic \ no doubt accounts for their high sensitivities to the EC detector. Iowcver, Jensen (1966) was the first to state that these unidentified peaks corresponded to PCB com* pounds, baaed on the analysis of a number of pike samples, an eagle, s and human hair. Initially, the CLC and thin-layer chromatographic (TLO) pattern* and chemical inertness were used to identify the unidentified peaks as PCB's (Je\sk> and Widmark 1967). Other workers in Crest Britain (Holmes et al. 1967, Holden and Mammen 1967) and the Netherlands (Koeman ct al. 1967) also mode early reports on the presence of PCB's In their fish and wildlife samples. At the Organisation for Economic Co-operation and Development (OECD) Conference on "The Unintended Occurrence qf Pesticides in the Environment,* held in Scotland in September, 1967, the Euro* peon chemists in particular showed much concern regarding the pres . ence of PCB's In their wildlife samples and the possibility of inter* . ference with their organochlorine pesticide residue analysis. Up to * > that time, tl^ere had been no report on the occurrence of these mate* rials in North American samples. However, in a December, 1967, reporti on Organochlorine pesticides in seals and porpoises, Holden and Mampen included a comparison of the amounts of PCB-type materials fopnd in Scottish and Canadian samples. They reported that the levels of these non-hvdrolysable materials were generally much lowetr In the Canadian specimens. Since that time Risemovch at al. (1968) have reported that the PCB's are widely dispersed in the global efcosyatem, while Reynolds (1968, 1969, and 1969 a) has * found them In some Canadian wildlife samples. i HONS 066521 U M. Ritnolm Positive confirmation of the PCB` by gas chromatography-mass spectrometry (GC-MS) has no\ been reported in Sweden (Wiomiut 1967), the Netherlands (K0EM4N tt al 1969). and` in th* e United States {Rejchel 1969). lit PCB Interference with pesticide residue analysis Uni polychlorobiphenyls (X indicates the possible chlorine post* Uoni) and related compounds (polychlorinated tripbenyis. naph \-- t(/ *h--5 mx thalenes, trrpcnes, etc.) have &u> merous important industrial uses but are not used as pesticides. Because if their similarities in structure and (iroperties to the DDT pesticide group, tho PCB's--if present In i substrata--are carried through the usual pesticide extraction and s reening procedures, and since thev possess electron absorbing prep Mtios, will interfere with CLC-LC analysis of orcanoclilorine as wi ll as a number of the organophos* phorus peatldocs (Reynolds 196! b). The reported GLC patterns indicating PCB interferences have shown marked resemblances wil.i 'Arodor' 1254 and 1-60 (Icnsln* and Wipmmuc 1967, Holmes et of. 1967. Rlysolos 196$ ana 1969, and Koeman el at 1969). This type of interference with pesticide residue analysis is demonstrated u iili standard mixture ot orgunochlorine pesticides and a commi-rcinl PCB mixture (Arorlor 1254) in the chromatograms of Figure L The results confirm (hit the pres* ence of FCB'f m the sample extracts will cause interference with pesticide residue analysis under these or similar operating conditions. It should be emphasized that throughout this review when the term ^PCB" Is used, this Includes the polychJorobiphcrrls and also the other closely related compourds mentioned above. However, the polychlorobiphenyls are the most important from the point of view of interference with pesticide residue analysis. Under the GLC operat ing conditions, the chlorinated polyphenyls and other related com pounds with molecular weights greater than the chlorinated biphenyls will generally not be separated and obaerved on the gas chromato grams. Modification of trie operating parameters such as higher col umn temperatures or changes in the liquid stationary phases are usu ally necessary to detect the presence of such compounds. It must be borne in mind that when high molecular weight com pounds are present, their peaks can appear on subsequent chromato grams and cause "secondary interference." This occuner.ce can be recognised easily by the early emergence of unusually broad interfer ing peaks on tho chromatograms. "Secondary interference" is a term used by the author to imply that the interfering peak does not have MGNS 066522 U i ' 1 Folychlc -oblplKoyb 33 the lime retention time u that of the compound of interest under the tame operating conditions. "he interfering peak often originates bom a previous injection and ca rnot be simply repeated. Of course, if the elution time is very long, the peak might be broad enough I to escape detection and in such a case would not interfere. In contrast m > i+ \z iS Pig. 1. FCB Interference wfth ovganoct forint peitickfo residue analysis on four percent SE-90/sk percent QF-1 on 00/80 mesh Ckromosorb W, V x * MwlHotlt column. Chmnilo|rjm A: Unditd mixttm of orienochloiine ptMicldai peck numbers: 1 0 08 n|. ol lindane, 8 0.10 ng. of nentachfor, 3 0.10 ng. of aldrtn, 4 0.14 ng. of beptacMor epoxide, g 0.80 m. of DDE. 6 020 ng. of dleldrfo, 7 - 0.90 ng. of ODD. . and 8 0JO ng. of ODT. Chromatogram 8.' 5 ng. of Arodor 1854 (tbo . 14 major peaks ara numbered I to aIV). Chromatogram C: combina tion or above oifanoehlorlno standard paattcldo mixture and Arodor 1884. Injector 8SCTC., column 400*C., detector bw 150*C, N. at 80 ' 80 eo/mlouto . to this type of Interference, materials which have identical retention 1 times will dause "primary interference" which is easily repeated by reinjection of the extract. With this form of interference a single, ' shaip peak Is obtained, and in the absence of confirmatory and other precautionary measures, misinterpretation can occur easily. PCB-type interference falls mainly in this group. I *QhS 0665*3 .'4 L. M. Ritn>ih IV. Uses and properJei on PCB'i In order to understand how it U p >ssib)e (or PCB's to be preteat in wildlife end cause Interference with pesticide residue analysis, it If necessary to give a brief description of some of their properties aod uses. The PCB's are produced and ma keted under a number of com* merdal trade names, e.g., `Arcelor/ *Cophcn/ end `Phcnochlor.* Koxman at al. (1909) reported marked resemblance between Aroclor 1260, Clophen A0O, ana Phenochlor C P0. The `Arodor' plasticizers, a series of chlorinated biphenyls and chk.lnated polypheny!*, are typical nf these compounds. As stated by he manufacturers (Monsanto Co. 1907), the 'Aroclor' compounds are among the most versatile chemically produced materials available. They vary from mobile, oily liquids to write crystals and hard transparent resins. They Are nonoxi dizing, inert, permanently tliermopljtic, of low volatility, non corrosive to metals, insoluble in water, and resistant to alkalies, adds, and corrosive chemicals. The viscous, nore highly chlorinated liquid and rosin members do not support combustion and they impart fire retardaoce to other materials. The c ystallme `Aroclor compounds are relatively insoluble but the liquic and resinous compounds are soluble in most of the common organic .olvents, thinm-rs, and oils. These compounds are used in protective coatings, as placticizers and extenders, as sealers in watcr-pi aofing compounds and putty, in asphaltic materials, printing inks, waxes, and synthetic adhesives. Liquid PCB*s are used as dJdcctrics, ns hydraulic fluids, in thermo stats, in cutting oils, as extreme pressure lubricants, u grinding fluids, and as heat transfer media. Solid PCB's are used to impregnate carbon resistors and as sealers In impregnating agents for electrical apparatus. Tho properties of many products can be varied and improved by tibe use of these compounds either as primary or secondary plasticizers. Further details concerning the different `Aroclor* series, the grada tion of their properties, and the numerous possible applications can be found In the rqanuficturer's technical bulletin (Monsanto Co. 1907). In the system of designation for the 'ArocVon,' the first two digit* indicate the type of material, while the last two digits give the approximate wergnt percentage of chlorine in the product. For example, 'Aroclor* 1254 indicates a chlorinated biphenyl with approxi mately 54 percent chlorine, while `Aroclor' 5400 indicates a chlorinated triphenyl containing about 00 percent chlorine. V. Possible modes of entry of PCB's Into the ecosystem The means by which PCB's enter the ecosystem to contaminate fish and wildlife are still not clearly understood. There are however, . r H0*S 0665^4 Polychl iroblphenyls 38 four most likely pathways by wh ch PCfi'i enter fish and wildlife: ) Because of their lneti ess and versatility and consequent numerous applications, i: is quite conceivable that PCBs could be flushed as was es into rivers, lakes, etc. to pollute fish and other wildlife. b) There is a possibility ths: some contamination could proceed . via the atmosphere wher wastes containing these compounds are burnt o) Although it is unlikely, .ome type of Ullman reaction with condensation of aromatic halides with the aid of metallic agents such ss copper to give, rise to the formation of biaryls, cannot be completely nil<4 out. " d) Lastly, but not nccessarl y the least likely, there is the possi bility that some companies are using PCBs in some insecticide formulations to Increase t ie kill-life. The idea of increasing the n sidual persistence of insecticides his been investigated for some tin e and a brief r&umd on the more important background work is in order. The early investigations by Lindquist ct el. (1945) and Block (1948) indicated that the rcshlual effectiveness of DDT was pro longed when the DDT was ap died in resins and paints, van Tiel (1952) continued along these lines when he increased both the effec tive ceded and the toxicity of a DDT deposit ca a glass surface by the addition of a small amount of coutnorone resin to a DDT Solution in kerosene. Subscque itly, emphasis was placed on work involving the more volatile insecticides like lindane--a very effective pesticide but one which soon loses Its activity because of volatilization. Sullivan and Hornstein (1953) in their experiments with the American cockroach, Perlploneta armrlcana, found that lindane resi due remained toxic longer when it was compounded with `Arodor' 5480. They suggested that the chlorinated polyphenyl prevents crystal lization ot the lindane and lowers the vapor pressure of the lindane in the mixture, thereby preventing unsightly residues on surfaces as well as extending the effective kill-life of the insecticide. Tsao ft of. (1953) obtained similar results against house flies, Mutce domeslipe Li Continuing this work, IIornstbin and Sullivan (1953) con firmed these findings and described a general method for prolonging Ae residual effectiveness of volatile Insecticides. The method consisted of preparing concentrated solutions of the pesticides in the film-formlog polychlorinated polyphenyls. A Monsanto Co. Technical Bul letin (1965) states that because "'Aroclors' in formulations *txap* and hold more volatile ingredients, they make volatile insecticides and repellents last longer* in residual activity* Attempts to determine whether this idea had been put into practice have so far been unsuccessful. However, If the PCBs are being used MOMS 0665*5 >--- M L. M. lUrvoLM I In pcstidde formulation* then tVi coupled with the numerous other uses suggested in later Monte 40 Co. Bulletin (1967), might cer tainly. explain their presence in wildlife tissues and other samples. There h ail obvious need for clarification of the sources of PCS'* in'fish -ana wildlife* and this oould probably be accomplished using tracer techniques. * VI. Toxic ty of PCBt Tito PCB's were studied at coily as 1SSI (Schmidt and Schultz) and were in wide use bv 1930 (Pcvnixc 195.0. Later lovu and Aloe* (1936) reported tnat the compounds were toxic. Gmxniem ' et oL (1939) reported that PCD* ana polychlorinated naphthalenes usedltho deaths of three workers. Men employed in the production ad usNhf PCB's developed acnc* ype skin crupuoni (Schwartz and Barlow 1942, Schwartz and Peci 1913* Schwartz 1943). Xtiuxn (19U) reported that 5CB`s caused pathological changes In laboratory animals. Brown (19 7) warned about the toxicity dan* gers of the `Aroclors' when there was a su^'-on that one of the Arodors* be used as the melting point, bath jit uid in preference to th# customary sulfuric add. Later McLait.juliv rt at. (1063) found . tHftt Aroclor 1242* besides being toxic, produced teratogenic effects Ih chick embryos at lower dosage levels. l\i*T*r ouch et ol. (1968) reiteratpd that PCB's along with othr uMutuia.rj biocides, such as DDT, .could account for a large part of the a: rrrntion in calcium metabolism that has been observed in manv species since tho second world war. With high dosage of PCB's* byaropericards have been (Observed in quails (Koeman et ol. 19691. It ihas been stated (Monsanto Co. 1967' thit at ordinary ternIporatures the Aroclor* chlorinated poKphenyU d : not present Indus* * triiltoxicologkal problems. However* caution was idvised in handling thesf materials. The hazard of potential toxic exposure varies with ft|e volatility of the compounds: the lower chlorinated* more volatile ones present more of a potential problem from the standpoint of { both Inhalation and skin contact. At elevated temperatures, the use of PCB's requires veiy effective and efficient exhaust ventilation systems. Irhas abdbeen stated by the manufacturers that 'tests on animals I Indicate that the maximum safe concentration of v.-por is in the range of fitom 0.5 to 1.0 milligram of the lower chlorinated Aroclor' plastielzeiis per cu|ric meter of air. The threshold limit* f maximum allow- 4>le^ concentration for an 8-hour working day) set by the American ConipfftiKe of Government Hygienists ore 1.0 m:cram of the lower chlorinated 'Aroclor compounds per cubic meter c: air and 0.5 mill!* " the more highly chlorinated compound*, such as Aroclor' cubic meter of air.* * '.it HONS 066526 <r < ir -J Jtc .4t .J fit* t the v to '4ftd Hots .*) h as Hum vood . been tern* Indus-idtiiig > with olatile sat of tl* use illation uiimals c rangd t* plastit allowAmerican ** lower MmiUlAtodot' ! i I Polydilorobi >bcsyla 97 However, as Knesnovcit ef al (1968) pointed out, only relatively imall amounts of chlorinated hydroc irbons are required to cause en zyme-induced breakdown of steroid i, thus making irrelevant much at the p.p.m. approach to polluted ecology based on toxicity data alone. .t There Is no doubt that further .esearch in this area is urgently needed Vll Analysis of pesticide residi es in the presence of PCBs PCB's are of interest to analysts for three main reasons: * a) The compounds arc toxic. b) They are widely distributed ind their uses are increasing. c) They interfere with CLC determinations of organochlorlne and organoplrosphorus pvstici' (e residues. The first two reasons havo been dealt with earlier and therefore need no further elaboration. Accon ingly most of the remainder of this review will be concerned with he analysis of pesticide residues in the presence of PCBs. Also tin present methods of estimating ' the amounts of PCB's will be discusst d. As mentioned earlier, it was rmphaftfoed fit the 1967 OECD confer ence that K'B's were in fact bcin; detected in fish and wildlife, especially in Europe. The conference among other things served to alert other chemists and scientists to the problems of PCB's and their interference with pesticide residue snalysis. Since the discovery that PCB's arc contaminating fish and wildlife and that they Interfere with organochlorinc pesticide analysis, very little has been published on how to cope with these interferences and how to differentiate tho PCB's front pesticides. Based on preliminary work, Jensen and Widmauk (1967) suggested a nitration technique based on the Inert ness of the PCB's compared to the relatively easier nitration of the t pesticides. Kusanovctt et al. (1998) reported the use of this method, < but Reynolds (1969 b) pointed out tnat nitration is not a suitable approach since*some pesticides (lindane, toxapenc, Strobane, etc.) apparently will not nitrate, while some of the PCB's appeared to , be nitrated under the suggested conditions. In addition, there is the ' problem of further complication from the interference of the nitroderivatives formed, especially when the DDT-group pesticides are present in large amounts. To avoid the above complications and at the same time facilitate tine chromatographic interpretations and afford a means of estimating the PCB's and pesticides independently, Reynolds (1999 b) Intro duced a Florisll separation scheme. Koemax et al (I960) have also reported a similar Florisll column technique to separate the PCBs ^ from tne pesticides. HONS 066327 t .> * T M 38 L. M Riikolm In Reynolds' method, the pa (tally clcancd-up sample extract con taining the pesticides and PCR*. Is placed on a Florisil column and the PCB's arc eluted first with nhexane, followed by elution of the pesticides with an ethcr-hexam mixture. Originally, this technique was intended to serve as the fit al cleanup strp, as well as effecting separation of the PCB's from the pesticides. As will be explained, the method has since been sligi tly modified (miniaturized) to serve mainly as a separation technique Of the many pesticides Investigated, only ncpUchlor, aldrin, and the )DT metabolite ;>.p'-DDE are eluted with the PCB's. With the except: on of p.p'-DDE, these do not present difficult problems since chemical reactions (epoxidation, hydrobromi- nation, etc.) can be used to diflrrentiate heptachlor and aldrin from < the Pt^B's bv this scheme. p,pf-DDE on the other hand presents a more difficult problem on account of its chemical inerness. Its greater stability resembles that of the PCB's, and the com} ound Is not amenable to any simple and Convenient method of strut hire modification to give a product which if readily detected within the usual CLC-EC operating param eters. ' Of course the ideal situation would be a convenient addition of ond mole of hydrochloric acid o the /,p'-pDE to form p.p'-DDT While the PCB's are not affected. This would afford confirmation and quantitative1 estimation of p.p'-LDE in the presence of PCB's. Cen- . erally under such circumstance* it would not be necessary to do an ' additional Florisil separation. Hu- amount of p.p'-DDE present could be estimated indirectly from the ,;/*DDT produced since, as will be discussed later, the p,p'-T>Vl (and p.p'-DDD) can be separated I from the PCB's by the use of highly polar liquid phases. * It phoujd be emphasized that although p,p'-DDD and p.p'-DDT are mostly separated by the polar phase columns, without introducing I; a Flprisil separation the earlier emerging pesticides are superimposed oq the PCB's and hence a separation is necessary to identify and to quantify these latter pesticides. After the Florisil separations of PCB's and pesticides are made, the two eluAtcs are chromatographed separately on the SE-30/QF-1 coluiph and compared with appropriate standards for quantitation. ' This It n^irjafiy followed by confirmation of the identities of the pesticide*, j; Ito the separation of the PCB's and their elimination as Interference, the non-specific nature of the EC detector i MOKS 066528 ' PolyclkJorotripbenyb 9o maket It necessary to coofirm the ^entity of the pesticides by addi tional techniques. The sequence usually followed by our laboratory is: . a) Injection of the pesticide eluate onto a more polar liquid phase column (e.g., polyester such as DECA or DECS). Typical *! ft : :*> Xt DT nd ra in iM *11! sled :>dt oCQ Md ade, lien. (the M t*tor Fig. t. Separation of PCB'i and organochlorlne pesticides ca polar phase ooiumn (five percent DECS/two percent HiPOt). Chrc-r.awgram A: standard mixture of organochlorlne pesticides with fame a.*: runts as In Figura 1; peak numbers: 1 hrptachlor end eidrin. 2 * i;.-. ;ane, 9 heplaehlor kJe, 4 " DDE, S dieldrln, a DDT, ? DDD. Chromatogram n u, of Arodor ISH (the 14 mt|or peeks ere numbered I to XIV as In chromatogram B of Figure 1, assuming tin*. tl>e order of elution is unchenged). Chromatogram C: combination of cr janochlorlne standard * . pesticide, mixture and Arodor 1954. Other DLC parameters as in Figure 1 separations of pesticides^ PCB's, and a mixture of the two groups lire shown in Figure 2. The reversal of p.p'-DDD and p.p'-DDT and their separation from most o: the PCB's Is note worthy and quite useful. One drawback is that peak No. 13 of Arodor 12o0 interferes partially with p.r'-DDT, Other use ful information can be obtained by study of the retention times y I i HONS 066529 U : 4. Hsymolds of Individual pesticide* Foe example, this Approach affords separation of the throe main B2IC isomers. Tm technique is ,1 especially useful for riling out the presence of pesticides in* ,1 dilated as possibly pre* ?nt by the SE-30/QF-1 column. How. ever, the retention tim * on two or more stationary phases cannot be regarded a: independent parameters of identity (Robinson 1907). b) DerivatizAtion and use of characteristic CLC retention times of the derivatives. As n ported earlier (Rlynolds I960 a), our laboratory has increasln $ly utilized chemical reactions for con* ( firmations of pesticide residues because the samples to be analysed are generally mall and the amount of residue is in sufficient to allow effec ive application of infrared, mass, and other spectroscopic met tods. The dcrlvatization technlqu j has been used by a number of work ers (Cwnrcn 1962. Kuhn t il 1064, Hammckce et el 196S, Saks 1967, Dumr and Wong 1967, C iadciiuk and Wa.nl*s 1969, CoainAKB and Chau 1908, Chav 1909, C iau and Cochiunk 1909, Wiencke and Bubke 1909) and is gaining i 1 popularity- Success of the tcclmique usually depends on the specif c derivative remaining lipophilic and therefore extractable with the organic solvent (usually hexane) for direct analysis bv the usual CD J-EC method. A number of chemical rcnc ions is available, but two of the more broad spcctium t\ pc* that arc quit* useful for the pesticides generally found in wildlife are: (i) Reaction with etlmnolu* potassium hydroxide (mainly dchydrochlovination) to gve produets with shorter retention times. This is effective for DDT, DDD, and their isomers, as , well as and >-DHC; the /9-isomer is not affected. 00 Reaction with hvdrogtm bromide/acetic anhvdride reagent to give bromohvdrin or bromoacetoxy derivatives with longer retention times. This reaction has been found useful for hep ' taahlor, aldrin, heptachlor cppxidc, dieldrin, and endrin. 0) Thin-layer chromatography. In spite of ccitain drawbacks with TLC for confirmation oi small amounts of pesticide residues (Reynold 1969 a) the technique is still valuable for the con* 'finriation of DDE. TLC can oc buitc useful also for tome group separation, as an ancillary cleanup technique, and for semiquantitative estimation of some pesticide residues. DC. Identification of FCB*s In samples I fata obtained by our laboratory in the course of work conducted (<, The Canadian Wildlife Service have demonstrated that PCB's am present In some but not all species of Canadian wildlife. Undoubt* MONS 066530 J i Xb llo*' . .ntlty Um* i*, our r no. to be f Ii In- fwork.I, Sani simhi ".ixcind rhnlquo iil.c And une) for the more generally -nly dehy- idontlon ininen, u !- reagent 'ilh longer ` il (or hep Midrln. Iwd with r reelduec Outlie oon'> lor eome j *. and for 'Ml. |l j , . ;j i| j : ! | j !: { , I 1 "I woducted ;1 Aat PCB'i ;fi Indoubt* I Foiychlorublphrnyls 41 edly the pretence of PCB's h t complicated the pesticide residue methodology, but this complication cannot be avoiaed If meaningful results, are.to be obtained. Since not all samples contain PCBs, it would be unwi*o to do a Klorisil separation on all extracts before having some indication whether or not the separation is necessary. Accordingly our laboratory user; the approach described below. The sample to be analysed *s extracted, cleaned-up, and assayed using the $E-30/<JF*l column. Depending on the EC results, a deci sion is made whether or not to make a Florid] separation and clieck further for the presence of PCll's. This decision is based on the as sumption that if there are no apparent pp'-DDD and p.p'-DDT peaks, then the presence of PCli's would not be expected. The DDD-DDT combination is chosen as the c Itcrlon because peaks fight and ten, two of the larger peaks in Arocl >r 1254 and 1260 have identical reten tion times with p.p'-DDD and `>,p'-DDT, respectively. Also, the PCB components giving rise to peal s eight and ten ore less likely to be metabolized than the earlier et icrging compounds. Itencc, if PCB's (1254 or 1260 pattern) ore pn'ient, peaks should show up giving apparent p,//-DDD and p.p'-.)DT values with the EC detection system. Other useful information regarding the presence of PCB* can be obtained from the initial gas chromatography. If the CI/C pattern (prior to PCB separation o:t Fi**ruu *k<>w h'gh "apparent" p.p'-DDE with little or no p.p'-DDD and p.p'-DDT present, then ail or most of the "apparent" DDE Is probably "trueJ DDE. Tills Is because peak five (the peak which interferes with p.p'-DDE) in the common PCD commcrci.il mixtures (Aroclor 1254 and 1200) is relatively small compared to the DDD- and DDT intcrfering peaks (eight and ten) as shown in Figure 1. TIus rule has been borne out by TLC confirmation of DDE In a number of such cases. As mentioned above, the two most commonly found PCB types In wildlife samples resemble Aroclor 1251 and 1260. Although tnese two Aroclors are quite similar In many respects, certain differences In their CLC patterns, as shown in Figure 3, cap be used to identifv one from the other. With a higher chlorine content, Aroclor 1260 thow*s about 17 major peaks fSE-30/QF-l column) compared to 14 with Aroclor 12-54. Besides this, one other obvious major difference Il the peak-height ratio of peaks ten and 13. This ratio has rough values of 9.3 and 1.0 for Aroclors 1254 and 1260. respectively. Additional information regarding the PCB mixture present in a tamnlo can be obtained bv comparisons of other peak-height ratios of the PCB standards and the PCB's (n the sample. However. It must be borne in mind that in practice one might be faced with complex mixtures of PCB'i rather than a single, specific mixture. In such cases an average value seems to be the best compromise. The method of estimating PCB's described below attempts to take this into account. HCNS 06*5*1 4t L. M. IUtkou* Although most of the early reports oo the presence of PCB'j In samples indicated the Aroclor 1254 CLC pattern, more recent work by Kobn(an et at (1969) and he autliors laboratory has Indicated that a proportion of the specimens does contain the Aroclor 1260 CLC pattern. This apparent predominance of the Aroclor 1254 type might have been due to its availability as a standard and lack of tho Aroclor 1260 type in the early developmental stages of PCB methodology. The GLC profiles of a number of commercially available PCB mixtures were determined using the SE-G0/QF-1 column. These are shown in Figure 4 and Jnrii ate that the more highly chlorinated Fig. 3. Comrurium of ArocWs 1254 and 12t0. C]-'xt>atngrm A: 5 og. of Aroclor 1254; tho 14 motor peak ore number* 2 I to XIV as in chroma* tefram B ol Figure 1. Chromutopwm fi: 5 c;, oi Aroclor 1260; tho 1" [for peaks ire numbered 1 to XVII ttitc ' idv peaks cormpood to thooo In Aroclor 1254). GLC parameter* as tn Figure 1 biphenyls (Afodors 1254 and 1290) are easily detected with tho usual operating parameters. On the other hand, the compounds of the lower cnlorinatea mixtures (Aroclors 1221, 1232. and 1242) and the higher molecular weight mixtures (c.g., Aroclor 546 ) are less responsive and are, therefore, more likely to go undetected under normal operat ing conditions. Apparently, the lowered sensitivity is due to low chlo rine content in the first group and to high r.olecular weight with resultant long retention times in the second proup. In both cases, therefore, higher concentration would be a prerequisite to ensure detection under normal conditions. This, as well as the possibility of metabolism of the less highly chlorinated biphenyls (Koeman ct HONS 046532 i'. " ' l Polychlorob pheoyU 43 d. 1969), might explain the genera absence or rarity of these com* pounds in wildlife samples. It must not be overlooked, there 'ore, that in the presence of large mounts of the lower chlorinated bhihenyls (c.g., Arodors 1221,1232, t tt jnh i* 17 ad to - mual lower higher 'VHisive perat* dtlo..lrf with cases, * ensure uibOity 1 1MAK (t : > i. ; j i | 1 , { 'I Fig. 4. CLC profiler of tho more popular Aroclor mixture* under normal analy tical condition*. Koto: AlI peak number# correspond to thoao of Aroclor IBM. Cbomatograms: A - $ ng. of Aroclor 12x1. B + 3 og. of Aroclor 1 I23S, >C 3 ng. of Aroclor 1242, D 5 ng. of Aroclor 1294, E " 5 ng of Aroclor 1200, and F - 5 ng. of Arodor 5400. CLC parameter* aa in Figure 1 and 1242) and negligible quantities of the higher chlorinated bi* phenyls (e.g., Aroclors 1254 and 1290), different criteria would be necessary to detect PCB interference. To date, this condition has not boen enoountered In Canadian wildlife samples. \ MONS 066533 a** . m*U 44 L. M. !Ut*b nJfr Ai might be expected from t' e complex nature of the PCB mix* hire* aqd t^e chemical lnertne% of the individual compounds, die confirmation of iefentities of these materials is much more difficult than.wMijMferjlhocMorine pesticides. Thus, in the absence of a mass! s(kctnf|ler, confirmation* of the PCB's Is mainly by their GLC retention times1 and patterns on the mixed and polar phase columns in conjunction with their non-reactivities with the two general reagents used for the organochlorine pcstir ides. The prior separation of tiie P }B's from the organochlorine pesti cides by t|||kFl^^echni^i|||l^.so aids,In the Identification of the X. Estlmat on of PCB's Since the PCB's are themsclvrs toxic, attempts have been made to estimate the amounts present n samples. It should, however, be recognised that only rough estlm ites can be made since pure stan dards of the Individual compounds in the mixtures are to date unavail able. The situattajl resembles th t of toxaphene estimation, but la of ^greater complexity. Xocncan et cl (1009) based their PCB estimation on the peak of phenoclor DP6 having r, (relative retention time with dieldrin > 1 under his operating conditions) equal to 1.45. Riskmouch et of. (1969) estimated Ihe PCB s on the assumption that they have similar EC responses to p.y'-DDE and applied a factor to fit the assumed 54 percent chlorine content of the PCB's. Jtxsrs ef al (1909) re ported estimates as the sum of all the PCB components. The author's laboratory uses a method very similar to Koeman'i with the exception that an average based on two peaks is usually employed. Should the two values before averaging vary to any extent, additional estimations are made using other peaks, and all the results are averaged. In the case of a few samples, the PCB level was ob tained by averaging results from all the major peaks in the mixture; however, the use of peaks eight and ten only has been found to give equally satisfactory results. Note that results are reported as Being based on Aroclor 1254 or 1200 depending on the overall GLC t pattern. Peaks eight and ten were chosen because of their stability (see section on Identification). In a later section the above points are Illustrated by analysis of resin powder taken from a fish hatchery trough. Tne total PCB results obtained by the method described may be slightly higher than the actual values because the early emerging PCB peeics (mainly one, two, and three) are usually absent from the-chromatograms of sample extracts. However, this should not affect Ihe overall results greatly since these peaks represent minor compo nents In the 'Mixtures. Furthermore, there might be compensation, since no Attempt has bear made to apply recovery correction factors. t MOMS 066534 'il1k', I i i* t t{ * \ |i 4 Folychlor ibipheayb 41 The method of estimation de* ribed above appears to givttrratonsble estimate of PCB contain nation booed on commercial PCB mixtures. Eor absolute measurenent of PCB content it would b Decenary to prepare standards ft r the individual PCB components, end, even If tliesc were avails! Je, the detenninations would be complex and costly. Considering our present limited knowledge of total PCB toxicity and our even ' yeeter ignorance about the toxicity of the Individual components of he mixtures, it is doubtful whether such accurate detenninations wo dd be meaningful at this time. As stressed earlier, the most urgent r sed is for toxicological investigation in this area to indicate whether some of the PCB compounds are more toxle os have greater subkthal effect than others, as observed with the BHC isomers. At that point it would become essential to identify end estimate accurately t pacific individual compounds in the mixtures. XI, Separation of PCB's fron organophosphorus compounds by the Plot isil technique A limited amount of work has been carried out in our laboratory to determine whether the Florisi column technique is applicable to the separation of the PCB's from the organophosphorus compounds. Preliminary results on those tested (phorate, diazinon, ronnel, malathlon, parathion, methyl Trithion. ethion) Indicate that they are not eluted with the PCB's by hexane, and can therefore be separated. In comparison to the organochlor.no pesticides, the organophosphorus compounds generally require larger volumes of the ether-hexane mix ture for elution from the Florist! column, most likely because of their more polar nature. As with the organochlorine pesticides, the presence of PCB's in a sample extract will interfere with some of the more common orgenophosphites when the EC detection system Is used. However, with the use of e duel detector (EC + phosphate or thermionic) connected to a dual-pen recorder, the effluent from a single Inlection containing PCB's ana ornnophosphates can be split and differentiation made as shown in Figure 5. Like the organochlorine pesticides, the PCB's an not detected on the phosphate detector. Nevertheless, in the ab sence of n detector which is specific for phosphorus, die Florisi] col umn technique can be used to separate the PCB's from tbe organophosphetes to facilitate the analyses. XU. Residues of organochlorine pesticides end PCB's in Canadian wildlife Following the approaches outlined, the author's laboratory has analysed a number of Canadian wildlife specimens for residues of - -4. .. wg NCNS 066535 I I to IS 10 II 10 u MiftVtM Differentiation of Aroclor 1254 utd orginophotpltomt pettirtdei utb| , ' duaH detectors ad effluent *nlH. Chrftn>AtnvrMn A- orraoophcfphont, standard* pctllcide nblur* peak number* I 1.75 ng. phorate, I 5.0 ng. nf dlatinon, 3 o.O ng, of Ronnel, 4 " 13.0 ng. of maJithion, 5 5.0 ng. of paralhion, 4 * 150 ng, of meihyl Trithton, and 7 10.0 . ng. of ethton. Chromatogrim B: combination of organophotpl'orua mix* ture (chromatojram A) and 10 m. of Aroclor 1254 numbered 1 to XIV at In chromatogram B of Figure I (a compromise of the E.C. response b necessary to obtain maximum response on the phoiolionia detector). f * phosphorus detector and C electron capture detector. Column (fur percent OV-lOO/six percent OV-210 on 60/e0 mesh Chromoaorb W (AW)i other CLC parameters as in Figure 1 organochlorine pesticides and PCB's. Some of the data on aquatic birds from Western Canada are given in Table I. The inclusion of these tabulated results, which represent only a minor portion of the total number of samples annlvsca, is intended to illustrate the types of analytical problems created by the presence of PCB's, and to indi cate potential errors due to high levels of PCB's in samples. Referring to Table 1, some general remarks are in orders 1. Tjhe greater portion of the apparent DDE in samples is in fact DDE. This Is predictable from the comparative))' smaller amounts of apparent DDD and DDT. and has been verified by IIjC. & A large proportion of the apparent DDD in the samples Is due k to PCB's. However, DDD is present in some samples. I * MONS C66536 Polychtb -'cbtpbenyb 47 3. The effect of PCB contamina ion on DDT values U less marked than In the case of DDD, but ii Is stUl substantial. 4. The apparent HE values are in most cases due entirely to the pesticide. This is somewhat ii congruous with the rest of the data since peak four of the oomn ercial PCB mixtures does interfere with HE. However, this anor ialy may be explained on the basis that the PCD component givi ig rise to peak tour is degraded exteruively in the bird's metaooba processes. 5. Based on all of tire PCB est mates in wildlife samples noted to date, including those in Tabl I, the following general trends are evident: (a) Aquatic raptorial* generJly contain higher residue levels of pesticides and FCB's than oth :r wildlife species. (b) High levels of PCfis a*.- found only in the presence of high levels of organochlorlne pcstcides (the resin powder mentioned below Is an obvious exccptio)) but high levels of organocldorine pesticides are not necessarily iccompanicd by high levels of PCB's. (c) The most abundant PCB types detected are those which show Aroclor 1251 and 1260 pattc us and, therefore, fit well u Ith the proposed method of quantita ion. However, It should not he over looked that with the present i :'tiduo mctlrodology some other PCB* type compounds might go uncYtccted. 6. It is important to measure :ooisture and fat content of samples in conjunction with residue levels. Tliis allows fot repotting on a wet*, dry*-, or fat-weight batis. However, caution should be used when reporting residues in wildlife samples, for example, on a fat basis when the fat content is very low. Such figures can be inaccurate as well as highly misleading. The gas chromatogram of a typical sample sltowing the presence of PCB a is given in Figure 6. It indicates the Aroclor 1200 pattern and was obtained from a duckling found dead in the Toronto area in the summer of 1909. A 'good example of PCBs showing up In unexpected places oc curred recently. A wildlife biologist was faced with the problem of fish dying in a hatchery for no apparent reason. He scraped some of the resin powder from the hatchery trough and found, surprisingly, that experimental fish were susceptible to even a dilute solution of the powder. He submitted a sample of the resin powder to the author's laboratory, and upon analysis a typical CLC pattern of PCB's (Aroclor 1254 type) was obtained. This is shown In Figure 7. Using tire meth odology described earlier in this review, the sample was found to contain PCBs but no pesticides. Based on calculations from 11 indi vidual peaks of Aroclor 1134, the PCB concentration was estimated at 35 p.p.m (range 25 to 40). Using the average results based on peaks eight and ten only, the rCB estimate was 29 p.p.m This is MOMS 066537 it \ r )- r . 1 5 r 5 f MUNS Q 66S 33 \ l / t * I ,,'-DDT HE 1 Seecriptaoarof py* PCB* DDE? dria* (apparent) After u*u<w---' (apparent) After Brfetu (appareat) After ( Sample J area 6 Sample 2 area 0 Sample I arts 7 Sample 2 area 7 Sample 1 area 8 Semple 2 area 9 24 0 11.2 13.7 13.3 3.7 0.3 9.1 0 42 0.19 0.74 0.07 0.08 0.08 0.07 0.33 0.07 1.03 0.07 0 12 0.09 0.06 0.27 NO 1.70 0.03 N!> o.o. 10.03 81 0 104 04 0 49 02 9.18 0.30 0. OH 9.05 9.14 0.01 0.07 0.19 0.23 0.09 0.02 0.03 0.04, 0.CJ 57 77 09 33 IS 22 70 0.15 0.10 0.06 0.02 0.02 0.02 0.03 0.12 Trac^ 0.06 Traea* Trace* 0 01 0.01 ss -- 100 -- -- 52 44 3.41 1.93 1.09 0.57 1.38 0.53 9.4? \ it These specimens are from waters Canada (Vbbmcrk and Ititholds 1970). These are "Before PCBFloridl separation" values and ar > likely to iwhide PCB contribotieae. The "After" value* (which do oot chance significantly fm the "Before" valors) are not gi~en ><eemawe the PCB peak fire (the DD&mtcrfering peak) of Arodor 1254 or I2G0 is oot separated from Dl>fc by this ntethod, ba% as diseased earlier, the PCB contributions to the DDE rahM sro exported to be relatively small compared to the contributions to DDD or DDT. These are "Before" values. The "After" vahws (not given, are not eicntficaatly different from the "Before" valwa anee there is as direct PCB interference. However, in the presence of high levels of DDE, low levels of dieMria will be masked. In such caw* a separation (eg., PCB-Ktorisd techoique) is tie reawry, and the "After" raise would he reported ae the dkUria level. % - After Tfclne ^ jqq ^ infiau* the percentage of the total apparent pesticide level that waa actually due to the pmtkide Before value Vahw* (p.p.m.) based on peaks eight and tau of Aroder 1234. /HD -- none detected <0.0001 p.pun. Trace <0.001 p-p-ia. 6* * i i .i MCNS 0 6 6 5 3 9 Fig. 8. T)pUaJ sample Indicating tl e presents of PCB's of the Aroclor lt60 type la deciding from the Toronto area. Chromatogram A: 5 ng. of Aroclor 1260 numbered I to XVII as In chromatogram B ill Figure 9. , Chromatogram V; the equfv Jent of 0.35 mg. of duck sample (hereof , portion of Florhfl split). Nile the DDE contribution to FCB peak V. CLC parameters as to Figure I .. si. * i* F|g. 7. Resta powder attract indicating the presence of PCB'a of the Arcelor 1854 type. Chromatogram Ai 5 ng. of Aroclor 1854 numbered 1 to XIV at to chromatogram f of Figure 1. Chromatogram B: The equivalent of 0.08 mg. of resin powder extract (hetsne portion of FloritU spilt), numbered I to XIV as to Aroclor 1854. CLC parameters as to Figure 1 1 '*> < ''li: ? *0*s 066540 Polychli roblphenyla II an interesting example of I'CB': causing contamination as a direct result of Industrial application. With regard to tne Florlsil t.-chnlque for the separation of PCB's from pestiddes, one important nodiucation to the original method ( Reynold* 1009) it now used rcutincly In the wildlife studies. Origi nally the scheme was designed o accommodate simultaneously both the cleanup of the sample extract and the separation of PCB's. How ever, as pointed out earlier in his review, not all samples contain PCB's. Consequently, for genera samples, to avoid unnecessary split ting of all sample extracts (eact split doubles the number of eluates to be chromatographed), a non mi cleanup including a Florisil col umn, but no differential elution, is conducted. The Florisil-PCB sepa ration is then carried out on tlw clcaned-up extract, if it is required. Under these conditions smaller columns have been found to be as efficient ns the larger columns u ed in the original method, and there is a substantial saving in the amount of reagents required. Details of the modified procedure are as Allows: A glut column (44 cm. X I cm. .d. hiving 50-ml. reservoir on top) is picked with 30 cm. (--10.7 g.) )i Florisil (60/100 mesh, Floridin Co., pesticide grade, stored at 130X. mtil ready for use) and topped with a tem. layer of anhydrous Ka*SO. The sample extract of about 5 ml. '(n-hexane) Is added to the cotimo. The HrU elution which removes PCB's, heptachlor, aldrin, and DDE, is effected with 00 ml. of n*hcaane iNaitograde, MaUiuckrodl). The u;nj..lng pc;Ucidcs are then eluted with 40 ml. of 50 percent ethyl eth-ir in hexane. Note that In filliiig the coluim, live Florisil is packed down by gentle tapping, and the sample extract <* added to a dry column, if., no pre* watting. After the first elution the receiver is changed as soon as Ute last of the hexane meets the Na,SO. It should be noted also that with one batch of pesticide grade Moristl, tome PCS peaks (two, sta, nine, II) showed partial elution in the second eluate. These, however, are com paratively minor peaks and do not interfere with any of the common or^anoeworine pesticides. As there are differences in batches of Florisil, the procedure has to be cheeked out fully prior to application to actual samples. Modifications of volumes, etc., may be necessary to effect the desired separation. It is likely that adsorbents other than Florbil, .g., afffee g4i or alumku, will give similar fepar^tion of PCB's from pesticides but these have not been investigated to date. Acknowledgments Work in connection with this review was supported by funds from the Pesticide Section, Canadian Wildlife Service, Ottawa, and from the Province of Ontario through tho Department of Trade and Devel opment. The writer gratefully acknowledges helpful suggestions of Dr. S. C. Add, Director of Organic Chemistry Department, Ontario HONS 066541 u 41 I L. M IlUrmcta* Research Foundation in prepar ng this review lor publication. Hm technical assistance ol Terry Co )per and Sandra Taaacs ii gratefully acknowledged. T^b EL Chemical dcti^netl m *f peaflcidct mentioned tn ten PeiitMi Chmieil atm* aldrin 1,2,9,4,10,10-be tehlor-l,4,4i(5,8,Bi-bitbydrol,4-md, e*-ft,S-dimct lanonaphthakDe p,p'-DDD (TDE) 2,2-bil(j>-h)or6 riMBylH.l-dkhloroetbane py.DDfi . l,I-dfehloro*2,2 bi(p-*hloropbcnyl)*thyleD >,^ODT; Jl , ULljl'triebiqt^S &bte(p-eh)oropbefiyl)etba&e aiulaoa :(l' n MhOEdietlimM ,teopfopyM-melbyi'6pyriw>dyl)pb<i "r> pbovothkliW dhldria 1,2,9,4,10,10-be :achloro*6,7-poxy-l,4,4a>5,a,7,l,8e'Oete- tibioo bydro-l,4*m ),(xo>5,S-dimethtnoQephthxlene O,0,O',OMclrai ihyl-,5'metliyltM biapboephorodithioeU hepUchlor l,4,M7,9,b-bf| taeblorO`3a,4,7,7e>tetrfthydro>4,7*mdO' methannind* <e bspUcblor epoxide l,4,5,C,7,S,W*ej teehloro>2,3-epoxy-3e,4,7,7e-t(rehydr>4, 7*mthanftiix an* Ui*Um (vBHC) -r-l,2,3,4,A,0he eebloroeydohexeDe malathioo il,^bii(ihos/tarbonyl)ctliyl)0,0-dimcthyl pboepborodi* IbfoaM Mlhji Tritbfea O,0-dimtfiyl r-(p-cftloropncnylU>ioj methyl phoapborodt- tbioete paraibtoa^ p'bowratlo (Thinet) Rooaane"i 0,0-diethyl 0-p nitrophrnvl phoepborotbioete 0,0-dicthyl 5-u liiyltbio) methyl phoepboroditbioeto dimethyl 2,4,&'trJeblon>phcnyl phoephorothionele Summary . Since 1066, PCB's have been detected in fish and wildlife, espe cially In Europe and North America. Contamination from industrial wastes via the water ways appears to be the main source of PCBs In aquatic species. The PCBa have many important industrial uses but are not utilized as pesticides. Due to their similarities in structure and properties to the orgonochlorine pesticides, the PCB*$ tend to interfere with accurate GLC-EC determination of organochlorine and many orgaoophosphonit pesti cides. In addition to their interference with pesticide residue analysis the RGB's are themselves toxic so that their quantitation (s desirable. A ipethqd for recognising the presence of PCBs In samples, sep arating them from pesticides, and determining the pesticide levels accurately is described. Simultaneously an estimate of total PCB con tent `is afforded '~T' MOWS 066542 PolycUotcbipheByb The residue data of tome Oi tadian aquatic bird* indicate that high levels of PCB's are found only in the presence of correspondingly high levels of the DDT pesticide group. On the other hand, high organochlorinc pesticide levels are not necessarily accompanied by high PCB content. Further research on toilcity and sublcthal effects of PCB mixtures and their individual components is required. If particular components are shown to be more toxic than < thers, refining of the PCB quantita tion methods will be necessary Rdsumd* Analyse de rdsfdus de jesHcidci en presence de diphenyls polychlords Dcpuis 1906 dcs diphemls p lychlords (PCBs) ont dtd detect4* dans lea polsions et lo gibier, MrticuWrcment en Europe et on Amlrique du Nord. II apparaft jue la presence dc PCBs dans la faune aquatique soit due principa ement & la contamination dcs cows d'eau par dcs dcchcts industriel. Lei PCBs soot utilises pour un grand nombre de proeddds lndu>triels important!, mais lls nc sont pas appliques comine pesticides. Etirnt d^nud que In structure et les pronrict^s des PCBs sont semblablet I celles des pesticides organochlords, ils ont tendance I fausscr la determination etacte des substances organochlorees et organophosphorees lors d'analyscs par chromatographie en phase gazeuse via capture delectrons. En plus dc leur interference lors de l'analysc dcs rdsidus des pesticides, Its PCBs sont eux-mdmei des oon '* `oxiques dont la determination quantitative rst souhaitable. ent article conticnt unc mdthodc qul permet de reconnaltre la presence des PCBs dans dcs dchantillons et, en mdme temps, de les sdparer des pesticides dont le taux de rdsidus peut ttre eiactement ddtermtnd. La methode permet dgalcrocnt une estimation du contenu total en PCB*. Des rdsultats d'analyscs portant sur des oiscaux aquatiques cansdiens indlquent qu'un taux dlevd de PCBs est lid 4 un taux dlevd de rdsidus de pesticides du groupe DDT. D'autre part, des taux dlevd* de rdsidus dc substances organochlordrs ne sont pas ndcdssairement aocompagnds d'un taux dlevd de PCBs. Des recherches suppldmcntaires sur la toxicitd et les effets sub* Idtaux des mdlanges de PCBs et de leurs composes individuels seront ndcdssalres. Si on peut ddmontrer que des substances particuildre* sont plus toxiques que dnutres, on aura besoln d'une mdthode plus spdcifique permettant fanalyse individuelle des PCBs. * Tradult par H. CoasaOiu*. MCNS 066543 L ,M. Kitmolm ,I ZUMtr OIMlfuniltf* Die Analyte von PflnnzeniihutzmlttelRtlckjUodett In Cegenwart / . von Polychtordiphenylen Seit 1900 werden Polychlordiphenyle (PCBStoffe) In Fitchen und Wlldtieren, bcsondcrs In Europa und Nord.imcrika, oachgewinen. ! Die Venchmutzung der Wnsfsrwege durch Industxieabfille scheint dabel die Hauphirsache von PCB-RilcbUodcn in dcr aqoatischcn Fauna ru scln. Die PCB*Slo (e werden fiir fine Anzahl wichtiger Industrieprozessf gcbraucht, ai; Scbkdlingsbekiimpfungsmittc] werden tie fedocn nicht eingcsctzt. Da die PCB*Stoffe in Ihrer Struktur und in ihren Eigcntehaften den chloricrlen Kohlenwasse itofMnsektizidea sehr Ihnlich liod, IcOhnen lie die gaschromatogi iphische Bcstimmung mil Elektronen- einfanjg-Detckter von chloricr cn Kohlenwrmerstonen und mancber Phpsphoriiiurecstcr verfBlsche*!. Die PCB-Stoffe kttnoen nicht nur RUckstandsanalyscn beointrKcl tigcn, tondem tie lind selbst toxisrhe Substanzcn, dercn Nnchweis etch aufdrlngt. Ira vorlicgendcn Artikcl ulrd cine Nfethode beschrlebcn, die ex crtaubt, dot Vorhandenscln vo i PCB-Stoffcn in Probcn oachzmveisfn, tie von den SchlidlingsbekH. npfungsmittcln abzutrennen und die RUckstlnde dcr letztem genau zu bestinmico. Dariibcr hinous gestattet die Mcthode, den To'.algchaH an PCB-Stoffen abzuschtttzea Rtickstttidsduten clniccr k.tmd^che* wriem darnuf hln, dast hohe PCB-konr.ctitraHonen nur In Cegenwart hoher RUckstXn^c von Substanzcn der DDT-Cruppe auftreten. AndcrsdU lind hohe RUckstttndc von chloricrtcn KohlcnwasserstofiMroektizidco nicht unbedinct von hohen PCB-Konzentralionc-n beglcitet. Weitcre ifntersuclmngen tiber die Toxizftiit und die sublethalc Wirkungvon PCB-Mischungcn und ihrer individucllen Kompopenten tind angezeigt. Sollte rs rich dabei ergebrn. riass gewissc Einzelstode torUcber slnd als andere, wird Hne Verfeinening der Nachwcis- und I Bestimmungsmethoden fiir PCB-Stoffe notwendig. ' Reference! Bboce, 6. S-: Insecticidal surface coatinp. Soap 4 Saolt. Chem. t4 (2),135 n (1W8). 11 Bnowx, R. M.: On the toxicity of the "Aroclor". Chemist Analyst 96, 93 (19-17). , Buamcx, p. E.. E. J. Harms. H. J.Diam, T. M. WaubivJ. Skka, and D. Count: The accumulation of DDT In lake tro'it and the effect oo reprodue* .I tho. Tran*. Amer. Fisheries Soe. 93 (2), 127 (1964). . Rums, ]. A.t Cm chromatography for pesticide re due analytic some practical > i aiptcti.). Assoc. Official Anal. Chemists 42, 1037 (1965). CRAU, A. S. Y.i Derivative formation for the c?r./>rmation of tndoiulfan by CM chromatography. J. Assoc. Official Anal. Chemist* 52, 1240 (1969). HONS 066544 Polychlorobi benyJa n ,----, and W. P. Cocmmkii Cyclodien chemistry. III. Derivative for-*.eite for the identification ef bMichlor etide, cb-rModtM, (rinMkicniie^ die)drin, end endria pesticide retidu by gas chromatography. J. *ijioc Official Anel. Chemists 88, 1220 (1000 . Cochkavx, W# F., end A. 8. Y. Ckau: Noti on gai chromatographic kfrntiftc.irion of beptachlor pesticide residues by terivative formation. J. Assoc. 0ciai Anal. Chemists 81, 1867 (1066). Dueou, 1C P.r Combined effect* of pestle de*. In; Current views on pestMdea, p. 65. Symposium sponsored by Fool end Drug Directorate, Dcpw-r-.-nf pt Nation*! Health end Welfare, Otb we, 1068; reprinted from Can. Med Assoe. 1.100 (No. 4) (Jen. 05, 1060). Dotty, J. R., end N. Wovot Residues of organocblorine Insectleidei end tbetr metabolites In soils in the Atlantic pro foces of Canada. J. Acs. Food Chem. 15, 457 (1067). . Chekmdum, L, M. R. Mayers, A. R. (semi: The systemic effects resulting from exposure to oertaln chlorinated hydrocarbon!. J. Ind. Ilyg. Totted. !, 80 (1030). Cuntuck, F. A.; Instnimentation in pest* ide residue dctenninaHont. Adv. Pest Control Research 5, 191 (1008). , Hamminci, J. If., P. 8. Hals, and D. J. tt*vmi.vi Tha Identification and detir* m(nation ef chlorinated pesticides mid ws. Anslyst 90, 049 (1005). Hamusox. R. B.'i Residues to wildlife wl*b special reference to endfin. J. $d. Food Act. 17, 10 (1060). Hacnv, J. J., and D. W. Avocrson: C lorinated hydrocarbons and eci!*hdl changes In raptorial and fish-eating lrd#. Science 108, 871 (100S1. Holden, A. Vm ana K. Marshes*: The ex itotlon of surface waters and sewage effluents for onganochlorine peak-idea. ]. Froc. Imt. Sewage Purif. p. 895 (1000). ----- - Organochlortoe pesticides to seals and porpoises. Nature 810, 1874 (1067). HoLMtt, D. C., J. II. Simmons, and J. O'Ci. Tatton-. Chlorinated hydrocarbons to British wildlife. Nature 816, 827 (19M7), Hornsteim, I., and W. N. Sullivan.- Th-i role of chlorinated polyphenyls to Improving lindane residues. J. Econ. Entomol. 46, 937 (1053). Jensen, S.5 Report of a new chemical hazard. New Scientist 38, 618 (I960). --------- , A. C. Johnels, M. Otssov, and C. OrrsRLCfti DDT and BCR to marine an(mall from Swedish waters. Nature t24, 847 (1060). --------- , and C. WimusiK: Swedish report at the OECD pesticide conference on Unintended residues in the environment'* (1067). Jones, J. W., and H. S. Alms*: An acneform dermatergosis. Arch. Dermatol. Sypbdol. 33, 1082 (1036). Kixix, A. K,, said J. O. Watts: Separation and measurement of perthane, DDD (TDE), end DDT to leafy vegetables by electron capture gas chroma tography. J. Assoc. OSctal Anal. Chemists 47, 311 (1004). Kokman, J. H., A. A. C. Oskamp, J. Vick, E. Brouwer, J. Booth, F. Zwadt, E. v. d. Bmmk, and H- van Cxviwren: Mailed. fUfkt'fecutMt Larufhouto- watentchappen Cent. 38, 641 (1067). Koeman, J. H., M. C. ten Noeveii dk Bruu, and R. H. m Vo: Chlorinated biphenyls In Ish, mussels and blrdis from tbe River Rhine and tbe Nether lands epastaiarea. Nature 881, 1186 (1969), Lindquist, A. w., A. H. Madden, II. C. Wilson, and E. F. Kkikjko: DDT as a residual-type treatment for control of house files. J. Econ. EntonoL 38, 857 (IMS?. McLauokum, J., Jr., J. P. Marlsac, M. J. Vekkett, M. K. Mutcmlo. ind O. C. fmivucN: The Inaction of chemicals Into tho yoUc sec of frble S^^rto to incubation u a toxicity lest. Toxicol. Applied AanuMl. 1 HONS 066545 t < i i r L. M. REYNOLD* Mum,W.s hdMlacle d> u>;ci in animals exposed to oomntrcbl Bt|rj diphenyl. U. 8. Public I selth Reports 89, 10S9 (1944). IfaiMftfo Co..* The Aroclor compound), p. 17 (1905). -->-- Aroclor plasticiser*. Tech. Bull. O/PL-300 (1907). Ooadcmucx, M., and E. B. Wattless: tdcntJSc.itlon and quantitative esfjt milon of aldrin residues in tim pretence of Lvrrfering materials in iam| m f plant and animal origin, |. Auoc. Official Anil. Chemists 51.1264 ()06i). Psaull, D. B.t Pesticidc-imiuoed entvme br< sfcdown of steroids in birds. ; 'atvtt * 810, 505 (1907). ' ' FKnnino, C. H.: Physical c laracteriatic* and commercial possibilities of hlori . nated biphenyl, ind- Er. J. Chem. 82, 11 SO (1930). lUicurrc, P. A.': Decrease to eggshell xveizht to certain birds of prey. Wattnr I . 815, 806 (1907). Rschcl, W.t Personal communication (1969V FUtkolm, L M.: Canadian wildlife service reports (Unpublished data) I960, *, 1909). | --* Pesticide residue aalysls In the presence of PCB's. 4th Annul W, Can. Seminar on Prsticile Residue Anx' >ii. Winnipeg, June 8-4 (19 9 a). --------- Polychlorohinhenyls PCR's) and their interference with pesticide r sfcNr analysis. Bull. Environ. Contamination Tosieol. 4, 120 (I960 b). RstnnoucH, R. W.. p. ftis an, S. C. lltn'UN. D. B. Pkakall, and I. N Kimvem; Polychlorinated biphenyts in ibe global eco-system, Nature tH, 1090 (i960). -------- , P. RacMi:. and II. 8. Olcott: Current progress b the detemu letlan of the polvchlorlnated .'toheuvls. Bull. Environ. Cool snUnation Toskul 4, 198 (lw9). ' Ronixaox, J.j Residues of oryanoclvlortoe inwtV.cldrt to dead birds in the t olled Kingdom. J. Chetn. Ind., p. 1974 < 196T >. RoeunN, J.: A simple cnncpnirntion-erll techni y<e for determining small of halide ions and Its use lit the detemrmtion of residues of tcanoch orinc pesticides. Analyst 90, 467 (1965). Samuel, B. b.. and H. K. Hooctt: Insecticide trreenlng methods for nrcan< chto rine and ortaoophosphate insecticides to foods aod feeds. Residue Reriews 17, 35 (1087). Sans. W. W.t Multiple insecticide residue determination using column chron atof- mphy, chemical conversion, and gat liq-:;d chromatography. 1. Agr. Food Chem. 15, 192 (1967). * Schmidt, H., and C. Schulte: Patent for manufacture of pentachlorobipliemi Ann. 807, 338 (1801). ' * Schwamtz, l>: An outbreak of halowax ante *"cable rash") among electricians. ' f. Amer. Med. Assoc. 188,153 (1943 V . - , and F. A. Barlow: ChJoracae from cutting oils. Public Health fieri. 87, 1747 (1942). --------- , and S. M. Peck: Occupational acne. N. Y. State Med. J. 43, 1711 .(1943). [ScuacKTYit, M. S., S. R. Solowav, R. A. Have*. and II. L. Hallan: Cnlnrlmrtrtr ' I determination of DDT. Ind. Enc. Che:.'... Anal. ed. 17, 704 (19151. Srsonaa, K-: Calcium In reproductive pnyilc*l.->" . New York: Rclahoid (1967). SttCkkl, L.j Wisconsin bearings. Science 163, 551 (1909). j Suuuivan, W. N,, and 1. Hornctm*: CMoriiuud polyphenyls to improve Itada* residues. J. Econ. Entomol. 40, 158 (1953 . Thompson, J. F., A. C. Waloe, and R. F. Mcheman: Study of the performance of gas chromatographic columns under >evete Infection loading. J. Assoc Official Anal. Chemists 62, 1251 (1969 a\ * - -......-- -- Evaluation of eight chromatographic columns for ehlori aated pesticides. J. Assoc. Official Ana). Chemists 68, 1803 (1909 b). '*' HONS 066546 t * ,^i^'^llfc^^,.^.|,I H" TmA C. H., W. N. Suluvan, And I HonNSvan*: A afevaporaftoa ` rates dm toxicity to bouse flki oi Undane And Undine>chlertaated pifypWyl deposits. I. Eeoo. Entomol. 49, V i (1953). V. 5. rood md Dnr Adminteratt*; Pesticide Analytical Manual, VoL 1 tod II, Nviicd annually. Washington D.C. (IBM). Van 'Tot, N.j Improvement o( tlte residual toxicity of DDT solutions by the edditloo of coumArone resin. B ill. Entomol. Research 43, 413 (list). VtsMsm, K., end L- M. Ravnoumi 3rganechlortne residues in aquatic birds in the Canadian Prairie Provinces. ( n preparation, 1970). Wastm, R. M., M. Levins, and A. 11 Cosset: Effect of chlorinatad insecticides on steroid metabolism. In: Che nice) fallout, proc. 1st Rochester Coof. on Bovlronmental Toxicity. Vniv. of Rochester (1909). WiDMaac, C.: IUPAC commission on methods of pesticide residue analysis. I. Asaoc. Official Anal. Chemists 0,1009 (1907). Wlkscot, W. W,, and f. A. Biku Derfvatiaation of dieldrin and andrin for confirmation of residua identity I. Assoc. Official Anal. Chemists 12, 1277 (1909). Zweiq, C. (ad.): Analytical mdtods for pesticide*, plant growth regulators and food additives, voi. 1, p. 1. . lew Yorx: Academio Press (1903). HONS 066547