Harriss, R. C., Nature 219, 54 and 55 ( I 965). Hosohara, K., Nippon KugukuZusshi82, 1107 and 1108 (1961). Hosohara,K., Kozuma, H., Kawasaki, K., Tsuruta, T., Nippon Kagaku Znsslii 82,1479 and 1480 (1961). Irukayama, K., Adc. Water Pollut. Res. (Munich) 3, 153-80 (1967). Kurland, T. L., Faro, S. N., Siedler, H., World Neurol. 1, 370-91 (1960). Raeder, M. G., Snzkvik, E., Kongl. Videnk. Selsk. Forhandl. 13, 169-72 (1941).
Stock, A,Cucuel, F., ’Yururwissen22,39943 (1 334.). ‘Turekian, K.K., ’Weclepohl, H., Buli. G d . .5’hc. Amer. 72, 178-92 (1961). U. S . Bureau of Mines, “Minerals Yearbook; 1966,” Vols. I, 11. “Minerals and Fuels,” 1967. Williston, Samuel H., J. Geophys. Res. 73,7051-55 (1968).
Receiwdfor reciew January 15,1970. AcceptedJune 18,1970.
COM M U N I CAT IONS
Recovery of DDT and Dieldrin from Tissues of Coturnix Japonica Stepwise During Residue Analysis Richard M. Prouty and Eugene Cromartie Bureau of Sport Fisheries and Wildlife, Patuxent Wildlife Research Center, Laurel, Maryland 20810
Recoveries were measured stepwise during a 5-part analytical procedure by counting carbon-14 labeled p,p’DDT or dieldrin. The greatest losses occurred during concentration of eluates from Florisil columns or zones from thin-layer plates and were probably due to codistillation. Nonetheless! overall recoveries of 70-94 are possible. H
F
or several years this laboratory has been analyzing tissues of field-collected bald eagles and other bird specimens for chlorinated hydrocarbon residues. A 5-part analytical procedure (Reichel, Lamont, et al., 1969) has been developed for isolating these residues from tissue lipids. Stepwise, the wet tissues are chopped, mixed with sodium sulfate, and extracted for 6 hr. in Soxhlet with petroleum ether (30’-60’ C., b.p.). The pesticide residues are grossly partitioned from the tissue lipids in hexane (60’-70’ C., b.p.) by acetonitrile extraction. The remaining lipids are removed by Florisil column chromatography with elution of the residues by a 3:l mixture of hexane:benzene. The residue mixture is separated into zones on a thin-layer plate (silica gel G, developing solvent-2x ethyl ether in hexane) with elution from each zone by boiling benzene (Mulhern, 1968) followed by analysis of the zonal eluates by gas chromatography (3 OVglass 17 on 100-120 mesh Gas Chrom Q in 6-ft. X column, operated at 197” C., 100 ml. N2/min.gas flow). To measure the percentage recovery for this analytical procedure, laboratory trials were conducted using tissues containing biologically incorporated carbon-14 labeled p,p ’DDT or dieldrin. Experimental
Fifty-pc. p,p ’-DDT-C’* (ring labeled) (New England Nuclear Corp.) were diluted with 90 mg. of the unlabeled compound in corn oil and fed by capsule at the rate of 3 mg. DDT per bird per day to three Coturnix quail for 10 days. Fiftypc. dieldrin-CI4 (Amersham-Searle Corp.) were diluted with 768 Environmental Science & Technolog!
150 mg. of the unlabeled compound in corn oil and fed by capsule at the rate of 4.25 pg. dieldrin per bird per day to three Coturnix quail. This dosage proved fatal hithin 7’days to these and four replacements. Both living and dead birds were used to provide tissues for analysis. To estimate the radioactivity in the tissues before Soxhlet extraction, two 100-mg. aliquots of each sample were solubilized in perchloric acid and hydrogen peroxide and counted according to the method of Mahin and Lofberg (1966). The entire sample of each tissue was then extracted and recoveries determined after each step in the following manner: for DDT, extraction, partition, and column elution solvents were evaporated to dryness, the residue redissolved in benzene and two aliquots, each equivalent to 100 mg. of sample before extraction, were taken for counting. For dieldrin, extraction and partition solvents were evaporated to dryness, the residue redissolved in hexane and aliquots taken for counting. Column eluates were concentrated to about 10 ml., transferred to graduated tubes, and made up to volume with hexane before taking aliquots for counting. For separation of D D T and metabolites, the thin-layer plates were divided into four zones, the first zone extended to include the origin line. For dieldrin the plates were divided into five zones, four plus the origin line. Each zone eluate was counted separately and the compounds identified by gas chromatography. Results and Discussion
The percentage recoveries of DDT and metabolites are given in Table I. By use of solubilized tissues as a reference base, petroleum ether gave complete extraction in Soxhlet of radioactivity from liver and brain tissue and nearly complete extraction from heart and breast muscle. The overly high recoveries from brain tissues after extraction and partition reflect the difficulties in sampling small-volume, high-activity extracts. Residues of unextracted radioactivity in the muscular tissues may have been compounds in the protein-bound lipids which resist extraction by nonpolar solvents, or water-soluble
Musc9e Liver Heart Brain Brain Liver Heart Breast muscle Shanks and tot.\
h
,
Aveiage of b o diquots.
* Not determined
metabolites, although these have been previously reported only in liver and feces (Abou-Donia and Mcnzel, 1968). T'hc lower recoveries obtained after Florid and thin-layer chromatography illustrate the losses due to co~stiliationthat can occur in virtually fat-free samples evaporated to near or cornplete dryness under an air stream (Burke, Mills, et 01.. 1966). Scanning of the zone eluates by gas c ~ r o m a t o g r a p ~revealed y p,p'-DDT as the principal compound present, with lesser amounts of p,p'-DDD and DDE. The percentage recoveries for dieldrin are given in Tabk I. Depending on the tissue, duplicate variability in the counts of samples solubilized with perchloric acid and hydrogen peioxide rdngecl from 7-30%, and counts in an aliquot ofSo,xhlet extract exceeded those in an equiVahK weight of solubilized tissue by lO-lOS%. Use of a quaternary base (Soluene) as ;t solubilidng agent did not improve recoveries. These resuits suggest that dieldrin was partially volatilized and lost ituring solubilization; thus. all stepw'se recoveries are related in Table I to courits in the Soxhkt extracts rather than to L!OW in solubilized tissues. As with DDT, mine dieldrin was lost by
Sydney C. Barton and Nenr) G . MsAdie Ontario Research Foundation, Shcridan Park, Oiitario. Canada
A method for reducing blank effects in the collection of HBO? aerosol on glass fiber filters has been developed. 'Treatment with hot HzSOI foliowed by thorough rinsing permits the filters to be used to collect microgram amounts of H&04 aerosol without any significant irreversible lass of H$Oc on the filter.
G
lass fiber filters combine the desirable features of high retention efficiency of submicron particles with high flow rates, and so have found widespread use in the collection of particulate samples in air pollution studies. Such filters were used extensively for the collection of HnSor aerosol until i t
was shown (La: nn3 Wagman, 1966) thai atniospheric sulfur dioxide could be oxldired catalytically on rhe glass surface and thus sei iotsl? interfere with the deternunation of prevailing HSO, aerosol levels. Dilticulties encountered in allowing for blank efFects when .tnaljzing f x H SOt aerowl collectcadon gla5s fiber filters have also been discussed Druboih, Tachman, el id. (1967)report that inany brands of thcse filter5 containeil excessively high amounts of sullblsr tnd have de\crhed extraction proaxfurs bq which thew .,in be reduced. Howewr, Scaringelli and Rehme (1969) w c l r h r t the rcsidual alkali content of the filter can result in s d f w I C acid losses of 0.4 to 7 8 pg per cm. 2 of filter. Thus. there c a n ht. considerable vanation in the chemical propertie.; oi' plais f i k r tilters. anti t low ~ t t ~ mmt ~ ~ j ~ n be given to hEanl, effects when dcaliq; with a t ~ o ~ wnples contcimng micr
