763

Journal of Food Protection, Vol. 63, No. 6, 2000, Pages 763±767 Copyright ᮊ, International Association for Food Protection

Chlordane and Residues following Cooking of Treated Channel Cat®sh Fillets

C. R. SANTERRE,1* R. INGRAM,2 D. H. XU,3 G. W. LEWIS,4 AND L. G. LANE2

1Department of Foods and Nutrition, Purdue University, West Lafayette, Indiana 47907-1264; 2Mississippi State Chemical Laboratory, Mississippi State, Mississippi 39762; 3Department of Fisheries and Allied Aquacultures, Auburn University, Auburn, Alabama 36849; and 4Warnell School of Forest Resources, University of Georgia, Athens, Georgia 30602, USA

MS 99-215: Received 28 July 1999/Accepted 17 December 1999 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/63/6/763/1673844/0362-028x-63_6_763.pdf by guest on 27 September 2021

ABSTRACT

The reduction in residues of chlordane and toxaphene following cooking (frying, baking, and smoking) of ®llets obtained from treated Channel cat®sh (Ictalurus punctatus) was determined. On average, cooking reduced moisture content by 17% and increased fat content by 28 to 274%. Frying reduced chlordane residues by 56 to 86% on a dry basis (db) or 84 to 92% on a percent fat basis (fb) when raw ®llets were compared to cooked ®llets. Baking and smoking reduced chlordane signi®cantly less (P Ͻ 0.05) than frying with reductions in residues of 12% and 9% (db) or 30% and 33% (fb), respectively. Frying reduced toxaphene residues by 40 to 49% (db) or 65 to 77% (fb), while baking and smoking reduced toxaphene by 35% and 24% (db) or 51% and 59% (fb), respectively.

Numerous reports (12, 27) have documented the pres- studies previously discussed, that residues in wild ®sh may ence of organochlorine residues in wild ®sh including frequently exceed the FDA guidelines. chlordane (5±8, 15) and toxaphene (19). Average total The reduction in food contaminants during preparation chlordane residues in wild ®sh have been reported as high and cooking have been reported (17, 22, 23, 34). More as 6.69 ppm wet basis for wild ®sh collected in a national speci®cally, the reduction in ®sh contaminants during food survey during 1978 to 1979 (25) and less than 0.07 ppm preparation has been investigated for the past 30 years (1± in a 1987 sampling (8); however, typical average levels are 4, 13, 14, 16, 20, 21, 24, 28±33, 35, 36). Even though demonstrated by other reports that indicate levels in ®n®sh researchers have reported mixed results, it is clear from from Maryland waters at 0.12 ppm (7); carp from the Upper these studies, that preparation and cooking signi®cantly re- Steele Bayou, Mississippi at 0.15 ppm (12); carp and bass duce residues in ®sh. from Tuttle Creek Lake, Kansas at 0.1 and 0.092 ppm (5), The objective of this study was to determine the in¯u- respectively; Channel cat®sh from the lower Mississippi ence of frying, smoking, and baking on residues of chlor- River at 0.17 ppm (15); and Channel cat®sh from the Mis- dane and toxaphene in ®llets from treated Channel cat®sh. souri River at 0.116 ppm (6). Other researchers have re- MATERIALS AND METHODS ported average chlordane residues in farm-raised Channel cat®sh of less than 0.045 ppm (26) and 0.10 ppm (18). Channel cat®sh were raised at a density of 18 ®sh (15 kg) Recently, toxaphene residues have been reported by the in each tank (tank volume was 10.22 m3; water was exchanged Food and Drug Administration (FDA) (11) in the Total Diet ®ve times during the study) with two tanks for each residue. Fish Study and in infant foods sampled during 1998. Average were fed a 32% protein, ¯oating cat®sh feed (Alabama Farmers toxaphene residues in wild ®sh have been reported as high Cooperative IMC, Decatur, Ala.) for a period of 25 days at the as 21 ppm for wild ®sh collected in a national survey dur- rate of 1% body weight and then at a rate of 0.5% for next 13 ing 1980 to 1981 (27); however, typical levels are dem- days in order to maintain water quality. Feed composition was 32% protein, 2.5% fat, 6.0% ®ber, and 12% moisture. Chlordane onstrated by other reports that indicate average levels in (technical grade, Ultra Scienti®c, Inc., No. Kingstown, R.I.) and Largemouth bass and Channel cat®sh from Lake Provi- toxaphene (technical grade, Ultra Scienti®c, Inc.) were incorpo- dence, Louisiana at 8.96 and 2.27 ppm (19), respectively; rated into feed at 2 mg kgϪ1. Thoroughly ground feed was mixed in Largemouth bass from Lake Bruin, Louisiana at 3.58 with chlordane or toxaphene and made into pellets using 3% car- ppm; and carp from the Upper Steele Bayou, Mississippi boxymethyl cellulose as a feed binder. The total amount of treated at 3.06 ppm (12). feed consumed by the ®sh during 38 days was 5.4 kg. A group The FDA (10) has established action limits for total of ®sh (control) were fed commercial feed with no added residues. chlordane and toxaphene residues in the raw edible portion At harvest, 24 ®sh for each organochlorine treatment were of ®sh at 0.3 and 0 ppm, respectively. It is evident from collected and divided into four groups of six ®sh to be processed using one of four preparation methods. Fish were physically stunned using a heavy hammer, beheaded using a band saw (mod- * Author for correspondence. Tel: 765-496-3443; Fax: 765-494-0674; el 22, Biro Manufacturing Co., Marblehead, Ohio), then skinned E-mail: [email protected]. (Techo, Townsend Engineering Co., Des Moines, Iowa), and hand 764 SANTERRE ET AL. J. Food Prot., Vol. 63, No. 6

TABLE 1. Average chlordane residues in six raw and cooked ®sh ®llets from chlordane-fed cat®sh prepared by frying, baking, or smoking or analyzed raw Chlordane (ppb) mean concentration Ϯ SD (n ϭ 6)

Moisture content Lipid content Wet basis

Cooka Raw Cook % diff. Raw Cook % diff. Raw Cook

FR 76.9 Ϯ 1.6 60.5 Ϯ 3.2 Ϫ21.2 Ϯ 5.2 3.7 Ϯ 1.6 10.6 Ϯ 1.6 274 Ϯ 242 30 Ϯ 13 22 Ϯ 9 IF 75.8 Ϯ 1.2 67.6 Ϯ 1.5 Ϫ10.8 Ϯ 0.8 5.3 Ϯ 1.1 8.6 Ϯ 1.3 73 Ϯ 55 30 Ϯ 8 5 Ϯ 0 BK 75.1 Ϯ 2.5 61.9 Ϯ 4.1 Ϫ17.6 Ϯ 3.0 4.8 Ϯ 1.8 6.1 Ϯ 2.3 28 Ϯ 17 36 Ϯ 12 48 Ϯ 16 SM 75.6 Ϯ 1.3 65.6 Ϯ 1.8 Ϫ13.2 Ϯ 1.7 6.2 Ϯ 1.4 8.4 Ϯ 2.3 36 Ϯ 20 43 Ϯ 13 54 Ϯ 14 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/63/6/763/1673844/0362-028x-63_6_763.pdf by guest on 27 September 2021 a Cooking method: FR, deep fried; IF, polyphosphate injected and fried; BK, baked; SM, smoked. b Values in each column followed by different letters are signi®cantly different (P ϭ 0.05).

®lleted into two mirror-image ®llets keeping the belly ¯aps intact. Individual ®llets were ground twice using a meat grinder Matched ®llets were subsequently analyzed for the chlorinated (model 31FG10 Waring Products Div., New Hartford, Conn.). The residues with one ®llet being maintained raw while the other ®llet samples were placed in glass jars and stored at Ϫ23ЊC. Between was cooked using one of the four preparation methods. Corre- each sample, all utensils were cleaned thoroughly with detergent sponding ®llets from each ®sh were wrapped in aluminum foil (Alconox Inc., New York, N.Y.) rinsed with water, dried, and (Reynold Metals Co., Richmond, Va.) and placed in zip-lock plas- rinsed with 2-propanol (Baker Analyzed, J. T. Baker Inc., Phil- tic bags (Glad-Lock, First Brand Corp., Danbury, Conn.). The lipsburg, N.J.). bags were stored for 1 to 3 months at Ϫ23ЊC prior to further A 5-g ground sample of ®sh was weighed for moisture anal- processing at which time they were thawed at 4ЊC overnight. ysis. The sample was placed in an air oven at 105ЊC for 24 h and Frying (FR) of samples was done using a 10-liter deep fryer then weighed for weight loss. Extraction of fat and pesticide res- (Colinco Fish Cooker CC-10, Damark International, Minneapolis, idues from ®sh was performed using a modi®ed FDA method (9). Minn.) with canola oil (Crisco, Proctor & Gamble, Cincinnati, A 5-g sample was placed into a glass beaker and mixed with 75 Ohio). Breaded (House Autry, House Autry Mills Inc., Newton g of anhydrous sodium sulfate (Fisher Scienti®c), then placed in Grove, N.C.) ®llets were fried at 190ЊC for about 7 to 10 min a desiccator at room temperature for at least 12 h. The mixture until golden brown. An internal temperature of 71ЊC was obtained was transferred into a prerinsed 35-mm inner diameter by 90-mm and measured by a thermocouple. For a variation on the previous glass thimble (extra coarse, size 22) and extracted with a Soxhlet frying method, ®sh were injected (IF) with a 6% polyphosphate apparatus for at least 7 h using 300 ml hexane (Burdick and Jack- solution (Lem-O-fos, Rhone-Poulenc Basic Chemical Co., Shel- son, Muskegon, Mich.) at a turnover rate of between four and ®ve ton, Conn.) to add not more than 10% of ®llet weight. This is a times per hour. Included in each run of 10 samples were a matrix common commercial practice for maintaining quality following blank and a matrix spike. Recoveries for matrix spikes of between freezing and thawing. Samples were then stored at Ϫ23ЊC for 80 and 120% were acceptable. The extract was concentrated to about 4 weeks before they were breaded and fried as previously less than 5 ml by rotary evaporation at 40ЊC and transferred to a described. Samples were baked (BK) at 190ЊC in an oven (CPS preweighed, 16-mm ϫ 125-mm screw-top tube. The remaining 127, 27Љ single electric, Dacor, Pasadena, Calif.) for about 45 min solvent was evaporated and the residue weighed to determine fat until golden brown. Prior to smoking (SM), ®llets were soaked in content. a 25% salt solution (NaCl, SX0420-3, EM Science, Gibbstown, The extracted lipid was dissolved in 5 ml petroleum ether N.J.), at 10ЊC for about 1 h and air dried before smoking (140ЊC (distilled to a cut-off of 50ЊC, Phillips 66 Petroleum Ether 30-60, wet bulb, 160ЊC dry bulb for 1 h and 180ЊC wet bulb, 200ЊCdry Bartlesville, Okla.), and this solution was partitioned four times bulb for 1 h). Smoke was fed for the ®rst 30 min and total smok- with 30 ml acetonitrile (Burdick and Jackson) saturated with pe- ing time was about 2 h. All cooked ®llets were packaged as before troleum ether by shaking vigorously for 2 min each time. Each and stored at Ϫ23ЊC until further analysis. time the separated acetonitrile layer was transferred into a 1-liter

TABLE 2. Average toxaphene residues in six raw and cooked ®sh ®llets from toxaphene-fed cat®sh prepared by frying, baking, or smoking or analyzed raw Toxaphene (ppm) mean concentration Ϯ SD (n ϭ 6)

Moisture content Lipid content Wet basis

Cooka Raw Cook % diff. Raw Cook % diff. Raw Cook

FR 77.1 Ϯ 0.7 63.0 Ϯ 1.7 Ϫ18.3 Ϯ 2.3 4.9 Ϯ 0.9 10.8 Ϯ 1.2 125 Ϯ 51 0.160 Ϯ 0.071 0.126 Ϯ 0.045 IF 77.6 Ϯ 1.4 67.7 Ϯ 1.8 Ϫ12.7 Ϯ 3.1 4.6 Ϯ 1.5 8.0 Ϯ 1.1 88 Ϯ 50 0.145 Ϯ 0.048 0.125 Ϯ 0.052 BK 78.9 Ϯ 1.8 58.7 Ϯ 5.8 Ϫ25.5 Ϯ 7.8 4.9 Ϯ 2.3 6.3 Ϯ 2.2 36 Ϯ 25 0.133 Ϯ 0.041 0.167 Ϯ 0.058 SM 78.3 Ϯ 0.7 63.5 Ϯ 2.5 Ϫ19.0 Ϯ 2.9 4.2 Ϯ 0.6 7.7 Ϯ 1.7 83 Ϯ 36 0.122 Ϯ 0.021 0.150 Ϯ 0.056 a Cooking method: FR, deep fried; IF, polyphosphate injected and fried; BK, baked; SM, smoked. b Values in each column followed by different letters are signi®cantly different (P ϭ 0.05). J. Food Prot., Vol. 63, No. 6 CHLORDANE AND TOXAPHENE RESIDUES 765

TABLE 1. Extended

Chlordane (ppb) mean concentration Ϯ SD (n ϭ 6)

Wet basis Dry basis Fat basis

% diff.b Raw Cook % diff.b Raw Cook % diff.b

Ϫ25 Ϯ 20 F 131 Ϯ 48 56 Ϯ 26 Ϫ56 Ϯ 13 I 4,070 Ϯ 1,800 560 Ϯ 280 Ϫ84 Ϯ 9 R Ϫ82 Ϯ 5 G 123 Ϯ 31 15 Ϯ 1 Ϫ86 Ϯ 4 J 2,410 Ϯ 610 180 Ϯ 30 Ϫ92 Ϯ 1 R 33 Ϯ 8 H 142 Ϯ 39 124 Ϯ 34 Ϫ12 Ϯ 4 K 3,120 Ϯ 620 2,140 Ϯ 400 Ϫ30 Ϯ 9 S 29 Ϯ 36 H 179 Ϯ 55 157 Ϯ 39 Ϫ9 Ϯ 24 K 3,060 Ϯ 1,380 2,030 Ϯ 840 Ϫ33 Ϯ 12 S Downloaded from http://meridian.allenpress.com/jfp/article-pdf/63/6/763/1673844/0362-028x-63_6_763.pdf by guest on 27 September 2021

separatory funnel containing 650 ml deionized water, 40 ml sat- min, ramped at 10ЊC/min to 220ЊC, and held 20 min; total run urated NaCl solution (Fisher Scienti®c), and 100 ml petroleum time, 44 min. Standards were obtained from the Environmental ether. The separatory funnel containing the combined extracts was Protection Agency Repository (Research Triangle Park, N.C.). shaken vigorously for 2 min. Following phase separation, the The lower limits of quantitation for chlordane and toxaphene were aqueous layer was removed and extracted by shaking with 100 ml 0.01 and 0.05 ppm, respectively. petroleum ether. The aqueous layer was discarded and the petro- leum ether extracts combined and washed with two 100-ml por- RESULTS AND DISCUSSION tions of water containing 5 ml saturated NaCl. The petroleum ether extract was concentrated to less than 5 ml for further clean- Moisture content in raw ®llets averaged 77% with ®l- up. lets in the chlordane group averaging 76% and those in the The extract was transferred to a glass chromatographic col- toxaphene group averaging 78%. Variations can be attri- umn (22 mm inner diameter) containing 20 g of activated Florisil buted to differences in size, fat content, feed consumption, (PR grade, 60 to 100 mesh, U.S. Silica Co., Berkeley Springs, and ®lleting technique. Moisture decreased by an average W.V.). Florisil was activated at 130ЊC for 24 h and cooled to room of 17% to an average of 63.5% for cooked ®llets; however, temperature. The Florisil was topped with a 1-cm layer of anhy- there were variations observed between cooking methods drous sodium sulfate. The prepared column was rinsed with 50 and between as shown in Tables 1 and 2. ml of petroleum ether and the extract transferred onto the column. Lipid content, calculated on a dry basis (db), increased The column was eluted with 200 ml of 94% petroleum ether and 6% anhydrous diethyl ether (Fisher Scienti®c; containing 2% eth- following all cooking methods. The largest increase in fat anol, Quantum Chemicals Corp., Cincinnati, Ohio). The eluant occurred for FR ®llets for both chlordane (274% increase) was concentrated and exchanged into hexane for a ®nal volume and toxaphene (125% increase) samples. The range in lipid of 10 ml. An aliquot was transferred to 2 ml autosampler vials gain was from 28 to 274%. for analysis by gas chromatography. The changes in chlordane and toxaphene from raw ®l- Samples were analyzed using a Varian 3600 electron-capture lets to cooked ®llets are given in Tables 1 and 2. From a gas chromatograph ®tted with dual 0.53-mm inner diameter ϫ 30- wet basis calculation, it would appear that chlordane resi- ␮ m capillary columns (DB-5 [1.5 m ®lm thickness] and DB-608 dues increased when ®llets were baked or smoked. This can ␮ [0.83 m ®lm thickness]; J&W Scienti®c, Folsum, Calif.), a Var- be attributed to a reduction in moisture or a dehydration ian 8100 autosampler, and a DS-654 Data Station (quantifying effect that is only partially offset by an increase in lipid. A using peak area). Extract was injected into a single inlet that was split into the dual columns. Instrument settings were as follows: review of data (calculated on a wet basis) indicates that IF injector and detector temperatures, 230ЊC and 300ЊC, respectively; samples had a signi®cantly (P ϭ 0.05) greater reduction in hydrogen carrier gas; nitrogen make-up gas; temperature program- chlordane (Ϫ82%) than FR (Ϫ25%), BK (33%), or SM start at 150ЊC held 5 min, ramped at 5ЊC/min to 170ЊC, held 10 (29%). On a db, the same trend is observed. IF samples

TABLE 2. Extended

Toxaphene (ppm) mean concentration Ϯ SD (n ϭ 6)

Wet basis Dry basis Fat basis

% diff.b Raw Cook % diff.b Raw Cook % diff.b

Ϫ17 Ϯ 17 T 0.697 Ϯ 0.30 0.346 Ϯ 0.13 Ϫ49 Ϯ 9 W 14.4 Ϯ 5.5 3.3 Ϯ 1.6 Ϫ77 Ϯ 5 X Ϫ14.1 Ϯ 25 TU 0.639 Ϯ 0.17 0.392 Ϯ 0.18 Ϫ40 Ϯ 20 W 14.6 Ϯ 3.2 5.0 Ϯ 2.3 Ϫ65 Ϯ 18 XY 25 Ϯ 30 UV 0.631 Ϯ 0.19 0.399 Ϯ 0.12 Ϫ35 Ϯ 19 W 14.7 Ϯ 6.9 6.4 Ϯ 1.7 Ϫ51 Ϯ 15 Y 26 Ϯ 49 V 0.561 Ϯ 0.10 0.407 Ϯ 0.14 Ϫ24 Ϯ 31 W 13.4 Ϯ 2.1 5.3 Ϯ 1.7 Ϫ59 Ϯ 17 Y 766 SANTERRE ET AL. J. Food Prot., Vol. 63, No. 6 had an 86% chlordane reduction that was signi®cantly REFERENCES greater than FR (56%), BK (12%), or SM (9%). When com- 1. Akpan, V., M. Lodovici, and P. Dolara. 1994. Polycyclic aromatic pared on a percent fat basis (fb), the reduction in chlordane hydrocarbons in fresh and smoked ®sh samples from three Nigerian for IF (92%) and FR (84%) samples is not signi®cantly cities. Bull. Environ. Contam. Toxicol. 53:246±253. different; however, the reductions observed for BK (30%) 2. Armbruster, G., K. G. Gall, W. H. Gutenmann, and D. J. Lisk. 1989. and SM (33%) are signi®cantly lower than for fried ®llets. 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