f - 73-4* oil EPA ORDER NO. V/D(>99-06 ?3 >; JULY 1976 FINAL REPORT Region III Library Environmental Protection Agency P.CB.CON .,;-,,-A 13107 U.S. KNVfRONMtNTAL PROTIiCTION AGENCY RHGION III PHILAOHT.PHlA, PENNSYLVANIA 19106 EPA Report Collection Information Resource Center US EPA Region 3 Philadelphia, PA 19107 ------- EPA ORDER NO. WD6-99-0673-B JULY 1976 FINAL REPORT PCB CONCENTRATIONS IN STRIPED BASS AND EGGS , ,, ., COMPILED AND EDITED BY j.^ (/V ,} f.'1'- ,,,:.;S:v£Ct JOSEPH M. FORNS f.--.-..-,3 PA 19107 WESTINGHOUSE OCEAN RESEARCH LABORATORY ANNAPOLIS, MARYLAND SUBMITTED TO THOMAS O. MUNSON, PhD. PROJECT OFFICER OFFICE OF THE CHESAPEAKE BAY PROGRAM U.S. ENVIRONMENTAL PROTECTION AGENCY <-- REGION III PHILADELPHIA, PENNSYLVANIA 10106 APPROVED BY: ------- T TABLE OF CONTENTS I. INTRODUCTION " 1 II. METHODS AND MATERIALS 2 A. Collection and Preparation of Samples for Chemical Analysis 2 3. Chemical Analysis 4 III. RESULTS 5 IV. BIBLIOGRAPHY 10 ------- I. Introduction Polychlorinated biphenyls (PCB's) have been increasingly recognized as a significant environmental pollutant. In a previous survey of the Upper Chesapeake Bay conducted by Westinghouse Ocean Research Laboratory under contract to the Maryland Department of Natural Resources, the PCB's were found- to average 12 ppt in the water column - nearly the same value found in Lake Michigan where the salmonoid fish species are unfit for human consumption (exceeding FDA tolerance levels of 5 ppm) (Munson, et. al., 1975). Laboratory experiments have shown that some fish species accumulate PCB's more than 10,000 times those levels present in the water. .Therefore, PCB's in the water column at even parts per trillion levels could have significant detrimental effects on the fishery resource (Kleinert, 1975). Many experiments with finfishes have indicated that exposure to PCB's has in many cases hindered or prevented reproduction (Hansen, 1975; Nebeker, 1975), A PCS concentration as low as 1 ppm has been shown to drastically reduce the hatachability of trout eggs. PCB accumulation in the water column and consequently in fishes might therefore have an additional less immediately evident but possibly more detrimental and lasting effect on the fishery. In 1975, an estimated total of 8,604,000 pounds of striped bass were caught in the U.S. and sold at ar. ex-vessel price to the fishermen of $3,993,000.00. Of those, 2,722,216 pounds were caught in Maryland, at a value of $1,064,168.00 (NXFS, 1976). It was ir. light of these facts that this study was undertaken. Its aims are twofold. First, the results serve as a good approximation of PCB levels in the: striped bass which spawn in the bay. This gives an indication of any immediate danger to the striped bass fishery which might exist in the form of unacceptable PCB concentrations in the fish flesh. In addition, in the analysis of striped bass roe for PCB content, this study serves as a pilot study for future ones which might possibly test the effects of PCB's on striped bass reproduction. ------- II. METHODS AND MATERIALS A. Collection and Preparation of Samples for Chemical Analysis Thirty-three female, two male and two striped bass of unknown sex were collected from the following spawning areas in the Chesapeake Bay region (Figure 1): The Nanticoke, Choptank, Bohemia, Sassafras, Elk, Little Elk, Potomac, Rappahannock, and James Rivers. Due to an unusually early spawn and a delay in the contract award, no samples were obtained from the Patuxent River. Because-the James River was closed to fishing during- the spawning period, no samples were collected there during that time. However, previously collected and frozen samples of female fish without roe were obtained for the study through Dr. Robert Huggett at the Virginia Institute of Marine Sciences. Assisting in providing samples for the program were the Chesapeake Bay Institute, the Chesapeake Biological Laboratory, the Fisheries Department of the Maryland Department of Natural Resources, and the Annapolis Field Office of the Environmental Protection Agency (Region 3). In the lab, the length to the nearest centimeter, the weight to the nearest gram, and the general condition of each fish (excluding the VIMS samples) was recorded. The sex of each individual was determined as well. Each fish wss scaled and cleaned. The liver from each fish was frozen. From each feiaale fish, one ovary was removed and frozen whole. From the remaining ovary several subsamples of roe ware weighed out and frozen individually. A one to one and one half inch steak as taken from each fish just posterior to the pectoral fins and anterior to the dorsal fin, wrapped in aluminum foil and frozen immediately. The remainder of each fish was wrapped and frozen whole and is being stored at the Westinghouse Annapolis facility awaiting transfer to the Annapolis Field Office of EPA. The frozen meat and roe subsamples were prepared for chlorinated hydrocarbon analysis at the Annapolis facility. Meat subsamples were thawed, diced and blended to a. purSe. Approximately twentyfive gram aliquots of meat subsamples were weighed to the nearest hundredth gram. Roe samples were thawed and 25 gram aliquots of eggs from each fish for which eggs were available were also weighed out. Each of these meat and egg samples was then Individually ground in a mortar and pestle with three to five times its wet weight in anhydrous sodium sulfate. These samples were allowed to stand for about thirty minutes and then ground again to insure against caking. The powdered preparations were then double wrapped in aluminum foil, taped, labeled and keot dry at room temperature under derfsicnnt prior Co sh Lpm-at for atuiiy.sis. AIL T.iat^r^.al.^,, \ .-.'.,, glassware, blender parts, mortars and pestles, were washed in hoc soapy water, rinsed thoroughly, dried and subsequently rinsed in acetone and hexane prior to use in the preparation of each sample. ------- 77"!00' 76°iOO' 75°lOO' ^4J^-^? ^.-,--,:..,,. --.. BALTIMORE ^/V7-*-£~S^' ^T-A \^M;' -\'.':-:--.:---: [TT^V- r\ \^->vm- ^t;^ f^lr^' *r - .-:-=.7Vi i% ---,,v^ :./..^>%|fv;;-.^V-| -:.::-;.; -.T*--.-.-. - -.-A :., .'." :-.--.:.-" $*' - - $:: \ 1 * - v -A-^'i . cv - r --^" ^^-V i- -^ i . 4 ^'^ ^^ -U4 V ^ V-v/ - O t ---.St" if ^NORFOLK ^ _;,. -';;v- vy-^X%^g,-,:^ r^ltC; ?! 5° DO' FIGURE 1. Spawning areas of the Chesapeake Bay region used as sampling locations. ------- B. Chemical Analysis The gas chromatographic analysis of the samples prepared as described above were- performed by the Analytical Bio Chemistry Laboratories (P. 0. Box 1097/ Columbia, Missouri 65201). For each sample, the sample matrix was packed in a 22 mm inside diameter (ID) column and extracted by percolating 250 milliliters of 6% diethyl ether hexane through the column. The ^ eluate was collected, evaporated, and brought to a 25 milliliter volume. The sample weights were provide'd by Westinghouse and the sample concentration was generally 1 gram per milliliter. A 5 milliliter aliquot was drawn and added to a 4 inch florisil column topped with 1/2 inch of anhydrous sodium sulfate. The polychlorinated biphenyls were eluded with 150 milliliters of hexane. The eluate was concentrated to 10 milliliters and a few microliters were injected with a Ni,_ electron capture detector. The GLC conditions were as follows: Injector: 235°C Column: 2Q5°C a 1.5% OB17 2.0% OV210 on Gas Chromatograph Q 100/120 mesh, 4 nun ID by 6' length, l^ flow approximately 100 ml/minute Detector: 3CO°C Nig-3 electron capture detector with .. n an electrometer setting 3 x 10 amp full scale. A 2 milliliter aliquot of the diethyl ether hexane extract was drawn and evaporated and the residue weighed and reported as % fat. All calculations were based on the extract sample weights supplied by Westinghouse. Quantisation and identification of PCBs pose sever?.! problems, especially when they exist in a weathered condition. Consequently, the last four peaks of ten peaks for Aroclor 1254 were summed and used for quantitation. This was to avoid p,p'-DDE and chlordane interferences. The last eight peaks of eleven peaks were summed and used for quantitation of Aroclor 1260. The first five peaks of eleven total peaks of Aroclor 1242 were summed and used for quantitation. These were judgements based on peak height ratios and possible interferences which might exist in the sample extract. For the extracts which have p,p'-DDE and chlordane components in tae .ieaUj f.h-: PCB^s will be s-ep-iratsd from chlordane and >, o '-DOR "ovpnnentr; 03; silicic acid column separations. The^e additional samples cleaned up by silicic acid will verify the quantitative results for the PCBs obtained in the analyses described above and will also give semi-quantitative results for chlordane and p,p'-DDE. ------- III. RESULTS The ppm concentrations of the PCBs found in the samples are given in Table I, as well as the % .fat in each sample. The average PCS concentrations in the fish meat samples and the average concentrations in the egg samples from each individual spawning area surveyed are shown on the map in Figure 2. Figure 3 graphically describes the results of the linear regression analysis comparing ppm PCB concentrations in the flesh of each female fish to that in her eggs. Figure 4 shows the results of the same analysis after the ppm concentrations have been corrected taking into consideration the different % fat values in each of the samples. Copies of the Gas Chromatograph traces are included in Appendix A of this report, ------- TABLE I PCS CONTENT IN FEMALE AND EGGS OF STRIPED BASS FROM THE CHESAPEAKE BAY Sample Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 I"7 18 19 20 21 22 23 24 25 26 27 *28 *29 *30 *31 *32 *33 *34 *37 PCB Content in ppm Location Nanticoke Nanticoke Nanticoke Nanticoke Nanticoke Potomac Potomac Potomac Potomac Potomac Sassafras Sassafras Bohemia Bohemia Bohemia Bohemia Bohemia L. Elk L. Elk Elk Elk Elk Choptank Choptank Choptank Choptank Choptank James James James James James Rappahannock Rappahannock Hap p aha r.no c k Rappahannock Rappahannock Length (cm.) 61.60 97.16 91.44 68.58- 67.31 65.53 72.39 65.04 64.77 60.96 60.50 62.30 63.30 54.7 57 .0 59,3 68.5 69.5 59.4 61.4 58.0 51.0 80.01 76.84 80.01 59.06 -60.96 93.0 59.0 74.0 69.0 74.0 76.0 67.0 63.0 59 ,0 81.0 Weight (gm.) 3,899 13,381 9,072 4,990 3,856 3,289 5,239 5,353 4,082 3,969 3,740 4,086 3,987 2,723 2,910 3,680 5,584 5,770 3,528 3,935 3,219 3,224 7,938 7,371 7,031 3,629 3,515 10,860 3,040 6,460 5,660 5,430 6,865 5,085 4,095 6, 545 7,980 ppm 0.46 0.48 0.47 0.47 0.54 0.47 0.42 0.14 0.50 0.24 0.15 0.11 0.26 0.27 0.20 ,0.08 10.14 0.51 0.24 0.07 <0.06 0.12 0.05 0.24 0.21 0.11 0.14 0.08 1.27 0.62 0.67 1.32 1.65 10 Q j- j,\j « y * 1 f\ O ID. y 1.14 1,43 0,o2 1.66 Flesh PCB 1260 1254 1254 1254 - 1260 1254, 1254 1260 1254 1254 1254 1254 1260 1254 1254 1242, 1260J 1254 1254 1254 1254 1254 1254 1260 1260 1254 1254 1260 1260 1260 1260 1260 1260 1742 JL ifc.*-T J -i 1 O C/, f -L/34- 1260 1260 1250 1260 % fat 2.8 0.8 0.3 2.5 2.3 2.0 1.8 1.3 1.4 1.4 1.3 0.8 1.2 1.5 1.1 0.9 2.5 1.8 0.7 1.0 0.9 1.0 0.7 0.6 0.5 0.9 0.5 4.3 3.2 5.9 7.1 7.7 10.3 7.8 10.5 10.9 9.7 Ppm 2.56 5.75 11.2 3.08 (male) 3.54 2.63 3.08 6.57 1.68 2.02 3.72 4.17 4.32 3.07 3.88 4.44 3.16 2.46 1.32 4.36 1.77 7.12 6.47 4.00 2.74 3.04 Eggs PCB 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 1260 % fat 29.6 19.7 22.6 33.3 31 ..2 32.6 28.9 24.2 14.2 24.5 26.7 28.2 29.1 28.5 27.1 28.6 32.2 29.6 31.3 35.4 30.8 28.7 29.4 30.3 14.1 21.9 * Female meat only from VTMS ------- 7 7 "TOO1 76'JOO' 750J001 391 00' ' .is WASHING" ;-';.:.::2.87^ I --- _i. y*» s. ""-- **- * -i?-^- j ii~>.3 ,K-;~|||pg|||p ---::^;>^^-Y--^o-s*>---i> x &?v - \?^-::"'^S^-^:^i^^l ^^^^^te^^^^^^d """""'"" ^?""S:^-v^^^l. .16 ^ %--,?.; :.--'-v--:v^l;s^:5 ;-;;v;v V^:\r"4.67 ,t%^V'-:C ^"'. -^."-':.:-"-:^":??S^7rV MI '-- ".-----:----.:-.:.."=--^-^-""v?=. .-*!,;" -^ V^£ ' H] ---:-..-:. -."-.:: .:.-;.:..- ^-/4:_ --^S ; .. ^^ '-/ = |li :-- I'- -----------.V ^V:"-*^/.-.--:-X" -' " - if, ' " r - ^"^V: -'',> j \.->f-_v_ 'oo'jl1 D^__ ,r 77 OO' 76"IOO' 75" (00* FIGURE 2. Average concentrations (ppm) o£ PCBs in Striped Bass flesh and eggs from each of the sampling areas. (* no eggs available for study) ------- 12.0 -r 0100 0.200 0.300 PCB IN FLESH (PPM) 0.400 0500 76128A01 Figure 3. Relationship ot PCB Concentration in Female Striped Bass Flesh and Their Fggs. ------- 2.0 V) o CD 111 u. O < Li. 6? CO CJ OL 1-8 J. 1.6 _L 1.4 J. 1.2 1.0 0.8 0.6 0.4 0.2 r = 0.0185 JL J 0.005 PCB IN % FAT OF FLESH (PPM) 0.010 Figure 4. Relationship of PCB Concentration in Fat Content of Female Striped Bass Flesh E 9 76128A02 and Their Eggs. ------- BIBLIOGRAPHY Hansen, David J., "PCB's: Effects on and Accumulation by Estuarine Organisms," -. - ^National Conference on Polychlorinated Biphenyls; Conference Proceedings, Environmental Protection Agency Office of Toxic Substances, Washington, B.C. (March, 1976), EPA-560/6-75-004. pp. 282-283.' Kleinert, Stanton., "Sources of Polychlorinated Biphenyls in Wisconsin," National Conference on Polychlorinated Biphenyls; Conference Proceedings, Environmental Protection Agency Office of Toxic Substances, Washington, D.C. (March, 1976), EPA- 560/6-75-004. pp. 124-126. Munson, T.O., Palmer, H.P., and Forns, J.M., "Transport of Chlorinated Hydro- carbons in the Upper Chesapeake Bay," National Conference on Polychlorinated Biphenyls: Conference Proceedings', -Environmental Protection Agency Office of Toxic Substances,, Washington, D.C. (March, 1976), EPA560/6-75-004. pp. 218-219. National Marine Fisheries Service, Fisheries of the U.S., 1975, Current Fisheries Statistics No. 6900 (March, 1976). Nekeker, Alan, V., "Summary of Recent Information Regarding Effects of PCB's on Freshwater Organisms." National Conference on Polychlorinated Biphenyls: Conference Proceedings, Environmental Protection Agency Office of Toxic Substances, Washington, D.C. (March, 1976), EPA560/6-75-004, pp. 284-291. 10 ------- EPA ORDER NO. WD6-99-0673-B JULY 1976 APPENDIX A TO PCB CONCENTRATIONS IN STRIPED BASS AND EGGS i COMPILED AND EDITED BY f ff' JOSEPH M. FORNS J WESTINGHOUSE OCEAN RESEARCH LABORATORY | ANNAPOLIS, MARYLAND I SUBMITTED TO THOMAS O. MUNSON, PhD. PROJECT OFFICER OFFICE OF THE CHESAPEAKE BAY PROGRAM U.S. ENVIRONMENTAL PROTECTION AGENCY REGION III PHILADELPHIA, PENNSYLVANIA 19106 ------- |