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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
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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:
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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
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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.
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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.
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sampling locations.
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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.
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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,
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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
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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
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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
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FIGURE 2. Average concentrations (ppm) o£ PCBs in Striped Bass
flesh and eggs from each of the sampling areas.
(* no eggs available for study)
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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.
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0.8
0.6
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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.
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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
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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
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